	US 12,487,235 B2
	Compositions and methods for studying drug metabolism
Tom Sze-Yin Chan, Danbury, CT (US); Alexander Matthew Horspool, Morgantown, WV (US); and Ting Wang, Danbury, CT (US)
Assigned to Boehringer Ingelheim International GmbH, Ingelheim am Rhein (DE)
Filed by Boehringer Ingelheim International GmbH, Ingelheim am Rhein (DE)
Filed on Jan. 14, 2021, as Appl. No. 17/148,748.
Claims priority of provisional application 62/962,333, filed on Jan. 17, 2020.
Prior Publication US 2021/0223237 A1, Jul. 22, 2021
Int. Cl. G01N 33/543 (2006.01); G01N 33/50 (2006.01); G01N 33/58 (2006.01)

CPC G01N 33/54326 (2013.01) [G01N 33/5008 (2013.01); G01N 33/585 (2013.01)]

3 Claims

1. A method for studying drug metabolism, said method comprising:

estimating a proportion of non-specific binding of a small molecule drug by determining a free fraction of the drug unbound to human liver microsomes (HLM), comprising the steps of:

providing a plurality of magnetizable beads in aqueous buffered solution;

providing a plurality of human liver microsomes;

providing the drug;

mixing the plurality of magnetizable beads and plurality of microsomes in the presence of the drug;

applying a magnetic field to the plurality of microsome-beads to separate the plurality of microsome-beads from the aqueous buffered solution;

removing a supernatant from the separated microsome beads;

determining a concentration of drug in the supernatant; and

using the concentration of the drug in the supernatant to calculate the free fraction of the drug unbound to microsomes which is a ratio of the amount of drug in the supernatant divided by an amount of drug in a control sample without added HLM.