US 9,810,690 B2
Method for screening inhibitors of Ras
Matthew P. Patricelli, San Diego, CA (US); Ulf Peters, San Diego, CA (US); Liansheng Li, San Diego, CA (US); Pingda Ren, San Diego, CA (US); and Yi Liu, San Diego, CA (US)
Assigned to ARAXES PHARMA LLC, San Diego, CA (US)
Filed by Araxes Pharma LLC, La Jolla, CA (US)
Filed on Nov. 2, 2016, as Appl. No. 15/342,100.
Application 15/342,100 is a continuation in part of application No. PCT/US2016/057774, filed on Oct. 19, 2016.
Claims priority of provisional application 62/243,439, filed on Oct. 19, 2015.
Prior Publication US 2017/0131278 A1, May 11, 2017
Int. Cl. C12N 9/14 (2006.01); G01N 33/573 (2006.01); C12N 9/00 (2006.01); C12N 15/70 (2006.01)
CPC G01N 33/573 (2013.01) [C12N 9/14 (2013.01); C12Y 306/05002 (2013.01); G01N 2500/04 (2013.01); G01N 2500/20 (2013.01)] 30 Claims
 
1. A method of selecting a Ras antagonist, the method comprising:
(a) combining in a reaction mixture a mutant Ras, a competition probe, and a test compound; and
(b) detecting a decrease in binding between the mutant Ras and the competition probe as compared to binding of the competition probe to the mutant Ras in an absence of the test compound; wherein:
i. the mutant Ras comprises a truncated or full-length sequence according to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4, that is mutated to have up to 20 mutations including mutation of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4 at amino acid residue 62, 92, or 95 to cysteine;
ii. the competition probe is capable of binding and covalently modifying the mutant Ras; and
iii. the decrease in binding between the mutant Ras and the competition probe is indicative of Ras antagonist activity of the test compound.