US 11,713,358 B2
Chimeric polypeptide assembly and methods of making and using the same
Volker Schellenberger, Palo Alto, CA (US); Fan Yang, San Jose, CA (US); Desiree Thayer, Burlingame, CA (US); Bee-Cheng Sim, Mountain View, CA (US); and Chia-Wei Wang, Santa Clara, CA (US)
Assigned to AMUNIX PHARMACEUTICALS, INC., South San Francisco, CA (US)
Appl. No. 15/753,716
Filed by AMUNIX PHARMACEUTICALS, INC., South San Francisco, CA (US)
PCT Filed Aug. 26, 2016, PCT No. PCT/US2016/049137
§ 371(c)(1), (2) Date Feb. 20, 2018,
PCT Pub. No. WO2017/040344, PCT Pub. Date Mar. 9, 2017.
Claims priority of provisional application 62/379,673, filed on Aug. 25, 2016.
Claims priority of provisional application 62/363,046, filed on Jul. 15, 2016.
Claims priority of provisional application 62/338,285, filed on May 18, 2016.
Claims priority of provisional application 62/278,755, filed on Jan. 14, 2016.
Claims priority of provisional application 62/263,319, filed on Dec. 4, 2015.
Claims priority of provisional application 62/211,532, filed on Aug. 28, 2015.
Prior Publication US 2019/0153115 A1, May 23, 2019
Int. Cl. C07K 16/30 (2006.01); C07K 16/28 (2006.01); A61P 35/00 (2006.01); C07K 14/00 (2006.01); A61K 39/00 (2006.01); A61K 38/00 (2006.01)
CPC C07K 16/30 (2013.01) [A61P 35/00 (2018.01); C07K 14/00 (2013.01); C07K 16/2809 (2013.01); A61K 38/00 (2013.01); A61K 2039/505 (2013.01); A61K 2039/54 (2013.01); A61K 2039/545 (2013.01); A61K 2039/572 (2013.01); C07K 2317/24 (2013.01); C07K 2317/31 (2013.01); C07K 2317/56 (2013.01); C07K 2317/565 (2013.01); C07K 2317/60 (2013.01); C07K 2317/622 (2013.01); C07K 2317/73 (2013.01); C07K 2317/732 (2013.01); C07K 2317/76 (2013.01); C07K 2317/90 (2013.01); C07K 2317/92 (2013.01); C07K 2317/94 (2013.01); C07K 2319/01 (2013.01); C07K 2319/035 (2013.01); C07K 2319/21 (2013.01); C07K 2319/30 (2013.01); C07K 2319/31 (2013.01); C07K 2319/50 (2013.01)] 72 Claims
 
1. A chimeric polypeptide assembly comprising a first portion, a second portion, and a third portion wherein:
a. the first portion comprises:
i. a first binding domain with binding specificity to a target cell marker; and
ii. a second binding domain with binding specificity to an effector cell antigen;
b. the second portion comprises a peptidyl release segment (RS) capable of being cleaved by one or more mammalian proteases;
c. the third portion comprises an extended recombinant polypeptide, wherein the extended recombinant polypeptide is capable of being released from the first portion by action of the mammalian protease on the second portion; and
d. the assembly is a monomeric fusion protein configured, in an N- to C-terminus orientation, of binding domains-RS- extended recombinant polypeptide or extended recombinant polypeptide -RS-binding domains,
wherein the chimeric polypeptide assembly is characterized by the ratio between 1) relative cytotoxicity and 2) relative effector-cell antigen binding affinity multiplied with relative binding affinity to the tumor specific marker, being greater than at least 3:1, wherein:
a) relative cytotoxicity is measured as a ratio between cytotoxicity of (i) the released first portion to the target cell in an in vitro assay comprising both the effector cells bearing the effector-cell antigen and tumor cells bearing the target cell marker and (ii) a composition comprising a corresponding first portion of the chimeric polypeptide assembly and a corresponding third portion of the chimeric polypeptide assembly linked by a non-cleavable peptide of 1 to about 10 amino acids;
b) relative effector cell antigen binding affinity, is measured as a ratio between (i) binding affinity of the released first portion to the effector cell antigen and (ii) the binding affinity of a composition comprising the corresponding first portion of the chimeric polypeptide assembly and the corresponding third portion of the chimeric polypeptide assembly linked by a non-cleavable peptide of 1 to about 10 amino acids to the effector cell antigen; and
c) relative target-cell marker binding affinity is measured as a ratio between (i) the binding affinity of the released first portion to the target-cell marker and (ii) the binding affinity to the target-cell marker of a composition comprising the corresponding first portion of the chimeric polypeptide assembly and the corresponding third portion of the chimeric polypeptide assembly linked by a non-cleavable peptide of 1 to about 10 amino acids.