Anti-TLR4 antibodies and uses thereof

Patent metadata

Field	Value
Publication number	US-10400044-B2
Application number	US-201615276202-A
Country	US
Kind code	B2
Filing date	Sep 26, 2016
Priority date	Mar 29, 2012
Publication date	Sep 3, 2019
Grant date	Sep 3, 2019

Abstract

Disclosed are antibodies that specifically bind Toll-like Receptor 4 (TLR-4), and to methods of using the anti-TLR4 antibodies as therapeutics and diagnostic agents.

First claim

The invention claimed is:

1. A method of blocking or neutralizing Toll-like receptor 4 (TLR4) signaling comprising administering to a subject in need thereof an isolated antibody—that specifically binds TLR4 in an amount sufficient to block or neutralize TLR4 signaling, wherein the antibody comprises a heavy chain with three complementarity determining regions (CDRs) comprising a variable heavy chain complementarity determining region 1 (CDRH1) amino acid sequence of GYSITGGYS (SEQ ID NO: 25); a variable heavy chain complementarity determining region 2 (CDRH2) amino acid sequence of IHYSGYT (SEQ ID NO: 26); and a variable heavy chain complementarity determining region 3 (CDRH3) amino acid sequence of ARKDSG(X 1 )(X 2 )(X 3 )PY (SEQ ID NO: 111), where X 1 is N, Q, D or E, X 2 is any hydrophobic amino acid, and X 3 is any hydrophobic amino acid; and a light chain with three CDRs comprising a variable light chain complementarity determining region 1 (CDRL1) amino acid sequence of QSISDH (SEQ ID NO: 37); a variable light chain complementarity determining region 2 (CDRL2) amino acid sequence of YAS (SEQ ID NO: 38); and a variable light chain complementarity determining region 3 (CDRL3) amino acid sequence of QQGHSFPLT (SEQ ID NO: 39).

2. The method of claim 1 , wherein the antibody further comprises an amino acid substitution in the gamma heavy chain constant region at EU amino acid position 325 and an amino acid substitution at EU amino acid position 328, wherein the amino acid substituted at EU amino acid position 325 is serine, and wherein the amino acid substituted at EU amino acid position 328 is phenylalanine.

3. The method of claim 2 , wherein the subject is a human.

4. The method of claim 3 , wherein the subject has an autoimmune disease or an inflammatory disorder.

5. A method of blocking or neutralizing Toll-like receptor 4 (TLR4) signaling comprising administering to a subject in need thereof an isolated antibody—that specifically binds TLR4 in an amount sufficient to block or neutralize TLR4 signaling, wherein the antibody comprises a heavy chain with three complementarity determining regions (CDRs) comprising a variable heavy chain complementarity determining region 1 (CDRH1) amino acid sequence of G(X 1 )PI(X 2 )(X 3 )GYS (SEQ ID NO: 110), where X 1 is F or Y, X 2 is R, G or W, X 3 is Y, F or G;
a variable heavy chain complementarity determining region 2 (CDRH2) amino acid sequence of IHYSGYT (SEQ ID NO: 26); and a variable heavy chain complementarity determining region 3 (CDRH3) amino acid sequence of ARKDSG(X 1 )(X 2 )(X 3 )PY (SEQ ID NO: 111), where X 1 is N, Q, D or E, X 2 is any hydrophobic amino acid, and X 3 is any hydrophobic amino acid; and a light chain with three CDRs comprising a variable light chain complementarity determining region 1 (CDRL1) amino acid sequence of QSISDH (SEQ ID NO: 37); a variable light chain complementarity determining region 2 (CDRL2) amino acid sequence of YAS (SEQ ID NO: 38); and a variable light chain complementarity determining region 3 (CDRL3) comprising the amino acid sequence of QQG(X 1 )(X 2 )(X 3 )P(X 4 )T (SEQ ID NO: 112), where X 1 is Y or N, X 2 is D or E, X 3 is F or Y, and X 4 is any hydrophobic amino acid, or the amino acid sequence of QQGHSFPLT (SEQ ID NO: 39).

6. The method of claim 5 , wherein the antibody further comprises an amino acid substitution in the gamma heavy chain constant region at EU amino acid position 325 and an amino acid substitution at EU amino acid position 328, wherein the amino acid substituted at EU amino acid position 325 is serine, and wherein the amino acid substituted at EU amino acid position 328 is phenylalanine.

7. The method of claim 6 , wherein the subject is a human.

8. The method of claim 7 , wherein the subject has an autoimmune disease or an inflammatory disorder.

9. A method of blocking or neutralizing Toll-like receptor 4 (TLR4) signaling comprising administering to a subject in need thereof an isolated antibody—that specifically binds TLR4 in an amount sufficient to block or neutralize TLR4 signaling, wherein the antibody comprises a heavy chain with three complementarity determining regions (CDRs) comprising a variable heavy chain complementarity determining region 1 (CDRH1) amino acid sequence of G(X 1 )PI(X 2 )(X 3 )GYS (SEQ ID NO: 110), where X 1 is F or Y, X 2 is R, G or W, X 3 is Y, F or G; a variable heavy chain complementarity determining region 2 (CDRH2) amino acid sequence of IHYSGYT (SEQ ID NO: 26); and a variable heavy chain complementarity determining region 3 (CDRH3) amino acid sequence of ARKDSG(X 1 )(X 2 )(X 3 )PY (SEQ ID NO: 111), where X 1 is N, Q, D or E, X 2 is any hydrophobic amino acid, and X 3 is any hydrophobic amino acid; and a light chain with three CDRs comprising a variable light chain complementarity determining region 1 (CDRL1) amino acid sequence of QSISDH (SEQ ID NO: 37); a variable light chain complementarity determining region 2 (CDRL2) amino acid sequence of YAS (SEQ ID NO: 38); and a variable light chain complementarity determining region 3 (CDRL3) amino acid sequence of QQG(X 1 )(X 2 )(X 3 )P(X 4 )T (SEQ ID NO: 112), where X 1 is Y or N, X 2 is D or E, X 3 is F or Y, and X 4 is any hydrophobic amino acid.

10. The method of claim 9 , wherein the antibody comprises a CDRH1 amino acid sequence of GYSITGGYS (SEQ ID NO: 25); GFPIRYGYS (SEQ ID NO: 55), GYPIRFGYS (SEQ ID NO: 56), GYPIRHGYS (SEQ ID NO: 57), GFPIGQGYS (SEQ ID NO: 58), GYPIWGGYS (SEQ ID NO: 59) or GYPIGGGYS (SEQ ID NO: 60); a CDRH2 amino acid sequence of IHYSGYT (SEQ ID NO: 26); and a CDRH3 amino acid sequence selected from the group consisting of ARKDSGNYFPY (SEQ ID NO: 27); ARKDSGQLFPY (SEQ ID NO: 33); and ARKDSGDYFPY (SEQ ID NO: 35); a CDRL1 amino acid sequence of QSISDH (SEQ ID NO: 37); a CDRL2 amino acid sequence of YAS (SEQ ID NO: 38); and a CDRL3 amino acid sequence of QQGHSFPLT (SEQ ID NO: 39), QQGNDFPVT (SEQ ID NO: 61), QQGYDEPFT (SEQ ID NO: 62), QQGYDFPLT (SEQ ID NO: 63), QQGYDYPLT (SEQ ID NO: 64) or QQGYEFPLT (SEQ ID NO: 65).

11. The method of claim 10 , wherein the antibody further comprises an amino acid substitution in the gamma heavy chain constant region at EU amino acid position 325 and an amino acid substitution at EU amino acid position 328, wherein the amino acid substituted at EU amino acid position 325 is serine, and wherein the amino acid substituted at EU amino acid position 328 is phenylalanine.

12. The method of claim 9 , wherein the antibody comprises a heavy chain variable amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 6, 8, 10, 12, 14, 16, 18, 20, 22, 67, 69, 71, 73, 75 or 77 and comprises a light chain variable amino acid comprising the amino acid sequence of SEQ ID NO: 4, 79, 81, 83, 85 or 87.

13. The method of claim 9 , wherein the antibody comprises a combination of a variable heavy chain amino acid sequence and a variable light chain amino acid sequence selected from the group consisting of:
(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 6 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 4;
(b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 8 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 4;
(c) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 4;
(d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 12 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 4;
(e) a heavy chain variable region compris

Assignees

• Novimmune Sa

Inventors

• Rousseau Francois
• Loyau Jeremy
• Fischer Nicolas
• Elson Greg
• Kosco-Vilbois Marie

Classifications

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P29/00

  Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] · CPC title

• C07K2317/34

  Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues · CPC title

• C07K2317/56

  variable (Fv) region, i.e. VH and/or VL · CPC title