Multivalent glycopeptides that tightly bind to target proteins

Patent metadata

Field	Value
Publication number	US-2016304628-A1
Application number	US-201415101221-A
Country	US
Kind code	A1
Filing date	Dec 2, 2014
Priority date	Dec 2, 2013
Publication date	Oct 20, 2016
Grant date	—

Abstract

The invention relates to a glycopolypeptide that includes one or more modified amino acid residues having a sidechain comprising a monosaccharide or an oligosaccharide, wherein the glycopolypeptide binds specifically to a carbohydrate-binding monoclonal antibody with an affinity of less than 100 nM. Immunogenic conjugates that include the glycopolypeptide, and pharmaceutical compositions that include the glycopolypeptide or the immunogenic conjugate are also disclosed. Various method of using the glycopolypeptides, immunogenic conjugates, and pharmaceutical compositions are disclosed, including inducing an immune response, inhibiting viral or bacterial infection, treating a cancerous condition, and detecting a neutralizing antibody.

First claim

1 . A glycopolypeptide comprising one or more modified amino acid residues having a sidechain comprising a monosaccharide or an oligosaccharide, wherein the glycopolypeptide binds specifically to a carbohydrate-binding monoclonal antibody with an affinity of less than 100 nM.

2 - 4 . (canceled)

5 . The glycopolypeptide according to claim 1 , wherein the glycopolypeptide comprises about 10 to about 80 amino acids.

6 . (canceled)

7 . The glycopolypeptide according to claim 1 , wherein the glycopolypeptide comprises from 2 to 5 of said modified amino acid residues.

8 - 12 . (canceled)

13 . The glycopolypeptide according to claim 1 , wherein the modified amino acid residues comprise a sidechain containing a branched or unbranched oligosaccharide that comprises at least about 3 saccharide moieties.

14 - 16 . (canceled)

17 . The glycopolypeptide according to claim 1 , wherein modified amino acid comprises a linker molecule between the polypeptide chain and the monosaccharide or oligosaccharide, the linker molecule comprising

wherein each of R 1 and R 2 is optionally a direct link or independently selected from the group consisting of a linear or branched C 1 to C 18 hydrocarbon that is saturated or mono- or poly-unsaturated, optionally interrupted by one or more non-adjacent —O—, —C(═O)—, or —NR 4 —; a substituted or unsubstituted C 3 to C 10 cycloalkanediyl, a substituted or unsubstituted aryl diradical; a substituted or unsubstituted heteroaryl diradical; a monosaccharide diradical; or a disaccharide diradical;
R 3 is optional and can be —O—, —S—, or —NR 4 —; and
R 4 iS H or a C 1 to C 10 alkyl.

18 . (canceled)

19 . The glycopolypeptide according to claim 1 , wherein the glycopolypeptide binds specifically to the carbohydrate-binding monoclonal antibody with an affinity that is substantially the same as or lower than the affinity of the carbohydrate-binding monoclonal antibody to its naturally occurring binding partner.

20 . The glycopolypeptide according to claim 1 , wherein the carbohydrate-binding monoclonal antibody is neutralizing against a pathogen.

21 - 22 . (canceled)

23 . The glycopolypeptide according to claim 1 , wherein the carbohydrate-binding monoclonal antibody is cytotoxic against a cancer cell.

24 - 25 . (canceled)

26 . The glycopolypeptide according to claim 20 , wherein the carbohydrate-binding, neutralizing monoclonal antibody is 2G12 and the glycopolypeptide comprises the sequence X XSIPXYTY (SEQ ID NO: 2) where X is optional and can be any amino acid and X is the modified amino acid residue to which the oligosaccharide is linked.

27 . The glycopolypeptide according to claim 26 , wherein said glycopolypeptide comprises the sequence:

Sequence
SEQ ID NO:

X DTLHLKQIGG X PNCITQQDVR X TSIPYTYTWP
 3

X LLK X VDQSRL X PVPGIGVTLH X RSIPYSYLPI
 4

X RSTLNSLEYR X QYATEDPRIR X ASIPYTYWWP
 5

ATKTNCKREKT X DNHVTI X RSIPWYTYRWLPN
 6

X ATKTNFKREKT X DNHVTI X RSIPWYTYRWLPN
 7

X ATRTNCKREKT X DNHVTI X RSIPWYTYRWLPN
 8

X ATKTSCKREKT X DNHVTI X RSIPWYTYRWLPN
 9

X VLPTIISTNVNPFR X LSIPTYTYL X PITWGEI
10

X TSIPYTYLNRSLWTNYRVNSWS X SKNVNV X PL
11

X ERPSL X CGLS X LTSGGTQSSV X RSIPFYTYWW
12

X ATKTN KREKT X DNHVTI X RSIPWYTYRWLPN
53

X ATKTN KREKT X DNHVTI X RSIPWYTYRWLPN
54

X RSIPWYTYRWLPN
55

X DTLHLKQIGG X PN ITQQDVR X TSIPYTYTWP
62

28 . The glycopolypeptide according to claim 20 , wherein the carbohydrate-binding, neutralizing monoclonal antibody is 2G12 and the glycopolypeptide comprises the sequence:

Sequence
SEQ ID NO:

X HPYNTSRTSA XX AALK X QVTD X YALALFHRIL
13

X SPHLPVLLCK X VLNDGRRIVQ X SCELP X VRRS
14

X L X FIRIYPTR X QYVYHAPLLT X VR X SPTGPLI
15

X CYVTVIPA X N X PEARLGIVCH X PGIRRGKALY
16

X SPHLPVLL K X VLNDGRRIVQ X S ELP X VRRS
52

XX AALK X QVTD X YALALFHRIL
56

wherein X is the modified amino residue to which the oligosaccharide is linked.

29 . A glycopolypeptide comprising from three to five modified amino acid residues having a sidechain comprising a branched oligosaccharide containing 9 mannose moieties, wherein the glyco

Assignees

• Univ Brandeis

Inventors

• Krauss Isaac J
• Horiya Satoru

Classifications

• C07K16/1145

  Env proteins, e.g. gp41, gp110/120, gp160, V3, principal neutralising domain [PND] or CD4-binding site · CPC title

• G01N2405/00

  Assays, e.g. immunoassays or enzyme assays, involving lipids · CPC title

• G01N33/6845

  Methods of identifying protein-protein interactions in protein mixtures · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• C12N2740/16134

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title