Process for the modification of a glycoprotein using a glycosyltransferase that is or is derived from a ß(1,4)-n-acetylgalactosaminyltransferase

A glycoprotein comprising a glycan according to formula (4) or (5):

• wherein:
• b is 0 or 1;
• d is 0 or 1;
• e is 0 or 1;
• G is a monosaccharide, or a linear or branched oligosaccharide comprising 2 to 20 sugar moieties;
• and
• Su(A) is according to formula (6):
• wherein:
• a is 0 or 1;
• f is 0 or 1;
• g is 0 or 1;
• U is [C(R1)2]n or [C(R1)2]p—O—[C(R1)2C(R1)2O]o—[C(R1)2]q, wherein n is an integer in the range of 1 to 24;
• o is an integer in the range of 0 to 12;
• p and q are independently 0, 1 or 2;
• and R1 is independently selected from the group consisting of H, F, Cl, Br, I, OH and an optionally substituted C1-C24 alkyl group;
• T is a C3-C12 (hetero)arylene group, wherein the (hetero)arylene group is optionally substituted;
• A is selected from the group consisting of:
• (a) —N3
• (b) —C(O)R3 wherein R3 is an optionally substituted C1-C24 alkyl group;
• (c) (hetero)cycloalkynyl group or a —(CH2)iC≡C—R4 moiety wherein i is 0-10 and R4 is hydrogen or an optionally substituted C1-C24 alkyl group;
• (d) —SH
• (e) —SC(O)R8
• wherein R8 is an, optionally substituted, C1-C24 alkyl group or phenyl group;
• (f) —SC(V)OR8
• wherein V is O or S, and R8 is an, optionally substituted, C1-C24 alkyl group or phenyl group;
• (g) —X
• wherein X is selected from the group consisting of F, Cl, Br and I;
• (h) —OS(O)2R5
• wherein R5 is selected from the group consisting of C1-C24 alkyl groups, C6-C24 aryl groups, C7-C24 alkylaryl groups and C7-C24 arylalkyl groups, the alkyl groups, aryl groups, alkylaryl groups and arylalkyl groups being optionally substituted;
• (i) R12
• wherein R12 is selected from the group consisting of, optionally substituted, terminal C2-C24 alkenyl groups, C3-C5 cycloalkenyl groups and C4-C8 alkadienyl groups;
• and
• (j) R13
• wherein R13 is an optionally substituted terminal C3-C24 allenyl group;
• and
• (k) N(R17)2
• wherein R17 is independently selected from the group consisting of H and C1-C12 alkyl groups;
• Z is CH2, CF2 or C(O);
• or Z is CHOH with the proviso that when Z is CHOH, g is 0, f is 1 and a is 0 or 1;
• Y is selected from the group consisting of O, S, N(R15), N(R15)C(O), N(R15)C(O)N(R15), N(R15)C(O)O, OC(O)N(R15)S(O)2N(R15) and N(R15)C(O)N(R15)S(O)2O, wherein R15 is independently selected from the group consisting of H, C1-C12 alkyl groups and (U)f-(T)a-A wherein f, a, U, T and A are as defined above;
• and
• R14 is selected from the group consisting of:
• wherein:
• a, f, T, A and U are as defined above;
• h is 0 or 1;
• and
• W is selected from the group consisting of O, S, NR15, NHS(O)2O and NHS(O)2NR15,
• wherein R15 is as defined above.

The bioconjugate according to formula (75), (76) or (77):

• wherein:
• Pr is a protein
• y is independently an integer in the range of 1 to 24;
• CG is a connecting group that connects Su to Sp or D;
• Sp is a spacer;
• D is a target molecule;
• j is independently 1, 2, 3, 4 or 5;
• k is independently an integer in the range of 1 to 10;
• m is 0 or 1;
• b is 0 or 1;
• d is 0 or 1;
• e is 0 or 1;
• G is a monosaccharide, or a linear or branched oligosaccharide comprising 2 to 20 sugar moieties;
• Su is according to formula (78):
• wherein:
• a is 0 or 1;
• f is 0 or 1;
• g is 0 or 1;
• U is [C(R1)2]n or [C(R1)2]p—O—[C(R1)2C(R1)2O]o—[C(R1)2]q, wherein n is an integer in the range of 1 to 24;
• o is an integer in the range of 0 to 12;
• p and q are independently 0, 1 or 2;
• and R1 is independently selected from the group consisting of H, F, Cl, Br, I, OH and an optionally substituted C1-C24 alkyl group;
• T is a C3-C12 (hetero)arylene group, wherein the (hetero)arylene group is optionally substituted;
• Z is CH2, CF2 or C(O);
• or Z is CHOH with the proviso that g is 0, f is 1 and a is 0 or 1;
• Y is selected from the group consisting of O, S, N(R15), N(R15)C(O), N(R15)C(O)N(R15) N(R15)C(O)O, OC(O)N(R15)S(O)2N(R15) and N(R15)C(O)N(R15)S(O)2O, wherein R15 is independently selected from the group consisting of H, C1-C12 alkyl groups and (U)f-(T)a-A wherein f, a, U, T and A are as defined above;
• and
• R14 is selected from the group consisting of:
• wherein:
• a, f, T and U are as defined above;
• h is 0 or 1;
• and
• W is selected from the group consisting of O, S, NR15, NHS(O)2O and NHS(O)2NR15, wherein R15 is as defined above,
• A is selected from the group consisting of:
• (a) —N3
• (b) —C(O)R3
• wherein R3 is an unsubstituted or substituted C1-C24 alkyl group;
• (c) (hetero)cycloalkynyl group or a —(CH2)iC≡C—R4 moiety
• wherein i is 0-10 and R4 is hydrogen or an unsubstituted or substituted C1-C24 alkyl group;
• (d) —SH
• (e) —SC(O)R8
• wherein R8 is an unsubstituted or substituted C1-C24 alkyl group or phenyl group;
• (f) —SC(V)OR8
• wherein V is O or S, and R8 is an unsubstituted or substituted C1-C24 alkyl group or phenyl group;
• (g) —X
• wherein X is selected from the group consisting of F, Cl, Br and I;
• (h) —OS(O)2R5
• wherein R5 is selected from the group consisting of C1-C24 alkyl groups, C6-C24 aryl groups, C7-C24 alkylaryl groups and C7-C24 arylalkyl groups, the alkyl groups, aryl groups, alkylaryl groups and arylalkyl groups being unsubstituted or substituted;
• (i) R12
• wherein R12 is selected from the group consisting of unsubstituted or optionally substituted terminal C2-C24 alkenyl groups, C3-C5cycloalkenyl groups and C4-C8 alkadienyl groups;
• and
• (j) R13
• wherein R13 is an unsubstituted or substituted terminal C3-C24 allenyl group;
• and
• (k) N(R17)2
• wherein R17 is independently selected from the group consisting of H and C1-C12 alkyl groups;
• and Su is connected via C1 to C4 of the GlcNAc moiety via a β-1,4-O-glycosidic bond and to CG via Z, Y, U or T.