Anticolon Cancer Activity Of Largazole Pharmacokinetics In The Rats By LC/MS/MS, And Clinical Pharmacokinetics Of L-Tetrahydropalmatine With Cocaine Exposure In People With A History Of Cocaine Use

1. ANTICOLON CANCER ACTIVITY OF LARGAZOLE PHARMACOKINET-ICS IN THE RATS BY LC/MS/MSLargazole as histone deacetylases (HDACs) inhibitor, a 16-membered cyclic peptide lactones compound, was isolated from a marine cyanobacterium by Dr. Luesch lab in University of Flor-ida. Largazole as a prcodrug can rapidly be hydrolysed and release a potent HDACs inhibitor, Largazole Thiol. Largazole can chelate Zn2 in the active site HDACs to inhibit HDACs. Previous studies showed that Largazole has good anti-colon cancer activity on cells and mouse models. This project was designed to evaluate the pharmacokinetic of Largazole in rats and provide sci-entidic data for the new drug candidate development and clinical study. The details of this pro-ject are as follows:1. The determination and quanitative analytical method of Largazole Thiol in plasma by LC/MS/MS.Developed a new plasma proceeding and analytical method to analysis Largazole Thiol in plas-ma using LC/MS/MS. The method was validated according to guidance for industry bioanalyti-cal method validation of U.S. Food and Drug Administration. The results showed that Largazole Thiol in plasma concentration range of 12.5ng/mL to 400ng/mL has good linear relationship, and the regression equation was y= 0.003x+0.0076, R2= 0.9997.The LLOQ of the method was 12.5ng/mL; the intra-day precision for quality controls were range from 2.56% to 12.08%; the inter-day precision were range from 7.12% to 9.77%; the recovery of the method were range from 77% to 87%; the matrix effects were range from 72% to 75%; the plasma samples were stable under the condition of the short-term, frozen-thaw for once, twice, and three times; all the characters of the analytical method were meet the requirement of Food and Drug Administration.2. The plasma protein binding of Largazole Thiol The plasma protein binding of Largazole Thiol in human and rat plasma as determined by ultra-filtration method were 90.13% and 77.14%, respectively.3. Pharmacokinetic study of Largazole in rats According to the concentration time curve of Largazole Thiol in rats’plasma after the i.v. admin-istration at 10 mg/kg, the half-time of Largazole in rats was 0.5 hr analyzed by noncompartmen-tal model.Compared one-compartmemtal model and two-compartmental model for Largazole using phoe-nix and NONMEM software, the results showed that the two-compartmental model was better fit than one compartmental model. And the model was validated by NONMEM.2. CLINICAL PHARMACOKINETICS OF L-TETRAHYDROPALMATINE WITH COCAINE EXPOSURE IN PEOPLE WITH A HISTORY OF COCAINE USEL-tetrahydropalmatine as the active substance was isolated from traditional chinse medicine, co-rydalis. L-THP has been as analgesics (rotunine) in clinical practice in china for 40 years. Recent studies showed that L-THP has good anti-drug addiction. This unique pharmacological profile makes 1-THP an excellent candidate for medication development for the treatment of cocaine addiction. This study is to evaluate the clinical pharmacokinetic and safty of L-THP, and provide the scientific data for future clinical study and its dosage regiman. The details of this project are as follows:1. The determination and quantitative analytical method of L-THP and cocaineDeveloped the new method to analysis and quantitate L-THP and cocaine in plasma using UPLC-FLD, and the method was validated according to the guidance for industry bioanalytical method validation of U.S. Food and Drug Administration. The results showed that the characters of the method, such as selectitvity, standard curve, lower limit of quantification, intra-day presi- cion and accuracy, inter-day presicion and accuracy, recovery, and stability, met the require-ments of U.S. Food and Drug Administration.2. The clinical pharmacokinetic study of L-THP and cocaineThe pharmacokinetic parameters of L-THP and cocaine were deterimined by plasma concentra-tion-time cure on patients after drug administration using non-compartmental model.3. The clinical safety studies of L-THPComparing the blood pressure and heart rate after cocaine administration between L-THP group and placebo group, the results showed that the blood pressure and heart rate patients in L-THP group were no significantly increased, and meet the safety requirement of the U.S. Food and Drug Administration.