In vitro metabolism of a novel antithrombotic compound,S002-333, and its enantiomers: quantitative cytochrome P450 phenotyping,metabolic profiling and enzyme kinetic studies

Abstract

1.?S002-333, (2-(4′-methoxy-benzenesulfonyl)-2,3,4,9-tetrahydro-1H-pyrido (3,4-b) indole-3-carboxylic acid amide) is a novel potent antithrombotic molecule currently under development phase. It is the racemic mixture of two enantiomers, namely S004-1032 (R-form) and S007-1558 (S-form).

2.?The contribution of five major isoenzymes, namely CYP2B6, 2C9, 2C19, 2D6 and 3A4 was quantified using recombinant P450s in the phase-I metabolism through relative activity factor approach. CYP2C19 was found to be the major contributor for S002-333 and S007-1558, while CYP3A4 showed greater involvement in S004-1032 metabolism. Chemical inhibition and immunoinhibition studies reconfirmed the results in human liver microsomes (HLM).

3.?Four major phase-I metabolites of S002-333; M-1 and M-3 (oxidative), M-2 (O-demethylated) and M-4 (dehydrogenated) were characterized in HLM. These metabolites constituted 11.2, 11.3 and 21.5% of the parent in comparison with the net phase-I metabolism of 29.9, 31.4 and 38.3% of S002-333, S004-1032 and S007-1558, respectively.

4.?Among CYP2C9, 2C19 and 3A4, the relative contribution of CYP2C9 was found to be maximum during M-1 through M-4 formation. Enzyme kinetic analysis for detected metabolites indicated that M-1 to M-3 followed classical hyperbolic kinetics, whereas M-4 showed evidence of autoactivation. In conclusion, the results suggest prominent role of CYP2C9, 2C19 and 3A4 isoforms for enantioselective disposition of S002-333 in vitro.     

A case of scirrhous gastric cancer with peritonitis carcinomatosa controlled by TS-1(R) + paclitaxel for 36 mo after diagnosis

A 34-year-old female complaining of abdominal fullness was diagnosed as scirrhous gastric cancer (type 4')with peritonitis carcinomatosa in July 2002. A combined chemotherapy regimen was selected to control massive ascites; TS-1(R) 80 mg/m2 was given orally on d 1-14,22-35, and paclitaxel 50 mg/m2 was administered intravenously on d 1, 8, 22 and 29. After 2 courses of this regimen, the primary tumor was markedly reduced,and ascites completely vanished. Alopecia (grade 1,since d 30), leukocytopenia (grade 2, on d 34) and anemia (grade 2, on d 34) were the only adverse events throughout the following courses. The chemotherapy was effective for 28 mo, and then it was discontinued upon the patient's own request, and she survived for 36mo after diagnosis.     

An oral anticancer drug, TS-1, enabled a patient with advanced gastric cancer with Virchow's metastasis to receive curative resection

We encountered a patient with advanced gastric cancer, with Virchow's lymph node metastasis, who subsequently underwent curative resection after neoadjuvant chemotherapy with the newly developed oral anticancer drug, TS-1. The patient was a 67-year-old woman who had a type 2 tumor in the middle third of the stomach, and Virchow's lymph node metastasis, which was diagnosed by fine-needle aspiration cytology; she also had swollen paraaortic lymph nodes. Curative resection was considered impossible, and TS-1 (100 mg/day) was administered for 28 days in one course, mainly in the outpatient clinic. Although grade 2 stomatitis interrupted the therapy on day 21 of the second course and on day 7 of the third course, the type 2 tumor showed marked remission (partial response; PR) and the metastasis in the Virchow's and paraaortic lymph nodes had completely disappeared after the third course (complete response; CR). Eleven weeks after the completion of the TS-1 treatment, total gastric resection with D3 lymph node dissection was performed. Histopathological examination revealed tumor involvement only in the mucosal and submucosal layers of the stomach and the no. 4d lymph node. Most of the tumor was replaced with fibrosis with granulomatous change in the muscularis propria of the stomach and in the no. 3, no. 6, and no. 7 lymph nodes. This may be the first report of a patient with advanced gastric cancer with Virchow's lymph node metastasis who successfully received curative resection following neoadjuvant chemotherapy with a single oral anticancer drug. Received: August 7, 2001 / Accepted: January 28, 2002     

Alternate-day oral therapy with TS-1 for advanced gastric cancer

Background TS-1 (1M tegafur-0.4M 5-chloro-2,4-dihydroxypyrimidine-1M potassium oxonate) has a high single-agent response rate, of more than 40%, for gastric cancer; however, the recommended regimen of 4 weeks of administration interrupted by 2 weeks of drug withdrawal frequently causes adverse effects. The alternate-day dosage of pyrimidine fluoride anticancer drugs could reduce their adverse effects without compromising their effects. We attempted an alternate-day therapy with TS-1 aiming at the avoidance of adverse effects and significantly longer duration of administration.Methods We observed patients for clinical effects and adverse effects under alternate-day dosage of TS-1, and determined blood 5-fluorouracil (FU) levels. The judgment of clinical effects was based on the New Guidelines to Evaluate the Response to Treatment in Solid Tumors (RECIST), whereas the evaluation of adverse effects was based on the National Cancer Institute NCI-common toxicity criteria (CTC).Results In 72 (78%) of 92 patients, the TS-1 regimen was converted to the alternate-day dosage because of adverse effects. Twenty patients were treated with the alternate-day dosage regimen from the start because of the fear of adverse effects. The alternate-day dosage was clinically effective, as 28 of 34 patients after relatively curative resection remained alive and free from recurrence. The median survival time of 58 patients after noncurative resection or with unresectable or recurrent cancer was 332 days. Fifty-three percent of these 58 patients achieved partial response and stable disease of more than 12 weeks duration. We followed time-dependent changes in blood 5-FU levels in 36 of the patients on alternate-day therapy, in whom TS-1 had been administered daily before being administered every other day. The trough level was significantly lower when TS-1 was administered on alternate days, and blood 5-FU reached a peak at sufficiently effective levels at 2h even after administration on the alternate-day basis.Conclusion This study demonstrated that, compared with daily administration, alternate-day administration of TS-1 reduces adverse effects, and simultaneously ensures effective blood levels and provides sufficient clinical effects.     

A Cost-Effectiveness Analysis of Organ Preservation Methods for Deceased Donor Kidneys at High Risk for Delayed Graft Function in Brazil

The clinical benefit of machine perfusion (MP) was recently assessed in a 1-year Brazilian multicenter prospective randomized trial, that showed that the use of MP was associated with a reduced incidence of delayed graft function (DGF) compared to static cold storage (SCS) in kidney transplant recipients (45% vs 61%). The objective of the present analysis is to consider the cost-effectiveness of MP relative to SCS based on clinical data from this Brazilian cohort. A decision tree model was constructed to simulate a population of 1000 kidney transplant recipients based on data derived from this Brazilian multicenter clinical trial. The model accounts for different health state utilities to estimate the cost-effectiveness of deceased donor kidney transplantation in Brazil comparing 2 kidney preservation methods: MP and SCS. The model accounts for 3 possible graft outcomes at 1 year post-transplantation: success (an immediate functioning kidney), failure (primary nonfunction requiring a return to dialysis), or DGF 1 year post-transplant. MP provided 612 total quality-adjusted life years (QALYs) (0.61 QALYs per patient) as compared to SCS (553 total QALYs, 0.55 QALYs per patient). MP was cost effective relative to SCS (US$22,117/QALY, R$70,606/QALY). The use of MP also resulted in more functioning grafts than SCS (821 vs 787), leading to a cost per functioning graft of US$38,033 (R$121,417). In conclusion, this analysis indicates that, despite the initial added cost associated with MP, the use of MP results in more functioning grafts (821 vs 787) and higher patient quality of life relative to SCS in Brazil.     

MC-002对大鼠局灶性脑缺血/再灌注治疗时间窗的研究

目的:研究受试药3-甲氧基-4-O-(2’-亚甲基-3’5’6’-三甲基吡嗪基)肉桂酸(MC-002)对大鼠局灶性脑缺血/再灌注损伤(CIRI)的治疗时间窗。方法:线栓法制作大鼠大脑中动脉闭塞(MCAO)2 h/再灌注22 h,并于复灌后0、0.5、1、2、4 h时治疗给药。观察MC-002对CIRI大鼠脑功能、脑梗死体积、脑含水量、组织形态学的影响以及MC-002各剂量组对大鼠脑组织内生化指标的影响。结果:MC-002在复灌后0、0.5、1、2 h给药能够显著性改善MCAO后大鼠的脑神经功能(P<0.05,P<0.01),降低大鼠的脑梗死率和含水量(P<0.05、P<0.01);增加CIRI大鼠脑组织超氧化物歧化酶(SOD)活力(P<0.01),降低丙二醛(MDA)及乳酸(LD)含量(P<0.05,P<0.01)。脑组织病理切片结果显示,和模型组比较,治疗组神经细胞形态和数目均得到显著改善。结论:MC-002 2 h内治疗给药对大鼠局灶性脑缺血/再灌注损伤有很好的保护作用。     

替吉奥胶囊联合紫杉醇治疗进展期胃癌的临床研究

目的探讨替吉奥胶囊联合紫杉醇治疗进展期胃癌的有效性及安全性。方法选取56例进展期胃癌患者,随机分为两组:试验组给予替吉奥胶囊80 mg.m-2.d-1,分2次口服,d1～14;紫杉醇175 mg/m2静脉滴注,d1。对照组给予表阿霉素50 mg/m2静脉滴注,d1;顺铂60 mg/m2静脉滴注,d1;5-氟尿嘧啶1 000 mg.m-2.d-1,应用静脉泵持续静脉滴注,d1～5。每21 d为1个周期,至少接受化疗2个周期。结果治疗后两组患者疗效间差异无统计学意义(P>0.05)。试验组与对照组的中位无进展生存时间分别为168 d和156 d,中位总生存时间分别为292 d和268 d,两组患者中位无进展生存时间及中位总生存时间比较,差异均无统计学意义(P>0.05)。主要毒性反应为消化道及血液学毒性,试验组消化道毒性反应比对照组轻,差异有统计学意义(P<0.05)。结论替吉奥胶囊联合紫杉醇治疗进展期胃癌安全、有效。     

LC-MS/MS快速测定大鼠血浆中HHY-002浓度

目的：建立用于测定治疗慢性心律失常药物HHY-002的大鼠血浆测定法。方法：流动相为乙腈-0．1％甲酸（65：35）,流速0．200mL／min,色谱柱Phenomenex LunaC18柱（150mm×2．00mm,5μm,5micron）,进样量为10μL,柱温35℃,以地西泮为内标。结果：HHY-002在1．98～1013．50ng／mL的范围内呈良好的线性关系（r=0．998）,最低定量限达1．98ng／mL,绝对回收率高于80％,日内和日间误差小于10％,方法回收率大于（81．4±4．5）％,符合生物样品分析要求。结论：建立的LC—MS／MS方法专属性强,灵敏度高,可用于HHY-002的体内定量分析。     

RI-002, an intravenous immunoglobulin containing high titer neutralizing antibody to RSV and other respiratory viruses for use in primary immunodeficiency disease and other immune compromised populations

Introduction: Novel immune globulin (IG) products (RI-002, RI-001) have been designed to provide protection against respiratory syncytial virus (RSV) mediated respiratory illness while at the same time meeting the manufacturing requirements established by FDA for antibody supplementation in immunocompromised subjects.

Areas covered: This review covers the manufacture and development of both RI-001 and RI-002, including the selection of plasma donors for IG preparation with high-titers of anti-RSV antibody, in vitro, and preclinical data in the cotton rat model S. hispidus, and clinical trials including Phase II and compassionate use studies of RI-001 and a multi-center, pivotal Phase III study of RI-002 in PIDD patients.

Expert commentary: The data demonstrate that RI-002 is efficacious in the prevention and treatment of RSV in preclinical normal and immune suppressed animal models and is safe and efficacious in the treatment of patients with various forms of primary immunodeficiency disease (PIDD). This product offers potential advantages over other available IG’s for prophylaxis in immunocompromised patients requiring polyclonal immunoglobulin supplementation because of its unique antibody composition. In addition to its enhanced neutralizing anti-RSV activity and its polyclonal IG composition, there is preclinical data to support the use of RI-002 for humoral protection against other respiratory pathogens.