ORALLY AVAILABLE COMPOUNDS, A PROCESS FOR PREPARING THE SAME AND THEIR USES AS ANTI-ADHESIVE DRUGS FOR TREATING E. COLI INDUCED INFLAMMATORY BOWEL DISEASES SUCH AS CROHN'S DISEASE

20190374559 · 2019-12-12

Inventors

Cpc classification

International classification

Abstract

Orally available compounds, a process for preparing the same and their uses as anti-adhesive drugs for treating E. coli induced inflammatory bowel diseases such as crohn's disease.

Claims

1. Method of treatment or prevention of inflammatory bowel disease, or Crohn disease or ulcerative colitis, comprising administering to a subject in need thereof an effective amount of a compound of the following formula (I): ##STR00273## wherein: X represents NH, O, S or CH.sub.2; n represents an integer comprised from 3 to 7, or n being equal to 5; Y represents a group selected from: ##STR00274## Z representing O, S or NH; R representing: H, a linear or branched (C.sub.1-C.sub.7)-alkyl, or methyl, ethyl, isopropyl or isobutyl, a group of formula (CH.sub.2).sub.iX(CH.sub.2).sub.jH, wherein X represents O, S or NH, i is an integer from 1 to 7, and j is an integer from 0 to 7, or a group CH.sub.2OCH.sub.3, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7) -cycloalkyl, a (C.sub.5-C.sub.7) -cycloalkenyl, a (C.sub.3-C.sub.7) -heterocycloalkyl, a (C.sub.5-C.sub.7) -heterocycloalkenyl, an aryl, said aryl being an aromatic or heteroaromatic group, an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, CF.sub.3, adamantyl, CHRa-NH.sub.2, wherein Ra represents the side chain of a proteinogenic aminoacid, a cyclodextrin, or a cyclodextrin chosen from -cyclodextrin (-CD), -cyclodextrin (-CD), -cyclodextrin (-CD) and their derivatives, or alkylated -cyclodextrins, alkylated -cyclodextrins and alkylated -cyclodextrins, or a cyclodextrin of one of the following formulae: ##STR00275## said (C.sub.1-C.sub.7) -alkyl, group of formula (CH.sub.2).sub.iX(CH.sub.2).sub.jH, (C.sub.2-C.sub.7) -alkenyl, (C.sub.2-C.sub.7) -alkynyl, (C.sub.3-C.sub.7) -cycloalkyl, (C.sub.5-C.sub.7)-cycloalkenyl, (C.sub.3-C.sub.7)-heterocycloalkyl, (C.sub.5-C.sub.7)-heterocycloalkenyl, CO-(C.sub.1-C.sub.7)-alkyl, CO.sub.2-(C.sub.1-C.sub.7)-alkyl, CONH-(C.sub.1-C.sub.7)-alkyl, aryl, alkyl aryl, CO-aryl and cyclodextrin being substituted or not by one or more substituent(s), each independently selected from: a linear or branched (C.sub.1-C.sub.7)-alkyl, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7)-cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, wherein the aryl is an aromatic or heteroaromatic group an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CHO, a CO-(C.sub.1-C.sub.7) -alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, a halogen selected from the group comprising F, Cl, Br, and I, CF.sub.3 OR.sub.a, wherein R.sub.a represents: H, a linear or branched (C.sub.1-C.sub.7) -alkyl, a (C.sub.3-C.sub.7)-cycloalkyl, CO-(C.sub.1-C.sub.7)-alkyl, or CO-aryl, wherein aryl is an aromatic or heteroaromatic group, NR.sub.bR.sub.c, wherein R.sub.b and R.sub.c represent independently from each other: H, a linear or branched (C.sub.1-C.sub.7) -alkyl, a (C.sub.3-C.sub.7)-cycloalkyl, CO-(C.sub.1-C.sub.7)-alkyl, or CO-aryl, wherein aryl is an aromatic or heteroaromatic group, NO.sub.2, CN, SO.sub.3H or one of its salts, or SO.sub.3Na; and its pharmaceutically acceptable salts, provided that when R represents CHRa-NH.sub.2, then Y can only represent the following group (a): ##STR00276##

2. Method according to claim 1, comprising administering to a subject in need thereof an effective amount of a compound formula (I), wherein R is R.sub.1, R.sub.1 representing: H a linear or branched (C.sub.1-C.sub.7)-alkyl, or isopropyl, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7) -cycloalkyl, a (C.sub.5-C.sub.7) -cycloalkenyl, a (C.sub.3-C.sub.7) -heterocycloalkyl, a (C.sub.5-C.sub.7) -heterocycloalkenyl, an aryl, said aryl being an aromatic or heteroaromatic group, an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CO-(C.sub.1-C.sub.7) -alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7) -alkyl, a CONH-(C.sub.1-C.sub.7) -alkyl, CF.sub.3, Adamantyl, CHRa-NH.sub.2, wherein Ra represents the side chain of a proteinogenic aminoacid.

3. Method according to claim 1, comprising administering to a subject in need thereof an effective amount of a compound formula (I), wherein Y represents: ##STR00277## of following formula (I-1a): ##STR00278## X and n being as previously defined, R.sub.1 representing: H a linear or branched (C.sub.1-C.sub.7)-alkyl, or isopropyl, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7)-cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, said aryl being an aromatic or heteroaromatic group, an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, CF.sub.3, Adamantyl, CHRa-NH.sub.2, wherein Ra represents the side chain of a proteinogenic aminoacid.

4. Method according to claim 1, comprising administering to a subject in need thereof an effective amount of a compound formula (I), wherein Y represents: ##STR00279## of following formula (I-1b): ##STR00280## X and n being as previously defined, R.sub.1 representing: H a linear or branched (C.sub.1-C.sub.7)-alkyl, or isopropyl, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7) -cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, said aryl being an aromatic or heteroaromatic group, an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, CF.sub.3, Adamantyl, CHRa-NH.sub.2, wherein Ra represents the side chain of a proteinogenic aminoacid.

5. Method according to claim 1, comprising administering to a subject in need thereof an effective amount of a compound formula (I), wherein Y represents: ##STR00281## of following formula (I-1c): ##STR00282## X, Z and n being as previously defined, R.sub.1 representing: H a linear or branched (C.sub.1-C.sub.7)-alkyl, or isopropyl, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7)-cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, said aryl being an aromatic or heteroaromatic group, an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, CF.sub.3 Adamantyl, CHRa-NH.sub.2, wherein Ra represents the side chain of a proteinogenic aminoacid.

6. Method according to claim 1, comprising administering to a subject in need thereof an effective amount of a compound formula (I), wherein R is R.sub.2, R.sub.2 representing a cyclodextrin, or a cyclodextrin chosen from -cyclodextrin (-CD), (-cyclodextrin (-CD), -cyclodextrin (-CD) and their derivatives, or alkylated -cyclodextrins, alkylated -cyclodextrins and alkylated -cyclodextrins, or a -cyclodextrin of the following formula: ##STR00283##

7. Method according to claim 1, comprising administering to a subject in need thereof an effective amount of a compound formula (I), wherein Y represents: ##STR00284## of following formula (I-2c): ##STR00285## X, n, Z and R.sub.2 representing a cyclodextrin, or a cyclodextrin chosen from -cyclodextrin (-CD), -cyclodextrin (-CD), -cyclodextrin (-CD) and their derivatives, or alkylated -cyclodextrins, alkylated -cyclodextrins and alkylated -cyclodextrins, or a -cyclodextrin of the following formula: ##STR00286##

8. Method according to claim 1, comprising administering to a subject in need thereof an effective amount of a compound selected from the group consisting of: ##STR00287## ##STR00288## ##STR00289## ##STR00290## ##STR00291## ##STR00292## ##STR00293## ##STR00294## ##STR00295## ##STR00296## ##STR00297## ##STR00298## ##STR00299## ##STR00300## ##STR00301## and their pharmaceutically acceptable salts.

9. Compound of the following formula (I-0): ##STR00302## wherein: X represents NH, O, S or CH.sub.2; n represents an integer comprised from 3 to 7, or n being equal to 5; Y represents a group selected from: ##STR00303## Z representing O, S or NH; R representing: H a linear or branched (C.sub.1-C.sub.7)-alkyl, or methyl, ethyl, isopropyl or isobutyl, a group of formula (CH.sub.2).sub.iX(CH.sub.2).sub.jH, wherein X represents O, S or NH, i is an integer from 1 to 7, and j is an integer from 0 to 7, or a group CH.sub.2OCH.sub.3, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7)-cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, said aryl being an aromatic or heteroaromatic group, an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, CF.sub.3, adamantyl, CHRa-NH.sub.2, wherein Ra represents the side chain of a proteinogenic aminoacid, a cyclodextrin, or a cyclodextrin chosen from -cyclodextrin (-CD), -cyclodextrin (-CD), -cyclodextrin (-CD) and their derivatives, or alkylated -cyclodextrins, alkylated -cyclodextrins and alkylated -cyclodextrins, or a cyclodextrin of one the following formulae: ##STR00304## said (C.sub.1-C.sub.7)-alkyl, group of formula (CH.sub.2).sub.iX(CH.sub.2).sub.jH, (C.sub.2-C.sub.7)-alkenyl, (C.sub.2-C.sub.7)-alkynyl, (C.sub.3-C.sub.7)-cycloalkyl, (C.sub.5-C.sub.7)-cycloalkenyl, (C.sub.3-C.sub.7)-heterocycloalkyl, (C.sub.5-C.sub.7)-heterocycloalkenyl, CO-(C.sub.1-C.sub.7)-alkyl, CO.sub.2-(C.sub.1-C.sub.7)-alkyl, CONH-(C.sub.1-C.sub.7)-alkyl, aryl, alkyl aryl, CO-aryl and cyclodextrin being substituted or not by one or more substituent(s), each independently selected from: a linear or branched (C.sub.1-C.sub.7)-alkyl, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7)-cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, wherein the aryl is an aromatic or heteroaromatic group an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CHO, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, a halogen selected from the group comprising F, Cl, Br, and I, CF.sub.3, OR.sub.a, wherein R.sub.a represents: H, a linear or branched (C.sub.1-C.sub.7) -alkyl, a (C.sub.3-C.sub.7)-cycloalkyl, CO-(C.sub.1-C.sub.7)-alkyl, or CO-aryl, wherein aryl is an aromatic or heteroaromatic group, NR.sub.bR.sub.c, wherein R.sub.b and R.sub.c represent independently from each other: H, a linear or branched (C.sub.1-C.sub.7) -alkyl, a (C.sub.3-C.sub.7)-cycloalkyl, CO-(C.sub.1-C.sub.7)-alkyl, or CO-aryl, wherein aryl is an aromatic or heteroaromatic group, NO.sub.2, CN, SO.sub.3H or one of its salts, or SO.sub.3Na; and its pharmaceutically acceptable salts, provided that when R represents CHRa-NH.sub.2, then Y can only represent the following group (a): ##STR00305## with the proviso that said compound is not of the following structure: ##STR00306## and its salts.

10. Compound according to claim 9, of the following formula (I-1): ##STR00307## wherein: X represents NH, O, S or CH.sub.2; n represents an integer comprised from 3 to 7, or n being equal to 5; Y represents a group selected from: ##STR00308## Z representing O, S or NH; R.sub.1 representing: H a linear or branched (C.sub.1-C.sub.7)-alkyl, or isopropyl, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7)-cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, said aryl being an aromatic or heteroaromatic group, an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, CF.sub.3, adamantyl, CHRa-NH.sub.2, wherein Ra represents the side chain of a proteinogenic aminoacid, said (C.sub.1-C.sub.7) -alkyl, (C.sub.2-C.sub.7) -alkenyl, (C.sub.2-C.sub.7) -alkynyl, (C.sub.3-C.sub.7)-cycloalkyl, (C.sub.5-C.sub.7)-cycloalkenyl, (C.sub.3-C.sub.7)-heterocycloalkyl, (C.sub.5-C.sub.7) -heterocycloalkenyl, CO-(C.sub.1-C.sub.7)-alkyl, CO.sub.2-(C.sub.1-C.sub.7)-alkyl, CONH-(C.sub.1-C.sub.7)-alkyl, aryl, alkyl aryl and CO-aryl being substituted or not by one or more substituent(s), each independently selected from: a linear or branched (C.sub.1-C.sub.7)-alkyl, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7) -cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, wherein the aryl is an aromatic or heteroaromatic group an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CHO, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, a halogen selected from the group comprising F, Cl, Br, and I, CF.sub.3, OR.sub.a, wherein R.sub.a represents: H, a linear or branched (C.sub.1-C.sub.7) -alkyl, a (C.sub.3-C.sub.7) -cycloalkyl, CO-(C.sub.1-C.sub.7)-alkyl, or CO-aryl, wherein aryl is an aromatic or heteroaromatic group, NR.sub.bR.sub.c, wherein R.sub.b and R.sub.c represent independently from each other: H, a linear or branched (C.sub.1-C.sub.7) -alkyl, a (C.sub.3-C.sub.7) -cycloalkyl, CO-(C.sub.1-C.sub.7)-alkyl, or CO-aryl, wherein aryl is an aromatic or heteroaromatic group, NO.sub.2, CN; and its pharmaceutically acceptable salts, provided that when R.sub.1 represents CHRa-NH.sub.2, then Y can only represent the following group (a): ##STR00309##

11. Compound according to claim 9, wherein Y represents: ##STR00310## said compound being of following formula (I-1a): ##STR00311## X, n and R.sub.1 being as previously defined, or ##STR00312## said compound being of following formula (I-1b): ##STR00313## X, n and R.sub.1 being as defined, or ##STR00314## Z being as defined, said compound being of following formula (I-1c): ##STR00315## X, n, Z and R.sub.1 being as defined.

12. Process of preparation of a compound of formula (I-0): ##STR00316## wherein: X represents NH, O, S or CH.sub.2; n represents an integer being equal to 3, 4, 5, 6 or 7, or n being equal to 5; Y represents a group selected from: ##STR00317## Z representing O, S or NH; R representing: H a linear or branched (C.sub.1-C.sub.7) -alkyl, or methyl, ethyl, isopropyl or isobutyl, a group of formula (CH.sub.2).sub.iX(CH.sub.2).sub.jH, wherein X represents O, S or NH, i is an integer from 1 to 7, and j is an integer from 0 to 7, or a group CH.sub.2OCH.sub.3, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7)-cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, said aryl being an aromatic or heteroaromatic group, an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, CF.sub.3, adamantyl, CHRa-NH.sub.2, wherein Ra represents the side chain of a proteinogenic aminoacid, a cyclodextrin, or a cyclodextrin chosen from -cyclodextrin (-CD), -cyclodextrin (-CD), -cyclodextrin (-CD) and their derivatives, or alkylated -cyclodextrins, alkylated -cyclodextrins and alkylated -cyclodextrins, or a cyclodextrin of one of the following formulae: ##STR00318## said (C.sub.1-C.sub.7) -alkyl, group of formula (CH.sub.2).sub.iX(CH.sub.2).sub.jH, (C.sub.2-C.sub.7) -alkenyl, (C.sub.2-C.sub.7) -alkynyl, (C.sub.3-C.sub.7) -cycloalkyl, (C.sub.5-C.sub.7)-cycloalkenyl, (C.sub.3-C.sub.7)-heterocycloalkyl, (C.sub.5-C.sub.7)-heterocycloalkenyl, CO-(C.sub.1-C.sub.7)-alkyl, CO.sub.2-(C.sub.1-C.sub.7)-alkyl, CONH-(C.sub.1-C.sub.7)-alkyl, aryl, alkyl aryl, CO-aryl and cyclodextrin being substituted or not by one or more substituent(s), each independently selected from: a linear or branched (C.sub.1-C.sub.7)-alkyl, a linear or branched (C.sub.2-C.sub.7)-alkenyl, a linear or branched (C.sub.2-C.sub.7)-alkynyl, a (C.sub.3-C.sub.7)-cycloalkyl, a (C.sub.5-C.sub.7)-cycloalkenyl, a (C.sub.3-C.sub.7)-heterocycloalkyl, a (C.sub.5-C.sub.7)-heterocycloalkenyl, an aryl, wherein the aryl is an aromatic or heteroaromatic group an alkyl aryl, wherein the aryl is an aromatic or heteroaromatic group, a CHO, a CO-(C.sub.1-C.sub.7)-alkyl, a CO-aryl, wherein aryl is an aromatic or heteroaromatic group, a CO.sub.2H, a CO.sub.2-(C.sub.1-C.sub.7)-alkyl, a CONH-(C.sub.1-C.sub.7)-alkyl, a halogen selected from the group comprising F, Cl, Br, and I, CF.sub.3, OR.sub.a, wherein R.sub.a represents: H, a linear or branched (C.sub.1-C.sub.7) -alkyl, a (C.sub.3-C.sub.7)-cycloalkyl, CO-(C.sub.1-C.sub.7)-alkyl, or CO-aryl, wherein aryl is an aromatic or heteroaromatic group, NR.sub.bR.sub.c, wherein R.sub.b and R.sub.c represent independently from each other: H, a linear or branched (C.sub.1-C.sub.7) -alkyl, a (C.sub.3-C.sub.7)-cycloalkyl, CO-(C.sub.1-C.sub.7)-alkyl, or CO-aryl, wherein aryl is an aromatic or heteroaromatic group, NO.sub.2, CN, SO.sub.3H or one of its salts, or SO.sub.3Na; and its pharmaceutically acceptable salts, provided that when R represents CHRa-NH.sub.2, then Y can only represent the following group (a): ##STR00319## with the proviso that said compound is not of one of the following structures: ##STR00320## and its salts, comprising the following steps: when Y represents: ##STR00321## reaction between a compound of formula (1a): ##STR00322## wherein Rp represents an ad hoc hydroxyl protecting group, and a compound of formula (2a): ##STR00323## wherein R.sub.1 is a group R that is optionally protected by one or more ad hoc protecting groups, in presence of triphenylphosphine, a coupling agent and optionally 1-hydroxybenzotriazole (HOBt) or 1-hydroxy-7-aza-benzotriazole (HOAt), to obtain a compound of formula (3a): ##STR00324## cleavage of the Rp protecting groups and of the optional protecting groups of R.sub.1 in said compound of formula (3a), to obtain a compound of formula (I-0) wherein Y represents (a), of following formula (I-0a): ##STR00325## when Y represents: ##STR00326## reaction between a compound of formula (1b): ##STR00327## (1b), and a compound of formula (2b): ##STR00328## wherein R.sub.1 is a group R that is optionally protected by one or more ad hoc protecting groups, to obtain a compound of formula (3b): ##STR00329## cleavage of the Rp protecting groups and of the optional protecting groups of R.sub.1 in said compound of formula (3b), to obtain a compound of formula (I-0) wherein Y represents (b), of following formula (I-0b) ##STR00330## when Y represents: ##STR00331## reaction between a compound of formula (1c): ##STR00332## and a compound of formula (2b): ##STR00333## wherein R.sub.1 is a group R that is optionally protected by one or more ad hoc protecting groups, to obtain a compound of formula (3b): ##STR00334## cleavage of the Rp protecting groups and of the optional protecting groups of R.sub.1 in said compound of formula (3c), to obtain a compound of formula (I-0) wherein Y represents (c), of following formula (I-0c): ##STR00335##

Description

DESCRIPTION OF THE DRAWINGS

[1024] FIG. 1 presents the adhesion levels of Adherent-Invasive E. coli strain LF82 to intestinal epithelial cells T84 in the presence of increasing doses of compounds 5 and 10 or D-mannose, using co-, pre- and post-incubation protocols (respectively FIGS. 1A, 1B and 1C). Results are expressed in percentages of residual adhesion, considering 100% as the LF82 adhesion level in absence of any compound (meanssem). *: p<0.05; **:p<0.01; ***: p<0.001 (t test).

[1025] FIG. 2 presents the adhesion levels of Adherent-Invasive E. coli strain LF82 to colonic mucosa from CEABAC10 mice, in presence of compound 10 at a concentration of 100 M. Infections were performed with 100 L of a bacterial suspension of AIEC LF82 at 2.510.sup.7 bacteria/mL. Results are expressed in CFU/g of tissue, each point represents result for LF82 adhesion in one colonic loop (horizontal bars=median).

[1026] FIG. 3 presents the body weight of CEABAC10 mice infected with 10.sup.9 AIEC LF82 bacteria at day 0, after two oral administrations of the monovalent compounds 5 and 10 at a dose of 10 mg/kg. Each point represents the meansem of body weights for each group of mice. Results are expressed as percentages, day 0 (LF82 infection) being considered as 100%. NI: non-infected. a: p<0.001; b: p<0.01 compared to LF82-infected mice (t test).

[1027] FIG. 4 presents the bacterial colonization in feces (FIGS. 4A, 4B and 4C) and Disease Activity Index score (DAI) (D) at respectively day 1, 3 and 4 post-infection of CEABAC10 mice infected with 10.sup.9 AIEC LF82 at day 0. Two oral administrations of monovalent compounds 5 and 10 were realized at a dose of 10 mg/kg. For FIGS. 4A to 4C, each point represents the number of colony forming units (CFU) of AIEC LF82 per gram of feces for each mouse. Horizontal red bars represent medians. FIG. 4D presents DAI scores, expressed as meanssem. NI: non-infected. *: p<0.05; **: p<0.01; ***:p<0.001 (t test, in comparison with LF82 group).

[1028] FIG. 5 presents the assessment of bacteria-associated to the ileum (FIG. 5A) and to the colon (FIG. 5B) of CEABAC10 mice infected with AIEC LF82 after oral treatment with monovalent compounds 5 and 10 (day 4 post-infection). Mice were orally challenged with 10.sup.9 bacteria at day 0 (DO) and monovalent compounds were administrated two times at a dose of 10 mg/kg. Each point represents the number of colony forming units (CFU) of AIEC LF82 per gram of feces for each mouse, horizontal bars represent medians. NI: non-infected mice.

[1029] FIG. 6 presents the weight of spleens of AIEC LF82-infected CEABAC10 mice after oral treatment with monovalent compounds 5 and 10 at day 4 post-infection. Mice were orally challenged with 10.sup.9 bacteria at day 0 (DO) and monovalent compounds were administrated two times at a dose of 10 mg/kg. Results are expressed as meanssem. NI: non-infected. *: p<0.05 (t test).

[1030] FIG. 7 presents the contents in pro-inflammatory cytokines KC (FIG. 7A), TNF- (FIG. 7B) and IL-23 (FIG. 7C) secreted by colonic mucosa at day 4 post-infection of CEABAC10 mice infected with AIEC LF82 after oral treatment with monovalent compounds 5 and 10. Mice were orally challenged with 10.sup.9 bacteria at day 0 (DO) and monovalent compounds were administrated two times at a dose of 10 mg/kg. Each point represents the level of secreted cytokines in pg/mL, for one mouse. Horizontal bars represent medians. *: p<0.05 (t test).

[1031] FIG. 8 presents the residual adhesion of LF82 bacteria on T84 cells in a post-incubation assay. Results are expressed in percentage of residual considering 100% as adhesion of LF82 AIEC (infected control) without any inhibitory compound (tested molecule). Different concentrations of anti-FimH molecules are tested: 0.1, 1, 5, 10, 50 and 100 M. T test=Student: a: p<0.05, b: p<0.01, c: p<0.001.

[1032] FIG. 9A presents the body weight of CEABAC10 transgenic mice uninfected or infected with MEC LF82 measured at D=1, D=2, D=3 and D=4 days post infection. Non-infected=Non infected mice (negative control); LF82 =Infected mice with the LF82 AIEC strain (positive control); LF82+10, LF82+5=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg.

[1033] FIG. 9B presents the body weight of CEABAC10 transgenic mice uninfected or infected with AlEC LF82 measured at D=1, D=2, D=3 and D=4 days post infection. Non-infected =Non infected mice (negative control); LF82 =Infected mice with the LF82 AIEC strain (positive control); LF82+16, LF82+22=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg.

[1034] FIG. 10 presents the DAI (Disease Activity Index) measured at D=3 days post infection. LF82=Infected mice with the LF82 AIEC strain (positive control); LF82+16, LF82+22, LF82+10, LF82+5=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg.

[1035] FIG. 11 presents the colony forming units (CFU) measured per gram of feces at D=1 and D=3 days. NI=Non infected mice (negative control); LF82=Infected mice with the LF82 AIEC strain (positive control); LF82+16, LF82+22=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg. Median value is indicated in horizontal line.

[1036] FIG. 12 presents the colony forming units (CFU) measured per gram of ileum and colon (mucosa) after D=3 days. LF82=Infected mice with the LF82 AIEC strain (positive control); LF82+16, LF82+22=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg. Median value is indicated in horizontal line.

[1037] FIG. 13 presents the Myelopexoridase activity (MPO) assessment in intestinal tissue from CEABAC10 mice measured in ng/mL. LF82=Infected mice with the LF82 AIEC strain (positive control); LF82+10, LF82+5, LF82+16, LF82+22=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg. Median value is indicated in horizontal line. T test=Student: *p<0.05, **P<0.01, ***p<0.001.

[1038] FIG. 14A presents the IL-23 assessment in blood from CEABAC10 mice measured in pg/mL. Non-infected=non infected mice (negative control); LF82=Infected mice with the LF82 AIEC strain (positive control); LF82+10, LF82+5=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg. Median value is indicated in horizontal line. T test=Student: *p<0.05, **P<0.01, ***p<0.001.

[1039] FIG. 14B presents the IL-23 assessment in blood from CEABAC10 mice measured in pg/mL. Non-infected=non infected mice (negative control); LF82=Infected mice with the LF82 AIEC strain (positive control); LF82+16, LF82+22=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg. Median value is indicated in horizontal line. T test=Student: *p<0.05, **P<0.01, ***p<0.001.

[1040] FIG. 15A presents the IL-1beta assessment in blood from CEABAC10 mice measured in pg/mL. Non-infected=non infected mice (negative control); LF82=Infected mice with the LF82 AIEC strain (positive control); LF82+10, LF82+5=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg. Median value is indicated in horizontal line. T test=Student: *p<0.05, **P<0.01, ***p<0.001

[1041] FIG. 15B presents the IL-1beta assessment in blood from CEABAC10 mice measured in pg/mL. Non-infected=non infected mice (negative control); LF82=Infected mice with the LF82 AIEC strain (positive control); LF82+16, LF82+DA22=Infected mice with the LF82 AIEC strain treated with molecules tested at 10 mg/kg. Median value is indicated in horizontal line. T test=Student: *p<0.05, **P<0.01, ***p<0.001.

EXAMPLES

[1042] A. Synthesis of monovalent heptylmannoside cyclodextrin compounds

Example 1

Synthesis of Compound 5

[1043] ##STR00257##

[1044] 8-Oxaundec-10-ynyl 2,3,4,6-tetra-O-acetyl--D-mannopyranoside 3

[1045] Mannosyl pentaacetate 1 (229 mg, 0.587 mmol), compound 2 (150 mg, 0.882 mmol) and silver trifluoroacetate (194 mg, 0.878 mmol) were dissolved in dry dichloromethane (3 mL). A solution of SnCl.sub.4 1M in dichloromethane (585 L) was added and the mixture was stirred at rt for 3 h under argon atmosphere. The solution was diluted in dichloromethane (10 mL) and washed with NaHCO.sub.3 satd. (210 mL). The organic layer was dried, filtered and evaporated under reduced pressure. The residue was chromatographied on silica gel with ethyl acetate-cyclohexane (2-8) to (3-7) to afford 3 as a colorless oil (128 mg, 44%). Analytical data were identical as previously described [Gouin, S. G.; Wellens, A.; Bouckaert, J.; Kovensky, J. ChemMedChem. 2009, 5, 749-755].

[1046] 8-Oxaundec-10-ynyl--D-mannopyranoside 4

3 (400 mg, 800 mop was dissolved in MeOH (10 mL). A solution of freshly prepared sodium methanolate 1M in methanol (500 L) was added and the mixture was stirred at rt for 4 h. Amberlyst IR120 (H.sup.+) was added and the mixture stirred until pH reached 5. The resin was filtered off and the solution was evaporated to dryness leading to unprotected product 4 (263 mg, 99%).

[1047] [].sub.D=+96 (c=0.2, MeOH); .sup.1H NMR (300 MHz, CD.sub.3OD) =4.76 (1 H, d, J=1.6 Hz, H-1), 4.14 (2 H, d, J=2.4 Hz, OCH.sub.2C), 3.82-3.80 (2 H, m, H-2, H-3), 3.75-3.69 (3 H, m, H-5, 2H-6), 3.64 (1 H, t, J=9.3 Hz, H-4), 2.84 (1 H, t, CCH), 1.61-1.55 (4 H, br, 2CH.sub.2), 1.39 (6 H, br, 6CH.sub.2); .sup.13C NMR (125 MHz, D.sub.2O): =102.4 (C1), 76.5 (CCH), 75.5, 73.5, 73.1, 71.8 (C-2, -3,-4,-5), 69.4 (CH.sub.2O), 59.6 (CH.sub.2CCH), 31.4, 31.3, 31.1, 28.1, 28.0 (CH.sub.2); HRMS (ES+): Found 355.1732 C.sub.16H.sub.28O.sub.7Na requires 355.1733.

[1048] compound 5

##STR00258##

[1049] Alkynyl-saccharide 4 (29 mg, 87 mol) and mono-6-azido-6-deoxy-beta-cyclodextrin (50 mg, 43 mol) were dissolved in a DMF/H.sub.2O mixture (2/0.5 mL). Copper sulfate (6.9 mg, 43 mol) and sodium ascorbate (17 mg, 86 mol) were added and the mixture was stirred at 70 C. for 30 minutes under W irradiation. Ethylenediamine tetraacetic acid trisodium salt (50 mg, 127 mol) was added and the mixture was stirred for 10 minutes at rt. The mixture was evaporated under reduced pressure and the residue purified by preparative HPLC leading to compound 5 (33 mg, 51%) as a white powder after lyophilisation.

[1050] [].sub.D=+130 (c=0.1, MeOH); Tr=17 min; .sup.1H NMR (500 MHz, D.sub.2O) =8.23 (1 H, s, H.sub.triazol), 5.51, 5.36, 5.30 (7 H, 3s, H-1.sup.I-VII), 5.15 (1 H, s, H-1.sup.HM), 4.20-3.20 (54 H, br, H-2,-3,-4,-5,.sup.I-VII, H-2,-3,-4,-5,-6.sup.HM, O-CH.sub.2-triazol, 2CH.sub.2), 1.72, 1.65, 1.47 (10 H, br, (CH.sub.2), .sup.13C NMR (125 MHz, D.sub.2O): =146.1 (C=CH.sub.triazol), 123.8 (CH=C.sub.triazol), 102.1, 101.8, 99.9 (C1.sup.I-VII, C1.sup.HM), 83.1, 81.7, 80.9, 80.3 (C4.sup.I-VII), 72.1, 71.0, 70.4, 68.6, 67.0, 66.5, 63.0, 60.7, 59.8, 58.8 (C2,-3,-4,-5,-6.sup.HM, CH.sub.2O), 51.5 (C6.sup.I), 29.1, 28.5, 28.0, 25.7, 25.1 (CH.sub.2); HRMS (ES+): Found 1514.5564 C.sub.58H.sub.97N.sub.3O.sub.41Na requires 1514.5495.

B. Synthesis of mannosyl-O-heptylamides

##STR00259##

[1051] a) Carboxylic acid, HOBt, DIC, PH.sub.3P, THF, 0 C..fwdarw.rt

General Procedure A: One Pot Staudinger-Amide Coupling:

[1052] The azide-functionalized carbohydrate (1 equiv.) and the carboxylic acid (1.8 equiv.) were combined with HOBt (1.8 equiv.) in a flask and dried for more than 1 h in vacuo. This mixture was dissolved in dry THF (25 mL/mmol azide) under nitrogen and cooled to 0 C. Then DIC (1.8 equiv.) was added and the solution was stirred for 10 min, followed by the addition of Ph.sub.3P (1.8 equiv.) and stirring for 1 h at 0 C. Then the reaction mixture was stirred overnight at room temperature, diluted with water (50 mL) and extracted twice with ethyl acetate (30 mL). The combined organic phases were washed with brine, dried with MgSO.sub.4 and the mixture was filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography.

Example 2

Compound 7

[1053] According to the general procedure A, mannosyl azide 6 (50 mg, 0.099 mmol), isobutyric acid (16 mg, 0.178 mmol, 1.8 equiv.), HOBt (24 mg, 0.178 mmol, 1.8 equiv.), DIC (28 L, 0.178 mmol, 1.8 equiv.) and Ph.sub.3P (47 mg, 0.178 mmol, 1.8 equiv.) were allowed to react in THF (2.5 mL). The crude product was purified by silica gel column chromatography (EtOAc/petroleum ether, 70:30.fwdarw.EtOAc as eluents) to give the amide 7 (42 mg, 0.079 mmol, 80%) as an oil.

[1054] [].sub.D=+71 (c=0.81 in CHCl.sub.3)

[1055] .sup.1H NMR (300 MHz, CDCl.sub.3): =1.12 (6H, d, J=6.9 Hz, 2CH.sub.3-isobutyric acid), 1.28-1.59 (10H, m), 1.97 (3H, s, AcO), 2.02 (3H, s, AcO), 2.08 (3H, s, AcO), 2.13 (3H, s, AcO), 2.31 (1H, m, CH, isobutyric acid), 3.21 (2H, m, H-7), 3.41 (1H, m, H-1a), 3.65 (1H, m, H-1b), 3.95 (1H, ddd, J.sub.5,4=9.5 Hz, J.sub.5,6b=5.3 Hz, J.sub.5,6a=2.5 Hz, H-5), 4.10 (1H, dd, J.sub.6a,6b=12.2 Hz, J.sub.6a,5=2.5 Hz, H-6a), 4.25 (1H, dd, J.sub.6b,6a=12.2 Hz, J.sub.6b,5=5.3 Hz, H-6a), 4.77 (1H, d, J.sub.1,2=1.7 Hz, H-1), 5.20 (1H, dd, J.sub.2,3=3.3 Hz, J.sub.2,1=1.7 Hz, H-2), 5.24 (1H, dd, J.sub.4,3=10.1 Hz, J.sub.4,5=9.6 Hz, H-4), 5.32 (1H, dd, J.sub.3,4=10.1 Hz, J.sub.3,2=3.3 Hz, H-3), 5.60 (1H, bs, NH).

[1056] .sup.13C NMR (100.6 MHz, CDCl.sub.3): =19.6 (2CH.sub.3, isobutyric acid), 20.6 (2CH.sub.3, 2AcO), 20.7 (CH.sub.3, AcO), 20.8 (CH.sub.3, AcO), 25.8 (CH.sub.2), 26.6 (CH.sub.2), 28.8 (CH.sub.2), 29.0 (CH.sub.2), 29.5 (CH.sub.2), 35.6 (CH, isobutyric acid), 39.2 (CH.sub.2, C-7), 62.5 (CH, C-6), 66.2 (CH), 68.3 (CH, CH.sub.2, C-5, C-1), 69.1 (CH), 69.6 (CH, C-2), 97.5 (CH, C-1), 169.7 (C, AcO), 169.9 (C, AcO), 170.1 (C, AcO), 170.6 (C, AcO), 176.8 (C, amide).

[1057] MS (CI, NH.sub.3): m/z 549 [M+NH.sub.3]1.sup.+.

Example 3

Compound 8

[1058] According to the general procedure A, mannosyl azide 6 (50 mg, 0.099 mmol), N-Boc-L-alanine (34 mg, 0.178 mmol, 1.8 equiv.), HOBt (24 mg, 0.178 mmol, 1.8 equiv.), DIC (28 L, 0.178 mmol, 1.8 equiv.) and Ph.sub.3P (47 mg, 0.178 mmol, 1.8 equiv.) were allowed to react in THF (2.5 mL). The crude product was purified by silica gel column chromatography (EtOAc/petroleum ether, 70:30.fwdarw.EtOAc as eluents) to give the amide 8 (35 mg, 0.055 mmol, 56%) as an oil.

[1059] [].sub.D=+54 (c=0.92 in CHCl.sub.3)

[1060] .sup.1H NMR (300 MHz, CDCl.sub.3): =1.33 (3H, d, J=7.0 Hz, CH.sub.3-alanine), 1.25-1.76 (10H, m), 1.44 (3H, s, N-Boc), 2.00 (3H, s, AcO), 2.05 (3H, s, AcO), 2.10 (3H, s, AcO), 2.16 (3H, s, AcO), 3.24 (2H, t, J=6.6 Hz, H-7), 3.43 (1H, m, H-1a), 3.67 (1H, m, H-1b), 3.97 (1H, ddd, J.sub.5,4=8.2 Hz, J.sub.5,6b=5.3 Hz, J.sub.5,6a=2.5 Hz, H-5), 4.11 (1H, dd, J.sub.6a,6b=12.1 Hz, J.sub.6a,5=2.4 Hz, H-6a), 4.12 (1H, m , CH-alanine), 4.28 (1H, dd, J.sub.6b,6a=12.1 Hz, J.sub.6b,5=5.3 Hz, H-6a), 4.79 (1H, d, J.sub.1,2=1.6 Hz, H-1), 5.03 (1H, bs, NH), 5.22 (1H, dd, J.sub.2,3=3.1 Hz, J.sub.2,1=1.6 Hz, H-2), 5.27 (1H, dd, J.sub.4,3=10.0 Hz, J.sub.4,5=8.2 Hz, H-4), 5.34 (1H, dd, J.sub.3,4=10.0 Hz, J.sub.3,2=3.3 Hz, H-3), 6.19 (1H, bs, NH).

[1061] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =18.4 (CH.sub.3, alanine), 20.7 (CH.sub.3, AcO), 20.72 (2 CH.sub.3, AcO), 20.9 (CH.sub.3, AcO), 25.8 (CH.sub.2), 26.5 (CH.sub.2), 28.3 (3 X CH.sub.3, N-Boc), 28.8 (CH.sub.2), 29.0 (CH.sub.2), 29.3 (CH.sub.2), 39.4 (CH.sub.2, C-7), 50.1 (C, N-Boc), 62.5 (CH, C-6), 66.2 (CH, C-1), 68.4 (2 CH), 69.1 (CH), 69.7 (CH, C-2), 97.5 (CH, C-1), 155.5 (C, amide), 169.7 (C, AcO), 170.0 (C, AcO), 170.1 (C, AcO), 170.6 (C, AcO), 172.4 (C, N-Boc).

[1062] MS (CI, NH3): m/z 633 [M]+

[1063] HRMS (MALDI, DHB): m/z calcd for C29H48N2O13Na [M+Na]+: 655.3049, found: 655.3026.

Example 4

Compound 9

[1064] According to the general procedure A, mannosyl azide 6 (50 mg, 0.099 mmol), picolinic acid (22 mg, 0.178 mmol, 1.8 equiv.), HOBt (24 mg, 0.178 mmol, 1.8 equiv.), DIC (28 L, 0.178 mmol, 1.8 equiv.) and Ph.sub.3P (47 mg, 0.178 mmol, 1.8 equiv.) were allowed to react in THF (2.5 mL). The crude product was purified by silica gel column chromatography (EtOAc/petroleum ether, 70:30.fwdarw.EtOAc as eluents) to give the amide 9 (43 mg, 0.076 mmol, 77%) as an oil.

[1065] [].sub.D=+61 (c=1.03 in CHCl.sub.3)

[1066] .sup.1H NMR (300 MHz, CDCl.sub.3): 1.30-1.69 (10H, m), 1.98 (3H, s, AcO), 2.03 (3H, s, AcO), 2.09 (3H, s, AcO), 2.14 (3H, s, AcO), 3.39 (1H, m, H-1a), 3.46 (2H, q, J=6.8 Hz, H-7), 3.66 (1H, m, H-1b), 3.97 (1H, ddd, J.sub.5,4=9.5 Hz, J.sub.5,6b=5.2 Hz, J.sub.5,6a=2.3 Hz, H-5), 4.09 (1H, dd, J.sub.6a,6b=12.2 Hz, J.sub.6a,5=2.3 Hz, H-6a), 4.28 (1H, dd, J.sub.6b,6a=12.2 Hz, J.sub.6b,5=5.2 Hz, H-6a), 4.79 (1H, d, J.sub.1,2=1.6 Hz, H-1), 5.22 (1H, dd, J.sub.2,3=3.3 Hz, J.sub.2,1=1.6 Hz, H-2), 5.26 (1H, dd, J.sub.4,3=9.8 Hz, J.sub.4,5=9.6 Hz, H-4), 5.34 (1H, dd, J.sub.3,4=9.8 Hz, J.sub.3,2=3.3 Hz, H-3), 7.41 (1H, ddd, J=7.6 Hz, J=4.8 Hz, J=1.3 Hz, picolinic), 7.84 (1H, ddd, J=7.6 Hz, J=7.6 Hz, J=1.7 Hz, picolinic), 8.08 (1H, bs, NH), 8.19 (1H, bd, J=7.8 Hz, picolinic), 8.54 (1H, ddd, J=4.7 Hz, J=1.7 Hz, J=0.9 Hz, picolinic).

[1067] .sup.13C NMR (100.6 MHz, CDCl.sub.3): =20.7 (CH.sub.3, AcO), 20.71 (2 X CH.sub.3, AcO), 20.9 (CH.sub.3, AcO), 26.0 (CH.sub.2), 26.8 (CH.sub.2), 29.0 (CH.sub.2), 29.1 (CH.sub.2), 29.5 (CH.sub.2), 39.4 (CH.sub.2, C-7), 62.5 (CH, C-6), 66.2 (CH, C-1), 68.3 (CH), 68.4 (CH), 69.1 (CH), 69.7 (CH, C-2), 97.5 (CH, C-1), 122.3 (CH, picolinic acid), 126.1 (CH, picolinic acid), 137.5 (CH, picolinic acid), 147.8 (CH, picolinic acid), 149.9 (C, picolinic acid), 164.0 (C, amide), 169.7 (C, AcO), 169.9 (C, AcO), 170.1 (C, AcO), 170.6 (C, AcO).

[1068] MS (CI, NH3): m/z (%): 567 [M]+

[1069] HRMS (MALDI, DHB): m/z calcd for C27H38N2O11Na [M+Na]+: 598.2368, found: 589.2374.

##STR00260##

i. NaOMe, MeOH, rt, ii. Amberlite IR120 (H), iii. TFA-DCM, 0 C. vi. HCl ac. (for N-Boc protected compound 8).

General Procedure B: O-Acetyl Deprotection According to Zempln Conditions

[1070] The protected glycosyl amide (1 equiv.) was dissolved in dry MeOH (30 mL) and sodium methoxide (1 M solution in MeOH, 10% per AcO) was added. The mixture was stirred for 4 h, neutralized with Amberlite IR120 (H), filtered and the solvents evaporated to dryness. The substrate was dissolved in water and subjected to lyophilization.

General Procedure C: N-Boc Deprotection with Trifluoroacetic Acid

[1071] The Boc-protected amine was dissolved in DCM (2 mL/mmol) and TFA (2 mL/mmol) was added at 0 C. The mixture was stirred for 1 h, evaporated to dryness and co-evaporated with H.sub.2O (3 times) and 0.5 N HCl (3 times). The substrate was dissolved in water and subjected to lyophilization.

Example 5

Compound 10

[1072] According to the general procedure B, using the amide 7 (81 mg, 0.128 mmol) as starting material, the derivative 10 was obtained after lyophilization (41 mg, 0.113 mmol, 93%), as an amorphous white solid.

[1073] [].sub.D=+48.1 (c=1.32 in MeOD).

[1074] .sup.1H NMR (300 MHz, MeOD): =1.10 (6H, d, J=6.9 Hz, 2CH.sub.3-isobutyric acid), 1.29-1.64 (10H, m), 2.42 (1H, m, CH-isobutyric acid), 3.15 (2H, t, J=6.9 Hz, C-7), 3.41 (1H, m, H-1a), 3.52 (1H, ddd, J.sub.5,4=9.2 Hz, J.sub.5,6b=5.6 Hz, J.sub.5,6a=2.4 Hz, H-5), 3.61 (1H, dd, J.sub.4,3=9.4 Hz, J.sub.4,5=9.2 Hz, H-4), 3.67-3.75 (3H, m), 3.78 (1H, dd, J.sub.2,3=3.3 Hz, J.sub.2,1=1.7 Hz, H-2), 3.82 (1H, dd, J.sub.6b,6a=11.9 Hz, J.sub.6b,5=2.5 Hz, H-6b), 4.73 (1H, d, J.sub.1,2=1.7 Hz, H-1).

[1075] .sup.13C NMR (100.6 MHz, MeOD): =20.0 (2CH.sub.3, 2CH.sub.3-isobutyric acid), 27.2 (CH.sub.2), 27.8 (CH.sub.2), 30.1 (CH.sub.2), 30.4 (CH.sub.2), 30.5 (CH.sub.2), 36.3 (CH, CH-isobutyric acid), 40.2 (CH.sub.2, C-7), 62.9 (CH, C-6), 68.5 (CH), 68.6 (CH), 72.2 (CH, C-5), 72.6 (CH, C-1), 74.6 (CH, C-2), 101.5 (CH, C-1), 180.0 (C, amide).

[1076] MS (CI, NH.sub.3): m/z 364 [M+H].sup.+

Example 6

Compound 11

[1077] According to the general procedure B and C, using the amide 8 (81 mg, 0.128 mmol) as starting material, the alanine derivative 11 was obtained after lyophilization (41 mg, 0.102 mmol, 80%), in form of ammonium chloride salt, as an amorphous white solid.

[1078] [].sub.D=+39.6 (c=1.21 in D.sub.2O).

[1079] .sup.1H NMR (300 MHz, D.sub.2O): =1.12-1.48 (10H, m), 1.36 (3H, d, J=7.1 Hz, CH.sub.3-alanine), 3.08 (2H, m, H-1), 3.33-3.77 (8H, m), 3.87 (2H, q, J=7.1 Hz, CH-alanine), 4.67 (1H, bs, H-1).

[1080] .sup.13C NMR(100.6 MHz, D.sub.2O): =16.6 (CH.sub.3, alanine), 24.9 (CH.sub.2), 25.2 (CH.sub.2), 25.9 (CH.sub.2), 28.0 (CH.sub.2), 31.2 (CH.sub.2), 39.5 (CH.sub.2, C-7), 60.9 (CH, C-6), 61.8 (CH, C-1), 66.8 (CH, alanine), 67.9 (CH, C-5), 70.1 (CH), 70.7 (CH), 72.7 (CH, C-2), 99.7 (CH, C-1), 170.5 (C, amide).

[1081] HRMS (MALDI, DHB): m/z calcd for C.sub.16H.sub.32N.sub.2O.sub.7Na [M+Na]+: 387.2107, found: 387.2119

Example 7

Compound 12

[1082] According to the general procedure B, using the amide 9 (25 mg, 0.048 mmol) as starting material, the picolinic derivative 12 was obtained after lyophilization (19 mg, 0.048 mmol, quantitative) as an amorphous white solid.

[1083] [].sub.D=+47.1 (c=1.81 in MeOH)

[1084] .sup.1H NMR (300 MHz, MeOD): 1.34-1.68 (10H, m), 3.40 (1H, m, H-1a), 3.42 (2H, t, J=7.0 Hz, H-7), 3.52 (1H, ddd, J.sub.5,4=9.4 Hz, J.sub.5,6b=5.4 Hz, J.sub.5,6a=2.4 Hz, H-5), 3.62 (1H, dd, J.sub.4,3=9.4 Hz, J.sub.4,5=9.2 Hz, H-4), 3.68-3.76 (3H, m, H-3, H-6a, H-1b), 3.78 (1H, dd, J.sub.2,3=3.2 Hz, J.sub.2,1=1.6 Hz, H-2), 3.82 (1H, dd, J.sub.6b,6a=11.8 Hz, J.sub.6b,5=2.4 Hz, H-6b), 4.73 (1H, d, J.sub.1,2=1.6 Hz, H-1), 7.41 (1H, ddd, J=7.6 Hz, J=4.8 Hz, J=1.3 Hz, picolinic), 7.53 (1H, ddd, J=7.6 Hz, J=4.8 Hz, J=1.3 Hz, picolinic), 7.95 (1H, ddd, J=7.7 Hz, J=7.7 Hz, J=1.7 Hz, picolinic), 8.08 (1H, ddd, J=7.8 Hz, J=1.1 Hz, J=1.1 Hz, picolinic), 8.68 (1H, ddd, J=4.7 Hz, J=1.7 Hz, J=1.1 Hz, picolinic).

[1085] .sup.13C NMR(100.6 MHz, MeOD): =23.5 (CH.sub.2), 27.3 (CH.sub.2), 28.0 (CH.sub.2), 30.2 (CH.sub.2), 30.5 (CH.sub.2), 40.4 (CH.sub.2, C-7), 62.9 (CH, C-6), 68.5 (CH, C-1), 68.6 (CH), 72.2 (CH), 72.6 (CH), 74.5 (CH, C-2), 101.5 (CH, C-1), 123.0 (CH, picolinic acid), 127.6 (CH, picolinic acid), 138.7 (CH, picolinic acid), 149.7 (CH, picolinic acid), 151.1 (C, picolinic acid), 166.6 (C, amide).

[1086] MS (CI, NH.sub.3): m/z 399 [M+H].sup.+

[1087] HRMS (MALDI, DHB): m/z calcd for C42H48O5 [M+Na]+: 421.1945, found: 421.1965.

##STR00261##

[1088] a) Carboxylic acid, HOBt, DIC, PH.sub.3P, THF, 0 C..fwdarw.rt

Example 8

Compound 1:

[1089] According to the general procedure A, mannosyl azide 6 (100 mg, 0.205 mmol), n-butyric acid (32 L, 0.369 mmol, 1.8 equiv.), HOBt (50 mg, 0.369 mmol, 1.8 equiv.), DIC (57 L, 0.369 mmol, 1.8 equiv.) and Ph.sub.3P (97 mg, 0.369 mmol, 1.8 equiv.) were allowed to react in DCM (5 mL). The crude product was purified by silica gel column chromatography (EtOAc/diethyl ether, 80:20 as eluents) to give the amide 1 (47 mg, 0.088 mmol, 43%) as an oil.

[1090] [].sub.D=+37.5 (c=0.62 in CHCl.sub.3)

[1091] .sup.1H NMR (300 MHz, CDCl.sub.3): =0.94 (3H, t, J=7.4 Hz, CH.sub.3-butyric acid), 1.28-1.61 (10H, m), 1.66 (2H, m, CH.sub.2-butyric acid), 1.99 (3H, s, AcO), 2.04 (3H, s, AcO), 2.10 (3H, s, AcO), 2.14 (2H, m, CH.sub.2-butyric acid), 2.15 (3H, s, AcO), 3.24 (2H, m, H-7), 3.44 (1H, m, H-1a), 3.67 (1H, m, H-1b), 3.97 (1H, ddd, J.sub.5,4=9.5 Hz, J.sub.5,6b=5.4 Hz, J.sub.5,6a=2.5 Hz, H-5), 4.11 (1H, dd, J.sub.6a,6b=12.3 Hz, J.sub.6a,5=2.5 Hz, H-6a), 4.28 (1H, dd, J.sub.6b,6a=12.3 Hz, J.sub.6b,5=5.4 Hz, H-6a), 4.79 (1H, d, J.sub.1,2=1.6 Hz, H-1), 5.22 (1H, dd, J.sub.2,3=3.2 Hz, J.sub.2,1=1.7 Hz, H-2), 5.27 (1H, dd, J.sub.4,3=10.1 Hz, J.sub.4,5=9.6 Hz, H-4), 5.34 (1H, dd, J.sub.3,4=10.1 Hz, J.sub.3,2=3.3 Hz, H-3), 5.55 (1H, bs, NH).

[1092] .sup.13C NMR (100.6 MHz, CDCl.sub.3): =13.7 (CH.sub.3, butyric acid), 19.1 (CH.sub.2, butyric acid), 20.61 (2CH.sub.3, 2AcO), 20.66 (CH.sub.3, AcO), 20.8 (CH.sub.3, AcO), 25.8 (CH.sub.2), 26.6 (CH.sub.2), 28.8 (CH.sub.2), 29.0 (CH.sub.2), 29.4 (CH.sub.2), 38.7 (CH.sub.2, butyric acid), 39.3 (CH.sub.2, C-7), 62.4 (CH, C-6), 66.1 (CH), 68.3 (CH, CH.sub.2, C-5, C-1), 69.1 (CH), 69.6 (CH, C-2), 97.4 (CH, C-1), 169.7 (C, AcO), 169.9 (C, AcO), 170.1 (C, AcO), 170.6 (C, AcO), 172.9 (C, amide).

[1093] MS (CI, NH.sub.3): m/z 532 [M+H].sup.+

[1094] HRMS (ESI): m/z calcd for C.sub.25H.sub.41NO.sub.11Na[M+Na].sup.+: 554.2577, found: 554.2571.

Example 9

Compound 2

[1095] According to the general procedure A, mannosyl azide 6 (100 mg, 0.205 mmol), O-acetylactic acid (81 mg, 0.615 mmol, 3 equiv.), HOBt (83 mg, 0.615 mmol, 3 equiv.), DIC (95 L, 0.615 mmol, 3 equiv.) and Ph.sub.3P (97 mg, 0.615 mmol, 3 equiv.) were allowed to react in DCM (5.1 mL). The crude product was purified by silica gel column chromatography (EtOAc/petroleum ether, 80:20.fwdarw.EtOAc as eluents) to give the amide 2 (72 mg, 0.125 mmol, 61%) as a colorless oil.

[1096] [].sub.D=+53.9 (c=0.52 in CHCl.sub.3)

[1097] .sup.1H NMR (300 MHz, CDCl.sub.3): =1.26-1.34 (8H, m), 1.42 (3H, d, J=6.9 Hz, CH.sub.3-lactic acid), 1.52 (2H, m), 1.95 (3H, s, AcO), 2.00 (3H, s, AcO), 2.06 (3H, s, AcO), 2.10 (3H, s, AcO), 2.12 (3H, s, AcO), 3.23 (2H, q, J=6.7 Hz, H-7), 3.40 (1H, m, H-1a), 3.64 (1H, m, H-1b), 3.93 (1H, ddd, J.sub.5,4=9.3 Hz, J.sub.5,6b=5.2 Hz, J.sub.5,6a=2.3 Hz, H-5), 4.07 (1H, dd, J.sub.6a,6b=12.3 Hz, J.sub.6a,5=2.4 Hz, H-6a), 4.24 (1H, dd, J.sub.6b,6a=12.3 Hz, J.sub.6b,5=5.3 Hz, H-6a), 4.76 (1H, d, J.sub.1,2=1.6 Hz, H-1), 5.13 (1H, q, J=6.7 Hz, CH-lactic acid), 5.18 (1H, dd, J.sub.2,3=3.2 Hz, J.sub.2,1=1.6 Hz, H-2), 5.23 (1H, dd, J.sub.4,3=10.0 Hz, J.sub.4,5=9.3 Hz, H-4), 5.34 (1H, dd, J.sub.3,4=10.0 Hz, J.sub.3,2=3.3 Hz, H-3), 6.18 (1H, bs, NH).

[1098] .sup.13C NMR (100.6 MHz, CDCl.sub.3): =17.8 (CH.sub.3, CH.sub.3-lactic acid), 20.61 (2CH.sub.3, 2AcO), 20.65 (CH.sub.3, AcO), 20.8 (CH.sub.3, AcO), 21.0 (CH.sub.3, AcO), 25.8 (CH.sub.2), 26.5 (CH.sub.2), 28.8 (CH.sub.2), 29.0 (CH.sub.2), 29.3 (CH.sub.2), 39.1 (CH.sub.2, C-7), 62.4 (CH.sub.2, C-6), 66.1 (CH, C-5), 68.3 (CH, CH.sub.2, C-3, C-1), 69.1 (CH, C-4), 69.6 (CH, C-2), 70.6 (CH, CH-lactic acid), 97.4 (CH, C-1), 169.4 (C, AcO), 169.7 (C, AcO), 169.9 (C, AcO), 170.0 (C, AcO), 170.2 (C, AcO), 170.6 (C, amide).

[1099] MS (CI, NH.sub.3): m/z 576 [M].sup.+

[1100] HRMS (MALDI, DHB): m/z calcd for C.sub.26H.sub.41NO.sub.13Na [M+Na ].sup.+: 598.2470, found: 598.2471.

##STR00262##

i. NaOMe, MeOH, rt, ii. Amberlite IR120 (H).

Example 10

Compound 3

[1101] According to the general procedure B, using the amide 1 (47 mg, 0.088 mmol) as starting material, the derivative 3 was obtained after lyophilization (31 mg, 0.085 mmol, 97%) as an amorphous white solid.

[1102] [].sub.D=+59.3 (c=1.19 in MeOD).

[1103] .sup.1H NMR (300 MHz, MeOD): =0.95 (3H, d, J=7.4 Hz, CH.sub.3-butyric acid), 1.30-1.70 (12H, m), 2.16 (2H, t, J=7.4 Hz, CH.sub.2-butyric acid), 3.17 (2H, t, J=6.9 Hz, C-7), 3.43 (1H, m, H-1a), 3.53 (1H, ddd, J.sub.5,4=9.3 Hz, J.sub.5,6b=5.8 Hz, J.sub.5,6a=2.4 Hz, H-5), 3.62 (1H, dd, J.sub.4,3=9.5 Hz, J.sub.4,5=9.2 Hz, H-4), 3.68-3.76 (3H, m, H-3, H-6a, 1b), 3.79 (1H, dd, J.sub.2,3=3.2 Hz, J.sub.2,1=1.7 Hz, H-2), 3.82 (1H, dd, J.sub.6b,6a=11.8 Hz, J.sub.6b,5=2.3 Hz, H-6b), 4.73 (1H, bs, H-1).

[1104] .sup.13C NMR(100.6 MHz, MeOD): =13.96 (CH.sub.3, CH.sub.3-butyric acid), 20.0 (CH.sub.3, CH.sub.3-butyric acid), 27.2 (CH.sub.2), 27.9 (CH.sub.2), 30.1 (CH.sub.2), 30.4 (CH.sub.2), 30.5 (CH.sub.2), 39.0 (CH.sub.2, CH.sub.2-butyric acid), 40.3 (CH.sub.2, C-7), 62.9 (CH, C-6), 68.5 (CH, C-1), 68.6 (CH, C-4), 72.3 (CH, C-5), 72.6 (CH, C-3), 74.6 (CH, C-2), 101.5 (CH, C-1), 176.5 (C, amide).

[1105] MS (CI, NH.sub.3): m/z 364 [M+H].sup.+

[1106] HRMS (MALDI, DHB): m/z calcd for C.sub.17H.sub.33N.sub.1O.sub.7Na [M+Na].sup.+: 386.2149, found: 386.2151

Example 11

Compound 4

[1107] According to the general procedure B, using the amide 2 (42 mg, 0.073 mmol) as starting material, the butyric derivative 4 was obtained after lyophilization (28 mg, 0.069 mmol, 94%) as an amorphous white solid.

[1108] [].sub.D=+20.7 (c=0.83 in MeOH).

[1109] .sup.1H NMR (400 MHz, MeOD): =1.29-1.41 (8H, m), 1.33 (3H, d, J=6.8 Hz, CH.sub.3-lactic acid), 1.53 (1H, m), 1.59 (1H, m), 3.21 (2H, t, J=7.1 Hz, H-7), 3.42 (1H, m, H-1a), 3.52 (1H, ddd, J.sub.5,4=9.5 Hz, J.sub.5,6b=5.7 Hz, J.sub.5,6a=2.3 Hz, H-5), 3.61 (1H, dd, J.sub.4,5=9.5 Hz, J.sub.4,3=9.5 Hz, H-4), 3.67-3.76 (3H, m, H-1b, H-3, H-6a), 3.78 (1H, dd, J.sub.2,3=3.2 Hz, J.sub.2,1=1.8 Hz, H-2), 3.82 (1H, dd, J.sub.6b,6a=11.8 Hz, J.sub.6b,5=2.3 Hz, H-6b), 4.10 (1H, q, J=6.8 Hz, CH-lactic acid), 4.73 (1H, d, J.sub.1,2=1.4 Hz, H-1).

[1110] .sup.13C NMR(100.6 MHz, MeOD): =21.3 (CH.sub.3, CH.sub.3-lactic acid), 27.2 (CH.sub.2), 27.8 (CH.sub.2), 30.1 (CH.sub.2), 30.4 (CH.sub.2), 30.5 (CH.sub.2), 39.9 (CH.sub.2, C-7), 62.9 (CH, C-6), 68.5 (CH, C-5), 68.7 (CH, C-1), 69.1 (CH), 72.3 (CH), 72.7 (CH, C-2), 74.5 (CH, CH-lactic acid), 101.5 (CH, C-1), 177.7 (C, amide).

[1111] MS (CI, NH.sub.3): m/z 424 [M+NH.sub.3].sup.+

[1112] HRMS (ESI): m/z calcd for C.sub.16H.sub.31NO.sub.8Na [M+Na].sup.+: 388,4128, found: 388,4132

##STR00263##

[1113] a) (R)-N-Boc-1-azidopropan-2-amine or 2-(azidomethy)pyridine, CuSO.sub.4, VitC Na, 1,4-dioxane-H.sub.2O, 50 C.

Example 12

Compound 5

[1114] To a solution of mannosyl alkine 3 (100 mg, 0.200 mmol) and (R)-N-Boc-1-azidopropan-2-amine (60 mg, 0.300 mmol) in a mixture 3:1 of 1,4-dioxane-H.sub.2O (4 ml) were added CuSO.sub.4 (6 mg, 0.040 mmol) and VitC Na (16 mg, 0.080 mmol) and the mixture was warmed up at 50 C. After 8 h, the mixture was concentrated and the crude was purified by silica gel column chromatography (hexanes/AcOEt: 50/50.fwdarw.10/90 as eluents) to give the triazol 5 (128 mg, 0.183 mmol, 91%) as a colorless oil.

[1115] [].sub.D=+63 (c=0.68 in CHCl.sub.3)

[1116] .sup.1H NMR (400 MHz, CDCl.sub.3): 1.08 (3H, d, J=6.8 Hz, propylamine), 1.24-1.30 (6H, m), 1.34 (9H, s, Boc), 1.52 (4H, m), 1.91 (3H, s, AcO), 1.96 (3H, s, AcO), 2.01 (3H, s, AcO), 2.07 (3H, s, AcO), 3.37 (1H, m, H-1a), 3.44 (2H, t, J=6.7 Hz, H-7), 3.59 (1H, m, H-1b), 3.90 (1H, ddd, J.sub.5,4=9.7 Hz, J.sub.5,6b=5.4 Hz, J.sub.5,6a=2.3 Hz, H-5), 3.99 (1H, m, propylamine), 4.03 (1H, dd, J.sub.6a,6b=12.3 Hz, J.sub.6a,5=2.3 Hz, H-6a), 4.19 (1H, dd, J.sub.6b,6a=12.3 Hz, J.sub.6b,5=5.4 Hz, H-6a), 4.36 (2H, m, triazol-CH.sub.2-propylamine), 4.53 (2H, bs O-CH.sub.2-triazol), 4.72 (1H, d, J.sub.1,2=1.6 Hz, H-1), 4.83 (1H, bs, NH), 5.15 (1H, dd, J.sub.2,3=3.4 Hz, J.sub.2,1=1.8 Hz, H-2), 5.19 (1H, dd, J.sub.4,3=9.9 Hz, J.sub.4,5=9.9 Hz, H-4), 5.32 (1H, dd, J.sub.3,4=10.0 Hz, J.sub.3,2=3.4 Hz, H-3), 7.50 (1H, bs, Triazol).

[1117] .sup.13C NMR (100.6 MHz, CDCl.sub.3): =20.47 (2CH.sub.3, 2AcO), 20.50 (CH.sub.3, AcO), 20.7 (CH.sub.3, AcO), 25.8 (CH.sub.3, propylamine), 28.1 (3CH.sub.3, N-Boc), 28.6-28.9 (4CH.sub.2), 29.4 (CH.sub.2), 62.5 (CH, C-6), 64.1 (CH.sub.2, O-CH.sub.2-triazol), 64.3 (CH.sub.2, triazol-CH.sub.2 propylamine), 68.2 (CH, C-5), 68.3 (CH, C-1), 68.9 (CH, C-4), 69.5 (CH, C-2), 70.5 (CH, propylamine), 70.8 (CH.sub.2, C-7), 97.3 (CH, C-1), 123.1 (CH, triazol), 145.2 (C, triazol), 154.9 (C, N-Boc), 169.5 (C, AcO), 169.7 (C, AcO), 169.9 (C, AcO), 170.4 (C, AcO).

[1118] MS (CI, NH.sub.3): m/z 702 [M+H].sup.+

[1119] HRMS (MALDI, DHB): m/z calcd for C.sub.32H.sub.52N.sub.4O.sub.13Na [M+Na].sup.+: 723.3423, found: 723.3430.

Example 13

Compound 6

[1120] To a solution of mannosyl alkine 3 (100 mg, 0.200 mmol) and 2-(azidomethy)pyridine (40 mg, 0.300 mmol) in a mixture of 3:1 of 1,4-dioxane-H.sub.2O (4 ml) were added CuSO.sub.4 (6 mg, 0.040 mmol) and VitC Na (16 mg, 0.080 mmol) and the mixture was warmed up at 50 C. After 8 h, the mixture was concentrated and the crude was purified by silica gel column chromatography (DCM.fwdarw.DCM/MeOH: 90/10 as eluents) to give the triazol 6 (120 mg, 0.191 mmol, 95%) as a colorless oil.

[1121] [].sub.D=+51.2 (c=0.91 in CHCl.sub.3)

[1122] .sup.1H NMR (300 MHz, CDCl.sub.3): 1.28-1.35 (8H, m), 1.57 (2H, m), 1.97 (3H, s, AcO), 2.02 (3H, s, AcO), 2.08 (3H, s, AcO), 2.13 (3H, s, AcO), 3.31 (1H, m, H-1a), 3.49 (2H, t, J=6.7 Hz, H-7), 3.64 (1H, m, H-1b), 3.95 (1H, ddd, J.sub.5,4=9.5 Hz, J.sub.5,6b=5.3 Hz, J.sub.5,6a=2.4 Hz, H-5), 4.08 (1H, dd, J.sub.6a,6b=12.3 Hz, J.sub.6a,5=2.3 Hz, H-6a), 4.26 (1H, dd, J.sub.6b,6a=12.3 Hz, J.sub.6b,5=5.3 Hz, H-6a), 4.59 (2H, s, O-CH.sub.2-triazol), 4.67 (1H, d, J.sub.1,2=1.7 Hz, H-1), 5.21 (1H, dd, J.sub.2,3=3.3 Hz, J.sub.2,1=1.7 Hz, H-2), 5.25 (1H, dd, J.sub.4,3=10.0 Hz, J.sub.4,5=9.8 Hz, H-4), 5.32 (1H, dd, J.sub.3,4=10.0 Hz, J.sub.3,2=3.3 Hz, H-3), 5.68 (2H, s, triazol-CH.sub.2-Py), 7.17 (1H, bd, J=7.8 Hz, Py), 7.25 (1H, ddd, J=7.6 Hz, J=4.9 Hz, J=1.1 Hz, Py), 7.67 (1H, ddd, J=7.8 Hz, J=7.8 Hz, J=1.8 Hz, Py), 7.68 (1H, bs, Triazol), 8.57 (1H, ddd, J=4.9 Hz, J=1.8 Hz, J=0.9 Hz, Py).

[1123] .sup.13C NMR (100.6 MHz, CDCl.sub.3): =20.65 (2CH.sub.3, 2AcO), 20.68 (CH.sub.3, AcO), 20.9 (CH.sub.3, AcO), 25.9-29.5 (5CH.sub.2), 55.6 (CH.sub.2, triazol-CH.sub.2-Py), 62.5 (CH, C-6), 64.3 (CH.sub.2, O-CH.sub.2-triazol), 66.2 (CH, C-4), 68.3 (CH, C-5), 68.4 (CH, C-1), 69.1 (CH, C-3), 69.7 (CH, C-2), 70.8 (CH.sub.2, C-7), 97.5 (CH.sub.2, C-1), 122.4 (CH, Py), 122.9 (CH, triazol), 123.4 (CH, Py), 137.3 (CH, Py), 145.8 (C, triazol), 149.7 (CH, Py), 154.4 (C, Py), 169.7 (C, AcO), 169.8 (C, AcO), 170.0 (C, AcO), 170.6 (C, AcO).

[1124] MS (CI, NH.sub.3): m/z 635 [M].sup.+

[1125] HRMS (MALDI, DHB): m/z calcd for C.sub.30H.sub.42N.sub.4O.sub.11Na [M+Na].sup.+: 657.2742, found: 657.2725.

##STR00264##

i. NaOMe, MeOH, rt, ii. Amberlite IR120 (H), iii. TFA-DCM, 0 C. (for N-Boc protected 5).

Example 14

Compound 7

[1126] According to the general procedure B and C, using the triazol 5 (253 mg, 0.361 mmol) as starting material, the derivative 7 was obtained after lyophilization (193 mg, 0.353 mmol, 98%), in form of trifluoroacetate salt, as an amorphous white solid.

[1127] [].sub.D=+61.3 (c=0.31 in MeOH)

[1128] .sup.1H NMR (300 MHz, MeOD): 1.31-1.66 (13H, m), 3.43 (1H, m, H-1a), 3.49-3.98 (10H, m), 4.62 (2H, s, O-CH.sub.2-triazol), 4.69 (2H, m, triazol-CH.sub.2), 4.76 (1H, bs, H-1), 8.09 (1H, bs, Triazol).

[1129] .sup.13C NMR (100.6 MHz, MeOD): =16.36 (CH.sub.3, propylamine), 27.1 (CH.sub.2), 27.2 (CH.sub.2), 30.2 (CH.sub.2), 30.4 (CH.sub.2), 30.5 (CH.sub.2), 53.4 (CH.sub.2, O-CH.sub.2-triazol), 62.5 (CH, C-6), 64.5 (CH.sub.2, triazol-CH.sub.2-propylamine), 68.4 (CH), 68.6 (CH.sub.2, C-1), 71.8 (CH), 72.2 (CH), 72.6(CH), 74.4 (CH.sub.2, C-7), 101.5 (CH, C-1), 118.3 (C, q, J.sub.C,F32 289.8 Hz, TFA), 126.2 (CH, triazol), 146.6 (C, triazol), 163.1 (C, q, J.sub.C,F=33.7 Hz, TFA).

[1130] MS (CI, NH.sub.3): m/z 433 [MTFA].sup.+

Example 15

Compound 8

[1131] According to the general procedure B, using the triazol 6 (100 mg, 0.157 mmol) as starting material, the pyridin derivative 8 was obtained after lyophilization (72 mg, 0.154 mmol, 98%) as an amorphous white solid.

[1132] [].sub.D=+36.3 (c=0.41 in MeOH)

[1133] .sup.1H NMR (300 MHz, MeOD): 1.29-1.41 (6H, m), 1.53-1.62 (4H, m), 3.40 (1H, m, H-1a), 3.50 (2H, t, J=6.6 Hz, H-7), 3.54-3.75 (6H, m), 3.81 (1H, bd, J.sub.2,3=3.8 Hz, H-2), 4.58 (2H, s, O-CH.sub.2-triazol), 4.74 (1H, bs, H-1), 4.95 (2H, s, OH), 4.97 (1H, s, OH), 5.72 (2H, s, triazol-CH.sub.2-Py), 7.33 (1H, bd, J=8.0 Hz, Py), 7.38 (1H, dd, J=7.8 Hz, J=5.3 Hz, Py), 7.84 (1H, ddd, J=7.8 Hz, J=7.8 Hz, J=1.5 Hz, Py), 8.06 (1H, bs, Triazol), 8.54 (1H, bd, J=4.5 Hz, Py).

[1134] .sup.13C NMR(100.6 MHz, MeOD): =27.1 (CH.sub.2), 27.3 (CH.sub.2), 30.2 (CH.sub.2), 30.5 (CH.sub.2), 30.6 (CH.sub.2), 56.0 (CH.sub.2, triazol-CH.sub.2-Py), 62.7 (CH, C-6), 64.6 (CH.sub.2, O-CH.sub.2-triazol), 68.5 (CH.sub.2, C-1), 68.6 (CH), 71.6 (CH.sub.2, C-7), 72.3 (CH, C-2), 72.7 (CH), 74.5 (CH), 101.6 (CH, C-1), 123.9 (CH, Py), 124.9 (CH, Py), 125.7 (CH, triazol), 139.2 (CH, Py), 146.5 (C, triazol), 150.6 (CH, Py), 155.9 (C, Py).

[1135] MS (CI, NH.sub.3): m/z 467 [M].sup.+

[1136] HRMS (MALDI, DHB): m/z calcd for C.sub.22H.sub.34N.sub.4O.sub.7Na [M+Na].sup.+: 489.2320, found: 489.2314.

##STR00265##

[1137] a) CuSO.sub.4, VitC Na, DMF-H.sub.2O, 70 C. b) i. NaOMe, MeOH, rt, ii. Amberlite IR120 (H).

Example 16

Compound 10

[1138] Alkynyl-saccharide 4 (87 mol) and mono-7-azido-7-deoxy-gamma-cyclodextrin (43 mol) were dissolved in a DMF/H.sub.2O mixture (2/0.5 mL). Copper sulfate (43 mol) and sodium ascorbate (86 mol) were added and the mixture was stirred at 70 C. for 30 minutes under W irradiation. Ethylenediamine tetraacetic acid trisodium salt (127 mol) was added and the mixture was stirred for 10 minutes at rt. The mixture was evaporated under reduced pressure and the residue purified by preparative HPLC leading to compound 10 as a white powder after lyophilisation.

##STR00266##

[1139] a) Pyridinmethyl propargyl ether, CuSO.sub.4, VitC Na, 1,4-dioxane-H.sub.2O, 50 C. b) NaOMe, MeOH.

Example 18

Compound 11

[1140] To a solution of mannosyl azide 6 (100 mg, 0.205 mmol) and pyridinmethyl propargyl ether (36 mg, 0.246 mmol) in a mixture of 3:1 of 1,4-dioxane-H.sub.2O (4.1 ml) were added CuSO.sub.4 (7 mg, 0.041 mmol) and VitC Na (16 mg, 0.082 mmol) and the mixture was warmed up at 65 C. After 8 h, the mixture was concentrated and the crude was purified by silica gel column chromatography (AcOEt.fwdarw.AcOEt/MeOH: 90/10 as eluents) to give the triazol 11 (128 mg, 0.202 mmol, 98%) as a colorless oil.

[1141] [].sub.D=+37.9 (c=0.83 in CHCl.sub.3)

[1142] .sup.1H NMR (300 MHz, CDCl.sub.3): 1.29-1.41 (6H, m), 1.58 (2H, m), 1.91 (2H, m), 1.99 (3H, s, AcO), 2.04 (3H, s, AcO), 2.09 (3H, s, AcO), 2.15 (3H, s, AcO), 3.42 (1H, m, H-1a), 3.67 (1H, m, H-1b), 3.96 (1H, ddd, J.sub.5,4=9.3 Hz, J.sub.5,6b=5.3 Hz, J.sub.5,6a=2.4 Hz, H-5), 4.10 (1H, dd, J.sub.6a,6b=12.3 Hz, J.sub.6a,5=2.3 Hz, H-6a), 4.28 (1H, dd, J.sub.6b,6a=12.3 Hz, J.sub.6b,5=5.3 Hz, H-6a), 4.35 (2H, t, J=7.3 Hz, H-7), 4.72 (2H, s, O-CH.sub.2-triazol), 4.77 (2H, s, O-CH.sub.2-Py), 4.79 (1H, d, J.sub.1,2=1.7 Hz, H-1), 5.22 (1H, dd, J.sub.2,3=3.3 Hz, J.sub.2,1=1.7 Hz, H-2), 5.27 (1H, dd, J.sub.4,3=10.1 Hz, J.sub.4,5=9.7 Hz, H-4), 5.34 (1H, dd, J.sub.3,4=10.1 Hz, J.sub.3,2=3.3 Hz, H-3), 7.19 (1H, dd, J=7.4 Hz, J=5.2 Hz, Py), 7.55 (1H, bd, J=7.8 Hz, Py), 7.59 (1H, bs, Triazol), 7.69 (1H, ddd, J=7.8 Hz, J=7.8 Hz, J=1.8 Hz, Py), 8.56 (1H, bd, J=4.8 Hz, Py).

[1143] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =20.73-20.88 (4CH.sub.3, 4AcO), 25.9 (CH.sub.2), 26.7 (CH.sub.2), 28.7 (CH.sub.2), 29.1 (CH.sub.2), 30.2 (CH.sub.2), 50.3 (CH.sub.2, C-7), 62.5 (CH, C-6), 64.4 (CH.sub.2, O-CH.sub.2-triazol), 66.2 (CH, C-4), 68.35 (CH, C-1), 68.40 (CH, C-5), 69.1 (CH, C-3), 69.7 (CH, C-2), 73.3 (CH.sub.2, O-CH.sub.2-Py), 97.5 (CH.sub.2, C-1), 121.7 (CH, Py), 122.4 (CH, Py),122.5 (CH, triazol), 136.7 (CH, Py), 144.8 (C, triazol), 149.2 (CH, Py), 157.9 (C, Py), 169.7 (C, AcO), 169.9 (C, AcO), 170.1 (C, AcO), 170.6 (C, AcO).

[1144] MS (MALDI): m/z 657 [M+Na].sup.+

[1145] HRMS (MALDI, DHB): m/z calcd for C.sub.30H.sub.42N.sub.4O.sub.11[M].sup.+: 635.2923, found: 635.2944.

Example 19

Compound 12

[1146] According to the general procedure B, using the triazol 11 (110 mg, 0.173 mmol) as starting material, the pyridin derivative 12 was obtained after lyophilization (7 mg, 0.154 mmol, 98%) as an amorphous white solid.

[1147] [].sub.D=+51.2 (c=0.49 in MeOH)

[1148] .sup.1H NMR (300 MHz, MeOD): 1.38 (6H, m), 1.58 (2H, m), 1.93 (2H, m), 1.99 (3H, s, AcO), 3.41 (1H, m, H-1a), 3.48-3.85 (7H, m), 4.43 (2H, t, J=7.1 Hz, H-7), 4.69 (2H, s, O-CH.sub.2-triazol), 4.77 (3H, bs, O-CH.sub.2-Py, H-1), 7.37 (1H, dd, J=7.3 Hz, J=5.1 Hz, Py), 7.56 (1H, bd, J=7.9 Hz, Py), 7.86 (1H, ddd, J=7.8 Hz, J=7.8 Hz, J=1.8 Hz, Py), 8.06 (1H, s, Triazol), 8.50 (1H, bd, J=4.8 Hz, Py).

[1149] .sup.13C NMR(100.6 MHz, MeOD): =27.1 (CH.sub.2), 27.3 (CH.sub.2), 29.8 (CH.sub.2), 30.4 (CH.sub.2), 31.2 (CH.sub.2), 51.3 (CH.sub.2, C-7), 62.8 (CH, C-6), 64.7 (CH.sub.2, O-CH.sub.2-triazol), 68.4 (CH, C-1), 68.6 (CH), 72.3 (CH), 72.7 (CH), 73.4 (CH.sub.2, O-CH.sub.2-Py), 74.6 (CH), 101.5 (CH, C-1), 123.4 (CH, Py), 124.2 (CH, Py),125.2 (CH, Py), 138.9 (CH, triazol), 145.6 (C, triazol), 149.6 (CH, Py), 159.0 (C, Py).

[1150] MS (CI, NH.sub.3): m/z 467 [M+H].sup.+

C. Synthesis of Mannosyl-S-heptylamides

[1151] ##STR00267##

[1152] a) 7-bromo-l-heptanol, Et.sub.2NH, DMF, rt. b) CCl.sub.4, Ph.sub.3P, DCM, 0 C..fwdarw.rt. c) NaN.sub.3, DMF, 70 C. d) Carboxylic acid, HOBt, DIC, PH.sub.3P, THF, 0 C..fwdarw.rt

Example 20

Compound 14

[1153] To a solution of acetylated 1-thiosugar 13 (1.28 g, 3.15 mmol) and 7-bromo-1-heptanol (738 mg, 3.78 mmol, 1.2 equiv) in dry DMF (150 mL) at room temperature under a nitrogen atmosphere, was added diethylamine (6.51 mL, 63.06 mmol, 20 equiv). After stirring for 8 hours, diethylamine and dimethylformamide were removed in vacuo. The crude product was purified by silica gel column chromatography (Hexanes/EtOAc, 50:50) to give the product 14 (1.42 g, 2.97 mmol, 94%) as an amorphous white solid.

[1154] [].sub.D=+83.2 (c=1.13 in CHCl.sub.3)

[1155] .sup.1NMR (300 MHz, CDCl.sub.3): 1.34 (6H, m), 1.62 (2H, m), 1.85 (2H, m), 1.99 (3H, s, AcO), 2.05 (3H, s, AcO), 2.09 (3H, s, AcO), 2.16 (3H, s, AcO), 2.61 (2H, m, H-1), 3.40 (2H, t, J=6.8 Hz, H-7), 4.08 (1H, dd, J.sub.6a,6b=11.9 Hz, J.sub.6a,5=1.9 Hz, H-6a), 4.32 (1H, dd, J.sub.6b,6a=11.9 Hz, J.sub.6b,5=5.3 Hz, H-6a), 4.38 (1H, m, H-5), 5.25 (1H, d, J.sub.1,2=1.4 Hz, H-1), 5.24-5.35 (2H, m, H-3, H-4), 5.35 (1H, dd, J.sub.2,3=2.8 Hz, J.sub.2,1=1.4 Hz, H-2).

[1156] .sup.13C NMR (100.6 MHz, CDCl.sub.3): =3.9 (CH3), 58.7 (CH3), 59.2 (CH3), 60.5 (CH3), 60.8 (CH3), 66.2 (CH), 71.3 (CH2), 72.9 (CH), 73.4 (C), 79.4 (CH), 81.0 (CH), 84.5 (CH).

[1157] MS (CI, NH.sub.3): m/z: [M+NH.sub.3].sup.+496

[1158] HRMS (MALDI, DHB): m/z calcd for C.sub.21H.sub.34O.sub.10SNa [M+Na].sup.+: 501.1765, found: 501.1785

Example 21

Compound 15

[1159] A solution of 14 (1.35 g, 2.82 mmol) and carbon tetrabromide (1.03 g, 3.10 mmol) in dry DCM (15 mL), cooled to 0 C. was added Ph.sub.3P (812 mg, 3.10 mmol) in portions over 30 min with vigorous stirring. Upon addition of the phosphine, the colorless solution turned a pale brown color and was stirred for an additional 2 h at room temperature. The mixture was concentrated and the crude was purified by silica gel column chromatography (Hexanes/EtOAc, 80:20) to give the product 15 (1.41 g, 2.61 mmol, 93%) as an amorphous white solid.

[1160] [].sub.D=+83.8 (c=0.79 in CHCl.sub.3)

[1161] .sup.1H NMR (300 MHz, CDCl.sub.3): 1.34 (6H, m), 1.62 (2H, m), 1.85 (2H, m), 1.99 (3H, s, AcO), 2.05 (3H, s, AcO), 2.09 (3H, s, AcO), 2.16 (3H, s, AcO), 2.61 (2H, m, H-1), 3.40 (2H, t, J=6.8 Hz, H-7), 4.08 (1H, dd, J.sub.6a,6b=11.9 Hz, J.sub.6a,5=1.9 Hz, H-6a), 4.32 (1H, dd, J.sub.6b,6a=11.9 Hz, J.sub.6b,5=5.3 Hz, H-6a), 4.38 (1H, m, H-5), 5.25 (1H, d, J.sub.1,2=1.4 Hz, H-1), 5.24-5.35 (2H, m, H-3, H-4), 5.35 (1H, dd, J.sub.2,3=2.8 Hz, J.sub.2,1=1.4 Hz, H-2).

[1162] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =20.5 (CH.sub.3, AcO), 20.6 (CH.sub.3, AcO), 20.63 (CH.sub.3, AcO), 20.8 (CH.sub.3, AcO), 27.9 (CH.sub.2), 28.1 (CH.sub.2), 28.4 (CH.sub.2), 29.1 (CH.sub.2), 31.1 (CH.sub.2, C-1), 32.5 (CH.sub.2), 33.7 (CH.sub.2, C-7), 62.3 (CH, C-6), 66.2 (CH, C-3 or C-4), 68.8 (CH, C-5), 69.3 (CH, C-3 or C-4), 71.1 (CH, C-2), 82.4 (CH, C-1), 169.6 (C, AcO), 169.7 (C, AcO), 169.9 (C, AcO), 170.5 (C, AcO).

[1163] MS (CI, NH.sub.3): m/z: [M+NH.sub.3].sup.+560

[1164] HRMS (MALDI, DHB): m/z calcd for C.sub.21H.sub.33BrO.sub.9SNa [M+Na].sup.+: 563.0921, found: 563.0932

Example 22

Compound 16

[1165] A solution of 15 (560 mg, 1.037 mmol) in DMF (10 mL) was added NaN.sub.3 (135 mg, 2.074 mmol) and the resulting mixture was stirred at 70 C. overnight. The mixture was diluted with Et.sub.2O and washed with H.sub.2O and brine. The crude was purified by silica gel column chromatography (Hexanes/EtOAc, 70:30) to give the azide 16 (498 mg, 0.990 mmol, 96%) as a colorless oil.

[1166] [].sub.D=+73.3 (c=0.67 in CHCl.sub.3)

[1167] .sup.1H NMR (300 MHz, CDCl.sub.3): 1.25-1.65 (10H, m), 1.99 (3H, s, AcO), 2.05 (3H, s, AcO), 2.09 (3H, s, AcO), 2.16 (3H, s, AcO), 2.60 (2H, m, H-1), 3.26 (2H, t, J=6.9 Hz, H-7), 4.08 (1H, dd, J.sub.6a,6b=11.9 Hz, J.sub.6a,5=2.0 Hz, H-6a), 4.32 (1H, dd, J.sub.6b,6a=11.9 Hz, J.sub.6b,5=5.2 Hz, H-6a), 4.38 (1H, m, H-5), 5.23-5.35 (3H, m, H-1, H-3, H-4), 5.33 (1H, dd, J.sub.2,3=2.9 Hz, J.sub.2,1=1.5 Hz, H-2).

[1168] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =20.6 (CH.sub.3, AcO), 20.7 (CH.sub.3, AcO), 20.73 (CH.sub.3, AcO), 20.9 (CH.sub.3, AcO), 26.5 (CH.sub.2), 28.5 (CH.sub.2), 28.6 (CH.sub.2), 28.7 (CH.sub.2), 29.2 (CH.sub.2), 31.2 (CH.sub.2, C-1), 51.3 (CH.sub.2, C-7), 62.4 (CH, C-6), 66.2 (CH, C-3 or C-4), 68.9 (CH, C-5), 69.4 (CH, C-3 or C-4), 71.1 (CH, C-2), 82.4 (CH, C-1), 169.7 (C, AcO), 169.72 (C, AcO), 169.9 (C, AcO), 170.5 (C, AcO).

[1169] MS (CI, NH.sub.3): m/z: [M+NH.sub.3].sup.+521

[1170] HRMS (MALDI, DHB): m/z calcd for C.sub.21H.sub.33N.sub.3O.sub.9SNa [M+Na].sup.+: 526.1830, found: 526.1836

Example 23

Compound 17

[1171] According to the general procedure A, mannosyl azide 16 (50 mg, 0.096 mmol), acetic acid (11 mg, 0.173 mmol, 1.8 equiv.), HOBt (23 mg, 0.173 mmol, 1.8 equiv.), DIC (27 L, 0.173 mmol, 1.8 equiv.) and Ph.sub.3P (45 mg, 0.173 mmol, 1.8 equiv.) were allowed to react in THF (2.4 mL). The crude product was purified by silica gel column chromatography (EtOAc/petroleum ether, 70:30.fwdarw.EtOAc as eluents) to give the amide 17 (39 mg, 0.075 mmol, 78%) as an oil.

[1172] [].sub.D=+69.5 (c=0.81 in CHCl.sub.3)

[1173] .sup.1H NMR (300 MHz, CDCl.sub.3): 1.27-1.65 (10H, m), 1.96 (3H, s, AcO), 1.98 (3H, s, AcO), 2.04 (3H, s, AcO), 2.09 (3H, s, AcO), 2.15 (3H, s, AcO), 2.59 (2H, m, H-1), 3.21 (2H, q, J=6.8 Hz, H-7), 4.08 (1H, dd, J.sub.6a,6b=12.0 Hz, J.sub.6a,5=2.1 Hz, H-6a), 4.30 (1H, dd, J.sub.6b,6a=12.0 Hz, J.sub.6b,5=5.1 Hz, H-6a), 4.36 (1H, m, H-5), 5.22-5.34 (3H, m, H-1, H-3, H-4), 5.32 (1H, dd, J.sub.2,3=3.0 Hz, J.sub.2,1=1.6 Hz, H-2), 5.57 (1H, bs, NH).

[1174] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =20.6 (CH.sub.3, AcO), 20.70 (CH.sub.3, AcO), 20.75 (CH.sub.3, AcO), 20.9 (CH.sub.3, AcO), 23.3 (CH.sub.3, acetamide), 26.7 (CH.sub.2), 28.5 (CH.sub.2), 28.7 (CH.sub.2), 29.1 (CH.sub.2), 29.5 (CH.sub.2), 31.2 (CH.sub.2, C-1), 39.5 (CH.sub.2, C-7), 62.4 (CH, C-6), 66.2 (CH, C-3 or C-4), 68.9 (CH, C-5), 69.4 (CH, C-3 or C-4), 71.2 (CH, C-2), 82.4 (CH, C-1), 169.7 (C, acetamide), 169.8 (C, AcO), 169.96 (C, AcO), 169.99 (C, AcO), 170.6 (C, AcO).

[1175] MS (CI, NH.sub.3): m/z: [M].sup.+ 520

[1176] HRMS (MALDI, DHB): m/z calcd for C.sub.27H.sub.38N.sub.2O.sub.10SNa [M+Na].sup.+: 542.2036, found: 542.2028

Example 24

Compound 18

[1177] According to the general procedure A, mannosyl azide 16 (150 mg, 0.289 mmol), N-Boc-L-alanine (98 mg, 0.520 mmol, 1.8 equiv.), HOBt (70 mg, 0.520 mmol, 1.8 equiv.), DIC (80 L, 0.520 mmol, 1.8 equiv.) and Ph.sub.3P (136 mg, 0.520 mmol, 1.8 equiv.) were allowed to react in THF (27.3 mL). The crude product was purified by silica gel column chromatography (EtOAc/petroleum ether, 50:50.fwdarw.EtOAc as eluents) to give the amide 18 (97 mg, 0.149 mmol, 52%) as an oil.

[1178] [].sub.D=+39.9 (c=1.27 in CHCl.sub.3)

[1179] .sup.1H NMR (300 MHz, CDCl.sub.3): =1.11-1.63 (10H, m),1.34 (3H, d, J=7.1 Hz, CH.sub.3-alanine), 1.42 (9H, s, N-Boc), 1.98 (3H, s, AcO), 2.04 (3H, s, AcO), 2.07 (3H, s, AcO), 2.15 (3H, s, AcO), 2.53 (2H, m, H-1), 2.99 (2H, m, H-7), 4.04-4.38 (4H, m, H-5, H-6a, H-6b, CH-alanine), 4.21-4.29 (3H, m, H-1, H-3, H-4), 5.32 (1H, dd, J.sub.2,3=3.0 Hz, J.sub.2=1.7 Hz, H-2), 5.36 (1H, bs, NH), 5.66 (1H, bs, NH).

[1180] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =20.5 (CH.sub.3, AcO), 20.60 (CH.sub.3, AcO), 20.63 (CH.sub.3, AcO), 20.8 (CH.sub.3, AcO), 26.5 (CH.sub.3, alanine), 28.2 (3 X CH.sub.3, N-Boc), 28.5 (CH.sub.2), 28.6 (CH.sub.2), 29.1 (CH.sub.2), 29.3 (CH.sub.2), 30.2 (C, N-Boc), 31.1 (CH.sub.2), 39.3 (CH.sub.2, C-7), 41.8 (CH.sub.2, C-1), 62.3 (CH, C-6), 68.8 (CH), 69.4 (CH), 71.1 (CH, C-2), 82.4 (CH, C-1), 157.1 (C, amide), 169.6 (C, AcO), 169.7 (C, AcO), 169.9 (C, AcO), 170.5 (C, AcO), 172.5 (C, N-Boc).

[1181] MS (CI, NH.sub.3): m/z: [M].sup.+649

[1182] HRMS (MALDI, DHB): m/z calcd for C.sub.29H.sub.48N.sub.2O.sub.12SNa [M+Na].sup.+: 671.2820, found: 671.2803

Example 25

Compound 19

[1183] According to the general procedure A, mannosyl azide 16 (100 mg, 0.193 mmol), picolinic acid (43 mg, 0.347 mmol, 1.8 equiv.), HOBt (47 mg, 0.347 mmol, 1.8 equiv.), DIC (54 L, 0.347 mmol, 1.8 equiv.) and Ph.sub.3P (91 mg, 0.347 mmol, 1.8 equiv.) were allowed to react in DMF (5 mL). The crude product was purified by silica gel column chromatography (DCM.fwdarw.DCM/MeOH, 90:10 as eluents) to give the amide 19 (83 mg, 0.142 mmol, 74%) as an oil.

[1184] [].sub.D=+55.7 (c=1.01 in CHCl.sub.3)

[1185] .sup.1H NMR (300 MHz, CDCl.sub.3): 1.36-1.68 (10H, m), 1.98 (3H, s, AcO), 2.04 (3H, s, AcO), 2.09 (3H, s, AcO), 2.16 (3H, s, AcO), 2.60 (2H, m, H-1), 3.46 (2H, q, J=6.8 Hz, H-7), 4.08 (1H, dd, J.sub.6a,6b=11.9 Hz, J.sub.6a,5=1.9 Hz, H-6a), 4.31 (1H, dd, J.sub.6b,6a=11.9 Hz, J.sub.6b,5=5.2 Hz, H-6a), 4.37 (1H, m, H-5), 5.23-5.30 (3H, m, H-1, H-3, H-4), 5.33 (1H, dd, J.sub.2,3=2.8 Hz, J.sub.2,1=1.6 Hz, H-2), 7.41 (1H, ddd, J=7.7 Hz, J=4.9 Hz, J=1.3 Hz, picolinic), 7.84 (1H, ddd, J=7.6 Hz, J=7.6 Hz, J=1.7 Hz, picolinic), 8.05 (1H, bs, NH), 8.19 (1H, bd, J=7.8 Hz, picolinic), 8.54 (1H, ddd, J=4.7 Hz, J=1.7 Hz, J=0.9 Hz, picolinic).

[1186] .sup.13C NMR (100.6 MHz, CDCl.sub.3): =20.53 (CH.sub.3, AcO), 20.60 (CH.sub.3, AcO), 20.63 (CH.sub.3, AcO), 20.8 (CH.sub.3, AcO), 26.7 (CH.sub.2), 28.5 (CH.sub.2), 28.7 (CH.sub.2), 29.2 (CH.sub.2), 29.5 (CH.sub.2), 31.1 (CH.sub.2, C-1), 39.2 (CH.sub.2, C-7), 62.3 (CH, C-6), 66.2 (CH, C-3 or C-4), 68.8 (CH, C-5), 69.4 (CH, C-3 or C-4), 71.1 (CH, C-2), 82.4 (CH, C-1), 122.1 (CH, picolinic acid), 126.0 (CH, picolinic acid), 137.2 (CH, picolinic acid), 147.9 (CH, picolinic acid), 149.9 (C, picolinic acid), 164.1 (C, amide), 169.6 (C, AcO), 169.7 (C, AcO), 169.9 (C, AcO), 170.5 (C, AcO).

[1187] MS (CI, NH.sub.3): m/z: [M+NH.sub.3].sup.+ 583

[1188] HRMS (MALDI, DHB): m/z calcd for C.sub.27H.sub.38N.sub.2O.sub.10SNa [M+Na].sup.+: 605.2139, found: 605.2129

##STR00268##

i. NaOMe, MeOH, rt, ii. Amberlite IR120 (H), iii. TFA-DCM, 0 C. vi. HCl ac. (for N-Boc protected compound 18).

Example 26

Compound 20

[1189] According to the general procedure B, using the amide 17 (20 mg, 0.038 mmol) as starting material, the derivative 20 was obtained after lyophilization (17 mg, 0.037 mmol, 98%) as an amorphous white solid.

[1190] [a].sub.D=+53.9 (c=0.69 in MeOD).

[1191] .sup.1H NMR (300 MHz, MeOD): =1.29-1.71 (10H, m), 1.92 (3H, s, acetamide), 2.64 (2H, m, H-7), 3.15 (2H, t, J=6.9 Hz, H-1), 3.64-3.92 (6H, m), 5.21 (1H, d, J.sub.1 2=1.3 Hz, H-1).

[1192] .sup.13C NMR (100.6 MHz, MeOD): =22.5 (CH.sub.3, acetamide), 27.8 (CH.sub.2), 29.7 (CH.sub.2), 29.9 (CH.sub.2), 30.3 (CH.sub.2), 30.6 (CH.sub.2), 31.8 (CH.sub.2, C-1), 40.5 (CH.sub.2, C-7), 62.7 (CH, C-6), 68.9 (CH), 73.1 (CH), 73.8 (CH, C-5), 74.9 (CH, C-2), 86.4 (CH, C-1), 173.2 (C, acetamide).

[1193] MS (CI, NH.sub.3): m/z 352 [M+H].sup.+

[1194] HRMS (ESI): m/z calcd for C.sub.15H.sub.29O.sub.6NSNa [M+Na].sup.+: 374.1613, found: 374.1615

Example 27

Compound 21

[1195] According to the general procedure B and C, using the amide 18 (51 mg, 0.078 mmol) as starting material, the alanine derivative 21 was obtained after lyophilization (31 mg, 0.074 mmol, 95%), in form of ammonium chloride salt, as an amorphous white solid.

[1196] [a].sub.D=+61.3 (c=0.61 in D.sub.2O).

[1197] .sup.1H NMR (300 MHz, D.sub.2O): =1.12-1.48 (10H, m), 1.36 (3H, d, J=7.1 Hz, CH.sub.3-alanine), 2.63 (2H, m, H-7), 3.42 (2H, m, H-1), 3.31-3.87 (7H, m), 4.64 (1H, bs, H-1).

[1198] .sup.13C NMR(100.6 MHz, D.sub.2O): =16.6 (CH.sub.3, alanine), 24.9 (CH.sub.2), 25.2 (CH.sub.2), 25.9 (CH.sub.2), 28.0 (CH.sub.2), 31.2 (CH.sub.2), 61.8 (CH, C-1), 39.5 (CH.sub.2, C-7), 60.9 (CH, C-6), 66.8 (CH, alanine), 67.9 (CH), 70.1 (CH), 70.7 (CH), 72.7 (CH, C-2), 85.9 (CH, C-1), 170.5 (C, amide).

[1199] MS (CI, NH.sub.3): m/z 417 [M].sup.+

[1200] HRMS (ESI): m/z calcd

Example 28

Compound 22

[1201] According to the general procedure B, using the amide 19 (32 mg, 0.055 mmol) as starting material, the derivative 22 was obtained after lyophilization (22 mg, 0.053 mmol, 96%) as an amorphous white solid.

[1202] [].sub.D=+53.9 (c=0.69 in MeOD).

[1203] .sup.1H NMR (300 MHz, MeOD): =1.28-1.69 (10H, m), 2.63 (2H, m, H-7), 3.42 (2H, m, H-1), 3.64-3.93 (6H, m), 5.21 (1H, d, J.sub.1,2=1.0 Hz, H-1), 7.41 (1H, ddd, J=7.6 Hz, J=4.8 Hz, J=1.3 Hz, picolinic), 7.53 (1H, ddd, J=7.6 Hz, J=4.8 Hz, J=1.3 Hz, picolinic), 7.95 (1H, ddd, J=7.7 Hz, J=7.7 Hz, J=1.7 Hz, picolinic), 8.09 (1H, ddd, J=7.8 Hz, J=1.1 Hz, J=1.1 Hz, picolinic), 8.62 (1H, ddd, J=4.7 Hz, J=1.7 Hz, J=1.1 Hz, picolinic).

[1204] .sup.13C NMR(100.6 MHz, MeOD): =27.9 (CH.sub.2), 29.7 (CH.sub.2), 29.9 (CH.sub.2), 30.5 (CH.sub.2), 30.6 (CH.sub.2), 31.8 (CH.sub.2, C-1), 40.4 (CH.sub.2, C-7), 62.7 (CH, C-6), 68.9 (CH), 73.2 (CH), 73.8 (CH, C-5), 74.9 (CH, C-2), 86.4 (CH, C-1), 123.0 (CH, picolinic), 127.6 (CH, picolinic), 138.8 (CH, picolinic), 149.8 (CH, picolinic), 151.1 (C, picolinic), 166.6 (C, amide).

[1205] MS (CI, NH.sub.3): m/z 415 [M+H].sup.+

[1206] HRMS (ESI): m/z calcd for C.sub.19H.sub.30N.sub.2O.sub.6SNa [M+Na].sup.+: 437.1722, found: 437.1735

##STR00269##

[1207] a) CBr.sub.4, Ph.sub.3P, DCM, 0 C..fwdarw.rt. b) 1-bromo-7-propargyloxyheptane (13), Et.sub.2NH, DMF, rt. c) CuSO.sub.4, VitC Na, DMF-H.sub.2O, 70 C. d) CuSO.sub.4, VitC Na, DMF-H.sub.2O, 70 C.

Example 29

Compound 13

[1208] A solution of 7-O-propargylheptanediol (500 mg, 2.941 mmol) and carbon tetrabromide (1.07 g, 3.235 mmol) in dry DCM (15 mL), cooled to 0 C. was added Ph.sub.3P (848 mg, 3.235 mmol) in portions over 30 min with vigorous stirring. Upon addition of the phosphine, the colorless solution turned a pale brown color and was stirred for an additional 3 h at room temperature. The mixture was concentrated and the crude was purified by silica gel column chromatography (Hexanes/EtOAc, 70:30) to give the product 13 (625 mg, 2.682 mmol, 91%) as a colorless oil.

[1209] .sup.1H NMR (300 MHz, CDCl.sub.3): =1.30-1.48 (6H, m), 1.59 (2H, m), 1.86 (2H, m), 2.41 (1H, t, J=2.4 Hz, CH-propargyl), 3.40 (2H, t, J=6.8 Hz), 3.51 (2H, t, J=6.5 Hz), 4.13 (2H, d, J=2.4 Hz, CH.sub.2-propargyl).

[1210] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =25.9 (CH.sub.2), 28.1 (CH.sub.2), 28.5 (CH.sub.2), 29.4 (CH.sub.2), 32.7 (CH.sub.2), 33.9 (CH.sub.2), 58.0 (CH.sub.2, CH.sub.2-propargyl), 70.1 (CH.sub.2), 74.1 (CH, CH-propargyl), 77.2 (C, C-propargyl).

Example 30

Compound 14

[1211] To a solution of acetylated 1-thiosugar 13 (944 mg, 2.325 mmol) and 1-bromo-7-propargyloxyheptane 13 (650 mg, 2.789 mmol, 1.2 equiv) in DMF (93 mL) at rt under a nitrogen atmosphere, was added diethylamine (4.8 mL, 46.500 mmol, 20 equiv). After stirring for 8 hours, diethylamine and DMF were removed in vacuo. The crude product was purified by silica gel column chromatography (Hexanes/EtOAc, 80:20) to give the product 14 (1026 mg, 1.984 mmol, 85%) as a colorless oil.

[1212] [].sub.D=+47.2 (c=0.28 in CHCl.sub.3)

[1213] .sup.1H NMR (300 MHz, CDCl.sub.3): 1.18-1.36 (8H, m), 1.53 (2H, m), 1.99 (3H, s, AcO), 2.05 (3H, s, AcO), 2.10 (3H, s, AcO), 2.16 (3H, s, AcO), 2.42 (1H, t, J=2.4 Hz, propargyl), 2.60 (2H, m, H-1), 3.50 (2H, t, J=3.5 Hz, H-7), 4.08 (1H, dd, J.sub.6a,6b=11.9 Hz, J.sub.6a,5=2.0 Hz, H-6a), 4.13 (2H, d, J=2.4 Hz, propargyl), 4.32 (1H, dd, J.sub.6b,6a=11.9 Hz, J.sub.6b,5=5.3 Hz, H-6b), 4.38 (1H, m, H-5), 5.25 (1H, d, J.sub.1,2=1.4 Hz, H-1), 5.27-5.34 (3H, m, H-2, H-3, H-4).

[1214] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =20.4 (CH.sub.3, AcO), 20.47 (CH.sub.3, AcO), 20.50 (CH.sub.3, AcO), 20.7 (CH.sub.3, AcO), 25.74 (CH.sub.2), 28.5 (CH.sub.2), 28.6 (CH.sub.2), 29.1 (CH.sub.2), 29.2 (CH2), 31.1 (C-1, CH.sub.2), 31.5 (CH.sub.2), 57.8 (CH.sub.2, CH.sub.2-propargyl), 62.2 (CH.sub.2, C-6), 66.1 (CH, C-3 or C-4), 68.7 (CH, C-5), 69.2 (CH, C-3 or C-4), 69.8 (CH.sub.2, C-7), 70.9 (CH, C-2), 73.9 (CH, CH-propargyl), 79.8 (C, C-propargyl), 82.3 (CH, C-1), 169.5 (C, AcO), 169.5 (C, AcO), 169.7 (C, AcO), 170.3 (C, AcO).

[1215] MS (CI, NH.sub.3): m/z: [M+NH.sub.3].sup.+534

[1216] HRMS (MALDI, DHB): m/z calcd for C.sub.24H.sub.36O.sub.10S [M+Na].sup.+: 539.1921, found: 539.1945.

Example 31

Compound 15

[1217] To a solution of 6-azido-2,3,6-O-acetyl--Cyclodextrin (150 mg, 0.075 mmol) and alkyne 14 (47 mg, 0.090 mmol) in a mixture DMF-H.sub.2O (3:1, 3.8 ml) were added CuSO.sub.4 (2 mg, 0.015 mmol) and VitC Na (6 mg, 0.030 mmol) and the mixture was warmed up at 60 C. After 12 h, the mixture was diluted with water, extracted with AcOEt, dried, concentrated and the crude was purified by silica gel column chromatography (AcOEt.fwdarw.AcOEt/MeOH: 95/5 as eluents) to give the monovalent derivative 15 (128 mg, 0.051 mmol, 68%) as a colorless solid.

[1218] [].sub.D=+101.7 (c=0.21 in CHCl.sub.3)

[1219] .sup.1H NMR (400 MHz, CDCl.sub.3) =1.22-1.44 (6H, m), 1.60 (4H, m), 1.97-2.15 (78H, m, 26AcO), 2.59 (2H, m, H-1), 3.50 (2H, t, J=6.7 Hz, H-7), 3.54-3.78 (8H, m), 4.03-4.70 (18H, m), 4.72-4.86 (6H, m, 6H-2 CD), 4.94 (1H, dd, J=8.4 Hz, J=3.6 Hz, H-2 CD), 5.00-5.13 (6H, m, 6H-1 CD), 5.15-5.38 (12H, m), 5.64 (1H, d, J=3.9 Hz, H-1 CD), 7.59 (1H, s, triazol).

[1220] .sup.13C NMR (125 MHz, CDCl.sub.3): =20.3-20.7 (24CH.sub.3, AcO), 25.8 (CH.sub.2), 28.8 (CH.sub.2), 29.9 (CH.sub.2), 29.3 (CH.sub.2), 29.4 (CH.sub.2), 31.2 (CH.sub.2, C-1 thioglycoside), 50.4 (CH.sub.2, C-6 CD), 59.5-82.9 (9CH.sub.2, 32CH), 82.6 (CH, C-1 thioglycoside), 96.0-96.6 (7CH, C-1 CD), 125.5 (CH, triazol), 145.8 (C, triazol), 169.1-171.2 (24C, AcO).

[1221] HRMS (ESI): m/z calcd for C.sub.106H.sub.145N.sub.3O.sub.64SNa.sub.2 [M+2Na].sup.2+: 1280.8844, found: 1280.8864.

##STR00270##

i. NaOMe, MeOH, rt, ii. Amberlite IR120 (H)

Example 32

Compound 16

[1222] According to the general procedure B, using the derivative 15 (55 mg, 0.029 mmol) as starting material, the derivative 16 was obtained after lyophilization (39 mg, 0.026 mmol, 89%) as an amorphous white solid.

[1223] [].sub.D=+161 (c=1.12, MeOH)

[1224] .sup.1H NMR (400 MHz, D.sub.2O) =1.14-1.74 (10H, m), 2.77 (2H, m, H-1), 3.15 (1H, bd, J.sub.6a,6b=11.8 Hz, H-6a thioglycoside), 3.38-4.18 (51H, m), 5.11 (1H, d, J=3.5 Hz, H-1 CD), 5.17 (5H, m, H-1 CD), 5.31 (1H, d, J=3.5 Hz, H-1 CD), 5.42 (1H, d, J=1.1 Hz, H-1 thioglycoside), 8.04 (1H, s, triazol).

[1225] .sup.13C NMR (125 MHz, D.sub.2O): =25.3 (CH.sub.2), 27.4 (CH.sub.2), 27.5 (CH.sub.2), 28.2 (CH.sub.2), 28.4 (CH.sub.2), 30.5 (CH.sub.2, C-1 thioglycoside), 51.5 (CH.sub.2, C-6 CD), 58.9-83.2 (9CH.sub.2, 32CH), 85.2 (CH, C-1 thioglycoside), 101.9-102.3 (7CH, C-1 CD), 123.8 (CH, triazol), 146.2 (C, triazol). HRMS (MALDI, DHB): m/z calcd for C.sub.58H.sub.97N.sub.3O.sub.40SNa [M+Na].sup.+: 1530.5261, found: 1530.5252.

[1226] D. Synthesis of mannosyl-C-heptylamides

##STR00271##

a) ATMS, Et.sub.2O BF.sub.3, ACN, 0 C..fwdarw.rt. b) Grubb's 2 generation cat., DCM, reflux. c) H.sub.2, Pd(OH).sub.2, MeOH. d) NaN.sub.3, DMF, 80 C. e) Carboxylic acid, HOBt, DIC, Ph.sub.3P, THF, 0 C..fwdarw.rt. f) NaOMe, MeOH.

[1227] Mannosyl-C-heptylamides are obtained from compound 23 through reaction with allyl trimethylsilane (ATMS, step a)), olefin metathesis (step b)), hydrogenation (step c)), displacement of the mesylate using sodium azide (step d)), Staudinger-amide coupling (step e)) and deprotection (step f)).

##STR00272##

a) .sup.tBuOK, THF, 0 C. b) MsCl, Et.sub.3N, DMAP, DCM, 0 C..fwdarw.rt. c) 17, Grubbs catalyst second generation, DCM, 43 C. d) i. H.sub.z, Pd/C, MeOH. ii. NaN.sub.3, DMF, 80 C. e) i. Ph.sub.3P, THF-H.sub.2O, 60 C. ii. Isobutyric chloride, DMAP, DCM, rt. f) i. NaOMe, MeOH, rt, ii. Amberlite IR120 (H).

Example 33

Compound 17

[1228] To a solution of 7-bromo-1-heptanol (2.00 mg, 8.439 mmol) in dry THF (240 mL), cooled at 0 C., was added tBuOK (2.08 g, 18.565 mmol). After stirring at 0 C. for 30 min, 10 ml of H.sub.2O were added and the solvent was evaporated in vacuo.

[1229] To a solution of the crude in dry DCM (40 ml) was added MsCl (715 l, 9.283 mmol), Et.sub.3N (1.76 ml, 12.659 mmol) and DMAP (100 mg). The mixture was stirred for 3 h at rt, washed with saturated solution of NaHCO.sub.3, concentrated under vacuum and the crude was purified by silica gel column chromatography (Hexanes/EtOAc, 70:30) to give the product 17 (1.35 g, 7.089 mmol, 84%) as a colorless oil.

[1230] .sup.1H NMR (300 MHz, CDCl.sub.3): =1.43 (4H, m), 1.76 (2H, m), 2.07 (2H, m), 3.00 (3H, s, MsO), 4.22 (2H, t, J=6.5 Hz), 4.93-5.04 (2H, m, CH.sub.2-alkene), 5.79 (1H, m, CH-alkene).

[1231] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =24.8 (CH.sub.2), 28.2 (CH.sub.2), 28.9 (CH.sub.2), 33.4 (CH.sub.2), 37.3 (CH.sub.2), 70.0 (CH.sub.3, MsO), 114.7 (CH.sub.2, CH.sub.2-alkene), 138.4 (CH.sub.2, CH-alkene).

[1232] MS (CI, NH.sub.3): m/z: [M+NH.sub.3].sup.+ 210

Example 34

Compound 19

[1233] The Grubbs second-generation catalyst (206 mg, 0.242 mmol, 10% mol) was added under argon to a mixture of terminal alkenes 18 (as described by Pawel et al., J Am. Chem. Soc. 2008, 130, 2928-2929; 900 mg, 2.421 mmol) and 17 (1.15 g, 6.053 mmol) in deoxygenated dry DCM (36 ml). The resulting solution was stirred at reflux for 8 h. Removal of the solvent in vacuo gave a brown oil, which could be purified by silica gel column chromatography (Hexanes/EtOAc, 80:20) to give the product 19 (703 mg, 1.304 mmol, Z/E: 8/2, 54%) as a colorless oil.

[1234] .sup.1H NMR (300 MHz, CDCl.sub.3): major isomer 1.39 (4H, m), 1.74 (2H, m), 2.02 (3H, s, AcO), 2.06 (3H, s, AcO), 2.08 (2H, m), 2.09 (3H, s, AcO), 2.12 (3H, s, AcO), 2.42 (2H, m), 3.00 (3H, s, MsO), 3.89 (1H, m, H-5), 3.97 (1H, m, H-1), 4.09 (1H, dd, J.sub.6a,6b=12.1 Hz, J.sub.6a,5=2.9 Hz, H-6a), 4.22 (2H, t, J=6.6 Hz, H-7), 4.32 (1H, dd, J.sub.6b,6a=12.1 Hz, J.sub.6b,5=6.0 Hz, H-6b), 5.18-5.28 (3H, m), 5.37 (1H, m, alkene), 5.57 (1H, m, alkene).

[1235] MS (CI, NH.sub.3): m/z: [M+NH.sub.3].sup.+ 554

[1236] HRMS (ESI): m/z calcd for C.sub.23H.sub.36O.sub.12SNa [M+Na].sup.+: 559.1819, found: 539.1807.

Example 35

Compound 20

[1237] The mixture Z/E of the metathesis product 19 (458 mg, 0.854 mmol) and 10% palladium on carbon (80 mg) in MeOH (15 mL) were stirred under a hydrogen atmosphere (1 atm) at room temperature for 4 h. The reaction mixture was filtered through a pad of Celite and the solvent was evaporated in vacuo.

[1238] A solution of the crude in DMF (26 mL) was added NaN.sub.3 (83 mg, 1.280 mmol) and the resulting mixture was stirred at 70 C. overnight. The mixture was diluted with Et.sub.2O and washed with H.sub.2O and brine. The crude was purified by silica gel column chromatography (Hexanes/EtOAc, 70:30) to give the azide 20 (407 mg, 0.837 mmol, 98%) as a colorless oil.

[1239] [].sub.D=+86.5 (c=0.91 in CHCl.sub.3)

[1240] .sup.1H NMR (300 MHz, CDCl.sub.3): 1.27-1.46 (10H, m), 1.59 (3H, m), 1.77 (1H, m), 2.02 (3H, s, AcO), 2.05 (3H, s, AcO), 2.10 (3H, s, AcO), 2.13 (3H, s, AcO), 3.26 (2H, t, J=6.9 Hz, H-8), 3.84 (1H, ddd, J.sub.5,4=8.8 Hz, J.sub.5,6a=6.1 Hz, J.sub.5,6b=2.7 Hz, H-5), 3.94 (1H, ddd, J=10.0 Hz, J=4.6 Hz, J=2.6 Hz, H-1), 4.09 (1H, dd, J.sub.6a,6b=12.2 Hz, J.sub.6a,5=2.8 Hz, H-6a), 4.30 (1H, dd, J.sub.6b,6a=12.2 Hz, J.sub.6b,5=6.1Hz, H-6a), 5.16-5.26 (3H, m, H-2, H-3, H-4).

[1241] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =20.3 (CH.sub.3, AcO), 20.36 (CH.sub.3, AcO), 20.40 (CH.sub.3, AcO), 20.44 (CH.sub.3, AcO), 25.6 (CH.sub.2), 26.8 (CH.sub.2), 28.7 (CH.sub.2), 28.9 (CH.sub.2), 29.1 (CH.sub.2), 29.3 (CH.sub.2), 29.6 (CH.sub.2), 51.3 (CH.sub.2, C-8), 62.7 (CH, C-6), 67.6 (CH), 69.6 (CH), 71.1 (CH), 71.2 (CH), 75.0 (CH, C-1), 169.4 (C, AcO), 169.8 (C, AcO), 169.9 (C, AcO), 170.0 (C, AcO).

[1242] MS (CI, NH.sub.3): m/z: [M+NH.sub.3].sup.+ 503

[1243] HRMS (ESI): m/z calcd for C.sub.22H.sub.35N.sub.3O.sub.9N.sub.3Na [M+Na].sup.+: 508.2265, found: 508.2257

Example 36

Compound 21

[1244] A mixture of 20 (350 mg, 0.720 mmol) and 10% palladium on carbon (35 mg) in MeOH (7 mL) were stirred under a hydrogen atmosphere (1 atm) at room temperature for 10 h. The reaction mixture was filtered through a pad of Celite and the solvent was evaporated in vacuo. To a solution of the amine crude in dry DCM (14 ml) was added isobutyric chloride (114 1.080 mmol) and DMAP (264 mg, 2.160 mmol) at 0 C. under N.sub.2 atmosphere. The mixture was stirred for 5 h at rt, concentrated under vacuum and the crude was purified by silica gel column chromatography (DCM/MeOH, 90:10) to give the product 21 (253 mg, 0.477 mmol, 66% from 20) as a colorless oil.

[1245] [].sub.D=+59.2 (c=0.75 in CHCl.sub.3)

[1246] .sup.1H NMR (300 MHz, CDCl.sub.3): =1.11 (6H, d, J=6.9 Hz, 2CH.sub.3-isobutyric acid), 1.22-1.57 (13H, m), 1.74 (1H, m), 1.98 (3H, s, AcO), 2.02 (3H, s, AcO), 2.06 (3H, s, AcO), 2.10 (3H, s, AcO), 2.29 (1H, m, CH, isobutyric acid), 3.19 (2H, m, H-8), 3.77 (1H, m, H-5), 3.87 (1H, ddd, J=10.0 Hz, J=4.6 Hz, J=2.6 Hz, H-1), 4.10 (1H, dd, J.sub.6a,6b=12.1 Hz, J.sub.6a,5=2.6 Hz, H-6a), 4.04 (1H, dd, J.sub.6b,6a=12.1 Hz, J.sub.6b,5=5.9 Hz, H-6a), 5.09-5.20 (3H, m, H-2, H-3, H-4), 5.53 (1H, bs, NH).

[1247] .sup.13C NMR(100.6 MHz, CDCl.sub.3): =19.6 (2CH.sub.3, isobutyric acid), 20.2 (CH.sub.3, AcO), 20.65 (CH.sub.3, AcO), 20.67 (CH.sub.3, AcO), 20.9 (CH.sub.3, AcO), 25.2 (CH.sub.2), 26.7 (CH.sub.2), 28.3 (CH.sub.2), 28.9 (CH.sub.2), 29.0 (CH.sub.2), 29.2 (CH.sub.2), 29.6 (CH.sub.2), 35.6 (CH, isobutyric acid), 39.2 (CH.sub.2, C-8), 62.6 (CH, C-6), 66.8 (CH), 69.0 (CH), 69.9 (CH), 70.8 (CH), 75.2 (CH, C-1), 169.6 (C, AcO), 169.9 (C, AcO), 170.3 (C, AcO), 170.6 (C, AcO), 176.8 (C, amide).

[1248] MS (CI, NH.sub.3): m/z: [M+H].sup.+ 530

[1249] HRMS (ESI): m/z calcd for C.sub.26H.sub.440.sub.10N [M+H].sup.+: 530.2959, found: 530.2955.

Example 37

Compound 22

[1250] According to the general procedure B, using the amide 21 (50 mg, 0.094 mmol) as starting material, the derivative 22 was obtained after lyophilization (33 mg, 0.091 mmol, 97%) as an amorphous white solid.

[1251] [a].sub.D=+26.6 (c=0.81 in MeOH).

[1252] .sup.1H NMR (300 MHz, MeOD): =1.12 (6H, d, J=6.9 Hz, 2CH.sub.3-isobutyric acid), 1.31-1.79 (14H, m), 2.44 (1H, m, CH-isobutyric acid), 3.17 (2H, q, J=6.3 Hz, C-8), 3.42 (1H, m, H-5), 3.61-3.89 (6H, m), 7.85 (1H, bs, NH).

[1253] .sup.13C NMR (100.6 MHz, MeOD): =19.9 (2CH.sub.3, 2CH.sub.3-isobutyric acid), 26.9-30.5 (7CH.sub.2), 36.3 (CH, CH-isobutyric acid), 40.2 (CH.sub.2, C-8), 63.1 (CH, C-6), 69.3 (CH), 72.8 (CH), 73.1 (CH), 75.5 (CH), 78.9 (CH, C-1), 179.9 (C, amide).

[1254] MS (CI, NH.sub.3): m/z 362 [M+H].sup.+

[1255] HRMS (MALDI, DHB): m/z calcd for C.sub.18H.sub.35O.sub.6NaN [M+Na].sup.+: 384.2357, found: 384.2354.

Example 38

Adhesion Ability of Adherent-Invasive E. coli to Intestinal Epithelial Cells in Presence of Monovalent Compounds: Pre-, Co- and Post-Incubation Experiments.

Bacterial Strain and Cell Line

[1256] E. coli strain LF82 was isolated from a chronic ileal lesion of a patient with Crohn's disease (CD). Bacteria were grown routinely in Luria-Bertani (LB) broth overnight at 37 C. Intestinal epithelial cells T84 derived from colonic adenocarcinoma were maintained in an atmosphere containing 5% CO2 at 37 C. in DMEM/F12 (50/50) medium supplemented with 10% (v/v) heat-inactivated fetal calf serum (FCS), 1% L-glutamine, 100 000 U.1.sup.1 penicillin, 100 mg.1.sup.1 streptomycin, 25 g.1.sup.1 amphotericin B.

Adhesion Ability of Adherent-Invasive E. coli to Intestinal Epithelial Cells in Presence of Monovalent Compounds.

[1257] T84 were seeded in 48-well plates at a concentration of 1.510.sup.5 cells per well and grown for 48 h. AIEC LF82 bacteria were incubated 1 h with monovalent compounds prior the cell infection (pre-incubation protocol) or they were added simultaneously onto the cells (co-incubation protocol) in complete medium without antibiotics, containing heat inactivated fetal calf serum (FCS). Monovalent compound 10 was tested at a dose of 100; 10; 1 (and 0.1) M, compound 5 was tested at a dose of 500; 100; 10 and 1 M and D-mannose was tested at a dose of 10 000; 1 000; 100 or 10 M. Cells were infected with AIEC LF82 bacteria at a multiplicity of infection (MOI) of 10 bacteria per cell for 3 h at 37 C.

[1258] For the post-incubation protocol, monovalent compounds (same doses than in the pre- and co-incubation assays) were incubated with cells for 3 h after bacterial infection. A washing step was realized before this post-incubation to eliminate non-adherent bacteria.

[1259] Monolayers were washed in phosphate-buffered saline (PBS; pH 7.2) and cells were then lysed with 1% Triton X-100 in deionized water. Samples were diluted and plated onto Luria Bertani agar plates to determine the number of colony-forming units (CFU) recovered from the lysed monolayers. Results were expressed as percentages of residual adhesion, considering adhesion level of AIEC LF82 without mannosides treatment as 100%.

Results

[1260] D-mannose, compounds 5 and 10 were assessed as putative inhibitors to compete the interaction of CEACAM6 expressed by T84 intestinal epithelial cells with the adhesin FimH of AIEC bacteria following three different protocols: pre-, co-and post-incubation experiments (FIG. 1).

[1261] For co-incubation experiments, results clearly indicated that monovalent compound 10 possessed the best inhibitory effect, with a 50-fold increased potency in comparison with the compound 5 and a 100-fold increased potency in comparison with D-Mannose. Significant decreases in the bacterial adhesion levels were obtained at 10 M for compound 10, at 500 M for compound 5 and at 1 000 M for D-Mannose. Using the pre-incubation protocol, D-mannose showed a significant inhibitory effect on the bacterial adhesion at 100 M, whereas similar results than those observed with the co-incubation protocol were obtained for compounds 5 and 10. Finally, in post-incubation experiments, D-mannose decreased adhesion at a high dose of 10 mM, compound 5 was not able to decrease bacterial adhesion even at 500 M, and compound 10 showed a significant inhibitory effect at 100 M. These data indicated that monovalent compound 10 is a good inhibitor to detach bacteria adhering to intestinal epithelial cells at a dose of 100 M.

Example 39

Adhesion Ability of Adherent-Invasive E. coli Strains to Intestinal Mucosa of Transgenic Mice Expressing CEACAM6 in the Presence of Monovalent Compounds

Bacterial Strain and Transgenic Mouse Model

[1262] E. coli strain LF82 was isolated from a chronic ileal lesion of a patient with Crohn's disease (CD). Bacteria were grown routinely in Luria-Bertani (LB) broth overnight at 37 C.

[1263] The transgenic mouse model CEABAC10 expressing the human CEACAM6 protein is available in the UMR Inserm/Universit d'Auvergne 1071 led by Professor Arlette Darfeuille-Michaud at Clermont-Ferrand. This model is particularly suitable to mimic the abnormal colonization of gut mucosa by AIEC bacteria through the interaction of CEACAM6 abnormally expressed in the ileal mucosa of Crohn's disease and FimH adhesin of AIEC.

Adhesion Assays of Adherent-Invasive E. coli Strains to Colonic Loops from CEABAC10 Mice in Presence of Monovalent Compounds.

[1264] Three colonic loops were performed in anesthetized CEABAC10 mice. A volume of 100 l of a bacterial suspension containing 2.510.sup.7 bacteria/mL in the presence or absence of monovalent compounds was injected into the loops (here, compound 10 at a concentration of 100 M). After an incubation period of 4 h, mice were euthanized and each loop was longitudinally opened, extensively washed in phosphate buffer and homogenized to numerate adherent LF82 bacteria. Bacterial adhesion levels were expressed as colony forming units (CFU) per gram of colonic tissue in the FIG. 2 (100% corresponds to the LF82 adhesion in absence of any compound).

Results

[1265] A-two fold decrease in the number of LF82 bacteria adhering to colonic mucosa was observed in the presence of the monovalent compound 10 at a concentration of 100 M (FIG. 2).

Example 40

Effect of Orally Administered Monovalent Compounds to Adherent-Invasive E. coli LF82-infected transgenic mice expressing CEACAM6

Bacterial Strain and Transgenic Mouse Model

[1266] E. coli strain LF82 was isolated from a chronic ileal lesion of a patient with Crohn's disease (CD). Bacteria were grown routinely in Luria-Bertani (LB) broth overnight at 37 C. The transgenic mouse model CEABAC10 expressing the human CEACAM6 protein is available in the UMR Inserm/Universit d'Auvergne 1071 led by Professor Arlette Darfeuille-Michaud at Clermont-Ferrand. This model is particularly suitable to mimic the abnormal colonization of gut mucosa by AIEC bacteria through the interaction of CEACAM6 abnormally expressed in the ileal mucosa of Crohn's disease and FimH adhesin of AIEC.

AIEC colonization assessment in CEABAC10 mice treated with monovalent compounds.

[1267] To mimic curative treatment, compounds were analyzed for their anti-adhesive effect on a pre-established LF82 colonization in CEABAC10 mice. CEABAC10 mice were given 0.5% of DSS in drinking water. Two days later, mice were treated per os with streptomycin sulfate, 5 mg/mouse. Twenty four hours later, (corresponding to day 0), a five-hour culture of AIEC LF82 bacteria in LB broth was concentrated to reach 510.sup.9 bacteria/mL and was administered by gavage 2 h after the intragastric administration of cimetidine at 50 mg/kg in order to ablate gastric secretion. Oral administration of monovalent compounds at a range from 1 to 1000 g/mouse (=0.04 to 40 mg/kg) was realized 2 h after LF82 infection. A second administration of the compounds was realized 18 h later (cimetidine was also given 2 h before administration of the compounds). Body weight and signs of colitis were followed for 4 days. Stools were collected from day 1 to day 4 post-infection to assess bacterial colonization. Mice were euthanized at day+4 and the entire intestine was collected to assess the number of AIEC bacteria associated with the gut mucosa, to measure pro-inflammatory cytokine secretion, to measure myeloperoxidase activity as indicator of neutrophil infiltration in the intestinal tissue, to determine the disease activity index and to estimate histological damages of the mucosa.

[1268] Similar protocol was realized in testing a prophylactic administration of the compounds (administration of similar doses of monovalent compounds 5 h before infection). Compounds were compared for their efficacy, depending on the dose and on the preventive or curative effect. To analyze whether the inhibitory effects could be related to toxicity effects, the absence of cell death of intestinal epithelial cells or bacteria was assessed at the highest dose of each compound.

[1269] CEABAC10 mice were infected with 10.sup.9 bacteria at day 0 (DO) and then orally treated two times with 10 mg/kg of compounds 5 and 10 (curative treatment). Body weight was followed during 3 days before LF82 infection and until day 4 post-infection. Bacterial loads in feces and signs of colitis were followed at day 1, 3 and 4 after infection as well as the severity of colitis, assessed by establishment of the Disease Activity Index score (DAI). The numbers of bacteria associated to the intestinal mucosa were assessed at day 4 post-infection.

[1270] Intestinal tissues were sampled to measure the levels of pro-inflammatory cytokines and to analyze damages of mucosa (HES staining of colonic slices). Finally, spleen were collected and weighted.

Results

[1271] The body weight mean of the LF82-infected mice strongly decreased between day 0 and day 4 post-infection, compared to the non-infected group. LF82-infected mice treated with compound 5 or 10 did not show any decrease in the body weight (FIG. 3). Compared to LF82-infected mice, LF82-infected mice treated with monovalent compounds 5 or 10 showed very low DAI scores at day 3 and 4 post-infection, similar to that of non-infected mice (FIG. 4D). The LF82 colonization levels were strongly decreased in the feces of LF82-infected mice treated with 5 and 10 (with less than 10.sup.4 bacteria/g of feces), in comparison with LF82-infected but non treated mice (more than 10.sup.6 bacteria/g of feces) (FIGS. 4A, 4B and 4C). Similar decreased colonization in the presence of compounds 5 and 10 was observed for the number of bacteria associated to the ileum and the colon (0 CFU/g of intestinal tissue) compared to 510.sup.3 and 110.sup.4 CFU/g of tissue for ileum and colon, respectively, in the absence of any compounds (FIG. 5). Compared to the non-infected mice, increased spleen weight was observed in LF82-infected mice. This was no longer observed when mice were treated with monovalent compounds 5 and 10 (FIG. 6). Finally, in that infection model, LF82 was able to increase the levels of pro-inflammatory cytokines IL-23, KC and TNF- secreted, compared to non-infected mice. When mice were treated with monovalent compound, 5 and 10 the levels of pro-inflammatory cytokines secreted were decreased compared to non-treated mice. Decreases were significant for the three IL-23, KC and TNF- cytokines in the presence of compound 10 but not in the presence of compound 5 (FIG. 7).

Example 41

In Vitro Screening of Anti-FimH Molecules

[1272] Molecules were screened for their inhibition effect on the adhesion of the AIEC LF82 strain to intestinal epithelial T84 cells.

[1273] The molecules tested were the O-glycosides 10 and 5, the S-glycoside 16 and the C-glycoside 22.

Post-Incubation Protocol with Undifferentiated T84 Cells:

[1274] The AIEC LF82 strain was incubated with T84 cells and then the tested molecule was added.

[1275] The Protocol is as follows: [1276] Cells: T84, 48h culture, in 48 wells plate at 1.510.sup.5 cells/well; [1277] Bacteria: AIEC LF82 strain, Culture ON; [1278] Inhibitor compounds in mother solutions (10, 20, 50 or 100 mM); [1279] Measure the OD(620) of the bacterial culture; [1280] Prepare the bacterial suspension at 610 .sup.6 bact/mL in DMEM/F12/SVF dec 10% medium; [1281] Wash twice the cellular layer with PBS; [1282] Add 250 l/well of bacteria suspension, id. 1.510.sup.6 bact/well (MOI=10); [1283] Incubate 3 hours at 37 C.; [1284] Prepare inhibitor compounds at the wished final concentration in DMEM/F12/SVF dec 10% medium and filtrate at 0.2 l filter; [1285] Wash 5 times the cellular layer with PBS; [1286] Add 250 L of inhibitor compounds/well; [1287] Incubate 3 hours at 37 C.; [1288] Wash 5 times the cellular layer with PBS; [1289] Add 250 L of Triton X-100 at 1%, incubate 5 min at room temperature then add Triton in each well; [1290] Take the entire content of each well and transfer it in an Eppendorf tube of 1.5 mL; [1291] Perform serial dilutions in physiological water: 50 l of sample in 450 l of physiological water;

[1292] (NB: Prepare physiological water +2% D-mannose if difficulty met to get isolated colonies) [1293] Spread 25 l of dilution on LB-agar gelose; [1294] Incubate overnight at 37 C.

[1295] For this experiment, all compounds have been tested at a final concentration of 100 M.

Criteria of Evaluation:

[1296] The criteria of evaluation is the residual adhesion (level of colonization/decolonization of AIEC measured on cells) expressed in percentage.

Results: Dose Effect with Tested Molecule at Different Concentrations:

[1297] Pre incubation experiments and post incubation experiments (FIG. 8) provide consistent results with respect to 10/22 and 5/16.

Example 42

In Vivo Testing of Anti-Adhesive Effect of Molecules on AIEC LF82 Colonization in CEABAC10 Mice

[1298] Molecules tested: 10 (1 mg/kg and 10 mg/kg), 22 (10 mg/kg), 16 (10 mg/kg), 5 (10 mg/kg) The aim was to test different compounds given per os to CEABAC10 mice infected by AIEC LF82 strains by assessing their ability to decrease bacterial colonization and related colitis.

Protocol:

[1299] Mice CEABAC10 (8 weeks-old males) were given DSS 0.5% in water for all the time of experiment. [1300] Two days later, mice were treated p.o. with Streptomycin sulfate, 5 mg/mouse (in water). [1301] The following day (=DO), LB broth was inoculated (1/100.sup.th) with an ON culture of AIEC LF82 and incubated at 37 C. with shaking in order to obtain a DO=0.5 or 0.6 maximum. Bacteria were concentrated at 1.510.sup.10 bacteria/mL and 0.2 mL was administered intra-gastrically to mice (=310.sup.9 bact/mouse) 2 h after oral administration of cimetidine at 50 mg/kg in order to ablate gastric secretion (6.25 mg/mL in water, 0.2mL/mouse). [1302] Tested molecules were orally given twice at a dose of 250 g/mouse (=10 mg/kg) or 25 g/mouse (=1 mg/kg) in PBS, 2 h and 18 h after infection (cimetidine was given 2 h before each administration of the compounds). [1303] Body weight were followed for 3 (for ANRS) to 4 (for ANR3) days. [1304] Stools were collected at day+1 (ANR3 and ANRS), day+3 post-infection (ANRS), day+4 post-infection (ANR3) to assess bacterial colonization. [1305] Mice were euthanized at day+4 for ANR3 and day+3 for ANRS and entire intestine was collected to assess the bacterial colonization at the mucosa (ileum+colon), to measure pro-inflammatory cytokine secretions, to assess the neutrophil infiltration into the tissues (myeloperoxidase activity), to assess the disease activity index. [1306] ANR3: Groups of male mice (32 mice in total): [1307] A. Non-infected (NI) mice; n=8 [1308] B. LF82-Infected mice without treatment; n=8 [1309] C. LF82-Infected mice+10 at 10 mg/kg; n=8 [1310] D. LF82-Infected mice+5 at 10 mg/kg; n=8 [1311] ANRS: Groups of male mice (48 mice in total): [1312] E. Non-infected (NI) mice; n=12 [1313] F. LF82-Infected mice without treatment; n=12 [1314] G. LF82-Infected mice+22 at 10 mg/kg; n=12 [1315] H. LF82-Infected mice+16 at 10 mg/kg; n=12

Criteria of Evaluation:

[1316] 1. Body weight

[1317] 2. Disease activity index (DAI)

[1318] 3. Bacterial colonization in stools

[1319] 4. Bacterial colonization at the mucosa

[1320] 5. Pro-inflammatory cytokine secretions

[1321] 6. Neutrophil infiltration into the tissues (myeloperoxidase activity)

Results:

[1322] The evolution of the weight of CEABAC10 transgenic mice uninfected or infected with AIEC LF82 was followed after administration of various molecules to be tested (FIG. 9). Infection of mice with MEC LF82 leads to a decrease in the weight of mouse. The administration of molecules 10, 22 and 16 prevents weight loss induced by infection with the AIEC LF82 strain (FIGS. 9A and 9B). This observation correlates with decreased of disease activity index (DAT score) 3 days after infection following administration of molecules 10, 22 and 16 in mice infected with AIEC LF82 (FIG. 10). To assess the ability of molecules to reduce the colonization of the intestinal mucosa by AIEC strains, the amount of AIEC bacteria present in the feces, which reflects the amount of AIEC bacteria associated with intestinal mucosa, was measured. As control, one day post-infection, the amount of MEC bacteria present in feces was comparable irrespective of the administration of molecules tested, indicating a similar level of colonization in all the batches from the experiment. Interestingly, administration of molecule 22 lead to a decrease in the amount of AMC LF82 bacteria in the feces of infected mice at 3 and 4 days post-infection, showing the effectiveness of these molecules to decrease the ability of the MEC LF82 to colonize the mouse intestine (FIG. 11). In addition, the count of ALEC LF82 bacteria associated with ileal or colonic mucosa at 4 day post-infection shows that the molecules 22 and 16 decolonized AIEC bacteria very effectively at the ileal and colonic mucosa (FIG. 12), Various inflammatory parameters were measured at 3 or 4 days post-infection. First, the myeloperoxidase (MPO) activity, which reflects the infiltration of the intestinal mucosa by neutrophils, was measured. Interestingly, administration of molecules 10, 5, 22 led to a decrease in MPO activity (FIG. 13). In addition, the quantification of the pro-inflammatory cytokines IL23 (FIG. 14) and IL-Ibeta (FIG. 15) was performed at the level of mucosa from infected mice. The administration of the molecules 10, 5, 16 resulted in a reduction of the cytokine IL23 level and administration of the molecules 10, 5 led to a decrease in the release of IL I-beta.

[1323] All of these in vivo results obtained in the transgenic mouse model CEABAC 10 infected with AIEC LF82 shows that different molecules tested either reduced the activity of the disease (weight and score DAT), the level of colonization of mucosa or inflammatory parameters (MPO activity and production of pro-inflammatory cytokines), suggesting that these molecules are potentially useful in the treatment of Crohn's disease patients colonized by AIEC strains.

Example 43

Ex Vivo Protocol

[1324] The ex vivo model of explant cultures from human colonic mucosa is used to examine the interactions of the AIEC strain LF82 with human colonic mucosa (controls) cells and the decolonization of AIEC from mucosa cells thanks to the FimH antagonists molecules (treated).

[1325] Human Colonic mucosa explants. The mucosa is carefully stripped from the underlying compartment. Fragments of 40 mg are maintained in culture overnight in RPMI-BSA 0.01% supplemented with gentamicin to get rid of commensal bacteria, and fungizone washed twice in RPMI, and then incubated for 4 h with or without bacterial cultures (LF82-GFP) in 2 ml culture medium without antibiotics. The explants are maintained at 37 C. in a 95% oxygen, 5% carbon dioxide humid atmosphere on a rocking platform at low speed. In each experiment, at least three explants are cultured for each condition. The supernatants are centrifuged and aliquots are stored at 80 C. for further analysis.

[1326] Bacterial strains and media. The prototype AIEC strain LF82-GFP is used (UMR 1071 Inserm/Universite d'Auvergne, Clermont-Ferrand, France). The strains are stored at 20 C. in cryotubes. Before the experiments, the bacteria are cultivated on TS agar at 37 C. for 24 h after thawing. For each experiment, bacteria are subcultured in LB broth at 37 C. for 18h with shaking. The bacteria are then centrifuged for 10 min at 800g. The pellet is washed twice with sterile PBS, and the suspension is adjusted to 0.510.sup.8 or 0.510.sup.9 bacteria per milliliter, in culture medium (RPMI/BSA 0.01% without antibiotics).

[1327] The explant cultures, left to stabilize overnight in culture medium with antibiotics, were co-incubated with LF82-green fluorescent protein (GFP) (10.sup.8 or 10.sup.9 bacteria per explant) without antibiotics for 4 h. LF82 bacteria, detected by immunoperoxidase using an anti-GFP antibody on paraffin sections, are found adhering to the apical pole of a few epithelial cells of the surface and crypt base, scattered or sometimes focally clustered.

Example 44

Pharmacokinetic Study Following Administration of 2 Compounds by Oral and Intravenous Administration to Male Sprague Dawley Rats

[1328] These in vivo and analytical experiments are conducted to: [1329] Estimate the plasma concentration level after oral and intravenous administrations of 2 compounds to male Sprague Dawley rats; [1330] Calculate the bioavailability; [1331] Estimate the amount of unchanged compounds in the faeces.

[1332] Two substances are tested (previously stored at room temperature in the dark).

[1333] For analysis, the molecules are dissolved in DMSO at 1 mg/mL.

TABLE-US-00001 Compounds Weight tube (mg) 10, 22 2 * 1 mg for the analytical part 15 mg for the in vivo part

1. Analytical Test

[1334] Before the beginning of the in vivo part, an analytical test for each compound are performed in the two matrices.

[1335] The molecular and daughter ions are selected for each compound by direct infusion into the MS-MS system.

[1336] At least 8 point calibration standards are run using standard conditions which consist to LC-MS/MS system with C18 column after precipitation of proteins before the start of the analytical test.

[1337] Blank rat faeces are homogeneized with 3 volumes of UHQ water until obtention of a paste.

[1338] Then 100 L of the homogenate are spiked with the molecules and precipitated with 300 L of acetonitrile.

[1339] For the plasma, 100 L of blank rat plasma are directly spiked with the compounds before being precipitate with 300 L of acetonitrile.

[1340] The corresponding correlation coefficient (r) is calculated and should be higher than 0.75 to continue with the in vivo test.

[1341] The concentration ranges tested are: [1342] 0.5 ng/mL to 1000 ng/mL for plasma, [1343] 4 to 2000 ng/g for faeces, corresponding to 1 to 500 ng/mL of faeces homogenate.

2. In-Life Part

[1344] 2.1. Characteristics, Housing and Handling of Animals

[1345] 30 male Sprague Dawley rats around 6-7 week old are used.

[1346] At reception, the animals are housed in makrolon cages with stainless steel wire lids with catches. A label on each cage indicates the reception date, the rat strain, sex and weight.

[1347] Temperature and humidity are continually monitored. The animal room conditions is kept as follows: [1348] Temperature: 22 C.2 C. Exceptionally, upper or lower values can be tolerated. [1349] Light/dark cycle: 12 h/12 h (07:00 h-19:00 h).

[1350] After administration and over the experiment duration, the animals are placed individually in metabolic cages (tecniplast).

[1351] Animals have free access to food and water during the experiment.

[1352] 2.2. Design

TABLE-US-00002 Volume of Dose Concen- administration Compounds Route vehicle (mg/kg) tration (mL/kg) 10, 22 IV 100% DMSO 1 1 mg/mL 1 PO 100% DMSO 10 2 mg/mL 5

[1353] 2.3. Sampling

[1354] For each test substance

TABLE-US-00003 Admin- Blood Faeces istration sampling sampling Molecule route Rat name Time Time 10, 22 IV IV1, IV2, 5 min 0-24 h IV3 30 min 2 h 6 h 24 h PO PO4, PO5, 30 min 0-24h PO6 1 h 2 h 6 h 24 h

[1355] After administration, the animals are placed in individual metabolic cages in order to collect faeces samples during 24 hours.

[1356] 2.4. Blood Sampling

[1357] At prescribed times, blood will be collected. Animals are briefly anaesthetised with Isoflurane using an anaesthetic system (quipement Vtrinaire Minerve) during blood samplings. [1358] Site of collection: sinus retro-orbital using a capillary tube [1359] Volume of blood collected: 0.3 mL per time-point [1360] Anticoagulant: Heparin Lithium

[1361] Exact sampling times are noted for each blood sampling.

[1362] Blood samples are centrifuged at 2500 rpm at +4 C. (between 0 and 9 C.), the plasma is removed and placed into labelled polypropylene tubes. Individual plasma samples is stored frozen at 20 C. (target temperature) until analysis.

3. Analysis

[1363] 3.1. Analysis of plasma samples

[1364] 100 L of the plasma sample are taken and 300 L of acetonitrile are added. After protein precipitation, analysis are performed using LC-MS/MS determination according to previous analytical test results.

[1365] 3.2. Analysis of Faeces Samples

[1366] Faeces samples are collected over the 24 hours of the experiment.

[1367] They are precisely weighed and 3 volumes of UHQ water are added.

[1368] The mixture is homogeneized until obtention of a paste.

[1369] Then 100 L of the homogenate are taken and extracted with 300 L of Acetonitrile.

[1370] Analysis is performed using LC-MS/MS determination according to previous analytical test results.

[1371] 3.3. Determination of the Concentrations

[1372] Concentrations of the samples are calculated directly from chromatograms after automatic integration by Analyst 1.5.1 and expressed as ng/mL.

[1373] Mean plasma concentrations are calculated (when calculable, i.e. n2) using individual concentration and are expressed with the corresponding standard deviation value and

[00001] $(with .Math. .Math. CV .Math. .Math. (%) = \frac{SD}{Cmean} 100) .$

variation coefficient (when calculable, i.e. n 3)

[1374] The individual plasma concentrations are tabulated for each rat and scheduled sampling time. Concentrations below the LLOQ are indicated by BLQ. All BLQ concentrations are substituted by zero for calculation of the descriptive statistics of the concentrations.

4. Results

[1375] The results is provided with plasma concentration/time curves, as well as the tabulated concentrations results obtained for each plasma and faeces time point.

[1376] Estimation of PK parameters is performed using Kinetica (Version 4.3Thermo Electron

[1377] CorporationPhiladelphiaUSA). An independent model method is used. The following parameters are estimated: [1378] Cmax (ng/mL): maximal plasma concentration [1379] Tmax (h): first time to reach Cmax [1380] AUC.sub.t (ng/mL*h): area under the plasma concentration-time curve from administration up to the last quantifiable concentration at time t [1381] Absolute bioavailability:

[00002] $F .Math. .Math. (%) .Math. = \frac{AUC .Math. .Math. PO / dose .Math. .Math. PO}{AUC .Math. .Math. IV / dose .Math. .Math. IV} * 100$

Example 45

Testing of Resistance to Mannosidases

[1382] Compounds are testing for degradation by intestinal enzymes like mannosidases, known to preferentially induce breakage between mannose and O-linkage. To avoid such degradation, several analogues have been designed, for example 22 (CH2-analogue of 10), 16 (S analogue of 5).

[1383] Each compound is incubated with mannosidase and with or without inhibitors of mannosidase. Mass spectrometry experiments are performed to detect native and degraded compounds.

Example 46

In Vitro Toxicity Studies

[1384] The cytotoxic activity of compounds 10 and 22 against normal cell lines using MTS assay was determined.

Materials and Methods

[1385] Compounds 10 and 22 were extemporaneously dissolved at 100 mM in water to obtain a stock solution.

[1386] The final concentrations of compounds 10 and 22 were 100 nM, 1 M, 10 M, 100 M and 1 mM within wells.

[1387] The cell lines that were used are detailed in the table hereafter:

TABLE-US-00004 Cell line Type Species Origin HUV-EC-C Umbilical vein Human Millipore endothelial cells CCD-18Co Colon normal Human ATCC CRL-1459 fibroblast MRC-5 Normal ftal Human ATCC CCL-171 lung fibroblast PWR-1E Normal prostate Human ATCC CRL-11611 cells

[1388] The 4 cell lines were plated at optimal density per well in 96-well plates. Plates were incubated at 37 C. for 24 hours before treatment, in drug-free culture medium. Cell lines were then incubated for 96 hours at 37 C. under 5% CO.sub.2 with the 5 concentrations of compounds 10 or 22 in 1:10 dilution steps. Each concentration was done in triplicate. Control cells were treated with vehicle alone (water). At the end of the treatment, the cytotoxic activity of compounds 10 and 22 was assessed by MTS assay.

[1389] The in vitro cytotoxic activity of compounds 10 and 22 was revealed by a MTS assay using tetrazolium compound (MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulfophenyl)-2H-tetrazolium) and an electron coupling reagent named PMS (phenazine methosulfate).

[1390] The dose response for index of cytotoxicity (IC) is expressed as following:

[00003] $IC = \frac{{OD}_{drug - exposed .Math. .Math. wells}}{{OD}_{vehicule - exposed .Math. .Math. wells}} 100$

[1391] The OD values are the mean of 3 experimental measurements. IC.sub.50 represents the drug concentration required to obtain 50% of cellular cytotoxicity. The IC.sub.50 determination values were calculated from semi-log curves.

Results

[1392] Cytotoxicity studies have shown that compounds 10 and 22 do not present an acute toxicity towards the above-mentioned four cell lines.

[1393] The values obtained for Docetaxel (control compound) are consistent with known values of inherent toxicity, thus validating these experiments.

ORALLY AVAILABLE COMPOUNDS, A PROCESS FOR PREPARING THE SAME AND THEIR USES AS ANTI-ADHESIVE DRUGS FOR TREATING E. COLI INDUCED INFLAMMATORY BOWEL DISEASES SUCH AS CROHN'S DISEASE

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