Method for determining the stability of organic methyleneamines in the presence of semicarbazide-sensitive amine oxidase

摘要

The present invention provides methods for determining the stability of methyleneamine, methyleneamine-like compounds or compounds containing a methyleneamine moiety in the presence of semicarbazide-sensitive amine oxidase (SSAO) or a biological sample containing SSAO activity. The disclosed methods may be configured in an assay format for high throughput screening applications.

主权项

1. A method of determining the relative metabolic stability profile of a test agent due to semicarbazide-sensitive amine oxidase (SSAO) catalyzed metabolism in recombinant cultured cells expressing full-length SSAO and in a biological sample comprising SSAO, wherein the test agent is a drug candidate and the metabolic stability of the test agent due to SSAO catalyzed metabolism is unknown, the method comprising:
a) determining the metabolic stability of the test agent in the presence of recombinant cultured cells expressing full-length SSAO; by:
i) adding the test agent to the recombinant cultured cells;ii) incubating the test agent with the recombinant cultured cells for a predetermined period of time;iii) measuring the amount of the test agent remaining in the presence of the recombinant cultured cells at the predetermined period of time; andiv) comparing the amount of the test agent remaining in the presence of the recombinant cultured cells at the predetermined period of time to a control to determine the metabolic stability of the test agent in the presence of recombinant cultured cells expressing full-length SSAO; b) determining the metabolic stability of the test agent in the presence of a biological sample comprising SSAO by:
i) adding the test agent to the biological sample;ii) incubating the test agent with the biological sample for a predetermined period of time;iii) measuring the amount of the test agent remaining in the presence of the biological sample at the predetermined period of time; andiv) comparing the amount of the test agent remaining in the presence of the biological sample at the predetermined period of time to a control to determine the metabolic stability of the test agent in the presence of a biological sample comprising SSAO; and c) comparing the metabolic stability of the test agent in the presence of recombinant cultured cells expressing full-length SSAO to the metabolic stability of the test agent in the presence of a biological sample comprising SSAO to determine the relative metabolic stability profile of the test agent.