Tie2-binding agents and methods of use

Patent metadata

Field	Value
Publication number	US-2021332142-A1
Application number	US-202117208292-A
Country	US
Kind code	A1
Filing date	Mar 22, 2021
Priority date	Mar 24, 2020
Publication date	Oct 28, 2021
Grant date	—

Abstract

The invention provides Tie-2 antibodies and fragments thereof and conjugates and methods of using the same.

First claim

What is claimed is:

1 . An isolated antibody or antigen-binding fragment thereof that specifically binds to Tie2, wherein the antibody or fragment thereof comprises:
a heavy chain variable domain (VH) comprising (a) CDR-H1 comprising the amino acid sequence NTDIS (SEQ ID NO:3), (b) CDR-H2 comprising the amino acid sequence RISPSDGNTYYADSVKG (SEQ ID NO:4), and (c) CDR-H3 comprising the amino acid sequence RTRWASX1AX2DY (SEQ ID NO:5), where X1 is M, L, K, F, Y, R, N, Q, H or W and/or or X2 is F, Y L, Q, I, K, or H, and a light chain variable domain (VL) comprising (d) CDR-L1 comprising the amino acid sequence RASQDVSTAVA (SEQ ID NO:8), (e) CDR-L2 comprising the amino acid sequence SASFLYS (SEQ ID NO:9), and (f) CDR-L3 comprising the amino acid sequence QQSYTTPPT (SEQ ID NO:10).

2 . The antibody or antigen-binding fragment thereof according to claim 1 , wherein the CDR-H3 comprises the amino acid sequence RTRWASWAMDY (SEQ ID NO:6).

3 . The antibody or antigen-binding fragment thereof according to claim 1 or 2 , wherein the CDR-H3 comprises the amino acid sequence RTRWASWAFDY (SEQ ID NO:7).

4 . The antibody or antigen-binding fragment thereof of any one of the preceding claims, which is a monoclonal antibody, a humanized antibody, an antibody fragment, and/or a chimeric antibody.

5 . The antibody or antigen-binding fragment thereof of any one of the preceding claims, which is a Fab fragment that binds Tie2.

6 . The antibody or antigen-binding fragment thereof of any one of claims 1 to 5 , comprising a VL domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:21 and a VH domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:20.

7 . The antibody or antigen-binding fragment thereof of any one of claims 1 to 6 , comprising a VL domain comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:21 and a VH domain comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:20.

8 . The antibody or antigen-binding fragment thereof of any one of the preceding claims, wherein the antibody comprises the VL sequence of SEQ ID NO:21 and a VH sequence of SEQ ID NO:20.

9 . The antibody or antigen-binding fragment thereof of any one of claims 1 to 8 , wherein the antibody comprises an engineered cysteine, wherein
the engineered cysteine is selected from a T120C, G166C, G178C, T187C, and T209C in the HC; or
the engineered cysteine is selected from Q124C, R142C, Q155C, L201C, T206C, K107C, K126C, and K149C in the LC;
wherein the residue number of the engineered cysteine is according to EU numbering.

10 . The antibody or antigen-binding fragment thereof of claim 9 , wherein the engineered cysteine is selected from T209C in the HC and T206C in the LC.

11 . The antibody or antigen-binding fragment thereof of claim 9 or 10 , wherein the engineered cysteine is T206C in the LC.

12 . The antibody or antigen-binding fragment thereof of claim 9 or 10 , wherein the engineered cysteine is T209C in the HC.

13 . The antibody or antigen-binding fragment thereof of any one of the preceding claims, comprising a LC comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:25 and a HC comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:55.

14 . The antibody or antigen-binding fragment thereof of any one of the preceding claims, comprising a LC comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:25 and a HC comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:55.

15 . The antibody or antigen-binding fragment thereof of any one of the preceding claims, wherein the antibody comprises a LC comprising the sequence of SEQ ID NO:25 and a HC comprising the sequence of SEQ ID NO:55.

16 . The antibody or antigen-binding fragment thereof of any one of the preceding claims, wherein the HC constant regions terminates at position 221 (EU numbering).

17 . An antibody or antigen-binding fragment thereof that specifically binds to Tie-2 wherein the antibody comprises a HC sequence of SEQ ID NO:55 and a LC sequence of SEQ ID NO:25.

18 . An antibody or antigen-binding fragment thereof that specifically binds to Tie-2 wherein the antibody comprises a HC sequence of SEQ ID NO:23 and a LC sequence of SEQ ID NO:56.

19 . An isolated nucleic acid encoding the antibody or antigen-binding fragment thereof of any of the preceding claims.

20 . A host cell comprising the nucleic acid of claim 19 .

21 . A conjugate that binds to Tie2, wherein the conjugate comprises at least two, at least three, at least four, at least five, at least six, at least seven, or at least eight antibodies or antigen-binding fragment thereof according to any one of claims 1 to 18 , wherein each of the antibodies or antigen-binding fragment thereof is linked to a multimerizing moiety.

22 . The conjugate of claim 21 , wherein the conjugate causes a reduction in Tie2 protein levels in an in vitro assay by less than 25%, less than 50% or less than 75%.

23 . The conjugate of claim 21 or 22 , wherein the conjugate does not cause a reduction in Tie-2 protein levels in an in vitro assay by more than 25%, more than 50%, or more than 75%.

24 . The conjugate of any one of claims 21 to 23 , wherein the multimerizing moiety comprises a polyol, a polypeptide, and/or a peptide.

25 . The conjugate of any one of claims 21 to 24 , wherein the multimerizing moiety comprising a polyol, wherein the polyol is a multi-armed polyol, selected from a dimer, a tetramer, a hexamer, and an octamer.

26 . The conjugate of claim 25 , wherein the multi-armed polyol is a hexamer.

27 . The conjugate of any one of claims 24 to 26 , wherein the polyol is a polyethylene glycol (PEG).

28 . The conjugate of any one of claims 24 to 27 , wherein the polyol is covalently linked to the at least two antibodies or antigen-binding fragment thereof through a free sulfhydryl group of an engineered cysteine amino acid, wherein the engineered cysteine is selected from the group consisting of
T120C, G166C, G178C, T187C, and T209C in the HC; or
the engineered cysteine is selected from the group consisting of Q124C, R142C, Q155C, L201C, T206C, K107C, K126C, and K149C in the LC;
wherein the residue number is according to EU numbering.

29 . The conjugate of claim 28 , wherein the engineered cysteine is T206C in the LC, wherein the residue number is according to EU numbering.

30 . The conjugate of claim 28 , wherein the engineered cysteine is T209C in the HC, wherein the residue number is according to EU numbering.

31 . The conjugate of any one of claims 25 to 27 , wherein the polyol is covalently linked to the at least one antibody through a free amino group of a lysine amino acid.

32 . The conjugate according to any one of claims 27 to 31 , wherein the PEG has a weight average molecular weight of from about 500 Daltons (Da) to about 300,000 Da.

33 . The conjugate acco

Assignees

• Genentech Inc

Inventors

• Yan Minhong
• Zhang Gu
• Agard Nicholas John
• Dicara Danielle Marie
• Hass Philip E
• Hang Julie Q
• Christensen Erin L
• Morse Robert Paul
• Sanowar Sarah
• Shivva Vittal

Classifications

• C07K2317/94

  Stability, e.g. half-life, pH, temperature or enzyme-resistance · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/71

  Decreased effector function due to an Fc-modification · CPC title

• C07K2317/55

  Fab or Fab' · CPC title

• C07K16/2863 Primary

  against receptors for growth factors, growth regulators · CPC title