US2022089730A1PendingUtilityA1

Antigen binding constructs to target molecules

64
Assignee: IMAGINAB INCPriority: Aug 7, 2015Filed: Nov 15, 2021Published: Mar 24, 2022
Est. expiryAug 7, 2035(~9.1 yrs left)· nominal 20-yr term from priority
C07K 2317/53C07K 2317/622A61K 2039/505C07K 16/3069C07K 16/30C07K 16/2815A61K 39/395C07K 16/2809A61P 35/00C07K 16/2803A61K 38/00C07K 14/005A61K 47/6849
64
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Claims

Abstract

Antigen binding constructs that bind to desired targets are disclosed, for example antibodies, including antibody fragments (such as scFv and minibodies) that bind to a target molecule, and have one or more of the disclosed hinges are described herein. Methods of use are described herein.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . An amino acid hinge region comprising a sequence of SEQ ID NO: 1 (X n1 CX n2 X n3 CX n4 X n5 C), wherein X n1  can be any amino acid that does not naturally form a covalent crosslinking bond, wherein X n2  is one of: A, R, N, D, E, Q, G, H, I, L, K, M, F, P, S, T, W, Y, or V, wherein X n3  can be any amino acid, wherein X n4  can be any amino acid, and wherein X n5  can be any amino acid. 
     
     
         2 . The amino acid hinge region of  claim 1 , wherein X n1  does not form a covalent crosslinking bond with another amino acid (SEQ ID NO: 191). 
     
     
         3 . The amino acid hinge region of  claim 1 , wherein X n1  is not a cysteine (SEQ ID NO: 192). 
     
     
         4 . The amino acid hinge region of  claim 3 , wherein X n1  is one of: A, R, N, D, E, Q, G, H, I, L, K, M, F, P, S, T, W, Y, or V (SEQ ID NO: 193). 
     
     
         5 . The amino acid hinge region of  claim 3 , wherein X n2  is P, V, or E (SEQ ID NO: 194). 
     
     
         6 . The amino acid hinge region of  claim 3 , wherein X n2  is P or V (SEQ ID NO: 195). 
     
     
         7 . The amino acid hinge region of  claim 5 , wherein X n4  is P, V, or E (SEQ ID NO: 196). 
     
     
         8 . The amino acid hinge region of  claim 3 , wherein X n4  is P or V (SEQ ID NO: 197). 
     
     
         9 . The amino acid hinge region of  claim 3 , wherein X n3  is P or E (SEQ ID NO: 198). 
     
     
         10 . The amino acid hinge region of  claim 3 , wherein X n5  is P or E (SEQ ID NO: 199). 
     
     
         11 . The amino acid hinge region of  claim 9 , wherein X n3  is P or E (SEQ ID NO: 200). 
     
     
         12 . The amino acid hinge region of  claim 3 , wherein X n2 X n3  is VE (SEQ ID NO: 201). 
     
     
         13 . The amino acid hinge region of  claim 3 , wherein X n2 X n3  is PP (SEQ ID NO: 202). 
     
     
         14 . The amino acid hinge region of  claim 3 , wherein X n4 X n5  is VE (SEQ ID NO: 203). 
     
     
         15 . The amino acid hinge region of  claim 3 , wherein X n4 X n5  is PP (SEQ ID NO: 204). 
     
     
         16 . The amino acid hinge region of  claim 3 , wherein X n2 X n3  is VE and X n4 X n5  is PP (SEQ ID NO: 205). 
     
     
         17 . The amino acid hinge region of  claim 3 , wherein X n2 X n3  is PP and X n4 X n5  is PP or VE (SEQ ID NO: 206). 
     
     
         18 . The amino acid hinge region of  claim 3 , wherein X n2 X n3  is VE and X n4 X n5  is VE or PP (SEQ ID NO: 207). 
     
     
         19 . The amino acid hinge region of  claim 3 , further comprising an extension or lower hinge sequence C-terminal to the last cysteine in SEQ ID NO: 1. 
     
     
         20 . The amino acid hinge region of  claim 19 , wherein the extension or lower hinge sequence comprises at least one of S, G, A, P, or V. 
     
     
         21 . The amino acid hinge region of  claim 20 , wherein the extension sequence comprises at least GGGSSGGGSG (SEQ ID NO: 59). 
     
     
         22 . The amino acid hinge region of  claim 20 , wherein a linker sequence comprises at least APPVAGP (SEQ ID NO: 60). 
     
     
         23 . The amino acid hinge region of  claim 1 , wherein the hinge region of  claim 1  is part of a core hinge region. 
     
     
         24 . The amino acid hinge region of  claim 23 , further comprising an upper hinge region adjacent to the core hinge region. 
     
     
         25 . The amino acid hinge region of  claim 23 , further comprising a lower hinge or extension region adjacent to the core hinge region. 
     
     
         26 . The amino acid hinge region of  claim 25 , further comprising an upper hinge region adjacent to the core hinge region. 
     
     
         27 . The amino acid hinge region of  claim 1 , wherein X n1  comprises a serine, a threonine, or an alanine (SEQ ID NO: 209). 
     
     
         28 . The amino acid hinge region of  claim 1 , wherein X n1  comprises a serine (SEQ ID NO: 210). 
     
     
         29 . The amino acid hinge region of  claim 1 , wherein X n1  comprises an alanine (SEQ ID NO: 211). 
     
     
         30 . The amino acid hinge region of  claim 1 , wherein the amino acid hinge region comprises at least one of the following sequences: SCVECPPCP (SEQ ID NO: 56) or TCPPCPPC (SEQ ID NO: 166). 
     
     
         31 . The amino acid hinge region of  claim 1 , wherein the amino acid hinge region comprises at least one of the following sequences: ERKSCVECPPCP (SEQ ID NO: 167), EPKSSDKTHT (SEQ ID NO: 46), and CPPCPPC (SEQ ID NO: 52). 
     
     
         32 . The amino acid hinge region of  claim 1 , wherein the amino acid hinge region comprises at least one of the following sequences: ERKSCVECPPCPGGGSSGGGSG (SEQ ID NO: 34) or ERKSCVECPPCPAPPVAGP (SEQ ID NO: 33) or EPKSSDKTHTCPPCPPCGGGSSGGGSG (SEQ ID NO: 26) or EPKSSDKTHTCPPCPPCAPELLGGP (SEQ ID NO: 25). 
     
     
         33 . An amino acid hinge region comprising a sequence of SEQ ID NO: 2 (X n1  X n2  X n3  X n4 X n5  X n6 CX n7 X n8 CX n9 X n10 C), wherein X n1  can be any m amino acids, wherein m is any number of amino acids of any type, wherein X n2  can be any amino acid, wherein X n3  can be any amino acid, wherein X n4  can be any amino acid, wherein X n5  can be any amino acid, wherein X n6  can be any amino acid other than a cysteine, wherein X 17  can be any amino acid, wherein X n8  can be any amino acid, wherein X n9  can be any amino acid, and wherein X n10  can be any amino acid. 
     
     
         34 . The amino acid hinge region of  claim 33 , wherein X n1  is not a cysteine (SEQ ID NO: 213). 
     
     
         35 . The amino acid hinge region of  claim 33 , wherein X n2  is not a cysteine (SEQ ID NO: 214). 
     
     
         36 . The amino acid hinge region of  claim 33 , wherein X n2  is a D (SEQ ID NO: 215). 
     
     
         37 . The amino acid hinge region of  claim 33 , wherein X n3  is a K (SEQ ID NO: 216). 
     
     
         38 . The amino acid hinge region of  claim 33 , wherein X n4  is a T (SEQ ID NO: 217). 
     
     
         39 . The amino acid hinge region of  claim 33 , wherein X n5  is an H (SEQ ID NO: 218). 
     
     
         40 . The amino acid hinge region of  claim 33 , wherein X n6  is a T (SEQ ID NO: 219). 
     
     
         41 . The amino acid hinge region of  claim 33 , wherein X n1  is a P or a V (SEQ ID NO: 220). 
     
     
         42 . The amino acid hinge region of  claim 33 , wherein X n8  is a P or an E (SEQ ID NO: 221). 
     
     
         43 . The amino acid hinge region of  claim 33 , wherein X n9  is a P or a V (SEQ ID NO: 222). 
     
     
         44 . The amino acid hinge region of  claim 33 , wherein X n10  is a P or an E (SEQ ID NO: 223). 
     
     
         45 . The amino acid hinge region of  claim 33 , further comprising a CXXC (SEQ ID NO: 224) or CXXC (SEQ ID NO: 225) motif that is positioned in front of X 1 . 
     
     
         46 . The amino acid hinge region of  claim 33 , further comprising a X n11 X n12 C sequence immediately attached to the C-terminal cysteine in SEQ ID NO: 1, wherein X 11  can be any amino acid, and wherein X n12  can be any amino acid (SEQ ID NO: 244). 
     
     
         47 . The amino acid hinge region of  claim 46 , wherein X n1  is a P or a V, and wherein X n12  is a P or an E (SEQ ID NO: 227). 
     
     
         48 . The amino acid hinge region of  claim 33 , wherein X n1  is a serine, X n2  is a D, X n3  is a K, X n4  is a T, X n5  is an H, X n6  is a T, X n7  is a P, X n8  is a P, X 19  is a P, and X n10  is a P (SEQ ID NO: 228). 
     
     
         49 . The amino acid hinge region of  claim 33 , wherein the hinge region comprises at least one of the following sequences: CPPCPPC (SEQ ID NO: 52), CPPCVECPPC (SEQ ID NO: 53), or CPPCPPCPPC (SEQ ID NO: 54). 
     
     
         50 . The amino acid hinge region of  claim 33 , wherein the hinge region comprises at least one of the following sequences: EPKSSDKTHTCPPCPPC (SEQ ID NO: 168), EPKSSDKTHTCPPCVECPPC (SEQ ID NO: 169), or EPKSSDKTHTCPPCPPCPPC (SEQ ID NO: 170). 
     
     
         51 . The amino acid hinge region of  claim 33 , wherein the hinge region comprises at least one of the following sequences: EPKSSDKTHTCPPCPPCGGGSSGGGSG (SEQ ID NO: 26), EPKSSDKTHTCPPCVECPPCGGGSSGGGSG (SEQ ID NO: 28), or EPKSSDKTHTCPPCPPCPPCGGGSSGGGSG (SEQ ID NO: 30). 
     
     
         52 . An amino acid hinge region comprising:
 a core hinge sequence of at least one of: CVECPPCP (SEQ ID NO: 57), CPPCPPC (SEQ ID NO: 52), or CPPCPPCPPC (SEQ ID NO: 54), or CPPCVECPPC (SEQ ID NO: 53) linked to;   an upper hinge sequence of ELKTPLGDTTHT (SEQ ID NO: 48) or EPKSSDKTHT (SEQ ID NO: 46).   
     
     
         53 . An amino acid hinge region for an antibody comprising:
 an upper hinge region that comprises no amino acids capable of crosslinking with a corresponding amino acid; and   a core hinge region connected to a C-terminus of the upper hinge region, wherein the core hinge region comprises at least three cysteines per strand.   
     
     
         54 . The amino acid hinge region of 52, wherein the amino acid hinge region further comprises a lower hinge or extension region connected C-terminal to the core hinge region, wherein the lower hinge or extension sequence is at least one of: APPVAGP (SEQ ID NO: 60), APELLGGP (SEQ ID NO: 58), and/or GGGSSGGGSG (SEQ ID NO: 59). 
     
     
         55 . The amino acid hinge region of 52, wherein the upper hinge region comprises no cysteines that crosslink within the upper hinge region. 
     
     
         56 . The amino acid hinge region of  claim 52 , wherein the upper hinge region comprises no cysteines. 
     
     
         57 . The amino acid hinge region of  claim 52 , further comprising a lower hinge or extension region. 
     
     
         58 . The amino acid hinge region of  claim 52 , wherein the lower hinge or extension region comprises at least one of: GGGSSGGGSG (SEQ ID NO: 59) or APPVAGP (SEQ ID NO: 60) or APELLGGP (SEQ ID NO: 58). 
     
     
         59 . The amino acid hinge region of  claim 1 , wherein when located within a minibody, and wherein when the minibody is administered to a human subject, clearance of the minibody from the subject occurs primarily through a liver. 
     
     
         60 . The amino acid hinge region of  claim 1 , wherein, when located within a minibody, and wherein when the minibody is administered to a human subject, clearance of the minibody from the subject does not occur primarily through a kidney. 
     
     
         61 . The amino acid hinge region of  claim 1 , wherein the hinge region is within an antibody. 
     
     
         62 . The amino acid hinge region of  claim 1 , wherein the hinge region is within an antibody binding fragment. 
     
     
         63 . The amino acid hinge region of  claim 1 , wherein the hinge region is within a minibody. 
     
     
         64 . The amino acid hinge region of  claim 1 , wherein the hinge region is within a monospecific antibody. 
     
     
         65 . The amino acid hinge region of  claim 1 , wherein the hinge region comprises at least three cysteines per strand. 
     
     
         66 . The amino acid hinge region of  claim 1 , wherein the hinge region comprises at least four cysteines per strand. 
     
     
         67 . The amino acid hinge region of  claim 1 , wherein the hinge region comprises at least five cysteines per strand. 
     
     
         68 . The amino acid hinge region  claim 65 , wherein cysteines are distributed throughout the amino acid hinge region in a repeating CXX or CXY motif. 
     
     
         69 . The amino acid hinge region of  claim 1 , wherein the hinge region is within a bispecific antibody. 
     
     
         70 . The amino acid hinge region of  claim 69 , wherein the bispecific antibody is assembled in a 1:1 ratio. 
     
     
         71 . The amino acid hinge region of  claim 69 , wherein the bispecific antibody comprises an antibody fragment. 
     
     
         72 . The amino acid hinge region of  claim 71 , wherein the bispecific antibody is a minibody. 
     
     
         73 . A pharmaceutical composition comprising the amino acid hinge region of  claim 1 , wherein less than 5% aggregation of an antibody is present in the composition. 
     
     
         74 . A pharmaceutical composition comprising the amino acid hinge region of  claim 1 . 
     
     
         75 . The pharmaceutical composition of  claim 73 , wherein at least 1 microgram to 100 mg of the antibody is present. 
     
     
         76 . A minibody comprising a core hinge region, wherein the core hinge region comprises at least three cysteines per strand forming at least three disulfide bonds within the core hinge region. 
     
     
         77 . The minibody of  claim 76 , wherein the first residue of the core region is a serine. 
     
     
         78 . The minibody of  claim 76 , wherein the core hinge region comprises SCVECPPCP (SEQ ID NO: 56). 
     
     
         79 . A minibody comprising a sequence X n1 CX n2 X n3 CX n4 X n5 C (SEQ ID NO: 3), wherein SEQ ID NO: 3 is located as the core hinge region of the minibody, and wherein X n1  can be any amino acid or no amino acid, X n2  can be any amino acid, X n3  can be any amino acid, X n4  can be any amino acid, and X n5  can be any amino acid. 
     
     
         80 . The minibody of  claim 79 , wherein X n1  is any amino acid other than a cysteine (SEQ ID NO: 229). 
     
     
         81 . The minibody of  claim 79 , wherein X n1  is a serine (SEQ ID NO: 230). 
     
     
         82 . A variant minibody hinge comprising:
 a first altered amino acid position, wherein the first altered position is an amino acid that in a native antibody hinge would be a cysteine, and has been altered in the first altered position so that it does not form a disulfide bond; and   at least three cysteines per strand C-terminal to the first altered amino acid position.   
     
     
         83 . The amino acid hinge region of  claim 1 , wherein the hinge region consists of SEQ ID NO: 1. 
     
     
         84 . The amino acid hinge region of  claim 1 , wherein SEQ ID NO: 1 is a core hinge region, and wherein the core hinge region essentially consists of SEQ ID NO: 1. 
     
     
         85 . The amino acid hinge region of  claim 84 , wherein the core hinge region consists of SEQ ID NO: 1.

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