Ang-2 Binding Complexes and Uses Thereof

Abstract

Complexes containing one or more modular recognition domains (MRDs) and MRDs attached to scaffolds including antibodies are described. The manufacture of these complexes are the use of these complexes to treat and diagnose diseases and disorders are also described.

1 . A complex comprising a modular recognition domain (MRD) capable of binding ANG-2, wherein the MRD comprises an amino acid sequence having the formula
 X 1 X 2 X 3 X 4 X 5 X 6 MPMDX 11 X 12 EX 14 X 15 LYEX 19 X 20 X 21 X 22  (SEQ ID NO:125) and wherein:   X 1  is 1-10 amino acid residues;   X 2  is L, A, V, P, 1, W, F, M, S, N, F, G, T, H, Y, or C;   X 3  is N, Q, G, S, T, E, D, Y, M, V, L, or I;   X 4  is N, Q, G, S, T, Y, F, E, P, A, or H;   X 5  is N, Q, O, S, T, Y, E, H, L, A, V, P, I, W, F, or M;   X 6  is V, M, A, F, L, P, I, W, or Y;   X 11  is Q, G, S, T, Y, L, V, P, I, W, F, M, K, R, H;   X 12  is D, E, N, Q, G, S, T, Y, P, W, K, R, or H;   X 14  is A, D, N, G, S, T, Y, V, P, I, W, F, M, K, R, or H;   X 15  is V, M, A, F, L, P, I, W, Y, D, E, T, H, or Norleucine;   X 19  is A, E, D, G, S, T, Y, L, V, P, I, W, F, K, R, or H;   X 20  is E, D, V, M, A, F, L, P, 1, W, Y, K, R, H, or Norleucine;   X 21  is V, M, A, F, L, P, I, W, C, Y, or Norleucine; and   X 22  is 1-10 amino acid residues.   
     
     
         2 . The complex of  claim 1 , wherein said MRD comprises an amino acid sequence having the formula X 1 X 2 X 3 X 4 X 5 X 6 MPMDX 11 X 12 EX 14 X 15 LYEX 19 X 20 X 21 X 22  (SEQ ID NO:131) and wherein:
 X, is 1-10 amino acid residues;   X 2  is A, V, I, or C;   X 3  is D, N, or Q;   X 4  is S, or T;   X 5  is Q, E, or N;   X 6  is I, A, V, P, I, W, F, or M;   X 11  is Q, G, S, T, Y, L, V, P, I, W, F, M, K, R, or H;   X 12  is D, E, S, K, or R;   X 14  is A, D, G, V, P, I, W, F, M, K, R, or H;   X 15  is L, I, or Norleucine;   X 19  is A, E, D, G, S, T, Y, L, V, P, 1, W, F, M, K, R, or H;   X 20  is L, V, Norleucine, or F;   X 21  is L, A, V, I, or Norleucine; and   X 22  is 1-10 amino acid residues.   
     
     
         3 . The complex of  claim 1 , wherein the MRD comprises an amino acid having the formula X 1 AQTNFMPMDX 11 X 12 EX 14 LLYEX 19 X 20 FI (SEQ ID NO:134) wherein:
 X 1  is 1-10 amino acid residues;   X 11  is Q, G, S, T, Y, L, V, P, I, W, F, M, K, R, or H;   X 12  is D, E, N, Q, O, S, T, Y, P, W, K, R, or H;   X 14  is A, D, G, V, P, I, W, F, M, K, T, or H;   X 19  is A, E, D, G, S, T, Y, L, V, P, I, W, F, M, K, R, or H; and   X 20  is E, D, V, M, A, F, L, P, I, W, Y, K, R, H, or Norleucine.   
     
     
         4 . The complex of  claim 1 , wherein the MRD comprises an amino acid having the formula X 1 AQTNFMPMDX 11 X 12 EX 14 LLYEX 19 X 20 FI (SEQ ID NO:139)) wherein:
 X 1  is 1-10 amino acid residues;   X 11  is Q, Y, V, P, W, F, K, or R;   X 12  D, E, N, Q, or G;   X 14  is A, D, N, G, S, T, Y, V, P, W, F, M, K, R, or H;   X 19  is A, E, D, G, S, T, Y, L, V, P, I, W, F, M, K, R, or H; and   X 20  is L, V, Norleucine, or F.   
     
     
         5 . The complex of  claim 1 , wherein the MRD comprises amino acids AQTNFMPM DQEFALLYEEFI (SEQ ID NO:108). 
     
     
         6 . The complex of  claim 1 , wherein the MRD comprises amino acids AQTNFMPM DQDEALLYFEEFI (SEQ ID NO:109). 
     
     
         7 . The complex of  claim 1 , wherein the MRD comprises amino acids AQTNFMPM DQDEALLYEQEQFI (SEQ ID NO 110). 
     
     
         8 . The complex of  claim 1 , wherein the MRD comprises amino acids AQTNFMPM DQDELLLYEEFI (SEQ ID NO:111). 
     
     
         9 . The complex of  claim 1 , wherein the MRD is fused to a heterologous protein. 
     
     
         10 . The complex of  claim 1 , which further comprises an antibody. 
     
     
         11 . The complex of  claim 10 , wherein the MRD and the antibody are covalently bound. 
     
     
         12 . The complex of  claim 11 , wherein the antibody binds ANG-2. 
     
     
         13 . The complex of  claim 11 , wherein the MRD and the antibody bind different targets. 
     
     
         14 . The complex of  claim 13 , wherein the antibody binds a target selected from: VEGF, EGF, IGF-1, FGF1, FGF2, FGF3, FGF4, FGFR1, FGFR2, FGFR3, VEGFR1, EGFR, PDGFR, ErbB2, ErbB3, IGF-1R, cMET, CD19, and CD20. 
     
     
         15 . The complex of  claim 14 , wherein the antibody competitively inhibits:
 binding of trastuzumab to ErbB2;   binding of pertuzumab to ErbB2;   binding of bevacizumab to VEGF;   binding of cetuximab to EGFR;   binding of panitumumab to EGFR;   binding of zalutumumab to EGFR;   binding of nimotuzumab to EGFR;   binding of matuzumab to EGFR;   binding of figitumumab to IGF1R;   binding of AMG 479 to IGF1R;   binding of cixutumumab to IGF1R;   binding of dalotuzumab to IGF1;   binding of BIIB022 to IGF1; or   binding of MEDI-573 to IGF1.   
     
     
         16 . The complex of  claim 13 , wherein the antibody binds a target selected from: interferon-alpha, interferon alpha receptor, interferon beta, interferon beta receptor, interferon-gamma, S1PR, integrin avb3, IL-1B, IL-2, IL-4, IL-4R, IL-5, IL-5R, IL-6, IL-6R, IL-7, IL-8, IL-9, IL-9R, IL-10R, IL-12, IL-13, IL-23, IL-15, IL-18, IL-21, ICOS. PD1, and LIF. 
     
     
         17 . The complex of  claim 13 , wherein the antibody binds TNF. 
     
     
         18 . The complex of  claim 17 , wherein the antibody competitively inhibits binding of adalimumab, golumimab, or infliximab to TNF. 
     
     
         19 . The complex of  claim 11 , which further comprises an additional MRD capable of binding a target other than ANG-2. 
     
     
         20 . The complex of  claim 19 , wherein the MRD binds to a target selected from: VEGF, EGF, IGF-1, FGF1, FGF2, FGF3, FGF4, FGFR1, FGFR2, FGFR3, VEGFR I, EGFR, PDGFR, ErbB2, ErbB3, IGF-1R, cMET, CD19, CD20, TNF alpha, IL-6, interferon-alpha, interferon alpha receptor, interferon beta, interferon beta receptor, interferon-gamma, S1PR, integrin avb3, IL-1B, IL-2, IL-4, IL-4R, IL-5, IL-5R, IL-6, IL-6R, IL-7, IL-8, IL-9. IL-9R, IL-10R, IL-11, IL-12, IL-13, IL-23, IL-15, IL-18, IL-21, ICOS, PD1, and LIF. 
     
     
         21 . A polynucleotide encoding a polypeptide fusion comprising the MRD of  claim 1 . 
     
     
         22 . The polynucleotide of  claim 21  wherein the MRD is fused to:
 the amino terminus of an antibody heavy chain; 
 the amino terminus of an antibody light chain; 
 the carboxyl terminus of an antibody heavy chain; or 
 the carboxyl terminus of an antibody light chain. 
 
     
     
         23 . A vector comprising the polynucleotide of  claim 21 . 
     
     
         24 . A host cell comprising the vector of  claim 23 . 
     
     
         25 . A pharmaceutical composition comprising the complex of  claim 1 . 
     
     
         26 . A method for producing an MRD capable of binding ANG-2, the method comprising culturing the host cell of  claim 24  under conditions wherein the nucleotide sequence encoding the MRD is expressed as a protein and recovering said protein. 
     
     
         27 . A method for treating a patient having an inflammatory disorder comprising administering to said patient a therapeutically effective amount of the complex of  claim 1 . 
     
     
         28 . The method of  claim 27 , wherein the inflammatory disorder is an autoimmune disease or an inflammatory bowel disease. 
     
     
         29 . The method of  claim 28 , wherein the inflammatory bowel disease is Crohn's disease. 
     
     
         30 . A method for treating a patient having arthritis comprising administering to said patient a therapeutically effective amount of the complex of  claim 1 . 
     
     
         31 . The method of  claim 30 , wherein the arthritis is rheumatoid arthritis, juvenile idiopathic arthritis, or psoriatic arthritis.