US 12,298,319 B2
Method for measuring alpha value of muscarinic M1 receptor positive allosteric modulators
Haruhide Kimura, Kanagawa (JP); Yuu Sako, Kanagawa (JP); Satoru Matsuda, Kanagawa (JP); Yuji Shimizu, Kanagawa (JP); Emi Kurimoto, Kanagawa (JP); and Takao Mandai, Kanagawa (JP)
Assigned to Takeda Pharmaceutical Company Limited, Osaka (JP)
Appl. No. 16/614,588
Filed by Takeda Pharmaceutical Company Limited, Osaka (JP)
PCT Filed May 17, 2018, PCT No. PCT/JP2018/019208
§ 371(c)(1), (2) Date Nov. 18, 2019,
PCT Pub. No. WO2018/212312, PCT Pub. Date Nov. 22, 2018.
Claims priority of provisional application 62/508,609, filed on May 19, 2017.
Prior Publication US 2020/0182850 A1, Jun. 11, 2020
Int. Cl. G01N 33/94 (2006.01); A61K 45/06 (2006.01); G01N 33/15 (2006.01)
CPC G01N 33/944 (2013.01) [G01N 33/15 (2013.01); A61K 45/06 (2013.01); G01N 2333/4703 (2013.01); G01N 2500/04 (2013.01); G01N 2500/20 (2013.01)] 12 Claims
 
1. A method for screening for a cholinergic muscarinic M1 receptor (M1R) positive allosteric modulator (M1PAM) with reduced cholinergic side effects, comprising the following steps (a) to (c):
(a) contacting M1R or a partial peptide thereof with a test compound, acetylcholine (ACh) and detectable M1 antagonist,
(b) measuring α value of the test compound, wherein the α value is calculated as αAB from numeric formula (ii):

OG Complex Work Unit Math
wherein Y is the percentage of specific binding;
[A], [B], and [C] are respectively the concentration of ACh, the test compound and detectable M1 antagonist;
KA, KB and Kc respectively show the equilibrium dissociation constant of ACh, the test compound and detectable M1 antagonist; and
αAB and αBC are respectively the cooperativities between the test compound and ACh, and the test compound and detectable M1 antagonist,
(c) repeating steps (a) and (b) to screen one or more test compounds and selecting one or more of the test compounds with a low α value, each as an MIPAM, wherein the low a value is associated with reduced cholinergic side effects, and wherein the low α value is greater than 1 and not more than 500, and wherein the M1PAM with reduced cholinergic side effects has not less than 2-fold safety margin between a minimal cognition effective dose and a minimal cholinergic side effect dose in mammals.