Tie complex binding proteins

Abstract

Tie1 and Tie2 are receptor tyrosine kinase proteins that include a transmembrane domain. Tie1 and Tie2 are present on endothelial cells. This disclosure describes agents, such as antibodies, that bind to Tie1, Tie2, and Ang, including ones that inhibit endothelial cell activity.

1 . An isolated protein comprising a heavy chain immunoglobulin variable domain sequence and a light chain immunoglobulin variable domain sequence, wherein the protein binds to Tie1 ectodomain and
 (A) the heavy chain immunoglobulin variable domain sequence comprises one or more of the following properties:   i) a HC CDR1 that includes an amino acid sequence as follows:   (AGSR)-Y-(GVK)-M-(GSVF), (SEQ ID NO:117),   (AGSIMRH)-Y-(GVMK)-M-(GSVMFH) (SEQ ID NO:118), or   (AGSIMRNH)-Y-(AGTVMKPQ)-M-(AGSTVMYWFKH) (SEQ ID NO:119);   ii) a HC CDR2 that includes an amino acid sequence as follows:   X-I-Y-P-S-G-G-X-T-X-Y-A-D-S-V-K-G (SEQ ID NO:120), wherein X is any amino acid,   (GSV)-I-(SY)-P-S-G-G-(WQ)-T-(GY) (SEQ ID NO:121),   (GSV)-I-(SY)-P-S-G-G-(WNQ)-T-(GY) (SEQ ID NO:160),   (GSV)-I-(SY)-P-S-G-G-(WQ)-T-(GY)-Y-A-D-S-V-K-G (SEQ ID NO:122),   (GSVW)-I-(SY)-P-S-G-G-(AGVMYWPQH)-T-(AGSTLVMYFKH) (SEQ ID NO:123), or   X-I-Y-P-S-G-G-(WPS)-T-(YVH)-Y-A-D (SEQ ID NO:704), wherein X is any amino acid;   iii) a HC CDR3 that includes an amino acid sequence as follows:   V-(four or five residues)-F-D-(I/Y) (SEQ ID NO:124),   G-Y-G-P-I-A-P-G-L-D-Y (SEQ ID NO:125),   (GV)-N-Y-Y-(GYD)-S-(SD)-G-Y-G-P-I-A-P-G-L-D-Y (SEQ ID NO:126),   (GVD)-(AGLN)-(LYR)-(GSTLYH)-(GYD)-(AGSYFP)-(SFD)-(AGYD)-(IY)-(GFD)-(YDP)-(IP)-A-P-G-L-D-Y (SEQ ID NO:127),   VNYYDSSGYGPIAPGLDY (SEQ ID NO:128), or   G-X-X-G-(AY)-F-D-(YI) (SEQ ID NO:705), wherein X is any amino acid;   B) and/or the light chain immunoglobulin variable domain sequence comprises one or more of the following properties   i) a LC CDR1 that includes an amino acid sequence as follows:   R-A-S-Q-S-(IV)-S-(SR)-X1-Y-L-(AN) (SEQ ID NO:129),   R-A-S-Q-S-(IV)-S-S-(YS)-L-(ALN) (SEQ ID NO:706),   T-G-T-(SN)-S-D-V-G-(GS)-Y (SEQ ID NO:707),   (SGQ)-(GS)-(DS)-(NS)-(IL)-(GR)-S-(YKN)-(YS)-(VA) (SEQ ID NO:708),   R-A-S-Q-S-V-S-S-X-L (SEQ ID NO:130),   R-A-S-Q-S-(IV)-S-(SR)-(SY)-(LY)-(ALN) (SEQ ID NO:131),   S-X-(ND)-(IV)-(AG)-X1-X2-X3 (SEQ ID NO:142),   T-(GR)-(ST)-S-X5-(ND)-(IV)-(AG)-X1-X2-X3-Y-X4-S (SEQ ID NO:143),   
       wherein X1 is any amino acid (e.g., G or R), X2 is any amino acid (e.g., Y or N), X3 is any amino acid (e.g., F, N, or K), X4 is any amino acid (e.g., aliphatic, e.g., V or A), or
 R-A-S-(REQ)-(GSTRN)-(IV)-(GSTIRN)-(STIRH)-X1-(SYWNH)-(LV)-(ASN) (SEQ ID NO:132), wherein X1 can be serine or absent; 
 ii) a LC CDR2 that includes an amino acid sequence as follows: 
 X-A-S-X-R-A-T (SEQ ID NO:133), wherein X can be any amino acid, 
 (AGD)-A-S-(STN)-R-A-T (SEQ ID NO:134), 
 (DG)-(AV)-S-N-(RL)-(AP)-ST) (SEQ ID NO:709), 
 (AGD)-A-S-(STN)-(LR)-(AEQ)-(ST) (SEQ ID NO:135), 
 (DE)-V-N-N-R-P-S (SEQ ID NO:144), 
 (DE)-(VD)-(STDN)-(YRDN)-R-P-S (SEQ ID NO:145), or 
 (AGTKDEH)-A-S-(STN)-(LR)-(AVEQ)-(ST) (SEQ ID NO:136); and 
 iii) a LC CDR3 that includes an amino acid sequence as follows: 
 Q-Q-(SYFR)-(GSYN)-S-(STYW)-(RP)-(LWR)-(TIY)-T (SEQ ID NO:137), 
 Q-Q-(SYFR)-(GSYN)-S-(STYW)-(RP)-(LWRH)-(TIY) (SEQ ID NO:161), 
 (LQ)-Q-(SYFR)-(GSYN)-(SKN)-(STYW)-(RP)-(LWR)-(TIY)-T (SEQ ID NO:138), 
 Q-Q-X-S-(SN)-(WS)-P-X-T-F (SEQ ID NO:710), wherein X is any amino acid, 
 Y-(TG)-(SG)-S-(PGS)-(TN)-X-(VT) (SEQ ID NO:711), wherein X is any amino acid, 
 Q-Q-(YR)-(GS)-S-(SW)-P-R-X1-T (SEQ ID NO:139), wherein X1 is any amino acid or absent, 
 (LQ)-(LQ)-(SYFRD)-(GSYN)-(STRKN)-(STYWF)-(RP)-(ILMWRH)-(TIY)-(TI) (SEQ ID NO:140), 
 (SQ)-S-(SY)-(ASID)-(GSR)-(ST)-(STRN)-(STYR)-(ATLY)-(SVWQ) (SEQ ID NO:146), or 
 (LQ)-(LRQ)-(SYFRD)-(GSYN)-(ASTRKN)-(STYWF)-(SVRP)-(STILMWRH)-(TIY)-(STI) (SEQ ID NO:141). 
 
     
     
         2 . The protein of  claim 1  wherein the amino acid sequences of the HC variable domain sequence comprises CDR1, CDR2, and CDR3 sequences from the E3 or E3b clone, and the LC variable domain sequence comprises CDR1, CDR2, and CDR3 sequences from the E3 or E3b clone. 
     
     
         3 . The isolated protein of  claim 1  that comprises (i) the HC and/or LC immunoglobulin variable domains of the E3 antibody , (ii) HC and/or LC immunoglobulin variable domain sequences that are at least 85% identical to the HC and LC immunoglobulin variable domains of the E3 or E3b antibody, respectively, or (iii) HC and/or LC immunoglobulin variable domain sequences that are encoded by a nucleic acid that hybridizes with high stringency to a nucleic acid encoding a HC or LC variable domain of E3 or E3b, respectively. 
     
     
         4 . The protein of  claim 1  that inhibits tube formation by HUVEC cells in vitro. 
     
     
         5 . An isolated protein comprising a heavy chain immunoglobulin variable domain sequence and a light chain immunoglobulin variable domain sequence, wherein the protein binds to a Tie1 ectodomain and competes with E3, E3b, G2, p-A1, p-A10, p-B1, p-B3, p-C6, p-D12, p-F3, p-F4, p-G3, s-A10, s-H1, s-A2, s-B2, s-B9, s-C10, s-C2, s-C7, s-D11, s-E1 1, s-G10, or s-H4 for binding to Tie1 or binds to an epitope that overlaps an epitope that is recognized by E3 or that has at least one, two or three residues in common with an epitope that is recognized by E3. 
     
     
         6 . A pharmaceutical composition comprising a protein  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         7 . A method of modulating angiogenesis in a subject, the method comprising administering to a subject an agent that inhibits a pro-angiogenic Tie1 activity or antagonizes an association between Tie1 and Tie2. 
     
     
         8 . The method of  claim 7 , wherein the agent comprises an antibody that binds to the extracellular domain of human Tie1. 
     
     
         9 . The method of  claim 8 , wherein the antibody comprises a protein that binds to a Tie1 ectodomain and comprises a heavy chain (HC) immunoglobulin variable domain sequence and a light chain (LC) immunoglobulin variable domain sequence, the protein further comprising one or more of the following properties:
 (1) at least one of the variable domain sequences comprising at least two CDR of the E3 antibody;   (2) at least one of the variable domain sequences comprising CDR sequences at. least 85% identical, in sum, to the CDRs of the corresponding variable domain of the E3 antibody,   (3) at least one of the variable domains is at least 85% identical to the corresponding immunoglobulin variable domains of the E3 antibody,   (4) the protein competes with E3 for binding to Tie1 or binds to an epitope that overlaps the epitope bound by E3 on Tie1, and   (5) the protein comprises a domain that is encoded by a nucleic acid that hybridizes with high stringency to a nucleic acid encoding a HC or LC variable domain of E3 or E3b.   
     
     
         10 . The method of  claim 9 , wherein the antibody comprises the variable domains of E3 or E3b. 
     
     
         11 . The method of  claim 7 , wherein the agent comprises an antibody that binds to the extracellular domain of human Tie2. 
     
     
         12 . The method of  claim 7 , wherein the agent is administered in an amount and/or a time effective to decrease angiogenesis in a subject. 
     
     
         13 . The method of  claim 7 , wherein the subject has an angiogenesis-dependent cancer or tumor. 
     
     
         14 . The method of  claim 7 , wherein the subject has an inflammatory disorder. 
     
     
         15 . A method of treating an angiogenesis-related disorder, the method comprising:
 providing a first therapy that comprises administering an agent that inhibits a pro-angiogenic acitivity of Tie1 or decreases Tie1-Tie2 interaction in combination with a second therapy.   
     
     
         16 . The method of  claim 15 , wherein the second therapy is an anti-cancer therapy and the subject is diagnosed with cancer. 
     
     
         17 . The method of  claim 16 , wherein the anti-cancer therapy comprises an agent that antagonizes signaling through a VEGF pathway. 
     
     
         18 . A method for detecting the presence of a Tie1 protein, in a sample, in vitro, the method comprising:
 (i) contacting the sample with an Tie1-binding protein according to  claim 1 , under conditions that allow interaction of the Tie1-binding protein and the Tie1 protein to occur; and   (ii) detecting formation of a complex between the Tie1-binding protein and the sample.   
     
     
         19 . A method for detecting and/or localization Tie1 in vivo, the method comprising:
 (i) administering to a human subject a Tie1-binding protein, under conditions that allow interaction of the Tie1-binding protein and the Tie1 protein to occur; and   (ii) detecting formation of a complex between the Tie1-binding protein and a Tie1 molecule of the subject or detecting distribution of the Tie1-binding protein at least one location in the subject.   
     
     
         20 . The method of  claim 19  wherein the Tie1-binding protein is a Tie1 antibody.