α-O-linked glycoconjugates, methods of preparation and uses thereof

A method of treating cancer in a subject suffering therefrom comprising administering to the subject a therapeutically effective amount of a glycoconjugate having the structure:

• wherein m′, n′ and p′ are integers between about 8 and 20;
• wherein j is an integer between 1 and about 8;
• wherein RV, RA, RB and RC are independently hydrogen, substituted or unsubstituted linear or branched chain lower alkyl or substituted or unsubstituted phenyl;
• wherein RD, RE and RF are independently a carbohydrate domain having the structure:
• wherein a, b, c, d, e, f, g, h, i, x, y and z are each independently 0, 1, 2 or 3, with the proviso that RD, RE, and RF are carbohydrates independently comprised of furanose or pyranose moieties, whereby the sum of b and c is 1 or 2, the sum of d and f is 1 or 2, and the sum of g and i is 1 or 2, and with the proviso that x, y and z are not simultaneously 0;
• wherein R0 is a hydrogen, linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of R1, R2, R3, R4, R5, R6, R7, R8, and R10 is independently hydrogen, OH, ORi, NH2, NHCORi, F, CH2OH, CH2ORi, a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono-, di- or tri)acyloxyalkyl, arylalkyl, or aryl group;
• wherein each occurrence of Ri is independently hydrogen, CHO, CO2Rii, a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl, or aryl group, or a saccharide moiety having the structure:
• wherein Y and Z are independently NH or O;
• wherein k, l, r, s, t, u, v and w are each independently 0, 1 or 2, with the proviso that the v and w bracketed structures represent furanose or pyranose moieties and the sum of l and k is 1 or 2, and the sum of s and u is 1 or 2, and with the proviso that v and w are not simultaneously 0;
• wherein R′0 is hydrogen, a linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of R10, R11, R12, R13, R14 and R15 is independently hydrogen, OH, ORiii1, NH2, NHCORiii, F, CH2OH, CH2ORiii, or a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono- di- or tri)acyloxyalkyl, arylalkyl or aryl group;
• wherein R16 is hydrogen, CO2H, CO2Rii, CONHRii, a substituted or unsubstituted linear or branched chain alkyl or aryl group;
• wherein each ocurrence of Riii is independently hydrogen, CHO, CO2Riv, or a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• and wherein each occurrence of R′ and Riv is independently hydrogen, or a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• with the limitation that each of RD, RE, and RF comprise a carbohydrate domain, or truncated or elongated version thereof, that is present on tumor cells.

A method of treating cancer in a subject suffering therefrom comprising administering to the subject a therapeutically effective amount of a glycoconjugate having the structure:

• wherein each occurrence of RA, RB and RC is independently H or methyl;
• n is 1, 2, 3 or 4;
• j is 1-8;
• t′ is 1-8;
• s′ is 0 or 1, wherein when s′=0, the carrier is a lipid, and when s′=1, the carrier is a protein;
• each occurrence of RD, RE and RF is independently a carbohydrate domain selected from the group consisting of Tn, TF, 2,6-STF, 2,6-STn, 3-Ley, 6-Ley, 3,6-STn, 2,3-ST, a carbohydrate having the structure:
• a carbohydrate having the structure:
• a carbohydrate having the structure:
• wherein j′, k′ and l′ are each independently 0, 1 or 2;
• and a Ley hexasaccharide having the structure:
• wherein j′ and k′ are each independently 0, 1 or 2.

A method of inducing antibodies in a subject, wherein the antibodies are capable of specifically binding with human tumor cells, which comprises administering to the subject an amount of a glycoconjugate effective to induce antibodies which glycoconjugate has the structure:

• wherein the linker is —O—, —NRG—, —NRG(CRHRJ)kNRK—, NRG(CRHRJ)kNRK(C═O)(CRHRJ)kS—, —(CRHRJ)kNRK—, —O(CRHRJ)kNRK—, an oligoester fragment comprising from 2 to about 20 hydroxy acyl residues, a peptidic fragment comprising from 2 to about 20 amino acyl residues, or a linear or branched chain alkyl or aryl carboxylic ester, wherein each occurrence of k is independently 1-5;
• wherein each occurrence of RG, RH, RJ or RK is independently hydrogen, a linear or branched, substituted or unsubstituted, cyclic or acyclic alkyl moiety, or a substituted or unsubstituted aryl moiety;
• wherein the crosslinker is a moiety derived from a crosslinking reagent capable of conjugating a surface amine of the carrier with a terminal thiol of the linker;
• wherein the carrier is a protein or lipid;
• wherein n is 1, 2, 3 or 4;
• wherein q is 0 or 1;
• wherein each occurrence of RA, RB and RC is independently H or methyl;
• and
• wherein each occurrence of RD, RE and RF independently comprises a carbohydrate domain having the structure:
• wherein a, b, c, d, e, f, g, h, i, x, y and z are each independently 0, 1, 2 or 3, with the proviso that RD, RE, and RF are carbohydrates independently comprised of furanose or pyranose moieties, whereby the sum of b and c is 1 or 2, the sum of d and f is 1 or 2, and the sum of g and i is 1 or 2, and with the proviso that x, y and z are not simultaneously 0;
• wherein R0 is a hydrogen, linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of R1, R2, R3, R4, R5, R6, R7, R8, and R9 is independently hydrogen, OH, ORi, NH2, NHCORi, F, CH2OH, CH2ORi, a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono-, di- or tri)acyloxyalkyl, arylalkyl, or aryl group;
• wherein each occurrence of R1 is independently hydrogen, CHO, CO2Rii, a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl, or aryl group, or a saccharide moiety having the structure:
• wherein Y and Z are independently NH or O;
• wherein k, l, r, s, t, u, v and w are each independently 0, 1 or 2, with the proviso that the v and w bracketed structures represent furanose or pyranose moieties and the sum of l and k is 1 or 2, and the sum of s and u is 1 or 2, and with the proviso that v and w are not simultaneously 0;
• wherein R′0 is hydrogen, a linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of R10, R11, R12, R13, R14 and R15 is independently hydrogen, OH, ORiii, NH2, NHCORiii, F, CH2OH, CH2ORiii, or a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono- di- or tri- )acyloxyalkyl, arylalkyl or aryl group;
• wherein R16 is hydrogen, CO2H, CO2Rii, CONHRii, a substituted or unsubstituted linear or branched chain alkyl or aryl group;
• wherein each ocurrence of Riii is independently hydrogen, CHO, CO2Riv, or a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• and wherein each occurrence of Riiiand Riv is independently hydrogen, or a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl or aryl group, with the limitation that each of RD, RE, and RF comprise a carbohydrate domain, or truncated or elongated version thereof, that is present on tumor cells, and with the further limitation that (i) when q is 0, the linker is —NH(CH2)2—O—(CH2)2NH—, and the carrier is KLH or human serum albumin, then RD, REand RF are not simultaneously STn, and (ii) when q is 0, the linker is —NH(CH2)3(C═O)—, and the carrier is ovine serum albumin, then RD, REand RF are not simultaneously Tn.

A method of inducing antibodies in a subject, wherein the antibodies are capable of specifically binding with human tumor cells, which comprises administering to the subject an amount of a glycoconjugate effective to induce the antibodies, which glycoconjugate has the structure:

• wherein m′, n′ and p′ are integers between about 8 and 20;
• wherein j is an integer between 1 and about 8;
• wherein RV, RA, RB and RC are independently hydrogen, substituted or unsubstituted linear or branched chain lower alkyl or substituted or unsubstituted phenyl;
• wherein RD, RE and RF are independently a carbohydrate domain having the structure:
• wherein a, b, c, d, e, f, g, h, i, x, y and z are each independently 0, 1, 2 or 3, with the proviso that RD, RE, and RF are carbohydrates independently comprised of furanose or pyranose moieties, whereby the sum of b and c is 1 or 2, the sum of d and f is 1 or 2, and the sum of g and i is 1 or 2, and with the proviso that x, y and z are not simultaneously 0;
• wherein R0 is a hydrogen, linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of R1, R2, R3, R4, R5, R6, R7, R8, and R9 is independently hydrogen, OH, ORE, NH2, NHCORi, F, CH2OH, CH2ORi, a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono-, di- or tri)acyloxyalkyl, arylalkyl, or aryl group;
• wherein each occurrence of Ri is independently hydrogen, CHO, CO2Rii a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl, or aryl group, or a saccharide moiety having the structure:
• wherein Y and Z are independently NH or O;
• wherein k, l, r, s, t, u, v and w are each independently 0, 1 or 2, with the proviso that the v and w bracketed structures represent furanose or pyranose moieties and the sum of l and k is 1 or 2, and the sum of s and u is 1 or 2, and with the proviso that v and w are not simultaneously 0;
• wherein R′0 is hydrogen, a linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of R10, R11, R12, R13, R14 and R15 is independently hydrogen, OH, ORiii, NH2, NHCORiii, F, CH2OH, CH2ORiii, or a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono- di- or tri-)acyloxyalkyl, arylalkyl or aryl group;
• wherein R16 is hydrogen, CO2H, CO2Rii, CONHRii, a substituted or unsubstituted linear or branched chain alkyl or aryl group;
• wherein each ocurrence of Riii is independently hydrogen, CHO, CO2Riii, or a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• and wherein each occurrence of Rii and Riv is independently hydrogen, or a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• with the limitation that each of RD, RE, and RF comprise a carbohydrate domain, or truncated or elongated version thereof, that is present on tumor cells.

A method of inducing antibodies in a subject, wherein the antibodies are capable of specifically binding with human tumor cells, which comprises administering to the subject an amount of a glycoconjugate effective to induce antibodies which glycoconjugate has the structure:

• wherein each occurrence of RA, RB and RC is independently H or methyl;
• n is 1, 2, 3 or 4;
• j is 1-8;
• t′ is 1-8;
• S is 0 or 1, wherein when s′=0, the carrier is a lipid, and when s′=1, the carrier is a protein;
• each occurrence of RD, RE and RF is independently a carbohydrate domain selected from the group consisting of Tn, TF, 2,6-STF, 2,6-STn, 3-Ley, 6-Ley, 3,6-STn, 2,3-ST, a carbohydrate having the structure:
• a carbohydrate having the structure:
• a carbohydrate having the structure:
• wherein j, k′ and l′ are each independently 0, 1 or 2;
• and a Ley hexasaccharide having the structure:
• wherein j′ and k′ are each independently 0, 1 or 2.

A method of preventing recurrence of epithelial cancer in a subject which comprises administering an amount of a glycoconjugate effective to induce antibodies against an epithelial cancer cell antigen, which glycoconjugate has the structure:

• wherein the linker is —O—, —NRG—, —NRG(CRHRJ)kNRK—, NRC(CRHRJ)kNRK(C═O)(CRHRJ)kS—, —(CRHRJ)kNRK—, —O(CRHRJ)kNRK—, an oligoester fragment comprising from 2 to about 20 hydroxy acyl residues, a peptidic fragment comprising from 2 to about 20 amino acyl residues, or a linear or branched chain alkyl or aryl carboxylic ester, wherein each occurrence of k is independently 1-5;
• wherein each occurrence of RG, RH, RJ or RKis independently hydrogen, a linear or branched, substituted or unsubstituted, cyclic or acyclic alkyl moiety, or a substituted or unsubstituted aryl moiety;
• wherein the crosslinker is a moiety derived from a crosslinking reagent capable of conjugating a surface amine of the carrier with a terminal thiol of the linker;
• wherein the carrier is a protein or lipid;
• wherein n is 1, 2, 3 or 4;
• wherein q is 0 or 1;
• wherein each occurrence of RA, RB and RC is independently H or methyl;
• and
• wherein each occurrence of RD, RE and RF independently comprises a carbohydrate domain having the structure:
• wherein a, b, c, d, e, f, g, h, i, x, y and z are each independently 0, 1, 2 or 3, with the proviso that RD, RE, and RF are carbohydrates independently comprised of furanose or pyranose moieties, whereby the sum of b and c is 1 or 2, the sum of d and f is 1 or 2, and the sum of g and i is 1 or 2. and with the proviso that x, y and z are not simultaneously 0;
• wherein R0 is a hydrogen, linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of R1, R2, R3, R4, R5, R6, R7, R8, and R9 is independently hydrogen, OH, ORi, NH2, NHCORi, F, CH2OH, CH2ORi, a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono-, di- or tri)acyloxyalkyl, arylalkyl, or aryl group;
• wherein each occurrence of R′ is independently hydrogen, CHO, CO2Rii, a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl, or aryl group, or a saccharide moiety having the structure:
• wherein Y and Z are independently NH or O;
• wherein k, l, r, s, t, u, v and w are each independently 0, 1 or 2, with the proviso that the v and w bracketed structures represent furanose or pyranose moieties and the sum of l and k is 1 or 2, and the sum of s and u is 1 or 2, and with the proviso that v and w are not simultaneously 0;
• wherein R′0 is hydrogen, a linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of R10, R11, R12, R13, R14 and R15 is independently hydrogen, OH, ORiii, NH2, NHCORiii, F, CH2OH, CH2ORiii, or a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono- di- or tri)acyloxyalkyl, arylalkyl or aryl group;
• wherein R16 is hydrogen, CO2H, CO2Rii, CONHRii, a substituted or unsubstituted linear or branched chain alkyl or aryl group;
• wherein each ocurrence of Riii is independently hydrogen, CHO, CO2Riv, or a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• and wherein each occurrence of Rii and Riv is independently hydrogen, or a substituted or unsubstituted linear or branched chain alkyl acyl, arylalkyl or aryl group,
• with the, limitation that each of RD, RE, and RF comprise a carbohydrate domain, or truncated or elongated version thereof, that is present on tumor cells, and with the further limitation that (i) when q is 0, the linker is —NH(CH2)2—O—(CH2)2NH—, and the carrier is KLH or human serum albumin, then RD, RE and RF are not simultaneously STn, and (ii) when q is 0, the linker is —NH(CH2)3(C═O)—, and the carrier is ovine serum albumin, then RD, RE and RF are not simultaneously Tn.

A method of preventing recurrence of epithelial cancer in a subject which comprises administering to a subject an amount of a glycoconjugate effective to induce antibodies, which glycoconjugate has the structure:

• wherein m′, n′ and p′ are integers between about 8 and 20;
• wherein j is an integer between 1 and about 8;
• wherein RV, RA, RB and RC are independently hydrogen, substituted or unsubstituted linear or branched chain lower alkyl or substituted or unsubstituted phenyl;
• wherein RD, RE and RF are independently a carbohydrate domain having the structure:
• wherein a, b, c, d, e, f, g, h, i, x, y and z are each independently 0, 1, 2 or 3, with the proviso that RD, RE, and RF are carbohydrates independently comprised of furanose or pyranose moieties, whereby the sum of b and c is 1 or 2, the sum of d and f is 1 or 2, and the sum of g and i is 1 or 2, and with the proviso that x, y and z are not simultaneously 0;
• wherein R0 is a hydrogen, linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of RI, R2, R3, R4, R5, R6, R7, R8, and R9 is independently hydrogen, OH, ORi, NH2, NHCORi, F, CH2OH, CH2ORi, a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono-, di- or tri)acyloxyalkyl, arylalkyl, or aryl group;
• wherein each occurrence of Ri is independently hydrogen, CHO, CO2Rii, a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl, or aryl group, or a saccharide moiety having the structure:
• wherein Y and Z are independently NH or O;
• wherein k, l, r, s, t, u, v and w are each independently 0, 1 or 2, with the proviso that the v and w bracketed structures represent furanose or pyranose moieties and the sum of l and k is 1 or 2, and the sum of s and u is 1 or 2, and with the proviso that v and w are not simultaneously 0;
• wherein R′0 is hydrogen, a linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• wherein each occurrence of R10, R11, R12, R13, R14 and R15 is independently hydrogen, OH, ORiii, NH2, NHCORiii, F, CH2OH, CH2ORiii, or a substituted or unsubstituted linear or branched chain alkyl, (mono-, di- or tri)hydroxyalkyl, (mono- di- or tri)acyloxyalkyl, arylalkyl or aryl group;
• wherein R16 is hydrogen, CO2H, CO2Riii, CONHRii, a substituted or unsubstituted linear or branched chain alkyl or aryl group;
• wherein each ocurrence of Riii is independently hydrogen, CHO, CO2Riv, or a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• and wherein each occurrence of Rii and Riv is independently hydrogen, or a substituted or unsubstituted linear or branched chain alkyl, acyl, arylalkyl or aryl group;
• with the limitation that each of RD, RE, and RF comprise a carbohydrate domain, or truncated or elongated version thereof, that is present on tumor cells.

A method of preventing recurrence of epithelial cancer in a subject which comprises administering an amount of a glycoconjugate effective to induce antibodies against an epithelial cancer cell antigen, which glycoconjugate has the structure:

• wherein each occurrence of RA, RB and RC is independently H or methyl;
• n is 1 ,2, 3 or 4;
• j is 1-8;
• t′ is 1-8;
• s′is 0 or 1, wherein when s′=0, the carrier is a lipid, and when s′=1, the carrier is a protein;
• each occurrence of RD, REand RFis independently a carbohydrate domain selected from the group consisting of Tn, TF, 2,6-STF, 2,6-STn, 3-Ley, 6-Ley, 3,6-STn, 2,3-ST, a carbohydrate having the structure:
• a carbohydrate having the structure:
• a carbohydrate having the structure:
• wherein j′, k′ and l′ are each independently 0, 1 or 2;
• and a Ley hexasaccharide having the structure:
• wherein j′ and k′ are each independently 0, 1 or 2.

A method of treating cancer in a subject suffering therefrom comprising administering to the subject a therapeutically effective amount of a glycoconjugate having the structure:

• wherein j is 1-8;
• t′ is 1-8;
• s′is 0 or 1, wherein when s′=0, the carrier is a lipid and when s′=1, the carrier is a protein;
• RA is hydrogen or methyl;
• and
• RD is selected from the group consisting of Tn, TF, 2,6-STF, 2,6-STn, 3-Ley, 6-Ley, 3,6-STn, 2,3-ST, a carbohydrate having the structure:
• a carbohydrate having the structure:
• a carbohydrate having the structure:
• wherein j′, k′ and l′ are each independently 0, 1 or 2;
• and a Ley hexasaccharide having the structure:
• wherein j′ and k′ are each independently 0, 1 or 2.

A method of inducing antibodies in a subject, wherein the antibodies are capable of specifically binding with human tumor cells, which comprises administering to the subject an amount of a glycoconjugate effective to induce antibodies which glycoconjugate has the structure:

• wherein j is 1-8;
• t′ is 1-8;
• s′is 0 or 1, wherein when s′0, the carrier is a lipid and when s′=1, the carrier is a protein;
• RA is hydrogen or methyl;
• and
• RD is selected from the group consisting of Tn, TF, 2,6-STF, 2,6-STn, 3-Ley, 6-Ley, 3,6-STn, 2,3-ST, a carbohydrate having the structure:
• a carbohydrate having the structure:
• a carbohydrate having the structure:
• wherein j′, k′ and l′ are each independently 0, 1 or 2;
• and a Ley hexasaccharide having the structure:
• wherein j and k′ are each independently 0, 1 or 2.