Chemical Components Analysis Of Radix Cudramiae And Its Inhibitory Effects On CYP450 Enzymes

Radix cudramiae is a traditional Chinese herbal medicine.It exhibited a broad spectrum of pharmacological activities,including anti-inflammatory,anti-oxidation,hepatoprotective and anti-tumor activities.In this work,the chemical components of Radix cudramiae were investigated by UPLC-QTOF-MS and GC-MS.The in vitro inhibitory activities of Radix cudramiae extract on human liver microsomal CYP450enzymes were also investigated.Firstly,the preparation of Radix cudramiae was investigated.The optimized extraction conditions were as below:the extraction ratio was 1?50 g/m L using 60%methanol as the extraction solvent,and the ultrasonic extraction was performed at35℃for 45 min.The optimized parameters of the ball milling were 100 rpm for 10min with ball milling filling rate of 5.4%.The UPLC method was established and optimized to separate all the components in Radix cudramiae extracts within 50 min.For the separation and purification of oxyresveratrol from Radix cudramiae extracts the suitable macroporous adsorption resin were screened.Secondly,UPLC-QTOF-MS method was established to identify the chemical components in Radix cudramiae extracts,and 36 compounds including xanthones,organic acids,flavones,stilbenes and other compounds were tentatively assigned.GC-MS combined with retention index were used to analyze the volatile components in Radix cudramiae,and 65 volatile components were identified,including 33 fatty components,21 terpenoid components and 11 aromatic compounds,accounting for50.8%,32.3%and 16.9%of the total components,respectively.Thirdly,the inhibitory activities of Radix cudramiae on human liver microsomal CYP450 enzymes were investigated.The Radix cudramiae extracts was incubated with mixed probe drugs,and the peak areas of each probe drug were determined by UPLC with compared to the positive inhibitors.Results showed that the IC₅₀values of Radix cudramiae extracts for CYP1A2,CYP3A4,CYP2C9 and CYP2E1 were 0.3561,0.3324,0.4049 and 0.8615 mg/m L,respectively.And the IC₅₀values of oxyresveratrol for CYP1A2,CYP3A4,CYP2C9 and CYP2E1 were 106.4,135.1,125.8 and 125.6μM,respectively.For the four subtypes of human liver microsomal CYP450 enzymes,Radix cudramiae extraction showed the largest inhibition effects on CYP3A4 subtype enzyme,and the smallest inhibition effects on CYP2E1.And oxyresveratrol showed the largest inhibition effects on CYP1A2 subtype enzyme,and the smallest inhibition effects on CYP3A4.Compared with the positive inhibitors,the Radix cudramiae exhibited weak inhibitory activity on these four subtype enzymes.