Chemoenzymatic methods for synthesizing moenomycin analogs

Abstract

The present invention provides methods of synthesizing moenomycin analogs of Formula (I). The present invention also provides compositions comprising a compound of Formula (I) and kits for synthesizing compounds of Formula (I). ##STR00001##

Claims

1. A method of synthesizing a compound of Formula (I): ##STR00089## wherein R.sup.1 is C(O)NHR.sup.8, CH.sub.2OR.sup.9, or C(O)OR.sup.9; R.sup.2 and R.sup.3 are independently hydrogen, optionally substituted aliphatic, OR.sup.9, N(R.sup.8).sub.2, or C(O)NHR.sup.8; W is O or NH; R.sup.4 is hydrogen, a hydroxyl protecting group, optionally substituted aliphatic, C(O)R.sup.10, C(O)NHR.sup.8, C(NR.sup.8)NHR.sup.8, or C(O)OR.sup.9; R.sup.5 is hydrogen, an amino protecting group, optionally substituted aliphatic, or C(O)R.sup.10; R.sup.6 is hydrogen, OR.sup.9, or OR.sup.CX; wherein R.sup.CX is a carbohydrate moiety; R.sup.7 is OR.sup.9 or N(R.sup.8).sub.2; R.sup.8 is hydrogen, an amino protecting group, C(O)R.sup.10, optionally substituted aliphatic, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl; R.sup.9 is hydrogen, a hydroxyl protecting group, C(O)R.sup.10, optionally substituted aliphatic, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl; R.sup.10 is optionally substituted aliphatic, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; R.sup.11 is N(R.sup.8).sub.2; q is 0, 1, 2, 3, 4, 5, or 6; R.sup.a and R.sup.b are independently hydrogen or a hydroxyl protecting group; G is an optionally substituted C.sub.1-30 aliphatic group, wherein 0 to 10 methylene units are optionally replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene; or G is a group of Formula (a), (b), or (c): ##STR00090## wherein a is 3, 4, or 5; ##STR00091## wherein X.sub.1, X.sub.2, X.sub.3, X.sub.4, X.sub.5, X.sub.6, and X.sub.7 are each independently hydrogen or halogen; d is an integer between 1 and 25, inclusive; and e is an integer of between 2 and 25, inclusive; provided the sum of d and e is greater than 16; or ##STR00092## wherein Y is O, S, NR.sup.Y, or an optionally substituted methylene group, wherein R.sup.Y is hydrogen, optionally substituted aliphatic, or an amino protecting group; each instance of R.sup.c is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.e, SR.sup.e, NHR.sup.e, or N(R.sup.e).sub.2, wherein each instance of R.sup.e is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.e groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring; each instance of R.sup.d is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.f, SR.sup.f, NHR.sup.f, or N(R.sup.f).sub.2, wherein each instance of R.sup.f is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.f groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring; R.sup.z is hydrogen, F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.g, SR.sup.g, NHR.sup.g, or N(R.sup.g).sub.2, wherein each instance of R.sup.g is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl or two R.sup.g groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring; each instance of n is, independently, 0, 1, 2, 3, or 4; each instance of m is, independently, 0, 1, 2, 3, or 4; and x is 1, 2, 3, 4, 5, or 6; comprising: (i) providing a compound of Formula (II): ##STR00093## and (ii) reacting the compound of Formula (II) in the presence of a galactosyltransferase enzyme (GalT) with a UDP-sugar of Formula (III): ##STR00094## to yield a compound of Formula (I).

2. The method of claim 1, wherein R.sup.10 is optionally substituted aliphatic.

3. The method of claim 2, wherein R.sup.10 is methyl or CHCH.sub.2.

4. The method of claim 2, wherein R.sup.10 is substituted with halo N.sub.3, CN, NC, NCO, OCN, NCS, SCN, NO, N.sub.2, an amino group, a hydroxyl group, aryl, heteroaryl, or optionally substituted heterocyclyl.

5. The method of claim 1, wherein R.sup.10 is CH.sub.2aryl or CH.sub.2heteroaryl.

6. The method of claim 1, wherein R.sup.11 is ##STR00095##

7. The method of claim 1, wherein the compound of Formula (III) is UDP-GalNAz, UDP-galactosamine, or UDP-GalNAc.

8. The method of claim 1, wherein R.sup.1 is C(O)NH.sub.2; R.sup.2 is hydrogen; R.sup.3 is OH; W is O; R.sup.4 is hydrogen; R.sup.5 is C(O)CH.sub.3; R.sup.6 is OH; R.sup.7 is OH; R.sup.a is hydrogen; and R.sup.b is hydrogen.

9. The method of claim 1, wherein G is ##STR00096##

10. The method of claim 1, wherein R.sup.1 is C(O)NH.sub.2 or CH.sub.2OH.

11. The method of claim 1, wherein R.sup.2 is hydrogen; and R.sup.3 is OH; or R.sup.3 is hydrogen; and R.sup.2 is OH.

12. The method of claim 1, wherein WR.sup.4 is OH, NHC(O)NH.sub.2, OC(O)NH.sub.2 or NHC(NH)NH.sub.2.

13. The method of claim 1, wherein R.sup.5 is C(O)CH.sub.3.

14. The method of claim 1, wherein R.sup.6 is OH or OR.sup.CX; wherein R.sup.CX is a carbohydrate moiety.

15. The method of claim 1, wherein R.sup.7 is OH.

16. The method of claim 1, wherein the compound of Formula (II) is one of the following: ##STR00097##

17. The method of claim 1, wherein the compound of Formula (I) is one of the following: ##STR00098##

18. The method of claim 1, wherein R.sup.11 is NHC(O)R.sup.10.

19. The method of claim 18, wherein R.sup.10 is optionally substituted aliphatic.

20. The method of claim 19, wherein R.sup.10 is substituted with N.sub.3.

21. The method of claim 20, wherein R.sup.10 CH.sub.2N.sub.3.

Description

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

(1) The present invention provides methods of synthesizing moenomycin analogs and compositions and kits useful in the inventive methods. The present invention uses the enzyme GalT from for adding a sugar moiety to the polysaccharide moiety of moenomycin analogs. In certain embodiments, the present invention employs a mutant (Y289L) of the bovine enzyme GaT (Ramakrishnan et al. J. Biol. Chem. 2002, 277: 20833). The present invention is particularly useful in the preparation of moenomycin analogs where the polysaccharide moiety is modified as compared to the natural product.

(2) Moenomycin A is a natural product that inhibits peptidoglycan biosynthesis by binding to bacterial transglycosylases. Moenomycin A is a thousand times more potent than the antibiotic vancomycin, but poor pharmacokinetic properties related to the lipid side chain have prevented its use in humans. Removal of the natural lipid side chain completely abolishes biological activities. A comprehensive study of the effect of different side chains, optionally in combination with different sugar portions, on the anti-bacterial activity compared to natural moenomycin A, has been limited as most synthetic transformations employed in the removal of the natural lipid side chain and in the addition of other different side chains have also altered other structural features of the molecule. Recently, biosynthetic and semi-synthetic methodologies were disclosed which enabled SAR study of new moenomycins; e.g., see PCT Application Publication Nos. WO 2008/021367 and WO 2009/046314, incorporated herein by reference. In the '314 publication, the inventors explored groups of intermediate length and hydrophobicity, e.g., C.sub.15-farnesyl, in an effort to explore the optimal length for activity and bioavailability. The inventors have also found that groups with lengths greater than C.sub.15-farnesyl, chains substituted with halogen atoms, and chains comprising multiple aryl moieties, provide potent anti-bacterial compounds; see International PCT Application entitled Moenomycin Analogs, Methods of Synthesis, and Uses Thereof, filed on Apr. 5, 2013, which is incorporated by reference. The inventors have also designed probe compounds based on moenomycin analogs for use in screening compounds that bind to bacterial glycosyltransferases; see International PCT Application PCT/US2013/030800 entitled Methods and Compounds for Identifying Glycosyltransferase Inhibitors, filed on Mar. 13, 2013, which is incorporated by reference.

(3) Methods of Synthesis

(4) The present invention provides methods of synthesizing moenomycin analogs including a enzymatic step to attach the C ring to the E ring using GalT. In certain embodiments, the GalT is GalT Y289L. Thus in certain embodiments, the present invention provides a method of synthesizing a compound of Formula (I):

(5) ##STR00006##
wherein

(6) R.sup.1 is C(O)NHR.sup.8, CH.sub.2OR.sup.9, or C(O)OR.sup.9;

(7) R.sup.2 and R.sup.3 are independently hydrogen, optionally substituted aliphatic, OR.sup.9, N(R.sup.8).sub.2, or C(O)NHR.sup.8;

(8) W is O or NH;

(9) R.sup.4 is hydrogen, a hydroxyl protecting group, optionally substituted aliphatic, C(O)R.sup.10, C(O)NHR.sup.8, C(NR.sup.8)NHR.sup.8, or C(O)OR.sup.9;

(10) R.sup.5 is hydrogen, an amino protecting group, optionally substituted aliphatic, or C(O)R.sup.10;

(11) R.sup.6 is hydrogen, OR.sup.9, or OR.sup.CX; wherein R.sup.CX is a carbohydrate moiety;

(12) R.sup.7 is OR.sup.9 or N(R.sup.8).sub.2;

(13) R.sup.8 is hydrogen, an amino protecting group, C(O)R.sup.10, optionally substituted aliphatic, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl, or two R.sup.8 groups on the same nitrogen may be taken together to form an optionally substituted heterocyclyl;

(14) R.sup.9 is hydrogen, a hydroxyl protecting group, C(O)R.sup.10, optionally substituted aliphatic, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl;

(15) R.sup.10 is optionally substituted aliphatic, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl;

(16) R.sup.11 is optionally substituted aliphatic, (CH.sub.2).sub.qC(O)R.sup.10, N(R.sup.8).sub.2, or OR.sup.9;

(17) q is 0, 1, 2, 3, 4, 5, or 6;

(18) R.sup.a and R.sup.b are independently hydrogen or a hydroxyl protecting group;

(19) G is an optionally substituted C.sub.1-30 aliphatic group, wherein 0 to 10 methylene units are optionally replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene; or

(20) G is a group of Formula (a), (b), or (c):

(21) ##STR00007##
wherein a is 3, 4, or 5;

(22) ##STR00008##
wherein

(23) X.sub.1, X.sub.2, X.sub.3, X.sub.4, X.sub.5, X.sub.6, and X.sub.7 are each independently hydrogen or halogen;

(24) d is an integer between 1 and 25, inclusive; and

(25) e is an integer of between 2 and 25, inclusive;

(26) provided the sum of d and e is greater than 16; or

(27) ##STR00009##
wherein

(28) Y is O, S, NR.sup.Y, or an optionally substituted methylene group, wherein R.sup.Y is hydrogen, optionally substituted aliphatic, or an amino protecting group;

(29) each instance of R.sup.c is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.e, SR.sup.e, NHR.sup.e, or N(R.sup.e).sub.2, wherein each instance of R.sup.e is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.e groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring;

(30) each instance of R.sup.d is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.f, SR.sup.f, NHR.sup.f, or N(R.sup.f).sub.2, wherein each instance of R.sup.f is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.f groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring;

(31) R.sup.z is hydrogen, F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.g, SR.sup.g, NHR.sup.g, or N(R.sup.g).sub.2, wherein each instance of R.sup.g is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl or two R.sup.g groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring;

(32) each instance of n is, independently, 0, 1, 2, 3, or 4;

(33) each instance of m is, independently, 0, 1, 2, 3, or 4; and

(34) x is 1, 2, 3, 4, 5, or 6;

(35) comprising:

(36) providing a compound of formula (II):

(37) ##STR00010##
wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.a, R.sup.b, W, and G are as described herein; and

(38) reacting the compound of Formula (II) with GalT and a UDP-sugar of formula (III):

(39) ##STR00011##
wherein R.sup.11 is as described herein; to yield a compound of formula (I).

(40) As defined generally above, R.sup.1 is C(O)NHR.sup.8, CH.sub.2OR.sup.9, or C(O)OR.sup.9; wherein R.sup.8 is hydrogen, an amino protecting group, optionally substituted aliphatic, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl; and R.sup.9 is hydrogen, a hydroxyl protecting group, optionally substituted aliphatic, optionally substituted aryl, optionally substituted heterocyclyl, or optionally substituted heteroaryl. In some embodiments, R.sup.1 is C(O)NHR.sup.8. In certain embodiments, R.sup.1 is C(O)NH.sub.2. In certain embodiments, R.sup.1 is C(O)NH(alkyl). In some embodiments, R.sup.1 is CH.sub.2OR.sup.9. In certain embodiments, R.sup.1 is CH.sub.2OH. In certain embodiments, R.sup.1 is CH.sub.2O(protecting group) or CH.sub.2O(alkyl). In some embodiments, R.sup.1 is C(O)OR.sup.9. In certain embodiments, R.sup.1 is CO.sub.2H.

(41) As defined generally above, R.sup.2 is hydrogen, optionally substituted aliphatic, OR.sup.9, N(R.sup.8).sub.2, or C(O)NHR.sup.8. In some embodiments, R.sup.2 is hydrogen. In some embodiments, R.sup.2 is optionally substituted aliphatic. In certain embodiments, R.sup.2 is C.sub.1-6 alkyl. In certain embodiments, R.sup.2 is methyl. In some embodiments, R.sup.2 is OR.sup.9. In certain embodiments, R.sup.2 is OH. In certain embodiments, R.sup.2 is O(alkyl) or O(protecting group). In some embodiments, R.sup.2 is N(R.sup.8).sub.2. In certain embodiments, R.sup.2 is NH.sub.2. In certain embodiments, R.sup.2 is NH(alkyl) or NH(protecting group). In some embodiments, R.sup.2 is C(O)NHR.sup.8. In certain embodiments, R.sup.2 is C(O)NH.sub.2. In certain embodiments, R.sup.2 is C(O)NH(alkyl).

(42) As defined generally above, R.sup.3 is hydrogen, optionally substituted aliphatic, OR.sup.9, N(R.sup.8).sub.2, or C(O)NHR.sup.8. In some embodiments, R.sup.3 is hydrogen. In some embodiments, R.sup.3 is optionally substituted aliphatic. In certain embodiments, R.sup.3 is C.sub.1-6 alkyl. In certain embodiments, R.sup.3 is methyl. In some embodiments, R.sup.3 is OR.sup.9. In certain embodiments, R.sup.3 is OH. In certain embodiments, R.sup.3 is O(alkyl) or O(protecting group). In some embodiments, R.sup.3 is N(R.sup.8).sub.2. In certain embodiments, R.sup.3 is NH.sub.2. In certain embodiments, R.sup.3 is NH(alkyl) or NH(protecting group). In some embodiments, R.sup.3 is C(O)NHR.sup.8. In certain embodiments, R.sup.3 is C(O)NH.sub.2. In certain embodiments, R.sup.3 is C(O)NH(alkyl).

(43) In some embodiments, R.sup.2 is hydrogen and R.sup.3 is optionally substituted aliphatic, OR.sup.9, N(R.sup.8).sub.2, or C(O)NHR.sup.8. In some embodiments, R.sup.3 is hydrogen and R.sup.2 is optionally substituted aliphatic, OR.sup.9, N(R.sup.8).sub.2, or C(O)NHR.sup.8. In certain embodiments, R.sup.2 is hydrogen and R.sup.3 is OH. In other embodiments, R.sup.3 is hydrogen and R.sup.2 is OH.

(44) As defined generally above, W is O or NH, and R.sup.4 is hydrogen, a hydroxyl protecting group, optionally substituted aliphatic, C(O)R.sup.10, C(O)NHR.sup.8, C(NR.sup.8)NHR.sup.8, or C(O)OR.sup.9. In certain embodiments, W is O. In certain embodiments, W is NH. In some embodiments, R.sup.4 is hydrogen. In some embodiments, R.sup.4 is a hydroxyl protecting group. In some embodiments, R.sup.4 is C(O)R; wherein R.sup.10 is optionally substituted aliphatic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl. In certain embodiments, R.sup.4 is C(O)R; wherein R.sup.10 is optionally substituted alkyl. In certain embodiments, R.sup.4 is C(O)C.sub.1-6alkyl. In certain embodiments, R.sup.4 is acetyl. In some embodiments, R.sup.4 is C(O)OR.sup.9. In some embodiments, R.sup.4 is C(O)OR.sup.9; wherein R.sup.9 is aryl. In certain embodiments, R.sup.4 is C(O)OPh. In some embodiments, R.sup.4 is C(O)NHR.sup.8. In certain embodiments, R.sup.4 is C(O)NH.sub.2. In some embodiments, R.sup.4 is C(NR.sup.8)NHR.sup.8. In certain embodiments, R.sup.4 is C(NH)NH.sub.2. In certain embodiments, WR.sup.4 is OH, OC(O)NH.sub.2, NHC(O)NH.sub.2, or NHC(NH)NH.sub.2.

(45) In certain embodiments, R.sup.1 is C(O)NH.sub.2, R.sup.2 is methyl, R.sup.3 is OH, and WR.sup.4 is OC(O)NH.sub.2. In certain embodiments, R.sup.1 is C(O)NH.sub.2, R.sup.2 is hydrogen, R.sup.3 is OH, and WR.sup.4 is OC(O)NH.sub.2. In certain embodiments, R.sup.1 is C(O)NH.sub.2, R.sup.2 is OH, R.sup.3 is hydrogen, and WR.sup.4 is OH.

(46) As defined generally above, R.sup.5 is hydrogen, an amino protecting group, optionally substituted aliphatic, or C(O)R.sup.1. In some embodiments, R.sup.5 is hydrogen. In some embodiments, R.sup.5 is an amino protecting group. In some embodiments, R.sup.5 is optionally substituted aliphatic. In certain embodiments, R.sup.5 is optionally substituted alkyl. In certain embodiments, R.sup.5 is C.sub.1-6 alkyl. In some embodiments, R.sup.5 is C(O)R. In certain embodiments, R.sup.5 is C(O)R; wherein R.sup.10 is optionally substituted alkyl. In certain embodiments, R.sup.5 is C(O)C.sub.1-6alkyl. In certain embodiments, R.sup.5 is acetyl.

(47) As defined generally above, R.sup.6 is hydrogen, OR.sup.9, or OR.sup.CX; wherein R.sup.CX is a carbohydrate moiety. In some embodiments, R.sup.6 is hydrogen. In some embodiments, R.sup.6 is OR.sup.9. In certain embodiments, R.sup.6 is OH. In certain embodiments, R.sup.6 is O(protecting group). In certain embodiments, R.sup.6 is OAc. In some embodiments, R.sup.6 is OR.sup.CX. In certain embodiments, R.sup.6 is OR.sup.CX; wherein R.sup.CX is of formula:

(48) ##STR00012##
wherein R.sup.12, R.sup.13, R.sup.14, and R.sup.15 are independently hydrogen, a hydroxyl protecting group, or optionally substituted aliphatic.

(49) As defined generally above, R.sup.7 is OR.sup.9 or N(R.sup.8).sub.2. In some embodiments, R.sup.7 is OR.sup.9. In certain embodiments, R.sup.7 is OH. In certain embodiments, R.sup.7 is O(protecting group) or O(alkyl). In some embodiments, R.sup.7 is N(R.sup.8).sub.2. In certain embodiments, R.sup.7 is NH.sub.2. In certain embodiments, R.sup.7 is NH(protecting group), NH(alkyl), or N(alkyl).sub.2.

(50) In certain embodiments, R.sup.6 is OH, R.sup.7 is OH, and R.sup.5 is acetyl.

(51) As defined generally above, R.sup.11 is optionally substituted aliphatic, (CH.sub.2).sub.qC(O)R.sup.10, N(R.sup.8).sub.2, or OR.sup.9; and q is 0, 1, 2, 3, 4, 5, or 6. In certain embodiments, R.sup.11 is OH, NH.sub.2, NHC(O)C.sub.1-6 alkyl, or NHC(O)CH.sub.2N.sub.3. In certain embodiments, R.sup.11 is OH. In certain embodiments, R.sup.11 is NH.sub.2. In certain embodiments, R.sup.11 is NHC(O)C.sub.1-6 alkyl. In certain embodiments, R.sup.11 is NHC(O)CH.sub.3. In certain embodiments, R.sup.11 is NHC(O)CH.sub.2N.sub.3.

(52) In certain embodiments, R.sup.11 is optionally substituted aliphatic. In certain embodiments, R.sup.11 is optionally substituted alkyl. In certain embodiments, R.sup.11 is C.sub.1-6 alkyl. In certain embodiments, R.sup.11 is methyl. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10. In certain embodiments, R.sup.11 is CH.sub.2C(O)R.sup.10. In certain embodiments, R.sup.11 is C(O)R.sup.10. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is optionally substituted aliphatic. In certain embodiments, R.sup.11 is C(O)CH.sub.3, CH.sub.2C(O)CH.sub.3, C(O)CHCH.sub.2, or CH.sub.2C(O)CHCH.sub.2. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with halo. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with chloro, bromo, or iodo. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with fluoro. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)CH.sub.2Cl, (CH.sub.2).sub.qC(O)CH.sub.2Br, or (CH.sub.2).sub.qC(O)CH.sub.2I. In certain embodiments, R.sup.11 is C(O)CH.sub.2Cl, C(O)CH.sub.2Br, or C(O)CH.sub.2I. In certain embodiments, R.sup.11 is CH.sub.2C(O)CH.sub.2Cl, CH.sub.2C(O)CH.sub.2Br, or CH.sub.2C(O)CH.sub.2I. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)CH.sub.2CF.sub.3. In certain embodiments, R.sup.11 is C(O)CH.sub.2CF.sub.3. In certain embodiments, R.sup.11 is CH.sub.2C(O)CH.sub.2CF.sub.3. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with N.sub.3, CN, NC, NCO, OCN, NCS, SCN, NO, or N.sub.2. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)CH.sub.2N.sub.3, (CH.sub.2).sub.qC(O)CH.sub.2CN, (CH.sub.2).sub.qC(O)CH.sub.2NC, (CH.sub.2).sub.qC(O)CH.sub.2OCN, (CH.sub.2).sub.qC(O)CH.sub.2NCO, (CH.sub.2).sub.qC(O)CH.sub.2NCS, (CH.sub.2).sub.qC(O)CH.sub.2SCN, (CH.sub.2).sub.qC(O)CH.sub.2NO, or (CH.sub.2).sub.qC(O)CHN.sub.2. In certain embodiments, R.sup.11 is C(O)CH.sub.2N.sub.3, C(O)CH.sub.2CN, C(O)CH.sub.2NC, C(O)CH.sub.2OCN, C(O)CH.sub.2NCO, C(O)CH.sub.2NCS, C(O)CH.sub.2SCN, C(O)CH.sub.2NO, or C(O)CHN.sub.2. In certain embodiments, R.sup.11 is CH.sub.2C(O)CH.sub.2N.sub.3, CH.sub.2C(O)CH.sub.2CN, CH.sub.2C(O)CH.sub.2NC, CH.sub.2C(O)CH.sub.2OCN, CH.sub.2C(O)CH.sub.2NCO, CH.sub.2C(O)CH.sub.2NCS, CH.sub.2C(O)CH.sub.2SCN, CH.sub.2C(O)CH.sub.2NO, or CH.sub.2C(O)CH.sub.2N.sub.2. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with amino. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)CH.sub.2NH.sub.2. In certain embodiments, R.sup.11 is C(O)CH.sub.2NH.sub.2. In certain embodiments, R.sup.11 is CH.sub.2C(O)CH.sub.2NH.sub.2. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with hydroxy. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)CH.sub.2OH. In certain embodiments, R.sup.11 is C(O)CH.sub.2OH. In certain embodiments, R.sup.11 is CH.sub.2C(O)CH.sub.2OH. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with aryl or heteroaryl. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is CH.sub.2aryl or CH.sub.2heteroaryl. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with optionally substituted heterocyclyl. In certain embodiments, R.sup.11 is (CH.sub.2).sub.qC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with

(53) ##STR00013##

(54) In certain embodiments, R.sup.11 is N(R.sup.8).sub.2. In certain embodiments, R.sup.11 is NH.sub.2. In certain embodiments, R.sup.11 is NHC(O)R.sup.10. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is optionally substituted aliphatic. In certain embodiments, R.sup.11 is NHC(O)CH.sub.3, or NHC(O)CHCH.sub.2. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with halo. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with chloro, bromo, or iodo. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with fluoro. In certain embodiments, R.sup.11 is NHC(O)CH.sub.2Cl, NHC(O)CH.sub.2Br, or NHC(O)CH.sub.2I. In certain embodiments, R.sup.11 is NHC(O)CH.sub.2CF.sub.3. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with N.sub.3, CN, NC, NCO, OCN, NCS, SCN, NO, or N.sub.2. In certain embodiments, R.sup.11 is NHC(O)CH.sub.2N.sub.3, NHC(O)CH.sub.2CN, NHC(O)CH.sub.2NC, NHC(O)CH.sub.2OCN, NHC(O)CH.sub.2NCO, NHC(O)CH.sub.2NCS, NHC(O)CH.sub.2SCN, NHC(O)CH.sub.2NO, or NHC(O)CHN.sub.2. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with amino. In certain embodiments, R.sup.11 is NHC(O)CH.sub.2NH.sub.2. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with hydroxy. In certain embodiments, R.sup.11 is NHC(O)CH.sub.2OH. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with aryl or heteroaryl. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is CH.sub.2aryl or CH.sub.2heteroaryl. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with optionally substituted heterocyclyl. In certain embodiments, R.sup.11 is NHC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with

(55) ##STR00014##
In certain embodiments, R.sup.11 is

(56) ##STR00015##

(57) In certain embodiments, R.sup.11 is OR.sup.9. In certain embodiments, R.sup.11 is OH. In certain embodiments, R.sup.11 is O(protecting group). In certain embodiments, R.sup.11 is OAc. In certain embodiments, R.sup.11 is OC(O)R.sup.10. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is optionally substituted aliphatic. In certain embodiments, R.sup.11 is OC(O)CH.sub.3, or OC(O)CHCH.sub.2. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with halo. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with chloro, bromo, or iodo. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with fluoro. In certain embodiments, R.sup.11 is OC(O)CH.sub.2Cl, OC(O)CH.sub.2Br, or OC(O)CH.sub.2I. In certain embodiments, R.sup.11 is OC(O)CH.sub.2CF.sub.3. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with N.sub.3, CN, NC, NCO, OCN, NCS, SCN, NO, or N.sub.2. In certain embodiments, R.sup.11 is OC(O)CH.sub.2N.sub.3, OC(O)CH.sub.2CN, OC(O)CH.sub.2NC, OC(O)CH.sub.2OCN, OC(O)CH.sub.2NCO, OC(O)CH.sub.2NCS, OC(O)CH.sub.2SCN, OC(O)CH.sub.2NO, or OC(O)CHN.sub.2. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with amino. In certain embodiments, R.sup.11 is OC(O)CH.sub.2NH.sub.2. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with hydroxy. In certain embodiments, R.sup.11 is OC(O)CH.sub.2OH. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with aryl or heteroaryl. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is CH.sub.2aryl or CH.sub.2heteroaryl. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with optionally substituted heterocyclyl. In certain embodiments, R.sup.11 is OC(O)R.sup.10, wherein R.sup.10 is aliphatic substituted with

(58) ##STR00016##

(59) As defined generally above, R.sup.12 is hydrogen, a hydroxyl protecting group, or optionally substituted aliphatic. In some embodiments, R.sup.12 is hydrogen. In some embodiments, R.sup.12 is a hydroxyl protecting group. In some embodiments, R.sup.12 is optionally substituted aliphatic. In certain embodiments, R.sup.12 is C.sub.1-6 alkyl. In certain embodiments, R.sup.12 is methyl.

(60) As defined generally above, R.sup.13 is hydrogen, a hydroxyl protecting group, or optionally substituted aliphatic. In some embodiments, R.sup.13 is hydrogen. In some embodiments, R.sup.13 is a hydroxyl protecting group. In some embodiments, R.sup.13 is optionally substituted aliphatic. In certain embodiments, R.sup.13 is C.sub.1-6 alkyl. In certain embodiments, R.sup.13 is methyl.

(61) As defined generally above, R.sup.14 is hydrogen, a hydroxyl protecting group, or optionally substituted aliphatic. In some embodiments, R.sup.14 is hydrogen. In some embodiments, R.sup.14 is a hydroxyl protecting group. In some embodiments, R.sup.14 is optionally substituted aliphatic. In certain embodiments, R.sup.14 is C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is methyl.

(62) As defined generally above, R.sup.15 is hydrogen, a hydroxyl protecting group, or optionally substituted aliphatic. In some embodiments, R.sup.15 is hydrogen. In some embodiments, R.sup.15 is a hydroxyl protecting group. In some embodiments, R.sup.15 is optionally substituted aliphatic. In certain embodiments, R.sup.15 is C.sub.1-6 alkyl. In certain embodiments, R.sup.15 is methyl.

(63) In certain embodiments, R.sup.12, R.sup.13, R.sup.14, and R.sup.15 are all hydrogen.

(64) As defined generally above, R.sup.a is hydrogen or a hydroxyl protecting group. In certain embodiments, R.sup.a is hydrogen. In certain embodiments, R.sup.a is a hydroxyl protecting group.

(65) As defined generally above, R.sup.b is hydrogen or a hydroxyl protecting group. In certain embodiments, R.sup.b is hydrogen. In certain embodiments, R.sup.b is a hydroxyl protecting group.

(66) As defined generally above, G is an optionally substituted C.sub.1-30 aliphatic group, wherein 0 to 10 methylene units are optionally replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene; or

(67) G is a group of Formula (a), (b), or (c):

(68) ##STR00017##
wherein a, X.sub.1, X.sub.2, X.sub.3, X.sub.4, X.sub.5, X.sub.6, X.sub.7, d, e, Y, R.sup.c, R.sup.d, R.sup.z, x, m, and n are as described herein.

(69) In certain embodiments, G is an optionally substituted C.sub.1-30 aliphatic group, wherein 0 to 10 methylene units are optionally replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene. In certain embodiments, G is an optionally substituted C.sub.1-10, C.sub.5-20, C.sub.10-20, C.sub.12-18, or C.sub.15-20 aliphatic group, wherein 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 methylene units are optionally replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene.

(70) Exemplary aliphatic moieties include, but are not limited to, methyl (C.sub.1), ethyl (C.sub.2), propyl (C.sub.3), butyl (C.sub.4), pentyl (C.sub.5), hexyl (C.sub.6), heptyl (C.sub.7), octyl (C.sub.8), nonyl (C.sub.9), decyl (C.sub.10), undecyl (C.sub.11), dodecyl (C.sub.12), tridecyl (C.sub.13), tetradecyl (C.sub.14), pentadecyl (C.sub.15), hexadecyl (C.sub.16), heptadecyl (C.sub.17), octadecyl (C.sub.18), nonadecyl (C.sub.19), eicosyl (C.sub.20), and so on, up to (C.sub.30). In certain embodiments, the aliphatic moiety is a straight chain alkyl moiety, including, but not limited to, methyl (C.sub.1), ethyl (C.sub.2), n-propyl (C.sub.3), n-butyl (C.sub.4), n-pentyl (C.sub.5), n-hexyl (C.sub.6), n-heptyl (C.sub.7), n-octyl (C.sub.8), n-nonyl (C.sub.9), n-decyl (C.sub.10), n-undecyl (C.sub.11), n-dodecyl (C.sub.12), n-tridecyl (C.sub.13), n-tetradecyl (C.sub.14), n-pentadecyl (C.sub.15), n-hexadecyl (C.sub.16), n-heptadecyl (C.sub.17), n-octadecyl (C.sub.18), n-nonadecyl (C.sub.19), n-eicosyl (C.sub.20), and so on, up to (C.sub.30).

(71) Exemplary substituents include are described throughout, and include optionally substituted aliphatic (e.g., alkyl, alkenyl, alkynyl), optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl, halogen, OR.sup.v, N(R.sup.v).sub.2, SR.sup.v, NO.sub.2, NC, CN, N.sub.3, N(R.sup.v)NR.sup.v, CHO, C(O)R.sup.v, C(S)R.sup.v, C(NR.sup.v)R.sup.v, C(O)OR.sup.q, C(NR.sup.q)OR.sup.q, C(NR.sup.v)N(R.sup.v).sub.2, C(O)N(R.sup.v).sub.2, C(S)OR.sup.v, C(O)SR.sup.v, C(S)SR.sup.v, P(O)(OR.sup.v).sub.2, P(O).sub.2(OR.sup.v), S(O)(OR.sup.v), S(O).sub.2(OR.sup.v), P(O)N(R.sup.v).sub.2, P(O).sub.2N(R.sup.v).sub.2, S(O)N(R.sup.v).sub.2, or S(O).sub.2N(R.sup.v).sub.2; wherein each instance of R.sup.v is H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl or a protecting group. In certain embodiments, the G hydrocarbon chain is substituted with one or more optionally substituted aliphatic moieties, optionally substituted heteroaliphatic moieties, optionally substituted aryl moieties, optionally substituted heteroaryl moieties, halogen moieties, OR.sup.v moieties, N(R.sup.v).sub.2 moieties, or SR.sup.v moieties. In certain embodiments, the G hydrocarbon chain is substituted with one or more optionally substituted aliphatic moieties or OR.sup.v moieties. In certain embodiments, the G hydrocarbon chain is substituted with one or more optionally substituted aliphatic moieties. In certain embodiments, the G hydrocarbon chain is substituted with one or more optionally substituted C.sub.1-6 alkyl moieties. In certain embodiments, the G hydrocarbon chain is substituted with one or more CH.sub.3 moieties.

(72) In certain embodiments, G is an optionally substituted, optionally unsaturated, C.sub.5-20 hydrocarbon chain, wherein 0 to 10 methylene units are optionally replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene.

(73) In certain embodiments, G is an optionally substituted, optionally unsaturated, C.sub.10-20 hydrocarbon chain, wherein 0 to 8 methylene units are optionally replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene.

(74) In certain embodiments, G is an optionally substituted, optionally unsaturated, C10-18 hydrocarbon chain, wherein 0 to 6 methylene units are optionally replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene.

(75) In certain embodiments, G is an unsubstituted, optionally unsaturated hydrocarbon chain.

(76) In certain embodiments, G is an unsubstituted and saturated hydrocarbon chain. In certain embodiments, G is an unsubstituted hydrocarbon and saturated hydrocarbon chain wherein 0 to 10 methylene units are replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), S(O).sub.2 or NO.

(77) In certain embodiments, G is an unsubstituted and unsaturated hydrocarbon chain. In certain embodiments, G is an unsubstituted hydrocarbon and unsaturated hydrocarbon chain, wherein 1 to 10 methylene units are replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene.

(78) In certain embodiments, G is a substituted and saturated hydrocarbon chain. In certain embodiments, G is a substituted and saturated hydrocarbon chain, wherein 0 to 10 methylene units are replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), S(O).sub.2 or NO.

(79) In certain embodiments, G is a substituted, optionally unsaturated hydrocarbon chain. In certain embodiments, G is a substituted and unsaturated hydrocarbon chain. In certain embodiments, G is a substituted hydrocarbon and unsaturated hydrocarbon chain wherein 1 to 10 methylene units are replaced with O, NR.sup.x, S, C(O), C(NR.sup.x), S(O), SO.sub.2, NN, CN, NO, an optionally substituted arylene, an optionally substituted heterocyclylene, or an optionally substituted heteroarylene.

(80) In certain embodiments, G is an optionally substituted, optionally unsaturated, C.sub.10-C.sub.16 aliphatic moiety. In certain embodiments, G is an optionally substituted, C.sub.8-C.sub.16 alkyl moiety. In certain embodiments, G is an optionally substituted, C.sub.10-C.sub.14 alkyl moiety.

(81) In certain embodiments, G is:

(82) ##STR00018##

(83) In certain embodiments, G is:

(84) ##STR00019##

(85) In certain embodiments, G is fluorinated. G may be perfluorinated or partially fluorinated. In certain embodiments, all the hydrogen atoms of G are replaced with fluorine atoms. In certain embodiments, only a portion of the hydrogen atoms of G are replaced with fluorine atoms. In certain embodiments, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 80% of the hydrogen atoms are replaced with fluorine atoms. In certain embodiments, G comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or more fluorine atoms. In certain embodiments, G may include substituents that are or are not fluorinated. In certain embodiments, G is a fluorinated, optionally unsaturated, C.sub.10-C.sub.16 aliphatic moiety. In certain embodiments, G is a perfluorinated, optionally unsaturated, C.sub.10-C.sub.16 aliphatic moiety. In certain embodiments, G is a partially fluorinated, optionally unsaturated, C.sub.10-C.sub.16 aliphatic moiety. In certain embodiments, G is a fluorinated, C.sub.8-C.sub.16 alkyl moiety. In certain embodiments, G is a perfluorinated, C.sub.8-C.sub.16 alkyl moiety. In certain embodiments, G is a partially fluorinated, C.sub.8-C.sub.16 alkyl moiety. In certain embodiments, G is a fluorinated, C.sub.10-C.sub.14 alkyl moiety. In certain embodiments, G is a perfluorinated, C.sub.10-C.sub.14 alkyl moiety. In certain embodiments, G is a partially fluorinated, C.sub.10-C.sub.14 alkyl moiety.

(86) In certain embodiments, G is a perfluorinated, optionally unsaturated C.sub.10-C.sub.16 alkyl moiety. In certain embodiments, G is:

(87) ##STR00020## ##STR00021##

(88) In certain embodiments, G is:

(89) ##STR00022##

(90) In certain embodiments, G is:

(91) ##STR00023##

(92) In certain embodiments, G is a substituted or unsubstituted optionally unsaturated C.sub.2-30 hydrocarbon chain of the formulae:

(93) ##STR00024##
wherein is a single or double bond, and each instance of R.sup.e is, independently, H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl, halogen, OR.sup.v, N(R.sup.v).sub.2, SR.sup.v, NO.sub.2, NC, CN, N.sub.3, N(R.sup.v)NR.sup.v, CHO, C(O)R.sup.v, C(S)R.sup.v, C(NR.sup.v)R.sup.v, C(O)OR.sup.q, C(NR.sup.q)OR.sup.q, C(NR.sup.v)N(R.sup.v).sub.2, C(O)N(R.sup.v).sub.2, C(S)OR.sup.v, C(O)SR.sup.v, C(S)SR.sup.v, P(O)(OR.sup.v).sub.2, P(O).sub.2(OR.sup.v), S(O)(OR.sup.v), S(O).sub.2(OR.sup.v), P(O)N(R.sup.v).sub.2, P(O).sub.2N(R.sup.v).sub.2, S(O)N(R.sup.v).sub.2, or S(O).sub.2N(R.sup.v).sub.2; wherein each instance of R.sup.v is H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl or a protecting group; and each instance of u, s and t is, independently, 0, 1, 2, 3, 4, or 5.

(94) In certain embodiments, R.sup.e is, independently, H or optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl, halogen, OR.sup.v or N(R.sup.v).sub.2. In certain embodiments, R.sup.e is, independently, H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl or optionally substituted heteroaryl. In certain embodiments, R.sup.e is, independently, optionally substituted aliphatic or optionally substituted heteroaliphatic. In certain embodiments, R.sup.e is, independently, H or optionally substituted aliphatic. In certain embodiments, R.sup.e is, independently, H or CH.sub.3.

(95) In certain embodiments, G is a fully saturated hydrocarbon group of the formula:

(96) ##STR00025##

(97) wherein R.sup.e, s and t are as defined above and herein.

(98) In certain embodiments, G is a fully saturated hydrocarbon group of the formulae:

(99) ##STR00026##
wherein s and t are as defined above and herein.

(100) In certain embodiments, G is an unsaturated group of the formulae:

(101) ##STR00027##
wherein R.sup.e, s, and t are as defined herein.

(102) In certain embodiments, G is a group of the formula:

(103) ##STR00028##

(104) wherein Ring X, Ring Y and Ring Z are, independently, an optionally substituted arylene or an optionally substituted heteroarylene moiety;

(105) z is 0 to 3;

(106) each instance of j is, independently, 1 or 2; and

(107) each instance of L.sup.1 and L.sup.2 are, independently, (C(R.sup.o).sub.2, O, NR.sup.x, S, C(O), C(O)O, C(O)NR.sup.x, C(O)S, C(NR.sup.x), C(NR.sup.x)O, C(NR.sup.x)NR.sup.x, C(NR.sup.x)S, S(O), S(O).sub.2, NN, CN, C(R.sup.y)C(R.sup.y), or NO, wherein R.sup.o is H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl, each instance of R.sup.x is, independently, H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl or an amino protecting group, and each instance of R.sup.y is, independently, H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, or optionally substituted heteroaryl.

(108) Exemplary optionally substituted arylene groups include, but are not limited to:

(109) ##STR00029##

(110) wherein each instance of R.sup.s is, independently, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl, halogen, OR.sup.v, N(R.sup.v).sub.2, SR.sup.v, NO.sub.2, NC, CN, N.sub.3, N(R.sup.v)NR.sup.v, CHO, C(O)R.sup.v, C(S)R.sup.v, C(NR.sup.v)R.sup.v, C(O)OR.sup.q, C(NR.sup.q)OR.sup.q, C(NR.sup.v)N(R.sup.v).sub.2, C(O)N(R.sup.v).sub.2, C(S)OR.sup.v, C(O)SR.sup.v, C(S)SR.sup.v, P(O)(OR.sup.v).sub.2, P(O).sub.2(OR.sup.v), S(O)(OR.sup.v), S(O).sub.2(OR.sup.v), P(O)N(R.sup.v).sub.2, P(O).sub.2N(R.sup.v).sub.2, S(O)N(R.sup.v).sub.2, or S(O).sub.2N(R.sup.v).sub.2; wherein each instance of R.sup.v is H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl or a protecting group; and w is an integer between 0 to 10, inclusive.

(111) Exemplary optionally substituted heteroarylene groups include, but are not limited to:

(112) ##STR00030## ##STR00031## ##STR00032##

(113) wherein each instance of R.sup.s is, independently, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl, halogen, OR.sup.v, N(R.sup.v).sub.2, SR.sup.v, NO.sub.2, NC, CN, N.sub.3, N(R.sup.v)NR.sup.v, CHO, C(O)R.sup.v, C(S)R.sup.v, C(NR.sup.v)R.sup.v, C(O)OR.sup.q, C(NR.sup.q)OR.sup.q, C(NR.sup.v)N(R.sup.v).sub.2, C(O)N(R.sup.v).sub.2, C(S)OR.sup.v, C(O)SR.sup.v, C(S)SR.sup.v, P(O)(OR.sup.v).sub.2, P(O).sub.2(OR.sup.v), S(O)(OR.sup.v), S(O).sub.2(OR.sup.v), P(O)N(R.sup.v).sub.2, P(O).sub.2N(R.sup.v).sub.2, S(O)N(R.sup.v).sub.2, or S(O).sub.2N(R.sup.v).sub.2; wherein each instance of R.sup.v is H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl or a protecting group; and w is an integer between 0 to 10, inclusive, and each instance of R.sup.g is, independently, H, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted aryl, optionally substituted heteroaryl or an amino protecting group.

(114) In certain embodiments, Ring X, Ring Y and Ring Z are, independently, an optionally substituted arylene moiety. In certain embodiments, Ring X, Ring Y and Ring Z are, independently, an optionally substituted phenylene moiety. For example, in certain embodiments, G is an group of the formulae:

(115) ##STR00033##
wherein z, w, j, L.sup.1, L.sup.2, and R.sup.s are as defined above and herein.

(116) In certain embodiments, G is of one of the formulae:

(117) ##STR00034##

(118) In certain embodiments, G is any one of the following groups:

(119) ##STR00035## ##STR00036##

(120) In certain embodiments, G is the geranyl group:

(121) ##STR00037##

(122) In certain embodiments, G is the farnesyl group:

(123) ##STR00038##

(124) In certain embodiments, G is C.sub.12 alkyl of the formula:

(125) ##STR00039##

(126) In certain embodiments, G is the nerolyl group:

(127) ##STR00040##

(128) In certain embodiments, G is of the formula:

(129) ##STR00041##
wherein each occurrence of R.sup.e is independently hydrogen or an optionally substituted aliphatic moiety. In certain embodiments, R.sup.e is hydrogen or C.sub.1-C.sub.6 aliphatic. In certain embodiments, Re is hydrogen, C.sub.1-C.sub.6 alkyl, or C.sub.1-C.sub.6 alkenyl. In certain embodiments, R.sup.e is vinyl. In certain embodiments, R.sup.e is allyl. In certain embodiments, R.sup.e is isopropyl. In certain embodiments, R.sup.e is n-propyl. In certain embodiments, R.sup.e is isobutyl. In certain embodiments, R.sup.e is n-butyl. In certain embodiments, R.sup.e is t-butyl. In certain embodiments, R.sup.e is n-pentyl. In certain embodiments, R.sup.e is isopentyl. In certain embodiments, R.sup.e is neopentyl. In certain embodiments, R.sup.e is 3-methyl-but-2-enyl. Exemplary G groups include:

(130) ##STR00042## ##STR00043##

(131) In certain embodiments, G is of the formula:

(132) ##STR00044##

(133) In some embodiments, G is of Formula (a):

(134) ##STR00045##
wherein a is 3, 4, or 5.

(135) For example, in certain embodiments, G is:

(136) ##STR00046##
wherein a is 3;

(137) ##STR00047##
wherein a is 4; or

(138) ##STR00048##
wherein a is 5.

(139) In some embodiments, G is of Formula (b):

(140) ##STR00049##
wherein:

(141) X.sub.1, X.sub.2, X.sub.3, X.sub.4, X.sub.5, X.sub.6, and X.sub.7 are each independently hydrogen or halogen;

(142) d is an integer between 1 and 25, inclusive; and

(143) e is an integer of between 2 and 25, inclusive;

(144) provided the sum of d and e is greater than 16.

(145) In certain embodiments, e is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 24, or 25. In certain embodiments, d is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 24, or 25. Any particular combination of e or d is contemplated, provided the sum of d and e is greater than 16.

(146) For example, in certain embodiments, e is 16 or an integer greater than 16, and d is 1 or an integer greater than 1. In certain embodiments, e is 15, and d is 2 or an integer greater than 2. In certain embodiments, e is 14, and d is 3 or an integer greater than 3. In certain embodiments, e is 13, and d is 4 or an integer greater than 4. In certain embodiments, e is 12, and d is 5 or an integer greater than 5. In certain embodiments, e is 11, and d is 6 or an integer greater than 6. In certain embodiments, e is 10, and d is 7 or an integer greater than 7. In certain embodiments, e is 9, and d is 8 or an integer greater than 8. In certain embodiments, e is 8, and d is 9 or an integer greater than 9. In certain embodiments, e is 7, and d is 10 or an integer greater than 10. In certain embodiments, e is 6, and d is 11 or an integer greater than 11. In certain embodiments, e is 5, and d is 12 or an integer greater than 12. In certain embodiments, e is 4, and d is 13 or an integer greater than 13. In certain embodiments, e is 3, and d is 14 or an integer greater than 14. In certain embodiments, e is 2, and d is 15 or an integer greater than 15. In certain embodiments, e is 10, and d is 7 or an integer greater than 7, e.g., d is 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 24, or 25. In certain embodiments, e is 10 and d is 7. In certain embodiments, e is 10 and d is 8. In certain embodiments, e is 10 and d is 9. In certain embodiments, e is 10 and d is 10. In certain embodiments, e is 10 and d is 11. In certain embodiments, e is 10 and d is 12. In certain embodiments, e is 10 and d is 13. In certain embodiments, e is 10 and d is 14. In certain embodiments, e is 10 and d is 15.

(147) In certain embodiments, each instance of X.sub.1 and X.sub.2 is hydrogen. In certain embodiments, each instance of X.sub.1 and X.sub.2 is halogen, e.g., fluoro.

(148) In certain embodiments, each instance of X.sub.3 and X.sub.4 is hydrogen. In certain embodiments, each instance of X.sub.3 and X.sub.4 is halogen, e.g., fluoro.

(149) In certain embodiments, each instance of X.sub.5, X.sub.6, and X.sub.7 is hydrogen. In certain embodiments, each instance of X.sub.5, X.sub.6, and X.sub.7 is halogen, e.g., fluoro.

(150) In certain embodiments, each instance of X.sub.1, X.sub.2, X.sub.3, X.sub.4, X.sub.5, X.sub.6, and X.sub.7 is hydrogen, i.e., to provide an n-alkyl group. Exemplary n-alkyl groups of Formula (b) include, but are not limited to:

(151) ##STR00050##

(152) In certain embodiments, each instance of X.sub.1 and X.sub.2 is fluoro, optionally wherein each instance of X.sub.3 and X.sub.4 is fluoro and/or each instance of X.sub.5, X.sub.6, and X.sub.7 is fluoro. Alternatively, each instance of X.sub.3 and X.sub.4 is fluoro, optionally wherein each instance of X.sub.1 and X.sub.2 is fluoro and/or each instance of X.sub.5, X.sub.6, and X.sub.7 is fluoro. In certain embodiments, X.sub.1 and X.sub.2 are each hydrogen, X.sub.3 and X.sub.4 are each fluoro, and X.sub.5, X.sub.6, and X.sub.7 are each fluoro. In certain embodiments, X.sub.1, X.sub.2 are each fluoro, X.sub.3 and X.sub.4 are each hydrogen, and X.sub.5, X.sub.6, and X.sub.7 are each hydrogen.

(153) Exemplary fluoroalkyl groups of formula (b), wherein X.sub.1 and X.sub.2 are hydrogen and X.sub.3, X.sub.4, X.sub.5, X.sub.6, and X.sub.7 are each fluoro include, but are not limited to:

(154) ##STR00051##
wherein e is 10, and d is 7;

(155) ##STR00052##
wherein e is 10, and d is 8;

(156) ##STR00053##
wherein e is 10, and d is 9;

(157) ##STR00054##
wherein e is 10, and d is 10;

(158) ##STR00055##
wherein e is 10, and d is 11;

(159) ##STR00056##
wherein e is 10, and d is 12;

(160) ##STR00057##
wherein e is 10, and d is 13;

(161) ##STR00058##
wherein e is 10, and d is 14;

(162) ##STR00059##
wherein e is 10, and d is 15.

(163) Exemplary fluoroalkyl groups of formula (b), wherein each instance of X.sub.1, X.sub.2, X.sub.3, X.sub.4, X.sub.5, X.sub.6, and X.sub.7 is fluoro, include but are not limited to:

(164) ##STR00060##
wherein e is 10, and d is 7;

(165) ##STR00061##
wherein e is 10, and d is 8;

(166) ##STR00062##
wherein e is 10, and d is 9;

(167) ##STR00063##
wherein e is 10, and d is 10;

(168) ##STR00064##
wherein e is 10, and d is 11;

(169) ##STR00065##
wherein e is 10, and d is 12;

(170) ##STR00066##
wherein e is 10, and d is 13;

(171) ##STR00067##
wherein e is 10, and d is 14;

(172) ##STR00068##
wherein e is 10, and d is 15.

(173) In some embodiments G is of Formula (c):

(174) ##STR00069##
wherein:

(175) Y is O, S, NR.sup.Y, or an optionally substituted methylene group, wherein R.sup.Y is hydrogen, optionally substituted aliphatic, or an amino protecting group;

(176) each instance of R.sup.c is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.e, SR.sup.e, NHR.sup.e, and N(R.sup.e).sub.2, wherein each instance of R.sup.e is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.e groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring;

(177) each instance of R.sup.d is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.f, SR.sup.f, NHR.sup.f, or N(R.sup.f).sub.2, wherein each instance of R.sup.f is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.f groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring;

(178) R.sup.z is hydrogen, F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.g, SR.sup.g, NHR.sup.g, or N(R.sup.g).sub.2, wherein each instance of R.sup.g is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl or two R.sup.g groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring;

(179) each instance of n is, independently, 0, 1, 2, 3, or 4;

(180) each instance of m is, independently, 0, 1, 2, 3, or 4; and

(181) x is 1, 2, 3, 4, 5, or 6.

(182) As generally defined above, Y is O, S, NR.sup.Y, or an optionally substituted methylene group, wherein R.sup.Y is hydrogen, optionally substituted aliphatic, or an amino protecting group. In certain embodiments, Y is O. In certain embodiments, Y is S. In certain embodiments, Y is NR.sup.Y. In certain embodiments, Y is an optionally substituted methylene group, e.g., CH.sub.2.

(183) As generally defined above, each instance of R.sup.c is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.e, SR.sup.e, NHR.sup.e, or N(R.sup.e).sub.2, wherein each instance of R.sup.e is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.e groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring; and n is 0, 1, 2, 3, or 4.

(184) In certain embodiments, each instance of R.sup.c is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.e, SR.sup.e, NHR.sup.e, or N(R.sup.e).sub.2, wherein each instance of R.sup.e is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.e groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring; wherein each instance of aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl and heteroaryl is independently unsubstituted or substituted with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more substituents, as defined herein. In certain embodiments, each instance of aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl and heteroaryl is independently unsubstituted or substituted with C.sub.1-6alkyl, C.sub.1-6fluoroalkyl or halogen.

(185) In certain embodiments, each instance of R.sup.c is independently F, aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl, or heteroaryl, wherein each instance of aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl and heteroaryl is independently unsubstituted or substituted with C.sub.1-6alkyl, C.sub.1-6fluoroalkyl or halogen.

(186) In certain embodiments, each instance of R.sup.c is independently F or alkyl, wherein each instance of alkyl is independently unsubstituted or substituted with C.sub.1-6alkyl, C.sub.1-6fluoroalkyl or halogen.

(187) In certain embodiments, n is 0 or 1. In certain embodiments, n is 0. In certain embodiments, n is 1. In certain embodiments, n is 2.

(188) As generally defined above, each instance of R.sup.d is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.f, SR.sup.f, NHR.sup.f, or N(R.sup.f).sub.2, wherein each instance of R.sup.f is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.f groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring; and m is 0, 1, 2, 3, or 4.

(189) In certain embodiments, each instance of R.sup.d is independently F, Br, I, Cl, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.f, SR.sup.f, NHR.sup.f, or N(R.sup.f).sub.2, wherein each instance of R.sup.f is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl, or two R.sup.f groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring; wherein each instance of aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl and heteroaryl is independently unsubstituted or substituted with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more substituents, as defined herein. In certain embodiments, each instance of aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl and heteroaryl is independently unsubstituted or substituted with C.sub.1-6alkyl, C.sub.1-6fluoroalkyl or halogen.

(190) In certain embodiments, each instance of R.sup.d is independently F, aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl, or heteroaryl, wherein each instance of aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl, and heteroaryl is independently unsubstituted or substituted with C.sub.1-6alkyl, C.sub.1-6fluoroalkyl or halogen.

(191) In certain embodiments, each instance of R.sup.d is independently F or alkyl, wherein each instance of alkyl is independently unsubstituted or substituted with C.sub.1-6alkyl, C.sub.1-6fluoroalkyl, or halogen.

(192) In certain embodiments, m is 0 or 1. In certain embodiments, m is 0. In certain embodiments, m is 1. In certain embodiments, m is 2.

(193) In certain embodiments, R.sup.z is an ortho, meta, or para substituent to the OCH.sub.2 linking group. In certain embodiments, R.sup.z is a meta substituent.

(194) As generally defined above, R.sup.z is hydrogen, F, Br, I, Cl, optionally substituted aliphatic, heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.g, SR.sup.g, NHR.sup.g, or N(R.sup.g).sub.2, wherein each instance of R.sup.g is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl or two R.sup.g groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring.

(195) In certain embodiments, R.sup.z is hydrogen, F, Br, I, Cl, optionally substituted aliphatic, heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, optionally substituted heteroaryl, OR.sup.g, SR.sup.g, NHR.sup.g, or N(R.sup.g).sub.2, wherein each instance of R.sup.g is independently hydrogen, optionally substituted aliphatic, optionally substituted heteroaliphatic, optionally substituted carbocycyl, optionally substituted heterocycyl, optionally substituted aryl, or optionally substituted heteroaryl or two R.sup.g groups are joined to form a 5- to 6-membered optionally substituted heterocycyl or optionally substituted heteroaryl ring, wherein each instance of aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl and heteroaryl is independently unsubstituted or substituted with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more substituents, as defined herein. In certain embodiments, each instance of aliphatic, heteroaliphatic, carbocycyl, heterocycyl, aryl and heteroaryl is independently unsubstituted or substituted with C.sub.1-6alkyl, C.sub.1-6fluoroalkyl or halogen.

(196) In certain embodiments, R.sup.z is hydrogen, alkyl, alkenyl, alkynyl, carbocycyl, heterocycyl, aryl, heteroaryl, OR.sup.g, SR.sup.g, NHR.sup.g, or N(R.sup.g).sub.2, wherein each instance of R.sup.8 is independently hydrogen, alkyl, alkenyl, alkynyl, carbocycyl, heterocycyl, aryl, heteroaryl, or two R.sup.g groups are joined to form a 5- to 6-membered heterocycyl or heteroaryl ring, and wherein each instance of alkyl, alkenyl, alkynyl, carbocycyl, heterocycyl, aryl, and heteroaryl, is independently unsubstituted or substituted with C.sub.1-6alkyl, C.sub.1-6fluoroalkyl or halogen.

(197) In certain embodiments, R.sup.z is hydrogen or aryl, wherein aryl is unsubstituted or substituted with C.sub.1-6alkyl, C.sub.1-6fluoroalkyl or halogen.

(198) As generally depicted above, x is 1, 2, 3, 4, 5, or 6. In certain embodiments, x is 1 or 2. In certain embodiments, x is 1. In certain embodiments, x is 2.

(199) It will be understood by one skilled in the art that each repeat unit of formula (c), when x is greater than 1, can optionally differ from one another, arising from differences in the independent variables Y, R.sup.c, R.sup.d, n and m, as well as different substitution patterns on and between each repeating unit. Thus, in further defining the compounds of the present invention, it is also generally helpful to further designate Y, R.sup.c, R.sup.d, n and m, with a sequential number corresponding to the first, second, third, fourth, fifth or sixth sequential group from which it is formally a member, e.g., Y, R.sup.c, R.sup.d, n, m and x can also be referred to as Y.sup.1, R.sup.c1, R.sup.d1, n1 and m1 for the first group in the sequence; Y.sup.2, R.sup.c2, R.sup.d2, n2 and m2 for the second optional repeating unit in the sequence; Y.sup.3, R.sup.c3, R.sup.d3, n3 and m3 for the third optional repeating unit in the sequence; Y.sup.4, R.sup.c4, R.sup.d4, n4 and m4 for the fourth optional repeating unit in the sequence; Y.sup.5, R.sup.c5, R.sup.d5, n5 and m5 for the fifth optional repeating unit in the sequence; and Y.sup.6, R.sup.c6, R.sup.d6, n6 and m6 for the sixth optional repeating unit in the sequence.

(200) For example, in certain embodiments, the group of Formula (c) is of the formula:

(201) ##STR00070##
wherein x is 1;

(202) ##STR00071##
wherein x is 2;

(203) ##STR00072##
wherein x is 3;

(204) ##STR00073##
wherein x is 4;

(205) ##STR00074##
wherein x is 5;
or

(206) ##STR00075##
wherein x is 6;
wherein:

(207) R.sup.c1, R.sup.c2, R.sup.c3, R.sup.c4, R.sup.c5, and R.sup.c6 each independently correspond to the definition and various embodiments of R.sup.c;

(208) R.sup.d1, R.sup.d2, R.sup.d3, R.sup.d4, R.sup.d5, and R.sup.d6 each independently correspond to the definition and various embodiments of R.sup.d;

(209) n1, n2, n3, n4, n5, and n6 each independently correspond to the definition and various embodiments of n;

(210) m1, m2, m3, m4, m5, and m6 each independently correspond to the definition and various embodiments of m;

(211) Y.sup.1, Y.sup.2, Y.sup.3, Y.sup.4, Y.sup.5, and Y.sup.6, each independently correspond to the definition and various embodiments of Y;

(212) and R.sup.z is as defined herein.

(213) In certain embodiments, the group of Formula (c) is of the formula:

(214) ##STR00076##
wherein Y, R.sup.z, R.sup.c, R.sup.d, m, n, and x are as defined herein.

(215) In certain embodiments, the group of Formula (c) is:

(216) ##STR00077##
wherein x is 1;

(217) ##STR00078##
wherein x is 2;

(218) ##STR00079##
wherein x is 3;

(219) ##STR00080##
wherein x is 4;

(220) ##STR00081##
wherein x is 5;
or

(221) ##STR00082##
wherein x is 6;
wherein:

(222) R.sup.c1, R.sup.c2, R.sup.c3, R.sup.c4, R.sup.c5, and R.sup.c6 each independently correspond to the definition and various embodiments of R.sup.c;

(223) R.sup.d1, R.sup.d2, R.sup.d3, R.sup.d4, R.sup.d5, and R.sup.d6 each independently correspond to the definition and various embodiments of R.sup.d;

(224) n1, n2, n3, n4, n5, and n6 each independently correspond to the definition and various embodiments of n;

(225) m1, m2, m3, m4, m5, and m6 each independently correspond to the definition and various embodiments of m;

(226) Y.sup.1, Y.sup.2, Y.sup.3, Y.sup.4, Y.sup.5, and Y.sup.6, each independently correspond to the definition and various embodiments of Y;

(227) and R.sup.z is as defined herein.

(228) In certain embodiments, each of n, n1, n2, n3, n4, n5, and n6 is 0.

(229) In certain embodiments, each of m, m1, m2, m3, m4, m5, and m6 is 0.

(230) In certain embodiments, each of Y, Y.sup.1, Y.sup.2, Y.sup.3, Y.sup.4, Y.sup.5, and Y.sup.6 is O.

(231) In certain embodiments, R.sup.1 is C(O)NH.sub.2; R.sup.2 is hydrogen; R.sup.3 is OH; W is O; R.sup.4 is hydrogen; R.sup.5 is C(O)CH.sub.3; R.sup.6 is OH; R.sup.7 is OH; R.sup.a is hydrogen; and R.sup.b is hydrogen.

(232) The reaction described above takes place in the presence of a galactosyltransferase (GalT) enzyme. In certain embodiments, the GalT is wild type. In certain embodiments, the GalT is a mutant. In certain embodiments, the GalT is GalT Y289L mutant.

(233) Mammalian glycosyltransferases constitute a superfamily of enzymes that are involved in the synthesis of polysaccharides present in glycoproteins and glycolipids. -1,4-Galactosyltransferase 1 (Gal-T1, EC 2.4.1.38) is a bovine enzyme present in milk that uses UDP-galactose (UDP-Gal), and Mn.sup.2+ as cofactor, to transfer galactose to the 4-hydroxyl of N-acetylglucosamine (GlcNAc) to form a -1,4-glycosidic bond (see Ramakrishnan et al. J. Biol. Chem. 2001, 276, 40, 37665 and references cited therein). This enzyme naturally shows some flexibility in the substrate requirements as it transfers glucose, 2-deoxy-glucose, arabinose, and GalNAc from the corresponding UDP-sugars, albeit in low efficiencies. Furthermore, interaction of Gal-T1 with -lactalbumine alters the activity of the enzyme, and glucose can more efficiently be transferred to both glucose and N-acetylglucosamine. In 2002, Qasba and Ramakrishnan reported the structure based design of a mutant of Gal-T1 (i.e. Y289L) that showed both drastically enhanced activity as well as relaxed substrate requirements, with respect to the donor saccharide, while retaining -1,4-selectivity for the acceptor sugar. This protein, a truncated version of the wild type Gal-T1, can be expressed in E. coli and reconstituted in vitro (Ramakrishnan et al. J. Biol. Chem. 2002, 277(23): 20833.). Since this seminal report, multiple applications of this enzyme have been reported in which N-acetyl galactosamine (GalNAc), its 2-carbadeoxy-analog, or N-azidoacetyl galactosamine (GalNAz) were transferred to acceptor substrates. These derivatizations allowed further downstream manipulations that facilitated identification of the acceptor substrate (see, for example, Khidekel et al. J. Am. Chem. Soc. 2003, 125: 16162; Tai et al. J. Am. Chem. Soc. 2004, 126: 10500; Hang et al. Bioorg. Med. Chem. 2005, 13: 5021; Hang et al. Chem. & Biol. 2004, 11: 337; Vocadlo et al. Angew. Chem. Int. Ed. 2004, 43: 5338; Vocadlo et al. Proc. Natl. Acad. Sci. USA 2003, 100: 9116).

(234) In certain embodiments, the compound of Formula (III) is UDP galactose or a derivative thereof. In certain embodiments, the compound of Formula (III) is UDP-Gal. In certain embodiments, the compound of Formula (III) is UDP-GalNAc. In certain embodiments, the compound of Formula (III) is UDP-GalNAz.

(235) In certain embodiments, the enzymatic reaction takes place in a buffer. In certain embodiments, the buffer is TRIS. In certain embodiments, the buffer is HEPES. In certain embodiments, additives are added to the reaction. In certain embodiments, the reaction takes place in the presence of an alkaline phosphatase. In certain embodiments, the reaction takes places in the presence of calf alkaline phosphatase. In certain embodiments, salts are added to the reaction. In certain embodiments, NaCl and/or MgCl.sub.2 are added to the reaction. In certain embodiments, Mn.sup.2+ is added to the reaction. In certain embodiments, MnCl.sub.2 is added to the reaction.

(236) In certain embodiments, the compound of Formula (I) is purified.

(237) In certain embodiments, the compound of Formula (I) is further derivatized. For example, in certain embodiments when R.sup.11 is NHC(O)CH.sub.2N.sub.3, the azide group can be reduced to an amine which can then take part in other reactions, e.g., to form an amide, urea, or thiourea. In other embodiments, the azide group can take part in a cycloaddition, e.g. with an alkyne to form a 1,2,3-triazole. Other reactions include Staudinger ligation, oxime ligation or hydrazone ligation (Dirksen et al., Biocong. Chem. 19:2543-2548 (2008)), inverse electron demand Diels-Alder (e.g., tetrazine ligation (Blackman et al., J. Am. Chem. Soc. 130:13518-13519 (2008))), and [2+2+2]cycloaddition (e.g., quadricyclane ligation (Sletten et al., J. Am. Chem. Soc. 133:17570-17573 (2011))). Methods for such reactions are known in the art. Such further reactions allow for introduction of further diversity or e.g., introduction of a detectable moiety as described in U.S. Provisional Application entitled Methods and Compounds for Identifying Glycosyltransferase Inhibitors, filed on the same day as the present application and incorporated herein by reference.

(238) Additional sugars, such as D ring and/or B ring as described herein, can also be added to Formula (I). Methods for such glycosylation reactions have been described, for example, in PCT Application Publication Nos. WO 2008/021367 and WO 2009/046314, and U.S. Provisional Application entitled Moenomycin A Analogs, Methods of Synthesis, and Uses Thereof, filed on the same day as the present application, each of which is incorporated herein by reference.

(239) In certain embodiments, the G group of a compound of Formula (I) is removed and a different lipid tail is added. See, for example, PCT Application Publication No. WO 2009/046314, and U.S. Provisional Application entitled Moenomycin A Analogs, Methods of Synthesis, and Uses Thereof, filed on the same day as the present application, each of which is incorporated herein by reference.

(240) In certain embodiments, a method of present invention further comprises a deprotection step.

(241) In some embodiments, the compound of Formula (II) is one of the following:

(242) ##STR00083##

(243) In certain embodiments, a compound of formula (I) is:

(244) ##STR00084##

(245) In certain embodiments, the present invention provides a kit comprising a compound of Formula (III) as described herein and a GalT enzyme. In some embodiments, a provided kit comprises a compound of Formula (III) as described herein and a GalT mutant, e.g., GalT Y289L. In certain embodiments, the compound of Formula (III) is UDP galactose or a derivative thereof. In certain embodiments, the compound of Formula (III) is UDP-Gal, UDP-GalNAc, or UDP-GalNAz. In some embodiments, the kit further comprises a buffer. In some embodiments, the kit further comprises an alkaline phosphatase. In certain embodiments, the kit further comprises a salt, e.g., NaCl and/or MgCl.sub.2. In some embodiments, the kit further comprises instructions for use.

(246) These and other aspects of the present invention will be further appreciated upon consideration of the following Examples, which are intended to illustrate certain particular embodiments of the invention but are not intended to limit its scope, as defined by the claims.

EXAMPLES

Chemical Syntheses

(247) All reactions in non-aqueous reaction medium were carried out under an atmosphere of argon, unless otherwise noted. Commercial chemicals were used without prior purification. Solvents were dried by passage over columns filled with activated aluminum oxide (Glass Contour Solvent Systems, SG Water USA, Nashua, N.H., USA).

(248) ##STR00085##

Synthesis of S2 and S3

General procedure for preparation of 2,5-Di-O-alkyl-D-Mannitol

(249) To a stirred suspension of 60% NaH (3 equiv.), washed twice with petroleum ether, in anhydrous DMF (8 mL/mmol-starting material (SM)) was added 1,3:4,6-di-O,O-(4-methoxybenzylidene)-D-mannitol (1 equiv., SM) at room temperature. After being stirred for 30 min, the mixture was treated with a 1.2 M solution of alkylating reagents (2.4 equiv., Br, and methane- or p-toluene-sulfonate for R=allyl, and n-alkyl groups, respectively) in anhydrous DMF and a catalytic amount of tetrabutylammonium iodide for allyl-Br, or 15-Crown-5 for n-alkyl sulfonates. The resulting mixture was stirred for 18 h at rt for allyl-Br and 70 C. for n-alkyl sulfonates, and then poured into sat. aq. NH.sub.4Cl (8 mL/mmol-SM). The immiscible mixture was extracted twice with Et.sub.2O and the combined organic phases were washed with water, brine, dried over MgSO.sub.4, and then concentrated in vacuo. The crude ether was used for the next reaction without further purification.

(250) For allyl derivatives, a stirred solution of the residue in THF-H.sub.2O (4:1, 8 mL/mmol-SM) was treated with AcOH (170 equiv.) at room temperature. After being stirred at 55 C. for 2 d, the mixture was cooled to 0 C. and basified with 4 M aq. K.sub.2CO.sub.3 (90 equiv.). The immiscible mixture was extracted twice with CHCl.sub.3 and the combined organic phases were washed with brine, dried over MgSO.sub.4, and then concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether:EtOAc=1:3 to 0:1) to give 2,5-di-O-allyl-D-mannitol.

(251) For n-alkyl derivatives, a stirred solution of the residue in EtOH (12 mL/mmol-SM) was treated with 3 M aq. HCl (12 equiv.) at room temperature. After being stirred at 70 C. for 3 h, the mixture was cooled to room temperature and basified with 4 M aq. K.sub.2CO.sub.3 (16 equiv.). The immiscible mixture was extracted twice with CHCl.sub.3 and the combined organic phases were washed with brine, dried over MgSO.sub.4, and then concentrated in vacuo. The residue was purified by recrystallization from Et.sub.2O/EtOAc to give 2,5-di-O-n-alkyl-D-mannitol.

Preparation of Methyl 2-O-Alkyl-D-Glycerate

(252) To a 5.5 M solution of 2,5-di-O-alkyl-D-mannitol (1 equiv., SM) in THF-H.sub.2O (9:1) was added NaIO.sub.4 (1.2 equiv.) at room temperature, and the mixture was stirred at 50 C. for 1 h. The resulting inorganic salt was removed by filtration through a pad of silica gel and washed with EtOAc. The filtrate was concentrated in vacuo and the crude aldehyde was used for the next reaction.

(253) To a stirred solution of the residue in t-BuOH (20 mLUmmol-SM) were added 2-methyl-2-butene (100 equiv.) and a solution of 80% NaClO.sub.2 (12 equiv.) and NaH.sub.2PO.sub.4. H.sub.2O (10 equiv.) in H.sub.2O (8 mL/mmol-SM) at 0 C. successively. The resulting yellow mixture was allowed to warm to room temperature for 6 h, during which it turned into clear. Then, the mixture was cooled to 0 C. again and treated with 2.5 M aq. Na.sub.2SO.sub.3 (25 equiv.) to reduce an excess of NaClO.sub.2. The mixture was acidified with 10% aq. citric acid (10 mL/mmol-SM) and extracted twice with CHCl.sub.3 and the combined organic phases were washed with brine, dried over MgSO.sub.4, and then concentrated in vacuo. The crude acid was used for the next reaction without further purification.

(254) To a stirred solution of the residue in anhydrous THF-MeOH (1:1, 10 mL/mmol-SM) was treated with 2 M TMSCHN.sub.2 solution in hexanes (3.2 equiv.) at 0 C. After being stirred for 10 min, the resulting yellow mixture was decolorized by an addition of AcOH (3.2 equiv.) to consume an excess of TMSCHN.sub.2. The mixture was concentrated in vacuo and the residue was purified by silica gel chromatography (petroleum ether:EtOAc=4:1 to 3:2) to give methyl 2-O-alkyl-D-glycerate.

(255) Analytical data for S2: .sup.1H-NMR (500 MHz; CDCl.sub.3): 3.99 (dd, J=6.1, 3.8 Hz, 1H), 3.99 (dd, J=6.1, 3.8 Hz, 1H), 3.79 (d, J=14.1 Hz, 4H), 3.73 (q, J=7.9 Hz, 1H), 3.43 (t, J=11.3 Hz, 1H), 1.65-1.62 (m, 2H), 1.35-1.26 (m, 28H), 0.89 (t, J=7.0 Hz, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3): 171.6, 79.8, 71.7, 63.7, 52.3, 32.2, 29.9 (multiple peaks), 29.8, 29.7, 29.6, 26.2, 22.9, 14.4; HRMS (ESI) calcd for C.sub.20H.sub.40O.sub.4Na.sup.+ [M+Na].sup.+ 367.2819. found 367.2823.

(256) Analytical data for S3: .sup.1H NMR (500 MHz; CDCl.sub.3): 5.66 (dd, J=6.0, 2.1 Hz, 1H), 5.12-5.07 (m, 2H), 4.26-4.22 (m, 1H), 4.10 (d, J=6.5 Hz, 1H), 4.05 (m, 1H), 3.96 (m, 1H), 3.86 (m, 1H), 3.77 (s, 3H), 2.08-1.77 (m, 8H), 1.77 (s, 3H), 1.68 (s, 3H), 1.60 (s, 6H); .sup.13C NMR (125 MHz, CDCl.sub.3): 171.6, 142.5, 136.0, 131.6, 124.5, 123.7, 120.8, 67.2, 63.7, 52.3, 39.9, 32.7, 29.8, 29.7, 26.9, 25.7, 24.1, 23.6, 16.2; HRMS (ESI) calcd for C.sub.19H.sub.32O.sub.4Na.sup.+ [M+Na].sup.+ 347.2193. found 347.2206.

Preparation of 0.2 M Phosphoramidite Solution (S4 and S5)

(257) To a 0.2 M solution of methyl 2-O-alkyl-D-glycerate (1 equiv.) in anhydrous CH.sub.3CN were added N,N-diisopropylethylamine (1.5 equiv.) and ClP(OCE)NiPr.sub.2 (1.2 equiv.) at room temperature successively. The reaction mixture was stirred for 1 h and directly used for the next coupling reaction.

Preparation of S7

(258) To as solution of N-hydroxysuccinimide fluorescein (S6, 15 mg, 21 mol) in DMF (300 L) was added NEt.sub.3 (20 L) and propargyl amine (3.0 mg, 48 mol). After stirring the solution for 24 h the solvent was removed in vacuo and the residue was purified by column chromatography (SiO.sub.2, hexane/EtOAc 2/8) to obtain S7 (11 mg, 27 mol, 84%) as bright orange solid.

(259) .sup.1H NMR (500 MHz, CD.sub.3OD) 8.43 (s, 1H), 8.20 (d, J=9.50 Hz, 1H), 7.31 (d, J=8.0 Hz, 1H), 6.69 (s, 1H), 6.61 (d, J=8.5 Hz, 2H), 6.54 (d, J=9.0 Hz, 2H), 4.59 (s, 1H), 4.21 (d, J=2.0 Hz, 1H), 2.65 (d, J=2.5 Hz, 1H); .sup.13C NMR (500 MHz, CD.sub.3OD) 169.4, 166.8, 152.9, 136.2, 134.4, 129.1, 129.0, 124.7, 123.9, 112.6, 112.2, 109.7, 102.5, 79.3, 78.1, 77.8, 71.2, 29.1.

(260) ##STR00086##

Synthesis of S9

(261) Saccharide S8 (18.7 g, 49.9 mmol, CAS: 176299-96-0) was dissolved in pyridine (160 mL) and cooled to 40 C. Phenylchlorocarbonate (11 mL) was added dropwise to the stirred solution. After 2 h, methanol (11 mL) and toluene (100 mL) were added and the solvent was removed in vacuum. The residue was taken up in EtOAc and washed with HCl (1 M) and NaCl (sat.). The organic layer was dried over MgSO.sub.4 and the solvent was removed in vacuum. Recrystallization from Et.sub.2O yielded the title compound as colorless solid (18.5 g, 75%).

(262) .sup.1H NMR (500 MHz; CDCl.sub.3): 7.54 (d, J=7.6 Hz, 2H), 7.37-3.35 (d, J=1.9 Hz, 5H), 7.22-7.21 (m, 3H), 7.07 (d, J=9.1 Hz, 2H), 6.83 (d, J=9.1 Hz, 2H), 5.55 (s, 1H), 4.91 (dd, J=10.2 Hz, 3.7 Hz, 1H, 4.85 (d, J=7.8 Hz, 1H), 4.53 (d, J=3.3 Hz, 1H), 4.34-4.31 (m, 2H), 4.05 (d, J=11.5 Hz, 1H), 3.76 (s, 3H), 3.51 (s, 1H), 2.89 (d, J=2.8 Hz, 1H); .sup.3C NMR (125 MHz, CDCl.sub.3): 155.9, 153.6, 151.3, 137.8, 129.7, 129.4, 128.4, 126.6, 126.4, 122.0, 121.3, 119.5, 114.8, 102.8, 101.1, 73.2, 69.1, 68.6, 66.5, 55.9; HRMS (ESI) calcd for C.sub.27H.sub.26O.sub.9Na.sup.+ [M+Na].sup.+ 517.1469. found 517.1506.

Synthesis of S10

(263) Sulfoxide S10 was obtained by oxidation of peracyl-N-Troc-phenyl-(S,O)-glucosamine (CAS: 187022-49-7; 9.00 g, 15.1 mmol) with Selectfluor (6.00 g, 16.8 mmol) in MeCN (105 mL) and water (10.5 mL) at room temperature. The reaction was carried out in an open flask. After 1 h the solvent was removed in vacuum, and the residue was taken up in chloroform, washed with NaCl (sat.) and dried over Na.sub.2SO.sub.4. After concentration in vacuo the residue was recrystallized from EtOAc/hexane to yield sulfoxide S10 as an off-white solid (9.10 g, 15.5 mmol, 98%; 1/1 mixture of diastereomers).

Synthesis of S11

(264) In a 100 mL round bottom flask, gylcosyl donor S10 (1.50 g, 2.55 mmol), gylcosyl acceptor S9 (840 mg, 1.70 mmol), 2,6-di-tert-butylpyridine (478 mg, 2.50 mmol), and 4-allyl-1,2-dimethoxybenzene (2.73 g, 15.3 mmol) were combined and dried by azeotropic distillation with benzene. The residue was further dried in vacuum for 30 min before dichloromethane (17 mL) and molecular sieves 4 A (ca. 500 mg) were added. The suspension was stirred at room temperature for 30 min and then cooled to 78 C. Triflic anhydride (285 L, 479 mg, 1.70 mmol) was slowly added and the resulting green solution was stirred for 1.5 h at 78 C. NaHCO.sub.3 (sat., 1 volume) was added, and the mixture was allowed to reach room temperature.

(265) The phases were separated, and the organic phase was washed with NaCl (sat.) and dried over Na.sub.2SO.sub.4. Removal of the solvent in vacuum was followed by column chromatography (SiO.sub.2, toluene/EtOAc 8/2.fwdarw.7/3) to yield the title compound as colorless solid (910 mg, 0.951 mmol, 56%).

(266) .sup.1H NMR (500 MHz; CDCl.sub.3): 7.54 (d, J=7.6 Hz, 2H), 7.39-3.37 (d, J=1.9 Hz, 5H), 7.29-7.26 (m, 3H), 7.02 (d, J=9.1 Hz, 2H), 6.80 (d, J=9.1 Hz, 2H), 5.57 (s, 1H), 5.20-5.00 (m, 4H), 5.85-5.78 (m, 3H), 4.59 (br s, 1H), 4.40-4.22 (m, 3H), 4.21-4.19 (m, 1H), 4.16-4.12 (m, 1H), 3.86-3.75 (m, 2H), 3.77 (s, 3H), 3.59 (br s, 1H), 2.01 (s, 3H), 2.01 (s, 3H), 2.00 (s, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3): 170.99, 170.88, 169.6, 155.63, 154.34, 152.75, 151.5, 151.2, 137.6, 128.9, 129.5, 128.5, 126.6, 121.3, 121.2, 118.8, 118.7, 114.7, 101.9, 101.3, 100.9, 76.2, 74.6, 72.9, 72.4, 72.0, 69.1, 68.4, 66.1, 61.8, 55.9, 20.91, 20.86; HRMS (ESI) calcd for C.sub.42H.sub.44Cl.sub.3NO.sub.18Na.sup.+ [M+Na].sup.+ 978.1516. found 978.1472.

Synthesis of S12

(267) Disaccharide S11 (1.06 g, 1.11 mmol) was dissolved in dichloromethane (22.2 mL) and HSiEt.sub.3 (530 L, 387 mg, 3.33 mmol) and molecular sieves 4 (ca. 500 mg) were added. The suspension was stirred for 30 min at room temperature and then cooled to 78 C. before triflic acid (333 L, 566 mg, 3.77 mmol) was added dropwise. After 2.5 h at 78 C., NaHCO.sub.3 (sat.) was added and the mixture was allowed to reach room temperature. The phases were parted and the aqueous layer was extracted once with dichloromethane. The combined organic layers were washed with brine and dried over Na.sub.2SO.sub.4. Removal of the solvent in vacuum provided the corresponding C6-benzyl ether of S11 in high purity, which was used in the next step without further purification.

(268) The C6-benzyl ether previously obtained (1.15 g, 1.20 mmol) was dissolved in dichloromethane (12 mL) and pyridine (290 L, 284 mg, 3.59 mmol), and the solution was cooled to 40 C. Triflic anhydride (242 L, 406 mg, 1.44 mmol) was slowly added and the mixture was allowed to reach room temperature over 2 h. The reaction mixture was washed with 2 volumes of 0.5 M HCl, water, NaHCO.sub.3 (sat.), and NaCl (sat), and then dried over Na.sub.2SO.sub.4. The solvent was removed in vacuum and the residue was dissolved in toluene (30 mL), and CsOAc (830 mg, 5.48 mmol) and 18-crown-6 (1.21 g, 4.58 mmol) were added. The resulting mixture was vigorously stirred for 14 h and then washed with NaHCO.sub.3 (sat.) and NaCl (sat.). The residue obtained after drying of the solution over Na.sub.2SO.sub.4 and removal of the solvent in vacuum was purified by column chromatography (SiO.sub.2, toluene/EtOAc 85/15) to obtain S12 as colorless solid (668 mg, 667 mol, 55% over 3 steps).

(269) .sup.1H NMR (500 MHz; CDCl.sub.3): 7.41-7.38 (m, 2H), 7.27 (s, 8H), 7.00 (d, J=9.0 Hz, 2H), 6.80 (d, J=8.50 Hz, 2H), 5.31 (t, J=9.6, 1H), 5.20-5.18 (m, 2H), 5.14-5.05 (m, 4H), 5.01 (d, J=8.1 Hz, 1H), 4.68 (d, J=12.0 Hz, 1H), 4.56 (d, J=11.8, 1H), 4.47 (d, J=11.9, 1H), 4.27 (d, J=12.2 Hz, 1H), 4.18 (dd, J=11.9, 3.4 Hz, 1H), 4.07 (td, J=8.2, 1.4 Hz, 1H), 3.77 (s, 3H), 3.67-3.60 (m, 3H), 3.59-3.57 (m, 1H), 2.01 (s, 3H), 2.01 (s, 3H), 2.00 (s, 3H), 1.95 (s, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3): 170.93, 170.86, 170.11, 169.7, 155.69, 154.21, 153.32, 151.23, 151.12, 138.10, 137.91, 129.84, 129.27, 128.61, 128.46, 128.03, 127.98, 126.55, 125.52, 121.27, 118.12, 114.82, 114.79, 101.14, 100.44, 95.67, 79.60, 78.88, 74.53, 73.81, 73.22, 72.13, 72.00, 69.56, 68.81, 68.46, 61.72, 56.72, 55.91, 21.70, 20.91, 20.85, 20.81; HRMS (ESI) calcd for C.sub.44H.sub.48Cl.sub.3NO.sub.19Na.sup.+ [M+Na].sup.+ 1022.1779. found 1022.1766.

Synthesis of S13

(270) In a 100 ml round bottom flask, disaccharide S12 (283 mg, 283 mol) was dissolved in methanol (20 mL), and 10% Pd/C (100 mg) was added. The atmosphere above the solution was exchanged to H.sub.2, and the solution was stirred vigorously. After 45 min the suspension was filtered through Celite and the filtrate was concentrated in vacuo. The residue was dissolved in dichloromethane (3.4 mL) and water (1.7 mL), and PhI(OAc).sub.2 (237 mg, 756 mol) and TEMPO (9.0 mg, 57 mol) were added. After stirring the mixture for 2 h, the reaction was quenched by addition of Na.sub.2S.sub.2O.sub.3 (sat.) and the solution was partitioned between dichloromethane and water. The organic layer was washed with NaCl (sat.) and dried over Na.sub.2SO.sub.4. The solvent was removed in vacuo, and the residue was taken up in THF (5.7 mL). This solution was cooled to 40 C., N-methylmorpholine (78 L, 72 mg, 0.71 mmol) was added and the mixture was treated with isobutyl chloroformate (93 L, 97 mg, 0.71 mmol). After 5 min, NH.sub.3 (2.0 M in .sup.iPrOH) was added and the mixture was stirred at room temperature for 24 h. Removal of the solvent in vacuum and column chromatography (SiO.sub.2, CHCl.sub.3/EtOH 99/1.fwdarw.95/5.fwdarw.9/1) yielded the title compound as colorless flakes (111 mg, 131 mol, 46% over 3 steps).

(271) .sup.1H NMR (500 MHz; CDCl.sub.3/CD.sub.3OD 9/1): 6.92 (d, J=8.9 Hz, 2H), 6.76 (d, J=9.0 Hz, 2H), 5.22 (t, J=9.9 Hz, 1H), 5.16-5.12 (m, 2H), 4.98 (dd, J=10.0 Hz, 2H), 4.90 (d, J=8.4 Hz, 1H), 4.87 (d, J=12.1 Hz, 1H), 4.49 (d, J=12.1 Hz, 1H), 4.09 (dd, J=12.3, 3.7 Hz, 1H), 3.99 (d, J=8.9 Hz, 1H), 3.88 (t, J=7.1 Hz, 1H), 3.78 (d, J=11.6 Hz, 1H), 3.71 (s, 3H), 3.62 (d, J=9.9 Hz, 1H), 3.55-3.48 (m, 5H), 2.00 (s, 3H), 1.94 (s, 3H), 1.93 (s, 3H), 1.92 (s, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3/CD.sub.3OD 9/1): 171.20, 170.92, 170.6, 170.3, 170.0, 156.4, 155.8, 154.8, 150.7, 118.3, 114.9, 101.1, 100.1, 95.8, 79.4, 74.5, 73.9, 72.4, 71.8, 69.6, 68.7, 61.9, 56.5, 55.8, 49.7, 49.5, 49.4, 49.2, 48.9, 48.7, 20.7, 20.7; HRMS (ESI) calcd for C.sub.31H.sub.38Cl.sub.3N.sub.3O.sub.18Na.sup.+ [M+Na].sup.+ 868.1109. found 868.1067.

Synthesis of S14

(272) Disaccharide S13 (167 mg, 187 mol) was dissolved in MeCN (8 mL) and water (2 mL), and cerium(IV) ammonium nitrate (542 mg, 989 mol) was added. The mixture was stirred at room temperature for 1.5 h and then concentrated in vacuo. Purification of the residue by column chromatography (SiO.sub.2, CHCl.sub.3/EtOH 9/1.fwdarw.4/1) gave the lactol of S13 as colorless solid (115 mg, 79%). This lactol (42.9 mg, 57.9 mol) was further dried by azeotropic distillation with toluene (2), dissolved in tetrazole solution (0.34 M in MeCN, 1.0 mL), and stirred with molecular sieves 4 for 30 min at room temperature and 30 min at 0 C. A solution of S4 (0.2 M in MeCN, 0.58 mL) was added and the mixture was stirred at 0 C. for 2 h before .sup.tBuOOH (5.5 M in decane, 127 L, 699 mol) was added. After 1 h at 0 C. P(OMe).sub.3 (82 L, 86 mg, 695 mol) was added and the suspension was filtered through a pad of Celite. Evaporation of the solvent in vacuo and column chromatographic purification (C18, gradient 30-100% MeCN/H.sub.2O) of the residue gave phosphoglycerate S14 (32.5 mg, 27.1 mol, 47%) as colorless solid as a 1/1 mixture of phosphate diastereomers.

(273) Analytical data for one diastereomer: .sup.1H NMR (500 MHz; CD.sub.3OD): 6.03 (dd, J=6.3, 3.2 Hz, 1H), 5.27-5.20 (m, 3H), 5.07 (t, J=9.7 Hz, 1H), 5.02 (d, J=12.3 Hz, 1H), 4.78 (d, J=8.5 Hz, 1H), 4.61 (d, J=12.3 Hz, 1H), 4.46-4.42 (m, 4H), 4.45-4.41 (m, 4H), 4.22 (dd, J=12.3, 2.2 Hz, 1H), 4.00 (dd, J=6.9, 3.3 Hz, 1H), 3.87-3.83 (m, 2H), 3.82 (s, 3H), 3.72-3.65 (m, 2H), 3.59-3.55 (m, 1H), 3.01 (t, J=6.0 Hz, 2H), 2.10 (s, 3H), 2.04 (s, 3H), 2.01 (s, 3H), 1.96 (s, 3H), 1.66 (t, J=7.2 Hz, 2H), 1.30 (m, 28H), 0.91 (t, J=7.0 Hz, 3H); HSQC (.sup.13C signals, 125 MHz, CD.sub.3OD): 102.7, 97.2; 77.9, 77.6, 74.2, 74.1, 72.2, 72.0, 71.5, 71.4, 70.4, 70.3, 69.5, 68.8, 68.2, 68.1, 63.6, 61.9, 61.8, 55.6, 51.8, 48.1, 31.6, 29.6 (multiple peaks), 25.8, 22.5, 19.6, 19.3, 19.3, 19.3, 18.9, 13.2; .sup.31P (162 MHz; CD.sub.3OD): 3.16; HRMS (ESI) calcd for C.sub.47H.sub.74Cl.sub.3N.sub.4O.sub.23PNa.sup.+ [M+Na].sup.+ 1223.3410. found 1223.3337.

Synthesis of S15

(274) To a solution of phosphoglycerate S14 (113 mg, 94.2 mol) in THF (3 mL), Ac.sub.2O (1 mL), and AcOH (2 mL) was added activated Zn (653 mg, 9.99 mmol), and the solution was stirred at room temperature for 10 h. The slurry was filtered through a pad of SiO.sub.2 and the residue was thoroughly washed with CHCl.sub.3/EtOH 2/1 (100 mL). The filtrate was concentrated, and the residue was dissolved in THF (19 mL), H.sub.2O.sub.2 (30%, 4.8 mL) and aqueous LiOH solution (1 M, 1.9 mL). After stirring the solution at 0 C. for 2 h, DOWEX50WX2-100 resin (HPy+ form, 0.8 g) was added, and the mixture was stirred for 2 h. The resin was filtered off by passing the reaction mixture through a cotton plug. Chromatographic purification (C18, gradient 20-50% MeCN in 0.2% NH.sub.4HCO.sub.3 aq.) of the residue obtained after concentration of the filtrate yielded disaccharide S15 (50.7 mg, 61.0 mol, 63%).

(275) HPLC/MS retention time: 11.9 min (Phenomenex Luna, 3u-C18 502 mm.sup.2 3 micron, 0.3 ml/min, gradient 30-75% MeCN+0.1% HCO.sub.2H in H.sub.2O+0.1% HCO.sub.2H over 6 min, then to 99% MeCN+0.1% HCO.sub.2H over 5 min) LRMS (ESI) calcd for C.sub.34H.sub.61N.sub.3O.sub.18P.sup. [MH].sup. 830.4. found 830.3; .sup.1H NMR (500 MHz; CD.sub.3OD): 5.98 (dd, J=7.1, 3.1 Hz, 1H), 5.05 (t, J=9.7 Hz, 1H), 4.58 (d, J=8.4 Hz, 1H), 4.36 (d, J=10.0 Hz, 1H), 4.26-4.22 (m, 1H), 4.18-4.08 (m, 1H), 4.00-3.96 (m, 1H), 3.87-3.84 (m, 1H), 3.76-3.71 (m, 3H), 3.71 (s, 3H), 3.68-3.64 (m, 3H), 3.53-3.47 (m, 3H), 3.36-3.29 (m, 4H), 2.04 (s, 3H), 1.66-1.64 (m, 2H), 1.30 (m, 28H), 0.91 (t, J=7.0 Hz, 3H); .sup.13C NMR (125 MHz, CD.sub.3OD): ; 175.6, 173.0, 158.2, 103.0, 95.5, 79.6, 76.9, 74.5, 73.2, 71.4, 70.8, 70.6, 70.5, 70.2, 66.9, 66.8, 61.6, 61.1, 59.9, 56.1, 31.9, 29.6, 29.6, 29.5, 29.3, 26.0, 22.6, 22.2, 13.3; .sup.31P (162 MHz; D.sub.6-DMSO): 2.31; HRMS (ESI) calcd for C.sub.34H.sub.63N.sub.3O.sub.18P.sup.+[M+H].sup.+ 832.3839. found 832.7735.

Synthesis of S16 (CMG121)

(276) To a solution of TRIS (50 mM in H.sub.2O, pH=8.0, 2.5 mL) were sequentially added MnCl.sub.2 (50 mM in H.sub.2O, 2.5 mL), H.sub.2O (5 mL), GalT Y289L (Ramakrishanan et al. J. Biol Chem. 2002, 277, 20833; 1 mg/mL in 50 mM TRIS buffer, pH=8.0, 1.88 mL,), UDP-N-azidoacetylgalactosamine (UDP-GalNAz, prepared according to Hang et al. J. Am. Chem. Soc. 2003, 126, 6; 20 mM in H.sub.2O, 500 L), disaccharide S15 (10 mM in H.sub.2O, 125 L), and calf intestinal alkaline phosphatase (CIP, 1000 U, 25 L, Roche Diagnostics GmbH, Mannheim, Germany). The mixture was gently mixed and kept at 37 C. for 60 h. MeOH (7.5 mL) was added, and the mixture was vortexed and centrifuged (15 min at 5000g) to pellet precipitated proteins. The supernatant was concentrated in vacuo, and the residue obtained was loaded onto a Phenomenex Strata C-18 column preequilibrated with H.sub.2O. The column was eluted with H.sub.2O to obtain unreacted UDP-GalNAz. Elusion with H.sub.2O/MeOH 1/9 provided the desired trisaccharide in near quantitative yield.

(277) HPLC retention time: 11.0 min (Phenomenex Luna, 3u-C18 502 mm.sup.2 3 micron, 0.3 mL/min, gradient 30-75% MeCN+0.1% HCO.sub.2H in H.sub.2O+0.1% HCO.sub.2H over 6 min, then to 99% MeCN+0.1% HCO.sub.2H over 5 min) LRMS (ESI) calcd for C.sub.42H.sub.73N.sub.7O.sub.23P.sup. [MH].sup. 1074.5. found 1074.3.

(278) The GalNAz-trisaccharide previously obtained (4.0 mg, 3.8 mol) was dissolved in DMF (400 L) and CuSO.sub.4 (0.9 M in H.sub.2O, 4.0 L, 3.6 mol) and Na-ascorbate (1.8 M in H.sub.2O, 4.0 L, 7.2 mol) were added. The mixture was stirred at room temperature, and after 24 h another portion of CuSO.sub.4 (0.9 M in H.sub.2O, 4.0 L, 3.6 mol) and Na-ascorbate (1.8 M in H.sub.2O, 4.0 L, 7.2 mol) was added. After 48 h the solution was concentrated in vacuo and the residue was purified by column chromatography (C-18, gradient 30-90% MeOH in H.sub.2O; then 10% 2 M NH.sub.3 in MeOH to elute the product) to obtain the title compound (4.9 mg, 3.3 mol, 87%).

(279) HPLC/MS retention time: 10.8 min (Phenomenex Luna, 3u-C18 502 mm.sup.2 3 micron, 0.3 mL/min, gradient 30-75% MeCN+0.1% HCO.sub.2H in H.sub.2O+0.1% HCO.sub.2H over 6 min, then to 99% MeCN+0.1% HCO.sub.2H over 5 min); LRMS (ESI) calcd for C.sub.66H.sub.88N.sub.8O.sub.29P.sup. [MH].sup. 1487.5. found 1487.3; .sup.1H NMR (600 MHz; CD.sub.3OD): 8.51 (s, 1H), 8.28-8.27 (m, 1H), 8.11 (s, 1H), 7.35 (d, J=7.5 Hz, 1H), 6.72 (m, 2H), 6.65 (d, J=8.5 Hz, 2H), 6.60-6.59 (m, 2H), 6.01 (br s, 1H), 5.29 (br s, 1H), 5.05 (t, J=6.8 Hz, 1H), 4.77 (m, 2H), 4.61-4.56 (m, 2H), 4.38 (d, J=7.5 Hz, 1H), 4.29-4.25 (m, 2H), 4.15-4.05 (m, 2H), 4.05-3.99 (m, 2H), 3.86-3.76 (m, 4H), 3.76-3.60 (m, 13H), 3.48 (br s, 1H), 2.02 (s, 3H), 1.65-1.64 (m, 6H), 1.48-1.42 (m, 12H), 1.31 (m, 38H), 0.92 (t, J=5.4 Hz, 3H); HSQC (.sup.13C signals, 125 MHz, CD.sub.3OD): 129.1, 124.1, 112.9, 102.4, 102.3, 79.5, 75.8, 75.0, 73.3, 73.0, 71.5, 71.2, 70.5, 70.0, 68.3, 67.0, 61.2, 53.4, 52.0, 35.1, 31.8, 29.5, 26.0, 23.5, 22.6, 21.9, 13.1; HRMS (ESI) calcd for C.sub.66H.sub.90N.sub.8O.sub.29P.sup.+ [M+H].sup.+ 1489.5546. found 1489.5420.

(280) ##STR00087##

Synthesis of S17

(281) Disaccharide S11 (650 mg, 679 mol) was dissolved in dichloromethane (13.6 mL) and HSiEt.sub.3 (325 L, 237 mg, 2.04 mmol) and molecular sieves 3 (ca. 250 mg) were added. The suspension was stirred for 30 min at room temperature and then cooled to 78 C. before triflic acid (204 L, 347 mg, 2.31 mmol) was added dropwise. After 2.5 h at 78 C., NaHCO.sub.3 (sat.) was added and the mixture was allowed to reach room temperature. The phases were parted and the aqueous layer was extracted once with dichloromethane. The combined organic layers were washed with brine and dried over Na.sub.2SO.sub.4. Removal of the solvent in vacuo provided the corresponding C6-benzyl ether of S11 in high purity, which was dissolved in dichloromethane (3.4 mL), and pyridine (164 L, 161 mg, 2.04 mmol), DMAP (8.3 mg, 0.07 mmol), and Ac.sub.2O (94 L, 104 mg, 1.02 mmol) were added. After 3 h at room temperature the reaction was diluted with dichloromethane and washed with HCl (1 M), H.sub.2O, NaHCO.sub.3 (sat.), and NaCl (sat.). The organic layers were dried over Na.sub.2SO.sub.4 and then concentrated in vacuo. Purification of the residue yielded acetate S17 (539 mg, 538 mol, 79% over 2 steps) as colorless solid.

(282) .sup.1H NMR (500 MHz; CDCl.sub.3): 7.39 (t, J=7.8 Hz, 2H), 7.34-7.25 (m, 8H), 7.01 (d, J=9.0 Hz, 2H), 6.79 (d, J=9.0 Hz, 2H), 5.64 (d, J=2.5 Hz, 1H), 5.26 (t, J=10.1 Hz, 1H), 5.14-5.10 (m, 2H), 5.01 (d, J=7.2 Hz, 1H), 4.90-4.88 (m, 2H), 4.81 (d, J=12.2 Hz, 1H), 4.56 (d, J=12.0 Hz, 1H), 4.44 (d, J=12.0 Hz, 1H), 4.29 (d, J=12.2 Hz, 1H), 4.23 (dd, J=12.2, 3.3 Hz, 1H), 4.11 (dd, J=9.8, 7.7 Hz, 1H), 3.94 (t, J=6.4 Hz, 1H), 3.90-3.86 (m, 2H), 3.77 (s, 3H), 3.70-3.66 (m, 2H), 3.59-3.55 (m, 2H), 2.11 (s, 3H), 2.03 (s, 3H), 2.02 (s, 3H), 2.01 (s, 3H); .sup.13C NMR (125 MHz, CDCl.sub.3): 171.1, 170.9, 170.5, 169.6, 155.7, 154.2, 152.7, 151.5, 151.2, 138.1, 137.7, 129.8, 129.3, 128.7, 128.5, 128.2, 128.1, 126.6, 125.6, 121.5, 118.3, 114.8, 101.9, 100.8, 95.7, 74.5, 73.8, 72.1, 72.0, 68.4, 67.7, 67.2, 61.8, 56.9, 55.9, 21.0, 20.9, 20.8, 20.8; HRMS (ESI) calcd for C.sub.44H.sub.48Cl.sub.3NO.sub.19Na.sup.+ [M+Na].sup.+ 1022.1779. found 1022.1707.

Synthesis of S18

(283) In a 10 mL round bottom flask S17 (56.4 mg, 56.3 mol) was dissolved in a solution of 1% trichloroacetic acid in methanol (2.8 mL, 2.4 equiv. of TCA), 10% Pd/C (11.9 mg) was added and the atmosphere above the solution was exchanged to H.sub.2. After stirring for 15 min the solution was filtered through a pad of Celite and poured into NaHCO.sub.3 (sat.). The mixture was extracted with EtOAc (2), washed with NaCl (sat.), and dried over MgSO.sub.4. Evaporation of the solvent in vacuum yielded the free C6-alcohol in quantitative yield.

(284) The alcohol previously obtained (51.3 mg, 56.3 mol) was dissolved in dichloromethane (0.2 mL) and water (0.1 mL). After addition of TEMPO (1.8 mg, 1.2 mol) and diacetoxy iodobenzene (45.0 mg, 140 mol) the mixture was stirred at room temperature for 1.5 h. Na.sub.2S.sub.2O.sub.3 (sat.) solution was added, and the reaction mixture was extracted with EtOAc (2). The combined organic layers were washed with NaCl (sat.) and dried over MgSO.sub.4. The residue obtained after concentration of the solution in vacuum was purified by column chromatography (SiO.sub.2, petroleum ether/EtOAc/1% AcOH 2/1.fwdarw.1/4) to obtain pure C6-carboxylic acid (36.8 mg, 39.8 mol, 71% over 2 steps).

(285) The C6-carboxylic acid (22.0 mg, 23.8 mol) was dissolved in THF (0.6 mL) and N-methyl-morpholine (5.2 L, 47 mol), and the solution was cooled to 30 C. before isobutylchloroformate (6.2 L, 47 mol) was added. After 30 min the turbid mixture was treated with 7 M NH.sub.3 solution in MeOH (14 L) and stirred at 0 C. for another 30 min. The mixture was poured into NH.sub.4Cl and extracted with EtOAc (2). The organic layers were washed with NaCl (sat.) and dried over MgSO.sub.4 before they were concentrated in vacuo. Column chromatographic purification of the residue (SiO.sub.2, petroleum ether/EtOAc 2/1.fwdarw.1/4) gave S18 as colorless solid (21.8 mg, 23.6 mol, 99%).

Synthesis of S19

(286) Deprotection of the PMP group was achieved by treatment of a solution of S18 (21.8 mg, 23.8 mol) in MeCN (1.2 mL) and water (0.3 mL) with cerium (IV) ammonium nitrate (40.2 mg, 73.3 mol). After stirring at room temperature for 1 h the mixture was concentrated in vacuo, and the residue was purified by column chromatography (SiO.sub.2, CHCl.sub.3/MeOH 98/2.fwdarw.95/5) to give the free lactol as colorless solid (12.8 mg, 15.6 mol, 66%). This material was dried by azeotropic distillation with toluene (2), dissolved in a solution of tetrazole in MeCN (0.34 M, 0.28 mL), and molecular sieves 3 (43 mg) were added. The mixture was stirred at room temperature for 15 min and then cooled to 0 C. before a solution of S5 was added (0.2 M in MeCN, 0.15 mL). After 1 h at 0 C., .sup.tBuOOH (5.5 M in decane, 23 L) was added, and the suspension was stirred for another hour before P(OMe).sub.3 (22 L, 186 mol) was added. The mixture was filtered over Celite, and the residue was concentrated in vacuo. Purification of the residue by column chromatography (SiO.sub.2, CHCl.sub.3/MeOH 97/3.fwdarw.96/4) gave S19 as colorless solid as a mixture of phosphate diastereomers (16.0 mg, 12.7 mol, 81%).

Synthesis of S20

(287) Phosphoglycerate S19 (7.6 mg, 6.0 mol) was dissolved in a mixture of THF (0.3 mL), Ac.sub.2O (0.1 mL), and AcOH (0.2 mL) and activated zinc (70.1 mg) was added in portions over the course of 1.5 d. The suspension was filtered through a pad of Celite, concentrated in vacuo, and subjected to column chromatography (SiO.sub.2, CHCl.sub.3/MeOH 96/4). The product obtained was dissolved in a mixture of THF (0.66 mL), MeOH (0.22 mL), and H.sub.2O (0.22 mL) and LiOH (1 M in H.sub.2O, 66 L, 66 mol) was added. After stirring at room temperature for 1.5 h, AcOH (4 L, 7 mol) was added, and the solution was concentrated in vacuo. Purification of the residue by column chromatography (C18, gradient 25-75% MeOH in H.sub.2O+0.1% AcOH) gave S20 (3.3 mg, 4.3 mol, 71% over 2 steps) as colorless solid.

Synthesis of S21

(288) To a solution (2 mL) containing HEPES (50 mM, pH=7.5), NaCl (100 mM), MnCl.sub.2 (1 mM), S20 (1 mM), UDP-N-azidoacetylgalactosamine (UDP-GalNAz, prepared according to Hang et al. J. Am. Chem. Soc. 2003, 126, 6; 2 mM) were added calf intestinal alkaline phosphatase (1000 U, 5 L, Roche Diagnostics GmbH, Mannheim, Germany) and GalT Y289L (Ramakrishanan et al. J. Biol Chem. 2002, 277, 20833, 150 g), and the mixture was incubated at 37 C. for 2 h. The reaction was quenched by precipitation of the proteins by addition of MeOH (4 mL), was centrifuged, and the supernatant was passed over a 30 mg Strata-X C18 column (Phenomenex). The column was eluted with water (2 mL) to rinse off salts, UMP, and UDP-GalNAz, and the desired trisaccharide S21 was eluted with MeOH/H.sub.2O 8/2. The material obtained was directly used in the next reaction.

Synthesis of S22

(289) Azide S21 (3.8 mg, 3.8 mol) was dissolved in a mixture of MeOH (210 L), water (10 L), and HOAc (2 M, 2 L) and Pd(OH).sub.2/C (1 mg) was added. The atmosphere above the solution was exchanged for H.sub.2, and the mixture was stirred vigorously for 48 h. The mixture was filtered over a plug of Celite, the residue was thoroughly washed with MeOH/H.sub.2O 4/1, and the filtrate was concentrated in vacuo. The residue obtained was suspended in DMF (76 L) and treated with diisopropylethylamine (5.3 L) and fluorescein isothiocyanate (1.7 mg, 4.6 mol). After 16 h at room temperature, the solution was concentrated in vacuum, and the residue was purified by column chromatography (SiO.sub.2, CHCl.sub.3/MeOH+0.1% AcOH 9/1.fwdarw.8/2; then CHCl.sub.3/MeOH/H.sub.2O 60/40/8) to obtain the title compound (2.0 mg, 1.5 mol, 40% over 2 steps) as orange solid.

(290) HPLC/MS retention time: 11.6 min (Phenomenex Luna, 3u-C18 502 mm.sup.2 3 micron, 0.3 mL/min, gradient 25-45% MeCN+0.1% HCO.sub.2H in H.sub.2O+0.1% HCO.sub.2H over 6 min, then to 99% MeCN+0.1% HCO.sub.2H over 6.5 min); LRMS (ESI) calcd for C.sub.61H.sub.77N.sub.5O.sub.27PS.sup. [MH].sup. 1374.4. found 1374.3; HRMS (ESI) calcd for C.sub.61H.sub.79N.sub.5O.sub.27PS.sup.+ [M+H].sup.+ 1376.4416. found 1376.4224.

(291) ##STR00088##

(292) All small scale (20 uL) reactions of GalT(Y289L) were run in 50 mM HEPES pH=7.5, 100 mM NaCl, 1 mM MnCl.sub.2 with 0.2 L calf alkaline phosphatase, 40 M disaccharide, and 100 M UDP-GalNAz. The reaction was initiated by the addition of 1.5 g of GalT(Y289L), incubated at 37 C. for 2 h, and quenched with 2 volumes of MeOH. After centrifuging at 10 k rpm for 2 minutes, the reaction was analyzed by LC-MS QTOF.

(293) For scale up of the GalT(Y289L) reactions, 2 mL reactions were set up in 50 mM HEPES pH=7.5, 100 mM NaCl, 1 mM MnCl.sub.2 with 5 L of calf alkaline phosphatase, 1 mM disaccharide, and 2 mM UDP-GalNAz. These reactions were initiated by the addition of 0.15 mg of GalT(Y289L), incubated at 37 C. for 2 h, and stored at 20 C. until purification. The resultant trisaccharide was purified by revered phase chromatography. Each 2 mL reaction was centrifuged and loaded onto a 30 mg Strata-X C18 column (Phenomenex). The protein pellet was washed with 2 mL water, which was then also run on the column. The column was further washed with another 2 ml of water, then, the trisaccharide was eluted in 80% methanol. Reactions were analyzed by LC-MS QTOF:

(294) TABLE-US-00001 substrate (M H) product (M H) % Compound expected observed ppm expected observed ppm conversion 14 830.3693 830.3670 2.79 1074.4500 1074.4457 3.07 85% 15 829.3853 829.3844 1.09 1073.4660 1073.4648 1.12 67% 16 828.4012 828.4005 0.85 1072.4820 1072.4825 0.47 52% 17 865.3740 865.3707 3.81 1109.4548 1109.4515 3.05 88% 18 797.3115 797.3071 5.48 1041.3922 1041.3919 0.37 100%

OTHER EMBODIMENTS

(295) This application refers to various issued patents, published patent applications, journal articles, books, manuals, and other publications, all of which are incorporated herein by reference.

(296) The foregoing written specification is considered to be sufficient to enable one skilled in the art to practice the invention. The present invention is not to be limited in scope by examples provided, since the examples are intended as a single illustration of one aspect of the invention and other functionally equivalent embodiments are within the scope of the invention. Various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims. The advantages and objects of the invention are not necessarily encompassed by each embodiment of the invention.