US 2017/0130256 A1 · PROCESS FOR THE MODIFICATION OF A GLYCOPROTEIN USING A GLYCOSYLTRANSFERASE THAT IS OR IS DERIVED FROM A ß(1,4)-N-ACETYLGALACTOSAMINYLTRANSFERASE

Abstract

The present invention relates to a process for the enzymatic modification of a glycoprotein. The process comprises the step of contacting a glycoprotein comprising a glycan comprising a terminal GlcNAc-moiety, in the presence of glycosyltransferase that is, or is derived from, a β-(1,4)-N-acetylgalactosaminyltransferase, with a non-natural sugar-derivative nucleotide. The non-natural sugar-derivative nucleotide is according to formula (3): wherein A is selected from the group consisting of —N3, —C(O)R3, —(CH2)iC≡C—R4, —SH, —SC(O)R8, —SC(0)OR8, —SC(S)OR8, —F, —CI, —Br —I, —OS(O)2R5, terminal C2-C24 alkenyl groups, C3-C5 cycloalkenyl groups, C4-C8 alkadienyl groups, terminal C3-C24 allenyl groups and amino groups. The invention further relates to a glycoprotein obtainable by the process according to the invention, to a bioconjugate that can be obtained by conjugating the glycoprotein with a linker-conjugate, and to β-(1,4)-N-acetylgalactosaminyltransferases that can be used in preparing the the glycoprotein according to the invention.

Claims18

Dependent claims

-23. (canceled)

A bioconjugate having 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;
Su is according to formula (78):
and wherein
b is 0 or 1;
d is 0 or 1;
e is 0 or 1;
and
G is a monosaccharide, or a linear or branched oligosaccharide comprising 2 to 20 sugar moieties;
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, and T are as defined above;
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;
and
R14 is —NHC(O)Me,
and Su is connected via C1 to C4 of the GlcNAc moiety via a b-1,4-O-glycosidic bond and to CG via Z, Y, U or T.

50 more elements

A process for the modification of a glycoprotein, the process comprising contacting a glycoprotein comprising a glycan comprising a terminal GlcNAc moiety, with a sugar-derivative nucleotide Su(A)-Nuc in the presence of a glycosyltransferase, wherein:

(i) the glycosyltransferase is, or is derived from, a β-(1,4)-N-acetylgalactosaminyltransferase;
(ii) the glycan comprising a terminal GlcNAc-moiety is according to formula (1) or (2):
wherein:
b is 0 or 1;
d is 0 or 1;
e is 0 or 1;
and
G is a monosaccharide, or a linear or branched oligosaccharide comprising 2 to 20 sugar moieties;
and
(iii) the sugar-derivative nucleotide Su(A)-Nuc is according to formula (3):
wherein:
a is 0 or 1;
f is 0 or 1;
g is 0 or 1;
Nuc is a nucleotide;
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 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)O and NHS(O)2NR15, wherein R15 is as defined above.

50 more elements

Dependencies are inferred from the claim text.

Facts

Application

15/318,248Filed 25 Apr 2016

Priority

EP 15164864.923 Apr 2015

Published

11 May 2017

Term ends

Not estimatedThe source records don’t support an estimate.

Classification17 codes

C12P 21/005
Preparation of peptides or proteins: glycopeptides, glycoproteins
A61K 47/61
The modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule: the organic macromolecular…
A61K 47/64
The modifying agent being a protein, peptide or polyamino acid: drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying…
A61K 47/6803
Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent: drugs conjugated to an antibody or…

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

Life of this patent

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

Life of this patent

Abstract

Claims18