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PROTECTED DISACCHARIDES, THEIR PROCESS OF PREPARATION AND THEIR USE IN THE SYNTHESIS OF ZWITTERIONIC OLIGOSACCHARIDES, AND CONJUGATES THEREOF

20240024489 ยท 2024-01-25

    Inventors

    Cpc classification

    International classification

    Abstract

    The present invention provides zwitterionic oligosaccharides, in particular fragments of the surface polysaccharides from Shigella sonnei and Shigella sonnei conjugates comprising them. The present invention also provides protected disaccharides, their process of preparation and their use in the synthesis of zwitterionic oligosaccharides, and conjugates thereof, the disaccharide repeating unit of Shigella sonnei being: (I)

    ##STR00001##

    Claims

    1. A conjugate comprising an oligo- or polysaccharide selected from the group consisting of:
    (B).sub.x-(A-B).sub.n-(A).sub.y, and
    (A).sub.x-(B-A).sub.n-(B).sub.y, wherein: x is 0 or 1, y is 0 or 1, n ranges from 1 to 50, in particular from 1 or 2 to 10, more particularly from 1 or 2 to 4 or from 3 to 8, n being notably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, A is 4)--L-AltpNAcA-(1.fwdarw., B is 3)--D-FucpNAc4N-(1.fwdarw., or a pharmaceutically acceptable salt thereof, said oligo- or polysaccharide being bound to a carrier, in particular covalently bound to a carrier.

    2. The conjugate according to claim 1, wherein the carrier is selected among a protein or a peptide comprising at least one T-helper epitope, or a derivative thereof.

    3. The conjugate according to claim 1, wherein the oligo- or polysaccharide is bound to the carrier via a spacer which does not contain any carbohydrate residue.

    4. The conjugate according to any one of the claims 1 to 3, characterised in that the ratio of oligo- or polysaccharide to the carrier ranges from 1:1 to 500:1, in particular between 1:1 and 200:1, more particularly, between 1:1 and 30:1, preferably between 5:1 and 25:1, more preferably between 8:1 and 30:1, 40:1 or 50:1, or between 5:1 and 20:1.

    5. An immunogenic composition comprising a conjugate according to any one of claims 1 to 4 and a physiologically acceptable vehicle.

    6. The immunogenic composition according to claim 5, characterised in that it is formulated for parenteral, oral, intranasal or intradermal administration.

    7. The conjugate according to any one of claims 1 to 4 or the immunogenic composition according to claim 5 or 6, for use in vaccination, in particular against S. sonnei infection and/or infection caused by pathogens featuring cross-reactive carbohydrate antigens, for example a Plesiomonas shigelloides infection, notably a P. shigelloides O17 infection.

    8. A compound of the following formula:
    Q-(B).sub.x-(AB).sub.n-(A).sub.y-OR(IIa) or
    Q-(A).sub.x-(BA).sub.n-(B).sub.yOR(IIb), wherein: x is 0 or 1, y is 0 or 1, n ranges from 1 to 50, Q is H or a C.sub.1-C.sub.6 alkyl, A is 4)--L-AltpNAcA-(1.fwdarw., B is 3)--D-FucpNAc4N-(1.fwdarw., R is H, C.sub.1-C.sub.6 alkyl, in particular propyl or methyl, or a group LZ, L is: a single bond, a divalent C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl or C.sub.2-C.sub.12 alkynyl chain optionally interrupted by one or more heteroatoms, notably selected from an oxygen atom, a sulphur atom or a nitrogen atom, said nitrogen and sulphur atoms being optionally oxidized, and the nitrogen atom being optionally involved in an acetamide bond, or a divalent C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl or C.sub.2-C.sub.12 alkynyl chain substituted by at least one OH group, being in particular of the following formula (CH.sub.2CH.sub.2C(OH)).sub.q(CH.sub.2CH.sub.2).sub.i, wherein i is 0 or 1 and q ranges from 1 to 10, N(R.sup.a)-D-, wherein R.sup.a is H, C.sub.1-C.sub.4-alkyl, C.sub.1-C.sub.4-alkoxy, CH.sub.2C.sub.6H.sub.5, CH.sub.2CH.sub.2C.sub.6H.sub.5, OCH.sub.2C.sub.6H.sub.5, or OCH.sub.2CH.sub.2C.sub.6H.sub.5; D is C.sub.1-C.sub.7-alkylene, C.sub.1-C.sub.7-alkoxy, C.sub.1-C.sub.4-alkyl-(OCH.sub.2CH.sub.2).sub.pOC.sub.1-C.sub.4-alkyl, OC.sub.1-C.sub.4-alkyl-(OCH.sub.2CH.sub.2).sub.pOC.sub.1-C.sub.4-alkyl or C.sub.1-C.sub.7-alkoxy-R.sup.b, wherein R.sup.b is an optionally substituted aryl or an optionally substituted heteroaryl, and wherein p is 0 to 6, preferably p is 1, 2 or 3, and further preferably p is 1; Z is Z.sub.1 or F.sub.1-L.sub.2-Z.sub.2, Z.sub.1 is a terminal function or group, optionally protected, able to form a covalent bond with a carrier and/or a solid support, or a multivalent scaffold; an anchor; a mono-, oligo- or polysaccharide; or a dye or fluorescent residue. F.sub.1 is any group enabling to bond the linker L to the linker L.sub.2, F.sub.1 being in particular chosen from the C(O), C(O)C(O), C(O)C(O)NH, NHC(O)C(O), NHC(O)C(O)NH, C(O)C(H)NNH, NHC(O)C(H)NNH, ester, amide, amine, CH.sub.2, ether, thioether, imine, thio-succinimide, oxime, hydrazone, hydrazonamide, C(O)CH.sub.2NH, NHCH.sub.2C(O), triazole functions or groups, and from the following: ##STR00127## F.sub.1 being more particularly chosen from thio-succinimide, C(O)CH.sub.2NH, NHCH.sub.2C(O), and triazole functions or groups, L.sub.2 is a single bond, divalent C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl or C.sub.2-C.sub.12 alkynyl chain optionally interrupted by one or more heteroatoms, notably selected from an oxygen atom, a sulphur atom or a nitrogen atom, said nitrogen and sulphur atoms being optionally oxidized, and the nitrogen atom being optionally involved in an acetamide bond, Z.sub.2 is Z.sub.1 or F.sub.2-L.sub.3-Z.sub.1, F.sub.2 is any group enabling to bond the linker L to the linker L.sub.3, F.sub.2 being in particular chosen from the C(O), C(O)C(O), C(O)C(O)NH, NHC(O)C(O), NHC(O)C(O)NH, C(O)C(H)NNH, NHC(O)C(H)NNH, ester, amide, amine, CH.sub.2, ether, thioether, imine, thio-succinimide, oxime, hydrazone, hydrazonamide, C(O)CH.sub.2NH, NHCH.sub.2C(O), triazole functions or groups, and from the following: ##STR00128## F.sub.2 being more particularly chosen from thio-succinimide, C(O)CH.sub.2NH, NHCH.sub.2C(O), and triazole functions or groups, L.sub.3 is single bond, a divalent C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl or C.sub.2-C.sub.12 alkynyl chain optionally interrupted by one or more heteroatoms, notably selected from an oxygen atom, a sulphur atom or a nitrogen atom, said nitrogen and sulphur atoms being optionally oxidized, and the nitrogen atom being optionally involved in an acetamide bond, with the proviso that said compound is not H-ABOPr, HBA-OPr, H-ABA-OPr, HBABOPr, H-(AB).sub.2OPr or H-BA-OMe, or a pharmaceutically acceptable salt thereof.

    9. The compound according to claim 7, selected from the group consisting of: ##STR00129## with in particular n=1, 2 or 3, ##STR00130## with in particular n=1, 2, 3 or 4, ##STR00131## with in particular n=1, 2, 3, 4 or 5, ##STR00132## with in particular n=1, 2, 3 or 4, ##STR00133## with in particular n=1, 2, 3 or 4, ##STR00134## with in particular n=1, 2 or 3, ##STR00135##

    10. Kit for the in vitro diagnostic of S. sonnei infection, wherein said kit comprises a compound according to claim 8 or 9, optionally bound to a label or a solid support.

    11. Use of a compound of the following formula (I.sub.0):
    T-A-BY or T-B-A-Y(I.sub.0), wherein: T is chosen from 2-naphtylmethyl (Nap), para-methoxybenzyl (PMB), 4-bromobenzyl (PBB), benzyloxymethyl acetal (BOM), 2-methoxyethoxymethylether (MEM), methoxypropyl (MOP), tetrahydropyranyl (THP), allyl (All), C.sub.1-C.sub.6 alkyl, or a silyl, T being in particular tert-butyldimethylsilyl (TBS), dimethylhexylsilyl (TDS), triisopropyl silyl (TIPS), or triethylsilyl (TES), Y is chosen from: OAll, when T is not All; Silyl ethers, in particular tert-butyldimethylsilyl ether (OTBS), dimethylhexylsilyl ether (OTDS), triethylsilyl ether (OTES), triisopropyl silyl ether (OTIPS), when T is Nap or PMB; OPMB, ONap, when OT is a silyl ether or T is All or PBB ether; p-methoxyphenyl-O (OMP or OPMP); and SR.sub.4, wherein R.sub.4 is such as the compound is a thioglycoside; A is ##STR00136## in particular ##STR00137## wherein: P.sub.1 is chosen from TCA, TFA, DCA, CA, Ac, benzyloxycarbamate (Cbz), Trichloroethoxycarbonyl (Troc), and Fmoc, at least one P.sub.1 and P.sub.2 being chosen from TCA, DCA, Ac, Fmoc, Troc, and, when Y is not OAll, OAlloc, P.sub.2 is H or chosen from Ac, Boc, TFA, benzyloxycarbamate (Cbz), and 2,2,2-trichloroethoxycarbonyl (Troc), P.sub.2 being H when P.sub.1 is not Ac, or P.sub.1 and P.sub.2 form together a phthalimido or a tetrachlorophthalimido (Cl.sub.4Phth) group, R.sub.2 is CO.sub.2R.sub.1 or CH.sub.2OR.sub.3, wherein R.sub.3 is Ac, benzoyl (Bz), or R.sub.3 forms together with group T a benzylidene group, R.sub.1 is chosen from C.sub.1-C.sub.6 alkyl, notably Me or tert-butyl (tBu), Bn and p-methoxybenzyl (PMB) groups, R.sub.1 being in particular Bn, B is ##STR00138## in particular ##STR00139## for the preparation of a compound of the following formula (II) Q-(B).sub.x-(AB).sub.n-(A).sub.y-OR (IIa) or Q-(A).sub.x-(BA).sub.n-(B).sub.yOR (IIb) according to claim 8 or 9.

    12. Process of preparation of a compound of the following formula (II): Q-(B).sub.x-(AB).sub.n-(A).sub.y-OR (IIa) or Q-(A).sub.x-(BA).sub.n-(B).sub.yOR (IIb), according to claim 8 or 9 said process comprising the following steps: (i) a step of converting a compound of following formula T-A-BY, in particular T-A-BOAll, or T-B-A-Y (I.sub.0) as defined in claim 11 into a donor compound of following formula T-A-BX or T-B-A-X (I.sub.D), by intermediately forming the hemiacetal T-A-BOH or T-B-A-OH, wherein X represents a leaving group chosen from imidates, for example OPTFA or OTCA, PTFA representing N-phenyltrifluoroacetimidyl and TCA representing trichloroacetimidyl, from o-alkynylbenzoates, and from diphenyl oxosulfoniums, in particular, when Y is All, through metallo-catalyzed deallylation, for example Pd, Ir or Rh, more particularly in presence of H.sub.2-activated Ir-catalyst, then aqueous I.sub.2 or NIS, with optionally a base, in particular an inorganic base, for example NaHCO.sub.3, or by Pd-catalyzed deallylation, in particular in presence of PdCl.sub.2, then aqueous I.sub.2 or NIS, with optionally a base, in particular an inorganic base, for example NaHCO.sub.3, to provide the corresponding hemiacetal T-A-BOH or T-B-A-OH and then PTFA-Cl or trichloroacetonitrile, and/or (ii) a step of converting a compound of following formula T-A-BY, in particular T-A-BOAll, or T-B-A-Y (I.sub.0) with T being not C.sub.1-C.sub.6 alkyl, into an acceptor compound of following formula H-A-BY, in particular H-A-BOAll, or HB-A-Y (I.sub.A), in particular in presence of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), CAN or an acid when T is Nap or PMB, or in presence of buffered TBAF, for example buffered with AcOH, or Et.sub.3N.Math.3HF, when T is a silyl, or in presence of an organic, inorganic, or Lewis acid, such as AcOH, TsOH, HCl, ZnBr.sub.2 when T is THP, MEM, MOP, and/or (iii) a step of obtaining from compound (I.sub.A) and/or (I.sub.D) a compound Q-(B).sub.x-(A-B).sub.m (A).sub.y-Y, in particular Q-(B).sub.x-(A-B).sub.m-(A).sub.y-OAll, or Q-(A).sub.x-(B-A).sub.m-(B).sub.yY (II.sub.OP), with m being from 1 to n, and Q being T when x is 0 and chosen from T, Bn and acyl groups, for example Lev, ClAc, Fmoc, or Ac when x is 1, in particular in presence of a Lewis acid, for example chosen from TMSOTf, TBSOTf, TfOH, Yb(OTf).sub.3, Cu(OTf).sub.2, AgOTf, or boron trifluoride etherate, (iv) when R is LZ and L is not N(R.sup.a)-D-, a step of conjugating compound (II.sub.OP) into a compound of following formula Q-(B).sub.x-(A-B).sub.m-(A).sub.y-OLZ or Q-(A).sub.x-(B-A).sub.m-(B).sub.y-OLZ (II.sub.CP), or a compound of following formula Q-(B).sub.x-(A-B).sub.m-(A).sub.y-OW or Q-(A).sub.x-(B-A).sub.m-(B).sub.yOW, wherein W is L-F.sub.1 or L-F.sub.1.sub.P, L being as defined above, F.sub.1 being a precursor of F.sub.1 as defined above, F.sub.1.sub.P being a protected group F.sub.1, in particular with one or more benzyl groups, or when R is LZ and L is N(R.sup.a)-D-, a step of preparation of a compound Q-(B).sub.x-(A-B).sub.m (A).sub.y-OH or Q-(A).sub.x-(B-A).sub.m-(B).sub.yOH, (iv) optionally, when m is not n, a step of converting a compound Q-(B).sub.x-(A-B).sub.m-(A).sub.y OLZ or Q-(A).sub.x-(B-A).sub.m-(B).sub.y-OLZ (II.sub.CP), or a compound of following formula Q-(B).sub.x-(A-B).sub.m-(A).sub.y-OW or Q-(A).sub.x-(B-A).sub.m-(B).sub.yOW to a compound Q-(B).sub.x-(A-B).sub.n-(A).sub.y-OLZ or Q-(A).sub.x-(B-A).sub.n-(B).sub.y-OLZ (II.sub.CP), or a compound of following formula Q-(B).sub.x-(A-B).sub.n-(A).sub.y-OW or Q-(A).sub.x-(B-A).sub.n-(B).sub.yOW respectively, being noted that when the Q group of Q-(B).sub.x-(A-B).sub.m-(A).sub.y-OLZ or Q-(A).sub.x-(B-A).sub.m-(B).sub.y-OLZ (II.sub.CP) or Q-(B).sub.x-(A-B).sub.m-(A).sub.y-OW or Q-(A).sub.x-(B-A).sub.m-(B).sub.yOW is not C.sub.1-C.sub.6 alkyl, the Q group of Q-(B).sub.x-(A-B).sub.n-(A).sub.y-OLZ or Q-(A).sub.x-(B-A).sub.n-(B).sub.y-OLZ (II.sub.CP) or Q-(B).sub.x-(A-B).sub.n-(A).sub.y-OW or Q-(A).sub.x-(B-A).sub.n-(B).sub.yOW can represent C.sub.1-C.sub.6 alkyl, (v) a step of deprotection of the compound obtained after step (iii) or (iv) to obtain the compound of following formula Q-(B).sub.x-(AB).sub.n-(A).sub.y-OLZ or Q-(A).sub.x-(BA).sub.n-(B).sub.yOLZ (II) or a compound of following formula Q-(B).sub.x-(AB).sub.n-(A).sub.y-O-L-F.sub.1 or Q-(A).sub.x-(BA).sub.n-(B).sub.yO-L-F.sub.1, or a compound of following formula Q-(B).sub.x-(AB).sub.n-(A).sub.y-OH or Q-(A).sub.x-(BA).sub.n-(B).sub.yOH, in particular in presence of Pd(OH).sub.2C or PdC, H.sub.2, for example generated as high-pressure hydrogen with the electrolysis of water, and a base, in particular an inorganic base, for example chosen from NaHCO.sub.3, K.sub.2CO.sub.3, NH.sub.4HCO.sub.3, CaCO.sub.3, MgCO.sub.3, and optionally followed by saponification then in presence of organic/inorganic base for example ethylenediamine, triethylamine, diethylamine, hydoxylamine, NH.sub.2OH or of LiOH/H.sub.2O.sub.2, when R.sub.1 is C.sub.1-C.sub.6 alkyl, notably Me, or before in presence of TBAF or TFA, ZnBr.sub.2, TsOH, when T is a silyl ether, THP, MEM, MOP and/or P.sub.1 is Boc, (vi) when the compound obtained in step (v) is of formula Q-(B).sub.x-(AB).sub.n-(A).sub.y-O-L-F.sub.1 or Q-(A).sub.x-(BA).sub.n-(B).sub.yO-L-F.sub.1, a step of contacting said compound with a compound of following formula F.sub.1-L.sub.2-Z.sub.1, F.sub.1 being a precursor of F.sub.1 as defined above, L.sub.2 and Z.sub.1 being as defined above, or a compound of following formula F.sub.1-L.sub.2-F.sub.2, F.sub.1 being a precursor of F.sub.1 as defined above, F.sub.2 being a precursor of F.sub.2 as defined above, L.sub.2 being as defined above, followed by contacting the obtained compound with a compound of following formula F.sub.2-L.sub.3-Z.sub.1, wherein F.sub.2 is a precursor of F.sub.2 as defined above, and L.sub.3 and Z.sub.1 being as defined above, or when the compound obtained in step (v) is of formula Q-(B).sub.x-(AB).sub.n-(A).sub.y-OH or Q-(A).sub.x-(BA).sub.n-(B).sub.yOH, a step of contacting said compound with: a compound of following formula HN(R.sup.a)-D-Z.sub.1, R.sup.a, D and Z.sub.1 being as defined above, or a compound of following formula HN(R.sup.a)-D-F.sub.1, F.sub.1 being a precursor of F.sub.1 as defined above, followed by contacting the obtained compound with a compound of following formula F.sub.1-L.sub.2-Z.sub.1, wherein F.sub.1 is a precursor of F.sub.1 as defined above, to give the compound of following formula Q-(B).sub.x-(AB).sub.n-(A).sub.y-OLZ or Q-(A).sub.x-(BA).sub.n-(B).sub.y-OLZ (II).

    13. Compound of one of the following formulae (III):
    Q-(B).sub.x-(A-B).sub.m-(A).sub.y-Y or Q-(B).sub.x-(A-B).sub.m-(A).sub.y-OLZ,Q-(B).sub.x-(A-B).sub.m-(A).sub.y-O-L-F.sub.1,Q-(B).sub.x-(A-B).sub.m-(A).sub.y-O-L-F.sub.1.sub.P,Q-(B).sub.x-(A-B).sub.m-(A).sub.y-OH,H(B).sub.x-(A-B).sub.m-(A).sub.yY,Q-(A).sub.x-(B-A).sub.m-(B).sub.yY or Q-(A).sub.x-(B-A).sub.m-(B).sub.y-OLZ,Q-(A).sub.x-(B-A).sub.m-(B).sub.yO-L-F.sub.1,Q-(A).sub.x-(B-A).sub.m-(B).sub.yO-L-F.sub.1.sub.P,Q-(A).sub.x-(B-A).sub.m-(B).sub.yOH,H-(A).sub.x-(B-A).sub.m-(B).sub.yY, wherein: x is 0 or 1, Q is T when x is 0 and is chosen from T, Bn and acyl groups, for example Lev, ClAc, Fmoc, or Ac when x is 1, T is chosen from 2-naphtylmethyl (Nap), para-methoxybenzyl (PMB), 4-bromobenzyl (PBB), benzyloxymethyl acetal (BOM), 2-methoxyethoxymethylether (MEM), methoxypropyl (MOP), tetrahydropyranyl (THP), allyl (All), C.sub.1-C.sub.6 alkyl, or a silyl, T being in particular tert-butyldimethylsilyl (TBS), dimethylhexylsilyl (TDS), triisopropyl silyl (TIPS), or triethylsilyl (TES), A is ##STR00140## in particular ##STR00141## wherein: P.sub.1 is chosen from TCA, TFA, DCA, CA, Ac, benzyloxycarbamate (Cbz), Trichloroethoxycarbonyl (Troc), and Fmoc, at least one P.sub.1 and P.sub.2 being chosen from TCA, DCA, Ac, Fmoc, Troc, and, when Y is not OAll, OAlloc, P.sub.2 is H or chosen from Ac, Boc, TFA, benzyloxycarbamate (Cbz), and 2,2,2-trichloroethoxycarbonyl (Troc), P.sub.2 being H when P.sub.1 is not Ac, or P.sub.1 and P.sub.2 form together a phthalimido or a tetrachlorophthalimido (Cl.sub.4Phth) group, R.sub.2 is CO.sub.2R.sub.1 or CH.sub.2OR.sub.3, wherein R.sub.3 is Ac, benzoyl (Bz), or R.sub.3 forms with group T a benzylidene group, R.sub.1 is chosen from C.sub.1-C.sub.6 alkyl, notably Me or tert-butyl (tBu), Bn and p-methoxybenzyl (PMB) groups, R.sub.1 being in particular Bn, B is ##STR00142## in particular ##STR00143## Y is chosen from: OAll, when T is not All; Silyl ethers, in particular tert-butyldimethylsilyl ether (OTBS), dimethylhexylsilyl ether (OTDS), triethylsilyl ether (OTES), triisopropyl silyl ether (OTIPS), when T is Nap or PMB; OPMB, ONap, when OT is a silyl ether or T is All or PBB ether; p-methoxyphenyl-O (OMP or OPMP); and SR.sub.4, wherein R.sub.4 is such as the compound is a thioglycoside; L is: a single bond, a divalent C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl or C.sub.2-C.sub.12 alkynyl chain optionally interrupted by one or more heteroatoms, notably selected from an oxygen atom, a sulphur atom or a nitrogen atom, said nitrogen and sulphur atoms being optionally oxidized, and the nitrogen atom being optionally involved in an acetamide bond, or a divalent C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl or C.sub.2-C.sub.12 alkynyl chain substituted by at least one OH group, being in particular of the following formula (CH.sub.2CH.sub.2C(OH)).sub.q(CH.sub.2CH.sub.2).sub.i, wherein i is 0 or 1 and q ranges from 1 to 10, N(R.sup.a)-D-, wherein R.sup.a is H, C.sub.1-C.sub.4-alkyl, C.sub.1-C.sub.4-alkoxy, CH.sub.2C.sub.6H.sub.5, CH.sub.2CH.sub.2C.sub.6H.sub.5, OCH.sub.2C.sub.6H.sub.5, or OCH.sub.2CH.sub.2C.sub.6H.sub.5; D is C.sub.1-C.sub.7-alkylene, C.sub.1-C.sub.7-alkoxy, C.sub.1-C.sub.4-alkyl-(OCH.sub.2CH.sub.2).sub.pOC.sub.1-C.sub.4-alkyl, OC.sub.1-C.sub.4-alkyl-(OCH.sub.2CH.sub.2).sub.pOC.sub.1-C.sub.4-alkyl or C.sub.1-C.sub.7-alkoxy-R.sup.b, wherein R.sup.b is an optionally substituted aryl or an optionally substituted heteroaryl, and wherein p is 0 to 6, preferably p is 1, 2 or 3, and further preferably p is 1; Z is Z.sub.1 or F.sub.1-L.sub.2-Z.sub.2, Z.sub.1 is a terminal function or group, optionally protected, able to form a covalent bond with a carrier and/or a solid support, or a multivalent scaffold; an anchor; a mono-, oligo- or polysaccharide; or a dye or fluorescent residue. F.sub.1 is any group enabling to bond the linker L to the linker L.sub.2, F.sub.1 being in particular chosen from the C(O), C(O)C(O), C(O)C(O)NH, NHC(O)C(O), NHC(O)C(O)NH, C(O)C(H)NNH, NHC(O)C(H)NNH, ester, amide, amine, CH.sub.2, ether, thioether, imine, thio-succinimide, oxime, hydrazone, hydrazonamide, C(O)CH.sub.2NH, NHCH.sub.2C(O), triazole functions or groups, and from the following: ##STR00144## F.sub.1 being more particularly chosen from thio-succinimide, C(O)CH.sub.2NH, NHCH.sub.2C(O), and triazole functions or groups, L.sub.2 is a single bond, divalent C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl or C.sub.2-C.sub.12 alkynyl chain optionally interrupted by one or more heteroatoms, notably selected from an oxygen atom, a sulphur atom or a nitrogen atom, said nitrogen and sulphur atoms being optionally oxidized, and the nitrogen atom being optionally involved in an acetamide bond, Z.sub.2 is Z.sub.1 or F.sub.2-L.sub.3-Z.sub.1, F.sub.2 is any group enabling to bond the linker L to the linker L.sub.3, F.sub.2 being in particular chosen from the C(O), C(O)C(O), C(O)C(O)NH, NHC(O)C(O), NHC(O)C(O)NH, C(O)C(H)NNH, NHC(O)C(H)NNH, ester, amide, amine, CH.sub.2, ether, thioether, imine, thio-succinimide, oxime, hydrazone, hydrazonamide, C(O)CH.sub.2NH, NHCH.sub.2C(O), triazole functions or groups, and from the following: ##STR00145## F.sub.2 being more particularly chosen from thio-succinimide, C(O)CH.sub.2NH, NHCH.sub.2C(O), and triazole functions or groups, L.sub.3 is single bond, a divalent C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl or C.sub.2-C.sub.12 alkynyl chain optionally interrupted by one or more heteroatoms, notably selected from an oxygen atom, a sulphur atom or a nitrogen atom, said nitrogen and sulphur atoms being optionally oxidized, and the nitrogen atom being optionally involved in an acetamide bond, F.sub.1 is a precursor of F.sub.1 as defined above, F.sub.1.sub.P is a protected group F.sub.1, in particular with one or more benzyl groups, m is from 1 to n, n ranges from 1 to 50, in particular from 1 or 2 to 10, more particularly from 1 or 2 to 4 or from 3 to 8, n being notably 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, with the proviso that: when the compound is H(B).sub.x-(A-B).sub.m(A).sub.y-Y, in particular H(B).sub.x-(A-B).sub.m-(A).sub.y-OAll, and x=y=0, then m ranges from 3 to 50, in particular from 4 to 12; when the compound is Q-(B).sub.x-(A-B).sub.m-(A).sub.y-Y or Q-(A).sub.x-(B-A).sub.m(B).sub.yY, in particular Q-(B).sub.x-(A-B).sub.m-(A).sub.y-OAll or Q-(A).sub.x-(B-A).sub.m(B).sub.y-OAll, and x+y=1, then m ranges from 2 to 50, in particular from 3, 4 or 5 to 12 or 50.

    14. Compound of one of the following formulae: ##STR00146## ##STR00147## ##STR00148##

    Description

    FIGURES

    [0474] FIG. 1 presents the anti-S. sonnei LPS IgG titer induced in mice receiving three injections of glycoconjugates SonB-SonF containing 2.5 g of oligosaccharide per dose. Bleeding 3 weeks after the 3rd injection. X-axis: Glycoconjugates. Y-axis: Anti-S. sonnei LPS IgG titer. No statistically significant differences were observed between SonB, SonC and SonD. No statistically significant differences were observed between SonE and SonF. The Ab titers induced by these glycoconjugates were significantly higher than that induced by SonB, SonC and SonD. Medians are indicated (bold lines). T-test Mann Withney non parametric: *** p<0.0005.

    [0475] FIG. 2 presents the anti-S. sonnei LPS IgG titer induced in mice receiving three injections of glycoconjugates Son F-Son M containing 2 g of oligosaccharide per injection. Bleeding 1 month after the 3rd injection. X-axis: Glycoconjugates. Y-axis: Anti-S. sonnei LPS IgG titer. Medians are indicated (bold lines). T-test Mann Withney non parametric: *** p<0.0005; ** p<0.005; * p<0.05.

    [0476] FIG. 3 presents the anti-S. sonnei LPS IgG titer induced in mice receiving three doses of glycoconjugates Son H, Son K and Son M (2 g of oligosaccharide per injection). Bleeding were performed 30 days after immunization 1 (J30 imm1), 30 days after immunization 2 (J30 imm2) and 7 days and 30 days after immunization 3 (J7 imm3 and J30 imm3, respectively). X-axis: Glycoconjugates and timing of bleeding. Y-axis: Anti-S. sonnei LPS IgG titer. Medians are indicated (bold lines). T-test Mann Withney non parametric: *** p<0.0005; ** p<0.005. FIG. 4 presents the anti-S. sonnei LPS IgG titer induced in mice receiving three injections of conjugates Son W-Son Z containing 2 g or 0.5 g of oligosaccharide per injection. Bleeding was performed 3 weeks after the 3rd immunization. X-axis: Glycoconjugates and dose of oligosaccharide (2 for 2 g and 0.5 for 0.5 g, respectively). Y-axis: Anti-S. sonnei LPS TgG titer. Medians are indicated (bold lines).

    [0477] FIG. 5 presents the anti-S. sonnei LPS IgG titer induced in mice receiving three injections of conjugates Son N-Son V containing 2 g of oligosaccharide per injection. Bleeding was performed 3 weeks after the 3rd immunization. X-axis: Glycoconjugates. Y-axis: Anti-S. sonnei LPS IgG titer. Medians are indicated (bold lines). T-test Mann Withney non parametric: *** p<0.0005.

    [0478] FIG. 6 presents the anti-S. sonnei LPS IgG titer induced in mice receiving two injections of conjugates Son N, Son AA and Son BA containing 2 g of oligosaccharide per injection. Bleeding was performed 3 weeks after the 2nd immunization. X-axis: Glycoconjugates with alum (+AlH) or without. Y-axis: Anti-S. sonnei LPS IgG titer. Medians are indicated (bold lines). T-test Mann Withney non parametric: *** p<0.0005; ** p<0.005.

    [0479] FIG. 7 presents the anti-S. sonnei LPS IgG subclasses induced in mice receiving three injections of conjugate Son Y. X-axis: mouse IgG subclasses. Y-axis: Anti-S. sonnei LPS IgG subclass titer. Medians are indicated (bold lines).

    [0480] FIG. 8 presents the anti-S. sonnei LPS IgG titer induced in mice receiving two injections of conjugates Son CA-Son GA containing 2 g or 1 g of oligosaccharide per injection. Bleeding was performed 3 weeks after the 2nd immunization. X-axis: Glycoconjugates and dose of oligosaccharide (2 for 2 g and 1 for 1 g, respectively). Y-axis: Anti-S. sonnei LPS IgG titer. Medians are indicated (bold lines).

    EXAMPLES

    [0481] General Procedures [0482] Ref 1: [1] H. B. Pfister, L. A. Mulard, Org. Lett. 2014, 16, 4892-4895. [0483] Ref 2: [2] Westphal, O., and J. Jann. 1965. Bacterial lipopolysaccharides extraction with phenol-water and further application of the procedures. Meth. Carbohydr. Chem. 5: 83-91.

    [0484] Anhyd. solvents including Tol, DCM, DCE, THF, DMF, MeOH, ACN, and Py, were delivered on MS and used as received. Reactions requiring anhyd. conditions were run under an Ar atmosphere, using dried glassware. 4 MS were activated before use by heating under high vacuum. Analytical TLC was performed with silica gel 60 F254, 0.25 mm pre-coated TLC aluminium foil plates. Compounds were visualized using UV254 and/or orcinol (1 mg.Math.mL.sup.1) in 10% aq. H.sub.2SO.sub.4 with charring. Flash column chromatography was carried out using silica gel (particle size 40-63 m). RP-HPLC purification was carried out using a Kromasil 5 m C18 100 10250 mm semi-preparative column eluting with ACN in 0.08% aq. TFA, 5 mL.Math.min.sup.1, with UV (=215 nm) detection. Analytical RP-HPLC of the final compounds (=215 nm or 230 nm, ESLD) was carried out using a Kromasil 3.5 m C18 100 3150 mm analytical column, eluting with a 0-20% linear gradient of ACN in 0.08% aq. TFA over 20 min at a flow rate of 0.4 mL.Math.min.sup.1 (conditions A) or 1.0 mL.Math.min.sup.1 (conditions A), a 0-20% linear gradient of ACN in 10 mM aq. ammonium acetate over 20 min at a flow rate of 0.4 mL.Math.min.sup.1 (conditions B), or using a an Aeris Peptide 3.5 m C18 100 2.1150 mm analytical column, eluting with a 0-20% linear gradient of ACN in 0.08% aq. TFA over 20 min at a flow rate of 0.3 mL.Math.min.sup.1 (conditions C), or using a Kromasil 3.5 m C18 100 3150 mm analytical column, eluting with a 0-40% linear gradient of ACN in 0.08% aq. TFA over 20 min at a flow rate of 0.4 mL.Math.min.sup.1 (conditions D) or a 0-70% linear gradient of ACN in 0.08% aq. TFA over 20 min at a flow rate of 0.4 mL.Math.min.sup.1 (conditions E). Except for octasaccharide 4, NMR spectra were recorded at 303 K on a Bruker Avance spectrometer equipped with a BBO probe at 400 MHz (.sup.1H) and 100 MHz (.sup.13C). Spectra were recorded in CDCl.sub.3, DMSO-d6 and D.sub.2O. In the case of octasaccharide 4, NMR spectra were recorded on a 800 MHz Bruker Avance NEO equipped with a high sensitivity TCI cryogenic probe. Chemical shifts are reported in ppm () relative to the residual solvent peak in the case of CDCl.sub.3 and DMSO-d6, and to HOD and DSS (4,4-dimethyl-4-silapentane-1-sulfonic acid) in the case of D.sub.2O, at 7.28/77.0, 2.50/39.0 and 4.70/0.00 ppm for the .sup.1H and .sup.13C spectra, respectively. Coupling constants are reported in hertz (Hz). Elucidations of chemical structures were based on .sup.1H, COSY, DEPT-135, .sup.13C, HSQC, HMBC, HSQCND and NOESY spectra. Signals are reported as s (singlet), d (doublet), t (triplet), dd (doublet of doublet), q (quadruplet), dt (doublet of triplet), dq (doublet of quartet), ddd (doublet of doublet of doublet), m (multiplet). Signals can also be described as broad (prefix br), or partially overlapped (suffix po). Of the two magnetically non-equivalent geminal protons at C-6, the one resonating at lower field is denoted H-6a, and the one at higher field is denoted H-6b. Sugar residues are lettered according to the lettering of the repeating unit of the S. sonnei OAg and identified by a subscript (A, B) in the listing of signal assignments. For compounds made of multiple repeating units, residues are distinguished in the form of A/B, Ai/Bi, with A/B corresponding to the repeating unit at the reducing end. HRMS spectra were recorded on a WATERS QTOF Micromass instrument in the positive-ion electrospray ionisation (ESI.sup.+) mode. Solutions were prepared using 1:1 ACN/H.sub.2O containing 0.1% formic acid. In the case of sensitive compounds, solutions were prepared using 1:1 MeOH/H.sub.2O to which was added 10 mM ammonium acetate.

    [0485] General Procedure for Anomeric Deallylation

    [0486] [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (0.02 equiv.) was dissolved in anhyd. THF (20 mM) and stirred for 30-40 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed several times with Ar and poured into a solution of allyl glycoside (1.0 equiv.) in anhyd. THF (50-100 mM). After stirring at rt for 1-2 h, NIS (1.1 equiv.) and H.sub.2O, to reach a 1:5 H.sub.2O/THF ratio, were added. After stirring at rt for 1 h, the reaction was quenched by addition of 10% aq. sodium sulphite. The reaction mixture was concentrated and the aq. phase was extracted with DCM. The combined organic layer was washed with brine, dried over anhyd. Na.sub.2SO.sub.4 and concentrated. Purification by flash chromatography (cHex/EtOAc) yielded the expected hemiacetal as a / mixture.

    [0487] General Procedure for the Synthesis of the PTFA Glycosyl Donors

    [0488] The hemiacetal precursor (1.0 equiv.) was dissolved in acetone (0.2 M). PTFACl (1.3 equiv.) was added followed by addition of Cs.sub.2CO.sub.3 (1.1 equiv.). After stirring at rt under an Ar atmosphere until completion (estimated 2 h), the reaction mixture was filtered over a plug of Celite and washed exhaustively with anhydr. DCM. The filtrate was concentrated under reduced pressure. The crude residue was used as such in the glycosylation reaction. Purification by flash chromatography (cHex/EtOAc containing 1% Et.sub.3N) provided analytical samples.

    [0489] General procedure for hydrogenation enabling the concomitant cleavage of the benzyl esters and benzyl ethers, reduction of azides to amines and allyl aglycon to propyl aglycon, and hydrodechlorination of the trichloroacetamides into acetamides.

    [0490] Protocol 1: The oligosaccharide (50 mg) was dissolved in 2-MeTHF/isopropanol/water (1:15:3, v/v/v). 20% Pd(OH).sub.2/C (100 mg, twice the mass of oligosaccharide) was added and the reaction mixture was degassed several times and vigorously stirred under a hydrogen atmosphere for 24 h. After each 1 h, the pH of the solution was checked and the solution was neutralized by addition of 1M aq. NaHCO.sub.3 (3 equiv. per NHTCA group added within the first 6-12 h). Reaction progress was monitored by LC-MS and HRMS. In average, completion was reached within 12-24 h. The suspension was filtered by passing through a 0.2 m filter, and solids were washed repeatedly with water. Volatiles were removed under vacuum, water was added and the solution was lyophilized. The crude product was purified by preparative RP-HPLC. Fractions of interest were pooled and lyophilized to give the fully deprotected oligosaccharide as confirmed by HRMS and NMR analysis.

    [0491] Protocol 2 (H-cube, full hydrogen mode, Pressure 0 bar, column heater: 25 C., flow rate: 0.8-1.2 mL.Math.min.sup.1, 20% Pd(OH).sub.2C cartridge): 50 mg of oligosaccharide were dissolved in 2-MeTHF/isopropanol/water (1:15:3, v/v/v). The solution was subjected to hydrogenation. After each cycle, the released HCl released was quenched by addition of 1 M aq. NaHCO.sub.3 (3 equiv. per NHTCA group added within first 3-6 cycles). Reaction progress was monitored by LC-MS and HRMS analysis. In average, completion was reached after 6-12 cycles. The suspension was filtered by passing through a 0.2 m filter and solids were washed repeatedly with water. Volatiles were removed under vacuum, water was added, and the solution was lyophilized. The crude product was purified by preparative RP-HPLC. Fractions of interest were pooled and lyophilized to give the fully deprotected oligosaccharide as confirmed by HRMS and NMR analysis.

    [0492] Protocol 3 (10% PdC): 50 mg of protected oligosaccharide were dissolved in 1.0 mL 2-MeTHF and 2-MeTHF/isopropanol/water (1:10:1, v/v/v) was added to reach 0.2-0.25 mM/repeating unit. The reaction mixture was degassed several times, 10% Pd/C (twice the mass of the starting oligosaccharide) was added and the suspension was stirred vigorously under a hydrogen atmosphere (balloon) until RP-HPLC and HRMS monitoring indicated reaction completion. More 10% Pd/C was added over time if needed (up to twice the mass of the starting oligosaccharide). The pH of the solution was checked regularly and adjusted to 5-6 by addition of 1 M aq. NaHCO.sub.3 (up to 3 equiv. per NHTCA group). The suspension was filtered by passing through a pad of Celite and solids were washed repeatedly with water. Volatiles were removed under vacuum, water was added, and the solution was lyophilized. The crude product was purified by preparative RP-HPLC. Fractions of interest were pooled and lyophilized to give the fully deprotected oligosaccharide as confirmed by HRMS and NMR analysis.

    [0493] Useful Intermediates

    ##STR00085##

    [0494] Allyl 2-azido-3-O-benzyl-2-deoxy--L-altropyranoside (S2). Azide 12 (4.0 g, 9.4 mmol, 1.0 equiv.) was dissolved in AcOH (40 mL) and water (10 mL) was added. The solution was heated at 80 C. for 2 h. A TLC follow up (Tol/EtOAc 1:1) showed the absence of the starting material (R.sub.f 0.9) and the presence of a new spot (R.sub.f 0.1). After reaching rt, the reaction mixture was concentrated under vacuum. The residue was coevaporated with toluene (10 mL) twice and dried over vacuum to deliver the crude diol S2. The latter had R.sub.f 0.45 (Tol/EtOAc 1:1). .sup.1H NMR (CDCl.sub.3) 7.38-7.34 (m, 5H, H.sub.Ar), 5.99-5.90 (m, 1H, CH.sub.All), 5.36-5.31 (m, 1H, CH.sub.2All), 5.25-5.22 (m, 1H, CH.sub.2All), 4.83 (d.sub.po, 1H, J=11.2 Hz, CH.sub.2Bn), 4.80 (d.sub.po, 1H, J.sub.1,2=2.4 Hz, H-1), 4.61 (d, 1H, CH.sub.2Bn), 4.29-4.23 (m, 1H, CH.sub.2All), 4.07-4.01 (m, 1H, CH.sub.2All), 3.99 (ddd, 1H, J.sub.4,5=8.4 Hz, H-5), 3.92-3.88 (m, 1H, H-4) 3.89 (dd.sub.po, 1H, J.sub.2,3=3.5 Hz, H-2), 3.86 (dd.sub.po, 1H, J.sub.5,6b=3.6 Hz, H-6a), 3.79 (dd.sub.po, 1H, J.sub.5,6a=5.2 Hz, J.sub.6a,6b=12.3 Hz, H-6b), 3.78-3.75 (m.sub.o, 1H, H-3). .sup.13C NMR (CDCl.sub.3) 137.0 (C.sub.q, Ar), 133.6 (CH.sub.All), 129.0, 128.6, 128.2, 128.2, 128.0 (C.sub.Ar), 117.4 (CH.sub.2All), 97.9 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 76.2 (C-3), 72.5 (CH.sub.2Bn), 70.8 (C-5), 68.7 (CH.sub.2All), 64.7 (C-4), 62.9 (C-6), 59.6 (C-2). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.16H.sub.25N.sub.4O.sub.5, 353.1825; found 353.1827.

    [0495] Allyl 4,6-di-O-acetyl-2-azido-3-O-benzyl-2-deoxy--L-altropyranoside (22). The crude diol S2 (9.4 mmol theo., 1.0 equiv.) was dissolved in pyridine (40 mL) and acetic anhydride (2.2 mL, 23.6 mmol, 2.5 equiv.) was added at rt. After stirring for 3 h, a TLC analysis indicated reaction completion. Volatiles were evaporated and coevaporated with toluene (10 mL) twice, diluted with DCM (150 mL). The organic phase was washed with 1N aq. HCl (150 mL), 50% aq. NaHCO.sub.3 (150 mL) and brine (100 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. The crude was purified by flash chromatography eluting with Tol/EtOAc (4:1.fwdarw.3:1) to give diacetate 22 (3.5 g, 8.3 mmol, 88%) as a white solid. Azide 22 had R.sub.f 0.4 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.39-7.31 (m, 5H, H.sub.Ar), 5.98-5.89 (m, 1H, CH.sub.All), 5.37-5.31 (m, 1H, CH.sub.2All), 5.26-5.22 (m, 2H, H-4, CH.sub.2All), 4.72 (d, 1H, J.sub.1,2=4.4 Hz, H-1), 4.67 (dd.sub.po, 1H, J=11.6 Hz, CH.sub.2Bn), 4.29 (dd.sub.po, 1H, J.sub.5,6a=6.0 Hz, H-6a), 4.29-4.24 (m, 2H, H-5, CH.sub.2All), 4.18-4.15 (m.sub.po, 1H, H-6b), 4.09-4.04 (m, 1H, CH.sub.2All), 3.81 (dd, 1H, J.sub.2,3=8.3 Hz, H-2), 3.81 (dd, 1H, J.sub.3,4=3.8 Hz, H-3), 2.10 (s, 3H, CH.sub.3Ac), 2.08 (s, 3H, CH.sub.3Ac). .sup.13C NMR (CDCl.sub.3) 170.4, 170.0 (CO.sub.Ac), 137.1 (C.sub.q,Ar), 133.4 (CH.sub.All), 128.4, 128.0 (C.sub.Ar), 117.3 (CH.sub.2All), 98.7 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 74.6 (C-3), 72.4 (CH.sub.2Bn), 69.6 (C-5), 69.2 (CH.sub.2All), 66.3 (C-4), 62.7 (C-6), 61.5 (C-2), 20.8 (CH.sub.3Ac), 20.6 (CH.sub.3Ac). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.20H.sub.25N.sub.3O.sub.7Na, 442.1590; found 442.1597.

    [0496] Allyl 2-azido-3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy--L-altropyranoside (S3). CSA (4.1 g, 17.7 mmol, 0.5 equiv.) was added to acetal 12 (15.0 g, 35.4 mmol, 1.0 equiv.) in MeOH/DCM (4:1, 170 mL). After stirring at rt for 2 h, a TLC follow up (Tol/EtOAc 4:1) indicated reaction completion. 5% Aq. NaHCO.sub.3 (300 mL) was added followed by EtOAc (500 mL). The organic phase was separated, washed with brine (500 mL), dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The material was dried under high vacuum to give the crude diol S2 as a yellow oil. The latter was used as such in the next step.

    [0497] tert-Butyldiphenylchlorosilane (10.1 mL, 38.9 mmol, 1.1 equiv.) and imidazole (3.1 g, 46.0 mmol, 1.3 equiv.) were added to diol S2 in anhyd. DMF (180 mL) at 0 C. The reaction mixture was allowed to reach rt slowly and stirred overnight at this temperature. MeOH (10.0 mL) was added and after 30 min, volatiles were evaporated under reduced pressure. The crude material was dissolved in EtOAc (500 mL) and the organic layer was washed with 90% aq. brine (500 mL), separated, dried over Na.sub.2SO.sub.4, and concentrated to give the crude silyl ether S3. The latter had R.sub.f 0.65 (Tol/EtOAc 9:1). .sup.1H NMR (CDCl.sub.3) 7.71-7.68 (m, 4H, H.sub.Ar), 7.40-7.34 (m, 11H, H.sub.Ar), 6.00-5.90 (m, 1H, CH.sub.All), 5.35-5.29 (m, 1H, CH.sub.2All), 5.23-5.19 (m, 1H, CH.sub.2All), 4.84 (d, 1H, J.sub.1,2=4.2 Hz, H-1), 4.80 (d, 1H, J=11.4 Hz, CH.sub.2Bn), 4.62 (d, 1H, CH.sub.2Bn), 4.34-4.29 (m, 1H, CH.sub.2All), 4.09-4.04 (m, 1H, CH.sub.2All), 4.01 (ddd, 1H, H-5), 3.95 (ddd, 1H, J.sub.4,5=6.7 Hz, H-4), 3.91 (dd, 1H, J.sub.5,6b=3.3 Hz, J.sub.6a,6b=11.2 Hz, H-6a), 3.85-3.80 (m, 2H, J.sub.5,6b=5.2 Hz, H-6b, H-2), 3.77 (dd, 1H, J.sub.2,3=7.3 Hz, J.sub.3,4=3.9 Hz, H-3), 3.77 (dd, 1H, J.sub.4,OH=5.2 Hz, OH-4). .sup.13C NMR (CDCl.sub.3) 137.2, 135.7, 135.6 (C.sub.q,Ar), 133.8 (CH.sub.All), 133.2, 133.1, 129.7 (2C), 128.6, 128.1, 128.0, 127.7 (C.sub.Ar), 117.3 (CH.sub.2All), 98.2 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 76.9 (C-3), 73.4 (C-5), 72.6 (CH.sub.2Bn), 68.6 (CH.sub.2All), 65.2 (C-4), 64.5 (C-6), 60.8 (C-2), 26.8 (CH.sub.3,TBDPS), 19.2 (C.sub.TBDPS). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.32H.sub.43N.sub.4O.sub.5Si, 591.3003; found 591.2971.

    Example 1: Strategy 2.SUB.A.-NHAc,2.SUB.B.-NTCA, 4.SUB.A.-Nap

    [0498] ##STR00086##

    [0499] Disaccharide Building Block [0500] AB acceptor: from a 4.sub.A,6.sub.A-O-benzylidene A donor

    ##STR00087##

    [0501] Allyl 3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-tetrachlorophthalimido--L-altropyranoside (10). The known azide 12.sup.[1] (10.0 g, 23.6 mmol, 1.0 equiv.) was dissolved in anhyd. THF (95 mL). Zn dust (12.3 g, 189 mmol, 8.0 equiv.) and AcOH (10.8 mL, 189 mmol, 8.0 equiv.) were added at rt. The reaction mixture was stirred vigorously for 1 h. A TLC follow up (Tol/EtOAc 1:1) showed the formation of a highly polar product and the full consumption of the starting 12 (R.sub.f 0.9). The suspension was filtered through a pad of Celite and washed with DCM (100 mL) twice. The filtrate was washed with satd. aq. NaHCO.sub.3. The organic layer was dried over anhyd. Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue, corresponding to the known amine 13.sup.[1] was dried under high vacuum for 2 h. The crude material (9.6 g) was subjected to the next step.

    [0502] Tetrachlorophthalic anhydride (4.05 g, 14.1 mmol, 0.6 equiv.) was added to the crude 13 (9.38 g, 23.6 mmol theo.) stirred in anhyd. DCM (100 mL) at rt under an Ar atmosphere. After 30 min, Et.sub.3N (3.2 mL, 23.6 mmol, 1.0 equiv.) followed by more TCPO (4.05 g, 14.1 mmol, 0.6 equiv.) were added. The reaction mixture was stirred for another 30 min at rt, at which time a TLC follow up (EtOAc) revealed the presence of a polar product (R.sub.f 0.0) and absence of 13 (R.sub.f 0.15). Volatiles were eliminated under reduced pressure and the residue was dried under high vacuum for 1 h. The crude was dissolved in anhyd. Py (90 mL) and Ac.sub.2O (11.1 mL, 118 mmol, 5.0 equiv.) was added at rt. The mixture was heated to 80 C. for 10 min, at which time a TLC follow up indicated completion. Volatiles were eliminated under reduced pressure and coevaporated with toluene (40 mL) twice. The crude was diluted with DCM (200 mL) and washed with 1N aq. HCl (300 mL), satd. aq. NaHCO.sub.3 (300 mL) and brine (250 mL). The DCM layer was dried over Na.sub.2SO.sub.4, filtered and concentrated. The crude was purified by flash chromatography (cHex/EtOAc, 98:2 to 90:10) to give the fully protected 10 (13.0 g, 18.8 mmol, 83%) as a dense yellowish oil. Allyl glycoside 10 had R.sub.f 0.65 (Tol/EtOAc 10:1). .sup.1H NMR (CDCl.sub.3) 7.53-7.05 (m, 10H, H.sub.Ar), 5.88-5.80 (m, 1H, CH.sub.All), 5.65 (s, 1H, H.sub.Bzl), 5.28-5.23 (m, 1H, CH.sub.2All), 5.16-5.12 (m.sub.po, 1H, CH.sub.2All), 5.14 (d.sub.po, 1H, J.sub.1,2=4.0 Hz, H-1), 4.85 (d, 1H, J=12.5 Hz, CH.sub.2Bn), 3.92 (t, 1H, J.sub.2,3=4.0 Hz, H-2), 4.71 (d, 1H, CH.sub.2Bn), 4.52-4.42 (m, 2H, H-5, H-6a), 4.34 (dd.sub.po, 1H, J.sub.3,4=4.4 Hz, J.sub.4,5=9.6 Hz, H-4), 4.27-4.22 (m, 1H, CH.sub.2All), 4.10 (t, 1H, H-3), 4.03-3.97 (m, 1H, CH.sub.2All), 3.88 (t, 1H, J.sub.5,6b=J.sub.6a,6b=10.0 Hz, H-6b). .sup.13C NMR (CDCl.sub.3), 162.3 (CO.sub.NTCP), 140.3, 138.1, 137.6, 137.6 (C.sub.q, Ar), 133.6 (CH.sub.All), 129.8, 129.0, 128.2, 128.0 (2C), 127.2, 126.9, 126.4 (C.sub.Ar), 117.3 (CH.sub.2All), 101.8 (C.sub.Bzl), 95.4 (C-1.sub.A, .sup.1J.sub.C,H=171 Hz), 76.4 (C-4), 73.1 (C-3), 72.8 (CH.sub.2Bn), 69.7 (C-6), 68.4 (CH.sub.2All), 59.8 (C-5), 55.5 (C-2). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.31H.sub.25Cl.sub.4NO.sub.7Na, 686.0283; found 686.0284.

    [0503] Allyl 3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-N-(9-fluorenylmethoxycarbonyl)--L-altropyranoside (11). A solution of azide 12 (2.4 g, 5.6 mmol, 1.0 equiv.) in THF (30 mL) was added triphenylphosphine (1.63 g, 6.2 mmol, 1.1 equiv.) and H.sub.2O (3.0 mL, 169 mmol, 30 equiv.). The reaction mixture was stirred overnight at 60 C., cooled to rt, concentrated under reduced pressure and coevaporated with toluene (10 mL) twice. The crude amine 13 in anhyd. DCM (28 mL) was stirred over freshly activated MS 4 for 30 min at rt under an Ar atmosphere. After cooling to 0 C., NaHCO.sub.3 (953 mg, 11.3 mmol, 2.0 equiv.), DMAP (69 mg, 567 mol, 0.1 equiv.) and FmocCl (1.7 g, 6.8 mmol, 1.2 equiv.) were added. After stirring for 1 h at this temperature, a TLC analysis (cHex/EtOAc 9:1) revealed the conversion of intermediate 13 (R.sub.f 0.45) into a less polar product (R.sub.f 0.7). The reaction mixture was filtered and solids were washed with DCM (20 mL) twice. The filtrate was concentrated in vacuo and the residue was purified by flash chromatography using cHex/EtOAc (20:1-15:1) to give the Fmoc derivative 11 (3.1 g, 5.0 mmol, 88%) as a white solid. The fully protected 11 had .sup.1H NMR (CDCl.sub.3) 7.81-7.78 (m, 2H, H.sub.Ar), 7.65-7.60 (m, 2H, H.sub.Ar), 7.51-7.26 (m, 15H, H.sub.Ar), 5.98-5.89 (m, 1H, CH.sub.All), 5.57 (s, 1H, H.sub.Bzl), 5.37-5.31 (m, 1H, CH.sub.2All), 5.22-5.20 (m, 1H, CH.sub.2All), 4.96 (d, J.sub.2,NH=8.8 Hz, NH), 4.88 (d, 1H, J=12.6 Hz, CH.sub.2Bn), 4.83 (d, 1H, CH.sub.2Bn), 4.73 (s, 1H, H-1), 4.59-4.48 (m, 3H, H-5, CH.sub.2Fmoc), 4.32 (dd, 1H, J.sub.5,6a=5.6 Hz, J.sub.6a,6b=10.4 Hz, H-6a), 4.29-4.21 (m, 3H, CH.sub.Fmoc, H-2, CH.sub.2All), 4.04-3.99 (m, 1H, CH.sub.2All), 3.90 (brs, 1H, H-3), 3.73 (t.sub.po, 1H, J.sub.5,6b=10.4 Hz, H-6b), 3.70 (brd.sub.po, 1H, J.sub.4,5=8.8 Hz, H-4). .sup.13C NMR (CDCl.sub.3) 155.2 (CO.sub.NHFmoc), 144.3 143.6, 141.5, 141.4, 138.7, 137.6 (C.sub.q,Ar), 133.6 (CH.sub.All), 129.0, 128.2, 128.0, 127.8, 127.5, 127.3, 127.1, 127.0, 126.2, 124.7, 120.0 (C.sub.Ar), 117.2 (CH.sub.2All), 102.3 (C.sub.Bzl), 98.9 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 77.0 (C-4), 74.2 (C-3), 72.1 (CH.sub.2Bn), 69.2 (C-6), 68.4, (CH.sub.2All), 66.6 (CH.sub.2Fmoc), 58.7 (C-5), 52.1 (C-2), 47.3 (CH.sub.Fmoc). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.38H.sub.37NO.sub.7Na, 642.2468; found 642.2464.

    [0504] 3-O-Benzyl-4,6-O-benzylidene-2-deoxy-2-tetrachlorophthalimido-/-L-altropyranose (14). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (38 mg, 45 mol, 0.02 equiv.) was dissolved in anhyd. THF (5.0 mL) and the red solution was stirred for 30 min under an H.sub.2 atmosphere. The resulting light yellow solution was degassed several times with Ar and poured into a solution of allyl glycoside 10 (1.5 g, 2.26 mmol, 1.0 equiv.) in anhyd. THF (25 mL). After stirring at rt for 2 h, a TLC follow up (cHex/EtOAc; 15:1) showed the absence of the starting 10 (R.sub.f 0.3) and the presence of a less polar spot (R.sub.f 0.35). Iodine (1.14 g, 6.1 mmol, 1.05 equiv.) immediately followed by NaHCO.sub.3 (571 mg, 6.78 mmol, 3.0 eqv.) in water (5 mL) were added. After stirring at rt for 1 h, the reaction was quenched by addition of 10% aq. Na.sub.2SO.sub.3. Volatiles were evaporated and the aq. layer was extracted with DCM (100 mL) twice. Purification by flash chromatography using cHex/EtOAc (10:1.fwdarw.8:1) gave the expected hemiacetal 14 (610 mg, 0.98 mmol, 43%) as a white floppy solid corresponding mainly to a single anomer. The later had R.sub.f 0.15 (Tol/EtOAc; 10:1). .sup.1H NMR (CDCl.sub.3) 7.55-7.52 (m, 2H, H.sub.Ar), 7.44-7.38 (m, 3H, H.sub.Ar), 7.11-7.09 (m, 2H, H.sub.Ar), 6.94 (t, 2H, J=7.6 Hz, H.sub.Ar), 6.78-6.74 (m, 1H, H.sub.Ar), 6.35 (dd, 1H, J.sub.1,OH=8.8 Hz, J.sub.1,2=4.8 Hz, H-1), 5.61 (s, 1H, H.sub.Bzl), 4.84 (d, 1H, J=12.4 Hz, CH.sub.2Bn), 4.46 (dd, 1H, J.sub.5,6a=5.2 Hz, J.sub.6a,6b=10.4 Hz, H-6a), 4.39 (d, 1H, CH.sub.2Bn), 4.30-4.24 (m, 2H, H-3, H-5), 4.02 (dd, 1H, J.sub.2,3=3.2 Hz, H-2), 3.87-3.81 (m, 3H, H-6b, H-4, OH). .sup.13C NMR (CDCl.sub.3), 162.9 (CO.sub.NTCP), 139.6, 137.7, 137.3 (C.sub.q,Ar), 129.4, 129.1, 128.9, 128.3, 127.9, 127.0 (2C), 126.1 (C.sub.Ar), 102.0 (C.sub.Bzl), 91.1 (C-1.sub.A, .sup.1J.sub.C,H=176 Hz), 79.3 (C-4), 74.0 (CH.sub.2Bn), 73.9 (C-3), 69.0 (C-6), 64.4 (C-5), 58.4 (C-2). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.28H.sub.21Cl.sub.4NO.sub.7Na, 645.9970; found 645.9964.

    [0505] 3-O-Benzyl-4,6-O-benzylidene-2-deoxy-2-N-(9-fluorenylmethoxycarbonyl)-/-L-altropyranose (15). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (71 mg, 84 mol, 0.02 equiv.) in anhyd. THF (5.0 mL) was stirred under an H.sub.2 atmosphere for 1 h at rt. The resulting yellow solution was degassed several times with Ar and transferred by use of a cannula into a solution of allyl glycoside 11 (2.6 g, 4.1 mmol, 1.0 equiv.) in anhyd. THF (40 mL). After stirring at rt for 1 h, NIS (1.03 g, 4.6 mmol, 1.1 equiv.) and H.sub.2O (8 mL) were added and the reaction mixture was stirred for an additional hour. At completion, the reaction was quenched by addition of 10% aq. Na.sub.2SO.sub.3, and volatiles were eliminated. The aq. phase was extracted with DCM (40 mL) twice. The combined organic phases were dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. Purification of the residue by flash chromatography using Tol/EtOAc (5:1.fwdarw.4:1) yielded the expected 15 (2.2 g, 3.7 mmol, 90%) as a white solid. Hemiacetal 15 was isolated as a 7:3 / mix and had R.sub.f 0.2 (Tol/EtOAc, 4:1). .sup.1H NMR (Partial assignment, CDCl.sub.3) 7.79-7.77 (m, 2.7H, H.sub.Ar), 7.76-7.62 (m, 2.6H, NH, H.sub.Ar), 7.54-7.51 (m, 2.7H, H.sub.Ar), 7.43-7.28 (m, 19.5H, H.sub.Ar), 7.22-7.18 (m, 5.0H, H.sub.Ar), 5.60 (s, 0.4H, H.sub.Bzl,), 5.57 (s, 1H, H.sub.Bzl,), 5.35-5.30 (m, 1H, H-1.sub.), 5.01-4.90 (m, 1.9H, H-1, CH.sub.2Bn, CH.sub.2Bn), 4.78-4.75 (m, 1.4H, CH.sub.2Bn, CH.sub.2Bn), 4.58-4.55 (m, 2.6H, CH.sub.2Fmoc,), 4.34-4.30 (m, 1H, H-6a.sub.), 4.26-4.13 (m, 5H, H-2.sub., H-2.sub., H-3.sub., H-3.sub., H-5.sub., H-5.sub., CH.sub.Fmoc,, CH.sub.Fmoc,), 3.83-3.69 (m, 2.9H, H-4.sub., H-4.sub., H-6b.sub.), 3.62 (brs, 0.9H, OH.sub.). .sup.13C NMR (CDCl.sub.3), 156.4 (CO.sub.Fmoc), 143.6, 143.5, 141.4, 141.3, 138.3, 137.8, 137.4, 137.0 (C.sub.q,Ar), 129.2, 129.1, 129.0, 128.5, 128.3 (2C), 128.2, 128.0, 127.8, 127.6, 127.1, 126.1 (2C), 125.3, 124.9, 124.8, 124.7, 120.0 (C.sub.Ar), 102.6 (C.sub.Bzl), 102.3 (C.sub.Bzl), 94.7 (C-1.sub., .sup.1J.sub.C,H=175.6 Hz), 92.0 (C-1.sub.A, .sup.1J.sub.C,H=170.6 Hz), 77.3 (C-4.sub., C-4.sub.), 75.3 (C-3.sub.), 75.3 (C-3.sub.), 74.5 (CH.sub.2Bn, ), 73.5 (CH.sub.2Bn, ), 69.3 (C-6.sub.), 68.9 (C-6.sub.), 66.8 (CH.sub.2Fmoc,), 66.8 (CH.sub.2Fmoc,), 64.5 (C-5.sub., C-5.sub.), 54.6 (C-2.sub.), 52.7 (C-2.sub.), 47.3 (CH.sub.Fmoc,, CH.sub.Fmoc,). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.35H.sub.33NO.sub.7Na, 602.2155; found 602.2153.

    [0506] 3-O-Benzyl-4,6-O-benzylidene-2-deoxy-2-tetrachlorophthalimido-/-L-altropyranosyl (N-phenyl)trifluoroacetimidate (16). Hemiacetal 14 (2.85 g, 4.2 mmol, 1.0 equiv.) was dissolved in acetone (40 mL) and PTFACl (855 L, 5.5 mmol, 1.3 equiv.) was added followed by addition of Cs.sub.2CO.sub.3 (1.68 g, 5.1 mmol, 1.1 equiv.). After stirring for 2 h at rt, a TLC follow (Tol/EtOAc 10:1) showed the presence of a less polar spot (R.sub.f 0.7) and only traces of hemiacetal 14. The reaction mixture was filtered over a pad of Celite and washed with acetone (10 mL) twice. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography (cHex/EtOAc 95:5.fwdarw.93:7 containing 1% Et.sub.3N) to give donor 16 as a white solid (2.75 g, 3.46 mmol, 80%). The latter, obtained as a 10:3 mixture of two anomers, had R.sub.f 0.3 (cHex/EtOAc 10:1). .sup.1H NMR (partial assignment, CDCl.sub.3) 7.76 (brs, 0.8H, H.sub.Ar), 7.59 (d, 0.8H, J=7.6 Hz, HM), 7.52-7.08 (m, 18.4H, H.sub.Ar), 6.98 (d, 0.4H, J=7.2 Hz, HM), 6.82 (d, 1.6H, J=7.2 Hz, H.sub.Ar), 5.50 (s, 0.8H, CH.sub.Bzl), 5.24 (s, 0.2H, CH.sub.Bzl), 5.01-4.96 (m, 1.8H), 4.76-4.66 (m, 1.3H), 4.35 (dd, 0.8H, J=5.2 Hz, J=10.8 Hz), 4.25 (dd, 0.4H), 3.94 (t, 0.8H, J=8.0 Hz, J=8.4 Hz), 3.87-3.81 (m, 0.8H), 3.79-3.71 (m, 0.4H), 3.64-3.51 (m, 0.8H), 3.46-3.43 (m, 0.8H). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.36H.sub.25C.sub.14F.sub.3N.sub.2O.sub.7Na, 817.0266; found 817.0242.

    [0507] Allyl 3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-tetrachlorophthalimido--L-altropyranosyl-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (17), Allyl 3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-tetrachlorophthalimido--L-altropyranosyl-(1-3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (18) and 3-O-Benzyl-4,6-O-benzylidene-2-deoxy-2-tetrachlorophthalimido-L-altral (S1). A mixture of acceptor 8 (600 mg, 1.61 mmol, 1.0 equiv.) and PTFA donor 16 (1.4 g, 1.77 mmol, 1.1 equiv.) in anhyd. DCE (20 mL) was stirred with freshly activated 4 MS (1.0 g) for 30 min at rt under an Ar atmosphere. The reaction mixture was cooled to 20 C. and TMSOTf (19 L, 10 mol, 0.06 equiv.) was added slowly. After stirring for 30 min at 20 C., at which time a TLC follow up indicated donor consumption, Et.sub.3N was added, the suspension was filtered and the filtrate was concentrated in vacuo. Flash chromatography using cHex/EtOAc (15:1.fwdarw.10:1) gave the unwanted -isomer 17 (960 mg, 0.98 mmol, 61%) as a white solid, along with the desired -isomer 18 (150 mg, 0.15 mol, 10%), and the elimination product S1 (220 mg, 0.36 mmol, 20% wrt 16). The -isomer 17 had R.sub.f 0.35 (cHex/EtOAc 10:1). .sup.1H NMR (CDCl.sub.3) 7.53-7.39 (m, 5H, H.sub.Ar), 7.09 (dd, 2H, J=1.0, 8.0 Hz, H.sub.Ar), 6.95 (t, 2H, J=7.6 Hz, H.sub.Ar), 6.79-6.73 (m, 2H, H.sub.Ar, NH), 6.20 (d, 1H, J.sub.1,2=8.5 Hz, H-1.sub.A), 5.90-5.80 (m, 1H, CH.sub.All), 5.58 (s, 1H, H.sub.Bzl), 5.28-5.23 (m, 1H, CH.sub.2All), 5.18-5.14 (m, 1H, CH.sub.2All), 4.82 (d, 1H, J=12.5 Hz, CH.sub.2Bn), 4.77 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.45 (dd, 1H, J.sub.5,6a=5.1 Hz, J.sub.6a,6b=10.2 Hz, H-6a.sub.A), 4.41 (dd.sub.po, 1H, J.sub.3,4=3.4 Hz, J.sub.2,3=10.9 Hz, H-3.sub.B), 4.40 (d.sub.po, 1H, CH.sub.2Bn), 4.35-4.30 (m, 1H, CH.sub.2All), 4.28 (brt, H-3.sub.A), 4.24 (ddd, 1H, J.sub.4,5=9.6 Hz, J.sub.5,6=4.8 Hz, H-5.sub.A), 4.22 (dd, 1H, J.sub.2,3=3.1 Hz, H-2.sub.A), 4.07-4.02 (m, 1H, CH.sub.2All), 3.97 (brd, 1H, H-4.sub.B), 3.86 (t, 1H, J.sub.5,6b=10.4 Hz, H-6.sub.bA), 3.80 (dd, 1H, J.sub.4,5=9.6 Hz, J.sub.3,4=2.3 Hz, H-4.sub.A), 3.72-3.64 (m, 2H, H-2.sub.B, H-5.sub.B), 1.36 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 163.8 (CO.sub.NHTCA), 162.7, 161.6 (2C, CO.sub.NTCP), 139.9, 139.5, 137.3, 137.1 (C.sub.q, Ar), 133.5 (CH.sub.All), 129.6, 129.3, 129.2, 128.9, 128.3, 128.0, 127.7, 127.1, 126.2, 126.1 (C.sub.Ar), 117.7 (CH.sub.2All), 102.0 (C.sub.Bzl), 98.8 (C-1.sub.B, .sup.1J.sub.C,H=161 Hz), 96.0 (C-1.sub.A, .sup.1J.sub.C,H=173 Hz), 92.8 (CCl.sub.3), 79.2 (C-4.sub.A), 76.9 (C-3.sub.B), 74.6 (CH.sub.2Bn), 73.9 (C-3.sub.A), 69.9 (CH.sub.2All), 69.3 (C-5.sub.B), 69.2 (C-6.sub.A), 64.5 (C-5.sub.A), 63.5 (C-4.sub.B), 57.1 (C-2.sub.A), 53.7 (C-2.sub.B), 17.4 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.39H.sub.34C.sub.17N.sub.5O.sub.10Na, 1000.0023; found 1000.0015.

    [0508] The -isomer 18 had R.sub.f 0.3 (cHex/EtOAc 10:1). .sup.1H NMR (CDCl.sub.3) 7.54-7.51 (m, 2H, H.sub.Ar), 7.48-7.37 (m, 3H, HA), 7.28 (d, 2H, J=6.8 Hz, H.sub.Ar), 7.12 (t.sub.po, 3H, J=8.0 Hz, H.sub.Ar), 6.97 (t.sub.po, 1H, J=7.2 Hz, H.sub.Ar), 6.79 (d, 1H, J.sub.2,NH=6.4 Hz, NH.sub.B), 5.86-5.76 (m, 1H, CH.sub.All), 5.63 (s, 1H, H.sub.Bzl), 5.35 (d, 1H, J.sub.1,2=5.6 Hz, H-1.sub.A), 5.24-5.18 (m, 1H, CH.sub.2All), 5.16-5.12 (m, 1H, CH.sub.2All), 4.82 (d, 1H, J=12.0 Hz, CH.sub.2Bn), 4.81 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B), 4.74 (dd, 1H, J.sub.2,3=4.8, Hz, H-2.sub.A), 4.64 (dd, 1H, J.sub.3,4=3.6 Hz, J.sub.2,3=10.8 Hz, H-3.sub.B), 4.58-4.46 (m, 2H, H-5.sub.A, CH.sub.2Bn), 4.50 (dd, 1H, J.sub.5,6a=5.2 Hz, J.sub.6a,6b=10.4 Hz, H-6a.sub.A), 4.32-4.27 (m, 1H, CH.sub.2All), 4.21 (dd, 1H, J.sub.3,4=4.8 Hz, J.sub.4,5=8.8 Hz, H-4.sub.A), 4.12 (brt, 1H, H-3.sub.A), 4.02-3.97 (m, 1H, CH.sub.2All), 3.89 (brd, 1H, J.sub.3,4=2.8 Hz, H-4.sub.B), 3.85 (t, 1H, J.sub.5,6b=10.1 Hz, H-6b.sub.A), 3.71 (dq.sub.po, 1H, H-5.sub.B), 3.53-3.46 (m, 1H, H-2.sub.B), 1.39 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 162.5 (CO.sub.NHTCA), 161.7 (CO.sub.NTCP), 140.3, 137.9, 137.3 (C.sub.q,Ar), 133.4 (CH.sub.All), 129.9, 129.1, 128.4, 128.2, 127.9, 127.1, 126.9, 126.2 (C.sub.Ar), 118.0 (CH.sub.2All), 101.7 (C.sub.Bzl), 97.7 (C-1.sub.A, .sup.1J.sub.C,H=173 Hz), 97.3 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.1 (CCl.sub.3), 76.3 (C-4.sub.A), 76.1 (C-3.sub.B), 73.0 (CH.sub.2Bn), 72.0 (C-3.sub.A), 70.2 (CH.sub.2All), 69.6 (C-6.sub.A), 69.1 (C-5.sub.B), 65.8 (C-4.sub.B), 60.8 (C-5.sub.A), 55.7 (C-2.sub.A), 55.7 (C-2.sub.B), 17.5 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.39H.sub.38Cl.sub.7N.sub.6O.sub.10, 995.0469; found 995.0437.

    [0509] Altral S1 had R.sub.f 0.65 (cHex/EtOAc 10:1). .sup.1H NMR (CDCl.sub.3) 7.58-7.56 (m, 2H, H.sub.Ar), 7.45-7.40 (m, 3H, H.sub.Ar), 7.15 (d.sub.po, 2H, J=7.2 Hz, H.sub.Ar), 6.96-6.92 (m.sub.po, 2H, H.sub.Ar), 6.79-6.75 (m, 1H, H.sub.Ar), 6.61 (s, 1H, H-1), 5.67 (s, 1H, H.sub.Bzl), 4.83 (d, 1H, J=12.6 Hz, CH.sub.2Bn), 4.60 (dd, 1H, J.sub.5,6a=5.3 Hz, J.sub.6a,6b=10.4 Hz, H-6a), 4.50 (dt, 1H, J.sub.5,6b=10.3 Hz, J.sub.4,5=5.3 Hz, H-5), 4.39 (d, 1H, J.sub.3,4=5.3 Hz, H-3), 4.38 (d, 1H, CH.sub.2Bn), 4.21 (dd, 1H, H-4), 3.96 (t, 1H, H-6b). .sup.13C NMR (CDCl.sub.3) 162.5 (CO.sub.NTCP), 147.3 (C-1), 139.9, 138.6, 137.2 (C.sub.q,Ar), 129.7, 129.2, 128.7, 128.3, 127.9, 127.1, 126.9, 126.2 (C.sub.Ar), 108.4 (C-2), 101.7 (C.sub.Bzl), 77.9 (C-4), 74.0 (CH.sub.2Bn), 68.3 (C-6), 67.6 (C-3), 65.1 (C-5). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.28H.sub.23C.sub.14N.sub.2O.sub.6, 623.0310; found 623.0295.

    [0510] 3-O-Benzyl-4,6-O-benzylidene-2-deoxy-2-N-(9-fluorenylmethoxycarbonyl)-/-L-altropyranosyl (N-phenyl)trifluoroacetimidate (19). Hemiacetal 15 (170 mg, 293 mol, 1.1 equiv.) was dissolved in acetone (6 mL). PTFACl (60 L, 382 mol, 1.3 equiv.) was added followed by addition of Cs.sub.2CO.sub.3 (105 mg, 323 mol, 1.1 equiv.). After stirring at rt for 1 h, a TLC analysis indicated reaction completion. The reaction mixture was filtered over a pad of Celite, and solids were washed with DCM (5 mL) twice. Volatiles were evaporated under reduced pressure and the residue was purified by flash chromatography eluting with cHex/EtOAc (98:2.fwdarw.90:10) to give the desired 19 (200 mg, 273 mol, 90%) as an off-white solid. The constrained PTFA donor had R.sub.f 0.8 (Tol/EtOAc 9:1). .sup.1H NMR (cc anomer, CDCl.sub.3) 7.68-7.50 (m, 18H, H.sub.Ar), 5.67 (d, 1H, J=3.6 Hz, H-1), 5.50 (s, 1H, H.sub.Bzl), 4.67 (brs, 1H, CH.sub.2Fmoc), 4.54 (brs, 1H, CH.sub.2Bn), 4.50 (dd.sub.po, 1H, CH.sub.2Fmoc), 437-4.33 (m, 1H, H-6a), 4.27-4.10 (m, 3H, H-5, H-3, CH.sub.Fmoc), 3.70-3.58 (m, 2H, H-6b, H-2) 3.60 (dd.sub.po, 1H, J.sub.3,4<1.0 Hz, J.sub.4,5=8.8 Hz, H-4). .sup.13C NMR (CDCl.sub.3) 153.4 (CO.sub.NFmoc), 141.3, 139.7, 137.8, 137.4 (C.sub.q,Ar), 129.4, 129.1, 128.5, 128.3, 128.2 (2C), 127.8, 127.7, 127.6, 127.5, 127.3, 127.2, 127.0, 126.3, 126.1, 124.6, 124.3, 122.5, 120.4, 120.3, 120.2 (C.sub.Ar), 102.6 (C.sub.Bzl), 95.8 (C-1.sub.A, .sup.1J.sub.C,H=182 Hz), 75.7 (C-4.sub.), 73.7 (CH.sub.2Bn), 71.6 (C-3), 69.0 (C-6), 67.2 (CH.sub.2Fmoc), 63.7 (C-5.sub.), 61.2 (C-2.sub.), 47.0 (CH.sub.Fmoc). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.43H.sub.38F.sub.3N.sub.2O.sub.7, 751.2631; found 751.2665.

    [0511] Allyl 3-O-benzyl-4,6-O-benzylidene-2-deoxy-2-N-(9-fluorenylmethoxycarbonyl)--L-altropyranosyl-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (21). Hemiacetal 15 (343 mg, 59 mol, 1.1 equiv.) was dissolved in acetone (10 mL) and cooled to 0 C. Trichloroacetonitrile (855 L, 2.36 mmol, 4.0 equiv.) was added followed by addition of K.sub.2CO.sub.3 (163 mg, 1.18 mmol, 2.0 equiv.). After stirring for 4 h at 0 C., the reaction mixture was filtered over a pad of Celite, washed with DCM (5 mL) twice. The filtrate was concentrated under reduced pressure to give the crude trichloroacetimidate 20.

    [0512] The crude 20 (1.1 equiv. theo) was mixed with acceptor 8 (200 mg, 538 mol, 1.0 equiv.), coeveporated with toluene repeatedly, and dried under high vacuum for 2 h. The mixture was dissolved in anhyd. DCM (10 mL) and stirred with freshly activated MS 4 (500 mg) for 45 min under an Ar atmosphere before the temperature was set to 15 C. TMSOTf (7 L, 30 mol, 0.05 equiv.) was added slowly. After stirring at this temperature for 30 min, a TLC analysis (Tol/EtOAc 3:1) showed the presence of a new spot (R.sub.f 0.4) close to those featuring acceptor 8 (R.sub.f 0.35) and hemiacetal 15 (R.sub.f 0.32). Et.sub.3N (10 L) was added and the suspension was filtered over a fitted funnel. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with Tol/EtOAc (5:1.fwdarw.4:1) to give the desired disaccharide 21 (200 mg, 214 mol, 62%). The coupling product had .sup.1H NMR (CDCl.sub.3) 7.80-7.77 (m, 2H, H.sub.Ar), 7.59-7.57 (d.sub.po, 2H, H.sub.Ar), 7.52-7.29 (m, 15H, H.sub.Ar), 6.90 (d, J.sub.2,NH=6.2 Hz, NH.sub.B), 5.90-5.80 (m, 1H, CH.sub.All), 5.56 (s, 1H, CH.sub.Bzl), 5.29-5.24 (m, 1H, CH.sub.2All), 5.20-5.17 (m, 1H, CH.sub.2All), 4.99 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B), 4.87 (d.sub.po, 1H, J=11.8 Hz, CH.sub.2Bn), 4.84 (d.sub.po, 1H, J=7.2 Hz, NH.sub.A), 4.80 (d.sub.po, 1H, CH.sub.2Bn), 4.77 (s.sub.po, 1H, H-1.sub.A), 4.64-4.58 (m, 2H, H-3.sub.B, H-5.sub.A), 4.52 (brd, 2H, J=5.8 Hz, CH.sub.2, NHFmoc), 4.36-4.31 (m, 2H, H-6a.sub.A, CH.sub.2All), 4.24 (brd, 1H, J.sub.2,NH=7.2 Hz, H-2.sub.A), 4.20 (t, 1H, J=6.0 Hz, CH.sub.NHFmoc), 4.09-4.04 (m, 1H, CH.sub.2All), 3.87 (brs, 1H, H-3.sub.A), 3.78 (brq, 1H, H-5.sub.B), 3.72 (t, 1H, J.sub.5,6b=J.sub.6a,6b=10.5 Hz, H-6b.sub.A), 3.69 (brd.sub.po, 1H, J.sub.3,4=3.0 Hz, H-4.sub.B), 3.67 (brd.sub.po, 1H, J.sub.4,5=8.0 Hz, H-4.sub.A), 3.49-3.43 (m, 1H, H-2.sub.B), 1.42 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 162.2 (CO.sub.NTCA), 154.9 (CO.sub.NFmoc), 143.5, 141.4, 141.3, 138.9, 137.3 (C.sub.q,Ar), 133.5 (CH.sub.All), 129.1, 128.2, 128.0, 127.8, 127.4, 127.2, 127.1, 126.2, 124.8, 124.7, 120.0 (C.sub.Ar), 117.9 (CH.sub.2All), 102.3 (C.sub.Bzl), 101.6 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 97.2 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.0 (CCl.sub.3), 76.7 (C-4.sub.A), 75.6 (C-3.sub.B), 74.0 (C-3.sub.A), 72.3 (CH.sub.2Bn), 70.2 (CH.sub.2All), 69.7 (C-5.sub.B), 69.1 (C-6.sub.A), 66.6 (CH.sub.2Fmoc), 65.9 (C-4.sub.B), 59.7 (C-5.sub.A), 56.1 (C-2.sub.B), 52.4 (C-2.sub.A), 47.3 (CH.sub.NHFmoc), 17.4 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.46H.sub.50Cl.sub.3N.sub.6O.sub.7, 951.2654; found 951.2694.

    [0513] Use of a A Donor Whereby Protecting Groups as the 4.sub.A-OH and 6.sub.A-OH are Identical

    ##STR00088##

    [0514] Allyl 4,6-di-O-acetyl-3-O-benzyl-2-deoxy-2-tetrachlorophthalimido--L-altropyranoside (23). Zn dust (5.6 g, 85.8 mmol, 8.0 equiv.) and AcOH (4.9 mL, 85.8 mmol, 8.0 equiv.) were added to a solution of azide 22 (4.5 g, 10.7 mmol, 1.0 equiv.) in THF (70 mL) at rt. The reaction mixture was stirred for 2 h at which point a TLC follow up (Tol/EtOAc 1:1) revealed the absence of any remaining 22 (R.sub.f 0.8) and the presence of a more polar product. The mixture was filtered over a pad of Celite and solids were washed with DCM (100 mL) twice. The combined organic phases were washed with satd. aq. NaHCO.sub.3 (200 mL) and brine (200 mL). The organic phase was dried over Na.sub.2SO.sub.4, filtered, and concentrated. The residue was dried under high vacuum for 3 h and subjected as such to the next step. The crude amine intermediate had HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.20H.sub.28NO.sub.7, 394.1866; found 394.1856.

    [0515] Tetrachlorophthalic anhydride (3.68 g, 12.8 mmol, 1.2 equiv.) was added to a solution of the crude intermediate in DCM (40 mL) and the solution was stirred for 30 min at rt. Et.sub.3N (1.79 mL, 12.8 mmol, 1.2 equiv.) was added and the reaction mixture was stirred for another 30 min. Volatiles were eliminated under reduced pressure and the residue was dried under high vacuum for 1 h. The crude material was dissolved in pyridine (50 mL) and Ac.sub.2O (5.0 mL, 53.6 mmol, 5.0 equiv.) was added at 0 C. The mixture was heated to 80 C. for 10 min. A TLC follow up (Tol/EtOAc 9:1) showed the formation of a product (R.sub.f 0.7) slightly more polar than azide 22 (0.75). After it reached rt, the reaction mixture was concentrated and coevaporated with Toluene (15 mL) twice. The residue was diluted with DCM (100 mL) and washed with 1N aq. HCl (200 mL), satd. aq. NaHCO.sub.3 (200 mL) and brine (200 mL). The DCM layer was dried over Na.sub.2SO.sub.4, filtered, concentrated and the residue was purified by flash chromatography eluting with cHex/EtOAc (12:1.fwdarw.9:1) to give diacetate 23 as a yellowish foam (6.0 g, 9.1 mmol, 85%). Compound 23 had R.sub.f 0.35 (Tol/EtOAc 10:1). .sup.1H NMR (CDCl.sub.3) 7.29-7.17 (m, 2H, H.sub.Ar), 7.06-7.00 (m, 3H, H.sub.Ar), 5.81-5.72 (m, 1H, CH.sub.All), 5.50 (dd.sub.po, 1H, H-4), 5.37 (d, 1H, J=7.3 Hz, H-1), 5.22-5.17 (m, 1H, CH.sub.2All), 5.13-5.09 (m, 1H, CH.sub.2All), 4.61 (d, 1H, J=12.4 Hz, CH.sub.2Bn), 4.53 (dd, 1H, Hz, J.sub.2,3=11.2 Hz, H-2), 4.44 (dd, 1H, J.sub.3,4=4.3 Hz, H-3), 4.39 (dd, 1H, J.sub.5,6a=6.6 Hz, J.sub.6a,6b=11.7 Hz, H-6a), 4.37 (dd, 1H, J.sub.5,6b=5.7 Hz, H-6b), 4.30 (dt.sub.po, 1H, J.sub.4,5=3.1 Hz, H-5), 4.23-4.18 (m, 1H, CH.sub.2All), 4.20 (d.sub.po, 1H, CH.sub.2Bn), 4.01-3.96 (m, 1H, CH.sub.2All), 2.21 (s, 3H, CH.sub.3Ac), 2.17 (s, 3H, CH.sub.3Ac). .sup.13C NMR (CDCl.sub.3) 170.0 (CO.sub.Ac), 162.9 (CO.sub.NTCP), 139.9, 137.4, 129.6, 127.0, 125.2 (C.sub.q,Ar), 133.4 (CH.sub.All), 129.0, 128.2, 128.1, 127.6 (C.sub.Ar), 117.7 (CH.sub.2All), 95.4 (C-1, .sup.1J.sub.C,H=169 Hz), 72.7 (C-5), 72.1 (CH.sub.2Bn), 71.1 (C-3), 69.3 (CH.sub.2All), 68.8 (C-4), 62.9 (C-6), 53.2 (C-2). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.28H.sub.29Cl.sub.4N.sub.2O.sub.9, 677.0627; found 677.0622.

    [0516] 4,6-Di-O-acetyl-3-O-benzyl-2-deoxy-2-tetrachlorophthalimido-/-L-altropyranose (24). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (59 mg, 0.07 mmol, 0.02 equiv.) was stirred in anhyd. THF (5.0 mL) under an H.sub.2 atmosphere at rt for 30 min. The resulting yellow solution was degassed several times with Ar and transferred by use of a cannula into a solution of allyl glycoside 23 (2.3 g, 3.4 mmol, 1.0 equiv.) in anhyd. THF (25 mL). After stirring at rt for 1 h, NIS (864 mg, 3.8 mmol, 1.05 equiv.) and H.sub.2O (5 mL) were added. After 2 h, a TLC analysis (Tol/EtOAc 4:1) revealed the presence of a compound (R.sub.f 0.35) more polar that allyl glycoside 23 (R.sub.f 0.65). 10% Aq. Na.sub.2SO.sub.3 was added and the reaction mixture was concentrated to remove the THF and the aq. phase was extracted with DCM (50 mL) thrice. The combined DCM phases were washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography using Tol/EtOAc (8:1.fwdarw.6:1) to give the expected 24 (2.0 g, 3.2 mmol, 92%) as a white floppy solid (/5:1). Hemiacetal 24 ( anomer) had .sup.1H NMR (extracted, CDCl.sub.3) 7.28-7.01 (m, 5H, H.sub.Ar), 5.61 (t, 1H, J.sub.1,2=J.sub.1,OH=7.6 Hz, H-1), 5.60 (dd, 1H, H-4), 4.60 (d, 1H, J=12.4 Hz, CH.sub.2Bn), 4.53 (dd, 1H, J.sub.3,4=3.8 Hz, J.sub.2,3=11.1 Hz, H-3), 4.43-4.35 (m, 3H, H-2, H-6a, H-6b), 4.32 (ddd, 1H, J.sub.4,5=2.1 Hz, J.sub.5,6=5.1 Hz, J.sub.5,6=7.1 Hz, H-5), 4.22 (d, 1H, CH.sub.2Bn), 3.49 (d, 1H, OH). .sup.13C NMR (CDCl.sub.3) 170.6, 170.4 (CO.sub.Ac), 163.1 (CO.sub.NTCP), 140.0, 137.8, 137.4 (C.sub.q,Ar), 129.7, 129.0, 128.1 (2C), 127.7 (C.sub.Ar), 128.3, 128.2, 127.8, 125.3, 90.4 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 73.8 (C-3), 71.9 (CH.sub.2Bn), 70.8 (C-5), 66.9 (C-4), 62.6 (C-6), 55.0 (C-2), 21.0, 20.8 (2C, CH.sub.3Ac). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.25H.sub.25C.sub.14N.sub.2O.sub.9, 637.0314; found 637.0336.

    [0517] Hemiacetal 24 ( anomer) had .sup.1H NMR (extracted, CDCl.sub.3) 7.28-7.01 (m, 5H, H.sub.Ar), 5.64 (dd, 1H, J.sub.3,4=2.0 Hz, J.sub.4,5=2.8 Hz, H-4), 5.43 (t, 1H, J.sub.1,2=J.sub.1,OH=4.0 Hz, H-1), 5.21 (dd, 1H, J.sub.5,6a=3.2 Hz, J.sub.5,6b=11.2 Hz, H-5), 4.75 (dd, 1H, J.sub.1,2=3.2 Hz, J.sub.2,3=10.4 Hz, H-2), 4.69 (d, 1H, J=10.8 Hz, CH.sub.2Bn), 4.60 (d, 1H, J=12.4 Hz, CH.sub.2Bn), 4.54-4.49 (m.sub.o, 2H, H-6a, H-6b), 4.43-4.35 (m.sub.o, 1H, CH.sub.2Bn,), 4.21-4.17 (m.sub.po, 1H, H-3), 4.11 (d, 1H, OH). .sup.13C NMR (CDCl.sub.3), 170.9, 170.3 (CO.sub.Ac), 163.8 (CO.sub.NTCP), 140.2, 137.8 (C.sub.q,Ar), 128.3, 128.2, 127.8, 125.2 (C.sub.Ar), 93.1 (C-1.sub.A, .sup.1J.sub.C,H=175 Hz), 75.2 (C-3), 71.5 (CH.sub.2Bn), 67.6 (C-5), 67.5 (C-4), 64.5 (C-6), 53.0 (C-2), 21.4, 21.0 (2C, CH.sub.3Ac). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.25H.sub.25C.sub.14N.sub.2O.sub.9, 637.0314; found 637.0336.

    [0518] 4,6-Di-O-acetyl-3-O-benzyl-2-deoxy-2-tetrachlorophthalimido-/-L-altropyranosyl (N-phenyl)trifluoroacetimidate (25). Hemiacetal 24 (1.5 g, 2.4 mmol, 1.0 equiv.) was dissolved in acetone (20 mL). PTFACl (580 L, 3.6 mmol, 1.5 equiv.) was added followed by the addition of cesium carbonate (947 mg, 2.9 mmol, 1.2 equiv.). The reaction mixture was stirred at rt. After 2 h, a TLC analysis (Tol/EtOAc 6:1) showed the presence of a new compound (R.sub.f 0.65) and the absence of hemiacetal 24 (R.sub.f 0.2). The reaction mixture was filtered through a pad of Celite and solids were washed with acetone (10 mL) twice. The filtrate was concentrated and the residue was purified by flash chromatography (cHex/EtOAc 9:1) to give PTFA 25 as a white solid (1.8 g, 2.2 mmol, 94%). Donor 25 (a/P 3:2) had R.sub.f 0.65 (cHex/EtOAc 9:1). .sup.1H NMR (CDCl.sub.3) 7.29-6.99, 6.72, 6.54 (m, 14H, H.sub.Ar), 6.54, (bs.sub.o, 0.6H, H-1.sub.), 6.43 (brs, 0.4H, H-1.sub.), 5.71 (d, 0.4H, J.sub.4,5=2.4 Hz, H-4), 5.54 (dd, 0.6H, J.sub.3,4=3.2 Hz, J.sub.4,5=3.6 Hz, H-4a), 5.45 (dd, 0.4H, J.sub.3,4=2.8 Hz, J.sub.2,3=9.6 Hz, H-3), 4.97 (dd, 0.4H, J.sub.1,2=3.6 Hz, H-2), 4.71 (d, 0.4H, J=10.4 Hz, CH.sub.2Bn), 4.7 (dd, 0.6H, J.sub.1,2=4.8 Hz, J.sub.2,3=10.8 Hz, H-2), 4.65 (d, 0.6H, J=12.4 Hz, CH.sub.2Bn), 4.60 (dd.sub.po, 0.4H, J.sub.6a,6b=8.0 Hz, J.sub.5,6b=4.4 Hz, H-6a.sub.), 4.65 (d, 0.6H, CH.sub.2Bn), 4.51-4.35 (m, 2.2H, H-5.sub., H-5.sub., H-6a.sub., H-6 b.sub., H-3.sub.), 4.33 (dd, 0.6H, J.sub.6a,6b=12.4 Hz, J.sub.5,6b=4.8 Hz, H-6b.sub.), 4.23 (d, 0.6H, CH.sub.2Bn), 2.23, 2.16 (2s, 1.8H, CH.sub.3Ac), 2.21, 2.17 (2s, 1.2H, CH.sub.3Ac). .sup.13C NMR (CDCl.sub.3) 170.4 (2C, CO.sub.Ac,), 170.3, 169.9 (2C, CO.sub.Ac,), 163.2 (CO.sub.TCP,), 162.6 (CO.sub.TCP,), 142.9, 142.5, 140.3, 137.8, 129.8, 126.9, 124.5 (C.sub.Ar, , q), 140.1, 137.2, 129.7, 126.9, 124.5 (C.sub.Ar, , q), 129.0, 128.6 (2C), 128.3, 128.2, 128.1, 127.9, 127.7, 125.2, 124.5 (2C), 119.4, 119.1 (C.sub.Ar, , C.sub.Ar, ), 95.2 (C-1.sub.A, .sup.1J.sub.C,H=182 Hz, C-1.sub., .sup.1J.sub.C,H=177 Hz), 76.3 (C-5.sub.), 74.2 (C-5.sub.), 72.3 (CH.sub.2Bn), 71.8 (CH.sub.2Bn), 70.8 (C-3.sub.), 67.5 (C-4.sub., C-4.sub.), 66.7 (C-3.sub.), 63.3 (C-6.sub.), 63.2 (C-6.sub.), 52.3 (C-2.sub.), 51.5 (C-2.sub.), 21.4, 20.9 (CH.sub.3Ac), 20.8, 20.6 (CH.sub.3Ac).

    [0519] Allyl 4,6-di-O-acetyl-3-O-benzyl-2-deoxy-2-tetrachlorophthalimido--L-altropyranosyl-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (26). A mix of acceptor 8 (2.03 g, 5.45 mmol, 1.0 equiv.) and PTFA 25 (5.4 g, 6.54 mmol, 1.2 equiv.) was coevaporated with anhyd. toluene (20 mL), dried under high vacuum for 1 h, and then dissolved in anhyd. DCE (130 mL). Freshly activated MS 4 (1.0 g) was added and after stirring for 45 min at rt under an Ar atmosphere, the reaction mixture was cooled to 0 C. and TMSOTf (59 L, 327 mmol, 0.05 equiv.) was added. After 20 min at 0 C., a TLC follow up (Tol/EtOAc 3:1) showed a new spot (R.sub.f 0.4) and no remaining acceptor 8 (R.sub.f 0.35). Et.sub.3N was added and the suspension was passed through a fitted funnel. Solids were washed DCM (30 mL) twice and the filtrate was concentrated under reduced pressure. Flash chromatography using Tol/EtOAc (8:1.fwdarw.6:1) yielded the desired disaccharide 26 (4.7 g, 4.83 mmol, 88%) as a white solid. Diacetate 26 had .sup.1H NMR (CDCl.sub.3) 7.28-7.16 (m, 2H, H.sub.Ar), 7.00 (brs, 3H, H.sub.Ar), 6.67 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B), 5.83-5.74 (m, 1H, CH.sub.All), 5.56 (d, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A), 5.46 (dd, 1H, J.sub.4,5=2.8 Hz, H-4.sub.A), 5.22-5.16 (m, 1H, CH.sub.2All), 5.17-5.10 (m, 1H, CH.sub.2All), 4.75 (d, 1H, J.sub.1,2=8.3 Hz, H-1.sub.B), 4.58 (dd.sub.po, 1H, J.sub.2,3=10.8 Hz, J.sub.3,4=3.6 Hz, H-3.sub.B), 4.57 (do, 1H, CH.sub.2Bn), 4.57 (dd.sub.po, 1H, J.sub.2,3=11.2 Hz, H-2.sub.A), 4.41 (dd.sub.po, 1H, J.sub.3,4=3.9 Hz, H-3.sub.A), 4.40-4.35 (m, 3H, H-5.sub.A, H-6a.sub.A, H-6b.sub.A), 4.29-4.24 (m, 1H, CH.sub.2All), 4.16 (d, 1H, J=12.4 Hz, CH.sub.2Bn), 4.00-3.95 (m, 1H, H-4.sub.B, CH.sub.2All), 3.68 (dq, J.sub.4,5=1.0 Hz, H-5.sub.B), 3.55 (ddd, 1H, J.sub.2,3=11.0 Hz, H-2.sub.B), 2.18 (s, 3H, CH.sub.3Ac), 2.17 (s, 3H, CH.sub.3Ac), 1.37 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 170.5, 170.3 (CO.sub.Ac), 162.9 (CO.sub.NHTCA), 161.5 (CO.sub.NTCP), 139.9, 137.8, 137.2 (C.sub.q, Ar), 133.4 (CH.sub.All), 129.0, 128.2, 128.1, 128.0, 127.6, 125.2 (C.sub.Ar), 118.0 (CH.sub.2All), 97.9 (C-1.sub.A, .sup.1J.sub.C,H=171 Hz), 97.5 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.2 (CCl.sub.3), 76.6 (C-3.sub.B), 73.2 (C-3.sub.A), 72.1 (CH.sub.2Bn), 70.6 (C-5.sub.A), 70.1 (CH.sub.2All), 69.0 (C-5.sub.B), 66.5 (C-4.sub.A), 65.4 (C-4.sub.B), 62.8 (C-6.sub.A), 55.3 (C-2.sub.B), 53.1 (C-2.sub.A), 20.8 (2C, CH.sub.3Ac), 17.5 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.36H.sub.38C.sub.17N.sub.6O.sub.12, 991.0367; found 991.0355.

    [0520] Allyl (2-acetamido-4,6-di-O-acetyl-3-O-benzyl-2-deoxy--L-altropyranosyl)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (27). Ethylenediamine (41 L, 617 mol, 4.0 equiv.) was added to disaccharide 24 (150 mg, 154 mol, 1.0 equiv.) in n-butanol (8 mL) and the solution was heated at 70 C. for 72 h. A TLC follow up (Tol/EtOAc 4:1) indicated the absence of phtalimide 24 and after the reaction mixture reached rt, volatiles were eliminated and the residue was coevaporated with toluene (5 mL) twice. Acetic anhydride (0.15 mL, 1.5 mmol, 10 equiv.) was added to the residue in pyridine (4 mL) and the system was stirred at rt. In the absence of further evolution after for 4 h (TLC:Tol/EtOAc 4:1), volatiles were evaporated and the residue was purified by column chromatography eluting with Tol/EtOAc (80:20.fwdarw.70:30) to give 27 (76 mg, 101 mol, 65%) as a white solid. The desired 27 had R.sub.f 0.45 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.41-7.30 (m, 5H, H.sub.Ar), 6.67 (d, 1H, J.sub.2,NH=8.3 Hz, NH.sub.B), 5.83-5.74 (m, 1H, CH.sub.All), 5.56 (d, 1H, J.sub.1,2=7.6 Hz, NH.sub.A), 5.29-5.24 (m, 1H, CH.sub.2All), 5.20-5.17 (m, 1H, CH.sub.2All), 5.02 (dd, 1H, J.sub.3,4=3.4 Hz, J.sub.4,5=8.2 Hz, H-4.sub.A), 4.90 (d, 1H, J.sub.1,2=2.2 Hz, H-1.sub.A), 4.87 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.67 (d, 1H, J=12.3 Hz, CH.sub.2Bn), 4.64 (d, 1H, CH.sub.2Bn), 4.59 (ddd.sub.po, 1H, J.sub.5,6a=5.6 Hz, H-5.sub.A), 4.55 (dd.sub.po, 1H, J.sub.3,4=3.5 Hz, H-3.sub.B), 4.37-4.31 (m.sub.po, 1H, CH.sub.2All), 4.44-4.34 (m, 2H, H-2.sub.A, H-6a.sub.A), 4.18 (dd, 1H, J.sub.5,6b=2.9 Hz, J.sub.6a,6b=12.0 Hz, H-6b.sub.A), 4.09-4.04 (m, 1H, CH.sub.2All), 3.91 (dd, 1H, J.sub.2,3=5.3 Hz, H-3.sub.A), 3.84 (brd, 1H, J.sub.3,4=3.4 Hz, H-4.sub.B), 3.77 (brq, J.sub.4,5=1.0 Hz, H-5.sub.B), 3.58 (ddd, 1H, J.sub.2,3=10.8 Hz, H-2.sub.B), 2.18, (s, 3H, CH.sub.3Ac), 2.17 (s, 3H, CH.sub.3Ac), 1.37 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 170.5, 170.3 (2C, CO.sub.Ac), 162.9 (CO.sub.NHTCA), 161.5 (CO.sub.NTCP), 139.9, 137.8, 137.2, (C.sub.q, Ar), 133.4 (CH.sub.All), 129.0, 128.2, 128.1, 128.0, 127.6, 125.2 (C.sub.Ar), 118.0 (CH.sub.2All), 97.9 (C-1.sub.A, .sup.1J.sub.C,H=171 Hz), 97.5 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.2 (CCl.sub.3), 76.6 (C-3.sub.B), 73.2 (C-3.sub.A), 72.1 (CH.sub.2Bn), 70.6 (C-5.sub.A), 70.1 (CH.sub.2All), 69.0 (C-5.sub.B), 66.5 (C-4.sub.A), 65.4 (C-4.sub.B), 62.8 (C-6.sub.A), 55.3 (C-2.sub.B), 53.1 (C-2.sub.A), 20.8 (2C, CH.sub.3Ac), 17.5 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.30H.sub.39Cl.sub.3N.sub.5O.sub.11, 750.1711; found 750.1746.

    [0521] Allyl 2-acetamido-3-O-benzyl-2-deoxy--L-altropyranosyl-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (28). NaOMe (26 L, 25% NaOMe in MeOH, 0.2 equiv.) was added to disaccharide 26 (500 mg, 514 mol, 1.0 equiv.) in anhyd. methanol (15 mL). After stirring at rt for 1 h, a TLC follow up (Tol/EtOAc 1:2) indicated reaction completion and Dowex resin (H.sup.+) was added pinch by pinch to reach pH 7. The suspension was filtered by passing through a fitted funnel and the filtrate was concentrated and dried under high vacuum. The obtained diol had (HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.32H.sub.34Cl.sub.7N.sub.6O.sub.10, 907.0156; found 907.0156) Following extensive drying, the intermediate diol was dissolved in THF/MeOH (1:4, 15 mL). Ethylenediamine (137 L, 2.0 mmol, 4.0 equiv.) was added and the mixture was heated at 70 C. for 48 h. At reaction completion (TLC:EtOAc), the reaction mixture was cooled down, concentrated under reduced pressure, and coevaporated with toluene (5 mL) twice. Et.sub.3N (500 L) and acetic anhydride (485 L, 5.1 mmol, 10 equiv.) were added to the residue in methanol (10 mL). After stirring at rt for 2 h, a TLC follow up (EtOAc/MeOH 9:1) showed reaction completion. Volatiles were evaporated and the residue was purified by column chromatography (DCM/MeOH 95:5) to give the desired 28 (235 mg, 353 mol, 69%) as a white solid. Diol 28 had R.sub.f 0.2 (EtOAc). .sup.1H NMR (DMSO-d.sub.6) 8.85 (d, 1H, J=9.0 Hz, NH.sub.B), 7.93 (d.sub.po, 1H, J=8.7 Hz, NH.sub.A), 7.44-7.22 (m, 5H, H.sub.Ar), 5.86-5.76 (m, 1H, CH.sub.All), 5.26-5.21 (m, 1H, CH.sub.2All), 5.12-5.09 (m, 1H, CH.sub.2All), 4.79 (d, 1H, J.sub.1,2=1.7 Hz, H-1.sub.A), 4.65 (d, 1H, J=12.0 Hz, CH.sub.2Bn), 4.61-4.56 (m, 2H, OH), 4.51 (d.sub.po, 1H, J.sub.1,2=8.3 Hz, H-1.sub.B), 4.50 (d.sub.po, 1H, CH.sub.2Bn), 4.27 (ddd, 1H, J.sub.2,3=3.9 Hz, H-2.sub.A), 4.22-4.17 (m, 1H, CH.sub.2All), 4.11-4.07 (m.sub.po, 1H, J.sub.3,4=3.6 Hz, H-3.sub.B), 4.09 (brs.sub.o, 1H, H-4.sub.B), 4.04 (ddd, 1H, J.sub.4,5=9.2 Hz, J.sub.5,6a=2.1 Hz, J.sub.5,6b=7.1 Hz, H-5.sub.A), 3.99-3.96 (m, 1H, CH.sub.2All), 3.90-3.84 (m, 1H, H-2.sub.B), 3.76-3.70 (m, 2H, H-5.sub.B, H-6a.sub.A), 3.64 (m, 1H, H-4.sub.A), 3.54-3.48 (m.sub.po, 1H, H-6b.sub.A), 3.47 (pt, 1H, J.sub.3,4=4.1 Hz, H-3.sub.A), 1.79 (s, 3H, CH.sub.3Ac), 1.27 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13CNMR (DMSO-d.sub.6) 169.0 (CO.sub.NHTCA), 162.2 (CO.sub.NHAc), 139.4 (C.sub.q,Ar), 134.8 (CH.sub.All), 128.2, 127.9, 127.4 (C.sub.Ar), 116.8 (CH.sub.2All), 102.0 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 100.3 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 93.5 (CCl.sub.3), 77.4 (C-3.sub.B), 76.9 (C-3.sub.A), 71.6 (C-5.sub.A), 71.0 (CH.sub.2Bn), 69.7 (C-5.sub.B), 69.3 (CH.sub.2All), 65.3 (C-4.sub.B), 64.9 (C-4.sub.A), 62.0 (C-6.sub.A), 53.4 (C-2.sub.B), 49.4 (C-2.sub.A), 22.9 (CH.sub.3Ac), 17.7 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.26H.sub.35Cl.sub.3N.sub.5O.sub.9, 666.1486; found 666.1500.

    [0522] Allyl (benzyl 2-acetamido-3-O-benzyl-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (29). TEMPO (89 mg, 0.057 mmol, 0.2 equiv.) and BAIB (242 mg, 0.75 mmol, 2.5 equiv.) were added to diol 28 (200 mg, 301 mol, 1.0 equiv.) in DCM/H.sub.2O (2:1, 15 mL). The reaction was stirred at rt for 4 h. At completion as indicated by TLC (EtOAc/MeOH 9:1), 10% aq. Na.sub.2SO.sub.3 was added and the biphasic mixture was diluted with DCM (20 mL). The aq. phase was separated and extracted with DCM (10 mL) twice. The aq. phase was acidified with dilute aq. HCl to reach pH 1 and washed with DCM (10 mL) thrice. The combined organic phases were washed with brine, dried by passing through a phase separator filter and concentrated under reduced pressure. Benzyl bromide (142 L, 1.2 mmol, 4.0 equiv.) and K.sub.2CO.sub.3 (83 mg, 0.60 mmol, 2.0 equiv.) were added to the residue in anhyd. DMF (2 mL). After stirring at rt for 1 h, the reaction mixture was diluted with H.sub.2O (50 mL) and the aq. layer was extracted with DCM (20 mL) thrice. The organic phases were combined, washed with brine (20 mL), dried over anhyd. Na.sub.2SO.sub.4, filtered and concentrated in vacuo. Flash chromatography eluting with Tol/EtOAc (3:1.fwdarw.2:1) gave the benzyl ester 29 (85 mg, 110 mol, 36%) as a white solid. Disaccharide 29 had R.sub.f 0.3 (Tol/EtOAc 1:1). .sup.1H NMR (CDCl.sub.3) 7.43-7.31 (m, 10H, H.sub.Ar), 6.81 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.89-5.79 (m, 1H, CH.sub.All), 7.93 (d, 1H, J.sub.2,NH=8.4 Hz, NH.sub.A), 5.28 (d, 1H, J.sub.2,NH=12.1 Hz, CH.sub.2Bn-6, CH.sub.2All), 5.29-5.23 (m.sub.po, 1H, CH.sub.2All), 5.22 (d.sub.po, 1H, CH.sub.2Bn-6), 5.20-5.16 (m, 1H, CH.sub.2All), 4.93 (d, 1H, J.sub.1,2=3.2 Hz, H-1.sub.A), 4.78 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.73 (d, 1H, J=11.8 Hz, CH.sub.2Bn), 4.69 (d.sub.po, 1H, J.sub.4,5=7.8 Hz, H-5.sub.A), 4.66 (d, 1H, CH.sub.2Bn), 4.51 (dd, 1H, J.sub.3,4=3.7 Hz, J.sub.2,3=10.7 Hz, H-3.sub.B), 4.34-4.39 (m, 2H, H-2.sub.A, CH.sub.2All), 4.12 (ddd, 1H, J.sub.4,5=7.8 Hz, H-4.sub.A), 4.07-4.01 (m, 1H, CH.sub.2All), 3.85 (dd, 1H, J.sub.2,3=5.0 Hz, J.sub.3,4=3.5 Hz, H-3.sub.A), 3.80 (d, 1H, J.sub.3,4=3.2 Hz, H-4.sub.B), 3.61 (dq, 1H, J.sub.4,5=0.9 Hz, H-5.sub.B), 3.55 (ddd, J.sub.2,3=10.8 Hz, 1H, H-2.sub.B), 2.76 (d, 1H, J.sub.4,OH=8.2 Hz, OH), 1.95 (s, 3H, CH.sub.3Ac), 1.27 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 169.5 (CO.sub.NHTCA), 169.4 (C-6), 162.1 (CO.sub.NHAc), 137.6, 135.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 128.7, 128.6 (2C), 128.4 (2C), 128.2, 128.0 (C.sub.Ar), 117.9 (CH.sub.2All), 100.6 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.2 (CCl.sub.3), 76.7 (C-3.sub.B), 75.2 (C-3.sub.A), 72.0 (CH.sub.2Bn), 71.0 (C-5.sub.A), 70.0 (CH.sub.2All), 69.6 (C-5.sub.B), 67.4 (CH.sub.2Bn-6), 65.6 (C-4.sub.A), 65.4 (C-4.sub.B), 55.3 (C-2.sub.B), 49.5 (C-2.sub.A), 23.2 (CH.sub.3Ac), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.33H.sub.38Cl.sub.3N.sub.5O.sub.10Na, 792.1582; found 792.1584.

    [0523] AB Building Block from an A Donor Whereby the Protecting Groups at the 4.sub.A-OH 6.sub.A-OH are Orthogonal to Each Other

    ##STR00089##

    [0524] Allyl 2-azido-3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranoside (30). CSA (4.1 g, 17.7 mmol, 0.5 equiv.) was added to acetal 12 (15.0 g, 35.4 mmol, 1.0 equiv.) in MeOH/DCM (4:1, 170 mL). After stirring at rt for 2 h, a TLC follow up (Tol/EtOAc 4:1) indicated reaction completion as shown by the absence of the starting 12 (R.sub.f 0.65) and the presence of a non migrating spot. 5% Aq. NaHCO.sub.3 (300 mL) was added followed by EtOAc (500 mL). The organic phase was separated and washed with brine (500 mL). The organic phase was dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude product was dried under high vacuum. tert-Butyldiphenylchlorosilane (10.1 mL, 38.9 mmol, 1.1 equiv.) and imidazole (3.1 g, 46.0 mmol, 1.3 equiv.) were added to the crude diol in anhyd. DMF (180 mL) at 0 C. The reaction mixture was allowed to reach rt slowly and stirred overnight at this temperature. Methanol (10 mL) was added and after 30 min, volatiles were evaporated under reduced pressure. The residue was dissolved in EtOAc (500 mL) and the organic layer was washed with 90% aq. brine (500 mL), separated, dried over Na.sub.2SO.sub.4, and concentrated. 2-(Bromomethyl)naphthalene (10.9 g, 49.6 mmol, 1.4 equiv.) was added to the crude intermediate in DMF (230 mL). The solution was cooled to 0 C. and NaH (60% in mineral oil, 1.7 g, 70.8 mmol, 2.0 equiv.) was added portion wise. After stirring vigorously for 2 h while the bath temperature slowly reached rt, a TLC follow up indicated reaction completion. The reaction mixture was diluted with DCM (1 L) and 5% aq. NH.sub.4Cl (500 mL) was added. The organic layer was washed with water (1.5 L) and brine (1 L), dried over Na.sub.2SO.sub.4 and concentrated. The crude product was purified by flash chromatography (cHex/EtOAc 12:1.fwdarw.10:1) to give the fully protected 30 (21.6 g, 30.2 mmol, 85%) as a light yellow oil. Allyl glycoside 30 had R.sub.f 0.8 (Tol/EtOAc 10:1). .sup.1H NMR (CDCl.sub.3) 7.84-7.30 (m, 22H, H.sub.Ar), 5.98-5.88 (m, 1H, CH.sub.All), 5.34-5.28 (m, 1H, CH.sub.2All), 5.21-5.17 (m, 1H, CH.sub.2All), 4.79 (d, 1H, J=12.2 Hz, CH.sub.2Nap), 4.79 (d, 1H, CH.sub.2Nap), 4.73 (d, 1H, J.sub.1,2=4.7 Hz, H-1), 4.67 (d, 1H, J=11.9 Hz, CH.sub.2Bn), 4.61 (d, 1H, CH.sub.2Bn), 4.31-4.25 (m, 1H, CH.sub.2All), 4.19 (pq, 1H, H-5), 4.07-4.01 (m, 1H, CH.sub.2All), 3.98-3.94 3.97 (dd.sub.po, 1H, H-2), 3.95 (dd.sub.po, 1H, J.sub.4,5=5.3 Hz, H-4), 3.77 (brd, 2H, J.sub.5,6a=4.5 Hz, J.sub.5,6b=4.5 Hz, H-6a, H-6b), 3.74 (dd.sub.po, 1H, J.sub.3,4=3.5 Hz, J.sub.2,3=8.0 Hz, H-3), 1.00 (s, 9H, CH.sub.3, TBDPS). .sup.13C NMR (CDCl.sub.3), 137.8, 135.5, 133.2, 133.0 (C.sub.q, Ar), 133.8 (CH.sub.All), 135.6, 135.6, 129.7, 128.3, 128.1, 127.9, 127.7 (2C), 126.6 (2C), 126.0, 125.9, 125.8 (C.sub.Ar), 117.2 (CH.sub.2All), 98.7 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 76.2 (C-3), 72.9 (C-5), 72.3 (CH.sub.2Nap, CH.sub.2Bn), 72.1 (C-4), 68.7 (CH.sub.2All), 63.7 (C-6), 61.8 (C-2), 26.9, 26.7 (3C, CH.sub.3,TBDPS), 19.1 (C.sub.TBDPS). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.34H.sub.47N.sub.3O.sub.5SiNa, 736.3783; found 736.3177.

    [0525] Allyl 3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-(2-naphthylmethyl)-2-tetrachlorophthalimido--L-altropyranoside (31). Zn dust (8.2 g, 126 mmol, 10.0 equiv.) and AcOH (7.2 mL, 126 mmol, 10.0 equiv.) were added to azide 30 (9.0 g, 12.6 mmol, 1.0 equiv.) in anhyd. THF (85 mL). After stirring for 1 h, a TLC analysis (Tol/EtOAc 10:1) showed the absence of azide 30 (R.sub.f 0.8) and the presence of a more polar spot (R.sub.f 0.0). The suspension was filtered over a pad of Celite and washed with DCM. The DCM layer was washed with satd aq. NaHCO.sub.3, water, and brine, dried over Na.sub.2SO.sub.4, concentrated under reduced pressure, and dried under high vacuum. The crude amine was dissolved in DCM and tetrachlorophthalic anhydride (2.2 g, 7.5 mmol, 0.6 equiv.) was added. The mixture was stirred at rt for 30 min. Et.sub.3N (2.1 mL, 15.1 mmol, 1.2 equiv.) was added followed by more tetrachlorophthalic anhydride (2.2 g, 7.5 mmol, 0.6 equiv.). The reaction was stirred for another 30 min at rt, at which time a TLC follow up (EtOAc) indicated reaction completion. Volatiles were evaporated and dried under high vacuum. The crude was dissolved in pyridine (60 mL) and Ac.sub.2O (5.9 mL, 63.0 mmol, 5.0 equiv.) was added. After heating to 80 C. for 10 min, a TLC analysis (cHex/EtOAc 9:1) showed full consumption of the intermediate and the presence of a less polar spot. At completion, the mixture was concentrated under reduced pressure and coevaporated with toluene (30 mL) twice. The crude was taken in DCM (300 mL) and the DCM layer was washed with water (300 mL) and brine (300 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (cHex/EtOAc 93:7.fwdarw.88:12) to give the fully protected 31 (9.54 g, 10.4 mmol, 82%) as a yellowish dense oil. Allyl glycoside 31 had R.sub.f 0.7 (cHex/EtOAc, 10:1). .sup.1H NMR (CDCl.sub.3) 7.83-7.35 (m, 18H, H.sub.Ar), 7.04 (brs, 4H, H.sub.Ar), 5.83-5.73 (m, 1H, CH.sub.All), 5.27 (d, 1H, J.sub.1,2=7.0 Hz, H-1), 5.19-5.14 (m, 1H, CH.sub.2All), 5.08-5.05 (m, 1H, CH.sub.2All), 4.94 (d, 1H, J=12.7 Hz, CH.sub.2Nap), 4.85 (d.sub.po, 1H, CH.sub.2Nap), 4.85 (dd.sub.po, 1H, J.sub.2,3=11.2 Hz, H-2), 4.60 (d, 1H, J=12.3 Hz, CH.sub.2Bn), 4.3.6 (dd, 1H, J.sub.3,4=3.6 Hz, H-3), 4.30 (ddd.sub.po, 1H, J.sub.4,5=3.3 Hz, H-5), 4.25-4.19 (m, 1H, CH.sub.2All), 4.16 (pt, 1H, H-4), 4.10 (d, 1H, CH.sub.2Bn), 3.98-3.93 (m, 1H, CH.sub.2All), 3.83 (brd, 2H, J.sub.5,6a=6.1 Hz, J.sub.5,6b=6.1 Hz, H-6a, H-6b), 1.04 (s, 9H, CH.sub.3,TBDPS). .sup.13C NMR (CDCl.sub.3) 163.2 (CO.sub.NTCP), 157.4, 142.4, 139.7, 138.0, 135.9, 133.2, 133.1, 133.0 (2C), 129.5, 127.7, 127.2 (C.sub.q,Ar), 133.9 (CH.sub.All), 135.6 (2C), 129.8, 128.0, 127.9, 127.8, 127.6, 127.5, 127.3, 126.5, 126.0 (2C), 125.8 (C.sub.Ar), 117.1 (CH.sub.2All), 96.1 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 75.6 (C-5), 74.3 (C-3), 72.7 (CH.sub.2Nap), 72.1 (CH.sub.2Bn), 71.8 (C-4), 68.6 (CH.sub.2All), 63.1 (C-6), 53.4 (C-2), 26.9, 26.7 (3C, CH.sub.3,TBDPS), 19.2 (C.sub.TBDPS). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.51H.sub.51Cl.sub.4N.sub.2O.sub.7Si, 971.2220; found 971.2213.

    [0526] 3-O-Benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-(2-naphthylmethyl)-2-tetrachlorophthalimido-/-L-altropyranose (32). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (115 mg, 0.13 mmol, 0.02 equiv.) was dissolved in anhyd. THF (8.0 mL) and stirred for 30 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly with Ar and transferred by means of a cannula into a solution of allyl glycoside 31 (6.5 g, 6.8 mmol, 1.0 equiv.) in anhyd. THF (60 mL). The reaction mixture was stirred for 1 h at rt, at which time a solution of NIS (1.68 g, 7.5 mmol, 1.1 equiv.) in H.sub.2O (15 mL) was added. After stirring for 1 h at rt, a TLC analysis (cHex/EtOAc 8:1) revealed the full consumption of the isomerization product (R.sub.f 0.65) and the presence of a more polar spot (R.sub.f 0.1). 10% Aq. Na.sub.2SO.sub.3 was added and volatiles were evaporated. The aq. phase was extracted with DCM (200 mL) twice. The organic layers were combined, washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. Purification of the residue by flash chromatography (Tol/EtOAc 10:1.fwdarw.8:1) yielded the expected hemiacetal 32 (5.2 g, 5.6 mmol, 82%) as a white floppy solid. Hemiacetal 32 ((a/P 5:1) had R.sub.f 0.6 (cHex/EtOAc 9:1). The ca anomer had .sup.1H NMR (CDCl.sub.3) 7.84-6.99 (m, 22H, HM), 5.25 (dd, 1H, J.sub.1,OH=9.4 Hz, J.sub.1,2=6.8 Hz, H-1), 4.81 (d, 1H, J=12.4 Hz, CH.sub.2Nap), 4.76 (d, 1H, CH.sub.2Nap), 4.67 (dd, 1H, J.sub.2,3=10.8 Hz, H-2), 4.53 (d, 1H, J=12.4 Hz, CH.sub.2Bn), 4.43-4.33 (dd, 1H, J.sub.3,4=3.0 Hz, H-3), 4.30 (ddd, J.sub.4,5=1.8 Hz, 1H, H-5), 4.19 (dd, 1H, H-4), 4.03 (d, 1H, CH.sub.2Bn), 3.91 (dd, 1H, J.sub.5,6a=5.6 Hz, J.sub.6a,6b=10.6 Hz, H-6a), 3.86 (dd, 1H, J.sub.6a,6b=8.2 Hz, H-6b), 3.06 (d, 1H, OH), 1.03 (s, 9H, CH.sub.3TBDPS). .sup.13C NMR (CDCl.sub.3) 163.2 (CO.sub.NTCP), 139.8, 137.8, 137.9, 137.8, 135.6, 135.5, 135.4, 133.2, 133.1, 133.0, 132.8 (C.sub.q,Ar), 135.6, 135.5, 129.9 (2C), 129.0, 128.2 (2C), 128.1 (2C), 127.9, 127.8 (2C), 127.7, 127.4, 126.8, 126.6, 126.0 (2C), 125.9, 125.2 (C.sub.q,Ar), 91.5 (C-1.sub.A, .sup.1J.sub.C,H=171 Hz), 76.1 (C-3), 73.3 (C-5), 72.7 (CH.sub.2Nap), 71.7 (CH.sub.2Bn), 71.7 (C-4), 62.2 (C-6), 56.1 (C-2), 26.8 (CH.sub.3TBDPS), 21.4 (C.sub.TBDPS). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.48H.sub.47C.sub.14N.sub.2O.sub.7Si, 931.1907; found 931.1880.

    [0527] The anomer had .sup.1H NMR (CDCl.sub.3) 7.84-6.99 (m, 22H, H.sub.Ar), 5.37 (dd, 1H, J.sub.1,2=4.1 Hz, J.sub.1,OH=5.6 Hz, H-1), 4.95 (dd, 1H, J.sub.2,3=10.8 Hz, H-2), 4.95 (dd.sub.po, 1H, J.sub.3,4=2.7 Hz, H-3), 4.90 (d.sub.po, 1H, CH.sub.2Nap), 4.85 (d, 1H, J=12.6 Hz, CH.sub.2Nap), 4.41 (d, 1H, J=11.6 Hz, CH.sub.2Bn), 4.28 (bs.sub.o, 1H, H-4), 4.28-4.25 (m, 1H, H-5), 4.21 (d.sub.po, 1H, CH.sub.2Bn), 3.99 (d.sub.po, 1H, H-6a), 3.93-3.84 (m.sub.o, 1H, H-6b), 3.48 (brs, 1H, OH), 1.08 (s, 9H, CH.sub.3TBDPS). .sup.13C NMR (CDCl.sub.3) 163.9 (CO.sub.NTCP), 139.8-125.3 (C.sub.Ar), 92.6 (C-1.sub.A, .sup.1J.sub.C,H=175 Hz), 77.4 (C-3), 72.2 (CH.sub.2Nap), 72.4 (C-4), 71.4 (CH.sub.2Bn), 64.6 (C-6), 53.6 (C-2), 26.8 (CH.sub.3TBDPS), 21.4 (C.sub.TBDPS). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.48H.sub.47C.sub.14N.sub.2O.sub.7Si, 931.1907; found 931.1880.

    [0528] Allyl 3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-(2-napthylmethyl)-2-tetrachlorophthalimido--L-altropyranosyl-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (34). PTFACl (1.47 mL, 7.1 mmol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (1.9 g, 6.0 mmol, 1.1 equiv.) were added to hemiacetal 32 (5.0 g, 5.4 mmol, 1.0 equiv.) in acetone (40 mL). After stirring for 2 h at rt, the reaction mixture was filtered through a pad of Celite and washed with DCM (50 mL) twice. The filtrate was concentrated under reduced pressure and dried under vacuum to give the crude donor 33 (6.0 g, 5.4 mmol, quant.), which was used as such in the next step. The PTFA donor 33 had R.sub.f 0.85 (Tol/EtOAc 10:1). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.56H.sub.47C.sub.14F.sub.3N.sub.2O.sub.7SiNa, 1107.1757; found 1107.1755.

    [0529] A mix of the crude PTFA donor 33 (6.0 g, 5.4 mmol, 1.1 equiv. theo.) and acceptor 8 (1.83 g, 4.9 mmol, 1.0 equiv.) were co-evaporated with anhyd. toluene (30 mL) and then dried under vacuum. Freshly activated MS 4 (4.0 g) was added to the starting materials in anhyd. DCM (90 mL) and the suspension was stirred for 1 h under an Ar atmosphere at rt. After cooling to 15 C., TMSOTf (49 L, 0.05 equiv.) was added slowly and stirring went on for 40 min during which the bath temperature kept at 15 C. A TLC analysis (Tol/EtOAc 10:1) showed the absence of donor 33 and the presence of a new spot (R.sub.f 0.5) in addition to a slight amount of hemiacetal 32 (R.sub.f 0.4). At completion, Et.sub.3N (80 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (50 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (cHex/EtOAc 10:1.fwdarw.8:1) to give disaccharide 34 as a white solid (6.0 g, 4.7 mmol, 96%). The coupling product 34 had .sup.1H NMR (CDCl.sub.3) 7.85-7.81 (m, 4H, H.sub.Ar), 7.67-7.63 (m, 3H, H.sub.Ar), 7.53-7.16 (m, 10H, H.sub.Ar), 7.04-6.98 (m, 5H, H.sub.Ar), 6.67 (m, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B), 5.85-5.75 (m, 1H, CH.sub.All), 5.43 (d, 1H, J.sub.1,2=7.2 Hz, H-1.sub.A), 5.23-5.17 (m, 1H, CH.sub.2All), 5.14-5.11 (m, 1H, CH.sub.2All), 4.94 (d, 1H, J=12.4 Hz, CH.sub.2Nap), 4.87 (dd.sub.po, 1H, J.sub.2,3=11.1 Hz, H-2.sub.A), 4.81 (d, 1H, CH.sub.2Nap), 4.73 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.61 (d, 1H, J=12.0 Hz, CH.sub.2Bn), 4.50 (dd, 2H, J.sub.3,4=3.5 Hz, J.sub.2,3=10.7 Hz, H-3.sub.B), 4.39 (ddd.sub.po, 1H, J.sub.4,5=3.3 Hz, H-5.sub.A), 4.33 (dd.sub.po, 1H, J.sub.3,4=3.5 Hz, H-3.sub.A), 4.29-4.24 (m, 1H, CH.sub.2All), 4.11 (pt.sub.po, 1H, H-4.sub.A), 4.09 (d.sub.po, 1H, CH.sub.2Bn), 4.00-3.94 (m, 1H, CH.sub.2All), 3.86 (d.sub.po, 1H, J.sub.3,4=3.8 Hz, H-4.sub.B), 3.85 (d.sub.po, 1H, J.sub.5,6a=6.4 Hz, H-6a.sub.A), 3.79 (dd, 1H, J.sub.5,6b=5.8 Hz, J.sub.6a,6b=11.0 Hz, H-6b.sub.A), 3.51 (dd, 1H, H-2.sub.B), 3.47 (dq, J.sub.4,5=1.3 Hz, H-5.sub.B), 1.82 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B), 1.05 (s, 9H, CH.sub.3TBDPS). .sup.13C NMR (CDCl.sub.3) 163.1 (CO.sub.NHTCA), 161.5 (CO.sub.NTCP), 139.8, 137.8, 137.7, 135.7, 133.2, 133.0 (2C), 132.9, 129.6, 127.5 (C.sub.q, Ar), 133.5 (CH.sub.All), 135.6, 135.2, 129.9, 129.0, 128.2, 128.1, 128.0, 127.9, 127.8, 127.7, 127.6, 127.4, 126.5, 126.0, 125.9, 125.2 (C.sub.Ar), 117.8 (CH.sub.2All), 98.5 (C-1.sub.A, .sup.1J.sub.C,H=171 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 92.2 (CCl.sub.3), 75.9 (C-5.sub.A), 75.5 (C-3.sub.B), 73.7 (C-3.sub.A), 72.6 (CH.sub.2Nap), 72.1 (CH.sub.2Bn), 71.4 (C-4.sub.A), 70.0 (CH.sub.2All), 69.2 (C-5.sub.B), 65.4 (C-4.sub.B), 63.1 (C-6.sub.A), 53.3 (C-2.sub.B), 53.1 (C-2.sub.A), 26.9 (CH.sub.3TBDPS), 19.3 (C.sub.TBDPS), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.59H.sub.6OCl.sub.7N.sub.6O.sub.10Si, 1285.1960; found 1285.1948.

    [0530] Allyl 2-acetamido-3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyl-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (35). Ethylenediamine (1.3 mL, 19.3 mmol, 4.0 equiv.) was added to disaccharide 34 (6.2 g, 4.8 mmol, 1.0 equiv.) in THF/MeOH (1:1, 100 mL) at rt and the reaction mixture was stirred at 50 C. for 72 h under an Ar atmosphere. A TLC analysis (Tol/EtOAc 7:3) revealed the absence of the starting 34 (R.sub.f 1.0) and the presence of a new spot (R.sub.f 0.55). The mixture was allowed to reach rt and Et.sub.3N (2.0 mL) was added, followed by acetic anhydride (4.6 mL, 48.9 mmol, 10.0 equiv.). After stirring for 3 h at rt, a TLC analysis (Tol/EtOAc 7:3) showed the presence of a new spot (R.sub.f 0.65) whereas the intermediate amine had been fully consumed. The suspension was filtered by passing through a pad of Celite, washed with DCM (15 mL) thrice and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (cHex/EtOAc 10:1.fwdarw.7:1). Acetamide 35 was obtained as a white solid (4.8 g, 4.6 mmol, 94%). Disaccharide 35 had .sup.1H NMR (CDCl.sub.3) 7.84-7.81 (m, 1H, H.sub.Ar), 7.75-7.62 (m, 7H, H.sub.Ar), 7.50-7.26 (m, 15H, H.sub.Ar), 6.67 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.90-5.80 (m, 1H, CH.sub.All), 5.28-5.23 (m.sub.po, 1H, CH.sub.2All), 5.23 (d.sub.po, 1H, J.sub.2,NH=8.8 Hz, NH.sub.A), 5.19-5.16 (m, 1H, CH.sub.2All), 4.89 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B), 4.78 (d, 1H, J=12.7 Hz, CH.sub.2Nap), 4.74 (d, 1H, CH.sub.2Nap), 4.71 (d.sub.po, 1H, CH.sub.2Bn), 4.69 (brs.sub.po, 1H, J.sub.1,2=1.8 Hz, H-1.sub.A), 4.52 (d, 1H, J=12.1 Hz, CH.sub.2Bn), 4.48 (dd.sub.po, 1H, J.sub.3,4=3.7 Hz, J.sub.2,3=10.9 Hz, H-3.sub.B), 4.47-4.43 (m.sub.o, 2H, H-5.sub.A), 4.41 (ddd, J.sub.2,3=4.3 Hz, H-2.sub.A), 4.34-4.29 (m, 1H, CH.sub.2All), 4.06-4.01 (m.sub.po, 1H, CH.sub.2All), 4.00 (dd.sub.po, 1H, J.sub.5,6a=2.6 Hz, J.sub.6a,6b=11.2 Hz, H-6a.sub.A), 3.95 (dd.sub.po, 1H, J.sub.5,6a=2.6 Hz, J.sub.6a,6b=11.1 Hz, H-6b.sub.A), 3.93 (pt.sub.po, 1H, H-3.sub.A), 3.63 (dd, 1H, J.sub.3,4=3.0 Hz, J.sub.4,5=8.9 Hz, H-4.sub.A), 3.57 (brd, 1H, H-4.sub.B), 3.53 (brq, 1H, H-5.sub.B), 3.41 (ddd, 1H, H-2.sub.B), 1.74 (s, 3H, CH.sub.3NHAc), 1.17 (d, 3H, J.sub.5,6=6.3 Hz, H-6.sub.B), 1.08 (s, 9H, CH.sub.3TBDPS). .sup.13C NMR (CDCl.sub.3) 168.8 (CO.sub.NHTA), 162.1 (CO.sub.NHAc), 138.6, 135.1, 133.5, 133.0 (2C, C.sub.q,Ar), 133.6 (CH.sub.All), 135.7, 135.6, 129.7 (2C), 128.2 (2C), 127.8, 127.7, 127.6, 127.5, 126.8, 126.1, 125.9 (2C, C.sub.Ar), 117.7 (CH.sub.2All), 101.5 (C-1.sub.A, .sup.1J.sub.C,H=168 Hz), 97.5 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 92.2 (CCl.sub.3), 76.0 (C-3.sub.B), 72.5 (C-3.sub.A), 71.5 (CH.sub.2Nap), 70.5 (CH.sub.2Bn), 70.4 (C-4.sub.A), 70.1 (CH.sub.2All), 69.7 (C-5.sub.A), 69.7 (C-5.sub.B), 65.2 (C-4.sub.B), 63.7 (C-6.sub.A), 55.8 (C-2.sub.B), 49.6 (C-2.sub.A), 27.0 (CH.sub.3TBDPS), 23.0 (CH.sub.3NHAc), 19.4 (C.sub.TBDPS), 17.1 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.53H.sub.61Cl.sub.3N.sub.5O.sub.9Si, 1044.3304; found 1044.3325.

    [0531] Allyl 2-acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyl-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (36). TBAF (1.8 g, 5.8 mmol, 1.2 equiv.) was added to disaccharide 35 (4.8 g, 4.8 mmol, 1.0 equiv.) in THF (98 mL) and the reaction mixture was stirred at rt for 4 h. A TLC analysis (Tol/EtOAc 7:3) showed the consumption of the fully protected 35 (R.sub.f 0.65) and the presence of a polar spot. Acetic acid (0.34 mL, 5.8 mmol, 1.2 equiv.) was added and after stirring for 10 min, volatiles were evaporated. The residue was purified by flash chromatography (EtOAc/MeOH 100:0.fwdarw.95:5) to give alcohol 33 (3.2 g, 3.9 mmol, 86%) as a white solid. Disaccharide 36 had R.sub.f 0.15 (EtOAc). .sup.1H NMR (DMSO-d.sub.6) 8.87 (d, 1H, J.sub.2,NH=9.2 Hz, NH.sub.B), 7.93-7.82 (m, 4H, NH.sub.A, H.sub.Ar), 7.75 (brs, 1H, H.sub.Ar), 7.53-7.47 (m, 2H, H.sub.Ar), 7.43-7.39 (m, 3H, H.sub.Ar), 7.32-7.24 (m, 3H, H.sub.Ar), 5.85-5.76 (m, 1H, CH.sub.All), 5.26-5.20 (m, 1H, CH.sub.2All), 5.12-5.09 (m, 1H, CH.sub.2All), 4.81 (d, 1H, J.sub.1,2=1.6 Hz, H-1.sub.A), 4.69 (d.sub.po, 3H, J=11.6 Hz, CH.sub.2Nap), 4.67-4.62 (m, 2H, J=11.8 Hz, OH, CH.sub.2Bn), 4.53 (d.sub.po, 1H, CH.sub.2Bn), 4.51 (d.sub.po, 1H, J.sub.1,2=8.9 Hz, H-1.sub.B), 4.49 (d.sub.po, 1H, CH.sub.2Nap), 4.32 (ddd, 1H, J.sub.2,NH=8.4 Hz, J.sub.2,3=4.5 Hz, H-2.sub.A), 4.24-4.16 (m, 2H, H-5.sub.A, CH.sub.2All), 4.12 (dd.sub.po, 1H, J.sub.3,4=3.6 Hz, J.sub.2,3=10.8 Hz, H-3.sub.B), 4.05 (brd, 1H, H-4.sub.B), 4.00-3.95 (m, 1H, CH.sub.2All), 3.90 (ddd.sub.po, 1H, H-2.sub.B), 3.79 (dd.sub.po, 1H, H-3.sub.A), 3.78-3.74 (m.sub.o, 1H, H-6a.sub.A), 3.72 (bq, 1H, H-5.sub.B), 3.68 (dd, 1H, J.sub.3,4=3.1 Hz, J.sub.4,5=8.9 Hz, H-4.sub.A), 3.53 (ddd, 1H, H-6b.sub.B), 1.74 (s, 3H, CH.sub.3NHAc), 1.24 (s, 9H, CH.sub.3TBDPS). .sup.13C NMR (DMSO-d.sub.6) 169.1 (CO.sub.NHTCA), 162.0 (CO.sub.NHAc), 139.3, 136.4, 133.1, 132.9 (C.sub.q,Ar), 134.8 (CH.sub.All), 128.3, 128.1 (2C), 128.0, 127.6, 126.5, 126.4, 126.3 (C.sub.Ar), 116.8 (CH.sub.2All), 101.9 (C-1.sub.A, .sup.1J.sub.C,H=171 Hz), 100.2 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 93.5 (CCl.sub.3), 77.5 (C-3.sub.B), 73.8 (C-3.sub.A), 72.5 (C-4.sub.A), 70.9 (CH.sub.2Nap), 70.4 (CH.sub.2Bn), 70.2 (C-5.sub.B), 69.6 (C-5.sub.A), 69.3 (CH.sub.2All), 65.3 (C-4.sub.B), 61.6 (C-6.sub.A), 53.4 (C-2.sub.B), 49.4 (C-2.sub.A), 22.9 (CH.sub.3NHAc), 17.6 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.37H.sub.43Cl.sub.3N.sub.5O.sub.9, 806.2126; found 806.2117.

    [0532] Allyl (benzyl 2-acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (37). TEMPO (116 mg, 0.74 mmol, 0.2 equiv.) was added, followed by BAIB (3.0 g, 9.3 mmol, 2.5 equiv.), to a suspension of alcohol 36 (3.0 g, 3.7 mmol, 1.0 equiv.) in DCM/H.sub.2O (2:1, 120 mL). The biphasic mixture stirred vigorously for 2 h at rt, at which point a TLC analysis (EtOAc) revealed the absence of alcohol 36 (R.sub.f 0.15) and the presence of a polar product (R.sub.f 0.0). 10% Aq. Na.sub.2SO.sub.3 was added followed by DCM (80 mL). The DCM layer was separated, and the aq. phase was extracted with DCM (100 mL) twice. The combined organic phases were dried by passing through a phase separator filter and concentrated to dryness. The residue was dissolved in anhyd. DMF (40 mL). Benzyl bromide (1.3 mL, 11.1 mmol, 3.0 equiv.) and K.sub.2CO.sub.3 (670 mg, 4.8 mmol, 1.3 equiv.) were added and the suspension was stirred at rt for 2 h. At completion, satd aq. NH.sub.4Cl was added and the aq. layer was washed with DCM (100 mL) thrice. The organic phases were combined, washed with brine (300 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 7:3.fwdarw.6:4) to give the desired benzyl ester 37 (2.8 g, 3.0 mmol, 85%) as a brown-white solid. Ester 37 had R.sub.f 0.3 (Tol/EtOAc, 4:1). .sup.1H NMR (CDCl.sub.3) 7.84-7.26 (m, 17H, H.sub.Ar), 6.94 (d, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.90-5.80 (m, 1H, CH.sub.All), 5.73 (d, J.sub.2,NH=6.8 Hz, NH.sub.A), 5.33 (d, 1H, J.sub.1,2=5.6 Hz, H-1.sub.A), 5.24-5.20 (m.sub.po, 1H, CH.sub.2All), 5.22 (d.sub.po, 1H, CH.sub.2Bn-6), 5.19-5.15 (m.sub.po, 1H, CH.sub.2All), 5.17 (d, 1H, J=12.0 Hz, CH.sub.2Bn-6), 4.80 (d, 1H, J.sub.4,5=4.6 Hz, H-5.sub.A), 4.77 (d, 1H, J.sub.1,2=8.3 Hz, H-1.sub.B), 4.73 (d, 1H, J=12.7 Hz, CH.sub.2Nap), 4.71 (d, 1H, CH.sub.2Nap), 4.52 (d, 1H, J=12.0 Hz, CH.sub.2Bn), 4.48 (dd.sub.po, 1H, J.sub.3,4=3.8 Hz, H-3.sub.B), 4.48 (d, 1H, CH.sub.2Bn), 4.33-4.28 (m, 1H, CH.sub.2All), 4.10 (dd.sub.po, J.sub.3,4=2.7 Hz, H-4.sub.A), 4.07-3.98 (m, 3H, H-3.sub.A, H-2.sub.A, CH.sub.2All), 3.93 (brd, 1H, J.sub.3,4=3.2 Hz, H-4.sub.B), 3.57 (dt, 1H, J.sub.2,3=10.5 Hz, H-2.sub.B), 3.47 (brq, 1H, H-5.sub.B), 1.86 (s, 3H, CH.sub.3Ac), 1.23 (d, 3H, J.sub.5,6=6.3 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 170.4 (CO.sub.NTCA), 169.3 (C-6.sub.A), 161.9 (CO.sub.NAc), 137.9, 134.9, 134.8, 133.1, 133.0 (C.sub.q, Ar), 133.5 (CH.sub.All), 128.7 (2C), 128.6, 128.3, 128.1, 127.9, 127.8, 127.6, 126.9, 126.1, 126.0, 125.9 (C.sub.Ar), 117.9 (CH.sub.2All), 99.5 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.4 (CCl.sub.3), 76.6 (C-3.sub.B), 73.4 (C-3.sub.A), 72.9 (C-4.sub.A), 71.9 (C-5.sub.A), 71.8 (2C, CH.sub.2Bn, CH.sub.2Nap), 70.1 (CH.sub.2All), 69.3 (C-5.sub.B), 67.5 (CH.sub.2Bn-6), 65.1 (C-4.sub.B), 55.0 (C-2.sub.B), 52.1 (C-2.sub.A), 23.4 (CH.sub.3Ac), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.44H.sub.47Cl.sub.3N.sub.5O.sub.10, 910.2388; found 910.2380.

    [0533] Allyl (benzyl 2-acetamido-3-O-benzyl-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (29). The benzyl ester 37 (see below, 340 mg, 374 mol, 1.0 equiv.) was dissolved in DCM (6.0 mL) and phosphate buffer pH 7 (1.0 mL) was added. The biphasic mixture was cooled to 0 C. and DDQ (144 mg, 636 mol, 1.7 equiv.) was added. Stirring was pursued for 2 h while the bath was allowed to reach rt. The mixture was diluted with DCM (10 mL) and the DCM layer was washed with satd aq. NaHCO.sub.3, water and brine, then dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 65:35.fwdarw.55:45) to give alcohol 29 (290 mg, 325 mol, 87%) as a white solid. Disaccharide 29 had R.sub.f 0.3 (Tol/EtOAc 1:1). .sup.1H NMR (CDCl.sub.3) 7.43-7.31 (m, 10H, H.sub.Ar), 6.81 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.89-5.79 (m, 1H, CH.sub.All), 7.93 (d, 1H, J.sub.2,NH=8.4 Hz, NH.sub.A), 5.28 (d, 1H, J.sub.2,NH=12.1 Hz, CH.sub.2Bn-6, CH.sub.2All), 5.29-5.23 (m.sub.po, 1H, CH.sub.2All), 5.22 (d.sub.po, 1H, CH.sub.2Bn-6), 5.20-5.16 (m, 1H, CH.sub.2All), 4.93 (d, 1H, J.sub.1,2=3.2 Hz, H-1.sub.A), 4.78 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.73 (d, 1H, J=11.8 Hz, CH.sub.2Bn), 4.69 (d.sub.po, 1H, J.sub.4,5=7.8 Hz, H-5.sub.A), 4.66 (d, 1H, CH.sub.2Bn), 4.51 (dd, 1H, J.sub.3,4=3.7 Hz, J.sub.2,3=10.7 Hz, H-3.sub.B), 4.34-4.39 (m, 2H, H-2.sub.A, CH.sub.2All), 4.12 (ddd, 1H, J.sub.4,5=7.8 Hz, H-4.sub.A), 4.07-4.01 (m, 1H, CH.sub.2All), 3.85 (dd, 1H, J.sub.2,3=5.0 Hz, J.sub.3,4=3.5 Hz, H-3.sub.A), 3.80 (d, 1H, J.sub.3,4=3.2 Hz, H-4.sub.B), 3.61 (dq, 1H, J.sub.4,5=0.9 Hz, H-5.sub.B), 3.55 (ddd, J.sub.2,3 10.8 Hz, 1H, H-2.sub.B), 2.76 (d, 1H, J.sub.4,OH=8.2 Hz, OH), 1.95 (s, 3H, CH.sub.3Ac), 1.27 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 169.5 (CO.sub.NHTCA), 169.4 (C-6), 162.1 (CO.sub.NHAc), 137.6, 135.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 128.7, 128.6 (2C), 128.4 (2C), 128.2, 128.0 (C.sub.Ar), 117.9 (CH.sub.2All), 100.6 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.2 (CCl.sub.3), 76.7 (C-3.sub.B), 75.2 (C-3.sub.A), 72.0 (CH.sub.2Bn), 71.0 (C-5.sub.A), 70.0 (CH.sub.2All), 69.6 (C-5.sub.B), 67.4 (CH.sub.2Bn-6), 65.6 (C-4.sub.A), 65.4 (C-4.sub.B), 55.3 (C-2.sub.B), 49.5 (C-2.sub.A), 23.2 (CH.sub.3Ac), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.33H.sub.38Cl.sub.3N.sub.5O.sub.10Na, 792.1582; found 792.1584.

    [0534] (Benzyl 2-acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido-/-D-galactopyranose (38). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (13 mg, 15 mol, 0.02 equiv.) in anhyd. THF (4.0 mL) was degassed repeatedly and stirred for 30 min under a hydrogen atmosphere. The resulting yellow solution was degassed several times with Ar and transferred by means of a cannula into a solution of allyl glycoside 37 (700 mg, 770 mol, 1.0 equiv.) in anhyd. THF (10 mL). After stirring for 2 h at rt, NIS (191 mg, 847 mol, 1.1 equiv.) and H.sub.2O (12 mL) were added. After stirring for an additional hour, a TLC analysis (Tol/EtOAc 7:3) showed the complete consumption of disaccharide 37 (R.sub.f 0.45) and the presence of a polar spot (R.sub.f 0.2). 10% Aq. Na.sub.2SO.sub.3 was added. Volatiles were removed under reduced pressure and the aq. phase was extracted with DCM (20 mL) twice. The combined organic layers were washed with brine, dried over anhyd. Na.sub.2SO.sub.4, and concentrated. Purification of the residue by flash chromatography (cHex/EtOAc 5:1.fwdarw.4:1) gave the expected hemiacetal 38 (620 mg, 713 mol, 92%) as a white solid. Hemiacetal 38, isolated as a 10:7 cc/p mixture had R.sub.f 0.15 (Tol/EtOAc 1:1). The major anomer had .sup.1H NMR (CDCl.sub.3) 7.84-7.26 (m, 17H, H.sub.Ar), 6.94 (d, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.90-5.80 (m, 1H, CH.sub.All), 5.73 (d, J.sub.2,NH=6.8 Hz, NH.sub.A), 5.33 (d, 1H, J.sub.1,2=5.6 Hz, H-1.sub.A), 5.27-5.22 (m, 1H, CH.sub.2All), 5.20-5.14 (m.sub.po, 3H, CH.sub.2All, CH.sub.2Bn-6), 4.80 (d, 1H, J.sub.4,5=4.4 Hz, H-5.sub.A), 4.77 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.70 (brs, 2H, CH.sub.2Nap), 4.53-4.46 (m, 3H, H-3.sub.B, CH.sub.2Bn), 4.33-4.28 (m, 1H, CH.sub.2All), 4.10 (dd.sub.po, J.sub.3,4=2.8 Hz, H-4.sub.A), 4.07-4.00 (m, 3H, H-3.sub.A, H-2.sub.A, CH.sub.2All), 3.93 (brd, 1H, J.sub.3,4=3.2 Hz, H-4.sub.B), 3.57 (ddd.sub.po, 1H, J.sub.2,3=9.2 Hz, H-2.sub.B), 3.47 (brq, 1H, H-5.sub.B), 1.86 (s, 3H, CH.sub.3NAc), 1.23 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 170.4 (CO.sub.NTCA), 169.3 (C-6.sub.A), 161.9 (CO.sub.NAc), 137.9, 134.9, 134.8, 133.1, 133.0 (C.sub.q, Ar), 133.5 (CH.sub.All), 128.7 (2C), 128.6, 128.3, 128.1, 127.9, 127.8, 127.6, 126.9, 126.1, 126.0, 125.9 (C.sub.Ar), 117.9 (CH.sub.2All), 99.5 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.4 (CCl.sub.3), 76.6 (C-3.sub.B), 73.4 (C-3.sub.A), 72.9 (C-4.sub.A), 71.9 (C-5.sub.A), 71.8 (2C, CH.sub.2Bn, CH.sub.2Nap), 70.1 (CH.sub.2All), 69.3 (C-5.sub.B), 67.5 (CH.sub.2Bn-6), 65.1 (C-4.sub.B), 55.0 (C-2.sub.B), 52.1 (C-2.sub.A), 23.4 (CH.sub.3Ac), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.41H.sub.43Cl.sub.3N.sub.5O.sub.10, 870.2076; found 870.2070.

    [0535] (Benzyl 2-acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido-/-D-galactopyranosyl (N-phenyl)trifluoroacetimidate (39) and 2-Trichloromethyl-[(benzyl 2-acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-1,2,4,6-tetradeoxy--D-galactopyrano]-[2,1,d]-oxazoline (40). Hemiacetal 38 (630 mg, 725 mol, 1.0 equiv.) was dissolved in acetone (10 mL). PTFACl (149 l, 942 mol, 1.3 equiv.) was added followed by Cs.sub.2CO.sub.3 (260 mg, 797 mol, 1.1 equiv.). The reaction mixture was stirred for 2 h at rt under an Ar atmosphere. A TLC follow up (Tol/EtOAc 5:1) showed that the starting 38 had evolved into two less polar spots (R.sub.f 0.3 and 0.35). The reaction mixture was filtered over a pad of Celite, washed with acetone (10 mL) twice and the filtrate was concentrated under reduced pressure. The residue was purified by rapid flash chromatography (cHex/EtOAc 4:1.fwdarw.2:1, 1% Et.sub.3N) to give the desired donor as a 3:2 mix of 39 and 40 (670 mg, 644 mol, 89%) isolated as a floppy white solid. The isolated mix of donors 39 and 40 had .sup.1H NMR (CDCl.sub.3) 7.87-6.80 (m, 22H.sub.Ar,PTFA, NH.sub.B,PTFA, 17H-.sub.Ar,oxa), 6.56 (bs, 0.6H, H-1.sub.B,PTFA), 6.18 (d, 0.4H, J.sub.1,2=7.0 Hz, H-1.sub.B,oxa), 5.86 (d, 0.4H, J.sub.2,NH=7.2 Hz, NH.sub.A,oxa), 5.48 (d, 0.4H, J.sub.1,2=5.6 Hz, H-1.sub.A,oxa), 5.41 (d.sub.po, 0.6H, J.sub.1,2=7.6 Hz, H-1.sub.A,PTFA), 5.39 (d.sub.po, J.sub.1,2=8.0 Hz, NH.sub.A,PTFA), 5.23 (d, 0.4H, J=12 Hz, CH.sub.2Bn-6), 5.16 (s.sub.po, 1.2H, CH.sub.2Bn-6), 5.14 (d.sub.po, 0.4H, CH.sub.2Bn-6), 4.87 (d, 0.6H, J=12.5 Hz, CH.sub.2Nap), 4.81 (d, 0.6H, CH.sub.2Nap), 4.78 (d.sub.po, 0.6H, J.sub.3,4=3.0 Hz, H-5.sub.A), 4.76 (d.sub.po, 0.6H, J.sub.3,4=4.9 Hz, H-5.sub.A), 4.69 (d, 0.4H, J=12.4 Hz, CH.sub.2Nap), 4.64 (d, 0.4H, CH.sub.2Nap), 4.59 (d, 0.4H, J=12.1 Hz, CH.sub.2Bn), 4.54 (d, 0.4H, CH.sub.2Bn), 4.48 (dd, 0.6H, J.sub.3,4=3.1 Hz, J.sub.3,4=11.0 Hz, H-3.sub.B), 4.42 (do, 0.6H, J=12.0 Hz, CH.sub.2Bn), 4.42-4.37 (m.sub.o, 0.6H, H-2.sub.B), 4.30 (ddd, 0.6H, H-2.sub.A), 4.26 (bs, 0.6H, H-4.sub.B), 4.20-4.23 (m, 2.2H, CH.sub.2Bn, H-3.sub.A, H-4.sub.A, H-2.sub.B), 4.10 (dd.sub.po, 0.6H, H-4.sub.A), 4.08-4.03 (m.sub.po, 0.4H, H-2.sub.A), 4.05-4.99 (m, 0.6H, H-5.sub.B), 3.86 (m, 0.4H, H-4.sub.B), 3.75 (dd, 0.4H, H-5.sub.B), 3.57 (dd, 0.4H, J.sub.3,4=3.7 Hz, J.sub.2,3=8.1 Hz, H-3.sub.B), 3.54 (dd, 0.6H, J.sub.2,3=10.1 Hz, J.sub.3,4=2.8 Hz, H-3.sub.A), 1.94 (s, 1.8H, CH.sub.3Ac), 1.93 (s, 1.2H, CH.sub.3Ac), 1.28 (d.sub.po, 0.6H, J.sub.5,6=6.3 Hz, H-6.sub.B), 1.27 (d.sub.po, 0.4H, J.sub.5,6=6.3 Hz, H-6.sub.B). .sup.13C NMR (Partial, CDCl.sub.3) 173.2, 170.7, 170.5 (CO.sub.NTCA), 169.3, 168.9 (C-6.sub.A), 162.9, 162.1 (CO.sub.NAc), 143.0, 137.9, 137.3, 135.0, 134.9, 134.7, 134.6, 133.1, 133.0 (C.sub.q,Ar), 129.1, 128.8 (2C), 128.6, 128.5, 128.4, 128.3 (2C), 128.2, 128.0 (2C), 127.9 (2C), 127.8, 127.6, 127.2, 126.9, 126.8, 126.2, 126.1 (2C), 126.0 (2C), 125.9, 124.4, 119.4, 118.3 (C.sub.Ar), 107.6 (C-1.sub.B,oxa, .sup.1J.sub.C,H=183 Hz), 99.0 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 98.8 (C-1.sub.A, .sup.1J.sub.C,H=171 Hz), 93.7 (bs, C-1.sub.B-PTFA), 92.4 (CCl.sub.3), 86.9 (CCl.sub.3), 81.1 (C-3.sub.A), 74.5 (C-3.sub.B), 73.6 (C-5.sub.A), 73.1 (C-4.sub.A), 73.0 (C-4.sub.A), 72.5 (C-3.sub.B), 72.1 (CH.sub.2Nap), 71.8 (2C, CH.sub.2Nap, CH.sub.2Bn), 71.5 (C-5.sub.A), 71.4 (2C, C-3.sub.A, CH.sub.2Bn), 69.0 (C-5.sub.B), 68.0 (C-5.sub.B), 67.4 (2C, CH.sub.2Bn-6), 64.2 (C-4.sub.B), 63.5 (C-2.sub.B), 61.3 (C-4.sub.B), 52.3 (C-2.sub.A), 51.0 (C-2.sub.A), 50.3 (C-2.sub.B), 23.7, 23.4 (CH.sub.3Ac), 17.5, 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.49H.sub.47Cl.sub.3F.sub.3N.sub.6O.sub.10, 1041.2372; found 1041.2378.

    [0536] 3-Azidopropyl (benzyl 2-acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (31). Freshly activated MS 4 (50 mg) was added to donors 39/40 (2:1, 50 mg, 48 mol, 1.0 equiv.) in anhyd. DCM (3.0 mL) containing 3-azidopropanol (22 L, 240 mol, 5.0 equiv.). The reaction mixture was stirred for 45 min at rt in an Ar atmosphere and cooled to 0 C. Yb(OTf).sub.3 (3.0 mg, 5.0 mol, 0.1 equiv.) was added and after stirring for 30 min at 0 C., a TLC analysis (Tol/EtOAc 7:3) confirmed the absence of donors 39/40 (R.sub.f 0.45, 0.55) and the presence of a major compound (R.sub.f 0.2). Et.sub.3N (2.0 L) was added and the suspension was filtered over a fitted funnel, and washed thoroughly with DCM. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography (Tol/EtOAc 60:40.fwdarw.50:50) to give the condensation product 41 (36 mg, 37 mol, 78%) as a white solid. The azidopropyl glycoside 41 had R.sub.f 0.2 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.85-7.71 (m, 4H, H.sub.Ar), 7.51-7.47 (m, 2H, H.sub.Ar), 7.42-7.25 (m, 11H, H.sub.Ar), 6.83 (d, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.55 (d, J.sub.2,NH=7.2 Hz, NH.sub.A), 5.28 (d, 1H, J.sub.1,2=5.9 Hz, H-1.sub.A), 5.22 (d, H, J=12.0 Hz, CH.sub.2Bn-6), 5.19 (d, H, CH.sub.2Bn-6), 4.82 (d, 1H, J.sub.4,5=4.8 Hz, H-5.sub.A), 4.75 (d, H, J=12.5 Hz, CH.sub.2Nap), 5.22 (d, H, CH.sub.2Nap), 4.65 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.53 (d, 1H, J=11.9 Hz, CH.sub.2Bn), 4.45 (d, 1H, CH.sub.2Bn), 4.39 (dd, 1H, J.sub.3,4=3.5 Hz, J.sub.2,3=10.8 Hz, H-3.sub.B), 4.11 (dd, J.sub.3,4=2.8 Hz, H-4.sub.A), 4.07 (pdt, 1H, H-2.sub.A), 3.97 (dd.sub.po, 1H, J.sub.2,3=8.2 Hz, H-3.sub.A), 3.93-3.88 (m, 2H, H-4.sub.B, OCH.sub.2), 3.62 (ddd, 1H, H-2.sub.B), 3.56-3.51 (m, 1H, OCH.sub.2), 3.49 (brq, 1H, H-5.sub.B), 3.38 (t, 2H, J=6.8 Hz, NCH.sub.2), 1.89-1.78 (m, 2H, CH.sub.2), 1.87 (s, 3H, CH.sub.3NHAc), 1.23 (d, 3H, J.sub.5,6=6.2 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 170.3 (CO.sub.NHTCA), 169.3 (C-6.sub.A), 161.8 (CO.sub.NHAc), 137.8, 134.9, 134.8, 133.1, 133.0 (C.sub.q,Ar), 128.7, 128.6, 128.3, 128.2, 128.1, 127.8 (2C), 127.7, 126.8, 126.1, 126.0, 125.9 (C.sub.Ar), 99.5 (C-1.sub.A, .sup.1J.sub.C,H=171 Hz), 97.2 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.5 (CCl.sub.3), 76.5 (C-3.sub.B), 73.6 (C-3.sub.A), 72.7 (C-4.sub.A), 71.9 (C-5.sub.A), 71.8 (2C, CH.sub.2Bn, CH.sub.2Nap), 69.4 (C-5.sub.B), 67.4 (CH.sub.2Bn-6), 66.3 (OCH.sub.2), 64.9 (C-4.sub.B), 54.7 (C-2.sub.B), 52.0 (C-2.sub.A), 48.1 (CH.sub.2N.sub.3), 29.0 (CH.sub.2), 23.5 (CH.sub.3Ac), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.44H.sub.48Cl.sub.3N.sub.8O.sub.10, 953.2559; found 953.2542.

    Example 2: Strategy 2.SUB.A.-NAcBoc,2.SUB.B.-NTCA, 4.SUB.A.-Nap Series

    [0537] Use of an Acid-Sensitive Acetamide Camouflage of the 2.sub.A-NHAc

    ##STR00090##

    ##STR00091##

    [0538] Allyl (benzyl 2-(N-tert-butyloxycarbonyl)acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (57). Di-tert-butylcarbonate (1.22 g, 5.6 mmol, 8.0 equiv.) followed by DMAP (34 mg, 282 mol, 0.4 equiv.) were added to disaccharide 37 (640 mg, 704 mol, 1.0 equiv.) in anhyd. THF (20 mL). After heating at 50 C. for 2 h, a TLC follow up (Tol/EtOAc 4:1) showed the presence of a less polar spot (R.sub.f 0.75) and the absence of the starting 37 (R.sub.f 0.1). The reaction mixture was allowed to reach rt and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 10:1.fwdarw.7:1) to give the desired 57 as a white solid (520 mg, 515 mol, 73%). Disaccharide 57 had .sup.1H NMR (CDCl.sub.3) 7.84-7.74 (m, 4H, H.sub.Ar), 7.49-7.46 (m, 3H, H.sub.Ar), 7.39-7.36 (m, 5H, H.sub.Ar), 7.29-7.18 (m, 5H, H.sub.Ar), 6.75 (d, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.92-5.82 (m, 1H, CH.sub.All), 5.72 (d, 1H, J.sub.1,2=8.2 Hz, H-1.sub.A), 5.29-5.23 (m, 1H, CH.sub.2All), 5.22 (d.sub.po, 1H, CH.sub.2Bn-6), 5.19 (d.sub.po, 1H, J=12.1 Hz, CH.sub.2Bn-6), 5.20-5.16 (m.sub.po, 1H, CH.sub.2All), 5.00-4.89 (br, 1H, H-2.sub.A), 4.85 (d.sub.po, 1H, J=12.3 Hz, CH.sub.2Bn), 4.81 (d.sub.po, 1H, CH.sub.2Bn), 4.75 (d.sub.po, 1H, J.sub.1,2=8.3 Hz, H-1.sub.B), 4.72 (d, J.sub.4,5=2.6 Hz, H-5.sub.A), 4.42 (d.sub.po, 1H, J=11.8 Hz, CH.sub.2Nap), 4.38 (dd.sub.po, 2H, J.sub.2,3=10.5 Hz, J.sub.3,4=2.3 Hz, H-3.sub.B), 4.37 (pt.sub.o, 1H, J.sub.3,4=2.8 Hz, H-4.sub.A), 4.34-4.29 (m, 2H, CH.sub.2Nap, CH.sub.2All), 4.24 (brd, 1H, J.sub.2,3=10.4 Hz, H-3.sub.A), 4.05 (bd, 1H, H-4.sub.B), 4.05-4.00 (m, 1H, CH.sub.2All), 3.61-3.54 (pdt, 1H, H-2.sub.B), 3.44 (dq, 1H, J.sub.4,5=1.0 Hz, H-5.sub.B), 2.34 (s, 3H, CH.sub.3NAc), 1.48 (s, 9H, CH.sub.3NBoc), 1.26 (d, 3H, J.sub.5,6=6.2 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 174.4 (br, CO.sub.NAc), 169.1 (C-6.sub.A), 161.7 (CO.sub.NTCA), 153.9 (br, CO.sub.NBoc), 137.8, 135.5, 135.0, 133.2, 132.9 (C.sub.q,Ar), 133.6 (CH.sub.All), 129.0, 128.8, 128.7 (2C), 128.2 (2C), 127.8, 127.6, 126.2, 126.0, 125.8, 125.7, 125.2 (10C, C.sub.Ar), 117.9 (CH.sub.2All), 98.9 (C-1.sub.A, .sup.1J.sub.C,H=174 Hz), 97.9 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 92.4 (CCl.sub.3), 83.7 (C.sub.Boc), 76.9 (br, C-3.sub.B), 74.1 (2C, C-4.sub.A, C-5.sub.A), 73.7 (br, C-3.sub.A), 72.5 (CH.sub.2Nap), 71.7 (CH.sub.2Bn), 70.1 (CH.sub.2All), 68.8 (C-5.sub.B), 67.4 (CH.sub.2Bn-6), 65.3 (C-4.sub.B), 55.0 (C-2.sub.B), 27.8 (CH.sub.3Boc), 27.1 (CH.sub.3Ac), 17.4 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.49H.sub.58Cl.sub.3N.sub.6O.sub.12, 1027.3179; found 1027.3176.

    [0539] Allyl (benzyl 2-(N-tert-butyloxycarbonyl)acetamido-3-O-benzyl-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (58). DDQ (256 mg, 1.1 mmol, 3.0 equiv.) was added to the fully protected disaccharide 57 (380 mg, 377 mol, 1.0 equiv.) in DCM/phosphate buffer pH 7 (10:1, 10 mL). After stirring vigorously at rt for 2 h, TLC analysis (Tol/EtOAc 4:1) revealed that the starting 57 (R.sub.f 0.75) had evolved into a more polar product (R.sub.f 0.35). 5% Aq. NaHCO.sub.3 (10 mL) and DCM (10 mL) were added. The DCM layer was separated, washed with brine (25 mL), dried over Na.sub.2SO.sub.4, and concentrated. The residue was purified by flash chromatography (Tol/EtOAc 5:1.fwdarw.4:1) to give alcohol 58 as a white solid (260 mg, 257 mol, 79%). Disaccharide 58 had .sup.1H NMR (CDCl.sub.3) 7.44-7.40 (m, 5H, H.sub.Ar), 7.32-7.22 (m, 5H, H.sub.Ar), 6.65 (d, J.sub.2,NH=7.6 Hz, NH.sub.B), 5.91-5.81 (m, 1H, CH.sub.All), 5.61 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A), 5.28-5.22 (m.sub.po, 1H, CH.sub.2All), 5.25 (s, 2H, CH.sub.2Bn-6), 5.19-5.15 (m, 1H, CH.sub.2All), 4.71 (d.sub.po, 1H, J.sub.4,5=2.6 Hz, H-5.sub.A), 4.70-4.62 (br, 1H, H-2.sub.A), 4.65 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.55-4.53 (m.sub.po, 1H, H-4.sub.A), 4.54 (d.sub.po, 1H, CH.sub.2Bn), 4.42 (d, 1H, J=11.6 Hz, CH.sub.2Bn), 4.35 (dd, 1H, J.sub.2,3=10.8 Hz, J.sub.3,4=3.7 Hz, H-3.sub.B), 4.33-4.27 (m, 2H, H-3.sub.A, CH.sub.2All), 4.04-3.99 (m, 2H, H-4.sub.B, CH.sub.2All), 3.61 (pt, 1H, H-2.sub.B), 3.39 (dq, 1H, J.sub.4,5=1.0 Hz, H-5.sub.B), 2.59 (d, 1H, J.sub.4,OH=2.2 Hz, OH), 2.31 (s, 3H, CH.sub.3Ac), 1.50 (s, 9H, CH.sub.3Boc), 1.25 (d, 3H, J.sub.5,6=6.3 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 174.1 (br, CO.sub.NAc), 168.8 (C-6.sub.A), 161.7 (CO.sub.NTCA), 153.6 (br, CO.sub.NBoc), 137.2, 135.0 (C.sub.q,Ar), 133.6 (CH.sub.All), 128.8 (2C), 128.5, 128.1, 127.8, (10C, C.sub.Ar), 117.9 (CH.sub.2All), 98.5 (C-1.sub.A, .sup.1J.sub.C,H=174 Hz), 98.1 (C-1.sub.B, .sup.1J.sub.C,H=161 Hz), 92.4 (CCl.sub.3), 83.9 (OC.sub.Boc), 76.8 (C-3.sub.B), 75.2 (C-5.sub.A), 73.3 (br, C-3.sub.A), 72.0 (CH.sub.2Bn), 70.1 (CH.sub.2All), 68.7 (C-5.sub.B), 67.6 (CH.sub.2Bn-6), 66.8 (C-4.sub.A), 65.3 (C-4.sub.B), 54.9 (C-2.sub.B), 27.8 (CH.sub.3Boc), 27.1 (CH.sub.3Ac), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.38H.sub.50Cl.sub.3N.sub.6O.sub.12, 887.2552; found 887.2558.

    [0540] (Benzyl 2-(N-tert-butyloxycarbonyl)acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido-/-D-galactopyranose (59). A solution of [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (12 mg, 13 mol, 0.03 equiv.) in anhyd. THF (3.0 mL), was degassed and stirred for 20 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly with Ar and poured into a solution of allyl glycoside 57 (430 mg, 473 mol, 1.0 equiv.) in anhyd. THF (16 mL). After stirring for 1 h at rt, NIS (117 mg, 520 mol, 1.1 equiv.) and H.sub.2O (4.0 mL) were added. After stirring for another hour at rt, a TLC follow up (CHex/EtOAc 6:1) showed the absence of allyl glycoside 57 (R.sub.f 0.6) and products in close vicinity. 10% Aq. Na.sub.2SO.sub.3 was added, and volatiles were evaporated. The aq. phase was extracted with DCM (10 mL) twice. The combined organic layers were washed with brine (30 mL), dried over anhyd. Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. Purification of the residue by flash chromatography (Chex/EtOAc 70:30.fwdarw.65:35) gave the expected hemiacetal 59 (360 mg, 356 mol, 87%) as a white solid. Hemiacetal 59 had R.sub.f 0.15, 0.25 (Tol/EtOAc, 3:1). .sup.1H NMR (major isomer, CDCl.sub.3) 7.84-7.17 (m, 17H, H.sub.Ar), 6.65 (d, 1H, J.sub.2,NH=7.6 Hz, NH.sub.B), 5.85 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A), 5.40 (brs, 1H, H-1.sub.B), 5.24 (d, 1H, CH.sub.2Bn-6), 5.16 (d, 1H, J=12.0 Hz, CH.sub.2Bn-6), 5.05 (brs, 1H, H-2.sub.A), 4.90 (d, 1H, CH.sub.2Nap), 4.82 (d, 1H, J=12.3 Hz, CH.sub.2Nap), 4.72 (d, 1H, J.sub.4,5=2.4 Hz, H-5.sub.A), 4.41 (d, 1H, J=11.6 Hz, CH.sub.2Bn), 4.36 (d.sub.po, 1H, H-4.sub.A), 4.35-4.26 (m, 2H, H-3.sub.B, H-2.sub.B), 4.20 (dd, 1H, J.sub.2,3=10.5 Hz, J.sub.3,4=2.1 Hz, H-3.sub.A), 4.13 (brs.sub.po, 1H, H-4.sub.B), 4.11 (bq.sub.po, 1H, H-5.sub.B), 3.15 (brs, 1H, OH), 2.36 (s, 3H, CH.sub.3NAc), 1.47 (s.sub.po, 9H, CH.sub.3Boc), 1.22 (d, 3H, J.sub.5,6=6.2 Hz, H-6.sub.B). .sup.13C NMR (major isomer, CDCl.sub.3) 174.7 (br, CO.sub.NAcBoc), 168.9 (C-6.sub.A), 162.0 (CO.sub.NTCA), 153.4 (br, CO.sub.Boc), 137.7, 137.5, 135.4, 135.1, 134.8, 133.2, 133.0 (C.sub.q,Ar), 128.9, 128.8, 128.7 (2C), 128.5, 128.3 (2C), 128.1, 127.8, 127.7, 126.3, 126.2, 126.0, 125.9, 125.7 (C.sub.Ar), 98.0 (br, C-1.sub.A, .sup.1J.sub.C,H=174 Hz), 92.4 (CCl.sub.3), 90.8 (C-1.sub.B, .sup.1J.sub.C,H=174 Hz), 84.1 (C.sub.Boc), 74.2 (C-5.sub.A), 74.0, 73.9 (2br, 3C, C-3.sub.B, C-4.sub.A, C-3.sub.A), 72.7 (CH.sub.2Nap), 71.6 (CH.sub.2Bn), 67.4 (CH.sub.2Bn-6), 65.6 (C-4.sub.B), 64.6 (C-5.sub.B), 55.4 (br, C-2.sub.A), 51.1 (C-2.sub.B), 27.8 (CH.sub.3NBoc), 27.4 (CH.sub.3Ac), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.46H.sub.58Cl.sub.3N.sub.7O.sub.12, 987.2860; found 987.2862.

    [0541] (Benzyl 2-(N-tert-butyloxycarbonyl)acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido-/-D-galactopyranosyl)N-(phenyl)trifluoroacetamidate (60). PTFACl (23 L, 148 mol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (197 mg, 604 mol, 1.1 equiv.) were added to hemiacetal 59 (110 mg, 113 mol, 1.0 equiv.) in acetone (4.0 mL). After stirring at rt for 2 h under an Ar atmosphere, a TLC follow up (Tol/EtOAc 6:1) indicated that hemiacetal 59 (R.sub.f 0.1) had been converted to a less polar compound (R.sub.f 0.85). The suspension was filtered over a pad of Celite, solids were washed with DCM (4 mL) twice, and volatiles were evaporated. The crude residue was purified by flash chromatography (Chex/EtOAc 90:10.fwdarw.98:12, 1% Et.sub.3N) to give donor 60 (110 mg, 96 mol, 86%) as a white solid. The donor had .sup.1H NMR (main isomer, CDCl.sub.3) 7.85-7.10 (m, 21H, H.sub.Ar), 6.79 (d, 2.2H, J=7.6 Hz), 6.53 (brs, 0.9H), 5.94 (d, 1H, J=8.4 Hz), 5.24 (d, 1H, CH.sub.2Bn-6), 5.16 (d, 1H, J=12.0 Hz, CH.sub.2Bn-6), 4.90 (brs, 2H, CH.sub.2Nap), 4.74 (d, 1H, J=2.0 Hz, H-5.sub.A), 4.37 (d.sub.po, 2H, H-2.sub.A, CH.sub.2Bn), 4.36-4.33 (m, 3H, H-3.sub.A, H-4.sub.A, H-4.sub.B), 4.23 (d.sub.po, 1H, J=11.9 Hz, CH.sub.2Bn), 4.22-4.17 (m, 2H, H-2.sub.B, H-3.sub.B), 3.97-3.91 (dq, 1H, H-5.sub.B), 2.37 (s, 3H, CH.sub.3Ac), 1.47 (s, 9H, CH.sub.3Boc), 1.25 (d, 3H, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.6, 162.1, 143.0, 138.0, 137.5, 135.0, 134.7, 133.2, 133.0 (C.sub.q,Ar), 129.4, 129.0, 128.8, 128.7, 128.6 (2C), 128.5, 128.3, 128.2 (2C), 128.0, 127.9, 127.8 (2C), 127.6, 126.8, 126.3, 126.1, 126.0, 125.9, 125.7, 125.2, 124.4, 120.4, 119.3 (C.sub.Ar), 107.3, 97.0, 93.4, 92.1, 84.4 (C.sub.Boc), 77.2, 74.5 (C-5.sub.A), 74.2 (2C), 72.5 (CH.sub.2Nap), 71.6 (CH.sub.2Bn), 67.7 (C-5.sub.B), 67.5 (CH.sub.2Bn-6), 64.5 (C-2.sub.A), 50.3 (C-2.sub.B), 27.9, 27.8, 27.3, 26.9, 17.6, 17.3. HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.54H.sub.58C.sub.13F.sub.3N.sub.7O.sub.12, 1158.3156; found 1158.3137.

    [0542] Allyl (benzyl 2-(N-tert-butyloxycarbonyl)acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranyl)-(1.fwdarw.4)-(benzyl 2-(N-tert-butyloxycarbonyl)acetamido-3-O-benzyl-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (61). A mix of PTFA donor 60 (58 mg, 51 mol, 1.1 equiv.) and acceptor 58 (40 mg, 46 mol, 1.0 equiv.) was coevaporated with anhyd. toluene, dried under vacuum thoroughly, and taken into anhyd. DCE (2.0 mL) containing activated MS 4 (100 mg). The reaction mixture was stirred at rt for 30 min under an Ar atmosphere and cooled to 0 C. TfOH (0.2 L, 0.05 equiv.) in 10 L ACN was added. After stirring at this temperature for another 30 min, a TLC analysis (Tol/EtOAc 4:1) showed the absence of donor 60 and the presence of a new spot. Et.sub.3N was added and solids were filtered off. Volatiles were evaporated and the crude was purified by flash chromatography (Tol/EtOAc 85:15.fwdarw.80:20) to give the desired 61 (35 mg, 19 mol, 41%) as a white solid. Tetrasaccharide 61 had R.sub.f 0.35 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.83-7.75 (m, 5H, H.sub.Ar), 7.48-7.17 (m, 24H, H.sub.Ar), 6.85 (brs, 1H, NH.sub.B*), 6.85 (brs, 1H, NH.sub.B1*), 5.89-5.83 (m, 1H, CH.sub.All), 5.73 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A*), 5.53 (brd, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A1*), 5.28-5.15 (m, 6H, 2CH.sub.2Bn-6, CH.sub.2All), 5.03-4.88 (brs, 2H, H-2.sub.A*, H-1.sub.B1), 4.82 (brs, 3H, H-5.sub.A*, CH.sub.2Nap), 4.71-4.68 (m, 2H, H-5.sub.A1*, H-1.sub.B), 4.56-4.53 (m, 2H, H-2.sub.A1*, H-3.sub.B*), 4.44-4.21 (m, 9H, H-3.sub.B1*, H-3.sub.A*, H-3.sub.A1*, H-4.sub.A*, H-4.sub.A1*, 2CH.sub.2Bn), 4.07-4.00 (m, 3H, H-4.sub.B, CH.sub.2All), 3.64-3.38 (m, 1H, H-2.sub.B, H-5.sub.B), 2.38-2.22 (s, 6H, CH.sub.3Ac), 1.52-1.43 (m, 18H, CH.sub.3Boc), 1.29 (brd, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B*), 1.19 (brd, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B1*). .sup.13C NMR (CDCl.sub.3) 174.4 (CO.sub.NAcBoc), 169.2, 168.7 (2C, C-6.sub.A, C-6.sub.A1), 161.9, 161.6 (2C, CO.sub.NTCA), 153.8 (2C, CO.sub.NAcBoc), 137.9, 137.8, 135.5, 135.1, 15.0, 133.2, 132.9 (C.sub.q, Ar), 133.7 (CH.sub.All), 129.0, 128.9, 128.8, 128.7, 128.6 (2C), 128.3, 128.2 (2C), 127.9, 127.8, 127.7, 127.6 (2C), 126.2, 126.0, 125.8, 125.7, 125.2 (10C, C.sub.Ar), 117.6 (CH.sub.2All), 99.4 (C-1.sub.B*, .sup.1J.sub.C,H=169 Hz), 98.9 (C-1.sub.A*, .sup.1J.sub.C,H=175 Hz), 98.5 (2C, C-1.sub.B1*, .sup.1J.sub.C,H=164 Hz, C-1.sub.A1*, .sup.1J.sub.C,H=177 Hz), 92.4 (CCl.sub.3), 92.1 (CCl.sub.3), 83.8 (C.sub.Boc), 83.6 (C.sub.Boc), 77.6, 77.2 (2C, C-3.sub.B, C-3.sub.B1), 76.0, 74.1 (2C, C-5.sub.A, C-5.sub.A1), 73.6 (2C, C-3.sub.A, C-3.sub.A1), 72.5 (CH.sub.2Nap), 71.7, 71.6 (2C, CH.sub.2Bn), 70.0 (CH.sub.2All), 68.8 (2C, C-5.sub.B, C-5.sub.B1), 67.4, 67.3 (2C, CH.sub.2Bn-6), 66.8 (2C, C-4.sub.A, C-4.sub.A1), 65.2 (2C, C-4.sub.B, C-4.sub.B1), 55.4 (2C, C-2.sub.A, C-2.sub.A1), 54.7 (2C, C-2.sub.B, C-2.sub.B1), 27.9, 27.8 (2C, CH.sub.3NBoc), 27.4, 27.2 (2C, CH.sub.3Ac), 17.4, 17.2 (2C, C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.84H.sub.98Cl.sub.6N.sub.11O.sub.23 1838.4963; found 1838.4982.

    [0543] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (1)..sup.[1] Route 1. TFA (15 L, 197 mol, 4 equiv.) was added to a solution of disaccharide 57 (46 mg, 49 mol, 1.0 equiv.) in DCM (2 mL) at rt. After 3-4 h, a TLC follow up with Tol/EtOAc (4:1) showed the presence of a more polar single spot. Dilute aq. NaHCO.sub.3 (5 mL) was added followed by DCM (5 mL). The DCM layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. 20% Pd(OH).sub.2/C (68 mg) was added to a solution of the crude intermediate in .sup.tBuOH/DCM/H.sub.2O (16 mL, 7:3:1) and after extensive degassing, the atmosphere was saturated with hydrogen. After stirring under hydrogen for 48 h, the suspension was passed through a 0.2 m filter and washed thoroughly with methanol. Volatiles were evaporated and the crude intermediate was dissolved in water (5 mL) and lyophilized. Purification by semi-preparative RP-HPLC gave the known propyl glycoside 1 as a white solid (16 mg, 35 mol, 71%).

    [0544] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (2). Tetrasaccharide 61 (15 mg, 8.2 mol, 1.0 equiv.), contaminated to a 15-20% extent by the disaccharide partners, was dissolved in DCM (1.0 mL) and TFA (15 L, 197 mol, 24 equiv.) was added. After stirring for 3 h at rt, a TLC follow up (Tol/EtOAc 2:1) indicated reaction completion. 10% Aq. NaHCO.sub.3 (5 mL) and DCM (5 mL) were added. The organic layer was separated, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The crude was dissolved in .sup.tBuOH/DCM/H.sub.2O (11 mL, 7:3:1) and 20% Pd(OH).sub.2/C (50 mg) was added. After stirring under an atmosphere of hydrogen for 48 h, the suspension was passed through a syringe filter (0.2 m) and washed thoroughly with methanol. The filtrate was evaporated and the crude was dissolved in water (5 mL) and lyophilized. The residue was purified by semi-preparative RP-HPLC to give the propyl glycoside 2 as a white foam (2.6 mg, 3.0 mol, 37% (underestimated)). Tetrasaccharide 2 had RP-HPLC (215 nm) R.sub.t=12.3 min (conditions A), R.sub.t=13.6 min (conditions B), .sup.1H NMR (D.sub.2O) 4.87 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A), 4.77 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A1), 4.74 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A1), 4.66 (brs, 1H, H-5.sub.A), 4.44 (d.sub.po, 2H, J=8.4 Hz, H-1.sub.B1, H-4.sub.A), 4.36 (brs, 1H, H-4.sub.A1), 4.15-4.08 (m, 2H, H-3.sub.B, H-3.sub.B1), 4.03-3.99 (m.sub.po, 2H, H-5.sub.B, H-5.sub.B1), 3.96-3.90 (m, 2H, H-2.sub.A, H-4.sub.B), 3.80-3.66 (m, 7H, H-3.sub.A, H-3.sub.A1, H-2.sub.A1, H-2.sub.B, H-2.sub.B1, H-4.sub.B1, OCH.sub.2Pr), 3.51-3.49 (m.sub.po, 1H, OCH.sub.2Pr), 1.99, 1.94 (2s, 12H, CH.sub.3Ac), 1.51-1.48 (m, 2H, CH.sub.2Pr), 1.29 (d.sub.po, 6H, H-6.sub.B), 0.82 (t, 3H, J=7.2 Hz, CH.sub.3Pr). .sup.13C NMR (D.sub.2O) 174.6, 174.5 (2C), 174.0 (CO.sub.NHAc), 102.9 (C-1.sub.B1, .sup.1J.sub.C,H=168 Hz), 101.6 (C-1.sub.B, .sup.1J.sub.C,H=166 Hz), 101.1 (2C, C-1.sub.A, C-1.sub.A1, .sup.1J.sub.C,H=170 Hz, .sup.1J.sub.C,H=168 Hz), 79.7 (C-4.sub.A), 76.5, 76.3 (C-5.sub.A, C-5.sub.A1), 76.0 (2C, C-3.sub.B, C-3.sub.B1), 72.7 (OCH.sub.2Pr), 68.5 (C-4.sub.A1), 68.5 (C-3.sub.A1), 67.8 (C-3.sub.A), 67.6 (C-4.sub.A), 67.4, 67.3 (C-5.sub.B, C-5.sub.B1), 54.8 (C-4.sub.B), 54.7 (C-4.sub.B1), 51.5 (C-2.sub.A1), 51.3 (C-2.sub.A), 51.0, 50.8 (C-2.sub.B, C-2.sub.B1), 23.3, 22.2 (4C, CH.sub.3Ac), 21.2 (CH.sub.2Pr), 15.6, 15.5 (C-6.sub.B, C-6.sub.B1), 9.5 (CH.sub.3Pr). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.35H.sub.58N.sub.6O.sub.19Na, 889.3649; found 889.3636.

    Example 3: Strategy 2.SUB.A.-NAc.SUB.2.,2.SUB.B.-NTCA, 4.SUB.A.-Nap Series

    [0545] A Protecting Group Sensitive to Mild Base as Camouflage of the 2.sub.A-Acetamide

    ##STR00092##

    ##STR00093##

    [0546] Allyl (benzyl 3-O-benzyl-2-deoxy-2-(N,N-diacetyl)amino-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (47). DIPEA (9.5 mL, 54.9 mmol, 20.0 equiv.) and acetyl chloride (3.9 mL, 21.9 mmol, 20 equiv.) were added successively to a solution of disaccharide 37 (2.5 g, 2.75 mmol, 1.0 equiv.) in anhyd. DCM (90 mL) at 0 C. The mixture was allowed to reach rt slowly and was stirred overnight. A TLC follow up (Tol/EtOAc 4:1) showed the complete conversion of acetamido 37 (R.sub.f 0.25) to a less polar product (R.sub.f 0.8). 10% Aq. NaHCO.sub.3 (50 mL) was added and the biphasic mixture was diluted with DCM (20 mL). The organic layer was separated, dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 90:20.fwdarw.88:12) to give the fully protected 47 (2.36 g, 2.48 mmol, 90%) as an off-white solid. The N-acetylacetamido derivative 47 had R.sub.f 0.65 (Tol/EtOAc 7:3). .sup.1H NMR (CDCl.sub.3) 7.85-7.83 (m, 3H, H.sub.Ar), 7.52-7.45 (m, 3H, H.sub.Ar), 7.42-7.36 (m, 4H, H.sub.Ar), 7.30-7.26 (m, 4H, H.sub.Ar), 7.21-7.12 (m, 3H, H.sub.Ar), 6.75 (d, J.sub.2,NH=7.6 Hz, NH.sub.B), 5.92-5.82 (m, 1H, CH.sub.All), 5.80 (d, 1H, J.sub.1,2=7.8 Hz, H-1.sub.A), 5.29-5.23 (m, 1H, CH.sub.2All), 5.26 (d, H, J=11.9 Hz, CH.sub.2Bn-6), 5.21 (d, H, CH.sub.2Bn-6), 5.20-5.17 (m, 1H, CH.sub.2All), 4.85 (d, 1H, J=12.9 Hz, CH.sub.2Nap), 4.81 (d, 1H, CH.sub.2Nap), 4.76 (d, 1H, J.sub.1,2=8.3 Hz, H-1.sub.B), 4.73 (d, 1H, J.sub.4,5=2.2 Hz, H-5.sub.A), 4.48 (dd, 1H, J.sub.2,3=10.8 Hz, J.sub.3,4=3.8 Hz, H-3.sub.B), 4.39-4.34 (m.sub.po, 3H, H-4.sub.A, CH.sub.2Bn), 4.35-4.30 (m.sub.po, 1H, CH.sub.2All), 4.27 (dd.sub.po, 1H, J.sub.2,3=10.5 Hz, J.sub.3,4=2.9 Hz, H-3.sub.A), 4.26 (d.sub.po, 1H, J=11.6 Hz, CH.sub.2Bn), 4.06-4.00 (m.sub.po, 1H, CH.sub.2All), 4.05 (bd.sub.o, 1H, H-4.sub.B), 3.59 (ddd, 1H, H-2.sub.B), 3.45 (dq, 1H, J.sub.5,6=1.0 Hz, H-5.sub.B), 2.38 (s, 6H, CH.sub.3NAc), 1.27 (d, 3H, J.sub.5,6=6.3 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 175.1 (2C, CO.sub.NAc), 168.7 (C-6.sub.A), 161.9 (CO.sub.NTCA), 137.8, 137.4, 135.2, 134.9, 133.2, 133.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 128.7-125.2 (C.sub.Ar), 117.9 (CH.sub.2All), 98.8 (C-1.sub.A, .sup.1J.sub.C,H=176 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.3 (CCl.sub.3), 77.4 (C-3.sub.B), 73.9 (C-5.sub.A), 73.7 (C-3.sub.A), 73.0 (C-4.sub.A), 72.5 (CH.sub.2Nap), 71.9 (CH.sub.2Bn), 70.2 (CH.sub.2All), 68.7 (C-5.sub.B), 62.6 (CH.sub.2Bn-6), 65.2 (C-4.sub.B), 59.1 (C-2.sub.A), 55.2 (C-2.sub.B), 21.4 (CH.sub.3Ac), 17.4 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.46H.sub.52Cl.sub.3N.sub.6O.sub.11, 969.2759; found 969.2751.

    [0547] Allyl (benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido--D-galactopyranoside (48). Disaccharide 47 (1.0 g, 1.0 mmol, 1.0 equiv.) was dissolved in DCM (10 mL) and phosphate buffer pH 7 (1.0 mL) was added. The biphasic mixture was cooled to 0 C. and DDQ (477 mg, 2.1 mmol, 2.0 equiv.) was added. The reaction was slowly allowed to reach rt and stirred for 3 h at this temperature. At completion, 5% aq. NaHCO.sub.3 (30 mL) was added and the biphasic mixture was diluted with DCM (50 mL). The DCM layer was separated, washed with brine (100 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 5:1.fwdarw.4:1) to give alcohol 48 (800 mg, 0.93 mmol, 93%) as a white solid. Disaccharide 48 had R.sub.f 0.45 (Tol/EtOAc, 7:3). .sup.1H NMR (CDCl.sub.3) 7.45-7.39 (m, 5H, H.sub.Ar), 7.35-7.25 (m, 3H, H.sub.Ar), 7.20-7.17 (m, 2H, H.sub.Ar), 6.69 (d, J.sub.2,NH=7.4 Hz, NH.sub.B), 5.90-5.80 (m, 1H, CH.sub.All), 5.68 (d, 1H, J.sub.1,2=7.9 Hz, H-1.sub.A), 5.27 (s, 2H, CH.sub.2Bn-6), 5.28-5.22 (m, 1H, CH.sub.2All), 5.19-5.15 (m, 1H, CH.sub.2All), 4.73 (d, 1H, J.sub.4,5=2.3 Hz, H-5.sub.A), 4.67 (d, 1H, J.sub.1,2=8.3 Hz, H-1.sub.B), 4.53-4.52 (m.sub.po, 1H, H-4.sub.A), 4.50 (d.sub.po, 1H, CH.sub.2Bn), 4.45 (dd.sub.po, 1H, J.sub.2,3=10.7 Hz, J.sub.3,4=3.8 Hz, H-3.sub.B), 4.24 (d.sub.po, J=11.6 Hz, CH.sub.2Bn), 4.36 (dd, 1H, J.sub.2,3=10.3 Hz, J.sub.3,4=3.4 Hz, H-3.sub.A), 4.33-4.28 (m, 1H, CH.sub.All), 4.07-3.99 (m, 3H, H-2.sub.A, H-4.sub.B, CH.sub.2All), 3.60 (pdt, 1H, H-2.sub.B), 3.39 (dq.sub.po, 1H, J.sub.4,5=1.1 Hz, H-5.sub.B), 2.57 (d, 1H, J.sub.4,OH=2.0 Hz, OH), 2.37, 2.34 (2s, 6H, CH.sub.3NAc), 1.24 (d, 3H, J.sub.5,6=6.3 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 174.9 (2C, CO.sub.NAe), 168.3 (C-6.sub.A), 161.9 (CO.sub.NTCA), 136.8, 134.9 (C.sub.q,Ar), 133.5 (CH.sub.All), 129.0, 128.9, 128.8, 128.6, 128.3, 128.2, 128.0 (C.sub.Ar), 118.2 (CH.sub.2All), 98.3 (C-1.sub.A, .sup.1J.sub.C,H=174 Hz), 97.7 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.3 (CCl.sub.3), 72.2 (C-3.sub.B), 75.2 (C-5.sub.A), 72.7 (C-3.sub.A), 72.3 (CH.sub.2Bn), 70.1 (CH.sub.2All), 68.7 (C-5.sub.B), 67.8 (CH.sub.2Bn-6), 66.6 (C-4.sub.A), 65.2 (C-4.sub.B), 58.9 (C-2.sub.A), 55.0 (C-2.sub.B), 21.4 (CH.sub.3Ac), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.35H.sub.44Cl.sub.3N.sub.6O.sub.11, 829.2134; found 829.2128.

    [0548] (Benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-4-O-(2-naphthylmethyl)-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido-/-D-galactopyranose (49). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (18 mg, 0.02 mmol, 0.02 equiv.) in anhyd. THF (3.0 mL) was degassed and stirred for 20 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly with Ar and poured into a solution of allyl glycoside 47 (1.0 g, 1.05 mmol, 1.0 equiv.) in anhyd. THF (20 mL). After stirring for 1 h at rt, a TCL follow up (cHex/EtOAc 10:1, 2 runs) revealed that the starting 47 (R.sub.f 0.6) had been converted to a closely migrating product (R.sub.f 0.65). NIS (260 mg, 1.1 mmol, 1.1 equiv.) and H.sub.2O (12 mL) were added and after stirring for another 1 h at rt, 10% aq. Na.sub.2SO.sub.3 was added. The reaction mixture was concentrated and the aq. phase was extracted with DCM (30 mL) thrice. The combined organic layers were washed with brine (50 mL), dried over anhyd. Na.sub.2SO.sub.4, and concentrated under reduced pressure. The residue was purified by flash chromatography with cHex/EtOAc (80:20.fwdarw.75:25) to give the expected hemiacetal 49 (870 mg, 0.95 mmol, 90%) as a white solid. The / hemiacetal 49 had R.sub.f 0.4, 0.45 (Tol/EtOAc, 4:1). The isomer had .sup.1H NMR (CDCl.sub.3) 7.85-7.75 (m, 4H, H.sub.Ar), 7.52-7.46 (m, 3H, H.sub.Ar), 7.39-7.26 (m, 8H, H.sub.Ar), 7.21-7.10 (m, 2H, H.sub.Ar), 6.70 (d, J.sub.2,NH=9.2 Hz, NH.sub.B), 5.80 (d, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A), 5.24 (t, 1H, J.sub.1,2=3.6 Hz, H-1.sub.B), 5.19 (brs, 2H, CH.sub.2Bn-6), 4.83 (brs, 2H, CH.sub.2Nap), 4.74 (d, 1H, J.sub.4,5=2.0 Hz, H-5.sub.A), 4.42-4.33 (m, 3H, H-2.sub.A, H-4.sub.A, H-2.sub.B), 4.34 (d.sub.po, 1H, J=12.0 Hz, CH.sub.2Bn), 4.25 (d.sub.po, 1H, J=12.0 Hz, CH.sub.2Bn), 4.23 (dd.sub.po, J.sub.3,4=3.2 Hz, J.sub.2,3=10.4 Hz, H-3.sub.A), 4.15 (dd.sub.po, J.sub.3,4=2.4 Hz, H-4.sub.B), 4.09-4.03 (m, 2H, H-3.sub.B, H-5.sub.B), 3.15 (d, 1H, J.sub.1,OH=3.6 Hz, OH), 2.39 (s, 3H, CH.sub.3Ac), 2.38 (s, 3H, CH.sub.3Ac), 1.21 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3), 175.1 (2C, CO.sub.NAc), 168.8 (C-6.sub.A), 161.9 (CO.sub.NTCA), 135.1, 134.8, 133.2, 133.0 (C.sub.q,Ar), 129.0, 128.8, 128.5, 128.4, 128.2, 127.9, 127.7 (2C), 126.4, 126.1, 125.6, 125.3 (C.sub.Ar), 98.8 (C-1.sub.A, .sup.1J.sub.C,H=175 Hz), 92.4 (CCl.sub.3), 91.2 (C-1.sub.B, .sup.1J.sub.C,H=176 Hz), 76.9 (C-3.sub.B), 73.9 (C-5.sub.A), 73.6 (C-4.sub.A), 73.0 (C-3.sub.A), 72.6 (CH.sub.2Nap), 71.8 (CH.sub.2Bn), 67.5 (CH.sub.2Bn-6), 65.5 (C-4.sub.B), 64.7 (C-5.sub.B), 59.1 (C-2.sub.A), 50.6 (C-2.sub.B), 21.4 (CH.sub.3Ac), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.35H.sub.44Cl.sub.3N.sub.6O.sub.11, 1615.3589; found 1615.3596.

    [0549] (Benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-4-O-(2-naphthylmethyl)-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2,4,6-trideoxy-2-trichloroacetamido-/-D-galactopyranosyl (N-phenyl)trifluoroacetimidate (50) and 2-Trichloromethyl-[(Benzyl 3-O-benzyl-4-O-(2-naphthylmethyl)-2-deoxy-2-(N,N-diacetyl)amino--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-1,2,4,6-tetradeoxy--D-galactopyrano]-[2,1,d]-oxazoline (51). Hemiacetal 49 was dissolved in acetone (12 mL) and PTFACl (113 L, 713 mol, 1.3 equiv.) was added followed by Cs.sub.2CO.sub.3 (197 mg, 604 mol, 1.1 equiv.). After stirring at rt for 2 h, a TLC follow up (Tol/EtOAc 4:1) showed the complete conversion of the hemiacetal (R.sub.f 0.4) into a less polar compound (R.sub.f 0.9). The suspension was filtered over a pad of Celite, washed with acetone (5 mL) twice, and the filtrate was concentrated. The residue was purified by column chromatography (cHex/EtOAc 90:10.fwdarw.85:15, +1% Et.sub.3N) to give a 4:1 mix of the expected PTFA donor 50 and oxazoline 51 (480 mg, 281 mol, 80%) as a white solid. The isolated mix of 50 and 51 had R.sub.f 0.9 (Tol/EtOAc 4:1). .sup.1H NMR (major compound, CDCl.sub.3) 7.85-6.70 (m, 21H, H.sub.Ar), 6.59 (d, 1H, J=8.4 Hz, NH), 5.94 (bs, 1H, H-1.sub.B), 6.37 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A), 5.21 (bs, 2H, CH.sub.2Bn-6), 6.37 (d, 1H, CH.sub.2Nap), 6.37 (d, 1H, J=12.1 Hz, CH.sub.2Nap), 4.88-4.81 (m.sub.po, 2.5H), 4.75 (d, 1H, J=2.0 Hz, H-5.sub.A), 4.55 (ddd, 1H, H-2.sub.B), 4.41 (dd.sub.po, 1H, J.sub.2,3=10.5 Hz, H-2.sub.A), 4.39-4.33 (m, 2H, H-4.sub.A, CH.sub.2Bn), 4.25 (d, 1H, J=11.8 Hz, CH.sub.2Bn), 4.21 (dd.sub.po, 1H, J.sub.2,3=2.8 Hz, H-3.sub.A), 4.19 (do, 1H, H-4.sub.B), 4.13 (d, 1H, J.sub.2,3=11.0 Hz, J.sub.3,4=3.2 Hz, H-3.sub.B), 3.90 (brq, 1H, H-5.sub.B), 2.38 (s, 6H, CH.sub.3Ac), 1.25 (d, 3H, J.sub.5,6=6.2 Hz, H-6.sub.B). .sup.13C NMR (major isomer, CDCl.sub.3) 175.0 (CO.sub.NAc), 168.8, 168.7 (C-6.sub.A), 162.0 (CO.sub.NTCA), 142.9, 137.2, 135.0, 134.7, 133.2, 133.0 (C.sub.q,Ar), 128.9, 128.8, 128.7 (2C), 128.6, 128.4 (2C), 128.2 (2C), 128.0, 127.8, 127.7 (2C), 126.5, 126.4, 126.3, 126.2, 126.1, 126.0 (2C), 125.7, 124.9, 120.4, 119.2 (C.sub.Ar), 98.5 (C-1.sub.A), 93.7 (br, C-1.sub.B), 92.0 (CCl.sub.3), 76.3 (C-3.sub.B), 74.1 (C-5.sub.A), 73.5 (C-4.sub.A), 73.0 (C-3.sub.A), 72.6 (CH.sub.2Nap), 71.8 (CH.sub.2Bn), 67.6 (CH.sub.2Bn-6), 67.5 (C-5.sub.B), 64.6 (C-4.sub.B), 59.0 (C-2.sub.A), 49.9 (C-2.sub.B), 29.6 (CH.sub.3Ac), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.51H.sub.48Cl.sub.3F.sub.3N.sub.6O.sub.11, 1100.2737; found 1100.2729.

    [0550] Allyl (benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.3)-(benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (52). PTFACl (102 L, 642 mol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (117 mg, 543 mol, 1.1 equiv.) were added to a solution of hemiacetal 49 (230 mg, 252 mol, 1.0 equiv.) in acetone (8.0 mL). After stirring for 2 h at rt, the suspension was filtered over a pad of Celite and solids were washed with acetone (5 mL) thrice. The filtrate was concentrated under reduced pressure and the crude product was subjected to the next step.

    [0551] The crude mix of glycosyl donors 50 and 51 (252 mol theo., 1.1 equiv.) and acceptor 48 (184 mg, 227 mol, 1.0 equiv.) were co-evaporated with anhyd. toluene (5 mL) and then dried under high vacuum for 1 h. The dried mixture was dissolved in anhyd. DCM (8.0 mL) and stirred for 1 h with freshly activated MS 4 (500 mg) under an Ar atmosphere. The reaction mixture was cooled to 0 C. and TfOH (1.1 L, 13 mol, 0.05 equiv.) was added. After stirring for 30 min at this temperature, a TLC analysis (Tol/EtOAc, 4:1) showed no further evolution while donor 50/51 (R.sub.f 0.9) had reacted and a more polar spot (R.sub.f 0.35) was visible. Et.sub.3N (2.0 L) was added and the suspension was filtered over a fitted funnel. Solids were washed with DCM (5 mL) twice and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 80:20.fwdarw.60:40) to give firstly the condensation product 52 (240 mg, 141 mol, 62%; corr. yield 85%) as a white solid and then some unreacted 48 (50 mg, 7%). Tetrasaccharide 52 had .sup.1H NMR (CDCl.sub.3) 7.82-7.74 (m, 4H, H.sub.Ar), 7.51-7.10 (m, 23H, H.sub.Ar), 6.99 (d, J.sub.2,NH=6.8 Hz, NH.sub.B1), 6.74 (d, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.90 (m, 1H, CH.sub.All), 5.78 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A1), 5.65 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A), 5.29-5.15 (m, 6H, CH.sub.2All, CH.sub.2Bn-6), 5.00 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B1), 4.85-4.79 (m, 3H, CH.sub.2Nap, H-5.sub.A1), 4.76 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B), 4.72 (d, 1H, J.sub.4,5=2.4 Hz, H-5.sub.A), 4.64 (dd, 1H, J.sub.3,4=4.0 Hz, J.sub.2,3=10.8 Hz, H-3.sub.B), 4.45-4.22 (m, 11H, H-2.sub.A1, H-3.sub.B1, H-4.sub.A, H-4.sub.A1, H-3.sub.A, H-3.sub.A1, CH.sub.2All, CH.sub.2Bn), 4.07-3.99 (m, 4H, CH.sub.2All, H-2.sub.A, H-4.sub.B1, H-4.sub.B), 3.54-3.41 (m, 4H, H-2.sub.B, H-2.sub.B1, H-5.sub.B, H-5.sub.B1), 2.38 (brs, 12H, CH.sub.3Ac), 2.23 (brs, 3H, CH.sub.3Ac), 1.28 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B), 1.19 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B1). .sup.13C NMR (CDCl.sub.3) 175.3-174.8 (br, 4C, CO.sub.NAc), 168.7, 168.3 (C-6.sub.A, C-6.sub.A1), 161.9, 161.8 (2CO.sub.NTCA), 137.4, 135.2, 135.1, 135.0, 133.2, 133.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 129.0, 128.9, 128.8, 128.7, 128.6, 128.5, 128.4, 128.2, 128.1, 128.0, 127.9, 127.8, 127.7, 127.6, 126.3, 126.1, 125.9, 125.6, 125.2 (C.sub.Ar), 117.9 (CH.sub.2All), 98.9 (C-1.sub.A1*, .sup.1J.sub.C,H=175 Hz), 98.8 (C-1.sub.B1, .sup.1J.sub.C,H=167 Hz), 98.3 (C-1.sub.A*, .sup.1J.sub.C,H=175 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 92.2, 91.8 (CCl.sub.3), 76.7 (C-3.sub.B1), 76.2 (C-5.sub.A1), 75.9 (C-3.sub.B), 73.4 (C-5.sub.A), 73.6 (C-4.sub.A), 72.8, 72.6 (C-3.sub.A, C-3.sub.A1), 72.5 (CH.sub.2Nap), 72.0, 71.8 (CH.sub.2Bn), 71.4 (C-4.sub.A1), 70.1 (CH.sub.2All), 68.7, 68.6 (C-5.sub.B, C-5.sub.B1), 67.5 (2C, CH.sub.2Bn-6), 65.3 (C-4.sub.B), 65.2 (C-4.sub.B1), 59.3 (C-2.sub.A), 59.1 (C-2.sub.A1), 58.8 (C-2.sub.B1), 55.3 (C-2.sub.B), 27.7, 25.3, 21.4 (4C, CH.sub.3Ac), 17.4, 17.2 (C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.78H.sub.86Cl.sub.6N.sub.6O.sub.21 1722.4131; found 1722.4110.

    [0552] Allyl (benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.3)-(benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (53). DDQ (108 mg, 475 mol, 3.0 equiv.) was added to tetrasaccharide 52 (270 mg, 158 mol, 1.0 equiv.) in DCM/phosphate buffer pH 7 (8:1, 18 mL) cooled to 0 C. The biphasic mixture was stirred vigorously for 4 h while allowing the bath to slowly warm to rt. At completion, 10% aq. NaHCO.sub.3 (10 mL) was added followed by DCM (20 mL). The DCM layer was separated, washed with water and brine, dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The crude was purified by flash chromatography with Tol/EtOAc (75:25.fwdarw.70:30). Alcohol 53 (180 mg, 115 mol, 73%), obtained as a white solid, had R.sub.f 0.3 (Tol/EtOAc, 6:4). .sup.1H NMR (CDCl.sub.3) 7.44-7.13 (m, 20H, H.sub.Ar), 6.97 (d, J.sub.2,NH=6.8 Hz, NH.sub.B1), 6.80 (d, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.88 (m, 1H, CH.sub.All), 5.68 (d, 1H, J.sub.1,2=7.8 Hz, H-1.sub.A1), 5.64 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A), 5.34-5.15 (m, 6H, CH.sub.2All, CH.sub.2Bn-6), 4.95 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B1), 4.79 (d, 1H, J.sub.4,5=2.0 Hz, H-5.sub.A1), 4.76 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.72 (d, 1H, J.sub.4,5=2.4 Hz, H-5.sub.A), 4.64 (dd, 1H, J.sub.3,4=4.0 Hz, J.sub.2,3=10.4 Hz, H-3.sub.B), 4.50-4.37 (m, 7H, H-3.sub.B1, H-4.sub.A, H-4.sub.A1, H-3.sub.A, H-3.sub.A1, CH.sub.2Bn), 4.32-4.24 (m, 3H, CH.sub.2Bn, CH.sub.2All), 4.13-3.99 (m, 4H, CH.sub.2All, H-2.sub.A1, H-2.sub.A, H-4.sub.B1, H-4.sub.B), 3.54-3.41 (m, 4H, H-2.sub.B, H-2.sub.B1, H-5.sub.B, H-5.sub.B1), 2.61 (d, 1H, J.sub.4,OH=2.0 Hz, OH), 2.38-2.32 (brs, 9H, CH.sub.3Ac), 2.22 (brs, 3H, CH.sub.3Ac), 1.28 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B), 1.19 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B1). .sup.13C NMR (CDCl.sub.3) 175.3, 174.7 (CO.sub.NAc), 168.3, 168.2 (C-6.sub.A), 161.9, 161.8 (CO.sub.NTCA), 137.4, 136.9, 135.0 (2C) (C.sub.q,Ar), 133.5 (CH.sub.All), 129.0, 128.9, 128.7 (2C), 128.6 (2C), 128.5, 128.4, 128.3, 128.2, 128.0 (2C), 125.2 (C.sub.Ar), 117.9 (CH.sub.2All), 98.9 (C-1.sub.B1, .sup.1J.sub.C,H=168 Hz), 98.4 (C-1.sub.A, .sup.1J.sub.C,H=176 Hz), 98.4 (C-1.sub.A1, .sup.1J.sub.C,H=177 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.2, 91.9 (2C, CCl.sub.3), 76.7 (C-3.sub.B1), 76.2 (C-5.sub.A1), 75.7 (C-3.sub.B), 75.2 (C-5.sub.A), 72.6 (C-4.sub.A), 72.5, 71.4 (C-3.sub.A, C-3.sub.A1), 72.3, 72.0 (2C, CH.sub.2Bn), 70.1 (CH.sub.2All), 68.7, 68.6 (C-5.sub.B, C-5.sub.B1), 67.5, 67.5 (2C, CH.sub.2Bn-6), 66.5 (C-4.sub.A1), 65.3 (2C, C-4.sub.B, C-4.sub.B1), 59.3 (C-2.sub.A), 59.1 (C-2.sub.A1), 55.7 (C-2.sub.B1), 55.2 (C-2.sub.B), 27.7, 25.3, 21.4 (4C, CH.sub.3Ac), 17.4, 17.2 (C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.67H.sub.78Cl.sub.6N.sub.11O.sub.21 1582.3505; found 1582.3503.

    [0553] Allyl (benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.3)-(benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.3)-(benzyl 3-O-benzyl-2-(N,N-diacetyl)amino-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (54). Hemiacetal 49 (131 mg, 144 mol, 1.0 equiv.) was dissolved in acetone (7.0 mL). PTFACl (30 L, 187 mol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (52 mg, 158 mol, 1.1 equiv.) were added and the mixture stirred at rt for 2 h. Solids were filtered off over a pad of Celite and washed with acetone (5 mL) twice. The filtrate was concentrated under reduced pressure and the crude donor, isolated as a 4:1 mix of PTFA 50 and oxazoline 51, was subjected to the next step without further purification.

    [0554] The crude mix of donors 50/51 (144 mol theo., 1.25 equiv.) and acceptor 53 (180 mg, 115 mol, 1.0 equiv.) were coevaporated with anhyd. toluene (5 mL) twice and then dried extensively under high vacuum. The mixture, dissolved in anhyd. DCM (6.0 mL), was stirred with freshly activated MS 4 (200 mg) for 30 min at rt under an Ar atmosphere, and cooled to 0 C. TfOH (1.0 L, 0.05 equiv.) was added and after stirring for 30 min at this temperature, a TLC analysis indicated donor consumption and the presence of a major new product together with some unreacted acceptor. Et.sub.3N (2.0 L) was added and after 10 min, solids were filtered off and washed with DCM (5 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/EA 75:25.fwdarw.60:40) to give first the condensation product 54 (200 mg, 81 mol, 71%; corr. yield 98%) as a white solid, followed by some unreacted 53 (50 mg, 28%). Hexasaccharide 54 had R.sub.f 0.25 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.85-7.11 (m, 37H, H.sub.Ar), 6.94 (d, J.sub.2,NH=6.8 Hz, NH.sub.B2*), 6.90 (d, J.sub.2,NH=6.8 Hz, NH.sub.B1*), 6.74 (d, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.91-5.81 (m, 1H, CH.sub.All), 5.78 (d, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A2), 5.65 (d, 1H, J.sub.1,2=7.8 Hz, H-1.sub.A1*), 5.64 (d, 1H, J.sub.1,2=7.8 Hz, H-1.sub.A*), 5.31-5.16 (m, 8H, CH.sub.2All, 3CH.sub.2Bn-6), 5.03 (d, 1H, J.sub.1,2=8.1 Hz, H-1.sub.B2*), 4.99 (d, 1H, J.sub.1,2=8.1 Hz, H-1.sub.B1*), 4.82-4.80 (m, 2H, CH.sub.2Nap), 4.80 (d.sub.po, 1H, J.sub.4,5=2.2 Hz, H-5.sub.A*) 4.78 (d.sub.po, 1H, J.sub.4,5=2.4 Hz, H-5.sub.A1*), 4.76 (d.sub.po, 1H, J.sub.1,2=8.3 Hz, H-1.sub.B), 4.72 (d, 1H, J.sub.4,5=2.3 Hz, H-5.sub.A2*), 4.68 (dd, 1H, J.sub.3,4=4.0 Hz, J.sub.2,3=10.7 Hz, H-3.sub.B2*), 4.59 (dd, 1H, J.sub.3,4=4.0 Hz, J.sub.2,3=10.6 Hz, H-3.sub.B1*), 4.44 (dd, 1H, J.sub.3,4=3.8 Hz, J.sub.2,3=10.7 Hz, H-3.sub.B), 4.41-4.22 (m, 15H, H-2.sub.A2, H-3.sub.B, H-4.sub.A, H-4.sub.A1, H-4.sub.A2, H-3.sub.A, H-3.sub.A1, H-3.sub.A2, CH.sub.2All, 3CH.sub.2Bn), 4.09 (bd.sub.po, 1H, H-4.sub.B1*), 4.07 (bd.sub.po, 1H, H-4.sub.B), 4.05-3.99 (m, 4H, CH.sub.2All, H-2.sub.A, H-2.sub.A1, H-4.sub.B2*), 3.52 (dt, 1H, H-2.sub.B), 3.50-3.43 (m, 3H, H-5.sub.B, H-5.sub.B1, H-5.sub.B2), 3.42-3.36 (m, 2H, H-2.sub.B1, H-2.sub.B2), 2.41 (brs, 12H, 4CH.sub.3Ac), 2.23 (brs, 6H, 2CH.sub.3Ac), 1.30 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B*), 1.23 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B1*) 1.12 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B2*). .sup.13C NMR (CDCl.sub.3) 175.6, 175.3, 175.0, 174.7 (br, 6C, CO.sub.NAc), 168.9, 168.8, 168.7 (3C, C-6.sub.A), 162.3, 161.8, 161.7 (3C, CO.sub.NTCA), 137.5, 137.4, 135.2, 135.1, 135.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 133.2, 133.0 (C.sub.q,Ar), 129.0, 128.9, 128.8, 128.7, 128.6, 128.5, 128.4, 128.2, 128.1, 128.0, 127.8, 127.7, 126.4, 126.1, 125.9, 125.6, 125.2 (27C, C.sub.Ar), 117.9 (CH.sub.2All), 98.9 (C-1.sub.B2*, .sup.1J.sub.C,H=167 Hz), 98.8 (C-1.sub.A2, .sup.1J.sub.C,H=175 Hz), 98.7 (C-1.sub.B1*, .sup.1J.sub.C,H=167 Hz), 98.4 (C-1.sub.A1*, .sup.1J.sub.C,H=177 Hz), 98.3 (C-1.sub.A*, .sup.1J.sub.C,H=177 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 76.7, 75.7, 75.1 (C-3.sub.B, C-3.sub.B1, C-3.sub.B2), 76.2, 74.0 (3C, C-5.sub.A, C-5.sub.A1, C-5.sub.A2), 73.7, 71.3 (3C, C-4.sub.A, C-4.sub.A1, C-4.sub.A2), 72.8, 72.4, 70.9 (C-3.sub.A, C-3.sub.A1, C-3.sub.A2), 72.5 (CH.sub.2Nap), 72.0, 71.9 (3C, CH.sub.2Bn), 70.1 (CH.sub.2All), 68.6, 68.5 (C-5.sub.B, C-5.sub.B1, C-5.sub.B2), 67.5 (3C, CH.sub.2Bn-6), 65.3, 65.2 (C-4.sub.B, C-4.sub.B1, C-4.sub.B2), 59.5, 59.4, 59.1 (C-2.sub.A, C-2.sub.A1, C-2.sub.A2), 55.9, 55.2 (C-2.sub.B, C-2.sub.B1, C-2.sub.B2), 27.7, 25.4 (3C, CH.sub.3Ac), 17.4, 17.3 (C-6.sub.B, C-6.sub.B1, C-6.sub.B2). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.110H.sub.124Cl.sub.9N.sub.17O.sub.31 1246.7922; found 1246.7922.

    [0555] Allyl (benzyl 3-O-benzyl-2-deoxy-2-(N,N-diacetyl)amino--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.3)-(benzyl 3-O-benzyl-2-deoxy-2-(N,N-diacetyl)amino--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.3)-(benzyl 3-O-benzyl-2-deoxy-2-(N,N-diacetyl)amino--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (55). DDQ (55 mg, 244 mol, 3.0 equiv.) was added to hexasaccharide 54 (200 mg, 81 mol, 1.0 equiv.) in DCM (8.0 mL) and phosphate buffer pH 7 (1.0 mL). The biphasic mixture was cooled to 0 C. and stirred for 2 h. Additional DDQ (200 mg, 81 mol, 1.0 equiv.) was added and stirring was pursued for another 4 h while the bath temperature reached rt. A TLC analysis (Tol/EtOAC 3:1) showed the absence of the fully protected 54 (R.sub.f 0.6) and the presence of a more polar spot (R.sub.f 0.4). 10% Aq. NaHCO.sub.3 (5 mL) was added followed by DCM (15 mL). The DCM layer was separated, washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 4:1.fwdarw.3:1) to give alcohol 55 (120 mg, 51 mol, 64%) as a white solid. Hexasaccharide 55 had .sup.1H NMR (CDCl.sub.3) 7.36-7.13 (m, 30H, H.sub.Ar), 6.89 (d.sub.po, J.sub.2,NH=6.8 Hz, NH.sub.B*), 6.87 (d, J.sub.2,NH=6.8 Hz, NH.sub.B1*), 6.72 (d, J.sub.2,NH=7.6 Hz, NH.sub.B2*), 5.90-5.80 (m, 1H, CH.sub.All), 5.68 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A*), 5.65 (d.sub.po, 1H, J.sub.1,2=7.8 Hz, H-1.sub.A1*), 5.65 (d.sub.po, 1H, J.sub.1,2=7.8 Hz, H-1.sub.A2*), 5.35-5.15 (m, 8H, CH.sub.2All, 3CH.sub.2Bn-6), 5.01 (d.sub.po, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B*), 4.94 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B1*), 4.79-4.75 (m, 6H, CH.sub.2Nap, H-5.sub.A*, H-5.sub.A1*, H-3.sub.B*, H-1.sub.B2*), 4.72 (d, 1H, J.sub.4,5=2.4 Hz, H-5.sub.A2*), 4.69 (dd, 1H, J.sub.3,4=4.0 Hz, J.sub.2,3=10.8 Hz, H-3.sub.B1*), 4.58 (dd, 1H, J.sub.3,4=4.0 Hz, J.sub.2,3=10.4 Hz, H-3.sub.B2*), 4.45-4.23 (m, 15H, H-3.sub.B, H-4.sub.A, H-4.sub.A1, H-4.sub.A2, H-3.sub.A, H-3.sub.A1, H-3.sub.A2, CH.sub.2All, 3CH.sub.2Bn), 4.08-3.99 (m, 7H, H-2.sub.A, H-2.sub.A1, H-2.sub.A2, H-4.sub.B, H-4.sub.B1, H-4.sub.B2, CH.sub.2All), 3.54-3.35 (m, 6H, H-2.sub.B, H-2.sub.B1, H-2.sub.B2, H-5.sub.B, H-5.sub.B1, H-5.sub.B2), 2.52 (d, 1H, J.sub.4,OH=2.4 Hz, OH), 2.40-2.17 (brm, 18H, CH.sub.3Ac), 1.29-1.12 (m, 12H, H-6.sub.B, H-6.sub.B1, H-6.sub.B2). .sup.13C NMR (CDCl.sub.3) 175.3 (6C, CO.sub.NAc), 168.3 (2C), 168.1 (3C, C-6.sub.A), 161.8 (2C), 161.7 (3C, CO.sub.NTCA), 133.5, 133.5 (CH.sub.All), 137.4, 136.8, 135.0 (2C), 127.8, 127.0 (C.sub.q,Ar), 129.0, 129.4, 128.8 (2C), 128.7 (2C), 128.6 (2C), 128.5 (2C), 128.4 (2C), 128.3, 128.2, 128.0 (2C), 125.2 (30C, C.sub.Ar), 117.9 (CH.sub.2All), 98.9 (C-1.sub.B*, .sup.1J.sub.C,H=166 Hz), 98.7 (C-1.sub.B1*, .sup.1J.sub.C,H=167 Hz), 98.4 (C-1.sub.A, .sup.1J.sub.C,H=175 Hz), 98.3 (2C, C-1.sub.A1*, C-1.sub.A2*, .sup.1J.sub.C,H=175 Hz), 97.6 (C-1.sub.B2*, .sup.1J.sub.C,H=164 Hz), 92.6, 91.8 (3C, CCl.sub.3), 76.6, 75.5, 75.1 (C-3.sub.B, C-3.sub.B1, C-3.sub.B2), 76.2 (2C), 75.2 (C-5.sub.A, C-5.sub.A1, C-5.sub.A2), 72.8, 72.6, 72.4 (C-3.sub.A, C-3.sub.A1, C-3.sub.A2), 72.3, 72.0 (3C, 3CH.sub.2Bn), 71.3, 71.0 (2C, C-4.sub.A, C-4.sub.A1), 70.1 (CH.sub.2All), 68.6 (2C), 68.5 (C-5.sub.B, C-5.sub.B1, C-5.sub.B2), 67.7, 67.5 (3C, CH.sub.2Bn-6), 66.5 (C-4.sub.A2), 65.4, 65.3, 65.2 (C-4.sub.B, C-4.sub.B1, C-4.sub.B2), 59.5, 59.3, 59.0 (C-2.sub.A, C-2.sub.A1, C-2.sub.A2), 55.8 (2C), 55.3 (C-2.sub.B, C-2.sub.B1, C-2.sub.B2), 27.7 (6C, CH.sub.3Ac), 17.4, 17.3, 17.2 (3C, C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.99H.sub.112C.sub.19N.sub.16O.sub.31 2335.4876; found 2335.4871.

    [0556] Allyl (benzyl 3-O-benzyl-2-deoxy-2-(N,N-diacetyl)amino-4-O-(2-naphthylmethyl)--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.3)-(benzyl 3-O-benzyl-2-deoxy-2-(N,N-diacetyl)amino--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.3)-(benzyl 3-O-benzyl-2-deoxy-2-(N,N-diacetyl)amino--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.3)-(benzyl 3-O-benzyl-2-deoxy-2-(N,N-diacetyl)amino--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (56). A 4:1 mix of donors 50/51 (92 mg, 85 mol, 1.5 equiv.) and acceptor 55 (132 mg, 57 mol, 1.0 equiv.) was coevaporated with toluene (3 mL) twice and dried extensively under high vacuum. The mixture, dissolved in anhyd. DCM (4.0 mL), was stirred for 45 min at rt with freshly activated MS 4 and cooled to 0 C. TfOH (3.8 L, 0.05 equiv.) was added and the reaction mixture was stirred at this temperature. At completion, as revealed by TLC analysis (Tol/EtOAc 7:3), Et.sub.3N (5.0 L) was added and after 10 min, solids were filtered over a fitted funnel. The filtrate was concentrated to dryness and the residue was purified by flash chromatography (Tol/EtOAc 80:20.fwdarw.70:30) to give first the glycosylation product 56 (60 mg, 19 mol, 33%, corr. yield, 54% wrt acceptor) as a white solid followed by the remaining unreacted acceptor 55 (50 mg, 38%). Octasaccharide 56 had R.sub.f 0.35 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.85-7.74 (m, 5H, H.sub.Ar), 7.52-7.11 (m, 42H, H.sub.Ar), 6.90-6.86 (m, 3H, NH.sub.B1, NH.sub.B3, NH.sub.B3), 6.71 (d, 1H, J.sub.2,NH=7.3 Hz, NH.sub.B), 5.90-5.75 (m, 1H, CH.sub.All), 5.78 (d, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A3), 5.66-5.62 (m, 3H, H-1.sub.A, H-1.sub.A1, H-1.sub.A2), 5.29-5.16 (m, 10H, CH.sub.2All, 4CH.sub.2Bn-6), 5.03-5.01 (m, 2H, H-1.sub.B1, H-1.sub.B2), 4.96 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B3), 4.83-4.80 (m, 3H, CH.sub.2Nap, H-5.sub.A), 4.78-4.76 (m, 3H, H-1.sub.B, H-5.sub.A1*, H-5.sub.A2*), 4.72-4.55 (m, 4H, H-3.sub.B, H-3.sub.B1, H-3.sub.B2, H-5.sub.A3*), 4.45-4.22 (m, 17H, H-2.sub.A*, H-3.sub.B3*, H-3.sub.A, H-3.sub.A1, H-3.sub.A2, H-3.sub.A3, H-4.sub.A, H-4.sub.A1, H-4.sub.A2, H-4.sub.A3, CH.sub.2All, 3CH.sub.2Bn), 4.09-3.98 (m, 8H, CH.sub.2All, H-2.sub.A1, H-2.sub.A2, H-2.sub.A3, 4H-4.sub.B), 3.55-3.32 (m, 8H, 4H-2.sub.B, 4H-5.sub.B), 2.40-2.19 (m.sub.po, 24H, 8CH.sub.3NAc), 1.30-1.18 (d, 12H, J.sub.5,6=6.4 Hz, H-6.sub.B, H-6.sub.B1, H-6.sub.B2, H-6.sub.B3). .sup.13C NMR (Partial, CDCl.sub.3) 175.5, 175.0, 174.6 (CO.sub.NAc), 168.7 (2C), 168.3, 168.2 (C-6.sub.A), 162.3, 161.8, 161.7 (4C, CO.sub.NTCA), 137.4 (2C) 137.3, 135.2 (2C), 135.1, 135.0 (2C), 133.2, 133.0, 126.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 129.0, 128.9, 128.8, 128.7 (2C), 128.6, 128.5 (2C), 128.4 (2C), 128.2, 128.1, 128.0, 127.8, 127.7, 126.4, 126.1, 125.9, 125.6, 125.2 (C.sub.Ar), 117.9 (CH.sub.2All), 98.9 (C-1.sub.A3*, .sup.1J.sub.C,H=178 Hz), 98.8 (C-1.sub.B3*, .sup.1J.sub.C,H=166 Hz), 98.7 (C-1.sub.B1, C-1.sub.B2, .sup.1J.sub.C,H=168 Hz), 98.4 (C-1.sub.A1*, C-1.sub.A2*, J.sub.C,H=178 Hz), 98.3 (C-1.sub.A*, .sup.1J.sub.C,H=178 Hz), 97.6 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.2, 91.8 (4C, CCl.sub.3), 76.6, 75.8, 75.1, 74.9 (4C, C-3.sub.B, C-3.sub.B1, C-3.sub.B2, C-3.sub.B3), 76.2, 73.7 (4C, C-5.sub.A, C-5.sub.A1, C-5.sub.A2, C-5.sub.A3), 74.0, 72.9, 72.8 (4C, C-4.sub.A, C-4.sub.A1, C-4.sub.A2, C-4.sub.A3), 72.5 (CH.sub.2Nap), 71.8, 70.9 (3C, C-3.sub.A, C-3.sub.A1, C-3.sub.A2), 72.0, 71.9 (4C, CH.sub.2Bn), 70.1 (CH.sub.2All), 68.6, 68.5 (4C, C-5.sub.B, C-5.sub.B1, C-5.sub.B2, C-5.sub.B3), 67.5 (4C, CH.sub.2Bn-6), 65.4 (4C, C-4.sub.B, C-4.sub.B1, C-432, C-43), 59.5, 59.4, 59.1 (4C, C-2.sub.A, C-2.sub.A1, C-2.sub.A2, C-2.sub.A3), 55.8, 55.3 (4C, C-2.sub.B, C-2.sub.B1, C-2.sub.B2, C-2.sub.B3), 27.6, 25.2, 12.4 (8C, CH.sub.3Ac), 17.4, 17.3, 17.2 (4C, C-6.sub.B, C-4.sub.B1, C-4.sub.B2, C-4.sub.B3). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.142H.sub.154Cl.sub.12N.sub.21O.sub.41 3229.6899; found 3230.7006.

    [0557] Full Deprotection

    ##STR00094##

    [0558] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (1)..sup.[1] 20% Pd(OH).sub.2/C (100 mg) was added to a solution of 47 (50 mg, 53 mol, 1.0 equiv.) in .sup.tBuOH/DCM/H.sub.2O (17 mL, 20:5:2, v/v/v). After stirring for 48 h in a hydrogen atmosphere. The suspension was passed through a 0.2 m filter and washed thoroughly with methanol. The filtrate was concentrated and the crude product was dried under vacuum. The obtained white powder was dissolved in methanol (5 mL) and hydroxylamine (3.7 mg, mol, 1.0 equiv.) was added. Monitoring by LCMS revealed the full consumption of the mono-acetate product and the presence of the desired product (LCMS: [M+H].sup.+ m/z 867.2) after 4 h. Phosphate buffer pH 7 was added with frequent pH monitoring to achieve pH 7. The mixture was diluted with water (10 mL) and lyophilized. After freeze-drying, purification of the crude material by semi-preparative RP-HPLC gave propyl glycoside 1 as a white solid (14 mg, 30 mol, 57%). Disaccharide 1 had RP-HPLC (215 nm) R.sub.t=13.9 min (conditions B), R.sub.t=12.2 min (conditions C). HRMS (ESI.sup.+): m/z calcd for C.sub.19H.sub.35N.sub.3O.sub.9Na [M+Na].sup.+ m/z 486.2064; found 486.2067. NMR data were as published..sup.[1].

    [0559] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (2). Tetrasaccharide 54 (30 mg, 18 mol, 1.0 equiv.) was dissolved in .sup.tBuOH/DCM/H.sub.2O (10 mL, 20:5:2, v/v/v) and 20% Pd(OH).sub.2/C (100 mg) was added. The reaction mixture was degassed several times and stirred under a hydrogen atmosphere for 48 h. A follow up by HRMS revealed the presence of a major product corresponding the 2.sub.A-NAc.sub.2,2.sub.B-NAc product (HRMS: C.sub.39H.sub.62N.sub.6O.sub.21Na [M+Na].sup.+ m/z 973.4270). The suspension was passed through a syringe filter (0.2 m) and washed thoroughly with methanol. The filtrate was concentrated and the crude product was dried under vacuum. The resulting white powder was dissolved in methanol (3.0 mL) and hydroxylamine (2.2 mg, 36 mol, 2.0 equiv.) was added. LCMS monitoring revealed that after 4 h, no intermediate remained and the desired product (LCMS: [M+H].sup.+ m/z 867.3) was present to a large extent. The reaction mixture was neutralized with phosphate buffer with frequent pH monitoring to achieve pH 7, then diluted with water (6.0 mL) and lyophilized. Purification of the crude material by semi-preparative RP-HPLC gave the propyl glycoside 2 as a white solid (5.9 mg, 6.8 mol, 39%). Tetrasaccharide 2 had RP-HPLC (215 nm) R.sub.t=12.3 min (conditions A), R.sub.t=13.6 min (conditions B), .sup.1H NMR (D.sub.2O) 4.87 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A), 4.77 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A1), 4.74 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-11), 4.66 (brs, 1H, H-5.sub.A), 4.60 (brs, 1H, H-5.sub.A1), 4.44 (d.sub.po, 2H, J=8.4 Hz, H-1.sub.B, H-4.sub.A), 4.36 (brs, 1H, H-4.sub.A1), 4.15-4.08 (m, 2H, H-3.sub.B, H-3.sub.B1), 4.03-3.99 (m.sub.po, 2H, H-5.sub.B, H-5.sub.B1), 3.96-3.90 (m, 2H, H-2.sub.A, H-4.sub.B), 3.80-3.66 (m, 7H, H-3.sub.A, H-3.sub.A1, H-2.sub.A1, H-2.sub.B, H-2.sub.B1, H-41, OCH.sub.2Pr), 3.51-3.49 (m.sub.po, 1H, OCH.sub.2Pr), 1.99, 1.94 (2s, 12H, CH.sub.3Ac), 1.51-1.48 (m, 2H, CH.sub.2Pr), 1.29 (d.sub.po, 6H, H-6.sub.B, H-6.sub.B1), 0.82 (t, 3H, J=7.2 Hz, CH.sub.3Pr). .sup.13C NMR (D.sub.2O) 174.6, 174.5 (2C), 174.0 (4C, CO.sub.NHAc), 172.7, 172.5 (2C, C-6.sub.A, C-6.sub.A1), 102.9 (C-1.sub.B1, .sup.1J.sub.C,H=168 Hz), 101.6 (C-1.sub.B, .sup.1J.sub.C,H=166 Hz), 101.1 (2C, C-1.sub.A, C-1.sub.A1, .sup.1J.sub.C,H=170 Hz, .sup.1J.sub.C,H=168 Hz), 76.7 (C-4.sub.A), 76.5, 76.3, 76.0 (4C, C-5.sub.A, C-5.sub.A1, C-3.sub.B, C-3.sub.B1), 72.7 (OCH.sub.2Pr), 68.5 (C-4.sub.A), 67.8, 67.6 (2C, C-3.sub.A, C-3.sub.A1), 67.4, 67.3 (2C, C-5.sub.B, C-5.sub.B1), 54.8, 54.7 (2C, C-4.sub.B, C-4.sub.B1), 51.5 (C-2.sub.A1), 51.3 (C-2.sub.A), 51.0, 50.8 (2C, C-2.sub.B, C-2.sub.B1), 23.3, 22.2 (4C, CH.sub.3Ac), 21.2 (CH.sub.2Pr), 15.6, 15.5 (2C, C-6.sub.B, C-6.sub.B), 9.5 (CH.sub.3Pr). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.35H.sub.58N.sub.6O.sub.19Na, 889.3649; found 889.3636.

    [0560] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (3). Hexasaccharide 54 (70 mg, 31 mol, 1.0 equiv.) was dissolved in .sup.tBuOH/DCM/H.sub.2O (19 mL, 20:5:2, v/v/v). 20% Pd(OH).sub.2/C (120 mg) was added and the suspension was degassed repeatedly. After stirring under a hydrogen atmosphere for 48 h, monitoring by LCMS analysis showed the presence of the targeted intermediate (LCMS: [M+H].sup.+ m/z 1396.4). The suspension was passed through a 0.2 m filter and washed extensively with methanol. The filtrate was concentrated and the crude material was dried under vacuum for 2 h. The obtained white powder was dissolved in methanol (3.0 mL) and hydroxylamine (6.1 mg, 85 mol, 3.0 equiv.) was added. After stirring for 3 h, LCMS analysis showed the complete disappearance of the triacetate intermediate and the presence of the desired product (LCMS: [M+H].sup.2+ m/z 635.2). Water (6.0 mL) was added and the reaction mixture was lyophilized. Purification of the crude material by semi-preparative RP-HPLC gave hexasaccharide 3 as a white solid (12 mg, 9.4 mol, 31%). The propyl glycoside 3 had RP-HPLC (215 nm) R.sub.t=11.3 min (conditions A), R.sub.t=13.3 min (conditions B). .sup.1H NMR (D.sub.2O) 4.88 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A2), 4.78 (d.sub.po, 2H, J.sub.1,2=8.4 Hz, H-1.sub.A, H-1.sub.A1), 4.75 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B1, H-1.sub.B2), 4.70 (brs.sub.po, 2H, H-5.sub.A, H-5.sub.A1), 4.61 (brs, 1H, H-5.sub.A2), 4.62 (m.sub.po, 2H, H-1.sub.B, H-4.sub.A), 4.36 (brs, 1H, H-4.sub.A1), 4.16-4.09 (m, 3H, H-3.sub.B, H-3.sub.B1, H-3.sub.B2), 4.06-3.90 (m, 7H, H-5.sub.B, H-5.sub.B1, H-5.sub.B2, H-2.sub.A, H-2.sub.A1, H-4.sub.A, H-4.sub.B), 3.86-3.72 (m, 10H, H-3.sub.A, H-3.sub.A1, H-3.sub.A2, H-4.sub.B1, H-4.sub.B2, H-2.sub.A2, H-2.sub.B, H-2.sub.B1, H-2.sub.B2, OCH.sub.2Pr), 3.51-3.49 (m.sub.po, 1H, OCH.sub.2Pr), 1.99, 1.98, 1.96, 1.95 (4 s, 18H, CH.sub.3Ac), 1.54-1.45 (m, 2H, CH.sub.2Pr), 1.30-1.29 (d.sub.po, 9H, H-6.sub.B), 0.82 (t, 3H, J=7.2 Hz, CH.sub.3Pr). .sup.13C NMR (D.sub.2O) 174.7, 174.5 (2C), 174.4 (2C), 174.0 (6C, CO.sub.NAcA,B), 172.6, 172.2 (2C) (3C, C-6.sub.A, C-6.sub.A1, C-6.sub.A2), 102.9 (2C, C-1.sub.B1, C-1.sub.B2, .sup.1J.sub.C,H=174.2 Hz, .sup.1J.sub.C,H=170.2 Hz), 101.6 (C-1.sub.B, .sup.1J.sub.C,H=162.0 Hz), 101.2, 101.1 (3C, C-1.sub.A, C-1.sub.A1, C-1.sub.A2), 76.6 (2C, C-4.sub.A, C-4.sub.A1), 76.3, 76.1 (3C, C-5.sub.A, C-5.sub.A1, C-5.sub.A2), 75.7 (3C, C-3.sub.B, C-3.sub.B1, C-3.sub.B2), 72.7 (OCH.sub.2Pr), 68.4 (C-4.sub.A2), 68.2, 67.8 (3C, C-3.sub.A, C-3.sub.A1, C-3.sub.A2), 67.6, 67.3 (3C, C-5.sub.B, C-51, C-5.sub.B2), 54.8, 54.7 (3C, C-4.sub.B, C-4.sub.B1, C-4.sub.B2), 51.5, 51.3 (3C, C-2.sub.A, C-2.sub.A1, C-2.sub.A2), 50.9, 50.8 (3C, C-2.sub.B, C-2.sub.B1, C-2.sub.B2), 23.3, 22.2 (6C, CH.sub.3Ac), 21.2 (CH.sub.2Pr), 15.6 (3C, C-6.sub.B, C-6.sub.B1, C-6.sub.B2), 9.5 (CH.sub.3Pr). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.51H.sub.85N.sub.9O.sub.28 635.7747; found 635.7736.

    [0561] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (4). Octasaccharide 56 (20 mg, 12 mol, 1.0 equiv.) was dissolved in .sup.tBuOH/DCM/H.sub.2O (16.5 mL, 20:5:2, v/v/v). 20% Pd(OH).sub.2/C (50 mg) was added and the suspension was degassed repeatedly. After stirring under a hydrogen atmosphere for 48 h, the suspension was passed through a 0.2 m filter and washed extensively with methanol. The filtrate was concentrated and the crude material was dried under vacuum for 2 h. HRMS analysis of the crude material showed the presence of the tetra-2-N-acetylacetamido product ([M+2H].sup.2+ calculated for: C.sub.75H.sub.118N.sub.12O.sub.41 921.3758, found 921.3753). The obtained white powder was dissolved in methanol (3.0 mL) and hydroxylamine (3.0 mg, 49 mol, 4.0 equiv.) was added. After stirring for 6 h, LCMS analysis revealed the presence of a product of the desired mass (LCMS: [M+H].sup.2+ m/z 837.2). Water (6.0 mL) was added and the reaction mixture was lyophilized. Purification of the crude material by semi-preparative RP-HPLC gave octasaccharide 4 as a white solid (1.7 mg, 1.01 mol, 16%). The propyl glycoside 4 had RP-HPLC (215 nm) R.sub.t=11.3 min (conditions A). .sup.1H NMR (D.sub.2O, 800 MHz) 4.81-4.79 (2d.sub.po, 3H, H-1.sub.A), 4.67-4.65 (m.sub.po, 4H, H-1.sub.A, 3H-1.sub.B), 4.54 (brd.sub.po, 2H, H-4.sub.A), 4.42-4.37 (brs, 4H, 3H-5.sub.A, H-1.sub.B), 4.31-4.30 (m.sub.po, 3H, H-5.sub.A, 2H-4.sub.A), 4.11-4.05 (m, 4H, H-3.sub.B), 4.01-3.99 (q, 4H, H-5.sub.B), 3.92-3.87 (m, 3H, H-2.sub.B, 2H-4.sub.B), 3.82-3.68 (m, 9H, 4H-2.sub.A, 3H-2.sub.B, 2H-4.sub.B), 3.67-3.65 (m.sub.po, 3H, 2H-3.sub.A, OCH.sub.2Pr), 3.59-3.55 (m.sub.po, 3H, 2H-3.sub.A, OCH.sub.2Pr), 1.96-1.89 (m.sub.po, 24H, CH.sub.3Ac), 1.47-1.44 (m, 2H, CH.sub.2Pr), 1.26 (m.sub.po, 12H, H-6.sub.B), 0.78 (t, 3H, J=7.2 Hz, CH.sub.3Pr). .sup.13C NMR (D.sub.2O, 800 MHz) 174.8, 174.6, 174.4, 173.9 (8C, CO.sub.NAc), 173.9 (4C, C-6.sub.A), 103.0 (2C, C-1.sub.A), 101.6 (2C, C-1.sub.A, C-1.sub.B), 101.1 (2C, C-1.sub.B), 100.9 (2C, C-1.sub.B, C-1.sub.A), 77.9, 77.4, 77.3 (3C, C-4.sub.A, C-4.sub.A1, C-4.sub.A2), 76.0, 75.8, 75.7, 75.5 (4C, C-3.sub.B), 72.6 (OCH.sub.2Pr), 72.0 (C-4.sub.A3), 69.1, 68.1, 67.7 (4C, C-3.sub.A), 67.4, 67.2 (4C, C-5.sub.B), 54.9, 54.7 (4C, C-4.sub.B), 51.4 (4C, C-2.sub.A), 50.9, 50.8 (4C, C-2.sub.B), 22.3, 22.1 (8C, CH.sub.3Ac), 21.2 (CH.sub.2Pr), 15.5 (4C, C-6.sub.B), 9.4 (CH.sub.3Pr). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.61H.sub.110N.sub.12O.sub.37 837.3547; found 837.3542.

    Example 4: Strategy 2.SUB.A.-NR.SUP.1.R.SUP.2.,2.SUB.B.-NDCA, 4.SUB.A.-Nap Series

    [0562] ##STR00095##

    ##STR00096##

    [0563] Allyl 4-azido-2-dichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (9). LiOH.Math.H.sub.2O (304 mg, 7.2 mmol, 3.0 equiv.) was added to the fully protected 8 (1.0 g, 2.4 mmol, 1.0 equiv.) in acetone/water (3:1, 24 mL). After stirring for 2 h at 50 C., a TLC analysis (Tol/EtOAc 1:2) indicated the total consumption of the starting material (R.sub.f 0.8) and the presence of a more polar product (R.sub.f 0.0). The reaction mixture was concentrated under reduced pressure. The crude was passed through a short silica gel column eluting with 95:5 DCM/MeOH to give the intermediate amino alcohol after extensive drying under high vacuum. The latter had HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.9H.sub.16N.sub.4O.sub.3 229.1301; found 229.1302.

    [0564] Et.sub.3N (0.5 mL, 3.6 mmol, 1.5 equiv.) was added to a solution of the crude amino alcohol in anhyd. ACN (10 mL), then cooled to 0 C. Dichloroacetyl chloride (391 L, 2.6 mmol, 1.1 equiv.) was added slowly and after stirring at this temperature for 30 min, a follow up by TLC (Tol/EtOAc 7:3) indicated the total consumption of the amino alcohol and the presence of a less polar product (R.sub.f 0.2). EtOAc (30 mL) and water (20 mL) were added and the organic layer was separated, dried over anhyd. Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The crude residue was purified by flash chromatography (Tol/EtOAc 7:3.fwdarw.6:4) to give the desired dichloroacetamide 9 (560 mg, 1.65 mmol, 68%) as a white solid. Acceptor 9 had .sup.1H NMR (DMSO-d.sub.6) 8.40 (d, 1H, J.sub.NH,2=9.2 Hz, NH), 6.38 (s, 1H, CHCl.sub.2), 5.81-5.76 (m, 1H, CH.sub.All), 5.63 (d, 1H, J=4.8 Hz, OH), 5.24-5.20 (m, 1H, CH.sub.2All), 5.10-5.07 (m, 1H, CH.sub.2All), 4.36 (d, 1H, J.sub.1,2=8.4 Hz, H-1), 4.18-4.13 (m, 1H, CH.sub.2All), 3.96-3.88 (m, 2H, H-3, CH.sub.2All), 3.76 (dd, 1H, J.sub.3,4=4.4 Hz, J.sub.4,5<1.0 Hz, H-4), 3.67-3.62 (m.sub.po, 1H, H-5, H-2), 1.20 (d, 3H, J.sub.5,6=6.4 Hz, H-6). .sup.13C NMR (DMSO-d.sub.6) 164.1 (CO.sub.NHDCA), 134.9 (CH.sub.2All), 116.7 (CH.sub.2All), 100.5 (C-1, .sup.1J.sub.C,H=161 Hz), 70.8 (C-3), 69.2 (CH.sub.2All), 68.7 (C-5), 67.6 (CHCl.sub.2), 66.2 (C-4), 53.3 (C-2), 17.7 (C-6). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.11H.sub.16Cl.sub.2N.sub.4O.sub.4Na, 361.0446; found 361.0442.

    [0565] Allyl 3-O-benzyl-2-deoxy-4-O-(2-napthylmethyl)-6-O-tert-butyldiphenylsilyl-2-tetrachlorophthalimido--L-altropyranosyl-(1.fwdarw.3)-4-azido-2-dichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (62). The crude PTFA donor 33 (737 mg, 680 mol, 1.15 equiv.) and acceptor 5 (200 mg, 592 mol, 1.0 equiv.) were mixed and co-evaporated with toluene (5 mL) twice. The mixture was dried thoroughly under high vacuum for 1 h, dissolved in anhyd. ACN (15 mL) and stirred for 30 min with freshly activated MS 4 (1.0 g) under an Ar atmosphere. After cooling to 15 C., TMSOTf (8 L, 34 L, 0.05 equiv.) was added slowly. After stirring for 1 h at this temperature, a TLC analysis (Tol/EtOAc 9:1) revealed donor consumption and the presence of a new major spot (R.sub.f 0.7). Et.sub.3N (15 L) was added and solids were filtered off. The filtrate was concentrated and the residue was purified by flash chromatography (cHex/EtOAc 90:10.fwdarw.85:15). Disaccharide 62 (490 mg, 397 mol, 67%) was obtained as white solid. The coupling product had 62 had .sup.1H NMR (CDCl.sub.3) 7.84-7.77 (m, 4H, H.sub.Ar), 7.66-7.62 (m, 4H, H.sub.Ar), 7.53-7.37 (m, 9H, H.sub.Ar), 7.04-6.97 (m, 5H, H.sub.Ar), 6.45 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.85-5.76 (m, 1H, CH.sub.All), 5.67 (s, 1H, CHCl.sub.2), 5.40 (d, 1H, J.sub.1,2=7.2 Hz, H-1.sub.A), 5.23-5.17 (m, 1H, CH.sub.2All), 5.15-5.11 (m, 1H, CH.sub.2All), 4.96 (d, 1H, J=12.6 Hz, CH.sub.2Nap), 4.82 (d.sub.po, 1H, CH.sub.2Nap), 4.82 (dd.sub.po, 1H, J.sub.2,3=11.1 Hz, H-2.sub.A), 4.62 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.59 (d, 1H, J=12.4 Hz, CH.sub.2Bn), 4.48 (dd, 1H, J.sub.3,4=3.6 Hz, J.sub.2,3=10.8 Hz, H-3.sub.B), 4.34 (pdt, 1H, J.sub.4,5=3.2 Hz, H-5.sub.A), 4.29 (dd.sub.po, 1H, J.sub.3,4=3.5 Hz, H-3.sub.A), 4.27-4.22 (m, 1H, CH.sub.2All), 4.12 (pt, H-4.sub.A), 4.08 (d, 1H, CH.sub.2Bn), 3.99-3.94 (m, 1H, CH.sub.2All), 3.84 (dd, 1H, J.sub.6a,6b=10.9 Hz, J.sub.5,6a=6.3 Hz, H-6a.sub.A), 3.80 (brd.sub.po, 1H, H-4.sub.B), 3.78 (dd.sub.po, 1H, J.sub.5,6b=6.2 Hz, H-6b.sub.A), 3.50 (ddd.sub.po, 1H, H-2.sub.B), 4.47 (dq.sub.po, 1H, J.sub.4,5=1.1 Hz, H-5.sub.B), 1.20 (d, 3H, J.sub.5,6=6.3 Hz, H-6.sub.B), 1.03 (s, 9H, CH.sub.3TBDPS). .sup.13C NMR (CDCl.sub.3) 163.9 (CO.sub.NTCA), 163.3 (CO.sub.NTCP), 139.8, 137.7, 135.7, 133.2, 133.0, 132.9, 129.5, 127.2 (C.sub.q,Ar), 133.6 (CH.sub.All), 135.6, 135.5, 129.9, 128.1, 128.0, 127.9 (2C), 127.8, 127.6, 127.5, 127.4, 126.5, 126.0, 125.9, 125.8 (C.sub.Ar), 117.7 (CH.sub.2All), 98.0 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 97.8 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 76.1 (C-5.sub.A), 75.6 (C-3.sub.B), 73.9 (C-3.sub.A), 72.7 (CH.sub.2Nap), 72.1 (CH.sub.2Bn), 71.5 (C-4.sub.A), 69.9 (CH.sub.2All), 69.1 (C-5.sub.B), 66.3 (CCl.sub.3), 65.1 (C-4.sub.B), 62.9 (C-6.sub.A), 54.6 (C-2.sub.B), 54.3 (C-2.sub.A), 26.8 (CH.sub.3TBDPS), 19.2 (C.sub.TBDPS), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.59H.sub.61Cl.sub.6N.sub.6O.sub.10Si, 1251.2350; found m/z 1251.2330.

    [0566] Allyl 2-acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-6-O-tert-butyldiphenylsilyl--L-altropyranosyl-(1.fwdarw.3)-4-azido-2-dichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (63). Ethylenediamine (75 L, 1.13 mmol, 4.0 equiv.) was added to disaccharide 62 (350 mg, 284 mol, 1.0 equiv.) in THF/MeOH (1:1, 14 mL). The reaction mixture was heated at 50 C. for 60 h at which point, a TLC analysis (Tol/EtOAc 7:3) revealed the consumption of the starting 62 (R.sub.f 0.95) and the presence of a new spot (R.sub.f 0.1). After cooling to rt, Et.sub.3N (0.5 mL) was added followed by Ac.sub.2O (268 L, 2.8 mmol, 10.0 equiv.). A TLC analysis (Tol/EtOAc 6:4) showed the presence of a new spot (R.sub.f 0.4). Solids were filtered off and washed with DCM (5 mL) twice. The filtrate was concentrated and the crude product was purified by column chromatography (cHex/EtOAc 3:1.fwdarw.2:1). Disaccharide 63, obtained as a white solid (250 mg, 247 mol, 87%), had .sup.1H NMR (CDCl.sub.3) 7.85-7.63 (m, 8H, H.sub.Ar), 7.51-7.18 (m, 15H, H.sub.Ar), 6.68 (m, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.94 (s, 1H, CHCl.sub.2), 5.91-5.82 (m, 1H, CH.sub.All), 5.29 (do, 1H, J.sub.2,NH=7.7 Hz, NH.sub.A), 5.28-5.24 (m.sub.po, 1H, CH.sub.2All), 5.20-5.16 (m, 1H, CH.sub.2All), 4.88 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.77 (d, 1H, J.sub.2,NH=12.2 Hz, CH.sub.2Nap), 7.73 (do, 1H, J.sub.1,2=3.8 Hz, H-1.sub.A), 4.72 (d.sub.po, 1H, CH.sub.2Nap), 4.71 (do, 1H, CH.sub.2Bn), 4.55 (d, 1H, J=12.2 Hz, CH.sub.2Bn), 4.47-4.39 (m, 3H, H-2.sub.A, H-3.sub.B, H-5.sub.A), 4.34-4.29 (m, 1H, CH.sub.2All), 4.07-4.02 (m, 1H, CH.sub.2All), 3.99 (dd.sub.po, 1H, J.sub.5,6a=2.7 Hz, J.sub.6a,6b=11.1 Hz, H-6a.sub.A), 3.95 (dd.sub.po, 1H, J.sub.5,6b=4.8 Hz, H-6b.sub.A), 3.91 (dd, 1H, J.sub.2,3=3.6 Hz, H-3.sub.A), 3.65 (dd, 1H, J.sub.3,4=3.6 Hz, J.sub.4,5=8.8 Hz, H-4.sub.A), 3.57 (brd, 1H, J.sub.3,4=3.4 Hz, H-4.sub.B), 3.53 (dq, 1H, J.sub.4,5=1.1 Hz, H-5.sub.B), 3.43 (ddd, 1H, J.sub.2,3=10.8 Hz, H-2.sub.B), 1.78 (s, 3H, CH.sub.3Ac), 1.18 (d, 3H, J.sub.5,6=6.3 Hz, H-6.sub.B), 1.08 (s, 9H, CH.sub.3TBDPS). .sup.13C NMR (CDCl.sub.3) 169.3 (CO.sub.NDCA), 164.7 (CO.sub.NAc), 138.8, 135.2, 133.5 (2C), 133.1, 133.0 (C.sub.q,Ar), 133.7 (CH.sub.All), 135.7, 135.6, 12.7, 129.0, 128.2 (2C), 127.8, 127.7, 127.6 (2C), 126.7, 126.1, 125.9, 125.9 (C.sub.Ar), 117.67 (CH.sub.2All), 101.5 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 97.8 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 76.5 (C-3.sub.B), 72.5 (C-3.sub.A), 70.6 (C-4.sub.A), 71.4 (CH.sub.2Nap), 70.6 (CH.sub.2Bn), 70.0 (CH.sub.2All), 69.8 (C-5.sub.A), 69.6 (C-5.sub.B), 66.5 (CHCl.sub.2), 65.6 (C-4.sub.B), 63.7 (C-6.sub.A), 55.3 (C-2.sub.B), 49.8 (C-2.sub.A), 27.0 (CH.sub.3TBDPS), 23.1 (CH.sub.3Ac), 19.4 (C.sub.TBDPS), 17.1 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.53H.sub.62Cl.sub.2N.sub.5O.sub.9Si, 1010.3694; found 1010.3669.

    [0567] Allyl 2-acetamido-3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranosyl-(1.fwdarw.3)-4-azido-2-dichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (64). TBAF (83 mg, 266 mol, 1.2 equiv.) was added to disaccharide 63 (220 mg, 221 mol, 1.0 equiv.) in anhyd. THF (10 mL) at rt. After 2 h, TLC monitoring (EtOAc), showed reaction completion and the presence of a more spot (R.sub.f 0.1). Acetic acid (27 L, 266 mol, 1.2 equiv.) was added. Volatiles were removed under reduced pressure. The residue was purified by flash chromatography (EtOAc/MeOH 100:0.fwdarw.85:15) to give alcohol 64 as a white solid (135 mg, 175 mol, 80%). Disaccharide 64 had .sup.1H NMR (DMSO-d.sub.6) 8.57 (d, 1H, J.sub.2,NH=9.2 Hz, NH.sub.B), 7.94-7.82 (m, 4H, HA, NH.sub.A), 7.76 (brs, 1H, H.sub.Ar), 7.51-7.49 (m, 2H, H.sub.Ar), 7.42-7.39 (m, 3H, H.sub.Ar), 7.32-7.25 (m, 3H, H.sub.Ar), 6.44 (s, 1H, CHCl.sub.2), 5.84-5.76 (m, 1H, CH.sub.All), 5.26-5.21 (m, 2H, CH.sub.2All), 5.12-5.09 (m, 1H, CH.sub.2All), 4.76 (brs, 1H, H-1.sub.A), 4.71 (d, 1H, J=11.6 Hz, CH.sub.2Nap), 4.66-4.61 (m, 2H, CH.sub.2Bn, OH), 4.55 (d, 1H, J=11.6 Hz, CH.sub.2Bn), 4.50 (d.sub.po, 1H, CH.sub.2Nap), 4.48 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B), 4.28-4.15 (m, 3H, H-2.sub.A, H-5.sub.A, CH.sub.2All), 4.03-3.94 (m, 3H, H-3.sub.B, H-4.sub.B, CH.sub.2All), 3.82-3.69 (m, 5H, H-2.sub.B, H-5.sub.B, H-3.sub.A, H-4.sub.A, H-6a.sub.A), 3.56-3.50 (m, 1H, H-6b.sub.A), 1.78 (s, 3H, CH.sub.3Ac), 1.23 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (DMSO-d.sub.6) 169.3 (CO.sub.NDCA), 164.2 (CO.sub.NAc), 139.3, 136.5, 133.2, 132.9 (C.sub.q,Ar), 134.9 (CH.sub.All), 128.3, 128.2, 128.1, 128.0, 127.5, 126.5, 126.4, 126.3 (2C) (C.sub.Ar), 116.8 (CH.sub.2All), 101.8 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 100.1 (C-1.sub.B, .sup.1J.sub.C,H=161 Hz), 77.7 (C-3.sub.B), 73.6 (C-3.sub.A), 72.6 (C-4.sub.A), 70.9 (CH.sub.2Nap), 70.3 (CH.sub.2Bn), 69.8 (C-5.sub.A), 69.5 (C-5.sub.B), 69.2 (CH.sub.2All), 67.6 (CHCl.sub.2), 65.4 (C-4.sub.B), 61.5 (C-6.sub.A), 52.4 (C-2.sub.B), 49.5 (C-2.sub.A), 22.9 (CH.sub.3Ac), 17.6 (C-6.sub.B). HRMS (ESI.sup.+): m/z 772.2502 (calcd for C.sub.37H.sub.43Cl.sub.2N.sub.5O.sub.9H [M+H].sup.+ m/z 772.2516).

    [0568] Allyl (benzyl 2-acetamido-3-O-benzyl-6-O-benzyl-4-O-(2-naphthylmethyl)-2-deoxy--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-dichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (65). TEMPO (4 mg, 26 mol, 0.2 equiv.), followed by BAIB (104 mg, 324 mol, 2.5 equiv.), were added to a suspension of alcohol 64 (100 mg, 130 mol, 1.0 equiv.) in DCM/water (2:1, 6.0 mL) at rt. After stirring vigorously for 2 h at rt, TLC monitoring (EtOAc/MeOH 20:1) showed the presence of a major polar spot and absence of the starting 64 (R.sub.f 0.2). 50% Aq. Na.sub.2SO.sub.3 (5 mL) was added, the DCM layer was separated, and the water phase was extracted with chloroform/isopropanol (3:1, 10 mL) twice. The water phase was acidified with dilute aq. HCl to reach pH 1 and again extracted with chloroform/isopropanol (3:1, 10 mL) twice. The combined organic phases were washed with brine (50 mL), dried by passing through a phase separator filter and concentrated under reduced pressure. The crude thus obtained was dissolved in DMF (2.0 mL) rt and benzyl bromide (44 L, 259 mol, 2.0 equiv.) followed by K.sub.2CO.sub.3 (27 mg, 194 mol, 1.5 equiv.) were added. After stirring for 4 h at rt, water (20 mL) was added. The aq. layer was washed with DCM (5.0 mL) three times. The combined DCM parts were washed with brine (20 mL), dried over Na.sub.2SO.sub.4, and concentrated under reduce pressure. The crude was purified by flash chromatography with (Tol/EtOAc 70:30.fwdarw.65:35). The benzyl ester 65, obtained as an off-white solid (70 mg, 79 mol, 61%), had R.sub.f 0.2 (Tol/EtOAc 7:3). .sup.1H NMR (CDCl.sub.3) 7.84-7.68 (m, 5H, H.sub.Ar), 7.51-7.25 (m, 12H, H.sub.Ar), 6.78 (d, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.04 (s, 1H, CHCl.sub.2), 5.91-5.81 (m, 2H, NH.sub.A, CH.sub.All), 5.37 (d, 1H, J.sub.1,2=6.0 Hz, H-1.sub.A), 5.28-5.13 (m.sub.po, 4H, CH.sub.2All, CH.sub.2Bn-6), 4.75 (d, 1H, J.sub.4,5=4.0 Hz, H-5.sub.A), 4.73 (d.sub.po, 1H, J.sub.1,2=8.8 Hz, H-1.sub.B), 4.73 (d.sub.po, 1H, CH.sub.2Nap), 4.68 (d, 1H, J=12.4 Hz, CH.sub.2Nap), 4.51 (d.sub.po, 1H, J=12.0 Hz, CH.sub.2Bn), 4.48 (d.sub.po, 1H, CH.sub.2Bn), 4.47 (dd.sub.po, J.sub.3,4=3.8 Hz, J.sub.2,3=10.9 Hz, H-3.sub.B), 4.33-4.28 (m, 1H, CH.sub.2All), 4.11-4.01 (m, 2H, H-3.sub.A, H-4.sub.A, CH.sub.2All), 3.95-3.90 (m, 2H, H-2.sub.A, H-4.sub.B), 3.52 (pdt, 1H, H-2.sub.B), 3.43 (q, 1H, H-5.sub.B), 1.88 (s, 3H, CH.sub.3Ac), 1.24 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3), 170.6 (C-6.sub.A), 169.3 (CO.sub.NDCA), 164.3 (CO.sub.NAc), 137.9, 134.9, 134.8, 133.1, 133.0 (C.sub.q,Ar), 133.6 (CH.sub.All), 133.1, 133.0 (C.sub.q,Ar), 128.7 (2C), 128.6, 128.3, 128.2, 128.1, 127.9, 127.8, 127.6, 126.8, 126.1, 126.0, 125.9 (17C, C.sub.Ar), 117.9 (CH.sub.2All), 99.3 (C-1.sub.A, .sup.1J.sub.C,H=170. Hz), 97.9 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 77.0 (C-3.sub.B), 73.4 (C-3.sub.A), 73.1 (C-4.sub.A), 72.1 (C-5.sub.A), 72.0 (2C, CH.sub.2Bn,Na.sub.p), 70.1 (CH.sub.2All), 69.2 (C-5.sub.B), 67.5 (CH.sub.2Bn-6), 66.7 (CHCl.sub.2), 65.2 (C-4.sub.B), 54.3 (C-2.sub.B), 52.5 (C-2.sub.A), 23.4 (CH.sub.3Ac), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.44H.sub.48Cl.sub.2N.sub.5O.sub.10, 876.2778; found 876.2773.

    Example 5: Strategy 2.SUB.A.-NTCA,2.SUB.B.-NTCA, TBS Series

    [0569] AB Building Block for Oligomerization: 4.sub.A-TBS

    ##STR00097##

    [0570] Allyl (benzyl 3-O-benzyl-4-O-tert-butyldimethylsilyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (6a). Imidazole (1.0 g, 14.7 mmol, 5.0 equiv.) and DMAP (50 mg, 290 nmol, 0.1 equiv.) were added to a solution of alcohol 7a (2.57 g, 2.94 mmol, 1.0 equiv.) in anhyd. THF (11.8 mL). The reaction mixture was cooled to 0 C. and tert-butyldimethylsilyl trifluoromethanesulfonate (1.69 mL, 7.35 mmol, 2.5 equiv.) was added slowly. After stirring for 2 h at rt, a TLC follow up (Tol/EtOAc 8:2) showing conversion of the starting 7a into a less polar product indicated reaction completion. MeOH (1.5 mL) was added. After stirring for 15 min at rt, volatiles were evaporated and the residue was purified by flash chromatography (cHex/EtOAc 90:10.fwdarw.80:20) to give the fully protected 6a (2.62 g, 2.65 mmol, 90%). Disaccharide 6a had R.sub.f 0.3 (Tol/EtOAc 8:2). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.39H.sub.49Cl.sub.6N.sub.5O.sub.10SiNa, 1008.1277; found 1008.1379.

    [0571] (Benzyl 3-O-benzyl-4-O-tert-butyldimethylsilyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranose (8a). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (27 mg, 30 nmol, 0.02 equiv.) was dissolved in anhyd. THF (3.2 mL) and stirred for 20 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly with Ar and transferred by means of a cannula into a solution of allyl glycoside 6a (1.58 g, 1.6 mmol, 1.0 equiv.) in anhyd. THF (8.0 mL). The reaction mixture was stirred for 2 h at rt, at which time a solution of I.sub.2 (811 mg, 3.2 mmol, 2.0 equiv.) in THF/H.sub.2O (4:1, 7.8 mL) was added. After stirring for 45 min at rt, a TLC analysis (Tol/EtOAc 95:5) revealed the full consumption of the isomerization product (R.sub.f 0.4) and the presence of two more polar spots (R.sub.f 0.25, 0.1). 10% Aq. Na.sub.2SO.sub.3 (50 mL, 20 equiv.) was added and volatiles were evaporated. The aq. phase was extracted with DCM (200 mL) twice. The combined organic layers were washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. Purification of the residue by flash chromatography (cHex/EtOAc 80:20.fwdarw.40:60) gave the expected hemiacetal 8a (1.42 g, 1.5 mmol, 95%) as a white floppy solid. Hemiacetal 8a (/ 5:1) had R.sub.f 0.3, 0.25 (cHex/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.36H.sub.45Cl.sub.6N.sub.5O.sub.10SiNa, 968.0964; found 968.0972.

    [0572] (Benzyl 3-O-benzyl-4-O-tert-butyldimethylsilyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl (N-phenyl)trifluoroacetimidate (9a) and 2-trichloromethyl [(benzyl 3-O-benzyl-2-deoxy-4-O-tert-butyldimethylsilyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-1,2,4,6-tetradeoxy--D-galactopyrano]-[2,1,d]-oxazoline (10a). PTFACl (970 L, 6.1 mmol, 2.0 equiv.) and Cs.sub.2CO.sub.3 (1.19 g, 3.67 mmol, 1.2 equiv.) were added to hemiacetal 8a (2.9 g, 3.06 mmol, 1.0 equiv.) in acetone (34 mL). After stirring for 1 h at rt, the reaction mixture was filtered through a pad of Celite and washed with DCM (50 mL) twice. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography (cHex/Et.sub.3N 99:1 for column equilibration, then cHex/EtOAc 100:0.fwdarw.90:10) to give the expected donor as a 6:4 mix of PTFA 9a and oxazoline 10a (3.09 g, 91%). The PTFA donor 9a had R.sub.f 0.4 (cHex/EtOAc 85:15).

    [0573] Oxazoline 10a had R.sub.f 0.45 (cHex/EtOAc 85:15). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.36H.sub.44Cl.sub.6N.sub.5O.sub.9Si, 928.1039; found 928.1004.

    [0574] Oliomerization from Building Block 6a (4.sub.A-TBS)

    ##STR00098##

    [0575] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-tert-butyldimethylsilyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (11a). Freshly activated MS 4 (2.63 g) was added to a mix of donors 9a and 10a (3.08 g, 2.75 mmol, 1.1 equiv. theo.) and disaccharide acceptor 7a (2.19 g, 2.51 mmol, 1.0 equiv.) in anhyd. DCE (52 mL) and the suspension was stirred for 30 min under an Ar atmosphere at rt. After cooling to 30 C., TMSOTf (45 L, 250 mol, 0.1 equiv.) was added slowly and stirring went on for 40 min at this temperature. A TLC analysis (Tol/EtOAc 8:2) showed some remaining acceptor 7a in minor amount and the presence of a new spot (R.sub.f 0.5). Et.sub.3N (10 L, 1.0 equiv.) was added. The suspension was filtered through a fitted funnel and washed with DCM (50 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/EtOAc 95:5.fwdarw.90:10) to give tetrasaccharide 11a as a white solid (3.48 g, 1.93 mmol, 77%). The coupling product 11a had R.sub.f 0.35 (Tol/EtOAc 9:1). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.69H.sub.78Cl.sub.12N.sub.10O.sub.19SiNa, 1821.1375; found 1821.1779.

    [0576] (Benzyl 3-O-benzyl-2-deoxy-4-O-tert-butyldimethylsilyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranose (12a). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (9.4 mg, 11 nmol, 0.02 equiv.) was dissolved in anhyd. THF (1.1 mL) and stirred for 20 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly with Ar and transferred by means of a cannula into a solution of allyl glycoside 11a (1.0 g, 554 nmol, 1.0 equiv.) in anhyd. THF (2.8 mL). The reaction mixture was stirred for 2 h at rt, at which time a solution of I.sub.2 (281 mg, 1.11 mmol, 2.0 equiv.) in THF/H.sub.2O (4:1, 3.3 mL) was added. After stirring for 1 h at rt, a TLC analysis (Tol/EtOAc 8:2) revealed the full consumption of the isomerization product (R.sub.f 0.5) and the presence of two more polar spots (R.sub.f 0.3, 0.1). 10% Aq. Na.sub.2SO.sub.3 (50 mL, 20 equiv.) was added and volatiles were evaporated. The aq. phase was extracted with DCM (60 mL) thrice. The combined organic layers were washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. Purification of the residue by flash chromatography (cHex/EtOAc 80:20.fwdarw.60:40) gave the expected hemiacetal 12a (916 mg, 94%) as a white floppy solid. Hemiacetal 12a (/ 5:1) had R.sub.f 0.3, 0.1 (Tol/EtOAc 8:2). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.66H.sub.74Cl.sub.12N.sub.10O.sub.19SiNa, 1781.1061; found 1781.1320.

    [0577] (Benzyl 3-O-benzyl-2-deoxy-4-O-tert-butyldimethylsilyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl (N-phenyl)trifluoroacetimidate (13a). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (32 mg, 38 nmol, 0.04 equiv.) was dissolved in anhyd. THF (3.9 mL) and stirred for 20 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly with Ar and transferred by means of a cannula into a solution of allyl glycoside 11a (1.72 g, 953 nmol, 1.0 equiv.) in anhyd. THF (4.5 mL). The reaction mixture was stirred for 2 h at rt, at which time a solution of 12 (484 mg, 1.9 mmol, 2.0 equiv.) in THF/H.sub.2O (4:1, 5.7 mL) was added. After stirring for 1.5 h at rt, a TLC analysis (Tol/EtOAc 8:2) revealed the full consumption of the isomerization product (R.sub.f 0.5) and the presence of two more polar spots (R.sub.f 0.3, 0.1). 10% Aq. Na.sub.2SO.sub.3 (50 mL, 20 equiv.) was added and volatiles were evaporated. The aq. phase was extracted with DCM (200 mL) twice. The combined organic layers were washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. Flash chromatography (cHex/EtOAc 80:20.fwdarw.40:60) f the residue gave the expected hemiacetal 12a (1.66 g, 99%) as a white floppy solid.

    [0578] PTFACl (287 L, 1.81 mmol, 2.0 equiv.) and Cs.sub.2CO.sub.3 (355 mg, 1.09 mmol, 1.2 equiv.) were added to hemiacetal 12a (1.6 g, 910 mol, 1.0 equiv.) in acetone (10 mL). After stirring for 1.5 h at rt, a TLC (cHex/EtOAc 7:3) follow up revealed that conversion was complete. The reaction mixture was filtered through a pad of Celite and washed with DCM (50 mL) twice. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography (cHex/EtOAc 100:0.fwdarw.70:30) to give the expected donor as a mix of PTFA 13a and oxazoline 14a (1.43 g, 84% over two steps). The PTFA donor 13a had R.sub.f 0.55 (cHex/EtOAc 7:3).

    [0579] Oxazoline 14a had R.sub.f 0.65 (cHex/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.66H.sub.76Cl.sub.12N.sub.11O.sub.18Si, 1758.1401; found 1758.1414.

    [0580] Allyl (benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (15a). Et.sub.3N.Math.3HF (1.25 mL, 7.67 mmol, 8.0 equiv.) was added to a solution of the fully protected tetrasaccharide 11a (1.73 g, 958 mol, 1.0 equiv.) in anhyd. THF (3.45 mL), and the solution was stirred at rt for 48 h at which time a TLC follow up (Tol/EtOAc 8:2) showed reaction completion. MeOH (1.0 mL) was added and volatiles were eliminated under vacuum. Flash chromatography (Tol/EtOAc 80:20.fwdarw.75:25, then Tol/Acet 50:50.fwdarw.20:80) of the residue gave the desired alcohol 15a (1.55 g, 96%). The tetrasaccharide acceptor 15a had R.sub.f 0.45 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+H+K].sup.2+ calcd for C.sub.63H.sub.63Cl.sub.12N.sub.10O.sub.19K, 862.0164; found 862.0118.

    [0581] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-tert-butyldimethylsilyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (16a). Freshly activated MS 4 (332 mg) was added to a mix of donors 9a and 10a (414 mg, 370 mol, 1.25 equiv. theo.) and tetrasaccharide acceptor 15a (500 mg, 296 mol, 1.0 equiv.) in anhyd. DCE (6.7 mL) and the suspension was stirred for 30 min under an Ar atmosphere at rt. After cooling to 0 C. and stirring for 10 min, TMSOTf (6.6 L, 30 mol, 0.1 equiv., 20 L from a TMSOTf/DCE solution (1:2.7 v/v)) was added and stirring went on for 45 min at 0 C. A TLC analysis (Tol/EtOAc 8:2) showed some remaining acceptor 15a in minor amount (R.sub.f 0.15) and the presence of a new spot (R.sub.f 0.35). After stirring for an additional 15 min, Et.sub.3N (50 L of a Et.sub.3N/DCE solution (1:2 v/v)) was added. The suspension was filtered through a fitted funnel and washed with DCM (50 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/EtOAc 90:10.fwdarw.70:30) to give hexasaccharide 16a as a white solid (599 mg, 223 mol, 77%). HRMS (MALDI): m/z [M+Na].sup.+ calcd for C.sub.99H.sub.107C18N.sub.15O.sub.28SiNa, 2635.1499; found 2635.0943. HRMS (MALDI): m/z [M+K].sup.+ calcd for C.sub.99H.sub.107Cl.sub.18N.sub.15O.sub.28SiK 2652.1191; found 2652.0420.

    [0582] Allyl (benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (17a). Et.sub.3N.Math.3HF (103 L, 635 mol, 8.0 equiv.) was added to a solution of the fully protected hexasaccharide 16a (208 mg, 79 mol, 1.0 equiv.) in anhyd. THF (290 L), and the solution was stirred at rt for 48 h at which time a TLC follow up (Tol/EtOAc 8:2) showed reaction completion. MeOH (1.0 mL) was added and volatiles were eliminated under vacuum. Flash chromatography (Tol/EtOAc 80:20.fwdarw.50:50) of the residue gave the desired alcohol 17a (176 mg, 89%). The hexasaccharide acceptor 17a had R.sub.f 0.35 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.93H.sub.97Cl.sub.18N.sub.16O.sub.28 2517.1033; found 2516.7451. HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.93H.sub.97Cl.sub.18N.sub.16O.sub.28 1267.5687; found 1267.5885.

    [0583] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-tert-butyldimethylsilyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (18a). [2+6 glycosylation] Freshly activated MS 4 (13 mg) was added to a mix of donors 9a and 10a (17 mg, 15 mol, 1.25 equiv.) and hexasaccharide acceptor 17a (30 mg, 12 mol, 1.0 equiv.) in anhyd. DCE (2.5 mL) and the suspension was stirred for 30 min under an Ar atmosphere at rt. After cooling to 0 C. and stirring for 15 min, TMSOTf (0.2 L, 1.2 mol, 0.1 equiv., 10 L of a TMSOTf/DCE solution (1:49 v/v)) was added and stirring went on for 40 min at 10 C. More TMSOTf (0.1 equiv.) was added. After stirring for 20 min at 10 C., a TLC analysis (Tol/EtOAc 8:2) some remaining acceptor 17a and a new spot (R.sub.f 0.35), albeit no further evolution. Et.sub.3N (0.1 equiv., 10 L, of a Et.sub.3N/DCE solution (1:49 v/v)) was added. The suspension was filtered through a fitted funnel and washed with DCM (20 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/EtOAc 90:10.fwdarw.70:30) to give octasaccharide 18a as a white solid (16.5 mg, 47 mol, 40%) and some remaining hexasaccharide acceptor 17a (12 mg, 40%).

    [0584] [4+4 glycosylation] Freshly activated MS 4 (124 mg) was added to a mix of donors 13a and 14a (126 mg, 65 mol, 1.1 equiv.) and tetrasaccharide acceptor 15a (100 mg, 59 mol, 1.0 equiv.) in anhyd. DCE (2.5 mL) and the suspension was stirred for 45 min under an Ar atmosphere at rt. After cooling to 10 C. and stirring for 15 min, TMSOTf (0.3 L, 2 mol, 0.03 equiv., 15 L of a TMSOTf/DCE solution (1:49 v/v)) was added and stirring went on for 20 min at 10 C. A TLC analysis (Tol/EtOAc 8:2) showed the presence of acceptor 15a in minor amount and a new spot (R.sub.f 0.35). After stirring for an additional 15 min, Et.sub.3N (20 L of a Et.sub.3N/DCE solution (1:36.5 v/v)) was added. The suspension was filtered through a fitted funnel and washed with DCM (50 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/EtOAc 95:5.fwdarw.70:30) to give octasaccharide 18a as a white solid (163 mg, 47 mol, 81%). The coupling product 18a had R.sub.f 0.5 (Tol/EtOAc 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.129H.sub.142Cl.sub.24N.sub.22O.sub.37Si, 1732.6169; found 1732.6240.

    [0585] Allyl (benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (19a). Et.sub.3N.Math.3HF (75 L, 455 mol, 5.0 equiv.) was added to a solution of the fully protected octasaccharide 18a (312 mg, 91 mol, 1.0 equiv.) in anhyd. THF (455 L), and the solution was stirred at rt for 52 h at which time a TLC follow up (Tol/EtOAc 8:2) showed that only minor traces of the starting 18a and the presence of a major more polar product. MeOH (300 L) was added and after 10 min at rt, volatiles were eliminated under vacuum. The residue was taken in DCM (100 mL) and the organic phase was washed with satd aq. NaHCO.sub.3 (30 mL) and brine (30 mL), dried on Na.sub.2SO.sub.4, and filtered. After concentration under vacuum, flash chromatography (Tol/EtOAc 80:20.fwdarw.50:50) of the residue gave the desired alcohol 19a (269 mg, 81 mol, 89%). The octasaccharide acceptor 19a had R.sub.f 0.3 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.123H.sub.128Cl.sub.24N.sub.22O.sub.37Si, 1675.5724; found 1676.5715.

    [0586] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-tert-butyldimethylsilyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (20a). Freshly activated MS 4 (227 mg) was added to a mix of donors 13a and 14a (139 mg, 72 mol, 1.2 equiv.) and hexasaccharide acceptor 17a (150 mg, 60 mol, 1.0 equiv.) in anhyd. DCE (4.5 mL) and the suspension was stirred for 1 h under an Ar atmosphere at rt. After cooling to 10 C. and stirring for 15 min, TMSOTf (0.3 L, 2 mol, 0.03 equiv., 15 L of a TfOH/DCE solution (1:49 v/v)) was added and stirring went on for 20 min at 10 C. A TLC analysis (Tol/EtOAc 8:2) showed the presence of a new spot (R.sub.f 0.35) and stirring went on for another 20 min at 10 C. Et.sub.3N (30 L of a Et.sub.3N/DCE solution (1:42 v/v)) was added. Solids were filtered and washed with DCM (20 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/EtOAc 95:5.fwdarw.70:30) to give decasaccharide 20a as a white solid (206 mg, 48 mol, 81%) in addition to contaminated fractions. The fully protected decasaccharide 20a had R.sub.f 0.5 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.159H.sub.171Cl.sub.30N.sub.27O.sub.46Si, 1675.5724; found 1676.5715.

    [0587] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-tert-butyldimethylsilyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (21a). Route 1. Freshly activated MS 4 (137 mg) was added to a mix of donors 13a and 14a (50 mg, 26 mol, 1.3 equiv.) and octasaccharide acceptor 19a (66 mg, 20 mol, 1.0 equiv.) in anhyd. DCE (2.7 mL) and the suspension was stirred for 30 min under an Ar atmosphere at rt. After cooling to 0 C. and stirring for 10 min, TMSOTf (0.14 L, 1 mol, 0.03 equiv., 10 L of a TfOH/DCE solution (1:70 v/v)) was added and stirring went on for 1 h at 0 C. A TLC analysis (Tol/EtOAc 8:2) showed the presence of a new spot (R.sub.f 0.35). Et.sub.3N (80 L of a solution in DCE (1:35 v/v)) was added. Solids were filtered and washed with DCM (20 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/EtOAc 90:10.fwdarw.70:30) to give dodecasaccharide 21a as a white solid (51 mg, 10 mol, 50%) in addition to contaminated fractions and a less polar compound corresponding to the silylated acceptor (9 mg, 2.6 mol, 13%). The silylated side-product had R.sub.f 0.65 (Tol/EtOAc 75:25). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.126H.sub.138C.sub.124N.sub.22O.sub.37Si, 1712.0916; found 1712.0918.

    [0588] Route 2. Freshly activated MS 4 (310 mg) was added to a mix of donors 13a and 14a (57 mg, 29 mol, 1.3 equiv.) and octasaccharide acceptor 19a (75 mg, 23 mol, 1.0 equiv.) in anhyd. DCE (3.1 mL) and the suspension was stirred for 1 h under an Ar atmosphere at rt. After cooling to 10 C. and stirring for 5 min, TfOH (0.1 L, 1 mol, 0.03 equiv., 10 L of a TfOH/DCE solution (1:125 v/v)) was added and stirring went on for 25 min at 10 C. A TLC analysis (Tol/EtOAc 8:2) showed the presence of acceptor 19a in minor amount and a new spot (R.sub.f 0.35). After stirring for an additional 35 min at 10 C., Et.sub.3N (40 L of a Et.sub.3N/DCE solution (1:50 v/v)) was added. The suspension was filtered through a fitted funnel and washed with DCM (50 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/EtOAc 90:10.fwdarw.70:30) to give dodecasaccharide 21a as a white solid (66 mg, 13 mol, 56%) in addition to contaminated fractions. The coupling product 21a had R.sub.f 0.3 (Tol/EtOAc 75:25). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C189H.sub.200Cl.sub.36N.sub.32O.sub.55Si, 2550.1213; found 2550.0933.

    Example 6: Strategy 2.SUB.A.-NTCA,2.SUB.B.-NTCA, 4.SUB.A.-Nap

    [0589] The Ready-for-Oligomerization 4.sub.A-Nap AB Building Block

    ##STR00099## ##STR00100##

    [0590] Allyl 2-amino-3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-(2-naphthylmethyl)--L-altropyranoside (1a). A solution of the azido precursor 30 (1.0 g, 1.4 mmol) in THF (7.0 mL) was treated with PPh.sub.3 (0.4 g, 1.5 mmol, 1.1 equiv.) and H.sub.2O (0.5 mL, 28 mmol, 20.0 equiv.). The reaction mixture was heated to 60 C. and stirred overnight. Follow up by TLC (Tol/EtOAc 9:1) indicated the total conversion of the starting material (R.sub.f 0.7) and the presence of a more polar product (R.sub.f 0.1). The reaction mixture was concentrated and coevaporated repeatedly with toluene under reduced pressure. The crude material was purified by flash chromatography (Tol/EtOAc/NH.sub.3OH 85:15:2) to give amine 1a (0.73 g, 73%) as a colorless oil. .sup.1H NMR (CDCl.sub.3) 7.88-7.17 (m, 22H, H.sub.Ar), 6.02-5.91 (m, 1H, CHCH.sub.2), 5.31 (dq, 1H, CHCH.sub.2), 5.19 (dq, 1H, CHCH.sub.2), 4.79 (d, 2H, CH.sub.2Nap), 4.58 (d, 1H, J.sub.1,2=5.1 Hz, H-1), 4.54 (dd, 2H, CH.sub.2Bn), 4.34 (dd, 1H, J.sub.5,6=5.5 Hz, J.sub.5,4=10.4 Hz, H-5), 4.33 (ddt, 1H, CH.sub.2All), 4.05 (ddt, 1H, CH.sub.2All), 4.04-4.01 (m, 1H, H-4), 3.89 (dd, 1H, J.sub.6b,5=4.9 Hz, J.sub.6b,6a=10.6 Hz, H-6b), 3.84 (dd, 1H, J.sub.6a,5=5.0 Hz, J.sub.6a,6b=10.6 Hz, H-6a), 3.66 (dd, 1H, J.sub.3,4=3.3 Hz, J.sub.3,2=8.2 Hz, H-3), 3.45 (dd, 1H, J.sub.2,1=5.2 Hz, J.sub.2,3=8.0 Hz, H-2), 1.08 (s, 9H, H-.sup.tBu.sub.TBDPS). .sup.13C NMR (CDCl.sub.3) 135.9 (C.sub.q,Nap), 135.7 (C.sub.qPh,TBDPS), 135.6 (C.sub.qPh,TBDPS), 134.9 (C.sub.qBn), 134.4 (CHCH.sub.2), 133.9 (C.sub.q,Nap), 133.7 (C.sub.q,Nap), 129.8-125.3 (C.sub.Ar,Bn,Ph,Nap), 117.1 (CHCH.sub.2), 101.4 (C-1.sub.A, .sup.1J.sub.C,H=163.7 Hz), 78.0 (C-3), 73.0 (C-5), 71.7 (CH.sub.2Nap), 71.6 (CH.sub.2Bn), 71.0 (C-4), 68.9 (CH.sub.2All), 63.3 (C-6), 51.9 (C-2), 26.8 (CH.sub.3tBu, TBDPS), 19.3 (C.sub.qtBu,TBDPS).

    [0591] Allyl 3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranoside (1b). Zn (24.7 g, 378 mmol, 8.0 equiv.) and AcOH (21.6 mL, 378 mmol, 10 equiv.) was added to the azido precursor 30 in anhyd. THF (470 mL). After stirring for 1 h, a TLC analysis (Tol/EtOAc 10:1) showed the absence of azide 1 (R.sub.f 0.85) and the presence of a more polar spot. The suspension was filtered over a pad of Celite and washed with DCM. The DCM layer was washed with satd aq. NaHCO.sub.3, water, and brine, dried over Na.sub.2SO.sub.4, concentrated under reduced pressure, and dried under high vacuum.

    [0592] The crude amine 1a was dissolved in DCM (100 mL) and cooled to 0 C. Et.sub.3N (9.8 mL, 70.8 mmol, 1.5 equiv.) was added followed by the dropwise addition of trichloroacetyl chloride (6.85 mL, 61.4 mmol, 1.3 equiv.) while maintaining the temperature at 0 C. After stirring for 1 h, a TLC follow up (Tol/EtOAc) indicated reaction completion. The reaction was quenched by addition of methanol (2.0 mL). Following dilution with DCM (200 mL), washing with 50% brine, the DCM layer was separated, dried and concentrated. Flash chromatography of the crude (cHex/EtOAc 54:6) gave trichloroacetamide 1b as a colorless oil (31.6 g, 38.0 mmol, 80%). Allyl glycoside 1b had R.sub.f 0.2 (Tol/EtOAc 7:3). .sup.1H NMR (CDCl.sub.3) 7.85-7.83 (m, 1H, H.sub.Ar), 7.77-7.72 (m, 6H, H.sub.Ar), 7.63 (brs, 1H, H.sub.Ar), 7.52-7.28 (m, 14H, H.sub.Ar), 6.75 (d, 1H, J.sub.2,NH=8.4 Hz, NH), 5.97-5.89 (m, 1H, CH.sub.All), 5.37-5.32 (m, 1H, CH.sub.2All), 5.24-5.20 (m, 1H, CH.sub.2All), 4.92 (d, 1H, J=12.0 Hz, CH.sub.2Nap), 4.88 (d.sub.po, 1H, J.sub.1,2=4.8 Hz, H-1), 4.73 (d, 2H, J=12.0 Hz, CH.sub.2Bn), 4.72 (d.sub.po, 1H, CH.sub.2Nap), 4.61 (d, 1H, CH.sub.2Bn), 4.51-4.48 (m, 1H, CH.sub.2All), 4.51 (ddd, 1H, J.sub.2,3=1.0 Hz, H-2), 4.37 (dt, J.sub.5,6b=2.8 Hz, H-5), 4.29-4.24 (m, 1H, CH.sub.2All), 4.14 (dd, 1H, J.sub.5,6a=3.2 Hz, J.sub.6a,6b=11.2 Hz, H-6a), 4.06-3.97 (m, 3H, H-3, H-6b, CH.sub.2All), 3.93 (dd, J.sub.3,4=3.2 Hz, J.sub.4,5=9.2 Hz, H-4), 1.00 (s, 9H, CH.sub.3, TBDPS). .sup.13C NMR (CDCl.sub.3) 161.2 (CO.sub.NHTCA), 137.9, 133.2 (2C), 133.0, 129.0, 125.3 (C.sub.q,Ar), 133.6 (CH.sub.All), 135.8, 135.5, 129.7, 128.3, 128.2, 127.9, 127.7 (2C), 127.6, 126.7, 126.1, 125.9, 125.8 (22C, C.sub.Ar), 117.3 (CH.sub.2All), 97.3 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 92.1 (CCl.sub.3), 72.0 (C-3), 71.5 (CH.sub.2Nap), 70.9 (CH.sub.2Bn), 70.5 (C-4), 68.6 (C-5), 68.3 (CH.sub.2All), 62.8 (C-6), 50.8 (C-2), 26.9 (CH.sub.3, TBDPS), 19.4 (C.sub.TBDPS). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.45H.sub.52Cl.sub.3N.sub.2O.sub.6Si, 849.2655; found 849.2657.

    [0593] 3-O-Benzyl-2-deoxy-4-O-(2-napthylmethyl)-6-O-tert-butyldiphenylsilyl-2-trichloroacetamido-/-L-altropyranose (2b). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (965 mg, 1.14 mmol, 0.03 equiv.) was dissolved in anhyd. THF (100 mL) and stirred for 30 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly with Ar and transferred by means of a cannula into a solution of allyl glycoside 1b (31.6 g, 38.0 mmol, 1.0 equiv.) in anhyd. THF (200 mL). The reaction mixture was stirred for 4 h at rt, at which time a solution of NIS (11.2 g, 49.4 mmol, 1.3 equiv.) in H.sub.2O (60 mL) was added. After stirring for 1 h at rt, a TLC analysis (Tol/EtOAc 9:1) revealed the full consumption of the isomerization product (R.sub.f 0.7) and the presence of a more polar spot (R.sub.f 0.4). 10% Aq. Na.sub.2SO.sub.3 was added and volatiles were evaporated. The aq. phase was extracted with DCM (200 mL) twice. The combined organic layers were washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. Purification of the residue by flash chromatography (cHex/EtOAc 90:10.fwdarw.88:12) yielded the expected hemiacetal 2b (24.6 g, 31.0 mmol, 82%) as a white floppy solid. Hemiacetal 2b (/ 5:1) had R.sub.f 0.5 (cHex/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.85-7.64 (m, 10.4H, H.sub.Ar), 7.52-7.28 (m, 18.6H, H.sub.Ar), 7.04 (d, 1H, J.sub.2,NH=5.6 Hz, NH.sub.), 6.80 (d, 0.24H, J.sub.2,NH=5.6 Hz, NH.sub.), 5.64 (d, 0.25H, J.sub.1,OH=11.2 Hz, OH.sub.), 5.41 (dd.sub.po, 1H, J.sub.1,2=3.6 Hz, H-1.sub.), 5.10 (d, 0.25H, J.sub.1,2=1.0 Hz, H-1.sub.), 4.90 (d, 0.29H, J=11.2 Hz, CH.sub.2Nap,), 4.82-4.74 (m, 2.35H, CH.sub.2Nap,, CH.sub.2Nap,), 4.69 (d.sub.po, 1.5H, CH.sub.2Bn,, CH.sub.2Bn,), 4.61 (d, 1H, J=11.6 Hz, CH.sub.2n,), 4.39 (ddd, 0.29H, J.sub.2,3=3.6 Hz, H-2), 4.30-4.24 (brm, 2.56H, H-2.sub., H-3.sub., H-5.sub., H-5), 4.20 (dd, 0.27H, J.sub.5,6b=2.0 hz, J.sub.6a,6b=11.2 Hz, H-6a.sub.), 4.11-3.90 (m, 4.71H, H-3.sub., H-4.sub., H-4.sub., H-6a.sub., HH-6b.sub., H-6b.sub.), 2.98 (d, 1H, J.sub.1,OH=4.0 Hz, OH.sub.), 1.12 (s, 2.22H, CH.sub.3, TBDPS), 1.09 (s, 9H, CH.sub.3, TBDPS). .sup.13C NMR (CDCl.sub.3) 162.1 (CO.sub.NHTCA,), 161.4 (CO.sub.NHTCA,), 137.8, 136.5, 134.9, 134.8, 133.7, 133.4, 133.1 (2C), 133.0, 132.9 (C.sub.q,Ar), 135.8, 135.7, 135.5 (2C), 129.7 (3C), 128.7, 128.4, 128.3, 128.2, 128.0, 127.9 (2C), 127.7 (3C), 127.6 (2C), 126.6, 126.2, 126.1, 125.9, 125.7, 125.6 (C.sub.Ar), 93.4 (C-1.sub., .sup.1J.sub.C,H=175 Hz), 91.3 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 92.2, 91.8 (CCl.sub.3), 74.3 (CH.sub.2Nap,), 74.2 (C-3.sub., C-3), 73.9 (C-5.sub.), 72.9 (CH.sub.2Nap,), 72.1 (CH.sub.2Bn,), 72.1 (C-5.sub.), 71.7 (CH.sub.2n,), 71.3 (C-4.sub.), 70.8 (C-4.sub.), 67.7 (C-5.sub.), 63.0 (C-6.sub.), 62.5 (C-6.sub.), 53.1 (C-2.sub.), 51.3 (C-2.sub.), 27.0, 26.9 (C.sub.TBDPS), 19.4 (CH.sub.3,TBDPS). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.44H.sub.46Cl.sub.3N.sub.2O.sub.6Si, 809.2342; found 809.2346.

    [0594] 3-O-Benzyl-2-deoxy-4-O-(2-napthylmethyl)-6-O-tert-butyldiphenylsilyl-2-trichloroacetamido-/-L-altropyranosyl N-(phenyl)trifluoroacetimidate (3b). PTFACl (7.74 mL, 37.3 mmol, 1.2 equiv.) and Cs.sub.2CO.sub.3 (11.1 g, 34.2 mmol, 1.1 equiv.) were added to hemiacetal 2b (24.6 g, 31.0 mmol, 1.0 equiv.) in acetone (300 mL). After stirring for 2 h at rt, the reaction mixture was filtered through a pad of Celite and washed with DCM (50 mL) twice. The filtrate was concentrated under reduced pressure and dried under vacuum to give the crude donor 3b (30.0 g, 31.0 mmol, quant.), which was used as such in the next step. The PTFA donor 3b had R.sub.f 0.92 (Tol/EtOAc 9:1). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.50H.sub.48Cl.sub.3F.sub.3N.sub.2O.sub.6SiNa, 985.2197; found 985.2191.

    [0595] Allyl (3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyl)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (4b). A mix of the crude PTFA donor 3b (30.0 g, 31.0 mmol, 1.0 equiv. theo.) and acceptor 8 (10.8 g, 32.0 mmol, 1.03 equiv.) were co-evaporated with anhyd. toluene (50 mL) and then dried under high vacuum. Freshly activated MS 4 (15.0 g) was added to a solution of the starting materials in anhyd. DCM (520 mL) and the suspension was stirred for 1 h under an Ar atmosphere at rt. After cooling to 15 C., TMSOTf (281 L, 0.05 equiv.) was added slowly and stirring went on for 40 min during which the bath temperature was kept at 15 C. A TLC analysis (Tol/EtOAc 9:1) showed the absence of donor 3b and the presence of a new spot (R.sub.f 0.6) in addition to a slight amount of acceptor 8 (R.sub.f 0.05). At completion, Et.sub.3N (300 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (100 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (cHex/EtOAc 90:10.fwdarw.85:15) to give disaccharide 4b as a white solid (35.6 g, 31.0 mmol, 99%). The coupling product 4b had .sup.1H NMR (CDCl.sub.3) 7.82-7.80 (m, 1H, H.sub.Ar), 7.74-7.68 (m, 6H, H.sub.Ar), 7.61 (brs, 1H, H.sub.Ar), 7.54-7.28 (m, 14H, H.sub.Ar), 6.75 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B), 6.49 (d, 1H, J.sub.2,NH=8.4 Hz, NH.sub.A), 5.91-5.81 (m, 1H, CH.sub.All), 5.30-5.25 (m, 1H, CH.sub.2All), 5.21-5.18 (m, 1H, CH.sub.2All), 4.92 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.83-4.75 (m, 3H, H-1.sub.A, CH.sub.2Nap), 4.73 (d, 1H, J=12.4 Hz, CH.sub.2Bn), 4.60 (d, 1H, CH.sub.2Bn), 4.55-4.48 (m.sub.po, 2H, H-3.sub.B, H-5.sub.A), 4.37-4.31 (m, 2H, H-2.sub.A, CH.sub.2All), 4.09-3.97 (m, 3H, H-6ab.sub.A, CH.sub.2All), 3.92 (t, 1H, J.sub.2,3=J.sub.3,4=3.2 Hz, H-3.sub.A), 3.78 (dd, 1H, J.sub.4,5=9.6 Hz, H-4.sub.A), 3.58-3.54 (m, 2H, H-5.sub.B, H-4.sub.B), 3.45 (ddd.sub.po, 1H, H-2.sub.B), 1.21 (d, 3H, 6.4 Hz, H-6.sub.B), 1.10 (s, 9H, CH.sub.3,TBDPS). .sup.13C NMR (CDCl.sub.3) 162.2, 161.0 (CO.sub.NHTCA), 138.3, 134.8, 133.5, 133.3, 133.1 (C.sub.q,Ar), 133.5 (CH.sub.All), 135.8, 135.5, 129.8, 128.8, 128.3 (2C), 127.8, 127.7 (2C), 127.6, 126.9, 126.2, 126.0, 125.8 (22C, C.sub.Ar), 117.8 (CH.sub.2All), 100.5 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 97.3 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 92.1, 91.8 (2C, CCl.sub.3), 76.2 (C-3.sub.B), 72.6 (C-3.sub.A), 72.1 (CH.sub.2Nap), 70.8 (CH.sub.2Bn), 70.1 (CH.sub.2All), 70.1 (C-4.sub.A), 69.7 (C-5.sub.B), 69.4 (C-5.sub.A), 65.5 (C-4.sub.B), 63.2 (C-6.sub.A), 55.9 (C-2.sub.B), 51.2 (C-2.sub.A), 27.0 (CH.sub.3,TBDPS), 19.5 (C.sub.TBDPS), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.53H.sub.61Cl.sub.6N.sub.6O.sub.9Si, 1163.2395; found 1163.2402.

    [0596] Allyl (3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyl)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (5b). TBAF.Math.3H.sub.2O (10.7 g, 34.1 mmol, 1.1 equiv.) was added to disaccharide 4b (35.6 g, 31.0 mmol, 1.0 equiv.) in THF (300 mL) and the reaction mixture was stirred at rt for 4 h. A TLC analysis (Tol/EtOAc 1:4) showed the consumption of the fully protected 4b (R.sub.f 1.0) and the presence of a polar spot. Acetic acid (2.1 mL, 3.73 mmol, 1.2 equiv.) was added and after stirring for 10 min, volatiles were evaporated. The residue was purified by flash chromatography (cHex/EtOAc 40:60.fwdarw.0:100) to give alcohol 5b (25.2 g, 27.7 mmol, 89%) as a white solid. Disaccharide 5b had R.sub.f 0.25 (Tol/EtOAc 1:1). .sup.1H NMR (CDCl.sub.3) 7.83-7.73 (m, 3H, H.sub.Ar), 7.60 (brs, 1H, H.sub.Ar), 7.53-7.46 (m, 4H, H.sub.Ar), 7.42-7.32 (m, 4H, H.sub.Ar), 6.87 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B), 6.64 (d, 1H, J.sub.2,NH=8.0 Hz, NH.sub.A), 5.90-5.80 (m, 1H, CH.sub.All), 5.29-5.23 (m, 1H, CH.sub.2All), 5.20-5.17 (m, 1H, CH.sub.2All), 4.93 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.88 (brs, 1H, H-1.sub.A), 4.83 (dd, 2H, J=12.4 Hz, J=7.6 Hz, CH.sub.2Nap), 4.72 (dd, 1H, J=3.6 Hz, J=10.2 Hz, H-3.sub.B), 4.67 (d, 1H, J=12.4 Hz, CH.sub.2Bn), 4.50-4.44 (m.sub.po, 2H, H-5.sub.A, CH.sub.2Bn), 4.39-4.31 (m, 2H, H-2.sub.A, CH.sub.2All), 4.08-4.03 (m, 1H, CH.sub.2All), 3.93-3.89 (m, 2H, H-3.sub.A, H-6a.sub.A), 3.82-3.75 (m, 3H, H-4.sub.B, H-5.sub.B, H-6b.sub.A), 3.53-3.44 (m, 2H, H-2.sub.B, H-4.sub.A), 1.40 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 162.3, 161.1 (CO.sub.NHTCA), 138.1, 134.4, 133.4, 133.1, 133.0 (C.sub.q,Ar), 133.4 (CH.sub.All), 128.5, 128.4, 128.3, 127.8, 127.7, 127.2, 126.3, 126.2, 125.7 (12C, C.sub.Ar), 118.0 (CH.sub.2All), 100.1 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 97.2 (C-1.sub.B, .sup.1J.sub.C,H=164 Hz), 92.3, 91.8 (2C, CCl.sub.3), 75.3 (C-3.sub.B), 72.6 (C-3.sub.A), 71.9 (CH.sub.2Nap), 70.5 (CH.sub.2Bn), 70.4 (C-4.sub.A), 70.2 (CH.sub.2All), 69.7 (C-5.sub.B), 69.0 (C-5.sub.A), 65.6 (C-4.sub.B), 62.2 (C-6.sub.A), 56.2 (C-2.sub.B), 51.4 (C-2.sub.A), 17.4 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.37H.sub.41Cl.sub.6N.sub.6O.sub.9, 925.1217; found 925.1223.

    [0597] Allyl (benzyl 3-O-benzyl-4-O-(2-naphthylmethyl)-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (6b). TEMPO (1.29 g, 8.26 mmol, 0.3 equiv.) was added, followed by BAIB (18.6 g, 57.8 mmol, 2.5 equiv.), to a biphasic solution of alcohol 5b (25.0 g, 3.7 mmol, 1.0 equiv.) in DCM/H.sub.2O (2:1, 690 mL). The biphasic mixture stirred vigorously for 6 h at rt, at which point a TLC analysis (Tol/EtOAc 1:4) revealed the absence of alcohol 5b (R.sub.f 0.65) and the presence of a polar product. NaHCO.sub.3 (9.7 g, 115 mmol, 4.2 equiv.) was added in 100 mL water. The DCM layer was separated, and the aq. phase was extracted with DCM (100 mL) twice. The combined organic phases were dried by passing through a phase separator filter and concentrated to dryness. The residue was dissolved in anhyd. DMF (250 mL). Benzyl bromide (4.25 mL, 35.8 mmol, 1.3 equiv.) and K.sub.2CO.sub.3 (4.95 g, 35.8 mmol, 1.3 equiv.) were added and the suspension was stirred at rt for 2 h. At completion, satd aq. NH.sub.4Cl was added and the aq. layer was washed with DCM (400 mL) thrice. The organic phases were combined, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (cHex/EtOAc 77:23.fwdarw.70:30) to give the desired benzyl ester 6b (23.0 g, 25.3 mmol, 82%) as a white-brown solid. Uronate 6b had R.sub.f 0.7 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.85-7.75 (m, 3H, HA), 7.69 (brs, 1H, HA), 7.51-7.47 (m, 2H, H.sub.Ar), 7.41-7.76 (m, 1H, H.sub.Ar), 6.77 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.64 (d, 1H, J.sub.2,NH=7.6 Hz, NH.sub.A), 5.90-5.80 (m, 1H, CH.sub.All), 5.29-5.17 (m, 5H, H-1.sub.A, CH.sub.2Bn-6, CH.sub.2All), 4.85 (d, 1H, J.sub.4,5=5.2 Hz, H-5.sub.A), 4.82 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.73 (brs, 2H, CH.sub.2Nap), 4.62 (dd, 1H, J.sub.3,4=4.0 Hz, J.sub.2,3=10.2 Hz, H-3.sub.B), 4.56 (d, 1H, J=12.0 Hz, CH.sub.2Bn), 4.48 (d, 1H, CH.sub.2Bn), 4.35-4.30 (m, 1H, CH.sub.2All), 4.23-4.18 (m, 1H, H-2.sub.A), 4.09-4.02 (m, 2H, H-4.sub.A, CH.sub.2All), 3.87-3.84 (m, 2H, H-4.sub.B, H-3.sub.A), 3.61-3.54 (m, 2H, H-2.sub.B, H-5.sub.B) 1.28 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.9 (C-6.sub.A), 162.0, 161.6 (CO.sub.NHTCA), 137.4, 135.0, 134.6, 133.1 (2C) (C.sub.q,Ar), 133.5 (CH.sub.All), 128.7, 128.6, 128.4, 128.3, 128.1, 127.9 (2C), 127.7, 127.0, 126.2, 126.1, 125.8 (12C, C.sub.Ar), 117.9 (CH.sub.2All), 98.9 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 97.7 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.3, 92.1 (2C, CCl.sub.3), 75.9 (C-3.sub.B), 73.4 (C-3.sub.A), 72.1 (C-4.sub.A), 72.0 (CH.sub.2Nap), 71.9 (CH.sub.2Bn), 71.6 (C-5.sub.A), 70.1 (CH.sub.2All), 69.4 (C-5.sub.B), 67.5 (CH.sub.2Bn-6), 65.2 (C-4.sub.B), 55.5 (C-2.sub.B), 53.1 (C-2.sub.A), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.44H.sub.47Cl.sub.6N.sub.6O.sub.10, 1029.1491; found 1029.1479.

    [0598] Allyl (benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (7)..sup.[1] Disaccharide 6b (11.0 g, 10.8 mmol, 1.0 equiv.) was dissolved in DCM (200 mL) and phosphate buffer pH 7 (20 mL) was added. The biphasic mixture was cooled to 0 C. and DDQ (7.4 g, 2.1 mmol, 3.0 equiv.) was added. The reaction was slowly allowed to reach rt and stirred for 6 h at this temperature. At completion, 10% aq. NaHCO.sub.3 (200 mL) was added and the biphasic mixture was diluted with DCM (200 mL). The DCM layer was separated, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (cHex/EtOAc 70:30.fwdarw.60:40) to give alcohol 7 (8.6 g, 8.50 mmol, 90%) as a white solid. Disaccharide 7 had R.sub.f 0.45 (Tol/EtOAc, 7:3). .sup.1H NMR (CDCl.sub.3) 7.42-7.25 (m, 10H, H.sub.Ar), 6.75 (d.sub.po, 1H, J.sub.2,NH=8.8 Hz, NH.sub.A), 6.73 (d.sub.po, 1H, J.sub.2,NH=7.6 Hz, NH.sub.B), 5.89-5.80 (m, 1H, CH.sub.All), 5.31-5.24 (m, 3H, CH.sub.2Bn-6, CH.sub.2All), 5.20-5.17 (m, 1H, CH.sub.2All), 4.98 (d, 1H, J.sub.1,2=3.2 Hz, H-1.sub.A), 4.83 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.72 (m, 3H, H-5.sub.A, CH.sub.2Bn), 4.64 (dd, 1H, J.sub.3,4=4.0 Hz, J.sub.2,3=10.6 Hz, H-3.sub.B), 4.35-4.27 (m, 1H, H-2.sub.A, CH.sub.2All), 4.15 (dt, 1H, J.sub.4,5=7.6 Hz, H-4.sub.A), 4.07-4.02 (m, 2H, CH.sub.2All), 3.88 (dd, 1H, J.sub.2,3=5.6 Hz, J.sub.3,4=3.6 Hz, H-3.sub.A), 3.79 (d, 1H, H-4.sub.B), 3.64 (q.sub.po, 1H, H-5.sub.B), 3.52-3.46 (m, 1H, H-2.sub.B), 2.78 (d, 1H, J.sub.4,OH=7.6 Hz, OH), 1.26 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.8 (C-6.sub.A), 162.1, 161.6 (CO.sub.NHTCA), 137.2, 135.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 128.7 (2C), 128.6, 128.4, 128.2, 125.2 (10C, C.sub.Ar), 118.0 (CH.sub.2All), 99.5 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 97.4 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.1, 91.9 (2C, CCl.sub.3), 76.3 (C-3.sub.B), 74.8 (C-3.sub.A), 72.4 (CH.sub.2Bn), 71.1 (C-5.sub.A), 70.1 (CH.sub.2All), 69.5 (C-5.sub.B), 67.5 (CH.sub.2Bn-6), 65.5 (C-4.sub.A), 65.5 (C-4.sub.B), 55.7 (C-2.sub.B), 51.4 (C-2.sub.A), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.33H.sub.39Cl.sub.6N.sub.6O.sub.10, 889.0853; found 889.0860.

    [0599] (Benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranose (9a). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (286 mg, 0.338 mmol, 0.03 equiv.) in anhyd. THF (20 mL) was degassed and stirred for 20 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly with Ar and poured into a solution of allyl glycoside 6b (11.4 g, 11.2 mmol, 1.0 equiv.) in anhyd. THF (200 mL). After stirring for 2 h at rt, a TCL follow up (Tol/EtOAc 4:1) revealed that the starting 6b (R.sub.f 0.6) had been converted to a closely migrating product (R.sub.f 0.65). NIS (3.0 g, 13.5 mmol, 1.2 equiv.) and H.sub.2O (20 mL) were added and after stirring for another 1 h at rt, 10% aq. Na.sub.2SO.sub.3 was added. The reaction mixture was concentrated and the aq. phase was extracted with DCM (200 mL) thrice. The combined organic layers were a washed with brine, dried over anhyd. Na.sub.2SO.sub.4, and concentrated under reduced pressure. The residue was purified by flash chromatography with cHex/EtOAc (70:30.fwdarw.60:40) to give the expected hemiacetal 9a (10.4 mg, 10.7 mmol, 95%) as a white solid. The / hemiacetal 9a had R.sub.f 0.3, 0.6 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.41H.sub.43Cl.sub.6N.sub.6O.sub.10, 989.1166; found 989.1177. The isomer had .sup.1H NMR (CDCl.sub.3) 7.87-7.76 (m, 3H, H.sub.Ar), 7.71 (brs, 1H, H.sub.Ar), 7.53-7.17 (m, 13H, H.sub.Ar), 7.06 (d, 1H, J.sub.2,NH=8.4 Hz, NH.sub.B), 6.86 (d, 1H, J.sub.2,NH=7.4 Hz, NH.sub.A), 5.40 (d, 1H, J.sub.1,2=6.4 Hz, H-1.sub.A), 5.28 (t, 1H, J.sub.1,2=3.5 Hz, H-1.sub.B), 5.19 (dd.sub.po, 2H, J=12.4 Hz, CH.sub.2Bn-6), 4.82 (d, 1H, J.sub.4,5=4.0 Hz, H-5.sub.A), 4.75 (brs, 2H, CH.sub.2Nap), 4.44-4.38 (m.sub.po, 3H, H-2.sub.B, CH.sub.2Bn), 4.26 (dd, 1H, J.sub.3,4=3.2 Hz, J.sub.2,3=10.6 Hz, H-3.sub.B), 4.22-4.09 (m.sub.po, 3H, H-2.sub.A, H-4.sub.A, H-5.sub.B), 3.95 (d, 1H, H-4.sub.B), 3.89 (dd, 1H, J.sub.2,3=2.8 Hz, J.sub.3,4=2.8 Hz, H-3.sub.A), 3.44 (brd, 1H, J.sub.4,OH=2.4 Hz, OH), 1.22 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.9 (C-6.sub.A), 162.0, 161.9 (CO.sub.NHTCA), 137.8, 137.0, 134.7, 134.5, 133.1 (2C) (C.sub.q,Ar), 129.0, 128.8 (2C), 128.7, 128.6, 128.5, 128.4 (2C), 128.3, 128.2, 128.1, 127.9, 127.7, 127.1, 126.2, 126.1, 126.0, 125.3 (10C, C.sub.Ar), 97.9 (C-1.sub.A, .sup.1J.sub.C,H=167 Hz), 92.4, 92.1 (2C, CCl.sub.3), 91.1 (C-1.sub.B, .sup.1J.sub.C,H=175 Hz), 75.2 (C-3.sub.B), 73.0 (C-3.sub.A), 72.4 (C-5.sub.A), 72.0 (C-4.sub.A), 72.0 (CH.sub.2Nap), 71.7 (CH.sub.2Bn), 67.6 (CH.sub.2Bn-6), 65.3 (C-5.sub.B), 65.2 (C-4.sub.B), 53.3 (C-2.sub.A), 51.0 (C-2.sub.B), 17.3 (C-6.sub.B).

    [0600] (Benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy-/-D-galactopyranosyl N-phenyltrifluoroacetimidate (9a) and 2-trichloromethyl [(benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-1,2,4,6-tetradeoxy--D-galactopyrano]-[2,1]-oxazoline (10a). PTFACl (721 L, 4.55 mmol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (1.25 g, 3.85 mmol, 1.1 equiv.) were added to a solution of hemiacetal 9a (3.4 g, 3.50 mmol, 1.0 equiv.) in acetone (70 mL). After stirring for 2 h at rt, the suspension was filtered over a pad of Celite and solids were washed with acetone (30 mL) twice. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography with cHex/EtOAc (90:1.fwdarw.80:20) to give a mix of the expected PTFA donor 9b and oxazoline 10b (3.6 g, 3.15 mmol, 91% calcd. wrt PTFA donor) as a white solid. Donor 9b had HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.49H.sub.47Cl.sub.6F.sub.3N.sub.7O.sub.10, 1160.1462; found 1160.1445. Oxazoline 10b had HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.41H.sub.38Cl.sub.6N.sub.5O.sub.9, 954.0795; found 954.0776.

    [0601] Oligomerization 2.sub.ANTCA, 2.sub.BNTCA (4.sub.A-Nap)

    ##STR00101##

    [0602] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (11b). PTFACl (1.52 mL, 9.63 mmol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (2.65 g, 8.15 mmol, 1.1 equiv.) were added to hemiacetal 9a (7.2 g, 7.41 mmol, 1.0 equiv.) in acetone (150 mL). After stirring for 2 h at rt, the reaction mixture was filtered through a pad of Celite and washed with DCM (50 mL) twice. The filtrate was concentrated under reduced pressure and dried under high vacuum to give the crude donors 9b/10b (8.4 g, 7.41 mmol, quant.), as a mix of PTFA donor 9b and oxazoline 10b. The crude material was used as such in the next step. Donors 9b/10b had R.sub.f 0.95 (Tol/EtOAc 4:1).

    [0603] A mix of the crude donors 9b/10b (8.4 g, 7.41 mmol, 1.06 equiv. theo.) and disaccharide acceptor 7 (6.0 g, 69.7 mmol, 1.0 equiv.) were co-evaporated with anhyd. toluene (50 mL) and then dried under vacuum. Freshly activated MS 4 (10 g) was added to the starting materials in anhyd. DCM (150 mL) and the suspension was stirred for 1 h under an Ar atmosphere at rt. After cooling to 15 C., TMSOTf (107 L, 593 mol, 0.08 equiv.) was added slowly and stirring went on for 45 min during which the bath temperature kept at 15 C. A TLC analysis (Tol/EtOAc 4:1) showed the absence of donors and the presence of a new spot in addition to a slight amount of less polar side products. At completion, Et.sub.3N (120 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (50 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 92:8.fwdarw.90:10) to give tetrasaccharide 11b as a white solid (9.1 g, 49.8 mmol, 71%). The coupling product 11b had R.sub.f 0.45 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.85-7.75 (m, 3H, H.sub.Ar), 7.69 (brs, 1H, H.sub.Ar), 7.43-7.20 (m, 26H, H.sub.Ar), 6.97 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B), 6.77 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.57 (d, 1H, J.sub.2,NH=7.6 Hz, NH.sub.A), 6.49 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.90-5.80 (m, 1H, CH.sub.All), 5.31-5.14 (m, 7H, H-1.sub.A, CH.sub.2Bn-6, CH.sub.2All), 5.05 (d, 1H, J.sub.1,2=5.2 Hz, H-1.sub.A), 4.85 (d.sub.po, 1H, J.sub.1,2=8.8 Hz, H-1.sub.B), 4.83 (d, 1H, J.sub.4,5=5.2 Hz, H-5.sub.A), 4.80 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.74-4.66 (m, 5H, H-3.sub.B, H-3.sub.B, H-5.sub.A, CH.sub.2Bn), 4.59 (d, 1H, J=12.0 Hz, CH.sub.2Bn), 4.48 (brd, 3H, CH.sub.2Bn, CH.sub.2Nap), 4.34-4.24 (m, 3H, H-2.sub.A, H-4.sub.A, CH.sub.2All), 4.08-4.01 (m, 2H, H-4.sub.A, CH.sub.2All), 3.93-3.88 (m, 3H, H-2.sub.A, H-3.sub.A, H-4.sub.B), 3.80-3.77 (m, 2H, H-3.sub.A, H-4.sub.B), 3.62-3.54 (m, 2H, H-2.sub.B, H-5.sub.B), 1.31 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B), 1.20 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.8, 168.2 (C-6.sub.A), 162.0, 161.9, 161.6, 161.5 (CO.sub.NHTCA), 137.4, 137.0, 135.1, 135.0, 133.4, 133.1 (2C) (C.sub.q,Ar), 133.5 (CH.sub.All), 129.1, 129.0, 128.7 (2C), 128.6, 128.5, 128.4, 128.3 (2C), 128.2, 128.0, 127.9, 127.7, 126.9, 126.2, 125.8, 125.2 (C.sub.Ar), 99.2 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 98.8 (C-1.sub.B, .sup.1J.sub.C,H=166 Hz), 98.1 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 97.7 (C-1.sub.B, .sup.1J.sub.C,H=166 Hz), 92.4, 92.3, 92.1 (4C, CCl.sub.3), 75.1 (C-3.sub.B), 74.7 (C-3.sub.B), 73.4 (C-3.sub.A), 73.0 (C-3.sub.A), 72.5 (CH.sub.2Nap), 72.0 (2C, C-4.sub.A), 71.9 (2C, CH.sub.2Bn), 71.5 (2C, C-5.sub.A), 70.0 (CH.sub.2All), 69.3 (C-5.sub.B), 68.8 (C-5.sub.B), 67.6 (CH.sub.2Bn-6), 67.4 (CH.sub.2Bn-6), 65.5 (C-4.sub.B), 65.1 (C-4.sub.B), 55.6 (C-2.sub.B), 55.5 (C-2.sub.B), 54.1 (C-2.sub.A), 52.7 (C-2.sub.A), 17.3 (C-6.sub.B). 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.74H.sub.76Cl.sub.12N.sub.11O.sub.19 1842.1576; found 1842.1593.

    [0604] Allyl (benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (15b). Tetrasaccharide 11b (4.0 g, 2.19 mmol, 1.0 equiv.) was dissolved in DCM (40 mL) and phosphate buffer pH 7 (4.0 mL) was added. The biphasic mixture was cooled to 0 C. and DDQ (996 mg, 4.38 mmol, 2.0 equiv.) was added. The reaction was stirred for 6 h, keeping the temperature between 0-10 C. At completion, 10% aq. NaHCO.sub.3 (100 mL) was added and the biphasic mixture was diluted with DCM (500 mL). The DCM layer was separated, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 75:25.fwdarw.70:30) to give alcohol 15b (3.2 g, 1.90 mmol, 86%) as a white solid. The tetrasaccharide acceptor 15b had R.sub.f 0.45 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.63H.sub.68Cl.sub.12N.sub.11O.sub.19 1702.0950; found 1702.0950.

    [0605] (Benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy-/-D-galactopyranose (12b). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (185 mg, 219 mol, 0.05 equiv.) in anhyd. THF (9.0 mL) was degassed and stirred for 30 min under an H.sub.2 atmosphere. The resulting deep yellow solution was degassed repeatedly with Ar and poured into a solution of allyl glycoside 11b (8.0 g, 4.38 mmol, 1.0 equiv.) in anhyd. THF (80 mL). After stirring for 2 h at rt, a TCL follow up (Tol/EtOAc 4:1, 2 runs) revealed that the starting 11b (R.sub.f 0.15) had been converted to a closely migrating product (R.sub.f 0.2). NIS (1.18 g, 5.26 mmol, 1.2 equiv.) and H.sub.2O (18 mL) were added and after stirring for another 1 h at rt, 10% aq. Na.sub.2SO.sub.3 was added. The reaction mixture was concentrated and the aq. phase was extracted with DCM (90 mL) thrice. The combined organic layers were washed with brine, dried over anhyd. Na.sub.2SO.sub.4, and concentrated under reduced pressure. The residue was purified by flash chromatography (cHex/EtOAc 80:20.fwdarw.75:25) to give the expected hemiacetal 12b (7.6 g, 7.51 mmol, 97%) as a white solid. The / hemiacetal 12b had R.sub.f 0.1, 0.15 (Tol/EtOAc 4:1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.71H.sub.72Cl.sub.12N.sub.11O.sub.19 1802.1263; found 1802.1242.

    [0606] (Benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy-/-D-galactopyranosyl (N-phenyl)trifluoroacetimidate (13b) and 2-trichloromethyl [(benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-1,2,4,6-tetradeoxy--D-galactopyrano]-[2,1]-oxazoline (14b). PTFACl (878 L, 5.53 mmol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (1.52 g, 4.68 mmol, 1.1 equiv.) were added to a solution of hemiacetal 12b (7.6 g, 4.26 mol, 1.0 equiv.) in acetone (85 mL). After stirring for 2 h at rt, the suspension was filtered over a pad of Celite and solids were washed with acetone (20 mL) twice. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography (cHex/EtOAc 85:15.fwdarw.80:20) to give the expected PTFA tetrasaccharide 13b in admixture with the corresponding oxazoline 14b (7.5 g, 7.51 mmol, 91%, calcd. wrt PTFA donor) as a white solid. Imidate 13b had HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.79H.sub.76Cl.sub.12N.sub.12O.sub.19 1973.1559; found 1973.1556. Oxazoline 14b had HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.71H.sub.70Cl.sub.12N.sub.11O.sub.18, 1784.1157; found 1784.1094.

    [0607] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-napthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (16b). [4+2]-glycosylation: A mix of donors 9b/10b (678 mg, 594 mol, 1.0 equiv.) and acceptor 15b (1.0 g, 594 mol, 1.0 equiv.) were co-evaporated with anhyd. toluene and then dried under vacuum for 1 h. Freshly activated 4 MS (1.0 g) was added to the starting materials in anhyd. DCE (15 mL) and the suspension was stirred for 45 min under an Ar atmosphere at rt. After cooling to 0 C., TMSOTf (8.6 L, 48 mol, 0.08 equiv.) was added slowly and stirring went on for 45 min during which the bath temperature was kept at 0 C. A TLC analysis (Tol/EtOAc 4:1) showed the absence of donors and the presence of a new spot. At completion, Et.sub.3N (10 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (20 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 92:8.fwdarw.86:14) to give hexasaccharide 16b as a white solid (1.15 g, 436 mol, 73%). The coupling product 16b had R.sub.f 0.2 (Tol/EtOAc 4:1) HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.104H.sub.105Cl.sub.18N.sub.16O.sub.28 2665.1580; found 2665.1580.

    [0608] Allyl (benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (17b). DDQ (155 mg, 683 mol, 3.0 equiv.) was added to hexasaccharide 16b (600 mg, 228 mol, 1.0 equiv.) in DCM (20 mL) and phosphate buffer pH 7 (2.0 mL). The biphasic mixture was cooled to 0 C. and stirred for 6 h while keeping the temperature between 0-10 C. A TLC analysis (Tol/EtOAc 3:1) showed the absence of the fully protected 16b (R.sub.f 0.6) and the presence of a more polar spot (R.sub.f 0.4). 10% Aq. NaHCO.sub.3 (10 mL) was added followed by DCM (20 mL). The DCM layer was separated, washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 80:20.fwdarw.70:30) to give alcohol 17b (490 mg, 196 mol, 86%) as a white solid. The hexasaccharide acceptor 17b had R.sub.f 0.2 (Tol/EtOAc 4:1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.93H.sub.97Cl.sub.18N.sub.16O.sub.28 2525.0942; found 2525.0949.

    [0609] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-napthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (18b). [4+4]-glycosylation: Donors 13b/14b (1.21 g, 621 mol, 1.0 equiv.) and acceptor 15b (1.04 g, 621 mol, 1.0 equiv.) were co-evaporated with anhyd. toluene and then dried under vacuum for 1 h. Freshly activated 4 MS (2.0 g) was added to the starting materials in anhyd. DCE (15 mL) and the suspension was stirred for 45 min under an Ar atmosphere at rt. After cooling to 0 C., TMSOTf (10 L, 565 mol, 0.09 equiv.) was added slowly and stirring went on for 45 min during which the bath temperature was kept at 0 C. A TLC analysis (Tol/EtOAc 4:1) showed the absence of donors 13b/14b and the presence of a new spot. At completion, Et.sub.3N (15 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (20 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 88:12.fwdarw.84:16) to give octasaccharide 18b as a white solid (1.5 g, 434 mol, 70%). The coupling product 18b had R.sub.f 0.55 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.134H.sub.138Cl.sub.24N.sub.22O.sub.37 1749.0997; found 1749.0984.

    [0610] Allyl (benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (19b). DDQ (223 mg, 983 mol, 3.0 equiv.) was added to octasaccharide 18b (600 mg, 328 mol, 1.0 equiv.) in DCM (30 mL) and phosphate buffer pH 7 (3.0 mL). The biphasic mixture was cooled to 0 C. and stirred for 6 h while keeping the temperature between 0-10 C. A TLC analysis (Tol/EtOAc 7:3) showed the absence of the fully protected 18b (R.sub.f 0.6) and the presence of a more polar spot. 10% Aq. NaHCO.sub.3 (30 mL) was added followed by DCM (30 mL). The DCM layer was separated, washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 80:20.fwdarw.70:30) to give alcohol 19b (990 mg, 299 mol, 91%) as a white solid. The octasaccharide acceptor 19b had R.sub.f 0.35 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.123H.sub.129Cl.sub.24N.sub.22O.sub.37 1679.0681; found 1679.0663.

    [0611] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-napthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (20b). [4+6]-glycosylation: Tetrasaccharide donors 13b/14b (235 mg, 120 mol, 1.0 equiv.) and acceptor 17b (300 mg, 120 mol, 1.0 equiv.) were co-evaporated with anhyd. toluene and then dried under vacuum. Freshly activated 4 MS (500 mg) was added to the starting materials in anhyd. DCE (10 mL) and the suspension was stirred for 45 min under an Ar atmosphere at rt. After cooling to 0 C., TMSOTf (2 L, 11 mol, 0.09 equiv.) was added slowly and stirring went on for 45 min during which the bath temperature kept at 0 C. A TLC analysis (Tol/EtOAc 4:1) showed the absence of donor 13b/14b and the presence of a new spot. At completion, Et.sub.3N (2 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (20 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 84:16.fwdarw.82:18) to give decasaccharide 20b as a white solid (330 mg, 77.4 mol, 64%). The coupling product 20b had R.sub.f 0.5 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.164H.sub.175Cl.sub.30N.sub.27O.sub.46 2157.6027; found 2157.6028.

    [0612] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-napthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.-3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (21b). [4+8]-glycosylation: The tetrasaccharide donors 13b/14b (490 mg, 251 mol, 1.0 equiv.) and the octasaccharide acceptor 17b (830 mg, 251 mol, 1.0 equiv.) were co-evaporated with anhyd. toluene and then dried under vacuum. Freshly activated 4 MS (1.0 g) was added to the starting materials in anhyd. DCE (15 mL) and the suspension was stirred for 1 h under an Ar atmosphere at rt. After cooling to 0 C., TMSOTf (4.1 L, 23 mol, 0.09 equiv.) was added slowly and stirring went on for 45 min during which the bath temperature kept at 0 C. A TLC analysis (Tol/EtOAc 7:3) showed the absence of donors 13b/14b and the presence of a new spot. At completion, Et.sub.3N (5 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (15 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 82:18.fwdarw.78:22) to give dodecasaccharide 21b as a white solid (900 mg, 177 mol, 70%). The coupling product 21b had R.sub.f 0.45 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.194H.sub.196Cl.sub.36N.sub.32O.sub.55 2566.1056; found 2566.1059.

    [0613] Full Deprotection from the Fully Protected Precursors Featuring a 4.sub.A-Nap

    ##STR00102##

    [0614] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (1). Disaccharide 6b (140 mg, 182 mol) was subjected to deprotection (protocol 1). The known propyl glycoside 1 was obtained as a white lyophilized powder (66 mg, 130 mol, 78%). The target propyl glycoside 1 had RP-HPLC (215 nm): R.sub.t=12.3 min (conditions A), R.sub.t=13.9 min (conditions B), R.sub.t=12.2 min (conditions C). Analytical data were as above.

    [0615] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (2). Tetrasaccharide 11b (110 mg, 65 mol) was subjected to hydrogenation (protocol 1). The desired propyl glycoside was obtained as a white lyophilized powder (28 mg, 32 mol, 49%). Tetrasaccharide 2 had RP-HPLC (215 nm): R.sub.t=12.3 min (conditions A), R.sub.t=13.6 min (conditions B). Analytical data were as above.

    [0616] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (3). The fully protected hexasaccharide 16b (100 mg, 38 mol) was subjected to hydrogenation (protocol 1). The desired propyl glycoside 3 was obtained as a white lyophilized solid (19 mg, 15 mol, 39%). The free hexasaccharide 3 had RP-HPLC (215 nm): R.sub.t=11.3 min (conditions A), R.sub.t=13.4 min (conditions B). Analytical data were as above.

    [0617] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (4). The fully protected octasaccharide 18b (20 mg, 6 mol) was subjected to deprotection (protocol 2). A solution of the starting material in iPrOH/MeTHF/H.sub.2O (10:1:1) was passed through a 20% Pd(OH).sub.2C cartridge at a flow of 0.8 mL/min in the full H.sub.2 mode. After the first run was over, 0.12 mM aq. NaHCO.sub.3 (50 L, 6 mol, 1 equiv.) was added. And the same was repeated over the next five runs to reach a total of 5 equiv. of NaHCO.sub.3. The product was obtained as a white solid (4.9 mg, 2.9 mol, 50%). The free octasaccharide 4 had RP-HPLC (215 nm): R.sub.t=13.3 min (conditions A).

    [0618] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (22b). The fully protected decasaccharide 20b (30 mg, 7.0 mol) was subject to hydrogenation (protocol 2). The product was obtained as a white lyophilized foam (4.9 mg, 2.4 mol, 33%). The free decasaccharide 22b had RP-HPLC (215 nm): R.sub.t=13.5 min (conditions A). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.83H.sub.135N.sub.15O.sub.46 1038.9337; found 1038.9330.

    [0619] Propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (23b). The fully protected dodecasaccharide 21b (50 mg, 10.7 mol) was subjected to one step hydrogenation-mediated final deprotection (protocol 2). The free propyl glycoside was isolated as a white lyophilized material (1.5 mg, 0.61 mol, 6%). The free dodecasaccharide 23b had RP-HPLC (215 nm): R.sub.t=14.1 min (conditions A). HRMS (ESI.sup.+): m/z [M+3H].sup.3+ calcd for C.sub.99H.sub.161N.sub.18O.sub.55 827.6790; found 827.6793.

    Example 7: Strategy 2.SUB.A.-NTCA,2.SUB.B.-NTCA, Featuring a 4.SUB.A.-Me Endchain Disaccharide

    [0620] The Chain Terminator 4.sub.A-Me AB Disaccharide Building Block

    ##STR00103## ##STR00104##

    [0621] Allyl 2-azido-3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-methyl--L-altropyranoside (31b). CSA (1.3 g, 5.90 mmol, 0.5 equiv.) was added to the benzylidene acetal 12 (5.0 g, 11.8 mmol, 1.0 equiv.) in MeOH/DCM (4:1, 50 mL). After stirring at rt for 2 h, a TLC follow up (Tol/EtOAc 4:1) indicated reaction completion as shown by the absence of the starting material (R.sub.f 0.65) and the presence of a polar spot. 10% Aq. NaHCO.sub.3 (100 mL) was added followed by EtOAc (100 mL). The organic phase was separated and washed with brine (100 mL). The organic phase was dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude product was dried under high vacuum. tert-Butyldiphenylchlorosilane (3.37 mL, 12.9 mmol, 1.1 equiv.) and imidazole (1.6 g, 23.6 mmol, 2.0 equiv.) were added to the crude diol intermediate in anhyd. DMF (60 mL) at 0 C. The reaction mixture was allowed to reach rt slowly and stirred overnight at this temperature. Methanol (5.0 mL) was added and after 30 min, volatiles were evaporated under reduced pressure. The residue was dissolved in EtOAc and the organic layer was washed with 90% aq. brine, separated, dried over Na.sub.2SO.sub.4, and concentrated to provide the crude 6-O-silylated intermediate. Mel (1.47 g, 23.6 mmol, 2.0 equiv.) was added to the crude intermediate in DMF (230 mL). The solution was cooled to 0 C. and NaH (60% in mineral oil, 567 mg, 23.6 mmol, 2.0 equiv.) was added portionwise. After stirring vigorously for 2 h while the bath temperature slowly reached rt, a TLC follow up indicated reaction completion. The reaction mixture was diluted with DCM (1 L) and 5% aq. NH.sub.4Cl (100 mL) was added. The organic layer was washed with water and brine, dried over Na.sub.2SO.sub.4 and concentrated. The crude product was purified by flash chromatography (cHex/EtOAc 92:8.fwdarw.88:12) to give the fully protected 31b (6.9 g, 11.7 mmol, 99%) as a light yellow oil. Allyl glycoside 31b had R.sub.f 0.65 (Tol/EtOAc 10:1). .sup.1H NMR (CDCl.sub.3) 7.71-7.68 (m, 4H, H.sub.Ar), 7.47-7.31 (m, 11H, H.sub.Ar), 5.95-5.90 (m, 1H, CH.sub.All), 5.34-5.29 (m, 1H, CH.sub.2All), 5.21-5.18 (m, 1H, CH.sub.2All), 4.76-4.67 (m, 3H, H-1, CH.sub.2Bn), 4.30-4.25 (m, 1H, CH.sub.2All), 4.12 (q, 1H, J.sub.5,6b=4.8 Hz, H-5), 4.07-4.02 (m, 1H, CH.sub.2All), 3.90 (dd, 1H, J.sub.1,2=4.8 Hz, J.sub.2,3=8.0 Hz, H-2), 3.81 (brd, 2H, J.sub.5,6=4.4 Hz, H-6a, H-6b), 3.77 (dd, 1H, J.sub.3,4=3.6 Hz, H-3), 3.66 (dd, 1H, J.sub.4,5=5.2 Hz, H-4), 3.39 (s, 3H, OCH.sub.3), 1.09 (s, 9H, CH.sub.3,TBDPS). .sup.13C NMR (CDCl.sub.3) 137.8, 133.2, 133.0 (C.sub.q,Ar), 133.8 (CH.sub.All), 135.7, 135.5, 129.8, 128.4, 127.8, 127.7 (2C), 127.5 (C.sub.Ar), 117.3 (CH.sub.2All), 98.7 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 75.8 (C-3), 74.9 (C-4), 72.5 (C-5), 72.3 (CH.sub.2Bn), 68.7 (CH.sub.2All), 63.9 (C-6), 61.7 (C-2), 58.2 (OCH.sub.3), 26.8 (CH.sub.3,TBDPS), 19.2 (C.sub.TBDPS). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.33H.sub.45N.sub.4O.sub.5Si, 605.3154; found 605.3153.

    [0622] Allyl 3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranoside (32b). Zn dust (7.6 g, 117 mmol, 10.0 equiv.) and AcOH (6.7 mL, 117 mmol, 10.0 equiv.) were added to azide 31b (6.9 g, 11.7 mmol, 1.0 equiv.) in anhyd. THF (120 mL). After stirring for 1 h at rt, a TLC analysis (Tol/EtOAc 10:1) showed the absence of azide 31b (R.sub.f 0.8) and the presence of a more polar spot. The suspension was filtered over a pad of Celite and washed with THF. The filtrate was concentrated and then diluted with DCM. The DCM layer was washed with satd aq. NaHCO.sub.3, water, and brine, dried over Na.sub.2SO.sub.4, concentrated under reduced pressure, and dried under high vacuum. Trichloroacetyl chloride (1.97 mL, 17.6 mmol, 1.5 equiv.) and triethylamine (3.25 mL, 23.4 mmol, 2.0 equiv.) were added to a solution of the crude amine in DCM (120 mL). After stirring at rt for 1 h, a TLC follow up (Tol/EtOAc 10:1) indicated reaction completion. Methanol (2 mL) was added and the reaction mixture was stirred for another 10 min. Volatiles were evaporated and dried under high vacuum. The crude was taken in DCM (300 mL) and the DCM layer was washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (cHex/EtOAc 94:6.fwdarw.90:10) to give the fully protected 32b (6.95 g, 118 mmol, 83%) as a yellowish dense oil. Allyl glycoside 32b had R.sub.f 0.7 (Tol/EtOAc 10:1). .sup.1H NMR (CDCl.sub.3) 7.75-7.71 (m, 4H, H.sub.Ar), 7.47-7.29 (m, 11H, H.sub.Ar), 6.84 (d, 1H, J.sub.2,NH=8.4 Hz, NH), 5.95-5.86 (m, 1H, CH.sub.All), 5.34-5.29 (m, 1H, CH.sub.2All), 5.20-5.17 (m, 1H, CH.sub.2All), 4.88 (d.sub.po, 1H, J=12.0 Hz, CH.sub.2Bn), 4.87 (d, 1H, J.sub.1,2=4.8 Hz, H-1), 4.70 (d, 1H, CH.sub.2Bn), 4.48 (ddd, 1H, J.sub.2,3=3.6 Hz, H-2), 4.27 (m, 2H, H-5, CH.sub.2All), 4.06-3.98 (m.sub.po, 2H, H-6a, CH.sub.2All), 3.90 (dd, 1H, J.sub.5,6b=2.4 Hz, J.sub.6a,6b=11.2 Hz, H-6b), 3.67 (dd, 1H, J.sub.4,5=9.2 Hz, H-4), 3.27 (s, 3H, OCH.sub.3), 1.09 (s, 9H, CH.sub.3,TBDPS). .sup.13C NMR (CDCl.sub.3) 161.2 (CO.sub.NHTCA), 137.9, 133.5, 133.3 (C.sub.q,Ar), 133.5 (CH.sub.All), 135.7, 133.5, 129.6, 128.2, 128.1, 127.6 (15C, C.sub.Ar), 117.3 (CH.sub.2All), 97.3 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 73.0 (C-4), 71.5 (CH.sub.2Bn), 71.2 (C-3), 68.5 (C-5), 68.3 (CH.sub.2All), 62.8 (C-6), 56.4 (OCH.sub.3), 50.8 (C-2), 26.8 (CH.sub.3,TBDPS), 19.4 (C.sub.TBDPS). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.35H.sub.46Cl.sub.3N.sub.2O.sub.6Si, 723.2185; found 723.2184.

    [0623] 3-O-Benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranose (33b). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (248 mg, 294 mol, 0.03 equiv.) was dissolved in anhyd. THF (10 mL) and stirred for 30 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed several times and poured into a solution of allyl glycoside 32b (6.9 g, 9.78 mmol, 1.0 equiv.) in anhyd. THF (90 mL) under Ar. After stirring for 2 h at rt, a solution of NaHCO.sub.3 (2.4 g, 29.3 mmol, 3.0 equiv) in H.sub.2O (10 mL) was added, followed by a solution of iodine (4.96 g, 19.5 mmol, 2.0 equiv) in THF (10 mL). After stirring for 1 h at rt, the reaction was quenched with 10% aq. Na.sub.2SO.sub.3. The reaction mixture was concentrated and the aq. phase was extracted with DCM (3100 mL). The organic phases were pooled, washed with brine, dried over anhyd. Na.sub.2SO.sub.4 and concentrated. Purification by flash chromatography (cHex/EtOAc 90:10.fwdarw.88:12) gave hemiacetal 33b (/-4:1, 5.8 g, 8.71 mmol, 89%) as a white foam. The /-anomer 33b had R.sub.f 0.55, 0.5 (Tol/EtOAc 4:1); .sup.1H NMR (CDCl.sub.3) 7.75-7.69 (m, 5.2H, H.sub.Ar), 7.47-7.26 (m, 14.4H, H.sub.Ar), 7.21-7.18 (m, 0.56H, H.sub.Ar), 7.09 (d, 1H, J.sub.2,NH=6.4 Hz, NH.sub.), 6.84 (d, 0.24H, J.sub.2,NH=7.6 Hz, NH.sub.), 5.61 (d, 0.24H, J.sub.1,OH=11.2 Hz, OH.sub.), 5.37 (dd, 1H, H-1.sub.), 5.10 (dd.sub.po, 0.24H, H1.sub.), 4.90-4.75 (m, 2.54H, CH.sub.2Bn,, CH.sub.2Bn,), 4.39-4.36 (m, 0.51H, H-2.sub., H-5), 4.31-4.25 (m, 2H, H-2.sub., H-5.sub.), 4.13-4.07 (m, 0.54H, H-6.sub., H-4), 3.98-3.88 (m, 3.3H, H-4.sub., H-6ab.sub.,, H-6b.sub.), 3.80 (dd, 0.28H, J.sub.2,3=2.4 Hz, J.sub.3,4=9.6 Hz, H-3), 3.65 (dd, 1H, J.sub.2,3=3.2 Hz, J.sub.3,4=9.2 Hz, H-3.sub.), 3.32 (s, 0.76H, OCH.sub.3,), 3.29 (s, 3H, OCH.sub.3,), 3.06 (d, 1H, J.sub.1,OH=4.0 Hz, OH.sub.), 1.09, 1.08 (2s, 11.6H, CH.sub.3,TBDPS). .sup.13C NMR (CDCl.sub.3) 162.2, 161.4 (CO.sub.NHTCA), 137.8, 136.5, 135.6, 135.5 (2C), 133.6, 133.3, 133.2, 133.1 (C.sub.q,Ar), 135.7, 135.6, 135.5 (2C), 129.7 (2C), 129.0, 128.7, 128.5, 128.4, 128.3, 128.0, 127.9, 127.7, 127.6, 125.3 (C.sub.Ar), 93.4 (C-1.sub., .sup.1J.sub.C,H=174 Hz), 91.3 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 74.3 (CH.sub.2Bn,), 74.1 (C-4.sub.), 73.5 (C-3.sub.), 73.0 (CH.sub.2Bn,), 72.8 (C-3.sub.), 72.7 (C-5.sub.), 71.3 (C-5.sub.), 67.6 (C-4.sub.), 62.9 (C-6.sub.), 62.9 (C-6.sub.), 57.1 (OCH.sub.3), 53.1 (C-2.sub.), 51.3 (C-2.sub.), 26.9, 26.8 (CH.sub.3, TBDPS), 19.4 (C.sub.TBDPS). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.32H.sub.42Cl.sub.3N.sub.2O.sub.6Si, 683.1872; found 683.1876.

    [0624] Allyl 3-O-benzyl-6-O-tert-butyldiphenylsilyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyl-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (35b). Hemiacetal 33b (5.8 g, 8.72 mmol, 1.0 equiv.) was dissolved in acetone (80 mL). PTFACl (1.79 mL, 11.3 mmol, 1.3 equiv.) was added followed by Cs.sub.2CO.sub.3 (3.4 mg, 10.4 mmol, 1.2 equiv). After stirring for 2 h at rt, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The PTFA donor 34b had R.sub.f 0.8 (Tol/EtOAc 4:1).

    [0625] The crude PTFA donor 34b (7.29 g, 8.72 mmol, 1.0 equiv. theo.) and acceptor 8 (3.2 g, 8.72 mmol, 1.0 equiv) were co-evaporated with anhyd. toluene (40 mL) and dried under vacuum. The dried mass was dissolved in anhyd. DCE (120 mL), freshly activated 4 MS (5.0 g) was added and the suspension was stirred for 30 min at rt under an Ar atmosphere. The reaction mixture was cooled to 15 C. and TMSOTf (79 L, 436 mol, 0.05 equiv.) was added. After 40 min at 15 C., a TLC analysis (Tol/EtOAc 4:1) showed the absence of donor and the presence of a new spot. The mixture was quenched with Et.sub.3N, filtered and concentrated. Flash chromatography (cHex/EtOAc 90:10.fwdarw.88:12) yielded the coupling product 35b (8.2 g, 8.04 mmol, 92%) as a white foam. The desired disaccharide 35b had R.sub.f 0.5 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.73-7.70 (m, 4H, H.sub.Ar), 7.52-7.28 (m, 11H, H.sub.Ar), 6.73 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.67 (d, 1H, J.sub.2,NH=8.0 Hz, NH.sub.A), 5.90-5.81 (m, 1H, CH.sub.All), 5.29-5.24 (m, 1H, CH.sub.2All), 5.21-5.17 (m, 1H, CH.sub.2All), 4.92 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.85 (d.sub.po, 1H, J.sub.1,2=4.4 Hz, H-1.sub.A), 4.84 (d.sub.po, 1H, J=12.4 Hz, CH.sub.2Bn), 4.77 (d, 1H, CH.sub.2Bn), 4.59 (dd, 1H J.sub.2,3=10.8 Hz, J.sub.3,4=4.0 Hz, H-3.sub.B), 4.41-4.30 (m, 3H, H-2.sub.A, H-5.sub.A, CH.sub.2All), 4.08-4.01 (m, 3H, H-4.sub.A, H-6a.sub.A, CH.sub.2All), 3.95 (dd, J.sub.5,6b=2.0 Hz, J.sub.6a,6b=11.2 Hz, H-6b.sub.A), 3.63 (d.sub.po, 1H, J.sub.4,5=3.6 Hz, H-4.sub.B), 3.60 (dq.sub.po, 1H, J.sub.4,5=1.0 Hz, H-5.sub.B), 3.53 (dd, 1H, J.sub.2,3=9.2 Hz, J.sub.3,4=3.6 Hz, H-3.sub.A), 3.43-3.36 (m, 1H, H-2.sub.B), 3.29 (s, 3H, OCH.sub.3), 1.23 (d, 3H, 6.4 Hz, H-6.sub.B), 1.09 (s, 9H, CH.sub.3,TBDPS). .sup.13C NMR (CDCl.sub.3) 162.2, 161.1 (CO.sub.NHTCA), 138.4, 133.4, 133.3 (C.sub.q,Ar), 133.5 (CH.sub.All), 135.7, 135.5, 129.8, 128.8, 128.2, 128.7 (C.sub.Ar), 117.8 (CH.sub.2All), 100.6 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 97.3 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 92.1, 92.0 (2C, CCl.sub.3), 76.2 (C-3.sub.B), 73.3 (C-3.sub.A), 72.3 (CH.sub.2Bn), 72.0 (C-4.sub.A), 70.1 (CH.sub.2All), 69.7 (C-5.sub.B), 69.4 (C-5.sub.A), 65.6 (C-4.sub.B), 63.2 (C-6.sub.A), 56.7 (OCH.sub.3), 56.0 (C-2.sub.B), 51.4 (C-2.sub.A), 26.9 (CH.sub.3,TBDPS), 19.4 (C.sub.TBDPS), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.43H.sub.55Cl.sub.6N.sub.6O.sub.9Si, 1037.1925; found 1037.1914.

    [0626] Allyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyl-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (36b). TBAF.Math.3H.sub.2O (2.91 g, 9.25 mmol, 1.15 equiv.) was added to disaccharide 35b (8.2 g, 8.0 mmol, 1.0 equiv.) in THF (80 mL) and the reaction mixture was stirred at rt for 2 h. A TLC analysis (EtOAc) showed the consumption of the protected disaccharide 35b (R.sub.f 1.0) and the presence of a polar spot. Acetic acid (0.53 mL, 9.25 mmol, 1.15 equiv.) was added slowly and after stirring for 10 min, volatiles were evaporated. The residue was purified by flash chromatography (cHex/EtOAc 30:70.fwdarw.0:100) to give alcohol 36b (5.8 g, 7.42 mmol, 92%) as a white solid. Disaccharide 36b had R.sub.f 0.35 (EtOAc). .sup.1H NMR (DMSO-d.sub.6) 8.92 (d, 1H, J.sub.2,NH=8.4 Hz, NH.sub.A), 8.82 (d, 1H, J.sub.2,NH=9.2 Hz, NH.sub.B), 7.37-7.24 (m, 5H, H.sub.Ar), 5.85-5.75 (m, 1H, CH.sub.All), 5.25-5.19 (m, 1H, CH.sub.2All), 5.11-5.08 (m, 1H, CH.sub.2All), 5.00 (d, 1H, J.sub.1,2=4.0 Hz, H-1.sub.A), 4.82 (t, 1H, J.sub.6,OH=5.6 Hz, OH), 4.61 (d, 1H, J=11.6 Hz, CH.sub.2Bn), 4.55 (d, 1H, CH.sub.2Bn), 4.48 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.22-4.14 (m, 3H, H-3.sub.B, H-2.sub.A, CH.sub.2All), 4.08-4.02 (m, 2H, H-4.sub.B, H-5.sub.A), 3.98-3.93 (m, 1H, CH.sub.2All), 3.86-3.79 (m, 2H, H-4.sub.A, H-2.sub.B), 3.69-3.60 (m.sub.po, 2H, H-5.sub.B, H-6a.sub.A), 3.58-3.54 (m.sub.po, 2H, H-3.sub.A, H-6b.sub.A), 3.24 (s, 3H, OCH.sub.3), 1.23 (d, 3H, 6.4 Hz, H-6.sub.B). .sup.13C NMR (DMSO-d.sub.6) 162.0, 161.8 (CO.sub.NHTCA), 138.8 (C.sub.q,Ar), 134.8 (CH.sub.All), 128.4, 128.0, 127.7 (C.sub.Ar), 116.9 (CH.sub.2All), 100.8 (C-1.sub.A, .sup.1J.sub.C,H=172 Hz), 100.4 (C-1.sub.B, .sup.1J.sub.C,H=161 Hz), 93.6, 93.2 (2C, CCl.sub.3), 76.3 (C-3.sub.B), 74.5 (C-3.sub.A), 74.0 (C-4.sub.A), 72.7 (C-5.sub.A), 71.4 (CH.sub.2Bn), 69.5 (CH.sub.2All), 69.3 (C-5.sub.B), 65.4 (C-4.sub.B), 61.7 (C-6.sub.A), 57.6 (OCH.sub.3), 53.4 (C-2.sub.B), 52.7 (C-2.sub.A), 17.7 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.27H.sub.37Cl.sub.6N.sub.6O.sub.9, 799.0748; found 799.0748.

    [0627] Allyl (benzyl 3-O-benzyl-4-O-methyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (37b). TEMPO (234 mg, 1.49 mmol, 0.2 equiv) and BAIB (6.0 g, 18.7 mmol, 2.5 equiv) were added to a solution disaccharide 36b (5.85 g, 7.49 mmol, 1.0 equiv.) in biphasic DCM/H.sub.2O (1:10, 150 mL). After stirring for 2 h at rt, the reaction was quenched with 10% aq. Na.sub.2SO.sub.3 and the biphasic mixture was diluted with DCM (150 mL). The aq. phase was separated and extracted twice with DCM (150 mL). The combined organic phases were washed with brine, dried by passing through a phase separator filter and concentrated under reduced pressure. To a solution of the crude material in anhyd. DMF (40 mL) were added benzyl bromide (2.66 mL, 22.4 mmol, 3.0 equiv) and K.sub.2CO.sub.3 (1.3 g, 9.73 mmol, 1.3 equiv.). After stirring for 1 h at rt, the reaction mixture was diluted with 10% aq. NH.sub.4Cl (100 mL) and extracted with DCM (200 mL) twice. The organic phases were pooled, washed with brine, dried over anhyd Na.sub.2SO.sub.4, filtered and concentrated in vacuo. Flash chromatography using Tol/EtOAc (4:1) gave the benzyl ester 37b (5.8 g, 6.55 mmol, 87%) as a white solid. Uronate 37b had R.sub.f 0.6 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.44-7.28 (m, 10H, H.sub.Ar), 6.78 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.73 (d, 1H, J.sub.2,NH=7.6 Hz, NH.sub.A), 5.88-5.80 (m, 1H, CH.sub.All), 5.30-5.23 (m.sub.po, 3H, CH.sub.2Bn-6, CH.sub.2All), 5.22 (d.sub.po, 1H, J.sub.1,2=6.0 Hz, H-1.sub.A), 5.19-5.16 (m, 1H, CH.sub.All), 4.82 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.78 (d, 1H, J.sub.4,5=5.2 Hz, H-5.sub.A), 4.65-4.53 (m, 3H, H-3.sub.B, CH.sub.2Bn), 4.34-4.29 (m, 1H, CH.sub.2All), 4.14-4.08 (m, 1H, H-2.sub.A), 4.06-4.01 (m, 1H, CH.sub.2All), 3.97 (dd, 1H, J.sub.2,3=8.4 Hz, J.sub.3,4=2.8 Hz, H-3.sub.A), 3.85-3.83 (m, 2H, H-4.sub.B, H-4.sub.A), 3.60-3.52 (m, 2H, H-2.sub.B, H-5.sub.B), 3.38 (s, 3H, OCH.sub.3), 1.27 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.9 (CO.sub.CO2Bn), 161.9, 161.7 (CO.sub.NHTCA), 137.4, 135.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 128.7 (3C), 128.4, 128.1, 128.0 (C.sub.Ar), 117.9 (CH.sub.2All), 98.7 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 97.7 (C-1.sub.B, .sup.1J.sub.C,H=164 Hz), 92.3, 92.2 (2C, CCl.sub.3), 75.8 (C-3.sub.B), 75.1 (C-4.sub.A), 73.2 (C-3.sub.A), 72.1 (CH.sub.2Bn), 71.4 (C-5.sub.A), 70.1 (CH.sub.2All), 69.3 (C-5.sub.B), 67.5 (CH.sub.2Bn-6), 65.2 (C-4.sub.B), 58.2 (OCH.sub.3), 55.5 (C-2.sub.B), 53.3 (C-2.sub.A), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.34H.sub.41Cl.sub.6N.sub.6O.sub.10, 903.1010; found 903.1014.

    [0628] (Benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranose (38b). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (149 mg, 0.176 mmol, 0.03 equiv) was dissolved in anhyd THF (10 mL) and stirred for 30 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed several times and poured into a solution of the allyl glycoside 31b (5.2 mg, 5.87 mmol, 1.0 equiv.) in anhyd THF (80 mL). After stirring for 2 h at rt, a solution of iodine (2.9 g, 11.7 mmol, 2.0 equiv) and NaHCO.sub.3 (1.48 g, 17.6 mmol, 3.0 equiv.) were added. After stirring for 2 h at rt, the reaction was quenched with 10% aq sodium sulphite. The reaction mixture was concentrated and the aq phase was extracted with DCM (3150 mL). The organic phases were pooled, washed with brine, dried over anhyd. Na.sub.2SO.sub.4 and concentrated. Purification by flash chromatography (Tol/EtOAc 75:25.fwdarw.70:30) yielded hemiacetal 38b (/ 3.5:1, 4.6 g, 5.44 mmol, 92%) as a white foam. The /-anomer had R.sub.f 0.2, 0.5 (Tol/EtOAc 7:3). .sup.1H NMR (CDCl.sub.3) 7.45-7.18 (m, 15H, HA, NH.sub.B-), 7.04 (d, 1H, J.sub.2,NH=8.4 Hz, NH.sub.B-), 6.87 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.A-), 6.83 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.A-), 5.43 (d, 1H, J.sub.1,2=6.8 Hz, H-1.sub.A-), 5.39 (d, 0.3H, J.sub.1,2=6.8 Hz, H-1.sub.A-), 5.29 (t.sub.po, 1.1H, J.sub.1,2=3.6 Hz, H-1.sub.B-), 5.6-5.21 (m.sub.po, 2.6H, CH.sub.2Bn-6), 4.81 (d, 0.3H, J.sub.4,5=3.6 Hz, H-5.sub.A-), 4.79 (d, 1H, J.sub.4,5=3.2 Hz, H-5.sub.A-), 4.67 (t, 0.3H, J.sub.1,2=8.8 Hz, H-1.sub.B), 4.61-4.37 (m, 2.7H, CH.sub.2Bn), 4.43-4.37 (m, 1H, H-2.sub.B-), 4.26 (dd, 1H, J.sub.2,3=8.4 Hz, J.sub.3,4=3.2 Hz, H-3.sub.B-), 4.17 (dd, 1H, J.sub.2,3=8.4 Hz, J.sub.3,4=3.2 Hz, H-3.sub.B-), 4.13-4.03 (m, 2.7H, H-5.sub.B-, H-2.sub.A-, H-2.sub.A-), 3.96-3.86 (m, 4H, H-4.sub.B-, H-4.sub.A-, H-3.sub.A-, H-2.sub.A-, H-3.sub.A-, H-4.sub.B-), 3.82 (dd, 0.3H, J.sub.3,4=2.8 Hz, J.sub.4,5=9.6 Hz, H-4.sub.A-), 3.62-3.59 (m, 0.3H, H-5.sub.B-), 3.54 (s, 1H, OCH.sub.3-), 3.41 (s, 1H, OCH.sub.3-), 3.07 (d, 1H, J.sub.1,OH=3.6 Hz, OH), 1.32 (d, 1H, J.sub.5,6=6.4 Hz, H-6.sub.B-) 1.27 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B-). .sup.13C NMR (CDCl.sub.3) 168.9, 168.7 (CO.sub.CO2Bn), 162.2, 162.0, 161.8 (CO.sub.NHTCA), 137.0, 136.8, 134.8 (C.sub.Ar), 129.0, 128.9, 128.8 (2C), 128.7, 128.6, 128.5, 128.3, 128.2, 128.1 (C.sub.Ar), 98.4 (C-1.sub.A-, .sup.1J.sub.C,H=168 Hz), 97.5 (C-1.sub.A-, .sup.1J.sub.C,H=169 Hz), 96.1 (C-1.sub.B-, .sup.1J.sub.C,H=163 Hz), 96.1 (C-1.sub.B-, .sup.1J.sub.C,H=176 Hz), 92.5, 92.2 (2C, CCl.sub.3), 76.5 (C-3.sub.B-), 75.2 (C-3.sub.B-), 75.1 (C-4.sub.A-), 75.1 (C-4.sub.A-), 73.8 (C-3.sub.A-), 73.1 (C-3.sub.A-), 72.8 (C-5.sub.A-), 72.2 (C-5.sub.A-), 71.9 (CH.sub.2Bn-), 71.8 (CH.sub.2Bn-), 69.7 (C-5.sub.B-), 67.8 (CH.sub.2Bn-6), 67.7 (CH.sub.2Bn-6), 65.4 (C-5.sub.B-), 65.0 (C-4.sub.B-), 64.5 (C-4.sub.B-), 58.4 (OCH.sub.3-), 58.0 (OCH.sub.3-), 55.7 (C-2.sub.A-), 53.6 (C-2.sub.A-), 53.5 (C-2.sub.B-), 50.9 (C-2.sub.B-), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.31H.sub.37Cl.sub.6N.sub.6O.sub.10, 863.0697; found 863.0700.

    [0629] (Benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl (N-phenyl)trifluoroacetimidate (39b) and 2-trichloromethyl [(benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-1,2,4,6-tetradeoxy--D-galactopyrano]-[2,1,d]-oxazoline (40b). Hemiacetal 38b (3.0 g, 3.55 mmol, 1.0 equiv.) was dissolved in acetone (40 mL). PTFACl (731 L, 4.61 mmol, 1.3 equiv.) was added followed by Cs.sub.2CO.sub.3 (1.27 g, 3.90 mmol, 1.1 equiv.). After stirring for 2 h at rt, the reaction mixture was filtered, washed with acetone (2*20 mL) and the filtrate was concentrated under reduced pressure. Flash chromatography of the crude material eluting with cHex/EtOAc/Et.sub.3N (20:1:0.2) gave a mixture of the (N-phenyl)trifluoroacetimidate donor 39b and a minor amount of oxazoline 40b (3.0 g, 2.95 mmol, 83% based on 39b) as a white solid. The disaccharide donor 39b had R.sub.f 0.7 (Tol/EtOAc 4:1); HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.39H.sub.41Cl.sub.6F.sub.3N.sub.7O.sub.10, 1034.0970; found 1034.0993. Oxazoline 40b R.sub.f 0.5 (Tol/EtOAc 4:1); HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.31H.sub.32Cl.sub.6N.sub.5O.sub.9, 828.0326; found 828.0305.

    [0630] Oligosaccharides Equipped with a 4.sub.A-Endchain Methyl Group: Chain Elongation

    ##STR00105##

    [0631] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (41b). PTFACl (110 L, 692 mol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (191 mg, 586 mol, 1.1 equiv.) were added to hemiacetal 38b (450 mg, 533 mol, 1.0 equiv.) in acetone (10 mL). After stirring for 2 h at rt, the reaction mixture was passed through pad of Celite and solids were washed with acetone. The filtrate was concentrated and dried under vacuum to give the crude mix of donors 39b/40b (540 mg, quant.).

    [0632] A mix of crude donors 39b/40b (540 mg, 533 mol, 1.0 equiv. theo.) and acceptor 7 (464 mg, 533 mol, 1.0 equiv.) was coevaporated with anhyd toluene (10 mL) and dried under vacuum for 1 h. 4 MS (1 g) was added to a solution of the later in anhyd. DCM (20 mL) and the suspension was stirred for 45 min under an Ar atmosphere at rt. After cooling to 10 C., TfOH (2.4 L, 27 mol, 0.05 equiv.) was added and stirring was continued for 30 min while the bath temperature was kept at 0 C. A TLC analysis (Tol/EtOAc 4:1) showed the absence of donor 39b/40b and the presence of a new spot. At completion, Et.sub.3N (5 L) was added. The suspension was filtered through a fitted funnel and solids were washed with DCM (10 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (cHex/EtOAc 85:15.fwdarw.75:25) to give tetrasaccharide 41b as a white solid (730 mg, 0.429 mmol, 80%). The coupling product 41b had R.sub.f 0.55 (Tol/EtOAc 7:3). .sup.1H NMR (CDCl.sub.3) 7.45-7.17 (m, 20H, H.sub.Ar), 6.98 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B), 6.77 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.64 (d, 1H, J.sub.2,NH=8.0 Hz, NH.sub.A), 6.49 (d, 1H, J.sub.2,NH=6.4 Hz, NH.sub.A), 5.89-5.80 (m, 1H, CH.sub.All), 5.33-5.14 (m.sub.po, 6H, CH.sub.2Bn-6, CH.sub.2All), 5.16 (d.sub.po, 1H, J.sub.1,2=5.6 Hz, H-1.sub.A), 5.07 (d.sub.po, 1H, J.sub.1,2=5.6 Hz, H-1.sub.A), 4.83 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.79 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.77 (d.sub.po, 1H, J.sub.4,5=5.2 Hz, H-5.sub.A), 4.71-4.64 (m, 4H, H-3.sub.B, H-3.sub.B1, H-4.sub.A, H-5.sub.A) 4.54 (d, 1H, CH.sub.2Bn), 4.45 (brs, 2H, CH.sub.2Bn), 4.33-4.28 (m, 1H, CH.sub.2All), 4.23 (brs, 1H, H-4.sub.A1) 4.21-4.16 (m, 1H, H-2.sub.A), 4.06-4.01 (m, 1H, CH.sub.2All), 3.92 (d, 1H, J.sub.3,4=3.2 Hz, H-4.sub.B), 3.90-3.84 (m, 4H, H-2.sub.A1, H-3.sub.A, H-3.sub.A1, H-4.sub.A1), 3.78 (d, 1H, J.sub.3,4=3.2 Hz, H-4.sub.B1), 3.62-3.57 (m, 2H, H-2.sub.B, H-5.sub.B), 3.50-3.43 (m, 2H, H-2.sub.B, H-5.sub.B), 3.39 (s, 3H, OCH.sub.3), 1.30 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B), 1.19 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.8, 168.2 (CO.sub.CO2Bn), 162.0, 161.9, 161.6 (CO.sub.NHTCA), 137.4, 137.1, 135.1, 135.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 129.1, 129.0, 128.9, 128.7 (2C), 128.6, 128.5, 128.4, 128.3, 128.2, 128.0, 125.2 (C.sub.Ar), 117.9 (CH.sub.2All), 99.0 (C-1.sub.A1, .sup.1J.sub.C,H=170.8 Hz), 98.9 (C-1.sub.B1, .sup.1J.sub.C,H=165.7 Hz), 98.1 (C-1.sub.A, .sup.1J.sub.C,H=170.4 Hz), 97.8 (C-1.sub.B, .sup.1J.sub.C,H=162.3 Hz), 92.4, 92.3 (2C), 92.2 (CCl.sub.3), 75.0 (3C), 74.7 (C-5.sub.A, C-4.sub.A, C-3.sub.B, C-3.sub.B1), 73.2, 73.0, (C-3.sub.A, C-3.sub.A1), 72.5 (CH.sub.2Bn), 72.0 (CH.sub.2Bn), 71.9 (C-4.sub.A1), 71.5 (C-5.sub.A1), 70.0 (CH.sub.2All), 69.3, 68.8 (C-5.sub.B, C-5.sub.B1), 67.6, 67.4 (CH.sub.2Bn-6), 65.5, 65.1 (C-4.sub.B, C-4.sub.B1), 58.1 (OCH.sub.3), 55.6, 55.5 (C-2.sub.B, C-2.sub.B1), 54.1, 52.9 (C-2.sub.A, C-2.sub.A1), 17.3, 17.2 (C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.34H.sub.70Cl.sub.12N.sub.11O.sub.19 1716.1106, found 1716.1147.

    [0633] (Benzyl 3-O-benzyl-4-O-methyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranose (42b). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (16 mg, 19 mol, 0.05 equiv.) was dissolved in anhyd. THF (3.0 mL) and stirred for 30 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly and transferred into a solution of allyl glycoside 41b (650 mg, 0.383 mmol, 1.0 equiv.) in anhyd. THF (12 mL). After stirring for 2 h at rt, NIS (103 mg, 0.459 mmol, 1.2 equiv.) was added. After stirring for another 2 h at rt, the reaction was quenched with 10% aq. sodium sulphite. Volatiles were evaporated and the aq. phase was extracted with DCM (315 mL). The organic phases were pooled, washed with brine, dried over anhyd. Na.sub.2SO.sub.4 and concentrated. Purification by flash chromatography (Tol/EtOAc 70:30.fwdarw.65:35) gave hemiacetal 42b (/ 5:1, 550 mg, 331 mol, 86%) as a white foam. The /-anomer had R.sub.f 0.35, 0.55 (Tol/EtOAc 7:3). The -anomer had .sup.1H NMR (CDCl.sub.3) 7.45-7.17 (m, 20H, H.sub.Ar), 7.01 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B), 6.77 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.64 (d, 1H, J.sub.2,NH=8.0 Hz, NH.sub.A), 6.49 (d, 1H, J.sub.2,NH=6.4 Hz, NH.sub.A), 5.33-5.18 (m, 4H, CH.sub.2Bn-6), 5.08 (d, 1H, J.sub.1,2=7.2 Hz, H-1.sub.A), 5.04 (d, 1H, J.sub.1,2=5.6 Hz, H-1.sub.A1), 4.87 (d.sub.po, 1H, J.sub.1,2=8.0 Hz, H-11), 4.85 (dd.sub.po, 1H, H-1.sub.B), 4.76 (d.sub.po, 1H, J.sub.4,5=5.0 Hz, H-5.sub.A), 4.71 (dd.sub.po, 1H, J.sub.2,3=10.8 Hz, J.sub.3,4=3.6 Hz, H-3.sub.B), 4.68-4.63 (m.sub.po, 3H, CH.sub.2Bn), 4.56 (d, 1H, J=12.0 Hz, CH.sub.2Bn), 4.48 (dd, 2H, J=12.4 Hz, CH.sub.2Bn), 4.23-4.17 (m, 2H, H-2.sub.A, H-4.sub.A), 3.92-3.81 (m, 5H, H-2.sub.A, H-3.sub.A, H-3.sub.A1, H-4.sub.B), 3.78 (brd, 1H, J.sub.3,4=3.6 Hz, H-4.sub.B1), 3.35-3.42 (m, 4H, H-2.sub.B, H-2.sub.B1, H-5.sub.B, H-5.sub.B1), 3.39 (s, 3H, OCH.sub.3), 1.22, 1.18 (2d, 6H, J.sub.5,6=6.0 Hz, H-6.sub.B1, H-6.sub.B1). .sup.13C NMR (CDCl.sub.3) 168.9, 168.2 (2C, C-6.sub.A, C-6.sub.A1), 162.3, 162.0, 161.6 (2C) (CO.sub.NHTCA), 137.4, 137.0, 135.1, 135.0 (C.sub.q,Ar), 129.1, 129.0, 128.7 (2C), 128.6, 128.4 (2C), 128.3, 128.2, 128.0, 125.2 (C.sub.Ar), 99.1, 98.8, 98.2, 96.7, 92.4, 92.3, (C-1), 92.2 (2C, CCl.sub.3), 77.3 (CH.sub.2Bn), 77.2, 75.0 (3C, C-5.sub.A, C-4.sub.A, C-3.sub.B), 74.4 (C-3.sub.B1), 73.1, 72.9 (2C, C-3.sub.A, C-3.sub.A1), 72.0 (CH.sub.2Bn), 71.9 (C-4.sub.A1), 71.4 (C-5.sub.A1), 69.2, 68.9 (2C, C-5.sub.B, C-5.sub.B1), 67.5, 67.4 (CH.sub.2Bn-6), 65.1 (2C, C-4.sub.B, C-4.sub.B1), 58.1 (OCH.sub.3), 55.5, 55.1 (2C, C-2.sub.B, C-2.sub.B1), 53.9, 52.8 (2C, C-2.sub.A, C-2.sub.A1), 17.4, 17.2 (2C, C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.61H.sub.66Cl.sub.12N.sub.11O.sub.19 1676.0793, found 1676.0797.

    [0634] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (43b). [2+4]-Glycosylation: PTFACl (73 L, 462 mol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (127 mg, 391 mol, 1.1 equiv.) were added to hemiacetal 38b (300 mg, 355 mol, 1.0 equiv.) in acetone (10 mL). After stirring for 2 h at rt, the reaction mixture was filtered through Celite bed and washed with acetone (5 mL) twice. The filtrate was concentrated under reduced pressure to give the crude donors 39b/40b (360 mg, 0.355 mmol, quant.), which was used as such in the next step after extensive drying under high vacuum.

    [0635] A mix of donors 39b/40b (355 mol, 1.0 equiv. theo.) and acceptor 15b (598 mg, 355 mol, 1.0 equiv.) were coevaporated with anhyd. toluene (10 mL) twice and then dried extensively under high vacuum. The mixture was stirred with freshly activated MS 4 (1.0 g) in anhyd. DCM (12 mL) for 45 min under an Ar atmosphere at rt. After cooling to 0 C., TMSOTf (3.2 L, 18 mol, 0.05 equiv.) was added and stirring was continued for 50 min while keeping the bath temperature at 0 C. A TLC analysis (Tol/EtOAc 6:4) showed the absence of donors 39b/40b and the presence of a new spot. At completion, Et.sub.3N (3 L) was added. The suspension was passed through a fitted funnel and solids were washed with DCM (5 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 82:18.fwdarw.85:15) to give hexasaccharide 43b as a white solid (700 mg, 278 mol, 78%). The coupling product 43b had R.sub.f 0.4 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.94H.sub.103Cl.sub.18N.sub.17O.sub.28 1273.5770; found 1273.5784.

    [0636] (Benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranose (44b). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (12 mg, 14 mol, 0.05 equiv.) was dissolved in anhyd. THF (4 mL) and the solution was stirred for 20 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly and poured into a solution of allyl glycoside 43b (700 mg, 278 mol, 1.0 equiv.) in anhyd. THF (6 mL). After stirring for 2 h at rt, NIS (94 mg, 418 mol, 1.5 equiv.) was added. After stirring for another 2 h at rt, the reaction was quenched with 10% aq sodium sulphite. Volatiles were eliminated under vacuum and the aq. phase was extracted repeatedly with DCM (10 mL). The organic layers were pooled, washed with brine, dried over anhyd. Na.sub.2SO.sub.4 and concentrated. Purification by flash chromatography (Tol/EtOAc 75:25.fwdarw.65:35) gave hemiacetal 44b (/ mixture, 540 mg, 219 mol, 78%) as a white foam, R.sub.f 0.25, 0.4 (Tol/EtOAc 7:3).

    [0637] Allyl (benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (45b). [2+6]-Glycosylation: A mix of the PTFA donor donors 39b/40b (200 mg, 0.197 mmol, 1.4 equiv.) and acceptor 17b (344 mg, 0.138 mmol, 1.0 equiv.) were stirred with freshly activated MS 4 (600 mg) in anhyd. DCM (6 mL) for 30 min under an Ar atmosphere at rt. After cooling to 10 C., TMSOTf (1.8 L, 10 mol, 0.05 equiv.) was added and stirring was continued for 1 h while keeping the bath temperature at 10 C. At completion, Et.sub.3N (3 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (6 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 85:15.fwdarw.82:18) to give octasaccharide 45b as a white solid (350 mg, 0.105 mmol, 76%). The coupling product 45b had R.sub.f 0.35 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.124H.sub.128Cl.sub.24N.sub.21O.sub.37 3356.1160; found 3356.1186.

    [0638] (Benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranose (46b). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (4.0 mg, 5 mol, 0.05 equiv.) was dissolved in anhyd THF (2.0 mL) and stirred for 40 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed with a flow of Ar and poured into a solution of octasaccharide 45b (700 mg, 0.105 mmol, 1.0 equiv.) in anhyd. THF (3.0 mL). After stirring for 2 h at rt, NIS (31 mg, 317 mol, 1.3 equiv.) was added. After stirring for another 2 h at rt, the reaction was quenched by adding 10% aq. sodium sulphite. The reaction mixture was concentrated and the aq. phase was extracted with DCM (5 mL) three times. The organic phases were pooled, washed with brine, dried over anhyd. Na.sub.2SO.sub.4 and concentrated. Purification by flash chromatography (Tol/EtOAc 70:30.fwdarw.60:40) gave the corresponding hemiacetal 46b (/ mixture, 280 mg, 85 mol, 81%) as a white foam. Hemiacetal 46b had R.sub.f 0.15, 0.25 (Tol/EtOAc 6:4). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.121H.sub.128Cl.sub.24N.sub.22O.sub.37 1662.0643; found 1662.0621.

    Example 8. Linker-Equipped Oligosaccharides Featuring a 4.SUB.A.-Me at the Endchain A Residue: Linker=(S)-()-2,3-dibenzyloxy-1-propanol

    [0639] Linker Attachment

    ##STR00106##

    [0640] (S)-2,3-Dibenzyloxy-1-propyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (47b). PTFACl (212 L, 1.33 mmol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (369 mg, 1.13 mmol, 1.1 equiv.) were added to hemiacetal 9a (1.0 g, 1.03 mmol, 1.0 equiv.) in acetone (20 mL). After stirring for 2 h at rt, the reaction mixture was filtered through a pad of Celite and solids were washed with acetone (10 mL) twice. The filtrate was concentrated and dried under vacuum to give the crude PTFA/oxazoline donors 9b/10b (600 mg, 0.592 mmol, quant.), which was used as such in the next step after extensive drying under vacuum.

    [0641] A mix of the crude donors 9b/10b (1.17 g, 1.03 mmol, 1.0 equiv. theo.) and (S)-()-2,3-dibenzyloxy-1-propanol (519 L, 2.06 mmol, 2.0 equiv.) was stirred with freshly activated MS 4.sub.A (1 g) in anhyd. DCM (20 mL) for 1 h under an Ar atmosphere at rt. After cooling to 10 C., TfOH (4.5 L, 51 mol, 0.05 equiv.) was added and stirring was continued for 30 min during which the bath temperature was kept at 10 C. A TLC analysis (Tol/EtOAc 4:1) showed the absence of donors 9b/10b and the presence of a new spot. At completion, Et.sub.3N (10 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (20 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (cHex/EtOAc 80:20.fwdarw.76:24) to give disaccharide 47b as a white solid (1.1 g, 0.897 mmol, 87%). The coupling product 47b had R.sub.f 0.6 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.85-7.74 (m, 3H, H.sub.Ar), 7.70 (brs, 1H, H.sub.Ar), 7.45-7.48 (m, 2H, H.sub.Ar), 7.45-7.48 (m, 21H, H.sub.Ar), 6.71 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.A), 6.65 (d, 1H, J.sub.2,NH=7.8 Hz, NH.sub.B), 5.23 (brs, 2H, CH.sub.2Bn), 5.18 (d, 1H, J.sub.1,2=5.2 Hz, H-1.sub.A), 4.86 (d, 1H, J.sub.4,5=5.2 Hz, H-5.sub.A), 4.73 (brs, 2H, CH.sub.2Bn), 4.73 (d.sub.po, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B), 4.66 (brs, 2H, CH.sub.2Bn), 4.58-4.54 (m, 3H, CH.sub.2Bn), 4.49-4.45 (m.sub.po, 3H, CH.sub.23n, H-3.sub.B), 4.22 (ddd.sub.po, 1H, H-2.sub.A), 4.08 (dd, 1H, J.sub.3,4=2.8 Hz, H-4.sub.A), 3.96 (dd, 1H, J=4.4 Hz, 10.4 Hz, H.sub.2-linker), 3.85-3.82 (m, 2H, H-3.sub.A, H-4.sub.B), 3.78-3.73 (m, 1H, H.sub.1-linker), 3.67-3.52 (m, 5H, H.sub.1-linker, H.sub.3-linker, H-2.sub.B, H-5.sub.B), 1.27 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.9 (C-6.sub.A), 161.9, 161.6 (CO.sub.NHTCA), 138.5, 138.3, 137.4, 135.0, 133.1 (2C) (C.sub.q,Ar), 128.7, 128.6, 128.4, 128.3 (2C), 128.2, 127.9 (2C), 127.7 (2C), 127.6, 127.5 (2C), 127.0, 126.2, 126.1, 125.9 (C.sub.Ar), 99.4 (C-1.sub.B, .sup.1J.sub.C,H=163.4 Hz), 98.9 (C-1.sub.A, .sup.1J.sub.C,H=171.0 Hz), 92.3, 92.1 (2C, CCl.sub.3), 77.0 (CH), 76.3 (C-3.sub.B), 73.4 (C-3.sub.A), 73.3 (CH.sub.2Bn), 72.1 (CH.sub.2Nap), 72.0 (2C, C-4.sub.A, CH.sub.2Bn), 71.9 (CH.sub.2Bn), 71.6 (C-5.sub.A), 70.1 (CH.sub.2Bn), 69.4 (C-5.sub.B), 68.8 (CH.sub.2Bn), 67.5 (CH.sub.2Bn-6), 65.1 (C-4.sub.B), 55.2 (C-2.sub.B), 53.0 (C-2.sub.A), 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.58H.sub.61Cl.sub.6N.sub.6O.sub.2 1243.2473; found 1243.2493.

    [0642] (S)-2,3-Dibenzyloxy-1-propyl (benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (48b). DDQ (482 mg, 2.12 mmol, 2.6 equiv.) was added to disaccharide 47b (1.0 g, 816 mol, 1.0 equiv.) in a mix of DCM (20.0 mL) and phosphate buffer pH 7 (2.0 mL). The biphasic mixture was cooled to 0 C. and stirred vigorously for 2 h. A TLC analysis (Tol/EtOAC 4:1) showed the absence of the fully protected 47b and the presence of a more polar spot. 10% Aq. NaHCO.sub.3 (20 mL) was added followed by DCM (20 mL). The DCM layer was separated, washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/EtOAc 80:20.fwdarw.75:25) to give alcohol 48b (585 mg, 539 mol, 66%) as a white solid. Disaccharide 48b had R.sub.f 0.5 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.43-7.26 (m, 20H, H.sub.Ar), 6.80 (d, 1H, J.sub.2,NH=8.2 Hz, NH.sub.A), 6.73 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 5.26 (dd.sub.po, 2H, J=12.0 Hz, CH.sub.2Bn), 4.96 (d, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A), 4.76 (d.sub.po, 1H, J.sub.1,2=8.2 Hz, H-1.sub.B), 4.75-4.68 (m.sub.po, 3H, H-5.sub.A, CH.sub.2Bn), 4.65 (dd.sub.po, 2H, CH.sub.2Bn), 4.53 (dd.sub.po, 2H, CH.sub.2Bn), 4.47 (dd, 1H, J.sub.2,3=10.6 Hz, J.sub.3,4=3.6 Hz, H-3.sub.B), 4.34-4.29 (m.sub.po, 1H, H-2.sub.A), 4.08 (dt, 1H, J.sub.4,5=7.6 Hz, J.sub.3,4=3.6 Hz, H-4.sub.A), 3.98 (dd, 1H, J=3.6, 5.6 Hz, H.sub.3-linker), 3.84 (dd, 1H, J=3.6, 5.6 Hz, H-3.sub.A), 3.77-3.72 (m, 2H, H-4.sub.B, H.sub.2-linker), 2.82 (d, 1H, J.sub.4,OH=8.0 Hz, OH), 3.68-3.55 (m, 5H, H.sub.1-linker, H.sub.3-linker, H-2.sub.B, H-5.sub.B), 1.27 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.8 (C-6.sub.A), 162.1, 161.6 (CO.sub.NHTCA), 138.4, 138.2, 137.1, 134.9 (C.sub.q,Ar), 128.7, 128.5, 128.4 (2C), 128.3 (2C), 128.2, 127.6 (2C) (C.sub.Ar), 99.5 (C-1.sub.A, .sup.1J.sub.C,H=171.2 Hz), 99.3 (C-1.sub.B, .sup.1J.sub.C,H=163.4 Hz), 92.2, 91.9 (2C, CCl.sub.3), 77.0 (CH), 76.7 (C-3.sub.B), 74.7 (C-3.sub.A), 73.3 (CH.sub.2Bn), 72.2 (CH.sub.2Bn), 72.1 (CH.sub.2Bn), 71.2 (C-5.sub.A), 70.0 (CH.sub.2Bn), 69.6 (C-5.sub.B), 68.8 (CH.sub.2Bn), 67.5 (CH.sub.2Bn-6), 65.5 (C-4.sub.A), 65.3 (C-4.sub.B), 55.4 (C-2.sub.B), 51.3 (C-2.sub.A), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.47H.sub.53Cl.sub.6N.sub.6O.sub.12, 1103.1858; found 1103.1847.

    [0643] (S)-2,3-Dibenzyloxy-1-propyl (benzyl 3-O-benzyl-2-deoxy-4-O-methyl-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (49b). PTFACl (122 L, 770 mol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (212 mg, 651 mol, 1.1 equiv.) were added to hemiacetal 38b (500 mg, 592 mol, 1.0 equiv.) in acetone (15 mL). After stirring for 2 h at rt, the reaction mixture was filtered through a pad of Celite bed and solids were washed with acetone. Volatiles were evaporated to give a mix of the crude donors 39b/40b (600 mg, quant.), which was used as such in the next step after extensive drying under vacuum.

    [0644] A mix of the crude donors 39b/40b (600 mg, 592 mol, 1.0 equiv. theo.) and (S)-()-2,3-dibenzyloxy-1-propanol (298 L, 1.18 mmol, 2.0 equiv.) was stirred with freshly activated 4.sub.A MS (500 mg) in anhyd. DCM (20 mL) for 1 h under an Ar atmosphere at rt. After cooling to 0 C., TfOH (2.6 L, 30 mol, 0.05 equiv.) was added and stirring was continued at this temperature for 40 min. A TLC analysis (Tol/EtOAc 4:1) showed the absence of donors 39b/40b and the presence of a new spot. At completion, Et.sub.3N (5 L) was added. The suspension was passed through a fitted funnel and solids were washed with DCM (10 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (cHex/EtOAc 80:20.fwdarw.76:24) to give disaccharide 49b as a white solid (430 mg, 423 mol, 66%). The coupling product 49b had R.sub.f 0.8 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.42-7.26 (m, 20H, H.sub.Ar), 6.74 (dd.sub.po, 2H, J.sub.2,NH=7.6 Hz, NH.sub.A, NH.sub.B), 5.26 (brd, 2H, CH.sub.2Bn-6), 5.20 (d, 1H, J.sub.1,2=5.6 Hz, H-1.sub.A), 4.79 (d, 1H, J.sub.4,5=4.8 Hz, H-5.sub.A), 4.73 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.66-4.63 (m, 3H, CH.sub.2Bn), 4.57-4.53 (m, 3H, CH.sub.2Bn), 4.49 (dd, 1H, J.sub.2,3=10.8 Hz, J.sub.3,4=3.6 Hz, H-3.sub.B), 4.15-4.10 (m, 1H, H-2.sub.A), 3.97-3.92 (m, 2H, H-3.sub.A, OCH.sub.2), 3.86-3.82 (m, 2H, H-4.sub.B, H-4.sub.A), 3.76-3.72 (m, 1H, CH), 3.67-3.54 (m, 5H, H-2.sub.B, H-5.sub.B, 3OCH.sub.2), 3.39 (s, 3H, OCH.sub.3), 1.26 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.9 (C-6.sub.A), 161.9, 161.7 (CO.sub.NHTCA), 138.5, 138.3, 137.4, 135.0 (C.sub.q,Ar), 128.8, 128.7 (2C), 128.4, 128.3 (2C), 128.2, 128.0, 127.6, 127.5 (3C) (C.sub.Ar), 99.4 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 98.6 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 92.3, 92.2 (2C, CCl.sub.3), 77.0 (CH), 76.2 (C-3.sub.B), 75.1 (C-4.sub.A), 73.3 (CH.sub.2Bn), 73.1 (C-3.sub.A), 72.1 (CH.sub.2Bn), 72.0 (CH.sub.2Bn), 71.4 (C-5.sub.A), 70.1 (OCH.sub.2), 69.4 (C-5.sub.B), 68.8 (OCH.sub.2), 67.5 (CH.sub.2Bn-6), 65.1 (C-4.sub.B), 58.0 (OCH.sub.3), 55.2 (C-2.sub.B), 53.2 (C-2.sub.A), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.48H.sub.55Cl.sub.6N.sub.6O.sub.12, 1117.2004; found 1117.1999.

    [0645] (S)-2,3-Dibenzyloxy-1-propyl (benzyl 3-O-benzyl-4-O-methyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (50b). PTFA-Cl (73 L, 0.460 mmol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (127 mg, 385 mol, 1.1 equiv.) were added to hemiacetal 38b (300 mg, 353 mol, 1.0 equiv.) in acetone (10 mL). After stirring for 2 h at rt, the reaction mixture was filtered through a pad of Celite and washed with acetone (5 mL) twice. The filtrate was concentrated under reduced pressure to give the crude PTFA donor (360 mg, quant.), which was used as such in the next step after extensive drying under high vacuum.

    [0646] A mix of the crude PTFA donor (360 mg, 0.355 mmol, 1.25 equiv. theo.) and acceptor 48b (308 mg, 284 mol, 1.0 equiv.) were coevaporated with anhyd. toluene (5 mL) twice and then dried extensively under high vacuum. Freshly activated 4 MS (600 mg) was added to the mixture in anhyd. DCM (9.0 mL) and the suspension was stirred for 1 h under an Ar atmosphere at rt. After cooling to 10 C., TfOH (1.6 L, 18 mol, 0.06 equiv.) was added and stirring was continued for 30 min while keeping the bath temperature at 0 C. A TLC analysis (Tol/EtOAc 14:6) showed the absence of donor and the presence of a new spot. At completion, Et.sub.3N (3 L) was added. The suspension was passed through a fitted funnel and solids were washed with DCM (5 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (cHex/EtOAc 82:18.fwdarw.77:23) to give tetrasaccharide 50b as a white solid (490 mg, 256 mol, 90%). The coupling product 50b had R.sub.f 0.7 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.78H.sub.84Cl.sub.12N.sub.11O.sub.21 1930.2100; found 1930.2138.

    [0647] (S)-2,3-Dibenzyloxy-1-propyl (benzyl 3-O-benzyl-4-O-methyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-4-O-methyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (51b). Hemiacetal 44b (540 mg, 219 mol) was dissolved in acetone (6 mL). PTFA-Cl (57 L, 357 mol, 1.6 equiv.) and Cs.sub.2CO.sub.3 (102 mg, 0.313 mmol, 1.4 equiv.) were added. After stirring for 2 h at rt, the suspension was passed through a pad of Celite and solids were washed with acetone (5 mL) twice. Volatiles were evaporated to give the crude PTFA donor (577 mg, quant.), which was used as such in the next step after extensive drying under vacuum.

    [0648] A mix of the crude donor (577 mg, 219 mol, 1.0 equiv. theo.) and (S)-()-2,3-dibenzyloxy-1-propanol (169 L, 670 mol, 3.0 equiv.) in anhyd. DCM (12 mL) containing freshly activated MS 4 (1.2 g) was stirred for 30 min under an Ar atmosphere at rt. After cooling to 10 C., TMSOTf (2.0 L, 11 mol, 0.05 equiv.) was added and stirring was continued for 40 min while keeping the bath temperature at 10 C. At completion, Et.sub.3N (4 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (6 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 82:18.fwdarw.85:15) to give hexasaccharide 51b as a white solid (380 mg, 139 mol, 64%). The coupling product 51b had R.sub.f 0.45 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.108H.sub.118Cl.sub.18N.sub.15O.sub.30 2243.2255; found 2746.2190.

    [0649] (S)-2,3-Dibenzyloxy-1-propyl (benzyl 3-O-benzyl-4-O-methyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-4-O-methyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-4-O-methyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (52b). PTFACl (18 L, 111 mol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (31 mg, 94 mol, 1.1 equiv.) were added to a solution of hemiacetal 46b (280 mg, 85 mol, 1.0 equiv.) in acetone (5.0 mL). After stirring for 2 h at rt, the reaction mixture was passed through a pad of Celite and solids were washed with acetone (3 mL) twice. The filtrate was concentrated under reduced pressure to give the crude donor (300 mg, quant.), which was used as such in the next step after extensive drying under vacuum.

    [0650] A mix of the crude PTFA donor (300 mg, 85 mol, 1.0 equiv. theo.) and (S)-()-2,3-dibenzyloxy-1-propanol (85 L, 341 mol, 4.0 equiv.) in anhyd. DCM (6.0 mL) containing freshly activated MS 4 (300 mg) was stirred for 30 min under an Ar atmosphere at rt. After cooling to 10 C., TMSOTf (1.0 L, 4 mol, 0.05 equiv.) was added and stirring was continued for 45 min while keeping the temperature of the bath at 10 C. At completion, Et.sub.3N (2 L) was added. After stirring at this temperature for another 10 min, the suspension was passed through a fitted funnel and solids were washed with DCM (4 mL) twice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 84:16.fwdarw.78:22) to give octasaccharide 52b as a white solid (155 mg, 44 mmol, 51%). The condensation product 52b had R.sub.f 0.6 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.138H.sub.146Cl.sub.24N.sub.22O.sub.39 1793.1260; found 1793.1251.

    [0651] Full Deprotection

    ##STR00107##

    [0652] (S)-2,3-Dihydroxy-1-propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (53b). The fully protected disaccharide 49b (120 mg, 109 mol) was subjected to hydrogenation-mediated deprotection (protocol 1). Disaccharide 53b was obtained as a white solid (48 mg, 94 mol, 86%). Disaccharide 53b had RP-HPLC (215 nm) R.sub.t=12.3 min (conditions A), R.sub.t=6.3 min (conditions B). .sup.1H NMR (D.sub.2O) 4.88 (d, 1H, J.sub.1,2=8.3 Hz, H-1.sub.A), 4.78 (brs, 1H, H-5.sub.A), 4.48 (d, 1H, J.sub.1,2=8.6 Hz, H-1.sub.B), 4.13 (dd, 1H, J.sub.3,4=4.4 Hz, J.sub.2,3=10.8 Hz, H-3.sub.B), 4.02 (q.sub.po, 1H, H-5.sub.B), 3.98 brt, 1H, H-4.sub.A), 3.90 (dd.sub.po, 1H, H-2.sub.A), 3.85 (brd, 1H, H-4.sub.B), 3.84-3.74 (m.sub.po, 3H, H-2.sub.B, H.sub.1-linker), 3.70 (dd, 1H, J.sub.2,3=10.8 Hz, J.sub.3,4=2.0 Hz, H-3.sub.A), 3.62-3.55 (m, 2H, H.sub.1-linker), 3.49-3.45 (m.sub.po, 1H, H.sub.2-linker), 3.48 (brs, 3H, OCH.sub.3), 2.00, 1.96 (2s, 6H, CH.sub.3Ac), 1.29 (d, 3H, J.sub.5,6=6.6 Hz, H-6.sub.B). .sup.13C NMR (D.sub.2O) 174.5, 174.2 (NHCO.sub.A,B), 172.7 (C-6.sub.A), 101.7 (C-1.sub.B, .sup.1J.sub.C,H=163.0 Hz), 100.8 (C-1.sub.A, .sup.1J.sub.C,H=169.0 Hz), 78.6 (C-4.sub.A), 76.1 (C-3.sub.B), 72.8 (C-5.sub.A), 70.8 (OCH.sub.2), 70.2 (OCH.sub.linker), 67.6 (C-3.sub.A), 67.3 (C-5.sub.B), 62.3 (OCH.sub.2), 58.1 (OCH.sub.3), 54.7 (C-4.sub.B), 51.7 (C-2.sub.A), 50.8 (C-2.sub.B), 22.2 (2C, CH.sub.3Ac), 15.5 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.20H.sub.36N.sub.3O.sub.12, 510.2293; found 510.2292.

    [0653] (S)-2,3-Dihydroxy-1-propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (54b). The fully protected tetrasaccharide 50b (78 mg, 41 mol) was subjected to hydrogenation-mediated deprotection (protocol 1). Tetrasaccharide 54b was obtained as a white solid (13 mg, 14 mol, 35%). Tetrasaccharide 54b had RP-HPLC (215 nm) R.sub.t=7.9 min (conditions A). .sup.1H NMR (D.sub.2O, partial) 4.87 (d, 1H, J.sub.1,2=8.1 Hz, H-1.sub.A), 4.80-4.74 (m.sub.po, 4H, H-5.sub.A, H-5.sub.B1, H-1.sub.A, H-1.sub.B1), 4.48-4.45 (m, 2H, H-4.sub.A, H-1.sub.B), 4.16-4.10 (m, 2H, H-3.sub.B, H-3.sub.B1), 4.02-3.98 (m, 3H, H-5.sub.B, H-5.sub.B1), 3.93-3.89 (m.sub.po, 1H, H-2.sub.A), 3.86-3.66 (m, 9H), 3.59-3.55 (m, 2H, H.sub.1-linker), 3.49-3.47 (m.sub.po, 1H, H.sub.2-linker), 3.48 (brs, 3H, OCH.sub.3), 1.99, 1.97, 1.94 (2s, 12H, CH.sub.3Ac), 1.29 (d.sub.po, 6H, J.sub.5,6=6.0 Hz, H-6.sub.B, H-6.sub.B1). .sup.13C NMR (D.sub.2O, partial) 174.6, 174.5 (5C, 4NHCO, C-6.sub.A), 174.1 (C-6.sub.A), 102.8 (C-1.sub.B1, .sup.1J.sub.C,H=166.0 Hz), 101.8 (C-1.sub.B, .sup.1J.sub.C,H=162.4 Hz), 101.1 (C-1.sub.A, .sup.1J.sub.C,H=166.4 Hz), 100.9 (C-1.sub.A, .sup.1J.sub.C,H=167.0 Hz), 78.5 (C-4.sub.A), 76.5 (C-3.sub.B1), 76.0 (3C, C-3.sub.B, C-4.sub.A1, C-5.sub.A1), 72.7 (C-5.sub.A), 70.8 (OCH.sub.2), 70.2 (OCH.sub.linker), 67.6 (C-3.sub.A), 67.3 (2C, C-5.sub.B, C-5.sub.B1), 62.3, 58.2 (OCH.sub.2), 58.2 (OCH.sub.3), 54.8, 54.7 (2C, C-4.sub.B1, C-4.sub.B), 51.7, 51.5 (2C, C-2.sub.A, C-2.sub.A1), 51.0, 50.9 (2C, C-2.sub.B, C-2.sub.B1), 22.4, 22.2 (2C, CH.sub.3Ac), 15.6, 15.5 (2C, C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.36H.sub.61N.sub.6O.sub.21 913.3884; found 913.3864. HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.36H.sub.60N.sub.6O.sub.21Na, 935.3704; found 935.3680. HRMS (ESI.sup.+): m/z [M+2H]2.sup.+ calcd for C.sub.36H.sub.61N.sub.6O.sub.21 457.1979; found 457.1972.

    [0654] (S)-2,3-Dihydroxy-1-propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (55b) The fully protected hexasaccharide 51b (75 mg, 28 mol) was subjected to hydrogenation-mediated deprotection (protocol 1). Hexasaccharide 55b was obtained as a white solid (8.5 mg, 9.3 mol, 24%). .sup.1H NMR (D.sub.2O, partial) 4.90 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A), 4.81-4.70 (m.sub.po, 5H, H-5.sub.A, 2H-1.sub.A, 2H-1.sub.B), 4.70-4.65 (m.sub.po, 2H, 2H-5.sub.A), 4.49-4.48 (m, 3H, 2H-4.sub.A, H-1.sub.B), 4.17-4.11 (m, 3H, H-3.sub.B), 4.02-3.93 (m, 4H, H-4.sub.A, 3H-5.sub.B), 3.94-3.67 (m, 14H), 3.60-3.55 (m, 2H, H.sub.1-linker), 3.49 (brs, 3H, OCH.sub.3), 3.47-3.45 (m, 1H, H.sub.1-linker), 3.01-1.95 (m.sub.po, 18H, CH.sub.3Ac), 1.30 (d.sub.po, 9H, J.sub.5,6=6.0 Hz, H-6.sub.B), .sup.13C NMR (D.sub.2O, partial) 174.6, 174.5, 174.4, 174.1 (5C, 4NHCO.sub.A,B, C-6.sub.A2), 172.9, 172.4 (2C, C-6.sub.A, C-6.sub.A1), 102.9 (2C, C-1.sub.B1, C-1.sub.B2, .sup.1J.sub.C,H=166.0 Hz), 101.8 (C-1.sub.B, .sup.1J.sub.C,H=163.0 Hz), 101.1 (2C, C-1.sub.A, C-1.sub.A1, .sup.1J.sub.C,H=167.0 Hz), 100.9 (C-1.sub.A2, .sup.1J.sub.C,H=169.0 Hz), 78.6, 76.7 (C-4.sub.A, C-4.sub.A1), 76.3, 76.0 (C-5.sub.A, C-5.sub.A1), 75.9, 75.7 (C-3.sub.B), 72.9, 70.8 (2C, OCH.sub.2), 70.2 (OCH.sub.linker), 67.6 (3C, C-3.sub.A, C-3.sub.A1, C-3.sub.A2), 67.3 (3C, C-5.sub.B, C-5.sub.B1, C-5.sub.B2), 62.3 (OCH.sub.2), 58.1 (OCH.sub.3), 54.8 (3C, C-4.sub.B1, C-4.sub.B, C-4.sub.B2), 51.7 (3C, C-2.sub.A, C-2.sub.A1, C-2.sub.A2), 51.0, 50.9 (3C, C-2.sub.B, C-2.sub.B1, C-2.sub.B2), 22.3, 22.2 (6C, CH.sub.3Ac), 15.6, 15.5 (3C, C-6.sub.B, C-6.sub.B1, C-6.sub.B2). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.52H.sub.87N.sub.9O.sub.30 658.7774; found 658.7754.

    Example 9: Linker-Equipped Oligosaccharides Featuring a 4.SUB.A.-Endchain Hydroxyl Group

    [0655] Azidopropyl Aglycon as Linker Precursor

    ##STR00108##

    [0656] Scheme 22. Synthesis of azidopropyl-equipped AB oligomers. (i) 3-Azidopropanol, TMSOTf, DCE, 15 C., 89% for 56, 93% for 58b, (ii) DDQ, 10:1 DCM/Phosphate buffer pH 7, 86%, (iii) 9b/10b, TMSOTf, DCE, 15 C., 66%, (iv) 13b/14b, TMSOTf, DCE, 5 C., 55%.

    [0657] 3-Azidopropyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (56b) and 2-Azidopropyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (57b). A solution of donors 9b/10b (1.0 g, 876 mol, 1.0 equiv) and 3-azidopropanol (161 L, 1.75 mmol, 2.0 equiv) in anhyd. DCE (14 mL) was stirred with freshly activated 4 MS (1.0 g) for 30 min at rt under an Ar atmosphere. The reaction mixture was cooled to 15 C. and TMSOTf (11 L, 61 mol, 0.07 equiv.) was added. After 40 min at 15 C., the mixture was quenched with Et.sub.3N (12 L, 0.1 equiv.), filtered and concentrated. Flash chromatography (Tol/EtOAc 4:1) gave by order of elution the -isomer 57b (40 mg, 38 mol, 4%) as a white solid and the desired disaccharide 56b (830 mg, 787 mol, 89%) as a white solid. Disaccharide 56b had R.sub.f 0.6 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.85-7.74 (m, 3H, H.sub.Ar), 7.70 (brs, 1H, H.sub.Ar), 7.52-7.48 (m, 2H, H.sub.Ar), 7.42-7.27 (m, 11H, H.sub.Ar), 6.80 (d, 1H, J.sub.2,NH=7.4 Hz, NH.sub.B), 6.72 (d, 1H, J.sub.2,NH=7.9 Hz, NH.sub.A), 5.26-5.19 (dd.sub.po, 3H, H-1.sub.A, CH.sub.2Bn-6), 4.85 (d, 1H, J.sub.4,5=4.6 Hz, H-5.sub.A), 4.74 (brs, 2H, CH.sub.2Nap), 4.71 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.53 (d.sub.po, 1H, J=12.0 Hz, CH.sub.2Bn), 4.50 (dd.sub.po, 1H, J.sub.3,4=4.0 Hz, J.sub.2,3=10.2 Hz, H-3.sub.B), 4.45 (d.sub.po, 1H, CH.sub.2Bn), 4.24-4.19 (m, 1H, H-2.sub.A), 4.10 (dd, 1H, J.sub.3,4=2.8 Hz, H-4.sub.A), 3.95-3.89 (m, 1H, OCH.sub.2), 3.86 (brd, 1H, H-4.sub.B), 3.84 (dd, 1H, J.sub.2,3=8.1 Hz, H-3.sub.A), 3.67-3.51 (m.sub.po, H-2.sub.B, H-5.sub.B, OCH.sub.2), 3.37 (t, 2H, J=6.6 Hz, CH.sub.2N.sub.3), 1.89-1.77 (m, 2H, CH.sub.2), 1.28 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.9 (C-6.sub.A), 162.0, 161.7 (CO.sub.NHTCA), 137.4, 134.9, 134.5, 133.1 (2C) (C.sub.q,Ar), 128.7, 128.6, 128.4, 128.3, 128.2, 128.0, 127.9, 127.7, 127.0, 126.2, 126.1, 125.9 (C.sub.Ar), 99.1 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 98.8 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 92.4, 92.1 (2C, CCl.sub.3), 76.1 (C-3.sub.B), 73.4 (C-3.sub.A), 72.0 (C-4.sub.A, CH.sub.2Nap), 71.9 (CH.sub.2Bn), 71.8 (C-5.sub.A), 69.4 (C-5.sub.B), 67.5 (CH.sub.2Bn-6), 66.3 (OCH.sub.2), 65.1 (C-4.sub.B), 55.1 (C-2.sub.B), 53.1 (C-2.sub.A), 48.1 (CH.sub.2N.sub.3), 29.0, 17.3 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.44H.sub.48Cl.sub.6N.sub.9O.sub.10, 1072.1650, found 1072.1645.

    [0658] The side-product 57b had R.sub.f 0.65 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.86-7.75 (m, 3H, H.sub.Ar), 7.71 (brs, 1H, H.sub.Ar), 7.53-7.48 (m, 2H, H.sub.Ar), 7.45-7.16 (m, 11H, H.sub.Ar), 7.05 (d, 1H, J.sub.2,NH=7.4 Hz, NH.sub.B), 6.81 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.A), 5.41 (d, 1H, J.sub.1,2=6.2 Hz, H-1.sub.A), 5.19 (dd.sub.po, 2H, J=12.2 Hz, CH.sub.2Bn-6), 4.90 (d, 1H, J.sub.1,2=3.8 Hz, H-1.sub.B), 4.81 (d, 1H, J.sub.4,5=3.8 Hz, H-5.sub.A), 4.76 (brs, 2H, CH.sub.2Nap), 4.46-4.38 (m.sub.po, 3H, H-2.sub.B, CH.sub.2Bn), 4.24-4.15 (m.sub.po, 3H, H-2.sub.A, H-4.sub.A, H-3.sub.B), 3.94 (dd.sub.po, 1H, H-4.sub.B), 3.90-3.85 (m.sub.po, 2H, H-3.sub.A, H-5.sub.B), 3.76-3.70 (m, 1H, OCH.sub.2), 3.50-3.44 (m, 1H, OCH.sub.2), 3.35 (t, 2H, J=6.4 Hz, CH.sub.2N.sub.3), 1.85-1.78 (m, 2H, CH.sub.2), 1.24 (d, 3H, J.sub.5,6=6.2 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.9 (C-6.sub.A), 162.0, 161.7 (CO.sub.NHTCA), 136.9, 134.8, 134.5, 133.1 (2C) (C.sub.q,Ar), 129.0, 128.9, 128.8, 128.7, 128.6, 128.5, 128.4, 128.3, 128.2 (2C), 128.0, 127.9, 127.7, 127.1, 126.2, 126.1, 126.0, 125.2 (C.sub.Ar), 97.9 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 96.8 (C-1.sub.B, .sup.1J.sub.C,H=175 Hz), 92.5, 92.1 (2C, CCl.sub.3), 75.4 (C-3.sub.B), 73.1 (C-3.sub.A), 72.7 (C-5.sub.A), 72.0 (CH.sub.2Nap), 71.9 (C-4.sub.A), 71.6 (CH.sub.2Bn), 67.5 (CH.sub.2Bn-6), 65.6 (C-5.sub.B), 65.1 (OCH.sub.2), 64.9 (C-4.sub.B), 53.6 (C-2.sub.A), 50.8 (C-2.sub.B), 48.2 (CH.sub.2N.sub.3), 28.7 (CH.sub.2,linker), 17.2 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.44H.sub.48Cl.sub.6N.sub.9O.sub.10, 1072.1650, found 1072.1646.

    [0659] 3-Azidopropyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (58b). A solution of donor 13b/14b (3.8 g, 1.94 mmol, 1.0 equiv) and 3-azidopropanol (358 L, 3.88 mmol, 2.0 equiv) in anhyd DCE (50 mL) was stirred with freshly activated 4 MS (3.0 g) for 45 min at rt under an Ar atmosphere. The reaction mixture was cooled to 15 C. and TMSOTf (25 L, 136 mol, 0.07 equiv.) was added. After stirring for 1 h at 15 C., Et.sub.3N (20 L, 0.1 equiv.) was added. The suspension was passed through a pad of Celite, solids were washed with DCM, and volatiles were evaporated. Flash chromatography (Tol/ACN 90:10) of the residue gave the desired tetrasaccharide 58b (3.4 g, 1.82 mmol, 93%) as a white solid. Tetrasaccharide 58b had R.sub.f 0.55 (Tol/ACN 4:1). .sup.1H NMR (CDCl.sub.3) 7.84-7.74 (m, 3H, H.sub.Ar), 7.69 (brs, 1H, H.sub.Ar), 7.51-7.20 (m, 18H, H.sub.Ar), 7.08 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B1), 6.82 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.67 (d, 1H, J.sub.2,NH=8.0 Hz, NH.sub.A), 6.54 (d, 1H, J.sub.2,NH=7.6 Hz, NH.sub.A1), 5.30-5.18 (dd.sub.po, 4H, CH.sub.2Bn-6), 5.17 (d.sub.po, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A), 5.07 (d, 1H, J.sub.1,2=5.0 Hz, H-1.sub.A1), 4.83 (d.sub.po, 1H, J.sub.4,5=5.2 Hz, H-5.sub.A1), 4.81 (d.sub.po, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B1), 4.77-4.67 (m.sub.po, 4H, CH.sub.2Nap, H-5.sub.A, H-1.sub.B), 4.65 (dd.sub.po, 1H, J.sub.3,4=3.6 Hz, J.sub.2,3=10.6 Hz, H-3.sub.B1), 4.59-4.55 (m, 2H, H-3.sub.B, CH.sub.2Bn), 4.55 (d.sub.po, 1H, J=12.6 Hz, CH.sub.2Bn), 4.52 (brs, 2H, CH.sub.2Bn), 4.31-4.25 (m, 2H, H-4.sub.A, H-2.sub.B1), 4.08 (dd, 1H, J.sub.3,4=3.0 Hz, H-4.sub.A1), 3.94-3.88 (m.sub.po, 3H, H-2.sub.A, H-4.sub.B, OCH.sub.2), 3.84-3.73 (m.sub.po, H-3.sub.A, H-3.sub.A1, H-4.sub.A1), 3.68-3.60 (m, 2H, H-2.sub.B, H-5.sub.B), 3.56-3.51 (m, 2H, H-2.sub.B1, OCH.sub.2) 3.49-3.43 (m, 1H, H-5.sub.B1), 3.36 (t, 2H, J=6.6 Hz, CH.sub.2N.sub.3), 1.86-1.76 (m, 2H, CH.sub.2), 1.31 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B), 1.31 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B1). .sup.13C NMR (CDCl.sub.3) 168.9, 168.2 (2C, C-6.sub.A, C-6.sub.A1), 162.1, 161.9, 161.7, 161.6 (4C, CO.sub.NHTCA), 137.3, 137.0, 135.0, 134.9, 134.6, 133.1 (2C) (C.sub.q,Ar), 129.1, 129.0, 128.8 (2C), 128.7 (2C), 128.5, 128.4 (2C), 128.3, 128.2, 128.0, 127.9, 126.9, 126.2, 126.1, 125.8 (C.sub.Ar), 99.2 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz, C-1.sub.B1, .sup.1J.sub.C,H=166 Hz), 97.9 (C-1.sub.A, .sup.1J.sub.C,H=170.0 Hz), 92.4, 92.3, 92.1 (4C, CCl.sub.3), 75.2 (2C, C-5.sub.A, C-3.sub.B1) 74.8 (C-3.sub.B), 73.3, 73.1 (C-3.sub.A, C-3.sub.A1), 72.5 (CH.sub.2Nap), 72.0 (C-4.sub.A), 71.9 (CH.sub.2Bn), 71.8 (CH.sub.2Bn), 71.6 (C-4.sub.A1), 69.4 (C-5.sub.B), 68.9 (C-5.sub.B1), 67.6 (CH.sub.2Bn-6), 67.4 (CH.sub.2Bn-6), 66.2 (OCH.sub.2), 65.4, 65.1 (2C, C-4.sub.B, C-4.sub.B1), 55.4, 55.2 (2C, C-2.sub.B, C-2.sub.B1), 53.9, 52.7 (2C, C-2.sub.A, C-2.sub.A1), 48.1 (CH.sub.2N.sub.3), 29.0 (CH.sub.2,linker), 17.3, 17.2 (2C, C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.74H.sub.77Cl.sub.12N.sub.14O.sub.19 1885.1746, found 1885.1760.

    [0660] 3-Azidopropyl (benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (59b). Tetrasaccharide 58b (2.5 g, 1.33 mmol, 1.0 equiv.) was dissolved in DCM (25 mL) and phosphate buffer pH 7 (3 mL) was added. The biphasic mixture was cooled to 0 C. and DDQ (608 mg, 2.67 mmol, 2.0 equiv.) was added, and the biphasic mixture was stirred while keeping the bath temperature between 0-10 C. At completion, 5% aq. NaHCO.sub.3 (50 mL) was added and the biphasic mixture was diluted with DCM (50 mL). The DCM layer was separated, washed with brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (Tol/ACN 85:15.fwdarw.86:14) to give alcohol 59b (2.0 g, 1.15 mmol, 86%) as a white solid. Tetrasaccharide 59b had R.sub.f 0.35 (Tol/ACN 4:1). .sup.1H NMR (CDCl.sub.3) 7.84-7.74 (m, 20H, H.sub.Ar), 6.98 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B1), 6.79 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.75 (d, 1H, J.sub.2,NH=8.0 Hz, NH.sub.A), 6.52 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.A1), 5.31-5.20 (dd.sub.po, 4H, CH.sub.2Bn-6), 5.13 (d, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A), 4.90 (d, 1H, J.sub.1,2=4.0 Hz, H-1.sub.A1), 4.82 (d.sub.po, 1H, J.sub.4,5=8.4 Hz, H-1.sub.B), 4.72-4.69 (m.sub.po, 5H, CH.sub.2Bn, H-5.sub.A, H-5.sub.A1, H-1.sub.B1), 4.46-4.57 (m.sub.po, 2H, H-3.sub.B1, H-3.sub.B), 4.49-4.41 (m, 2H, CH.sub.2Bn), 4.34-4.30 (m, 1H, H-2.sub.A), 4.27 (t, 1H, J.sub.4,5=J.sub.3,4=2.4 Hz, H-4.sub.A), 4.15 (m.sub.po, 1H, H-4.sub.A1), 3.95-3.88 (m.sub.po, 3H, H-2.sub.A, H-4.sub.B, OCH.sub.2), 3.85 (d, 1H, J.sub.3,4=2.4 Hz, H-4.sub.A1), 3.82-3.78 (dd.sub.po, 1H, H-3.sub.A1), 3.73 (d, 1H, J.sub.3,4=3.0 Hz, H-4.sub.A), 3.65-3.42 (m.sub.po, 5H, H-2.sub.B, H-5.sub.B, H-2.sub.B1, OCH.sub.2, H-5.sub.B1), 3.35 (t, 2H, J=6.6 Hz, CH.sub.2N.sub.3), 2.83 (brs, 1H, OH), 1.88-1.76 (m, 2H, CH.sub.2), 1.30 (d, 3H, J.sub.5,6=6.0 Hz, H-6.sub.B), 1.17 (d, 3H, J.sub.5,6=6.2 Hz, H-6.sub.B1). .sup.13C NMR (CDCl.sub.3) 168.8, 168.2 (CO.sub.COOBn), 162.2, 161.9, 161.7, 161.6 (CO.sub.NHTCA), 137.8, 137.0, 136.9, 135.0, 134.9 (2C) (C.sub.q,Ar), 129.1, 129.0, 128.8, 128.7 (2C), 128.6, 128.5, 128.4 (2C), 128.2, 125.2 (C.sub.Ar), 99.5 (C-1.sub.A1, .sup.1J.sub.C,H=168 Hz), 99.0 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 99.0 (C-1.sub.B1, .sup.1J.sub.C,H=165 Hz), 97.9 (C-1.sub.A, .sup.1J.sub.C,H=170 Hz), 92.4, 92.3, 92.0 (4C, CCl.sub.3), 75.8 (C-3.sub.B), 75.0 (C-5.sub.A), 74.7 (C-3.sub.A, C-3.sub.B1), 72.9 (C-4.sub.A1), 72.4, 72.3 (CH.sub.2Bn), 72.1 (C-3.sub.A1), 71.4 (C-4.sub.A1), 69.5 (C-5.sub.B1), 68.8 (C-5.sub.B), 67.6 (CH.sub.2Bn-6), 67.5 (CH.sub.2Bn-6), 66.3 (OCH.sub.2), 65.4, 65.1 (2C, C-4.sub.B, C-4.sub.B1), 55.4 (2C, C-2.sub.B, C-2.sub.B1), 53.9, 51.3 (2C, C-2.sub.A, C-2.sub.A1), 48.1 (CH.sub.2N.sub.3), 29.0 (CH.sub.2,linker), 17.3, 17.1 (2C, C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.63H.sub.69Cl.sub.12N.sub.14O.sub.19 1745.1120, found 1745.1117.

    [0661] 3-Azidopropyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (60b). A solution of disaccharide donor 9b/10b (694 mg, 608 mol, 1.05 equiv.) and azidopropyl tetrasaccharide 59b (1.0 g, 579 mol, 1.0 equiv.) in anhyd. DCE (15 mL) was stirred with freshly activated 4 MS (1.2 g) for 1 h at rt under an Ar atmosphere. The reaction mixture was cooled to 15 C. and TMSOTf (8 L, 43 mol, 0.07 equiv.) was added. After 40 min at 15 C., Et.sub.3N (12 L, 0.1 equiv.) was added. After stirring for 10 min, the suspension was filtered by passing through a pad of Celite, solids were washed with DCM, and the filtrate was concentrated under vacuum. Flash chromatography (Tol/ACN 84:14.fwdarw.85:15) gave hexasaccharide 60b (850 mg, 317 mol, 55%) as a white solid. The azidopropyl glycoside 60b had R.sub.f 0.3 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.104H.sub.106Cl.sub.18N.sub.19O.sub.28 2700.1824, found 2700.1892.

    [0662] 3-Azidopropyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (61b). A solution of tetrasaccharide donors 13b/14b (1.07 g, 547 mol, 1.05 equiv.) and azidopropyl tetrasaccharide 59b (900 mg, 521 mol, 1.0 equiv) in anhyd. DCE (22 mL) was stirred with freshly activated 4 MS (1.5 g) for 1 h at rt under an Ar atmosphere. The reaction mixture was cooled to 5 C. and TMSOTf (7 L, 38 mol, 0.07 equiv.) was added. After stirring for 40 min at 5 C., Et.sub.3N (8 L, 0.1 equiv.) was added. After stirring for 10 min, solids were filtered and washed with DCM. The filtrate was concentrated under vacuum. Flash chromatography (Tol/ACN 84:14.fwdarw.84:16) of the residue gave azidopropyl octasaccharide 61b (850 mg, 317 mol, 55%) as a white solid. The coupling product 61b had R.sub.f 0.25 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.134H.sub.139Cl.sub.24N.sub.25O.sub.37 1770.6082, found 1770.6075.

    [0663] Additional Routes to Linker-Equipped Glycosides [0664] Cbz-masked aminopropyl linker [0665] Acetal-masked ketone-encompassing linker

    ##STR00109##

    [0666] 2-Methyl-1,3-dioxolane-2-ethyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (62b). A solution of donor 9b/10b (550 mg, 482 mol, 1.0 equiv) and 2-methyl-1,3-dioxolane-2-ethanol (127 mg, 963 mol, 2.0 equiv) in anhyd. DCE (8 mL) was stirred with freshly activated 4 MS (800 mg) for 30 min at rt under an Ar atmosphere. The reaction mixture was cooled to 15 C. and TMSOTf (6.0 L, 34 mol, 0.07 equiv.) was added. After stirring for 1 h at 15 C., the mixture was quenched with Et.sub.3N (6 L, 0.1 equiv.), filtered, and concentrated. Flash chromatography using Tol/EtOAc (80:20) gave the desired disaccharide 62b (410 mg, 405 mol, 78%) as a white solid. Disaccharide 62b had R.sub.f 0.4 (Tol/EtOAc 4:1) and HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.47H.sub.53Cl.sub.6N.sub.6O.sub.12, 1103.1821; found 1103.1847.

    [0667] 3-(Benzyloxycarbonylamino)-1-propyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (63b). A solution of donor 9b/10b (600 mg, 525 mol, 1.0 equiv.) and 3-(benzyloxycarbonylamino)-1-propanol (220 mg, 1.05 mmol, 2.0 equiv.) in anhyd. DCE (9 mL) was stirred with freshly activated 4 MS (800 mg) for 30 min at rt under an Ar atmosphere. The reaction mixture was cooled to 15 C. and TMSOTf (7.0 L, 61 mol, 0.07 equiv.) was added. After stirring for 40 min at 15 C., Et.sub.3N (7 L, 0.1 equiv.) was added to the reaction mixture, which was stirred at rt for 10 min. The suspension was filtered and concentrated. Flash chromatography (Tol/EtOAc 7:3) gave the desired disaccharide 63b (560 mg, 481 mol, 91%) as a white solid. Disaccharide 63b had R.sub.f 0.35 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.52H.sub.56Cl.sub.6N.sub.7O.sub.12, 1180.2113; found 1180.2091.

    Example 10. Linker-Equipped Oligosaccharides Featuring a B-Endchain Residue

    [0668] Azidopropyl Aglycon as Linker Precursor

    ##STR00110##

    [0669] Allyl 4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (9c). A solution of alcohol 8 (6.0 g, 16.1 mmol, 1.0 equiv) in anhyd. DMF (40 mL) was cooled to 0 C. under an Ar atmosphere. Benzyl bromide (2.3 mL, 19.3 mmol, 1.2 equiv.) was added followed by the portionwise addition of NaH (60% in mineral oil, 1.16 g, 48.3 mol, 3.0 equiv.). After stirring for 24 h at 0 C., diluted aq. NH.sub.4Cl (200 mL) was added and the aq. layer was washed with DCM (200 mL) twice. The combined organic parts were dried over Na.sub.2SO.sub.4, filtered and the filtrate was concentrated under reduced pressure. Flash chromatography of crude residue (Tol/EtOAc 90:10-84:16) gave the desired benzyl ether 9c (4.1 mg, 8.87 mmol, 55%) as a white solid. Further elution (Tol/EtOAc 90:10-60:40) gave the unreacted starting material 8 (1.6 g, 26%) corresponding to a 75% corrected yield. The benzyl ether 9c had R.sub.f 0.6 (Tol/EtOAc 7:3). .sup.1H NMR (CDCl.sub.3) 7.36-7.28 (m, 5H, H.sub.Ar), 7.08 (d, 1H, J.sub.NH,2=7.0 Hz, NH), 5.91-5.81 (m, 1H, CH.sub.All), 5.28-5.24 (m.sub.po, 1H, CH.sub.2All), 5.19-5.16 (m.sub.po, 1H, CH.sub.2All), 4.93 (d, 1H, J.sub.1,2=8.6 Hz, H-1), 4.70 (d, 1H, J=10.8 Hz, CH.sub.2Bn), 4.62 (d, 1H, CH.sub.2Bn), 4.50 (dd, 1H, J.sub.2,3=10.6 Hz, J.sub.3,4=3.6 Hz, H-3), 4.35-4.30 (m.sub.po, 1H, CH.sub.2All), 4.09-4.04 (m.sub.po, 1H, CH.sub.2All), 3.75 (brd, 1H, H-4), 3.69-3.59 (m.sub.po, 2H, H-2, H-5), 1.35 (d, 3H, J.sub.5,6=6.0 Hz, H-6). .sup.13C NMR (CDCl.sub.3) 162.0 (CO.sub.NHTCA), 137.0 (C.sub.q,Ar), 133.6 (CH.sub.An), 128.6, 128.3 (C.sub.Ar), 117.9 (CH.sub.2All), 97.7 (C-1, .sup.1J.sub.C,H=163 Hz), 92.4 (CCl.sub.3), 76.1 (C-3), 72.6 (CH.sub.2Bn), 70.2 (CH.sub.2All), 69.0 (C-5), 63.0 (C-4), 56.0 (C-2), 17.6 (C-6). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C18H.sub.25Cl.sub.3N.sub.5O.sub.4 480.0967; found 480.0967.

    [0670] 4-Azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy-/-D-galactopyranose (10c). [Ir(COD)(PMePh.sub.2).sub.2]PF.sub.6 (200 mg, 237 mol, 0.03 equiv.) was dissolved in anhyd. THF (10 mL) and stirred for 30 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed repeatedly with Ar and transferred by means of a cannula into a solution of allyl glycoside 9c (3.65 g, 7.89 mmol, 1.0 equiv.) in anhyd. THF (70 mL). After 2 h stirring at rt, a TLC analysis (Tol/EtOAc 85:15) showed the complete disappearance of the starting material and presence of a slightly less polar spot. NIS (2.1 g, 9.47 mmol, 1.2 equiv.) in THF/H.sub.2O (1:1, 20 mL) was added. After stirring for 1 h at rt, a TLC analysis (Tol/EtOAc 7:3) revealed the full consumption of the isomerized intermediate and the presence of a more polar spot. 10% Aq. Na.sub.2SO.sub.3 was added and volatiles were evaporated. The aq. phase was extracted with DCM (100 mL) three times. The combined organic layers were washed with water and brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under vacuum. Purification of the residue by flash chromatography (Tol/EtOAc 80:20.fwdarw.70:30) yielded the expected /-hemiacetal 10c (3.1 g, 7.34 mmol, 93%) as a white floppy solid. Hemiacetal 10c had R.sub.f 0.5, 0.2 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.15H.sub.21Cl.sub.3N.sub.5O.sub.4 440.0654; found 440.0652.

    [0671] 4-Azido-3-O-benzyl-2,4,6-trideoxy-2-trichloroacetamido-/-D-galactopyranosyl (N-phenyl)trifluoroacetimidate (11c). Hemiacetal 10c (3.0 g, 7.10 mmol, 1.0 equiv.) was dissolved in acetone (35 mL). PTFA-Cl (1.46 mL, 9.24 mmol, 1.3 equiv.) was added followed by Cs.sub.2CO.sub.3 (2.7 g, 8.53 mmol, 1.2 equiv.). After stirring for 2 h at rt, the reaction mixture was filtered, washed with acetone and the filtrate was concentrated under reduced pressure. The crude donor was purified by passing through a short silica column (cHex/EtOAc 90:10) to give the PTFA donor 11c as a mixture of /-isomers (3.9 g, 6.57 mmol, 92.5%). The -isomer had R.sub.f 0.7 (Tol/EtOAc 9:1). .sup.1H NMR (CDCl.sub.3) 7.44-7.35 (m, 5H, H.sub.Ar), 7.32-7.24 (m, 2H, H.sub.Ar), 7.14-7.11 (tt.sub.po, 1H, H.sub.Ar), 6.79 (d, 2H, J=7.6 Hz, H.sub.Ar), 6.46 (d, 1H, J.sub.2,NH=7.7 Hz, NH), 6.39 (brs, 1H, H-1), 4.82 (d, 1H, J=12.0 Hz, CH.sub.2Bn), 4.62 (d.sub.po, 1H, CH.sub.2Bn), 4.59-4.53 (m.sub.po, 1H, H-2), 4.07-4.05 (m.sub.po, 1H, H-5), 3.99-3.96 (m.sub.po, 2H, H-3, H-4), 1.36 (d, 1H, J.sub.5,6=6.0 Hz, H-6). .sup.13C NMR (CDCl.sub.3) 161.8 (CO.sub.NHTCA), 142.9, 135.5 (C.sub.q,Ar), 129.3, 128.9, 128.8, 128.5, 128.1, 126.3, 124.6, 120.5, 119.2 (C.sub.Ar), 119.2 (CF.sub.3), 94.1 (C-1.sub.A, .sup.1J.sub.C,H=191 Hz), 92.1 (CCl.sub.3), 75.0 (C-3), 71.6 (CH.sub.2Bn), 67.7 (C-5), 61.9 (C-4), 50.2 (C-2), 17.4 (C-6). HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.23H.sub.21Cl.sub.3F.sub.3N.sub.5O.sub.4Na, 616.0503; found 616.0496.

    [0672] Allyl 4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (12c). The PTFA donor 11c (3.4 g, 5.73 mmol, 1.0 equiv.) and acceptor 7 (4.99 g, 5.73 mmol, 1.0 equiv.) were mixed and dried over vacuum. The dried mass dissolved in anhyd. DCM (100 mL) was stirred with freshly activated 4 MS (6.0 g) for 30 min at rt under an Ar atmosphere. The reaction mixture was cooled to 30 C. and TMSOTf (51 L, 287 mol, 0.05 equiv.) was added. After 40 min at 30 C., a TLC analysis (Tol/EtOAc 4:1) showed the absence of PTFA donor and the presence of a new spot. Et.sub.3N was added and after 15 min, the suspension was filtered and volatiles were evaporated. Flash chromatography (Tol/ACN 90:10.fwdarw.88:12) yielded the desired trisaccharide 12c (3.8 g, 3.1 mmol, 52%) as a white solid. Along with the undesired -isomer (2.6 g, 3.64 mmol, 35%). The desired product 12c had R.sub.f 0.4 (Tol/ACN 4:1). .sup.1H NMR (CDCl.sub.3) 7.43-7.15 (m, 15H, H.sub.Ar), 7.05 (d, 1H, J.sub.2,NH=7.1 Hz, NH.sub.B1), 6.76 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.51 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.A), 5.89-5.80 (m, 1H, CH.sub.All), 5.31-5.18 (m.sub.po, 4H, CH.sub.2All, CH.sub.2Bn-6), 5.17 (d.sub.po, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A), 4.90 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B1), 4.78 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.73 (d, 1H, J.sub.4,5=2.7 Hz, H-5.sub.A), 4.68-4.62 (m.sub.po, 3H, CH.sub.2Bn, H-3.sub.B), 4.54 (dd.sub.po, 1H, J.sub.2,3=10.6 Hz, J.sub.4,5=3.4 Hz, H-3.sub.B1), 4.47 (dd.sub.po, 2H, J=12.6 Hz, CH.sub.2Bn), 4.33-4.29 (m.sub.po, 2H, H-4.sub.A, CH.sub.2All), 4.05-4.01 (m.sub.po, 1H, CH.sub.2All), 3.95-3.86 (m.sub.po, 3H, H-2.sub.A, H-3.sub.A, H-4.sub.B), 3.66 (d, 1H, J.sub.3,4=3.4 Hz, H-4.sub.B1), 3.61-3.46 (m.sub.po, 4H, H-2.sub.B, H-2.sub.B1, H-5.sub.B, H-5.sub.B1), 1.30 (d, 3H, J.sub.5,6=6.2 Hz, H-6.sub.B1), 1.27 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.2 (C-6.sub.A), 162.0, 161.9, 161.6 (CO.sub.NHTCA), 137.1, 137.0, 135.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 129.0 (2C), 128.8, 128.7, 128.6, 128.3 (3C), 125.3 (C.sub.Ar), 117.9 (CH.sub.2All), 99.0 (C-1.sub.B1, .sup.1J.sub.C,H=165 Hz), 98.2 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 97.8 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.5, 92.3 (3C, CCl.sub.3), 75.5 (C-3.sub.B1), 75.2 (C-5.sub.A), 74.8 (C-3.sub.B), 73.0 (C-3.sub.A), 72.7 (CH.sub.2Bn), 72.4 (CH.sub.2Bn), 71.8 (C-4.sub.A), 70.0 (CH.sub.2All), 69.0 (C-5.sub.B), 68.8 (C-5.sub.B1), 67.5 (CH.sub.2Bn-6), 65.4 (C-4.sub.B), 63.0 (C-4.sub.B1), 55.8, 55.6 (2C, C-2.sub.B, C-2.sub.B1), 54.1 (C-2.sub.A), 17.5, 17.3 (2C, C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.48H.sub.54Cl.sub.9N.sub.10O.sub.13 1293.1063; found 1293.1068.

    [0673] 4-Azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranose (13c). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (123 mg, 145 mol, 0.05 equiv.) was dissolved in anhyd. THF (8 mL) and stirred for 20 min under an H.sub.2 atmosphere. The resulting yellow solution was degassed with a continuous flow of Ar and poured into a solution of allyl glycoside 12c (3.7 g, 2.90 mmol, 1.0 equiv.) in anhyd. THF (50 mL). After stirring for 2 h at rt, NIS (783 mg, 3.48 mmol, 1.2 equiv.) in THF/H.sub.2O was added. After stirring for another 1 h at rt, 10% aq. sodium sulphite was added until full decoloration. Volatiles were evaporated and the aq. phase was extracted with DCM (50 mL) three times. The organic phases were combined, washed with brine, dried over anhyd. Na.sub.2SO.sub.4 and concentrated. Purification by flash chromatography (Tol/EtOAc 75:25.fwdarw.60:40) yielded the / hemiacetal 13c (3.3 g, 2.67 mmol, 92%) as a white solid. The /-anomer had R.sub.f 0.5, 0.25 (Tol/EtOAc 75:25.fwdarw.60:40). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.45H.sub.50Cl.sub.9N.sub.10O.sub.13 1257.0713; found 1253.0750.

    [0674] 4-Azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl (N-phenyl)trifluoroacetimidate (14c). PTFACl (550 L, 3.47 mmol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (1.04 g, 3.20 mmol, 1.2 equiv.) were added to hemiacetal 13c (3.3 g, 2.67 mmol, 1.0 equiv.) dissolved in acetone (27 mL). After stirring for 2 h at rt, the reaction mixture was filtered through a pad of Celite and washed with acetone (5 mL) twice. The filtrate was concentrated under reduced pressure and the crude PTFA donor was purified by passing through a short pack of silica (cHex/EtOAc 75:25.fwdarw.65:35). The /-PTFA donor 14c was isolated as a solid (3.25 g, 2.31 mmol, 86% over two steps). The PTFA donor had R.sub.f 0.75, 0.8 (Tol/EtOAc 7:3). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.53H.sub.54Cl.sub.9F.sub.3N.sub.11O.sub.13 1424.1046; found 1424.1029.

    [0675] Allyl 4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1-4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (15c). The PTFA trisaccharide 14c (1.0 g, 711 mol, 1.0 equiv.) and disaccharide acceptor 7 (619 mg, 711 mmol, 1.0 equiv.) were stirred with freshly activated 4 MS (1.0 g) in anhyd. DCE (17 mL) for 1 h under an Ar atmosphere at rt. After cooling to 10 C., TMSOTf (7.7 L, 43 mol, 0.06 equiv.) was added and stirring was continued for 45 min while keeping the temperature of the reaction bath at 10 C. At completion, Et.sub.3N (10 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (6 mL) thrice. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 90:10.fwdarw.86:14) to give pentasaccharide 15c as a white solid (1.2 g, 574 mol, 84%). The coupling product 15c had R.sub.f 0.55 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.78H.sub.83Cl.sub.15Cl.sub.4N.sub.15O.sub.22 2114.1070; found 2114.1078. .sup.1H NMR (CDCl.sub.3) 7.41-7.17 (m, 25H, H.sub.Ar), 7.11 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B2), 7.02 (d, 1H, J.sub.2,NH=6.7 Hz, NH.sub.B1), 6.76 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.49 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.A1), 6.44 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.A), 5.89-5.80 (m, 1H, CH.sub.All), 5.36 (d, 1H, J=12.0 Hz, CH.sub.2Bn-6), 5.28-5.22 (m.sub.po, 3H, CH.sub.2All, CH.sub.2Bn-6), 5.20-5.15 (m.sub.po, 3H, CH.sub.2All, CH.sub.2Bn-6, H-1.sub.A), 5.02 (d, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A1), 4.94 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B2), 4.88 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.B1), 4.81 (dd.sub.po, J.sub.2,3=10.6 Hz, J.sub.3,4=3.6 Hz, H-3.sub.B1), 4.78 (d.sub.po, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.72 (d.sub.po, 1H, J.sub.4,5=2.7 Hz, H-5.sub.A), 4.68 (d.sub.po, 1H, J.sub.4,5=2.6 Hz, H-5.sub.A1), 4.67-4.63 (m.sub.po, 3H, H-3.sub.B, CH.sub.2Bn), 4.58 (dd.sub.po, 1H, J.sub.2,3=10.6 Hz, J.sub.3,4=3.4 Hz, H-3.sub.B2), 4.51 (d, 1H, J=12.2 Hz, CH.sub.2Bn), 4.45-4.40 (m.sub.po, 3H, CH.sub.2Bn), 4.34-4.28 (m.sub.po, 2H, CH.sub.2All, H-4.sub.A1), 4.21 (t.sub.po, 1H, H-4.sub.A), 4.06-4.00 (m.sub.po, 1H, CH.sub.2All), 3.98-3.94 (m.sub.po, 1H, H-2.sub.A1), 3.91 (brd, 1H, J.sub.3,4=3.4 Hz, H-4.sub.B), 3.87-3.85 (m.sub.po, 3H, H-2.sub.A, H-3.sub.A, H-4.sub.B1), 3.81 (dd, 1H, J.sub.2,3=10.5 Hz, J.sub.3,4=2.5 Hz, H-3.sub.A1), 3.66 (brd, 1H, J.sub.3,4=3.0 Hz, H-4.sub.B2), 3.62-3.39 (m.sub.po, 6H, H-2.sub.B1, H-2.sub.B2, H-2.sub.B3, H-5.sub.B1, H-5.sub.B2, H-5.sub.B3), 1.30 (d, 1H, J.sub.5,6=6.2 Hz, H-6.sub.B), 1.27 (d, 1H, J.sub.5,6=6.4 Hz, H-6.sub.B), 1.22 (d, 1H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.2, 168.1 (C-6.sub.A), 162.1, 161.9, 161.5 (5C, CO.sub.NHTCA), 137.1, 137.0, 135.1, 135.0 (C.sub.q,Ar), 133.5 (CH.sub.All), 129.1 (2C), 129.0, 128.9, 128.2, (2C) (C.sub.Ar), 117.9 (CH.sub.2All), 99.0 (C-1.sub.B2, .sup.1J.sub.C,H=166 Hz), 98.6 (C-1.sub.B1, .sup.1J.sub.C,H=166 Hz), 98.4 (C-1.sub.A1, .sup.1J.sub.C,H=16 Hz), 98.0 (C-1.sub.A, .sup.1J.sub.C,H=168 Hz), 97.8 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 92.5, 92.4, 92.3 (5C, CCl.sub.3), 75.4 (C-5.sub.A, C-5.sub.A1, C-3.sub.B2), 74.6 (C-3.sub.B), 73.6 (C-3.sub.B1), 72.8 (2C, C-3.sub.A, C-3.sub.A1), 72.4 (2C, CH.sub.2Bn), 72.1 (CH.sub.2Bn), 71.9 (C-4.sub.A1), 71.7 (C-4.sub.A), 70.0 (CH.sub.2All), 69.3, 68.7 (C-5.sub.B, C-5.sub.B1, C-5.sub.B2), 67.5, 67.5 (2C, CH.sub.2Bn-6), 65.5 (C-4.sub.B), 65.3 (C-4.sub.B1), 63.1 (C-4.sub.B2), 55.9, 55.8, 55.6 (C-2.sub.B, C-2.sub.B1, C-2.sub.B2), 54.2, 53.8 (C-2.sub.A, C-2.sub.A1), 17.5, 17.3, 17.2 (C-6.sub.B, C-6.sub.B1, C-6.sub.B2).

    [0676] 4-Azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranose (16c). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (22 mg, 26 mol, 0.05 equiv.) was dissolved in anhyd. THF (6 mL) and stirred for 40 min under a H.sub.2 atmosphere. The resulting yellow solution was degassed with a continuous flow of Ar for 10 min and poured into a solution of allyl glycoside 15c (1.1 g, 527 mol, 1.0 equiv.) in anhyd. THF (5.0 mL). After stirring for 2 h at rt, NIS (142 mg, 632 mol, 1.2 equiv.) was added. After stirring for another 1 h at rt, the reaction was quenched with 10% aq. sodium sulphite. Volatiles were removed under vacuum and the aq. phase was extracted with DCM (320 mL). The organic phases were combined, washed with brine, dried over anhyd. Na.sub.2SO.sub.4 and concentrated. Purification by flash chromatography (Tol/EtOAc 70:30.fwdarw.60:40) yielded the expected /-hemiacetal 16c (1.0 g, 488 mol, 92%) as a white solid. The /-anomers had R.sub.f 0.55, 0.7 (Tol/EtOAc 1:1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.75H.sub.79C.sub.15N.sub.15O.sub.22 2066.0847; found 2066.0888.

    [0677] 4-Azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl (N-phenyl)trifluoroacetimidate (17c). Hemiacetal 16c (1.0 g, 488 mol, 1.0 equiv.) was dissolved in acetone (6.0 mL). PTFACl (100 L, 635 mol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (191 mg, 586 mol, 1.2 equiv.) were added. After stirring for 2 h at rt, the reaction mixture was filtered through a pad of Celite and solids were washed with acetone (6 mL) twice. The filtrate was concentrated under reduced pressure and the crude PTFA donor was purified by passing through a short pack of silica gel eluting with cHex/EtOAc (70:30.fwdarw.50:50). Fractions corresponding to the product were combined to give the expected PTFA donor 17c as a mixture of /-isomers (920 mg, 415 mol, 85%). HRMS (ESI.sup.+): calcd for C.sub.83H.sub.83Cl.sub.15F.sub.3N.sub.16O.sub.22 2239.1121; found 2239.11185.

    [0678] 3-Azidopropyl 4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (18c). The PTFA donor 14c (1.1 g, 782 mol, 1.0 equiv.) and 3-azido-1-propanol (144 L, 1.56 mmol, 2.0 equiv.) were stirred with freshly activated 4 MS (1.0 g) in anhyd. DCE (15.6 mL) for 1 h under an Ar atmosphere at rt. After cooling to 15 C., TMSOTf (10 L, 55 mol, 0.07 equiv.) was added and stirring was continued for 45 min while keeping the temperature of the reaction bath at 15 C. At completion, Et.sub.3N (12 L) was added. The suspension was filtered through a fitted funnel and washed with DCM (10 mL) twice. Flash chromatography (Tol/ACN 90:10.fwdarw.88:12) of the residue gave the azidopropyl trisaccharide 18c as a white solid (875 mg, 663 mol, 85%). The coupling product 18c had R.sub.f 0.45 (Tol/ACN 4:1). .sup.1H NMR (CDCl.sub.3) 7.43-7.17 (m, 15H, H.sub.Ar), 7.01 (d, 1H, J.sub.2,NH=6.8 Hz, NH), 6.74 (d, 1H, J.sub.2,NH=7.2 Hz, NH), 6.51 (d, 1H, J.sub.2,NH=6.8 Hz, NH), 5.30-5.19 (m.sub.po, 3H, CH.sub.2Bn-6, H-1.sub.A), 4.86 (d, 1H, J.sub.1,2=8.4 Hz, H-11), 4.73 (d.sub.po, 1H, J.sub.4,5=2.5 Hz, H-5.sub.A), 4.70 (d.sub.po, 1H, J.sub.1,2=8.1 Hz, H-1.sub.B), 4.64 (dd.sub.po, 2H, J=11.2 Hz, CH.sub.2Bn), 4.56 (dd, J.sub.2,3=10.8 Hz, J.sub.3,4=3.6 Hz, H-3.sub.B), 4.50-4.43 (m.sub.po, 3H, H-3.sub.B1, CH.sub.2Bn), 4.29 (t, 1H, J=2.2 Hz, H-4.sub.A), 3.94-3.85 (m.sub.po, 4H, OCH.sub.2, H-2.sub.A, H-3.sub.A, H-4.sub.B), 3.66 (d, 1H, J.sub.3,4=2.8 Hz, H-4.sub.B1), 3.64-3.50 (m.sub.po, 4H, OCH.sub.2, H-2.sub.B, H-2.sub.B1, H-5.sub.B), 3.47 (dq.sub.po, 1H, H-5.sub.B1), 3.35 (t, 2H, J=6.6 Hz, NCH.sub.2), 1.88-1.76 (m, 2H, CH.sub.2), 1.29 (d, 6H, J.sub.5,6=6.4 Hz, H-6.sub.B, H-6.sub.B1). .sup.13C NMR (CDCl.sub.3) 168.2 (C-6.sub.A), 161.9 (3C, CO.sub.NHTCA), 137.1, 137.0, 135.0 (C.sub.q,Ar), 129.0, 128.8 (2C), 128.7, 128.6, 128.3 (2C), 128.2 (C.sub.Ar), 99.1 (2C, C-1.sub.B1, C-1.sub.B, .sup.1J.sub.C,H=168 Hz, .sup.1J.sub.C,H=162 Hz), 98.0 (C-1.sub.A, .sup.1J.sub.C,H=168 Hz), 75.6 (C-3.sub.B1), 75.3 (C-5.sub.A), 74.9 (C-3.sub.B), 73.0 (C-3.sub.A), 72.7 (CH.sub.2Bn), 72.5 (CH.sub.2Bn), 71.9 (C-4.sub.A), 69.0, 68.9 (2C, C-5.sub.B, C-5.sub.B1), 67.6 (OCH.sub.2), 66.3 (CH.sub.2Bn-6), 65.3 (C-4.sub.B), 62.9 (C-4.sub.B1), 55.7, 55.3 (2C, C-2.sub.B, C-2.sub.B1), 54.1 (C-2.sub.A), 48.1 (NCH.sub.2), 29.0 (CH.sub.2), 17.5, 17.3 (2C, C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.48H.sub.55Cl.sub.9N.sub.13O.sub.13 1336.1234; found 1336.1229.

    [0679] 3-Azidopropyl 4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (19c). The PTFA donor 17c (920 mg, 415 mol, 1.0 equiv.) and 3-azido-1-propanol (76 L, 829 mol, 2.0 equiv.) were stirred with freshly activated 4 MS (600 mg) in anhyd. DCE (10 mL) for 1 h under an Ar atmosphere at rt. After cooling to 15 C., TMSOTf (5.0 L, 29 mol, 0.07 equiv.) was added and stirring was continued for 45 min while keeping the temperature of the reaction bath at 15 C. At completion, Et.sub.3N (10 L) was added. The suspension was filtered through a fitted funnel and washed with DCM. Volatiles were evaporated and the residue was purified by flash chromatography (Tol/ACN 90:10.fwdarw.88:12) to give the azidopropyl pentasaccharide 19c as a white solid (785 mg, 359 mol, 88%). The coupling product 19c had R.sub.f 0.4 (Tol/ACN 4:1).

    [0680] .sup.1H NMR (CDCl.sub.3) 7.34-7.15 (m, 25H, H.sub.Ar), 7.07 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.B2), 7.01 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B1), 6.76 (d, 1H, J.sub.2,NH=7.2 Hz, NH.sub.B), 6.50 (d, 1H, J.sub.2,NH=6.8 Hz, NH.sub.A1), 6.44 (d, 1H, J.sub.2,NH=8.0 Hz, NH.sub.A), 5.35 (d, 1H, J=12.0 Hz, CH.sub.2Bn-6), 5.28-5.17 (m.sub.po, 4H, H-1.sub.A, CH.sub.2Bn-6), 5.05 (d, 1H, J.sub.1,2=7.6 Hz, H-1.sub.A1), 4.91 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B2), 4.84 (d, 1H, J.sub.1,2=8.2 Hz, H-1.sub.B1), 4.78 (dd.sub.po, J.sub.2,3=10.6 Hz, J.sub.3,4=4.0 Hz, H-3.sub.B1), 4.72 (d.sub.po, 1H, J.sub.4,5=2.6 Hz, H-5.sub.A1), 4.70 (d.sub.po, J.sub.1,2=8.0 Hz, H-1.sub.B), 4.69 (d.sub.po, 1H, J.sub.4,5=2.8 Hz, H-5.sub.A), 4.65 (dd.sub.po, 2H, CH.sub.2Bn), 4.58-4.52 (m.sub.po, 2H, H-3.sub.B2, H-3.sub.B), 4.49-4.40 (m.sub.po, 4H, CH.sub.2Bn), 4.31 (t.sub.po, 1H, J.sub.4,5=J.sub.3,4=2.6 Hz, H-4.sub.A1), 4.21 (t.sub.po, 1H, H-4.sub.A), 4.00-3.86 (m.sub.po, 6H, H-2.sub.A, H-2.sub.A1, H-3.sub.A, H-3.sub.A, H-4.sub.B, H-4.sub.B1, OCH.sub.2), 3.66 (brd, 1H, J.sub.3,4=2.8 Hz, H-4.sub.B2), 3.65-3.51 (m.sub.po, 7H, OCH.sub.2,H-2.sub.B1, H-2.sub.B2, H-2.sub.B3, H-5.sub.B1, H-5.sub.B2, H-5.sub.B3), 3.36 (t, 2H, J=6.6 Hz, NCH.sub.2), 1.88-1.76 (m.sub.po, 2H, CH.sub.2), 1.30 (d, 1H, J.sub.5,6=6.2 Hz, H-6.sub.B), 1.28 (d, 1H, J.sub.5,6=6.4 Hz, H-6.sub.B), 1.22 (d, 1H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (CDCl.sub.3) 168.2, 168.1 (C-6.sub.A), 162.1, 161.9, 161.6 (5C, CO.sub.NHTCA), 137.1, 137.0, 135.1, 134.9 (C.sub.q,Ar), 129.1 (2C), 129.0, 128.9, 128.8, 128.7, 128.6 (2C), 128.5, 128.3 (2C) (C.sub.Ar), 99.2 (C-1.sub.B, .sup.1J.sub.C,H=161 Hz, C-1.sub.B2, .sup.1J.sub.C,H=163 Hz), 98.8 (C-1.sub.B1, .sup.1J.sub.C,H=167 Hz), 98.3 (C-1.sub.A1, .sup.1J.sub.C,H=168 Hz), 97.8 (C-1.sub.A, .sup.1J.sub.C,H=169 Hz), 92.5, 92.4, 92.3 (5C, CCl.sub.3), 75.4 (C-5.sub.A, C-5.sub.A1), 74.6 (C-3.sub.B1), 74.6 (C-3.sub.B, C-3.sub.B2), 72.9 (C-3.sub.A, C-3.sub.A1), 72.7 (CH.sub.2Bn), 72.5 (CH.sub.2Bn), 72.1 (CH.sub.2Bn), 71.9 (C-4.sub.A1), 71.7 (C-4.sub.A), 69.0, 68.8, 68.8 (C-5.sub.B, C-5.sub.B1, C-5.sub.B2), 67.5 (2C, CH.sub.2Bn-6), 66.2 (OCH.sub.2), 65.4 (C-4.sub.B), 65.3 (C-4.sub.B1), 63.0 (C-4.sub.B2), 55.8, 55.7, 55.6 (C-2.sub.B, C-2.sub.B1, C-2.sub.B2), 54.1, 53.8 (C-2.sub.A, C-2.sub.A1), 48.1 (NCH.sub.2), 29.0 (CH.sub.2), 17.5, 17.3, 17.2 (C-6.sub.B, C-6.sub.B1, C-6.sub.B2). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.78H.sub.84.sup.35Cl.sub.11.sup.37Cl.sub.4N.sub.18O.sub.22 2157.1242 found 2157.1284.

    Example 11. Full Deprotection of the 4.SUB.A.-Terminal Oligosaccharides to Provide the Corresponding Aminopropyl-Linker Equipped Oligosaccharides

    [0681] ##STR00111##

    [0682] 3-Aminopropyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (64b). Disaccharide 63b was subjected to hydrogenation-mediated full deprotection (protocol 1). Thus, 20% Pd(OH).sub.2C (180 mg, 3 equiv.) was added to the fully protected disaccharide 63b (100 mg, 86 mol) in 2-MeTHF/isopropanol/water (1:15:3, v/v/v, 39 mL). The suspension was stirred vigorously for 1 h under a hydrogen atmosphere, 0.6 mM aq. NaHCO.sub.3 (440 L, 3 equiv.) was added. After stirring for another hour under a hydrogen atmosphere, more NaHCO.sub.3 (150 L, 1 equiv.) was added (twice) and the reaction was run for three more hours at which time LC-MS indicated reaction completion. Another equivalent of NaHCO.sub.3 (150 L, 1 equiv.) was added to reach a total of 6 equivalent. The suspension was filtered by passing through a 0.2 m filter. Volatiles were evaporated and the residue was purified by C-18 chromatography eluting with water/ACN 100:0 to 90:10. The desired aminopropyl disaccharide 64b was obtained as a white lyophilized powder (23 mg, 48 mol, 56%). The free disaccharide 64b had RP-HPLC (215 nm/ESLD): R.sub.t=4.1/4.3 min (conditions A). .sup.1H NMR (D.sub.2O) 4.85 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A), 4.41 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.34-4.31 (m, 2H, H-4.sub.A, H-3.sub.B), 3.98-3.85 (m, 5H, H-5.sub.A, H-2.sub.A, H-5.sub.B H-2.sub.B, OCH.sub.2Pr), 3.71-3.66 (m.sub.po, 2H, H-3.sub.A, OCH.sub.2Pr), 3.42 (brs, 1H, H-4.sub.B), 3.06 (t, 2H, J=6.8 Hz, NCH.sub.2), 2.04, 1.99 (2s, 6H, CH.sub.3Ac), 1.93 (t, J=6.0 Hz, CH.sub.2), 1.28 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (D.sub.2O) 175.2, 174.3 (NHCO.sub.A,B), 174.2 (C-6.sub.A), 101.8 (C-1.sub.B, .sup.1J.sub.C,H=161.2 Hz), 100.5 (C-1.sub.A, .sup.1J.sub.C,H=165.2 Hz), 78.3 (C-5.sub.A), 77.2 (C-3.sub.B), 69.7 (C-5.sub.B), 69.0 (C-4.sub.A), 68.3 (C-3.sub.A), 68.0 (OCH.sub.2Pr), 53.6 (C-4.sub.B), 51.7 (C-2.sub.A), 50.7 (C-2.sub.B), 37.6 (NCH.sub.2), 26.8 (CH.sub.2Pr), 22.2 (2C, CH.sub.3Ac), 15.8 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.19H.sub.35N.sub.4O.sub.10, 479.2348; found 479.2346.

    [0683] 3-Aminopropyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (65b). The azidopropyl tetrasaccharide 58b was submitted to full hydrogenation-mediated deprotection (protocol 1). The free tetrasaccharide 65b had RP-HPLC (215 nm/ELSD): R.sub.t=5.0/5.2 min (conditions A). .sup.1H NMR (D.sub.2O) 4.90 (d, 1H, J.sub.1,2=8.0 Hz, H-1.sub.A), 4.81-4.74 (m.sub.po, 2H, H-1.sub.A1, H-1.sub.B), 4.52-4.46 (m.sub.po, 3H, H-5.sub.A, H-5.sub.A1, H-1.sub.B1), 4.40-4.36 (m.sub.po, 2H, H-4.sub.A, H-4.sub.A1), 4.18-4.16 (m, 2H, H-3.sub.B, H-3.sub.B1), 4.10-4.05 (m, 2H, H-5.sub.B, H-5.sub.B1), 4.01-3.99 (m.sub.po, 3H, H-4.sub.B, H-2.sub.A, OCH.sub.2Pr), 3.90-3.80 (m, 4H, H-2.sub.A, H-4.sub.B, H-2.sub.B, H-2.sub.B1), 3.76-3.65 (m.sub.po, 4H, H-2.sub.B, H-3.sub.A, H-3.sub.A1, OCH.sub.2Pr), 3.11-3.07 (m.sub.po, 2H, NCH.sub.2), 2.06, 2.03, 2.02, 1.98 (2s, 6H, CH.sub.3Ac), 1.98-1.91 (m.sub.po, 2H, CH.sub.2), 1.33 (d.sub.po, 6H, J.sub.5,6=6.4 Hz, H-6.sub.B, H-6.sub.B1). .sup.13C NMR (D.sub.2O) 174.9, 174.7, 174.5, 174.4 (4C, NHCO.sub.A,B), 174.1, 174.2 (2C, C-6.sub.A, C-6.sub.A1), 103.0 (C-1.sub.B1, .sup.1J.sub.C,H=166.0 Hz), 101.7 (C-1.sub.B, .sup.1J.sub.C,H=162.5 Hz), 101.1 (C-1.sub.A, .sup.1J.sub.C,H=165.0 Hz), 100.9 (C-1.sub.A, .sup.1J.sub.C,H=165.5 Hz), 78.0, 77.6 (2C, C-5.sub.A, C-5.sub.A1), 77.5 (C-4.sub.A), 76.0, 75.7 (2C, C-3.sub.B, C-3.sub.B1), 69.3 (C-4.sub.A), 67.5, 67.3 (2C, C-5.sub.B, C-5.sub.B1), 68.2 (OCH.sub.2Pr), 68.2, 67.9 (2C, C-3.sub.A, C-3.sub.A1), 54.9, 54.8 (2C, C-4.sub.B, C-4.sub.B1), 51.6, 51.5 (2C, C-2.sub.A, C-2.sub.A1), 51.0, 50.9 (2C, C-2.sub.B, C-2.sub.B1), 37.6 (NCH.sub.2), 26.7 (CH.sub.2Pr), 22.5, 22.4 (4C, CH.sub.3Ac), 15.7, 15.6 (2C, C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.35H.sub.60N.sub.7O.sub.19 882.3938, found 882.3929.

    [0684] 3-Aminopropyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (66b). Protocol 3: Hexasaccharide 60b (111 mg, 4.1 mol) was solubilized in 2-MeTHF (2.0 mL) and diluted to 0.26 mM per repeating unit in 2-MeTHF/iPrOH/H.sub.2O (1:10:1 v/v/v, 50 mL). The solution was degassed and 10% Pd/C (220 mg) was added. The suspension was stirred vigorously under a hydrogen atmosphere for 4 h, at which time 10% Pd/C (220 mg) and 1 M aq. NaHCO.sub.3 (25 L, 30 mol, 7 equiv.) were added. After 40 h, 10% Pd/C (220 mg) was added and the suspension was stirred under a hydrogen atmosphere for 3 days, at which time RP-HPLC and HRMS controls revealed the presence of the desired compound and absence of any major chlorinated intermediate. The suspension was passed through a pad of Celite, and solids were washed extensively with water. Volatiles were evaporated and the aqueous phase was lyophilized, the residue solubilized in water (2.0 mL). pH was adjusted to 6 by addition of 1 M aq. NaHCO.sub.3 and the was passed through a 0.2 m filter and the filtrate was lyophilized. RP-HPLC purification (ACN in 0.08% TFA 0-18%) of the crude material gave the desired aminopropyl hexasaccharide 66b (22 mg, 17 mol, 41%) as a white lyophilized powder. The free hexasaccharide 66b had RP-HPLC (215 nm/ELSD): R.sub.t=6.6/6.7 min (conditions A), R.sub.t=6.0/6.1 min (conditions E). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.51H.sub.85N.sub.10O.sub.28 1285.5529; found 1285.5510, m/z [M+2H].sup.2+ calcd for C.sub.51H.sub.86N.sub.10O.sub.28 643.2801; found 643.2792.

    [0685] 3-Aminopropyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (67b). Protocol 3: Octasaccharide 61b (77 mg, 2.2 mol) was solubilized in 2-MeTHF (1.5 mL) and diluted to 0.19 mM per repeating unit in 2-MeTHF/iPrOH/H.sub.2O (1:10:1 v/v/v, 27 mL). The solution was degassed and 10% Pd/C (164 mg, 7 equiv.) was added. The suspension was stirred vigorously under a hydrogen atmosphere for 4 h, at which time 10% Pd/C (164 mg, 7 equiv.) and 1 M aq. NaHCO.sub.3 (18 L, 18 mol, 8 equiv.) were added. After 40 h, 10% Pd/C (164 mg, 7 equiv.) was added and the suspension was stirred under a hydrogen atmosphere for 3 days, at which time RP-HPLC and HRMS controls revealed the presence of the desired compound and of minor amounts of monochlorinated intermediates. The suspension was passed through a pad of Celite, and solids were washed extensively with water. Volatiles were evaporated and the aqueous phase was lyophilized. The residue solubilized in water (2.0 mL) was passed through a 0.2 m filter and the filtrate was lyophilized. RP-HPLC purification (ACN in 0.08% TFA 0-18%) of the crude material (53 mg) in water (2.5 mL, pH 5) gave the desired aminopropyl octasaccharide 67b (6.5 mg, 3.8 mol, 17%) as a white lyophilized powder. The free octasaccharide 67b had RP-HPLC (215 nm/ELSD): R.sub.t=7.6/7.8 min (conditions A), R.sub.t=6.2/6.4 min (conditions D). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.67H.sub.111N.sub.13O.sub.37 844.8596; found 844.8588, m/z [M+3H].sup.3+ calcd for C.sub.67H.sub.112N.sub.13O.sub.37 563.5755; found 563.5750.

    Example 12. Aminopropyl Linker Modification into Conjugation-Ready Oligosaccharides

    [0686] Linker-Modification with SAMA or SPDP: Chemoselective Introduction of a Masked Thiol

    ##STR00112##

    [0687] 3-(2-Methylthioacetyl)ethylamido)-propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (68b). The aminopropyl oligosaccharide 64b (3.1 mg, 6.5 mol, 1.0 equiv.) was dissolved in 0.1 M phosphate buffer pH 6.2 (800 L). S-Acetylthioglycolic acid pentafluorophenyl ester (1.94 mg, 6.5 mol, 1.0 equiv.) in ACN (60 L) was added portionwise over 2 h to the reaction mixture stirred at rt. Progress was monitored by RP-HPLC and LCMS analysis. As some starting material was still visible after 2.5 h, the reaction was left at rt overnight. Purification by RP-HPLC (0 to 20% gradient over 20 min, UV detection at 230 nm) and repeated lyophilization of the pooled fractions of interest provided disaccharide 68b (1.5 mg, 39%) as a lyophilized powder. The desired linker equipped product disaccharide 68b had RP-HPLC (215 nm/ESLD): R.sub.t=8.84/8.98 min (conditions D). .sup.1H NMR (D.sub.2O) 4.87 (d, 1H, J.sub.1,2=8.7 Hz, H-1.sub.A), 4.50 (brs, 1H, H-5.sub.A), 4.44 (d, 1H, J.sub.1,2=8.6 Hz, H-1.sub.B), 4.36 (brs, 1H, H-4.sub.A), 4.12 (dd, 1H, J.sub.2,3=10.4 Hz, J.sub.3,4=4.0 Hz, H-3.sub.B), 4.04 (q.sub.po, 1H, J.sub.5,6=6.4 Hz, H-5.sub.B), 3.98-3.92 (m.sub.po, 1H, H-2.sub.A), 3.93 (brd, 1H, H-4.sub.B), 3.89-3.83 (m.sub.po, 1H, OCH.sub.2Pr), 3.81-3.76 (t.sub.po, 1H, H-2.sub.B), 3.66 (dd.sub.po, 1H, H-3.sub.A), 3.61 (brs, 2H, CH.sub.2,SAc), 3.61-3.56 (m.sub.po, 2H, OCH.sub.2Pr), 3.27-3.12 (m, 2H, NCH.sub.2), 2.39 (SCH.sub.3), 2.01, 1.98 (2s, 6H, CH.sub.3Ac), 1.72 (t.sub.po, J=6.4 Hz, CH.sub.2Pr), 1.28 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.23H.sub.39N.sub.4O.sub.12S, 595.2280; found 595.2270. HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.23H.sub.38N.sub.4O.sub.12SNa 617.2099; found 617.2087.

    [0688] General procedure for the introduction of the-(2-pyridyldithio)propionamide moiety: The aminopropyl oligosaccharide (1-10 mg, 1.0 equiv.) was dissolved in 0.1 M phosphate buffer pH 6.2 (1.0 mg/100-300 L). 3-(2-Pyridyldithio)propionic acid N-hydroxysuccinimide ester (1.0 equiv.) in DMSO (1 mg in 10 L) was added. The reaction mixture was stirred for 6-16 h at rt. Progress was monitored by RP-HPLC and LCMS analysis. At completion, the desired product was purified by RP-HPLC using a gradient of ACN in 0.08% aq. TFA as eluent. The product was confirmed based on NMR and HRMS analysis.

    [0689] 3-(3-(2-Pyridyldithio)propionamido)-propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (69b). Disaccharide 64b (15 mg, 31 mol, 1.0 equiv.) dissolved in 0.1 M phosphate buffer pH 6.2 (3.1 mL) was subjected to regioselective amidation. SPDP (990 g, 31 mol, 1.0 equiv.) was added and the reaction mixture was stirred for 6 h. RP-HPLC of the crude material gave the amide 69b (5.1 mg, 24%) was obtained as a white lyophilized solid. The linker-equipped disaccharide 69b had RP-HPLC (215 nm/ELSD): R.sub.t=11.9/12.1 min (conditions D). .sup.1H NMR (D.sub.2O) 8.55 (d, 1H, J=5.2 Hz, H.sub.Ar), 8.20 (t, 1H, J=4.0 Hz, H.sub.Ar), 8.07 (d, 1H, J=8.4 Hz, H.sub.Ar), 7.62 (t, 1H, J=6.4 Hz, H.sub.Ar), 4.87 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A), 4.54 (brs, 1H, H-5.sub.A), 4.45 (d, 1H, J.sub.1,2=8.6 Hz, H-1.sub.B), 4.36 (brs, 1H, H-4.sub.A), 4.13 (dd, 1H, J.sub.2,3=10.8 Hz, J.sub.3,4=4.0 Hz, H-3.sub.B), 4.02 (q.sub.po, 1H, J.sub.5,6=6.4 Hz, H-5.sub.B), 3.96 (t.sub.po, 1H, J=8.6 Hz, H-2.sub.A), 3.91 (brd, 1H, H-4.sub.B), 3.87-3.83 (m.sub.po, 1H, OCH.sub.2), 3.77 (t, 1H, H-2.sub.B), 3.67 (dd.sub.po, 1H, H-3.sub.A), 3.60-3.54 (m, 1H, OCH.sub.2), 3.21-3.09 (m, 2H, NCH.sub.2), 3.11 (t, 2H, J=6.8 Hz, COCH.sub.2), 2.65 (t, 2H, SCH.sub.2), 2.00, 1.97 (2s, 6H, CH.sub.3Ac), 1.71 (t, 2H, J=6.8 Hz, CH.sub.2), 1.28 (d, 3H, J.sub.5,6=6.4 Hz, H-6.sub.B). .sup.13C NMR (D.sub.2O) 174.6, 174.0 (NHCO.sub.A,B), 173.4 (NHCO), 173.1 (C-6.sub.A), 156.7 (C.sub.Ar,q), 144.7, 143.4, 124.4, 12.4 (C.sub.Ar), 101.5 (C-1.sub.B, .sup.1J.sub.C,H=161.4 Hz), 101.0 (C-1.sub.A, .sup.1J.sub.C,H=165.2 Hz), 76.7 (C-5.sub.A), 76.2 (C-3.sub.B), 68.6 (C-4.sub.A), 68.0 (OCH.sub.2Pr), 67.9 (C-3.sub.A), 67.3 (C-5.sub.B), 54.7 (C-4.sub.B), 51.4 (C-2.sub.A), 50.8 (C-2.sub.B), 36.2 (NCH.sub.2), 34.5 (SCH.sub.2), 34.2 (COCH.sub.2), 28.3 (CH.sub.2Pr), 22.2 (2C, CH.sub.3Ac), 15.5 (C-6.sub.B). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.27H.sub.42N.sub.5O.sub.11S, 2676.2317; found 676.2292.

    [0690] 3-(3-(2-Pyridyldithio)propionamido)-propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (70b). The crude tetrasaccharide 65b (20 mol theo. from 58b) was dissolved in 0.1 M phosphate buffer pH 6.2 (4.5 mL) and stirred vigorously at rt. A solution of SPDP (4.7 mg, 15 mol, 0.75 equiv.) in DMSO (50 L) was added portionwise over 2 h. After overnight stirring, more SPDP (4.7 mg, 15 mol, 0.75 equiv.) in DMSO (50 L) was added and the reaction mixture was stirred for another 6 h. One fourth of the total volume was purified by RP-HPLC. The amide 70b (5.1 mg, 24% from 58b) was obtained as a white lyophilized solid. The linker-equipped tetrasaccharide 70b had RP-HPLC (215 nm/ELSD): R.sub.t=10.7/10.8 min (conditions E). .sup.1H NMR (D.sub.2O) 8.56 (d, 1H, J=5.2 Hz, H.sub.Ar), 8.23 (t, 1H, J=8.0 Hz, H.sub.Ar), 8.09 (d, 1H, J=8.8 Hz, H.sub.Ar), 7.65 (t, 1H, J=6.4 Hz, H.sub.Ar), 4.87 (d, 1H, J.sub.1,2=8.2 Hz, H-1.sub.A), 4.78-4.75 (m.sub.po, 2H, H-1.sub.A1, H-1.sub.B), 4.63 (brs, 1H), 4.57 (brs, 1H, H-5.sub.A1), 4.44-4.42 (m.sub.po, 2H, H-5.sub.A, H-1.sub.B1), 4.37 (d.sub.po, 1H, H-4.sub.A), 4.16-4.10 (m, 2H, H-3.sub.B), 4.04-4.00 (m, 2H, H-5.sub.B, H-5.sub.B1), 3.97-3.95 (m.sub.po, 1H, H-2.sub.A), 3.91 (brs, 1H, H-4.sub.B), 3.88-3.66 (m.sub.po, 7H, H-2.sub.A, H-2.sub.B, H-2.sub.B1, H-3.sub.A, H-3.sub.A1, H-4.sub.B, OCH.sub.2Pr), 3.58-3.56 (m.sub.po, 1H, OCH.sub.2Pr), 3.19-3.10 (m.sub.po, 4H, NCH.sub.2, COCH.sub.2-linker), 2.64 (t, J=6.4 Hz, SCH.sub.2), 2.06, 1.99, 1.95 (3s, 12H, CH.sub.3Ac), 1.71-1.68 (m.sub.po, 2H, CH.sub.2), 1.29 (d.sub.po, 6H, J.sub.5,6=6.2 Hz, H-6.sub.B). .sup.13C NMR (D.sub.2O) 174.6, 174.4, 173.9 (4C, NHCO.sub.A,B), 173.3 (NHCO.sub.linker), 172.9, 172.7 (C-6.sub.A), 162.7 (C.sub.Ar,q), 156.5, 144.3, 143.8, 124.5, 123.5 (C.sub.Ar), 102.9 (C-1.sub.B1, .sup.1J.sub.C,H=166.4 Hz), 101.6 (C-1.sub.B, .sup.1J.sub.C,H=162.8 Hz), 101.0 (2C, C-1.sub.A, C-1.sub.A1, .sup.1J.sub.C,H=166.0 Hz, 167.0 Hz), 76.8, 76.6 (3C, C-4.sub.A, C-5.sub.A, C-5.sub.A1), 76.0, 75.9 (2C, C-3.sub.B, C-1.sub.B1), 68.5 (C-4.sub.A), 68.0 (OCH.sub.2Pr), 67.8, 67.6 (2C, C-5.sub.B) C-1.sub.B1), 67.4, 67.3 (2C, C-3.sub.A, C-3.sub.A1), 54.8, 54.7 (2C, C-4.sub.B, C-4.sub.B1), 51.5, 51.3 (2C, C-2.sub.A, C-1.sub.A1), 50.9, 50.8 (2C, C-2.sub.B, C-1.sub.B1), 36.2 (NCH.sub.2), 34.4 (SCH.sub.2), 34.2 (COCH.sub.2), 28.3 (CH.sub.2Pr), 22.3, 22.2 (4C, CH.sub.3Ac), 15.6, 15.5 (2C, C-6.sub.B, C-1.sub.B1). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.43H.sub.67N.sub.8O.sub.20S.sub.2 1079.3908, found 1079.3873.

    [0691] 3-(3-(2-Pyridyldithio)propionamido)-propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (71b). A solution of hexasaccharide 66b (5.1 mg, 3.97 mol, 1.0 equiv.) in 0.1 M phosphate buffer pH 6.2 (1.0 mL) was stirred vigorously at rt and a solution of succinimidyl 3-(2-pyridyldithio)propionate (SPDP, 1.25 mg, 3.97 mol, 1.0 equiv.) in DMSO (30 L) was added to it portionwise (10 L every 30 min). After stirring for 2 h at rt, more SPDP (0.4 mg, 0.3 equiv.) in DMSO (10 L) was added and the reaction mixture was stirred for another 1.5 h at rt, then overnight at 4 C. RP-HPLC purification (0.fwdarw.40% ACN in 0.08% aq. TFA over 20 min) of the reaction mixture followed by lyophilisation gave the linker-equipped hexasaccharide 71b (2.7 mg, 1.82 mol, 46%) and some remaining aminopropyl glycoside 66b (1.45 mg, 1.13 mol) to reach a corrected yield of 64%. The desired 71b had RP-HPLC (215 nm/ELSD): R.sub.t=11.3/11.4 min (conditions D). .sup.1H NMR (D.sub.2O, 800 MHz) 8.56 (dd, 1H, J=5.8, <1.0 Hz, H.sub.Ar), 8.23 (dt, 1H, J=8.0, 1.6 Hz, H.sub.Ar), 8.07 (d, 1H, J=8.2 Hz, H.sub.Ar), 7.64 (m.sub.po, 1H, J=6.4 Hz, H.sub.Ar), 4.83 (d, 1H, J.sub.1,2=8.3 Hz, H-1.sub.A2), 4.73-4.66 (peaks overlap with HOD, 4H, H-1.sub.A1, H-1.sub.A, H-1.sub.B2, H-1.sub.B1), 4.62-4.60 (m.sub.po, 2H, H-5.sub.A1, H-5.sub.A), 4.54 (d, 1H, J.sub.4,5=2.2 Hz, H-5.sub.A2), 4.41 (t.sub.po, 1H, J.sub.3,4=J.sub.4,5=2.5 Hz, H-4.sub.A1), 4.40 (t.sub.po, 1H, J.sub.3,4=J.sub.4,5=2.6 Hz, H-4.sub.A), 4.38 (d.sub.po, 1H, J.sub.1,2=8.5 Hz, H-1.sub.B), 4.33 (t.sub.po, 1H, J.sub.3,4=J.sub.4,5=2.7 Hz, H-4.sub.A2), 4.11-4.06 (m.sub.po, 3H, H-3.sub.B2, H-3.sub.B1, H-3.sub.B), 4.00-3.96 (m, 3H, H-5.sub.B2, H-5.sub.B1, H-5.sub.B), 3.93 (dd, 1H, J.sub.2,3=10.6 Hz, J.sub.1,2=8.4 Hz, H-2.sub.A2), 3.87 (d, 1H, J.sub.3,4=4.4 Hz, H-4.sub.B), 3.83-3.75 (m.sub.po, 7H, H-2.sub.A1, H-2.sub.A, H-2.sub.B2, H-2.sub.B1, H-4.sub.B2, H-41, OCH.sub.2Pr), 3.72 (dd, 1H, J.sub.2,3=10.8 Hz, J.sub.1,2=8.8 Hz, H-2.sub.B), 3.70-3.68 (m.sub.po, 2H, H-3.sub.A1, H-3.sub.A), 3.62 (dd, 1H, J.sub.2,3=10.8 Hz, J.sub.3,4=3.1 Hz, H-3.sub.A2), 3.53-3.50 (m.sub.po, 1H, OCH.sub.2Pr), 3.14-3.11 (m, 1H, NCH.sub.2), 3.08-3.05 (m.sub.po, 1H, NCH.sub.2), 3.07 (t.sub.po, 2H, J=6.6 Hz, COCH.sub.2-linker), 2.60 (t, 2H, J=6.6 Hz, SCH.sub.2-linker), 1.96, 1.95, 1.94, 1.91, 1.90 (6s, 18H, CH.sub.3Ac), 1.68-1.63 (m, 2H, CH.sub.2Pr), 1.26-1.24 (m.sub.po, 9H, H-6.sub.B2, H-6.sub.B1, H-6.sub.B). .sup.13C NMR (D.sub.2O, 800 MHz) 174.6, 174.4, 174.4, 173.9, 173.3 (NHCO.sub.A,B), 173.3 (NHCO.sub.linker), 172.7, 172.5, 172.4 (C-6.sub.A), 162.8 (C.sub.Ar,q), 156.5, 144.1, 143.8, 124.6, 123.6 (C.sub.Ar), 102.9 (C-1.sub.B2, .sup.1J.sub.C,H=165 Hz), 102.9 (C-1.sub.B1, .sup.1J.sub.C,H=166 Hz), 101.5 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 101.1 (C-1.sub.A2, .sup.1J.sub.C,H=166 Hz), 101.0 (C-1.sub.A, .sup.1J.sub.C,H=166 Hz), 101.0 (C-1.sub.A1, .sup.1J.sub.C,H=168 Hz), 76.7, 76.6 (2C, C-4.sub.A, C-4.sub.A1), 76.5, 76.4 (3C, C-5.sub.A2, C-5.sub.A1, C-5.sub.A), 76.0, 75.8 (3C, C-3.sub.B2, C-3.sub.B1, C-3.sub.B), 68.4 (C-4.sub.A2), 67.9 (OCH.sub.2Pr), 67.8, 67.5 (3C, C-3.sub.A2, C-3.sub.A1, C-3.sub.A), 67.3, 67.2 (3C, C-5.sub.B2, C-5.sub.B1, C-5.sub.B), 54.8 (3C, C-4.sub.B2, C-4.sub.B1, C-4.sub.B), 51.4, 51.2 (3C, C-2.sub.A2, C-2.sub.A1, C-2.sub.A), 50.9, 50.8 (3C, C-2.sub.B2, C-2.sub.B1, C-2.sub.B), 36.1 (NCH.sub.2), 34.4 (SCH.sub.2), 34.2 (COCH.sub.2), 28.2 (CH.sub.2Pr), 22.2, 22.1 (6C, CH.sub.3Ac), 15.5 (3C, C-6.sub.B2, C-6.sub.B1, C-6.sub.B). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.59H.sub.93N.sub.11O.sub.29S.sub.2 741.7786; found 741.7782. HRMS (ESI.sup.+): m/z [M+3H].sup.3+ calcd for C.sub.59H.sub.94N.sub.11O.sub.29S.sub.2 494.8548; found 494.8546.

    [0692] 3-(3-(2-Pyridyldithio)propionamido)-propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (72b). A solution of octasaccharide 67b (5.1 mg, 3.0 mol, 1.0 equiv.) in 0.1 M phosphate buffer pH 6.4 (1.0 mL) was stirred vigorously at rt and a solution of succinimidyl 3-(2-pyridyldithio)propionate (SPDP, 950 g, 3.0 mol, 1.0 equiv.) in DMSO (30 L) was added to it portionwise (10 L every 30 min). After stirring for 3 h at rt, more SPDP (90 g, 0.3 equiv.) in DMSO (5.0 L) was added and the reaction mixture was stirred for another 30 min at rt. RP-HPLC purification (0.fwdarw.40% ACN in 0.08% aq. TFA over 20 min) followed by lyophilisation gave the linker-equipped octasaccharide 72b (2.3 mg, 1.82 mol, 41%) and some remaining aminopropyl glycoside 67b (2.0 mg, 1.16 mol) to reach a corrected yield of 67%. The desired 72b had RP-HPLC (215 nm/ELSD): R.sub.t=11.0/11.2 min (conditions D). .sup.1H NMR (D.sub.2O, 800 MHz) 8.56 (d, 1H, J=5.8 Hz, H.sub.Ar), 8.15 (tt.sub.po, 1H, J=8.0 Hz, H.sub.Ar), 8.01 (d, 1H, J=8.4 Hz, H.sub.Ar), 7.56 (tt.sub.po, 1H, J=6.0 Hz, H.sub.Ar), 4.83 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.A3), 4.74-4.66 (peaks overlap, 6H, H-1.sub.A2, H-1.sub.A1, H-1.sub.A, H-1.sub.B3, H-1.sub.B2, H-1.sub.B1), 4.56-4.55 (m.sub.po, 3H, H-5.sub.A2, H-5.sub.A1, H-5.sub.A), 4.54 (brs, 1H, H-5.sub.A3), 4.40-4.38 (m.sub.po, 4H, H-4.sub.A2, H-4.sub.A1, H-4.sub.A, H-1.sub.B), 4.33 (t, 1H, J.sub.3,4=J.sub.4,5=2.5 Hz, H-4.sub.A3), 4.11-4.06 (m.sub.po, 4H, H-3.sub.B3, H-3.sub.B2, H-3.sub.B1, H-3.sub.B), 4.00-3.96 (m.sub.po, 4H, H-5.sub.B3, H-5.sub.B2, H-5.sub.B1, H-5.sub.B), 3.93 (dd, 1H, J.sub.2,3=10.7 Hz, J.sub.1,2=8.5 Hz, H-2.sub.A3), 3.87 (d, 1H, J.sub.3,4=4.4 Hz, H-4.sub.B), 3.83-3.74 (m.sub.po, 10H, H-2.sub.A2, H-2.sub.A1, H-2.sub.A, H-2.sub.B3, H-2.sub.B2, H-2.sub.B1, H-4.sub.B3, H-4.sub.B2, H-4.sub.B1, OCH.sub.2Pr), 3.72 (dd, 1H, J.sub.2,3=11.0 Hz, J.sub.1,2=8.9 Hz, H-2.sub.B), 3.69-3.67 (m.sub.po, 3H, H-3.sub.A2, H-3.sub.A1, H-3.sub.A), 3.62 (dd, 1H, J.sub.2,3=10.6 Hz, J.sub.3,4=2.9 Hz, H-3.sub.A3), 3.54-3.51 (m.sub.po, 1H, OCH.sub.2Pr), 3.14-3.11 (m, 1H, NCH.sub.2), 3.08-3.04 (m.sub.po, 1H, NCH.sub.2), 3.06 (t.sub.po, 2H, J=6.3 Hz, COCH.sub.2-linker), 2.60 (t, 2H, J=6.4 Hz, SCH.sub.2-linker), 1.96, 1.95, 1.94, 1.92, 1.91, 1.90 (8s, 24H, CH.sub.3Ac), 1.67-1.63 (m, 2H, CH.sub.2), 1.26-1.24 (m.sub.po, 12H, H-6.sub.B3, H-6.sub.B2, H-6.sub.B1, H-6.sub.B). .sup.13C NMR (D.sub.2O, 800 MHz) 174.6, 174.4, 174.4, 174.1 (8C, NHCO.sub.A,B), 173.3 (NHCO.sub.linker), 173.2, 172.9, 172.4 (4C, C-6.sub.A), 162.8 (C.sub.Ar,q), 156.8, 144.8, 143.1, 124.0, 123.3 (C.sub.Ar), 103.0 (C-1.sub.B3, .sup.1J.sub.C,H=165 Hz), 102.9 (C-1.sub.B2, C-1.sub.B1, .sup.1J.sub.C,H=165 Hz, 166 Hz), 101.6 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 101.1 (C-1.sub.A3, .sup.1J.sub.C,H=165 Hz), 101.0 (3C, C-1.sub.A2, C-1.sub.A1, C-1.sub.A, .sup.1J.sub.C,H=166 Hz, 168 Hz), 76.9 (3C, C-4.sub.A2, C-4.sub.A1, C-4.sub.A), 76.8, 76.7 (4C, C-5.sub.A3, C-5.sub.A2, C-5.sub.A1, C-5.sub.A), 76.0, 75.7, 75.6 (4C, C-3.sub.B3, C-3.sub.B2, C-3.sub.B1, C-3.sub.B), 68.6 (C-4.sub.A3), 68.2 (OCH.sub.2Pr), 67.9, 67.8, 67.6 (4C, C-3.sub.A3, C-3.sub.A2, C-3.sub.A1, C-3.sub.A), 67.3, 67.2 (4C, C-5.sub.B3, C-5.sub.B2, C-5.sub.B1, C-5.sub.B), 54.8, 54.3 (4C, C-43, C-4.sub.B2, C-4.sub.B1, C-4.sub.B), 51.4, 51.3 (4C, C-2.sub.A3, C-2.sub.A2, C-2.sub.A1, C-2.sub.A), 50.9, 50.8 (4C, C-2.sub.B3, C-2.sub.B1, C-2.sub.B1, C-2.sub.B), 36.1 (NCH.sub.2), 34.4 (SCH.sub.2), 34.0 (COCH.sub.2), 28.2 (CH.sub.2Pr), 22.2, 22.1 (8C, CH.sub.3Ac), 15.5 (4C, C-6.sub.B3, C-6.sub.B2, C-6.sub.B1, C-6.sub.B). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.75H.sub.118N.sub.14O.sub.38S.sub.2 943.3581; found 943.3567. HRMS (ESI.sup.+): m/z [M+3H].sup.3+ calcd for C.sub.75H.sub.119N.sub.14O.sub.38S.sub.2 629.2412; found 629.2402.

    Example 13. Glycerol Aglycon Modification into Conjugation-Ready Oligosaccharides

    [0693] Linker-Modification with PDPH: Chemoselective Introduction of a Masked Thiol by Means of an Aldehyde Intermediate

    ##STR00113##

    [0694] 2-Oxoethyl (2-acetamido-2-deoxy-4-O-methyl--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (53d). Diol 53 (10.8 mg, 21.2 mol, 1.0 equiv.) was dissolved in water (2.6 mL) and aq. NaIO.sub.4 (87 mM, 246 L, 4.58 mg, 21.2 mol, 1.0 equiv.) was added. The reaction mixture was stirred in the dark at rt and reaction progress was monitored by RP-HPLC. After 2 h, more aq. NaIO.sub.4 (50 L, 930 g, 4.3 mol, 0.2 equiv.) was added as some remaining starting 53 was still visible (RP-HPLC (215 nm): R.sub.t=7.1 min (conditions A)) and stirring went on for 30 min at rt. The crude material was purified by RP-HPLC (0.fwdarw.70% ACN in water) to give the desired aldehyde 53d (7.9 mg, 16.5 mol, 77%). Disaccharide 53d had RP-HPLC (215 nm): R.sub.t=6.8 min (conditions A), R.sub.t=5.9 min (conditions E). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.19H.sub.39N.sub.3O.sub.11, 478.2031; found 478.2012.

    [0695] 2-(3-(2-Pyridyldithio)propanoylhydrazono)ethyl (2-acetamido-2-deoxy-4-O-methyl--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (53e). Aldehyde 53d (5.1 mg, 10.7 mol, 1.0 equiv.) was dissolved in water/0.1 M Phosphate buffer pH 5.5 (5:1, 1.2 mL) and 3-(2-pyridyldithio)propionyl hydrazide (PDPH, 4.9 mg, 21.4 mol, 2 equiv.) in DMSO (20 L) was stirred vigorously for 3 h at rt and reaction progress was monitored by RP-HPLC and LCMS. The crude material, showing MS (ESI.sup.+) for C.sub.27H.sub.40N.sub.6O.sub.11S.sub.2Na m/z [M+H]+ 711.2, was purified by RP-HPLC (0.fwdarw.70% ACN in water) to give the desired hydrazone 53e (5.1 mg, 7.4 mol, 69%). Linker-equipped disaccharide 53e had RP-HPLC (215 nm): R.sub.t=10.0 min (conditions E). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.27H.sub.41N.sub.6O.sub.11S.sub.2 689.2275, found 689.2270.

    [0696] 2-(Biotine-hydrazide)-ethyl (2-acetamido-2-deoxy-4-O-methyl--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (53f). Diol 53 (2.0 mg, 3.9 mol, 1.0 equiv.) was dissolved in acetate buffer pH 5 (1.3 mL) and a solution of aq. NaIO.sub.4 (20 mM, 195 L, 830 g, 3.9 mol, 1.0 equiv.) was added. The reaction mixture was stirred in the dark for 2 h at rt. Aq. KCl (1 M, 1.5 mg, 20 L, 20 mol, 5.1 equiv.) was added and the reaction mixture was stirred for an additional 20 min. (+)-Biotin hydrazide (2.0 mg, 8.2 mol, 2.0 equiv.) was added and the mixture was stirred at rt for 48 h. The crude mixture was lyophilized and the residue was purified by RP-HPLC. The desired biotinylated disaccharide 53f (0.3 mg, 0.4 mol, 10%) had HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.29H.sub.48N.sub.7O.sub.12S, 718.3076; found 718.3073.

    [0697] 2-Oxoethyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (54d). Diol 54 (9.2 mg, 10.1 mol, 1.0 equiv.) was dissolved in water (1.5 mL) and aq. NaIO.sub.4 (87 mM, 139 L, 2.58 mg, 12.1 mol, 1.2 equiv.) was added. The reaction mixture was stirred in the dark at rt and reaction progress was monitored by RP-HPLC (conditions A). After 2 h, the starting 54 (R.sub.t=8.9 min) was absent and a new product was visible (R.sub.t=8.4 min, MS (ESI.sup.+) for calcd for C.sub.35H.sub.57N.sub.6O.sub.20: m/z [M+H]+ 881.3). The crude material was purified by RP-HPLC (0.fwdarw.20% ACN in water) to give the desired aldehyde 54d (7.9 mg, 9.0 mol, 89%). Tetrasaccharide 54d had RP-HPLC (215 nm): R.sub.t=9.8 min (conditions A).

    [0698] 2-(3-(2-Pyridyldithio)propanoylhydrazono)ethyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (54e). Aldehyde 54d (4.3 mg, 4.9 mol, 1.0 equiv.) was dissolved in water/0.1 M Phosphate buffer pH 7.5 (4:1, 1.0 mL) and PDPH (2.2 mg, 21.4 mol, 2 equiv.) in DMSO (10 L) was stirred vigorously for 3 h at rt and reaction progress was monitored by RP-HPLC and LCMS showing the starting material eluting at R.sub.t=8.0 min (conditions A) and the product eluting at R.sub.t=17.1 min (conditions A), R.sub.t=9.9 min (conditions E), MS (ESI.sup.+) for C.sub.43H.sub.66N.sub.9O.sub.20S.sub.2: m/z [M+H]+ 1092.2, m/z [M+2H].sup.2+546.7. The crude material was purified by RP-HPLC (0.fwdarw.70% ACN in water) to give the desired hydrazone 54e (3.1 mg, 2.8 mol, 5%). Linker-equipped tetrasaccharide 54e had RP-HPLC (215 nm): R.sub.t=20.3 min (conditions A), R.sub.t=12.2/13.2 min (conditions E, sample in water/ammonium acetate buffer pH 6.6). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.43H.sub.66N.sub.9O.sub.20S.sub.21092.3860; found 1092.3860, m/z [M+2H].sup.2+ calcd for C.sub.43H.sub.67N.sub.9O.sub.20S.sub.2 546.6966; found 546.6968.

    Example 14. Oligosaccharide-Protein Conjugates Exemplified for Tetanus Toxoid as the Carrier and the Thiol-Maleimide Conjugation Chemistry by Use of the SPDP Strategy Starting from (AB).SUB.n .Oligosaccharides Equipped with a Linker at their Reducing End

    [0699] ##STR00114##

    [0700] General Method for the Conjugation Step

    [0701] Size Exclusion Chromatography (SEC). An KTA pure chromatography system (GE Healthcare Life Sciences) was equipped with a high-resolution preparative gel Hiload 16/600 Superdex 200 g column eluting with 0.2 m sterile filtered phosphate buffered saline (PBS) xl, pH 7.2, 1.0 mL/min for preparative chromatography or with a Superdex 200 Increase 3.2/300 eluting with PBS xl, 0.05 mL/min for analytical chromatography.

    [0702] Buffer exchange and concentration. Buffer exchange and concentration was performed using spin filters (Millipore, Amicon ultra 4 and 15) with a 30 kDa MWCO (15 min, 5,000 xg, rt). For each buffer exchange at least four consecutive cycles were performed, with an exchange ratio of at least 15 per cycle.

    [0703] Protein concentration is estimated by UV detection (=280 nm) for tetanus toxoid conjugates ((TT)==189,460 M.sup.1.Math.cm.sup.1, mw(TT): 150,551 D) and CRM 197 conjugates (((CRM)==54,570 M.sup.1.Math.cm.sup.1, mw(CRM): 58,413 D) with buffer as control

    [0704] Protocol for MALDI analysis. The protein solution (20 L) was passed through a ZipTip C4 and eluted on a MTP 384 ground steel target plate (Bruker-Daltonics, Germany) with 2 L of 20 mg/mL sinapinic acid in 50% aq. ACN containing 0.1% aq. TFA as the matrix solution. Samples were air-dried for 15 min. Data were acquired on a Bruker UltrafleXtrem instrument, using the Flexcontrol software (Bruker-Daltonics, Germany). 10,000 shots were recorded in the positive ion linear mode in the m/z range of 30-210 kDa.

    [0705] Disaccharide-TT conjugates (78b) from thiol-equipped disaccharide 73b. SEC-purified tetanus toxoid (TT, 150 kD, from Bio Farma (Bandung, Indonesia), 15.3 mg/mL, 228 L, 3.49 mg) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30 kD centrifugal filter four times. A solution of GMBS (1.04 mg, 3.71 mol, 160 equiv.) in DMSO (15 L) was added to the obtained solution of TT (23 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.1 containing 5 mM EDTA, and finally concentrated to reach a final concentration of intermediate 77b of 16.8 mg/mL.

    [0706] Disaccharide 69b (1.65 mg, 2.4 mol) was dissolved in 0.1 M phosphate buffer pH 6.1 (100 L). A 11.4 M solution of tris(2-carboxyethyl)phosphine hydrochloride (TCEP.Math.HCl) in 0.1 M phosphate buffer pH 6.1 (10 L, 672 g, 2.34 mol, 0.96 equiv.) was added and the solution was stirred at rt. Monitoring was achieved by RP-HPLC (conditions D) and HRMS revealing the absence of the starting 69b (R.sub.t=10.0 min) and the presence of a major product corresponding to the expected thiol 73b. The later had RP-HPLC (215 nm/ELSD): R.sub.t=8.3 min (conditions E). Thiol 73b was found to dimerize upon storage to have HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.44H.sub.75N.sub.8O.sub.22S.sub.2 1131.4437; found 1131.4430.

    [0707] Two individual portions of the stock solutions of intermediates 77b and 73b, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. Volumes of harvest were adjusted to a volume of 1.0 mL. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0708] Conjugate 78b1: From the stock solution of modified TT (77b, 89 L, 1.5 mg, 10 nmol) and crude thiol 73b (27 L, 404 g, 185 nmol, 60 equiv.).

    [0709] Conjugate 78b2: From the stock solution of modified TT (77b, 89 L, 1.5 mg, 10 nmol) and crude thiol 73b (18 L, 269 g, 530 nmol, 40 equiv.).

    [0710] Conjugate 78b3: From the stock solution of modified TT (77b, 89 L, 1.5 mg, 10 nmol) and crude thiol 73b (18 L, 269 g, 530 nmol, 30 equiv.).

    [0711] Tetrasaccharide-TT conjugates (79b) from thiol-equipped tetrasaccharide 74b. SEC-purified tetanus toxoid (TT, 150 kD, 1.9 mL, 6.17 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30kD centrifugal filter four times and finally concentrated to 860 L (final concentration of TT: 13.2 mg/mL). A solution of GMBS (3.4 mg, 12.1 mol, 160 equiv.) in DMSO (30 L) was added to the obtained solution of TT (88 M, 76 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.1 containing 5 mM EDTA, and finally concentrated to 810 L to reach a final concentration of intermediate 77b of 13.78 mg/mL.

    [0712] Tetrasaccharide 70b (3.68 mg, 3.41 mol) was dissolved in 0.1 M phosphate buffer pH 6.1 (300 L). A 64 mM solution of tris(2-carboxyethyl)phosphine hydrochloride (TCEP.Math.HCl) in DMSO (53 L, 980 g, 3.41 mol, 1.0 equiv.) was added and the solution was stirred at rt. Monitoring was achieved by RP-HPLC (conditions D/E) and HRMS revealing the absence of the starting 70b (R.sub.t=11.2/10.7 min) and the presence of a major product corresponding to the expected thiol 74b. The later had RP-HPLC (215 nm): R.sub.t=8.6 min (conditions D), R.sub.t=8.3 min (conditions E). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.38H.sub.63N.sub.7O.sub.20S, 970.3921; found 970.3917. HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.38H.sub.62N.sub.7O.sub.20SNa 992.3741; found 992.3735.

    [0713] Two individual portions of the stock solutions of intermediates 77b and 74b, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. Volumes of harvest were adjusted to a volume of 1.0 mL. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0714] Conjugate 79b1: From the stock solution of modified TT (77b, 145 L, 2.0 mg, 13.3 nmol) and crude thiol 74b (29.5 L, 287 nmol, 21.6 equiv.).

    [0715] Conjugate 79b2: From the stock solution of modified TT (77b, 72.6 L, 1.0 mg, 6.6 nmol) and crude thiol 74b (27.1 L, 264 nmol, 40 equiv.).

    [0716] Conjugate 79b3: From the stock solution of modified TT (77b, 72.6 L, 1.0 mg, 6.6 nmol) and crude thiol 74b (40.1 L, 396 nmol, 60 equiv.).

    [0717] As part of another experiment to reach a final concentration of intermediate 77b of 14.94 mg/mL, conjugates 79b4 and 79b5 were obtained.

    [0718] Conjugate 79b4: From the stock solution of modified TT (77b, 89 L, 1.3 mg, 8.7 nmol) and crude thiol 74b (31 L, 261 nmol, 30 equiv.).

    [0719] Conjugate 79b5: From the stock solution of modified TT (77b, 89 L, 1.3 mg, 8.7 nmol) and crude thiol 74b (remaining stock).

    [0720] Hexasaccharide-TT conjugates (80b) from thiol-equipped hexasaccharide 75b. SEC-purified tetanus toxoid (TT, 150 kD, 1.9 mL, 6.17 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30 kD centrifugal filter four times and finally concentrated to 860 L (final concentration of TT: 13.2 mg/mL). A solution of GMBS (3.4 mg, 12.1 mol, 160 equiv.) in DMSO (30 L) was added to the obtained solution of TT (88 M, 76 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.1 containing 5 mM EDTA, and finally concentrated to 810 L to reach a final concentration of intermediate 77b of 13.78 mg/mL.

    [0721] Hexasaccharide 71b (2.8 mg, 1.89 mol) was dissolved in 0.1 M phosphate buffer pH 6.1 (189 L). A 64 mM solution of tris(2-carboxyethyl)phosphine hydrochloride (TCEP.Math.HCl) in DMSO (30 L, 540 g, 1.89 mol, 1.0 equiv.) was added and the solution was stirred at rt. After 1 h, Monitoring by RP-HPLC (conditions D) and HRMS revealing the absence of the starting 71b (R.sub.t=11.3 min) and the presence of a major product corresponding to the expected thiol 75b. The later had RP-HPLC (215 nm): R.sub.t=8.6 min (conditions D). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.54H.sub.90N.sub.10O.sub.29S, 687.2792; found 687.2792. HRMS (ESI.sup.+): m/z [M+H+Na].sup.2+ calcd for C.sub.54H.sub.89N.sub.10O.sub.29SNa 698.2702; found 698.2699.

    [0722] Three individual portions of the stock solutions of intermediates 77b and 75b, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. Volumes of harvest were adjusted to a volume of 1.0 mL. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0723] Conjugate 80b1: From the stock solution of modified TT (77b, 72.6 L, 1.0 mg, 6.6 nmol) and crude thiol 75b (19.2 L, 166 nmol, 25 equiv.).

    [0724] Conjugate 80b2: From the stock solution of modified TT (77b, 65.3 L, 0.9 mg, 6.0 nmol) and crude thiol 75b (27.7 L, 239 nmol, 40 equiv.).

    [0725] Conjugate 80b3: From the stock solution of modified TT (77b, 65.3 L, 1.0 mg, 6.0 nmol) and crude thiol 75b (46.2 L, 398 nmol, 60 equiv.).

    [0726] Octasaccharide-TT conjugates (81b) from thiol-equipped octasaccharide 76b. SEC-purified tetanus toxoid (TT, 150 kD, 1.9 mL, 6.17 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30 kD centrifugal filter four times and finally concentrated to 860 L (final concentration of TT: 13.2 mg/mL). A solution of GMBS (3.4 mg, 12.1 mol, 160 equiv.) in DMSO (30 L) was added to the obtained solution of TT (88 M, 76 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.1 containing 5 mM EDTA, and finally concentrated to 810 L to reach a final concentration of intermediate 77b of 13.78 mg/mL.

    [0727] Octasaccharide 72b (1.0 mg, 530 mol) was dissolved in 0.1 M phosphate buffer pH 6.1 (100 L). A 64 mM solution of tris(2-carboxyethyl)phosphine hydrochloride (TCEP.Math.HCl) in DMSO (8.5 L, 152 g, 530 nmol, 1.0 equiv.) was added to reach a concentration of 4.9 nM of oligosaccharide and the solution was stirred at rt. After 1 h, Monitoring by RP-HPLC (conditions D) and HRMS revealing the absence of the starting 72b (R.sub.t=11.1 min) and the presence of a major product corresponding to the expected thiol 76b. The later had RP-HPLC (215 nm): R.sub.t=8.5 min (conditions D). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.70H.sub.115N.sub.13O.sub.38 888.8588; found 888.8589. HRMS (ESI.sup.+): m/z [M+H+Na].sup.2+ calcd for C.sub.70H.sub.114N.sub.13O.sub.38SNa, 899.8498; found 899.8496. HRMS (ESI.sup.+): m/z [M+2Na].sup.2+ calcd for C.sub.70H.sub.113N.sub.13O.sub.38SNa.sub.2 910.8407; found 910.8409.

    [0728] Different individual portions of various stock solutions of intermediates 77b and 76b, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. Volumes of harvest were adjusted to a volume of 1.0 mL. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0729] Conjugate 81b1: From the stock solution of modified TT (77b, 65.3 L, 0.9 mg, 6.0 nmol) and crude thiol 76b (24.4 L, 119 nmol, 20 equiv.).

    [0730] Conjugate 81b2: From the stock solution of modified TT (77b, 65.3 L, 0.9 mg, 6.0 nmol) and crude thiol 76b (48.9 L, 239 nmol, 40 equiv.).

    [0731] As part of another experiment to reach a final concentration of intermediate 77b of 13.85 mg/mL, conjugates 81b3-83b6 were obtained.

    [0732] Conjugate 81b3: From the stock solution of modified TT (77b, 75 L, 1.0 mg, 6.6 nmol) and crude thiol 76b (24 L, 104 nmol, 15 equiv.).

    [0733] Conjugate 81b4: From the stock solution of modified TT (77b, 319 L, 4.4 mg, 29.3 nmol) and crude thiol 76b (160 L, 705 nmol, 24 equiv.).

    [0734] Conjugate 81b5: From the stock solution of modified TT (77b, 255 L, 3.5 mg, 23.5 nmol) and crude thiol 76b (170 L, 750 nmol, 32 equiv.).

    [0735] Conjugate 81b6: From the stock solution of modified TT (77b, 28 L, 392 g, 2.6 nmol) and crude thiol 76b (23.6 L, 104 nmol, 32 equiv.).

    [0736] As part of another experiment to reach a final concentration of intermediate 77b of 12.8 mg/mL, conjugate 81b7 was obtained.

    [0737] Conjugate 81b7: From the stock solution of modified TT (77b, 936 L, 12.0 mg, 797 nmol) and crude thiol 76b obtained from precursor 72b (6.0 mg, 3.18 mol, 40 equiv.) featuring a masked thiol moiety.

    Example 15. Oligosaccharide-Protein Conjugates Exemplified for Tetanus Toxoid as the Carrier and the Thiol-Maleimide Conjugation Chemistry by Use of the PDPH Strategy

    [0738] ##STR00115##

    [0739] Disaccharide-TT conjugates (53h) from thiol-equipped disaccharide 53e. SEC-purified tetanus toxoid (TT, 150 kD, 6.12 mg/mL, 900 L, 5.5 mg) in PBS 1 was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30 kD centrifugal filter four times to reach a concentration of 10.9 mg/mL (550 L). A solution of GMBS (1.5 mg, 5.35 mol, 145 equiv.) in DMSO (15 L) was added to the obtained solution of TT (32.7 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged four times with 0.1 M Phosphate buffer pH 6.3 containing 5 mM EDTA, and finally concentrated to reach a final concentration of intermediate 77b of 14.2 mg/mL (380 L).

    [0740] Disaccharide 53e (4.6 mg, 6.67 mol) was dissolved in 0.1 M phosphate buffer pH 6.1 (165 p L). A 20 mM solution of TCEP.Math.HCl in 0.1 M phosphate buffer pH 6.3 (335 L, 1.91 mg, 6.66 mol, 1.0 equiv.) was added and the solution was stirred at rt for 1.5 h. Monitoring was achieved by RP-HPLC (conditions E) and LCMS revealing the absence of the starting 53e (R.sub.t=10.0 min) and the presence of a major product (R.sub.t=6.0 min) corresponding to the expected thiol 53g as revealed by MS (ESI.sup.+) for C.sub.22H.sub.37N.sub.5O.sub.11S: m/z [M+H]+ 580.2, m/z [M+Na].sup.+ 602.2. The stock solution corresponding to a total of 6.7 mol in 500 L was kept at 0 C. before use.

    [0741] Four individual portions of the stock solutions of intermediates 77b and 53g, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3.5 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. Volumes of harvest were adjusted to a volume of 1.0 mL. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0742] Conjugate 53h1: From the stock solution of modified TT (77b, 70 L, 1.0 mg, nmol) and crude thiol 53g (15 L, 115 g, 199 nmol, 30 equiv.).

    [0743] Conjugate 53h2: From the stock solution of modified TT (77b, 70 L, 1.0 mg, nmol) and crude thiol 53g (30 L, 230 g, 530 nmol, 60 equiv.)

    [0744] Conjugate 53h3: From the stock solution of modified TT (77b, 70 L, 1.0 mg, nmol) and crude thiol 53g (twice 15 L, 230 g, 530 nmol, 60 equiv.).

    [0745] Conjugate 53h4: From the stock solution of modified TT (77b, 89 L, 1.0 mg, nmol) and crude thiol 53g (thrice 15 L, 345 g, 597 nmol, 90 equiv.).

    TABLE-US-00001 TABLE 1 S. sonnei (AB).sub.n oligosaccharide-TT conjugates obtained by means of the thiol-maleimide chemistry. Average Code in Conjugate OS:TT ratio TT conc.sup.a OS conc Total FIGS. 1-7 number (m) mg/mL g/mL volume Ssonnei 53h1 4.7 1.68 SS-TT2 53h2 7.5 1.65 53h3 4 0.89 53h4 7.3 0.64 Son D 78b1 30 1.81 145 800 L Son C 78b2 23 1.66 100 1.1 mL Son B 78b3 13 1.59 56 870 L Ssonnei Son E 79b4 8.5 1.31 59 1.06 mL SS-TT4 Son F 79b5 12 1.58 95 1.01 mL Son G 79b1 11 2.03 100 920 L Son H 79b3 19 1.01 101 720 L Son N 79b2 13 2.27 158 875 L Ssonnei Son I 80b1 8.5 1.11 75 880 L SS-TT6 Son J 80b2 15 0.99 120 905 L Son K 80b3 19 0.94 145 580 L Ssonnei Son L 81b1 5.5 0.96 56 880 L SS-TT8 Son M 81b2 13 0.88 120 730 L Son W 81b3 7 1.05 80 910 L Son X 81b4 12.5 4.05 560 1150 L Son Y 81b5 13.5 2.75 400 1170 L Son Z 81b6 19 0.71 150 540 L Son AA 81b7 10 3.15 340 3.86 mL Ssonnei Son BA 82b1 6 2.15 175 800 L SS-TT10

    Example 16: Study in Mice

    [0746] Mice Immunization

    [0747] For each of the adjuvanted conjugates, seven week-old Balb/c female mice (Janvier Labs, France) were immunized intramuscularly (i.m.) with amounts of conjugates corresponding to 2.5, 2.0, 1.0 or 0.5 g equivalent of oligosaccharide per dose depending on the experiments, adjuvanted with aluminium hydroxide (alum, AlH, Alhydrogel, Brenntag, Denmark) unless stated. Alum was used at a concentration of 1.4 mg/mL in Tris pH 7.2 20 mM, and mixed v/v with the conjugates, resulting to a dose of 143 g per mouse/per injection. After 5 min incubation at rt, 200 L of the adjuvanted glycoconjugates were injected at two sites (100 L at each site). Three immunizations were performed at 3 week-interval. Blood samples were recovered one week after the third injection unless stated. For some experiments, kinetics was performed with blood samples recovered 3 weeks after each injection. Seven mice were used per group.

    [0748] Measurement of the Anti-S. sonnei IgG Response

    [0749] The glycoconjugate-induced anti-LPS IgG response specific for S. sonnei LPS was measured by ELISA using purified S. sonnei LPS purified from the S. sonnei reference strain (CIP 106 347) as previously described..sup.[2] Briefly, 2.5 g of purified S. sonnei LPS was coated per ELISA plate well in PBS and incubated at 4 C. overnight. After washing the wells with PBS-Tween 20 0.01%, saturation was performed by incubating the plate for 30 min at 37 C. with PBS-BSA 1%. Then, serial dilutions of mouse sera in PBS-BSA 1% were incubated for 1 h at 37 C. After washing with PBS-Tween 20 0.01%, anti-mouse IgG peroxidase-labeled conjugate (Sigma-Aldrich) was used as secondary antibody at a dilution of 1/5,000. The IgG titer was defined as the last dilution of serum giving rise to twice the OD value obtained with similarly diluted pre-immune serum. To measure the anti-S. sonnei LPS IgG subclasses, a similar ELISA was performed except that anti-mouse IgG1, IgG2a, IgG2b and IgG3 peroxidase-labeled conjugates (Sigma-Aldrich) were used as secondary antibody at a dilution of 1/5,000.

    [0750] In addition to the original TT-conjugates featuring an (AB).sub.n hapten (n=1-4), it is noted that other TT-conjugate featuring an (AB).sub.n hapten (n=4, 5) were successfully obtained by use of the same procedure.

    Example 17: Synthesis of Other (AB).SUB.n .Oligosaccharide TT-Conjugates of the InventionLinker-Equipped Oligosaccharides Featuring a 4.SUB.A.-Endchain Hydroxyl Group

    [0751] Azidopropyl Aglycon as Linker PrecursorPost-Chain Elongation Linker Introduction

    ##STR00116##

    [0752] 3-Azidopropyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (61b). Step 1: [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (29 mg, 35 mol, 0.05 equiv.) in anhyd. THF (4 mL) was stirred for 45 min under an H.sub.2 atmosphere. The pink color of the solution turned to yellow. The solution was degassed repeatedly and poured into a solution of allyl glycoside 18b (2.39 g, 693 mol, 1.0 equiv.) in anhyd. THF (20 mL). The reaction was stirred for 8 h at rt. NIS (187 mg, 831 mol, 1.2 equiv.) and H.sub.2O (5 mL) were added, and the reaction stirred for another 4 h. Following a TLC analysis, 50% aq. Na.sub.2S.sub.2O.sub.3 (10 mL) were added. The reaction mixture was concentrated under reduced pressure, and the aq. phase was diluted with DCM (50 mL) and water (50 mL). The DCM layer was washed with 50% aq. NaHCO.sub.3 (50 mL) and brine (50 mL), dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure. The crude was purified by flash chromatography (Tol/ACN, 80:20.fwdarw.75:25) to give hemiacetal 83b as a white solid (2.2 g, 645 mol, 93%). Hemiacetal 83b had R.sub.f 0.3, 0.4 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.131H.sub.134Cl.sub.24N.sub.22O.sub.37 1729.0814; found 1729.0814.

    [0753] Step 2: PTFACl (153 L, 968 mol, 1.5 equiv.) and Cs.sub.2CO.sub.3 (252 mg, 774 mol, 1.2 equiv.) were added to a solution of the hemiacetal 83b (2.2 g, 645 mol, 1.0 equiv.) in acetone (13 mL). The reaction mixture was stirred for 2 h at rt. Following a TLC analysis, the mixture filtered over a pad of Celite and washed with acetone (25 mL). The combined filtrates were concentrated under reduced pressure. Flash chromatography (Tol/ACN, 80:20) of the crude gave a mix of the required PTFA donor 85b and of the corresponding oxazoline as a white solid. Donor 85b had R.sub.f 0.55 (Tol/EtOAc 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.139H.sub.138Cl.sub.24F.sub.3N.sub.23O.sub.37 1810.6028; found 1810.6024. The oxazoline had HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.131H.sub.132Cl.sub.24N.sub.22O.sub.36 1720.0787; found 1720.0765.

    [0754] Step 3: To a solution of the PTFA donor 85b (2.0 g, 558 mol, 1.0 equiv.) in anhyd. DCE (10 mL) was added 3-azidopropanol (85 L, 838 mol, 1.5 equiv.) The solution was stirred with freshly activated MS 4 (1.0 g) under an argon atmosphere for 1 h. TMSOTf (7 L, 39 mol, 0.07 equiv. in anhyd. ACN (1 mL)) was added over 1 h while stirring the mixture at rt. After another 30 min at rt following a TLC analysis (Tol/ACN 4:1), Et.sub.3N (1 equiv. vs TMSOTf) was added. The suspension was filtered over a fitted funnel and washed with DCM (35 mL). The combined filtrate was concentrated and the yellow residue purified by automated flash chromatography (SiOH 25 m, Tol/ACN, 86:14). The azidopropyl glycoside 61b, obtained as a white solid (1.55 g, 443 mol, 79%), had R.sub.f 0.5 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.134H.sub.139Cl.sub.24N.sub.25O.sub.37 1770.6082; found 1770.6086.

    [0755] 3-Azidopropyl (benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-4-O-(2-naphthylmethyl)-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-(4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (87b). Step 1: [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (23 mg, 27 mol, 0.05 equiv.) in anhyd. THF (3 mL) was stirred for 30 min under an H.sub.2 atmosphere. The pink color of the solution turned to yellow. The mixture was degassed repeatedly with addition of argon. The solution transferred to a solution of allyl glycoside 20b (2.3 g, 540 mol, 1.0 equiv.) in anhyd. THF (25 mL). The reaction was stirred at rt. After 16 h, NIS (146 mg, 647 mol, 1.2 equiv.) and water (6 mL) were added. After stirring at rt for another 2 h, 50% aq. Na.sub.2S.sub.2O.sub.3 (10 mL) was added. Volatiles were removed under reduced pressure. The residue was diluted with DCM (40 mL) and water (40 mL). The organic layer was separated, washed with 50% aq. NaHCO.sub.3, and brine. The organic phase was collected, dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure. Flash chromatography (Tol/EtOAc 70:30.fwdarw.60:40) of the crude gave hemiacetal 84b as a white solid (2.0 g, 474 mol, 87%). Hemiacetal 84b had R.sub.f 0.5, 0.6 (Tol/ACN 6:4). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.161H.sub.163Cl.sub.30N.sub.27O.sub.46 2133.0925; found 2133.0909.

    [0756] Step 2: PTFACl (112 L, 710 mol, 1.5 equiv.) and Cs.sub.2CO.sub.3 (185 mg, 568 mol, 1.2 equiv.) were added to a solution of hemiacetal 84b (2.0 g, 474 mol, 1.0 equiv.) in acetone (10 mL). After stirring for 4 h at rt, the suspension was passed through a bed of Celite, and washed with acetone (210 mL). The combined filtrates were concentrated. Flash chromatography (cHex/EtOAc 60:40) of the crude gave the PTFA donor 86b as a white solid (2.0 g, 455 mol, 94%). Donor 86b had R.sub.f 0.5, 0.6 (Tol/ACN 6:4). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.169H.sub.167Cl.sub.30F.sub.3N.sub.28O.sub.46 2219.1060; found 2219.1055.

    [0757] Step 3: A solution of PTFA donor 76.sub.B (2.0 g, 455 mol, 1.0 equiv.) and 3-azido propanol (126 L, 1.36 mmol, 3.0 equiv.) in anhyd. DCE (22 mL) was stirred with freshly activated MS 4 (1.0 g) for 1 h at rt under an argon atmosphere. TMSOTf (6 L, 32 mol, 0.07 equiv.) in anhyd. ACN (1 mL) were added over 30 min to the reaction mixture at rt. After another 1 h at rt, a TLC analysis (Tol/ACN 4:1) showed reaction completion. Et.sub.3N (1.0 equiv. vs TMSOTf) was added. The suspension was filtered over a Celite bed, washed with DCM (210 mL). The combined filtrates were concentrated under reduced pressure. Flash chromatography (SiOH 25 m, Tol/ACN 82:18) gave decasaccharide 87b as a white solid (1.6 g, 372 mol, 81%). The azidopropyl glycoside 87b had R.sub.f 0.5 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.164H.sub.168Cl.sub.30N.sub.30O.sub.46 2174.1162; found 2174.1153.

    [0758] Full Deprotection and Linker Modification with SPDP: Chemoselective Introduction of a Masked Thiol

    ##STR00117##

    [0759] 3-Aminopropyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (67b). Protocol 4. Isopropanol (60 mL) and water (30 mL) were added to a solution of octasaccharide 61b (100 mg, 29 mol, 1.0 equiv.) in 2-MeTHF (4 mL). Sodium citrate monobasic (147 mg, 687 mol, 24 equiv.) in water (2 mL) was added followed by addition of 20% Pd(OH).sub.2/C (320 mg, 458 mol, 16 equiv.). The suspension was stirred under hydrogen for 18 h. NaHCO.sub.3 (57 mg, 687 mol, 24 equiv.) was added. The suspension was passed through a bed of Celite, and washed with methanol (210 mL), and water (210 mL). Volatiles were removed under reduced pressure. The resulting solution (50 mL) was passed through a 0.2 m syringe filter. Following lyophilization, the crude was purified by Sephadex G-10 gel filtration (100% Milli-Q water). Fractions corresponding to the desired product were lyophilized. Lastly, the residue was purified by semi-preparative RP-HPLC to give the desired octasaccharide 67b (8.6 mg, 5.1 mol, 17%) as a lyophilized white powder. Analytical data were as above.

    [0760] 3-Aminopropyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (88b). Protocol 4. Isopropanol (120 mL) and water (40 mL) were added to decasaccharide 87b (200 mg, 46 mol, 1.0 equiv.) in 2-MeTHF (8 mL), followed by sodium citrate monobasic (298 mg, 1.39 mmol, 30 equiv.) and 20% Pd(OH).sub.2/C (650 mg, 929 mol, 20 equiv.). The suspension was stirred under a hydrogen atmosphere for 24 h at rt. NaHCO.sub.3 (117 mg, 1.39 mmol, 30.0 equiv.) was added and the suspension was filtered over a bed of Celite, washed with methanol (215 mL) and then water (215 mL). The combined filtrates were concentrated (60 mL) under reduced pressure. The solution was passed through a 0.2 m syringe filter and lyophilized. The residue was purified by Sephadex G-10 gel filtration (conditions) and fractions corresponding to desired product were lyophilized. Semi-preparative RP-HPLC to give the decasaccharide 88b as a white solid (8.6 mg, 4.1 mol, 9%). The aminopropyl glycoside 88b had RP-HPLC (215 nm/ELSD): R.sub.t=8.3/8.4 min (conditions A). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.83H.sub.136N.sub.16O.sub.46 1046.4392; found 1046.4382. HRMS (ESI.sup.+): m/z [M+3H].sup.3+ calcd for C.sub.83H.sub.137N.sub.16O.sub.46 697.9619; found 697.9611.

    [0761] 3-(3-(2-Pyridyldithio)propionamido)-propyl (2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (89b). Phosphate buffer (0.4 mL, 0.2 M, pH 6.6) was added to a solution of the free decasaccharide 88b (3.0 mg, 1.4 mol, 1.0 equiv.) in Milli-Q water (0.3 mL). The solution was cooled to 5 C. A solution of SPDP (1.8 mg, 5.7 mol, 4.0 equiv.) in DMSO (15 L) was added in three portions (.sup.rd of the total volume each). After adding the first portion to the cooled, stirring was pursued for 2 h while the reaction mixture reached rt. The second portion of SDDP solution (5.0 L) was added. After stirring for another 5 h at rt, the remaining SPDP solution was added and the reaction was left overnight at rt. The reaction mixture was filtered through a 0.2 m centrifuge filter and the filtrate was subjected to semi-preparative RP-HPLC. The target decasaccharide 89b (1.2 mg, 525 nmol, 36%) was isolated as a white solid. The linker equipped decasaccharide 89b had RP-HPLC (215 nm/ELSD): R.sub.t=11.1/11.2 min (conditions D). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.91H.sub.143N.sub.17O.sub.47S.sub.2 1144.9376; found 1144.9365. HRMS (ESI.sup.+): m/z [M+3H].sup.3+ calcd for C.sub.91H.sub.144N.sub.17O.sub.47S.sub.2 763.6275; found 763.6263. HRMS (ESI.sup.+): m/z [M+H+Na].sup.2+ calcd for C.sub.91H.sub.142N.sub.17O.sub.47S.sub.2Na, 1155.9268; found 1155.9278.

    [0762] Conversion of Decasaccharide 89b into an (AB).sub.5-TT Conjugate.

    ##STR00118##

    [0763] Decasaccharide-TT conjugates (82b) from thiol-equipped decasaccharide 90b. SEC-purified tetanus toxoid (TT, 150 kD, 389 L, 6.17 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30 kD centrifugal filter four times and finally concentrated to 230 L (final concentration of TT: 8.77 mg/mL). A solution of GMBS (599 g, 2.13 mol, 160 equiv.) in DMSO (30 L) was added to the obtained solution of TT (58 M, 13 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.3 containing 5 mM EDTA, and finally concentrated to 280 L to reach a final concentration of intermediate 77b of 7.04 mg/mL.

    [0764] Decasaccharide 89b (900 g, 393 nmol) was dissolved in 0.1 M phosphate buffer pH 6.1 containing 5 mM EDTA (150 L). A 77 mM solution of tris(2-carboxyethyl)phosphine hydrochloride (TCEP.Math.HCl) in DMSO (5.4 L, 118 g, 410 nmol, 1.04 equiv.) was added to reach a concentration of 2.6 mM of oligosaccharide and the solution was stirred at rt. After 1 h, monitoring by RP-HPLC (conditions D) and HRMS revealing the absence of the starting 89b (R.sub.t=11.2 min) and the presence of a major product corresponding to the expected thiol 90b. The later had RP-HPLC (215 nm): R.sub.t=9.4 min (conditions D).

    [0765] The obtained solutions of intermediates 77b (TT, 280 L, 1.97 mg, 13.1 nmol) and crude 90b (from decasaccharide 89b, 393 nmol) were mixed and gently stirred for 4 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and adjusted to a volume of 800 L. The obtained conjugate 82b was analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0766] In addition to the above TT-conjugates featuring an (AB).sub.n hapten (n=1-5, n being the number of AB repeat per chain), it is noted that TT-conjugates featuring a B(AB).sub.n hapten ({B[AB].sub.n}m-TT, Table 2, m being the number of oligosaccharide chains per TT) have been successfully synthesized using the same procedure. To that end, the ready-for-conjugation oligosaccharides bearing a masked thiol at their reducing end were obtained. They were then conjugated analogously as described above.

    TABLE-US-00002 TABLE 2 S. sonnei B(AB).sub.n oligosaccharide-TT conjugates obtained by attachement at their non reducing end ({B[AB].sub.n}.sub.m-TT) or at their non-reducing end (TT-{B[AB].sub.n}.sub.m). Average Code in Conjugate OS:TT ratio TT conc OS conc Total FIG. 5 Nb (m) mg/mL g/mL volume Ssonnei Son O 37c1 6 2.11 48 905 L SS-TT3 Son P 37c2 13 2.26 120 950 L Son Q 37c3 20 2.25 185 860 L Ssonnei Son R 38c1 7 2.15 100 975 L SS-TT5 Son S 38c2 13 1.85 156 910 L Son T 38c3 17 1.18 138 910 L Ssonnei Son U 39c1 8 1.74 130 1520 L SS-TT7 39c2 Son V 39c3 14 0.73 96 970 L 39c4 14 0.43 56 980 L 39c5 16 0.43 65 980 L Ssonnei 40c1 11 2.24 172 800 L SS-TT5.sub.EC(End Chain) 40c2 16 1.88 210 800 L

    Example 18: Synthesis of B(AB).SUB.n .Oligosaccharides of the Invention in the Form of Azidopropyl GlycosidesPost-Chain Elongation Linker Introduction

    [0767] ##STR00119##

    [0768] 3-Azidopropyl 4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (20c). A suspension of PTFA donor 11c (2.0 g, 3.37 mmol, 1.0 equiv.) in DCM (33 mL) containing freshly activated MS 4 (1.0 g) was stirred for 1 h at rt under an argon atmosphere. After cooling to 25 C. for 10 min, TMSOTf (61 L, 337 mol, 0.1 equiv.) was added slowly. After stirring for 1 h at this temperature, a TLC analysis (Tol/EtOAc, 4:1) showed reaction completion. Et.sub.3N (1.0 equiv. vs TMSOTf) was added and the heterogenous mixture was filtered. Solids were washed with DCM (210 mL). The combined filtrates were concentrated and the crude was purified by flash chromatography (Tol/EtOAc 85:15.fwdarw.80:20) to give by order of elution the -isomer 21c (58 mg, 99 mol, 3%) and the desired -isomer 20c (1.6 g, 3.16 mmol, 82%) and, both as a white solid. The -isomer 20c had R.sub.f 0.45 (Tol/EtOAc 4:1). .sup.1H NMR (CDCl.sub.3) 7.40-7.31 (m, 5H, H.sub.Ar), 6.95 (d, 1H, J.sub.2,NH=6.0 Hz, NH), 4.91 (d, 1H, J.sub.1,2=8.4 Hz, H-1), 4.71 (d, 1H, J=11.2 Hz, CH.sub.2Bn), 4.62 (d, 1H, CH.sub.2Bn), 4.45 (dd, 1H, J.sub.2,3=10.9 Hz, J.sub.3,4=3.2 Hz, H-3), 3.96-3.91 (m.sub.po, 1H, OCH.sub.2), 3.76 (d, 1H, H-4), 3.67 (dq.sub.po, 1H, H-5), 3.62-3.55 (m.sub.po, 2H, OCH.sub.2, H-2), 3.38 (t, 2H, J=6.8 Hz, NCH.sub.2), 1.90-1.78 (m.sub.po, 2H, CH.sub.2), 1.36 (d, 3H, J.sub.5,6=6.0 Hz, H-6). .sup.13C NMR (CDCl.sub.3) 162.0 (CO.sub.NHTCA), 136.8 (C.sub.q,Ar), 128.7, 128.4, 128.3 (C.sub.Ar), 98.7 (C-1.sub.A, .sup.1J.sub.C,H=162 Hz), 92.4 (CCl.sub.3), 76.0 (C-3), 72.6 (CH.sub.2Bn), 69.1 (C-5), 66.4 (OCH.sub.2), 62.9 (C-4), 55.9 (C-2), 48.1 (NCH.sub.2), 29.0 (CH.sub.2), 17.5 (C-6). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C18H.sub.26Cl.sub.3N.sub.8O.sub.4 523.1137; found 523.1136.

    [0769] The -isomer 21c had R.sub.f 0.5 (Tol/EtOAc 4:1). .sup.1H NMR (400 MHz, CDCl.sub.3) 7.39-7.31 (m, 5H, H.sub.Ar), 6.69 (d, 1H, J.sub.2,NH=8.4 Hz, NH), 4.91 (d, 1H, J.sub.1,2=3.6 Hz, H-1), 4.76 (d, 1H, J=12.0 Hz, CH.sub.2Bn), 4.60 (d, 1H, CH.sub.2Bn), 4.45 (ddd.sub.po, 1H, H-2), 3.95 (dq.sub.po, 1H, H-5), 3.86 (dd.sub.po, 1H, J.sub.3,4=3.6 Hz, H-3), 3.83 (d.sub.po, 1H, H-4), 3.80-3.74 (m.sub.po, 1H, OCH.sub.2), 3.54-3.84 (m.sub.po, 2H, OCH.sub.2), 3.35 (t, 2H, J=5.6 Hz, NCH.sub.2), 1.88-1.84 (m.sub.po, 2H, CH.sub.2), 1.32 (d, 3H, J.sub.5,6=6.4 Hz, H-6). .sup.13C NMR (CDCl.sub.3) 161.6 (CO.sub.NHTCA), 137.8 (C.sub.q,Ar), 128.6, 128.2, 127.9 (C.sub.Ar), 96.9 (C-1, .sup.1J.sub.C,H=172 Hz), 92.6 (CCl.sub.3), 76.0 (C-3), 71.7 (CH.sub.2Bn), 65.3 (OCH.sub.2), 65.1 (C-5), 62.6 (C-4), 50.0 (C-2), 48.4 (NCH.sub.2), 28.5 (CH.sub.2), 17.4 (C-6). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.18H.sub.26Cl.sub.3N.sub.8O.sub.4 523.1137; found 523.1136.

    [0770] Allyl 4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1-4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (22c). The trisaccharide donor 14c (1.0 g, 711 mol, 1.0 equiv.) and the tetrasaccharide acceptor 15b (1.19 g, 711 mmol, 1.0 equiv.) were co-evaporated with anhyd. toluene (20 mL) and dried in high vacuum for 1 h. The dried mixture was dissolved in anhyd. DCM (18 mL) and stirred with freshly activated MS 4 (1.0 g) under an argon atmosphere for 1 h at rt. The reaction mixture was cooled to 0 C. and TMSOTf (7.7 l, 43 mol, 0.06 equiv.) was added slowly. After stirring for 1 h at 0 C., a TLC (Tol/ACN 4:1) follow up showed reaction completion. Et.sub.3N (1.0 equiv. vs TMSOTf) was added and the reaction mixture was allowed to reach rt. The suspension was filtered over a fitted funnel, washed with DCM (310 mL). The filtrate was concentrated under vacuo and the crude was purified by flash chromatography (SiOH 25 m, Tol/ACN 85:15) to give the heptasaccharide 22c as a white solid (1.46 g, 503 mol, 70%). The desired 22c had R.sub.f 0.35 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.108H.sub.116C.sub.121N.sub.21O.sub.31 1470.0799; found 1470.0819.

    [0771] 4-Azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy-/-D-galactopyranose (23c). [Ir(COD)(PMePh.sub.2).sub.2].sup.+PF.sub.6.sup. (19 mg, 22 mol, 0.05 equiv.) in anhyd. THF (3 mL) was stirred for 40 min under an H.sub.2 atmosphere. The deep red color of the solution slowly turned to yellow. The mixture was degassed and transferred to a solution of heptasaccharide 22c (1.3 g, 448 mmol, 1.0 equiv.) in anhyd. THF (9 mL). After stirring overnight at rt, NIS (121 mg, 538 mmol, 1.2 equiv.) and water (4 mL) were added. After another 2 h at rt, 50% aq. Na.sub.2S.sub.2O.sub.3 (5 mL) was added. Volatiles were removed under vacuum. The obtained solution was diluted with DCM (20 mL) and water (20 mL). The organic layer was separated and the aq. layer was washed with DCM (210). The organic parts were combined and washed with 20% aq. NaHCO.sub.3 (10 mL) and brine (10 mL). The organic phase was dried over Na.sub.2SO.sub.4, concentrated. Flash chromatography (Tol/EtOAc 70:30.fwdarw.60:40) gave the hemiacetal 23c as a white solid (1.12 g, 391 mol, 87%). Hemiacetal 23c had R.sub.f 0.5 (Tol/EtOAc 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.105H.sub.112Cl.sub.21N.sub.21O.sub.31 1450.5620; found 1450.5630. HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.105H.sub.112Cl.sub.21N.sub.21O.sub.31 1450.5620; found 1450.5630.

    [0772] 4-Azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy-/-D-galactopyranosyl (N-phenyl)trifluoroacetimidate (24c). To a solution of hemiacetal 23c (1.12 g, 391 mol, 1.0 equiv.) in acetone (15 mL) were successively added PTFA-Cl (79 L, 500 mol, 1.3 equiv.) and Cs.sub.2CO.sub.3 (150 mg, 461 mol, 1.2 equiv.). After stirring at rt for 2 h, the suspension was filtered over a Celite bed, washed with acetone (25 mL). The filtrate was concentrated. Flash chromatography (cHex/EtOAc 60:40.fwdarw.50:50) gave the PTFA donor 24c (1.09 g, 359 mol, 93%) as a white solid. The obtained 24c had R.sub.f 0.55 (Tol/EtOAc 4:1). HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.113H.sub.112Cl.sub.21F.sub.3N.sub.21O.sub.31 3053.1244; found 3053.1379.

    [0773] 3-Azidopropyl 4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-3-O-benzyl-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranosyl-(1.fwdarw.4)-(benzyl 3-O-benzyl-2-deoxy-2-trichloroacetamido--L-altropyranosyluronate)-(1.fwdarw.3)-4-azido-2-trichloroacetamido-2,4,6-trideoxy--D-galactopyranoside (25c). To a solution of the PTFA donor 24c (1.09 g, 359 mol, 1.0 equiv.) in anhyd. DCE (18 mL) was added 3-azidopropanol (66 L, 719 mol, 2.0 equiv.). The solution was stirred with freshly activated MS 4 (800 mg) for 1 h under an argon atmosphere and then cooled to 15 C. TMSOTf (7.0 L, 36 mol, 0.1 equiv., in anhyd. ACN (500 L)) was added slowly. After stirring at 15 C. for 45 min, a TLC analysis (Tol/EtOAc 7:3) showed reaction completion. Et.sub.3N (10 L) was added. The suspension was filtered over a fitted funnel. Solids were washed with DCM (25 mL) and the filtrate was concentrated. Flash chromatography (SiOH 25 m, Tol/ACN 85:15) gave the azidopropyl glycoside 25c as a white solid (750 mg, 254 mol, 70%). Heptasaccharide 25c had R.sub.f 0.3 (Tol/ACN 4:1). HRMS (ESI.sup.+): m/z [M+2NH.sub.4].sup.2+ calcd for C.sub.108H.sub.117Cl.sub.21N.sub.24O.sub.31 1492.5872; found 1492.5849. HRMS (ESI.sup.+): m/z [M+NH.sub.4].sup.+ calcd for C.sub.108H.sub.117.sup.35Cl.sub.17.sup.37Cl.sub.4N.sub.24O.sub.31 2971.1342; found 2971.1380.

    Example 19: Synthesis of Oligosaccharides of the Invention Featuring a B Endchain Residue in the Form of Aminopropyl Glycosides and of the Corresponding Ready-for-Conjugation Oligosaccharides

    [0774] ##STR00120##

    [0775] 3-Aminopropyl 2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (26c). Isopropanol (30 mL) and water (15 mL) were added to a solution of the AAT derivative 20c (50 mg, 99 mol, 1.0 equiv.) in 2-MeTHF (2 mL). NaHCO.sub.3 (25 mg, 1.18 mmol, 3.0 equiv.) in water (1 mL) was added to the solution followed by the addition of 20% Pd(OH).sub.2/C (280 mg, 396 mol, 4.0 equiv.). The suspension was stirred under a hydrogen atmosphere for 8 h at rt, filtered over a bed of Celite, washed with methanol (10 mL) and water (10 mL). The filtrate was concentrated and the crude product was purified by semi-preparative RP-HPLC. The free monosaccharide 26c was obtained as a white lyophilized solid (10.5 mg, 40 mol, 40%). Diamine 26c had RP-HPLC (215 nm/ELSD) R.sub.t=3.1/3.2 min (conditions A). .sup.1H NMR (D.sub.2O) 4.48 (d, 1H, J.sub.1,2=8.4 Hz, H-1), 4.09-3.97 (m.sub.po, 3H, H-3, H-5, OCH.sub.2), 3.77-3.67 (m.sub.po, 2H, H-2, OCH.sub.2), 3.60 (brd, 1H, J.sub.3,4=4.4 hz, H-4), 3.06 (t, 2H, J=7.2 Hz, NCH.sub.2), 2.04 (s, 3H, CH.sub.3Ac), 1.96-1.90 (m, 2H, CH.sub.2), 1.26 (d, 3H, J.sub.5,6=6.8 Hz, CH.sub.3). .sup.13C NMR (D.sub.2O) 175.1 (CO.sub.Ac), 101.6 (C-1.sub.A, .sup.1J.sub.C,H=163 Hz), 68.1 (OCH.sub.2), 67.6 (C-3), 67.5 (C-5), 54.8 (C-4), 52.1 (C-2), 37.6 (NCH.sub.2), 26.7 (CH.sub.2), 22.2 (CH.sub.3Ac), 15.7 (C-6). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.11H.sub.24N.sub.3O.sub.4 262.1761; found 262.1763.

    [0776] 3-Aminopropyl (2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (27c). Isopropanol (60 mL) and Milli-Q water (12 mL) were added to a solution of the protected trisaccharide 18c (200 mg, 152 mol, 1.0 equiv.) in 2-MeTHF (2 mL). 20% Pd(OH).sub.2/C (320 mg, 455 mol, 3.0 equiv.) was added and the mixture was stirred for 20 h at rt under hydrogen. During this timeframe, NaHCO.sub.3 (115 mg, 1.36 mmol, 9.0 equiv.) in Milli-Q water (3.0 mL) was added slowly by means of an automated syringe pump. The suspension was passed through a bed of Celite and washed with isopropanol/water (1:1, v/v, 20 mL). Volatiles were eliminated under reduced pressure and the aqueous phase was lyophilized. Semi-preparative RP-HPLC of the residue gave the aminopropyl-equipped trisaccharide 27c as a white lyophilized solid (45 mg, 68 mol, 44%). The free trisaccharide 27c had RP-HPLC (215 nm/ELSD) R.sub.t=6.2/7.3 min (conditions A). .sup.1H NMR (D.sub.2O) 4.76-4.74 (po, 1H, H-1.sub.A), 4.70 (po, 1H, H-11), 4.43 (brs, 1H, H-5.sub.A), 4.39 (d, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B), 4.09-4.06 (dd.sub.po, 1H, H-3.sub.B), 4.04-3.90 ((m.sub.po, 5H, H-3.sub.B1, H-5.sub.B, H-5.sub.B1, H-4.sub.A, OCH.sub.2), 3.85-3.72 (m.sub.po, 5H, H-2.sub.B, H-2.sub.B1, H-4.sub.B, H-2.sub.A, H-3.sub.A), 3.64-3.59 (m.sub.po, 1H, OCH.sub.2) 3.52 (br, 1H, H-4.sub.B1), 2.98 (t, 2H, J=6.4 Hz, NCH.sub.2), 1.97 (s, 3H, CH.sub.3Ac), 1.95 (s, 3H, CH.sub.3Ac), 1.91 (s, 3H, CH.sub.3Ac), 1.89-1.85 (m.sub.po, 2H, CH.sub.2), 1.26 (br, 6H, CH.sub.3). .sup.13C NMR (400 MHz, D.sub.2O) 175.4, 174.4, 174.2 (CO.sub.NHAc), 171.6 (C-6.sub.A), 103.0 (C-1.sub.B1, .sup.1J.sub.C,H=163 Hz), 101.6 (C-1.sub.B, .sup.1J.sub.C,H=163 Hz), 101.2 (C-1.sub.A, .sup.1J.sub.C,H=166 Hz), 76.6 (C-4.sub.A), 76.1 (C-5.sub.A), 75.8 (C-3.sub.B), 68.2 (OCH.sub.2), 67.6 (C-3.sub.A, C-3.sub.B1), 67.6 (C-5.sub.B), 67.4 (C-5.sub.B1), 54.8 (C-4.sub.B), 54.7 (C-4.sub.B1), 52.4 (C-2.sub.B), 51.5 (C-2.sub.A), 50.8 (C-2.sub.A1), 37.6 (NCH.sub.2), 26.7 (CH.sub.2), 22.3 (CH.sub.3Ac), 22.2 (CH.sub.3,Ac), 15.7 (C-6.sub.B), 15.6 (C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.27H.sub.49N.sub.6O.sub.13 665.3352; found 665.3336. HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.27H.sub.48N.sub.6O.sub.13Na, 687.3172; found 687.3152.

    [0777] 3-Aminopropyl (2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (28c). To a solution of the pentasaccharide 19c (120 mg, 56 mol, 1.0 equiv.) in 2-MeTHF/isopropanol/water (1:15:3, v/v/v, 76 mL) were added 20% Pd(OH).sub.2/C (160 mg, 225 mol, 4.0 equiv.) and aq. NaHCO.sub.3 (400 L, from a stock solution made of NaHCO.sub.3 (71 mg, 845 mol, 15 equiv.) in Milli-Q water (1.5 mL)). The suspension was stirred in a hydrogen atmosphere for 42 h, while slowly adding the remaining aq. NaHCO.sub.3 stock solution (200 L/2 h). At completion, the suspension was passed through a bed of Celite, washed with isopropanol (210 mL) and water (210 mL). The filtrate was concentrated under reduced pressure and lyophilized. Semi-preparative RP-HPLC purification of the residue to give the aminopropyl glycoside 28c as a white solid (12.2 mg, 11.4 mol, 20%). Pentasaccharide 28c had RP-HPLC (215 nm/ELSD): R.sub.t=8.0/8.9 min (conditions A). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.43H.sub.74N.sub.9O.sub.22 1068.4943; found 1068.4946. HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.43H.sub.75N.sub.9O.sub.22 534.7508; found 534.7512.

    [0778] 3-Aminopropyl (2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (29c). To a solution of heptasaccharide 25c (120 mg, 41 mol, 1.0 equiv.) in 2-MeTHF/isopropanol/water (1:10:2, v/v/v) was added 20% Pd(OH).sub.2/C (170 mg, 245 mol, 6.0 equiv.) and aq. NaHCO.sub.3 (600 L from a stock solution made of NaHCO.sub.3 (72 mg, 856 mol, 21.0 equiv. in water (2.1 mL)). The reaction mixture was stirred under an hydrogen atmosphere for 54 h while slowly adding the remaining aq. NaHCO.sub.3 stock solution (200 L/2 h). The suspension was passed through a bed of Celite, washed with isopropanol (210 mL) and water (210 mL). The filtrate was concentrated under reduced pressure and lyophilized. Purification by semi-preparative RP-HPLC gave the aminopropyl glycoside 29c as a white solid (17.8 mg, 12 mol, 29%). Heptasaccharide 29c had RP-HPLC (215 nm/ELSD): R.sub.t=7.4/7.5 min (conditions A). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.59H.sub.100N.sub.12O.sub.31 736.3303; found 736.3286. HRMS (ESI.sup.+): m/z [M+H+Na].sup.2+ calcd for C.sub.59H.sub.99N.sub.12O.sub.31Na, 747.3213; found 747.3197.

    [0779] 3-(3-(2-Pyridyldithio)propionamido)-propyl 2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (30c). Step 1: Isopropanol (45 mL) and water (22 mL) were added to a solution of the azidopropyl AAT 20c (132 mg, 297 mol, 1.0 equiv.) in 2-MeTHF (4.0 mL). 20% Pd(OH).sub.2/C (835 mg, 1.18 mmol, 4.0 equiv.) was added followed by the addition of NaHCO.sub.3 (75 mg, 891 mol, 3.0 equiv.) in water (1.0 mL). The reaction mixture was stirred for 8 h under a hydrogen atmosphere at rt. At completion, the suspension was filtered, washed with methanol (215 mL) and concentrated. The crude 26c was subjected to the next step without further purification.

    [0780] Step 2: SPDP linker (40 mg, 130 mol, 0.5 equiv. in DMSO (50 L)) was added to a solution of crude amine 26c (291 mol theoretical, 1.0 equiv.) in 0.1 M phosphate buffer pH 6.2 (3.3 mL, 0.1 M). The resulting suspension was stirred at rt for 48 h, during which time the precipitate disappeared slowly. At completion based on RP-HPLC monitoring, the reaction mixture was passed through a 0.2 m centrifuge filter. The filtrate was purified by semi-preparative RP-HPLC to give the desired 30c as a white lyophilized solid (29 mg, 63 mol, 24%). The linker-equipped AAT 30c had RP-HPLC (215 nm/ELSD): R.sub.t=11.0/11.0 min (conditions E). .sup.1H NMR (D.sub.2O, 400 MHz) 8.51 (d, 1H, J=4.4 Hz, H.sub.Ar), 8.12 (dt.sub.po, 1H, J=6.4 Hz, H.sub.Ar), 8.01 (d, 1H, J=8.4 Hz, H.sub.Ar), 7.55 (dt.sub.po, 1H, J=5.6 Hz, H.sub.Ar), 4.45 (d.sub.po, 1H, J.sub.1,2=8. Hz, H-1), 4.05-3.96 (m.sub.po, 2H, H-3, H-5), 3.90-3.84 (m.sub.po, 1H, OCH.sub.2), 3.74-3.69 (m.sub.po, 1H, OCH.sub.2), 3.61-3.55 (m.sub.po, 2H, H-2, H-4), 3.23-3.13 (m.sub.po, 2H, NCH.sub.2), 3.09 (t, 2H, J=6.0 Hz, CH.sub.2), 2.64 (t, 2H, J=6.8 Hz, SCH.sub.2-linker), 2.01 (s, 3H, CH.sub.3Ac), 1.74-1.68 (m, 2H, COCH.sub.2-linker), 1.30 (d, 3H, J.sub.5,6=6.4 Hz, H-6). .sup.13C NMR (D.sub.2O, 400 MHz) 174.9 (CO.sub.NHAc), 173.4 (CO.sub.linker), 157.1 (C.sub.q,Ar), 145.6, 142.4, 123.6, 123.1 (C.sub.Ar), 101.6 (C-1), 68.0 (OCH.sub.2), 67.7 (C-3), 67.5 (C-5), 54.8 (C-4), 52.1 (C-2), 36.2 (NCH.sub.2), 34.5 (SCH.sub.2), 34.1 (COCH.sub.2), 28.3 (CH.sub.2Pr), 22.2 (CH.sub.3Ac), 15.6 (C-6). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.9H.sub.31N.sub.4O.sub.5S.sub.2 459.1730; found 459.1723. HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.9H.sub.30N.sub.4O.sub.5S.sub.2Na, 481.1550; found 481.1543.

    [0781] 3-(3-(2-Pyridyldithio)propionamido)-propyl (2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (31c). The aminopropyl trisaccharide 27c (10 mg, 15 mol, 1.0 equiv.) was dissolved in 0.1 M phosphate buffer pH 6.2 (1.2 mL). A solution of SPDP (4.7 mg, 15 mol, 1.0 equiv.) in DMSO (20 L) was added slowly. The reaction mixture was stirred at rt for 48 h. At completion based on RP-HPLC monitoring, the crude reaction was passed through a 0.2 m centrifuge filter. The filtrate was purified by semi-preparative RP-HPLC to give first the unreacted 27c (4.1 mg) and then the desired 31c, both as a white lyophilized solid (3.9 mg, 4.5 mol, 30%, corrected yield 51%). Amine 31c had RP-HPLC (215 nm/ELSD): R.sub.t=16.2/17.0 min (conditions D). .sup.1H NMR (D.sub.2O, 400 MHz) 8.53 (d, 1H, J=5.6 Hz, H.sub.Ar), 8.21 (dt.sub.po, 1H, J=7.6 Hz, H.sub.Ar), 8.07 (d, 1H, J=8.4 Hz, H.sub.Ar), 7.62 (dt.sub.po, 1H, J=6.0 Hz, H.sub.Ar), 4.74 (d.sub.po, 1H, J.sub.1,2=8.8 Hz, H-1.sub.A), 4.72 (po with HOD, 1H, H-1.sub.B1), 4.61 (brs, 1H, H-5.sub.A), 4.423 (d.sub.po, 1H, H-4.sub.A), 4.40 (d, 1H, J.sub.1,2=8.6 Hz, H-1.sub.B), 4.19 (dd, 1H, J.sub.2,3=11.0 Hz, J.sub.3,4=4.0 Hz, H-331), 4.05-3.97 (m.sub.po, 3H, H-31, H-5.sub.B, H-531), 3.86-3.78 (m.sub.po, 3H, H-2.sub.A, H-231, OCH.sub.2), 3.77-3.69 (m.sub.po, 3H, H-21, H-3.sub.A, H-431), 3.57-3.51 (m.sub.po, 2H, H-41, OCH.sub.2), 3.16-3.07 (m.sub.po, 2H, NCH.sub.2), 3.06 (t.sub.po, 2H, J=6.4 Hz, COCH.sub.2-linker), 2.61 (t, 2H, J=6.4 Hz, SCH.sub.2-linker), 1.98, 1.97, 1.92 (3s, 9H, CH.sub.3Ac), 1.70-1.63 (m, 2H, CH.sub.2), 1.29 (d.sub.po, 3H, J.sub.5,6=6.8 Hz, H-6.sub.B1*), 1.26 (d.sub.po, 3H, J.sub.5,6=6.8 Hz, H-6.sub.B*). .sup.13C NMR (D.sub.2O, 400 MHz) 175.4, 174.5, 174.0 (CO.sub.NHAc), 173.4 (CO.sub.linker), 172.7 (C-6.sub.A), 156.7 (C.sub.q,Ar), 144.5, 143.7, 124.5, 123.6 (C.sub.Ar), 103.0 (C-1.sub.B1, .sup.1J.sub.C,H=165 Hz), 101.6 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 101.1 (C-1.sub., .sup.1J.sub.C,H=165 Hz), 77.0 (C-4.sub.A), 76.6 (C-5.sub.A), 76.0 (C-3.sub.B1), 68.1 (OCH.sub.2), 67.8 (C-3.sub.B), 67.6 (C-5.sub.B), 67.6 (C-5.sub.B1), 67.4 (C-3.sub.A), 54.6 (C-4.sub.B, C-4.sub.B1), 52.4 (C-2.sub.B), 51.6 (C-2.sub.A), 50.9 (C-2.sub.B1), 36.3 (NCH.sub.2), 34.5 (SCH.sub.2), 34.3 (COCH.sub.2), 28.4 (CH.sub.2Pr), 22.3, 22.3 (2C, CH.sub.3Ac), 15.7, 15.6 (C-6.sub.B, C-6.sub.B1). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.35H.sub.55N.sub.7O.sub.14S.sub.2 862.3321; found 862.3300. HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.35H.sub.55N.sub.7O.sub.14S.sub.2Na, 884.3141; found 884.3118.

    [0782] 3-(3-(2-Pyridyldithio)propionamido)-propyl (2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (32c). To a solution of the aminopropyl pentasaccharide 28c (7.1 mg, 6.7 mol, 1.0 equiv.) was dissolved in 0.1 M phosphate buffer pH 6.2 (1.6 mL). A solution of SPDP (2.0 mg, 6.7 mol, 1.0 equiv.) in DMSO (50 L) was added slowly while the reaction mixture was stirred at rt. After 8 h, more SPDP (311 g, 0.15 equiv.) in DMSO (25 L) was added. The reaction run for another 16 h and the suspension was passed through a 0.2 m centrifuge filter. The filtrate was purified by semi-preparative RP-HPLC to give first the unreacted 28c (1.6 mg) followed by the expected 32c (2.1 mg, 1.6 mol, 25%, corrected yield 32%), both as a white solid. The ready-for-conjugation pentasaccharide 32c had RP-HPLC (215 nm/ELSD): R.sub.t=11.4/11.5 min (conditions D). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.51H.sub.82N.sub.10O.sub.23S.sub.2 633.2492; found 633.2480.

    [0783] 3-(3-(2-Pyridyldithio)propionamido)-propyl (2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (33c). To a solution of the aminopropyl heptasaccharide 29c (6.0 mg, 4.1 mol, 1.0 equiv.) in 0.1 M phosphate buffer pH 6.2 (1.0 mL). SPDP (1.3 mg, 4.1 mol, 1.0 equiv.) in DMSO (30 L) was added After 20 h at rt, a LCMS analysis indicated the presence of the desired product. The suspension was filtered by passing through a 0.2 m centrifuge filter. The filtrate was purified by semi-preparative RP-HPLC to give 33c (1.8 mg, 1.42 mol, 26%) as a white lyophilized solid. The ready-for-conjugation heptasaccharide 33c had RP-HPLC (215 nm/ELSD): R.sub.t=15.5/16.4 min (conditions D). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.67H.sub.107N.sub.13O.sub.32S.sub.2 834.8266; found 834.8288. HRMS (ESI.sup.+): m/z [M+H+Na].sup.2+ calcd for C.sub.67H.sub.106N.sub.13O.sub.32S.sub.2Na, 845.8197; found 845.8171.

    Example 20: Synthesis of Oligosaccharide-TT Conjugates of the Invention from Oligosaccharides Precursors Comprising a B Endchain Residue and a Masked Thiol Linker at their Reducing End

    [0784] ##STR00121##

    [0785] Trisaccharide-TT conjugates (37c) from thiol-equipped trisaccharide 34c. SEC purified tetanus toxoid (TT, 150 kD, 1.0 mL, 6.16 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30kD centrifugal filter four times and finally concentrated to 400 L (final concentration of TT: 15.6 mg/mL). A solution of GMBS (1.85 mg, 6.6 mol, 160 equiv.) in DMSO (20 L) was added in two portions to the obtained solution of TT (104 M, 41 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.1 containing 5 mM EDTA, and finally concentrated to 410 L to reach a final concentration of intermediate 77b of 15.1 mg/mL.

    [0786] Trisaccharide 31c (300 g, 348 nmol) was dissolved in 0.1 M phosphate buffer pH 6.1 containing 5 mM EDTA (150 L). A 67 mM solution of TCEP-HCl in DMSO (5.2 L, 100 g, 349 nmol, 1.0 equiv.) was added and the solution was stirred at rt for 1 h. Monitoring was achieved by RP-HPLC (conditions D) and HRMS revealing the absence of the starting 31c (R.sub.t=15.7 min) and the presence of a major product corresponding to the expected thiol 34c. The later had RP-HPLC (215 nm/ELSD): R.sub.t=11.6/12.4 min (conditions D). Thiol 34c had LC-MS (ESI.sup.+): m/z [M].sup.+ calcd for C.sub.30H.sub.52N.sub.6O.sub.14S, 753.3; found 753.2.

    [0787] A portion of the obtained solution of crude 34c (138 L, 318 mol theo, 24 equiv.) was mixed with a portion of the stock solution of modified TT (77b, 132 L, 2.0 mg, 13.3 nmol) and gently stirred for 3 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. The final conjugate 37c1 was analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0788] In another experiment, trisaccharide 31c (1.96 mg, 2.27 mol) was dissolved in 0.1 M phosphate buffer pH 6.1 containing 5 mM EDTA (200 L). A 67 M solution of TCEP-HCl in DMSO (34 L, 653 g, 2.28 mol, 1.0 equiv.) was added and the solution was stirred at rt for 1 h. Monitoring was achieved by RP-HPLC (conditions D) as above to give the expected thiol 34c.

    [0789] Different individual portions of various stock solutions of intermediates 77b and the obtained 34c, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0790] Conjugate 37c2: From the stock solution of modified TT (77b, 132 L, 2.0 mg, 13.3 nmol) and crude thiol 34c (60 L, 580 nmol, 44 equiv.).

    [0791] Conjugate 37c3: From the stock solution of modified TT (77b, 132 L, 2.0 mg, 13.3 nmol) and crude thiol 34c (95 L, 930 nmol, 70 equiv.).

    [0792] Pentasaccharide-TT conjugates (38c) from thiol-equipped pentasaccharide 35c. SEC purified tetanus toxoid (TT, 150 kD, 1.95 mL, 5.76 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30kD centrifugal filter four times and finally concentrated to 760 L (final concentration of TT: 14.7 mg/mL). A solution of GMBS (3.3 mg, 11.8 mol, 160 equiv.) in DMSO (20 L) was added in two portions to the obtained solution of TT (97 M, 74 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.1 containing 5 mM EDTA, and finally concentrated to 650 L to reach a final concentration of intermediate of 17.15 mg/mL.

    [0793] Pentasaccharide 32c (1.8 mg, 1.4 mol) was dissolved in 0.1 M phosphate buffer pH 6.1 (150 L). A 63 mM solution of TCEP-HCl in 0.1 M phosphate buffer pH 6.1 (23 L, 410 g, 1.4 mol, 1.0 equiv.) was added and the solution was stirred at rt for 1 h. Monitoring was achieved by RP-HPLC (conditions D) and HRMS revealing the absence of the starting 32c (R.sub.t=11.2/11.4 min) and the presence of a major product corresponding to the expected thiol 35c. The later had RP-HPLC (215 nm): R.sub.t=8.8/8.9 min (conditions D). Thiol 35c had HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.46H.sub.79N.sub.9O.sub.23S m/z 578.7499; found 578.7491.

    [0794] Different individual portions of various stock solutions of intermediates 77b and 35c, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3 h at rt. Cysteamine.Math.HCl (0.4 mg, 3.0 mol, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0795] Conjugate 38c1: From the stock solution of modified TT (77b, 115 L, 2.0 mg, 13.3 nmol) and crude thiol 35c (38.7 L, 319 nmol, 24 equiv.).

    [0796] Conjugate 38c2: From the stock solution of modified TT (77b, 105 L, 1.8 mg, 12.0 nmol) and crude thiol 35c (64 L, 526 nmol, 44 equiv.).

    [0797] Conjugate 38c3: From the stock solution of modified TT (77b, 69 L, 1.2 mg, 8.0 nmol) and crude thiol 35c (67 L, 550 nmol, 70 equiv.).

    [0798] Heptasaccharide-TT conjugates (39c) from thiol-equipped heptasaccharide 36c. SEC purified tetanus toxoid (TT, 150 kD, 730 L, 6.15 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30kD centrifugal filter four times and finally concentrated to 310 L (final concentration of TT: 14.4 mg/mL). A solution of GMBS (1.3 mg, 4.7 mol, 160 equiv.) in DMSO (10 L) was added to the obtained solution of TT (96 M, 30 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.1 containing 5 mM EDTA, and finally concentrated to 310 L to reach a final concentration of intermediate 77b of 14.3 mg/mL.

    [0799] Heptasaccharide 33c (1.0 mg, 600 nmol) was dissolved in 0.1 M phosphate buffer pH 6.1 containing 5 mM EDTA (150 L). A 58 mM solution of TCEP.Math.HCl in 0.1 M phosphate buffer pH 6.1 (10.3 L, 172 g, 600 nmol, 1.0 equiv.) was added and the solution was stirred at rt for 1 h. Monitoring was achieved by RP-HPLC (conditions D) and HRMS revealing the absence of the starting 33c (R.sub.t=15.5/16.5 min) and the presence of a major product corresponding to the expected thiol 36c. The later had RP-HPLC (215 nm/ELSD): R.sub.t=11.8/12.6 min (conditions D). Thiol 36c had LC-MS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.62H.sub.105N.sub.12O.sub.32S, 780.8; found 780.5.

    [0800] Different individual portions of various stock solutions of intermediates 77b and 36c, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested.

    [0801] Conjugate 39c1: From the stock solution of modified TT (77b, 112 L, 1.6 mg, 10.6 nmol) and crude thiol 36c (68 L, 255 nmol, 24 equiv.).

    [0802] Conjugate 39c2: From the stock solution of modified TT (77b, 77 L, 1.1 mg, 7.3 nmol) and crude thiol 36c (86 L, 322 nmol, 44 equiv.).

    [0803] The two preparations were combined and the final conjugate was analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0804] In another experiment, SEC purified tetanus toxoid (TT, 150 kD, 1.0 mL, 6.17 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30kD centrifugal filter four times and finally concentrated to 420 L (final concentration of TT: 14.7 mg/mL). A solution of GMBS (1.8 mg, 6.5 mol, 160 equiv.) in DMSO (20 L) was added in two portions to the obtained solution of TT (98 M, 76 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.1 containing 5 mM EDTA, and finally concentrated to 450 L to reach a final concentration of intermediate 77b of 14.6 mg/mL.

    [0805] Heptasaccharide 33c (1.7 mg, 1.0 mol) was dissolved in 0.1 M phosphate buffer pH 6.1 containing 5 mM EDTA (150 L). A 81 mM solution of TCEP-HCl in 0.1 M phosphate buffer pH 6.1 (12.5 L, 292 g, 1.0 mol, 1.0 equiv.) was added and the solution was stirred at rt for 1 h. Monitoring was achieved by RP-HPLC (conditions D). Analytical data were as above.

    [0806] Different individual portions of various stock solutions of intermediates 77b and 36c, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0807] Conjugate 39c3: From the stock solution of modified TT (77b, 46 L, 675 g, 4.5 nmol) and crude thiol 36c (69.9 L, 440 nmol, 98 equiv.).

    [0808] Conjugate 39c4: From the stock solution of modified TT (77b, 31 L, 453 g, 3.0 nmol) and crude thiol 36c (25.4 L, 160 nmol, 53 equiv.).

    [0809] Conjugate 39c5: From the stock solution of modified TT (77b, 31 L, 453 g, 3.0 nmol) and crude thiol 36c (74.1 L, 467 nmol, 155 equiv.).

    [0810] Interestingly, single-site conjugation of the oligosaccharides at their non-reducing end was also successfully achieved to yield other conjugates of the invention, for example TT-B(AB).sub.2 pentasaccharide conjugates, using analogous procedures, albeit starting from oligosaccharides precursors comprising a B endchain residue and a masked thiol linker at their non-reducing end.

    Example 21: Synthesis of B(AB).SUB.n .Oligosaccharides in the Form of Propyl Glycosides and Conversion into Ready-for-Conjugation Oligosaccharides Featuring a Masked Thiol Linker at their Non-Reducing End

    [0811] ##STR00122##

    [0812] Propyl (2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (41c)..sup.[1] Isopropanol (90 mL) and water (30 mL) were added to a solution of the protected trisaccharide 12c (150 mg, 118 mol, 1.0 equiv.) in 2-MeTHF (4 mL). A solution of NaHCO.sub.3 (89 mg, 1.05 mmol, 9.0 equiv.) in water (1.0 mL) was added followed by the addition of Pd(OH).sub.2/C (495 mg, 706 mol, 6.0 equiv.). The reaction mixture stirred under a hydrogen atmosphere for 5 h at rt, at which time a LCMS follow up revealed the presence of the desired product. The suspension was passed through a bed of Celite. Solids were washed with isopropanol (210 mL) and water (210 mL). The filtrate was concentrated under reduced pressure and the residual volume was lyophilized. Semi-preparative RP-HPLC of the crude residue gave the propyl glycoside 41c as a white lyophilized solid (41 mg, 63 mol, 53%). Trisaccharide 41c had RP-HPLC (215 nm/ELSD): R.sub.t=13.3/13.4 min (conditions A). .sup.1H NMR (D.sub.2O) 4.78 (d.sub.po, 1H, J.sub.1,2=8.8 Hz, H-1.sub.A), 4.74 (d.sub.po, 1H, J.sub.1,2=8.4 Hz, H-1.sub.B1), 4.73 (bs.sub.po, 1H, H-5.sub.A), 4.47 (brs, 1H, H-4.sub.A), 4.46 (d.sub.po, 1H, J.sub.1,2=8.8 Hz, H-1.sub.B), 4.11 (dd.sub.po, 1H, J.sub.2,3=11.2 Hz, J.sub.3,4=4.0 Hz, H-3.sub.B1), 4.06 (dd.sub.po, 1H, J.sub.2,3=11.2 Hz, J.sub.3,4=4.8 Hz, H-3.sub.B), 4.05-3.98 (m.sub.po, 1H, H-5.sub.B, H-5.sub.B1), 3.88-3.74 (m.sub.po, 6H, H-2.sub.A, H-3.sub.A, H-2.sub.B, H-2.sub.B1, H-4.sub.B, OCH.sub.2Pr), 3.56 (d, 1H, H-4.sub.B1), 3.54-3.48 (m.sub.po, 1H, OCH.sub.2Pr), 2.00, 1.96 (2s, 9H, CH.sub.3Ac), 1.53-1.46 (m.sub.po, 2H, CH.sub.2Pr), 1.31 (d.sub.po, 3H, J.sub.5,6=6.8 Hz, H-6.sub.B), 1.30 (d.sub.po, 3H, J.sub.5,6=6.9 Hz, H-6.sub.B1), 0.82 (t, 3H, J=7.2 Hz, CH.sub.3Pr). .sup.13C NMR (D.sub.2O) 175.4, 174.5, 174.0 (CO.sub.NHAc, C-6.sub.A), 103.0 (C-1.sub.B1, .sup.1J.sub.C,H=165 Hz), 101.6 (C-1.sub.B, .sup.1J.sub.C,H=162 Hz), 101.1 (C-1.sub., .sup.1J.sub.C,H=167 Hz), 76.7 (C-4.sub.A), 76.1 (3C, C-3.sub.B, C-3.sub.B1, C-5a), 72.7 (OCH.sub.2Pr), 67.7, 67.5 (C-5.sub.B, C-5.sub.B1), 67.4 (C-3.sub.A), 54.8 (2C, C-4.sub.B, C-4.sub.B1), 52.3 (C-2.sub.B1), 51.5 (C-2.sub.A), 50.9 (C-2.sub.B), 22.3, 22.2 (3C, CH.sub.3Ac), 22.1 (CH.sub.2Pr), 15.6, 15.5 (C-6.sub.B, C-6.sub.B1), 9.5 (CH.sub.3Pr). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.27H.sub.48N.sub.5O.sub.13 m/z 650.3243; found 650.3238. HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.27H.sub.47N.sub.5O.sub.13Na m/z 672.3063; found 672.3057.

    [0813] Propyl (2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (42c). Isopropanol (75 mL) and water (25 mL) to a solution of the protected pentasaccharide 15c (100 mg, 48 mol, 1.0 equiv.) in 2-MeTHF (4 mL). A solution of NaHCO.sub.3 (56 mg, 718 mol, 14.0 equiv.) in water (1.4 mL) was added followed by the addition of 20% Pd(OH).sub.2/C (505 mg, 670 mol, 15.0 equiv.). The reaction mixture was stirred under hydrogen for 24 h at which time solid NaHCO.sub.3 (4.0 mg, 1.0 equiv. NaHCO.sub.3 was added. The suspension was passed through a bed of Celite and solids were washed with isopropanol (210 mL) and water (210 mL). Volatiles were removed under reduced pressure and remaining solution filtered through a 0.2 m syringe filter. The filtrated was lyophilized and the residue was purified by semi-preparative RP-HPLC to give the propyl glycoside 42c as a white lyophilized solid (27 mg, 26 mol, 53%). Pentasaccharide 42c had RP-HPLC (215 nm/ELSD): R.sub.t=13.4/13.5 min (conditions A). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.43H.sub.74N.sub.8O.sub.22 m/z 527.2453; found 527.2440. HRMS (ESI.sup.+): m/z [M+Na].sup.+ calcd for C.sub.43H.sub.72N.sub.8O.sub.22Na m/z 1075.4653; found 1075.4628.

    [0814] Propyl (2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (43c). Isopropanol (90 mL) and water (30 mL) were added to a solution of the fully protected heptasaccharide 22c (120 mg, 41 mol, 1.0 equiv.) in 2-MeTHF (4 mL). Solid NaHCO.sub.3 (73 mg, 869 mol, 21.0 equiv.) was added followed by 20% Pd(OH).sub.2/C (610 mg, 869 mol, 21.0 equiv.). The reaction mixture was stirred for 24 h under a hydrogen atmosphere at rt. At this time, a LCMS analysis showed that the desired compound was the major product. The suspension was filtered over a bed of Celite. The residue was washed with isopropanol (20 mL) and water (20 mL). The combined filtrate was concentrated (50 mL) and the remaining aqueous solution was filtered with 0.2 m syringe filter. The filtrate was lyophilized and the crude was purified by semi-preparative RP-HPLC to give the propyl glycoside 43c as a white lyophilized solid (39 mg, 27 mol, 64%). Heptasaccharide 43c had RP-HPLC (215 nm): R.sub.t=14.0 min (conditions A). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.59H.sub.97N.sub.11O.sub.31 m/z 728.8249; found 728.8234. HRMS (ESI.sup.+): m/z [M+H+Na].sup.2+ calcd for C.sub.59H.sub.98N.sub.11O.sub.31Na m/z 739.8158; found 739.8142.

    [0815] Propyl (2-acetamido-4-(3-(2-pyridyldithio)propionamido)-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-(2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranosyl)-(1.fwdarw.4)-(2-acetamido-2-deoxy--L-altropyranosyluronic acid)-(1.fwdarw.3)-2-acetamido-4-amino-2,4,6-trideoxy--D-galactopyranoside (44c). SPDP (14.2 mg, 46 mol, 4.0 equiv.) in DMSO (40 L) was added to a solution of pentasaccharide 42c (12.0 mg, 11.4 mol, 1.0 equiv.) in 0.2 M phosphate buffer pH 6.6 (1.2 mL). 0.2 M Phosphate buffer pH 6.6 (0.5 mL) was added. After another 16 h, the suspension had turned into a clear solution. At 36 h, more SPDP (7.1 mg, 23 mol, 2.0 equiv.) in DMSO (20 L) was added. 24 h later, the reaction mixture was filtered using a 0.2 m centrifuge tube. The filtrate was purified by semi-preparative RP-HPLC to give firstly the unreacted 44c (8.0 mg) and then the desired pentasaccharide 44c (2.1 mg, 1.66 mol, 15%, corr. 44%), both as a white lyophilized solid. The linker-equipped 37c had RP-HPLC (215 nm/ELSD): R.sub.t=12.0/12.1 min (conditions D). HRMS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.51H.sub.81N.sub.9O.sub.23S.sub.2 m/z 625.7438; found 625.7428. HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.51H.sub.80N.sub.9O.sub.23S.sub.2 m/z 1250.4803; found 1250.4785.

    Example 22: Synthesis of Oligosaccharide-TT Conjugates of the Invention from Oligosaccharides Precursors Comprising a B Endchain Residue and a Masked Thiol Linker at their Non-Reducing End

    [0816] ##STR00123##

    [0817] Pentasaccharide-TT conjugates (40c) from thiol-equipped pentasaccharide 44c. SEC purified tetanus toxoid (TT, 150 kD, 270 L, 12.27 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30kD centrifugal filter four times and finally concentrated to 210 L (final concentration of TT: 12.3 mg/mL). A solution of GMBS (744 g, 2.65 mol, 155 equiv.) in DMSO (10 L) was added in two portions to the obtained solution of TT (82 M, 17.2 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 77b was buffer exchanged with 0.1 M Phosphate buffer pH 6.1 containing 5 mM EDTA, and finally concentrated to 630 L to reach a final concentration of intermediate of 10.9 mg/mL.

    [0818] Pentasaccharide 44c (1.2 mg, 962 nmol) was dissolved in 0.1 M phosphate buffer pH 6.1 containing 5 mM EDTA (150 L). A 209 mM solution of TCEP-HCl in 0.1 M phosphate buffer pH 6.1 (4.5 L, 275 g, 960 nmol, 1.0 equiv.) was added and the solution was stirred at rt for 1 h. Monitoring was achieved by RP-HPLC (conditions D) and HRMS revealing the absence of the starting 44c (R.sub.t=11.2 min) and the presence of a major product corresponding to the expected thiol 45c. The later had RP-HPLC (215 nm): R.sub.t=10.1 min (conditions D). Thiol 45c had HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.46H.sub.77N.sub.9O.sub.23S m/z 1141.4817; found 1141.4815; m/z [M+Na].sup.+ calcd for C.sub.46H.sub.76N.sub.9O.sub.23SNa m/z 1163.4636; found 1163.4633. Owing to the presence of the unwanted dimer, the obtained crude was treated with additional TCEP leading to the crude 45c in a total volume of 300 L.

    [0819] Different individual portions of various stock solutions of intermediates 77b and 45c, respectively, were used to obtain conjugates with different carbohydrate:protein ratios. The expected amounts of each one of the two solutions were mixed and gently stirred for 3 h at rt. Cysteamine.Math.HCl (0.4 mg, 3.0 mol, 20 mg/mL in deionised water (20 L)) was added to all reaction mixtures, facilitating a molar excess of 160 compared to intermediate 77b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0820] Conjugate 40c1: From the stock solution of modified TT (77b, 115 L, 1.8 mg, 12.0 nmol) and crude thiol 45c (150 L, 480 nmol, 40 equiv.).

    [0821] Conjugate 40c2: From the stock solution of modified TT (77b, 105 L, 1.5 mg, 10.0 nmol) and crude thiol 45c (130 L, 416 nmol, 42 equiv.).

    [0822] Moreover, changing TT for CRM 197 successfully provided CRM 197-conjugates, for example tetrasaccharide (AB).sub.2CRM conjugates and octasaccharide (AB).sub.4CRM conjugates, using analogous procedures.

    TABLE-US-00003 TABLE 3 S. sonnei (AB).sub.n oligosaccharide (OS)-CRM 197 conjugates by means of The thiol-Maleimide conjugation chemistry. Average CRM 197 OS Code in Conjugate OS:CRM conc.sup.a conc Total FIG. 8 number ratio (m) mg/mL g/mL volume Ssonnei Son CA 92b1 13 1.9 350 550 L SS- Son DA 92b2 11 0.46 72 420 L CRM4 92b3 7 0.46 47 420 L Ssonnei Son EA 93b1 11 0.36 113 550 L SS- Son FA 93b2 11.5 0.44 143 450 L CRM8 Son GA 93b3 8 0.68 155 470 L

    Example 23: Conversion of Ready-for-Conjugation (AB).SUB.n .Oligosaccharides into (AB).SUB.n.-CRM Conjugates

    [0823] ##STR00124##

    [0824] Tetrasaccharide-CRM conjugates (92b) from thiol-equipped tetrasaccharide 74b. Recombinant CRM197 (CRM, 58,443 D, Provepharm Life Solutions (Marseille, France), 1.0 mL, 5.0 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30kD centrifugal filter four times and finally concentrated to 300 L (final concentration of CRM: 16.1 mg/mL). A solution of GMBS (2.9 mg, 10.2 mol, 120 equiv.) in DMSO (15 L) was added to the obtained solution of CRM (275 M, 83 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 91b was buffer exchanged with 0.1 M Phosphate buffer pH 6.2 containing 5 mM EDTA, and finally concentrated to 300 L to reach a final concentration of intermediate 91b of 14.25 mg/mL.

    [0825] Tetrasaccharide 74b was obtained from 70b (700 g, 649 nmol) as described above.

    [0826] A portion of the stock solution of modified CRM (91b, 132 L, 1.89 mg, 32 nmol) and the total volume of crude tetrasaccharide 74b (649 nmol theo., 20 equiv.) in phosphate buffer pH 6.2 containing 5 mM EDTA was mixed and gently stirred for 20 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added facilitating a molar excess of 120 compared to intermediate 91b. After stirring for 30 min at rt, the entire volume of conjugate was purified by SEC eluting with PBS, 0.5 mL/min from an Akta Superdex 200 Increase 10 300 GL column to give three fractions listed as 92b1, 92b2, and 92b3, respectively. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0827] Octasaccharide-CRM conjugates (93b) from thiol-equipped octasaccharide 76b. Recombinant CRM197 (CRM, 58,443 D, 300 L, 5.0 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.5 by passing through a 30kD centrifugal filter four times and finally concentrated to 300 L (final concentration of CRM: 4.8 mg/mL). A solution of GMBS (863 g mg, 3.1 mol, 120 equiv.) in DMSO (5.2 L) was added to the obtained solution of CRM (82 M, 25 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 91b was buffer exchanged with 0.1 M Phosphate buffer pH 6.2 containing 5 mM EDTA, and finally concentrated to 260 L to reach a final concentration of intermediate 91b of 5.1 mg/mL.

    [0828] Octasaccharide 76b was obtained from 72b (1.5 mg, 796 nmol) as described above.

    [0829] A portion of the stock solution of modified CRM (91b, 196 L, 1.0 mg, 17 nmol) and the total volume of crude octasaccharide 74b (780 nmol theo., 46 equiv.) in phosphate buffer pH 6.2 containing 5 mM EDTA was mixed and gently stirred for 3h45 at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added facilitating a molar excess of 120 compared to intermediate 91b. After stirring for 30 min at rt, the entire volume of conjugate was purified by SEC eluting with PBS, 0.5 mL/min from an Akta Superdex 200 Increase 10 300 GL column to give three fractions listed as 93b1, 93b2, and 93b3, respectively. The final conjugates were analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0830] It is also noted that the Thiol Maleimide chemistry could be successfully replaced by the Thiol-Bromoacetyl chemistry to yield other conjugates of the invention. Indeed, tetanus toxoid modified with maleimide moieties (77b) has been successfully replaced by tetanus toxoid modified with bromoacetyl groups (94b) and CRM 197 modified with maleimide moieties (91b) has been successfully replaced by CRM 197 modified with bromoacetyl moieties (95b).

    TABLE-US-00004 TABLE 4 S. sonnei (AB).sub.n oligosaccharide-TT and (AB).sub.n oligosaccharide- TT conjugates obtained by means of the thiol-bromoacetyl chemistry. Code Average TT OS in Conjugate OS:TT conc conc Total figure Nb ratio (m) mg/mL g/mL volume Ssonnei 96b1 10 2.13.sup.a 118 880 L SS-TT.sub.Ac4 Ssonnei 97b1 4.4 0.78 25 650 L SS-CRM.sub.Ac2 Ssonnei 98b1 5.5 1.47 116 760 L SS-CRM.sub.Ac4 .sup.aThe protein concentration was measured by use of the BCA assay.

    Example 24: Conjugation of Oligosaccharide Haptens onto a Carrier by Means of the Thiol-Bromoacetyl Conjugation Chemistry

    [0831] ##STR00125##

    [0832] Tetrasaccharide-TT conjugates (96b) from thiol-equipped tetrasaccharide 74b. SEC purified tetanus toxoid (TT, 150 kD, 500 L, 7.97 mg/mL) was buffer exchanged with 0.1 M HEPES pH 7.4 by passing through a 30kD centrifugal filter four times and finally concentrated to 500 L (final concentration of TT: 7.66 mg/mL). A solution of SBAP (1.5 mg, 4.9 mol, 190 equiv.) in DMSO (25 L) was added to the obtained solution of TT (51 M, 25 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 94b was buffer exchanged with 0.1 M Phosphate buffer pH 8.0 containing 5 mM EDTA, and finally concentrated to 450 L to reach a final concentration of intermediate 94b of 7.93 mg/mL.

    [0833] Tetrasaccharide 70b (2.5 mg, 2.31 mol) was dissolved in 0.1 M phosphate buffer pH 8.0 (200 L). A 145 mM solution of TCEP-HCl in DMSO (16 L, 662 g, 2.31 mol, 1.0 equiv.) was added and the solution was stirred at rt for 1 h30. Monitoring was achieved by RP-HPLC as above.

    [0834] A portion of the crude solution of tetrasaccharide 74b (74 L, 798 nmol, 60 equiv. theo) was added to a solution of crude 94b (252 L, 2.0 mg, 13 nmol) and the solution was gently stirred for 20 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added, facilitating a molar excess of 190 compared to intermediate 94b. After stirring for 30 min at rt, the entire volume of conjugate was buffer exchanged with PBS 1 pH 7.2 and finally harvested. The final conjugate (96b1) was analyzed by UV (=280 nm), BCA and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0835] Disaccharide-CRM conjugates (97b) from thiol-equipped disaccharide 73b. Recombinant CRM197 (CRM, 58,443 D, 500 L, 5.0 mg/mL) was buffer exchanged with 0.1 M phosphate buffer pH 7.8 containing 1 mM EDTA by passing through a 30kD centrifugal filter four times and finally concentrated to 450 L (final concentration of CRM: 5.0 mg/mL). A solution of SBAP (525 g, 1.7 mol, 50 equiv.) in DMSO (8.7 L) was added to a fraction of the obtained solution of CRM (86 M, 400 L, 34 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 91b was buffer exchanged with 0.1 M Phosphate buffer pH 7.8 containing 1 mM EDTA, and finally concentrated to 300 L to reach a final concentration of intermediate 95b of 5.9 mg/mL.

    [0836] Disaccharide 73b was obtained as above starting from 69b (800 g, 1.2 mol) except that phosphate buffer pH 7.8 (200 L) was used.

    [0837] Modified CRM197 (95b, 175 L, 1.03 mg, 18 nmol) was added to the solution of crude thiol 73b (1.2 mol theo, 60 equiv. theo) and the solution was gently stirred for 20 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added facilitating a molar excess of 60 compared to intermediate 95b. After stirring for 30 min at rt, the entire volume of conjugate was purified by SEC eluting from an Akta Superdex 200 Increase 10 300 GL column with PBS1 at a 0.5 mL/min to give conjugate 97b1. The final conjugate was analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0838] Tetrasaccharide-CRM conjugates (98b) from thiol-equipped tetrasaccharide 74b. Recombinant CRM197 (CRM, 58,443 D, 800 L, 5.0 mg/mL) was buffer exchanged with 0.1 M phosphate buffer pH 7.4 containing 1 mM EDTA by passing through a 30kD centrifugal filter four times and finally concentrated to 450 L (final concentration of CRM: 8.8 mg/mL). A solution of SBAP (2.0 mg, 6.5 mol, 95 equiv.) in DMSO (8.7 L) was added to the obtained solution of CRM (150 M, 450 L, 6.8 nmol). Modification was performed at ambient temperature for 1 h. After this time, the entire volume of intermediate 91b was buffer exchanged with 0.1 M Phosphate buffer pH 7.8 containing 1 mM EDTA, and finally concentrated to 300 L to reach a final concentration of intermediate 95b of 5.9 mg/mL.

    [0839] Tetrasaccharide 74b was obtained as above starting from 70b (800 g, 1.2 mol) except that phosphate buffer pH 7.8 (200 L) was used.

    [0840] Modified CRM197 (95b, 175 L, 1.03 mg, 18 nmol) of the obtained 150 M solution was added to the solution of crude thiol 74b (1.2 mol theo, 60 equiv. theo) and the solution was gently stirred for 20 h at rt. A solution of Cysteamine.Math.HCl (0.4 mg, 20 mg/mL in deionised water (20 L)) was added facilitating a molar excess of 60 compared to intermediate 95b. After stirring for 30 min at rt, the entire volume of conjugate was purified by SEC eluting from an Akta Superdex 200 Increase 10 300 GL column with PBS1 at a 0.5 mL/min to give conjugate 98b1. The final conjugate was analyzed by UV (=280 nm) and MALDI-MS for conjugation yield and carbohydrate:protein ratio.

    [0841] In addition, aminopropyl glycosides could be successfully converted to oligosaccharides equipped at their reducing end with a linker featuring a single propargyl moiety to yield ready-for-conjugation haptens compatible with conjugation chemistries other than those involving thiol precursors as exemplified for tetrasaccharide 100b.

    [0842] Alternatively, the aminopropyl glycosides could be successfully converted to oligosaccharides equipped at their reducing end with a linker featuring a single azido moiety to yield ready-for-conjugation haptens compatible with conjugation chemistries other than those involving thiol precursors as exemplified for tetrasaccharide 100b.

    Example 25. Aminopropyl Linker Modification into Conjugation-Ready Oligosaccharides

    [0843] Chemoselective Introduction of a Propargyl Moiety or an Azido Moiety

    ##STR00126##

    [0844] Ready-for-conjugation (AB).sub.2 tetrasaccharide (99b). Tetrasaccharide 65b (1.1 mg, 1.25 mol, 1.0 equiv.) was dissolved in 0.1 M phosphate buffer pH 7.8 (300 L) and stirred vigorously at rt. A solution of propargyl-N-hydroxysuccinimidyl ester (393 g, 15 mol, 1.4 equiv.) in DMSO (50 L) was added. After stirring for 30 min at rt, RP-HPLC monitoring indicated full consumption. The total volume was purified by RP-HPLC. Amide 99b (0.9 mg, 73%) was obtained as a white lyophilized powder. The linker-equipped tetrasaccharide 99b had RP-HPLC (215 nm/ELSD): R.sub.t=9.0/9.1 min (conditions D). HRMS (ESI.sup.+): m/z [M+H].sup.+ calcd for C.sub.41H.sub.66N.sub.7O.sub.21S, 992.4306, found 992.4307.

    [0845] Ready-for-conjugation (AB).sub.2 tetrasaccharide (100b). Tetrasaccharide 65b (1.4 mg, 1.6 mol, 1.0 equiv.) was dissolved in 0.1 M phosphate buffer pH 7.8 (300 L) and stirred vigorously at rt. A solution of azido-PEG2-N-hydroxysuccinimidyl ester (741 g, 2.6 mol, 1.6 equiv.) in DMSO (50 L) was added. After stirring for 3 h at rt, RP-HPLC and LC-MS monitoring indicated the presence of a novel product. The linker-equipped tetrasaccharide 100b had RP-HPLC (215 nm/ELSD): R.sub.t=9.0/9.1 min (conditions D). LC-MS (ESI.sup.+): m/z [M+2H].sup.2+ calcd for C.sub.41H.sub.70N.sub.10O.sub.22 527.23, found 527.2; [M+H].sup.+ calcd for C.sub.41H.sub.69N.sub.10O.sub.22 1053.45, found 1053.3.