Phase Ⅰ Clinical Trial Of RS Capsule And The Gene Polymorphisms Research Of Drug Metabolism Enzymes

Objective:1. To evaluate the tolerance and safety and pharmacokineticscharacteristics of RS capsule to healthy subjects, and provide a basis for theprotocol of phaseâ…¡clinical trial. 2. To investigate the association betweencytochrome P450 enzymes 1A2, UDP-Glucuronosyltransferase IA7 (CYP1A2, UGT1A7)gene polymorphisms and the adverse reaction of the RS capsule. To primarilyestablish method for detecting drug metabolism enzyme gene polymorphisms,analyzing the association between the drug metabolism enzyme genepolymorphisms and new drug clinical trials adverse reaction.Methods:1. The master single dosage tolerance trial:twenty six healthysubjects participated single administration trials for five dosages(10mg,50mg, 100mg, 200mg, 300mg)based on open, randomized, no-control trial method. Atexperimental stage, they take orally RS capsule one time. According to the claimof trial schedule, we observed the safety index and laboratory examinationresults before and after subjects took RS capsule orally. 2. The multipledosage tolerance trial: twelve subjects received multiple administrationtrials for two dosages(200mg and 300mg). One time everyday orally, seven dayscontinuously. We observed the safety index and laboratory examination resultsbefore and after subjects took RS capsule orally. 3. Pharmacokinetics trialsof single administration:thirty subjects recevied single administrationpharmacokinetic trials for three dosages (50mg, 100mg, 300mg). By using open,randomized, parallel trial method, and we studied effect of food on drug by themethod of taking drug orally before and after breakfast(50mg) and blood sampleswere handled by using liquid liquid extraction, we measured plasma drugconcentration by using HPLC/MS/MS. Statistics were analyzed through DAS2.0 pharmacokinetics program. Parameters(C_max/D,AUC/D)were analyzed throughanalysis of variance, T_1/2 were analyzed through rank test. 4. Polymerase chainreaction and restriction fragment length polymorphism techniques were usedto examine the CYP1A2 and UGT1A7 gene polymorphisms of subjects and controls.We analyzed comparatively with the statistical method.Results: 1. rhe master single dosage tolerance trial: Two subjects of10mg-dosage group had thyroid gland function abnormality:one case T4 raisedto 142 nmol·L^-1 on the third day after administration and became normal onthe twelfth day; one subject TSH raised to 5.98μIU·ml^-1 on the third day afteradministration and became normal on the thirtieth day; one case presentedloosestool. In 50mg-dosage group, one subject had distention of head and hadn¹ t chestdistress, dyspnea and anomalopia, blood pressure 86/51mmHg. Urine test: protein(+), after 72h administration, and became normal after three days. In100mg-dosage group, subjects had doubtful adverse events presenting FT4 washigher than the normal level; T wave changed;TSH was higher than the normallevel; erythrocyte appeared in urine test. All these became normal whenrechecked. In 200mg-dosage group, one subject thrombocyte was lower(84×10⁹·L^-1)after 24 hours administration and became normal after 72 hours whenrechecked. In 300mg-dosage group, one subject urine test showed that urineglucose was (++)after 24 hours administration and became normal after twelvedays. 2. The multiple dosage tolerance trial: In 200mg-dosage group, twosubjects had urine test abnormality:one subject had occult blood(+) and fewepithelial cell on the eighth day after administration and became normal onthe tenth day after administration; one subject had few epithelial cell in theurine test on the eighth and ninth day after administration and became normalon the tenth day after administration. In 300mg-dosage group, four subjectshad doubtful and possible adverse events:decreasing slightly of hemoglobin;ALT raised; thyroid gland ultrasonic wave: small nodule in right lobe;ALT andAST arised. 3. Study on pharmacokinetics of RS capsule in healthy people: bya non-compartment model to accumulate the plasma drug concentration data. Cmaxwere (218.69±68.56), (401.99±168:30), (518.53±122.36)μg·L^-1; AUCwere (3325.87±1636.91), (8626.88±4825.04), (10473.61±4326.65)μg·L^-1·h; T_1/2 were(13.01±4.01), (11.77±4.40), (9.71±1.46)h, respectively. Themain pharmacokinetic parameters were similar and had no significant differencefollowing oral administration of 50 and 100mg, Cmax and AUC increased proportionally with increasing dose. This drug showed linear kinetics over thedosage range of 50-100mg. At dose of 300mg, there was a significant statisticaldifference in the main parameters(compared with 50mg).The disposition of thisdrug in vivo did not exhibit linear kinetic characterstics when the. dosagewas 300mg. At dose of 50mg, there was not a significant statistical difference(P＜0.05)in the main parameters between empty stomach and having breakfast, ltsuggested the food was not the factor which effects the drug. 4. The genepolymorphisms research of its relative drug metabolism enzymes: There was nosignificant difference between subjects and controls in genotypes and allelesby X² statistical method. According the response to the trial, divided thesubjects into adverse reaction and non-adverse reaction two groups.CYP1A2G2964A: adverse reaction group had 14 subjects, 10 subjects genotypeswere G/A and covered 71.43%; non-adverse reaction group had 41 subjects, 22subjects genotypes were G/G and covered 53.66%. CYP1A2C734A: adverse reactiongroup had 13 subjects, 7 subjects genotypes were C/A and covered 53.85%;non-adverse reaction group had 32 subjects, 16 subjects genotypes were A/Aand covered 50.00%. UGT1ATTrp208Arg: adverse reaction group had 15 subjects, 12subjects genotypes were Trp/Trp and covered 80.00%; non-adverse reaction grouphad 53 subjects, 32 subjects genotypes were Trp/Trp and covered 60.38%.Conclusions:1. The tolerance trial:Five dose groups in master singledosage tolerance trial had thyroid gland function abnormality; low bloodpressure;urine test abnormality; thrombocyte abnormality, but it was not toachieve the condition of stopping the trial. Two dose groups in multiple dosagetolerance trial:decreasing slightly of hemoglobin; thyroid gland ultrasonicwave and liver function abnormal, but it was not to achieve the condition ofstopping the trial too. Results primarily indicated:It's safe when the doseat 10mg-300mg, we can do the phaseâ…¡clinical trial of RS capsule at this doseand observe the same kind of adverse reaction. 2. Study on pharmacokineticsof this drug in healthy subjects:by a non-compartment model to accumulate theplasma drug concentration data. It existed in the forms of combining-type ofglycuronic acid. The food was not the factor which effects the drug. 3. The genepolymorphisms research of relative drug metabolism enzymes:the demographydistribution characteristic had no difference in the trial group andcontrols. In adverse reaction group, G/A and C/A types were more than othergenotypes. This two genotypes were maybe relevant to the adverse reaction, but we need to enlarge the sample and do research to confirm it.