Pharmaceutical Research Of8-hydroxypiperidine-methyl-baicalein Mesylate As CDK1Inhibitor

Cyclin-dependent kinases (CDKs) play a significant role in cell cycle regulation and have become the new targets in anticancer drugs. A novel derivative of natural baicalein was synthesized by Mannich reaction as CDK1inhibitor, which can inhibit the cell proliferation and selectively induce tumor cells apoptosis and exhibit excellent anticancer activity.8-hydroxypiperidine-methyl-baicalein and its related substances:6-hydroxypiperidine-methyl-wogonin,8-hydroxypiperidine-methyl-oroxylin,8-hydroxypiperidine-methyl-scutellarein,6-hydroxypiperidine-methyl-norwogonin and8-hydroxypiperidine-methyl-chrysin, derivatives of natural skullcapflavones, were synthesized by Mannich reaction and characterized and of which8-hydroxypiperidine-methyl-scutellarein,6-hydroxypiperidine-methyl-norwogonin and8-hydroxypiperidine-methyl-chrysin were new compounds. To elevate the purity of8-hydroxypiperidine-methyl-baicalein,8-hydroxypiperidine-methyl-baicalein mesylate was synthesized. The preparing process was optimized and stable, the average yield was91.8%and the average content was99.1%.A simple and reliable reversed phase liquid chromatographic method has been developed and subsequently validated for assay of8-hydroxypiperidine-methyl-baicalein mesylate. Assay of8-hydroxypiperidine-methyl-baicalein mesylate was achieved with LP-C18column, methanol-isopropyl alcohol-water-methane sulfonic acid (43:2:55:0.2, v/v/v/v) as eluent, and UV detection at277nm. The resolution was well with R>1.5. The described method for assay of8-hydroxypiperidine-methyl-baicalein mesylate is linear over a range of6.25-200μg/mL, y=176268x+459482(R2=0.9999, n=6). The precision was well (RSD%1.09%, n=6) and the repeatability was well (RSD%1.35%, n=6) with RSD%below2%and consistent with the requirement.A simple and reliable reversed phase liquid chromatographic method has been developed and subsequently validated for determination of related substances. The linearity of8-hydroxypiperidine-methyl-scutellarein,8-hydroxypiperidine-methyl-oroxylin and baicalein was well in the range of1.25-20μg/mL (R2=0.9999,0.9997,0.9998, n=5). The standard recovery were101.9%,100.9%,100.2%with RSD%of3.48%,3.29%,2.34%, respectively, and consistent with the requirement (RSD%<5%).The RSD%of precision were3.42%,2.09%,1.29%, respectively, and consistent with the requirement (RSD%<5%). The limit of quantification was all below0.50%. The average level of the three related substances was all below0.50%, with the other related substances below0.10%.The purity of8-hydroxypiperidine-methyl-baicalein mesylate prepared by the optimized method is above98%, and the content of related substances are below2.0%, consistent with the quality control of new drug.The method of quality evaluation with well specificity, accuracy, precision, repeatability, stability, and robustness is suitable for quality control of8-hydroxypiperidine-methyl-baicalein mesylate.