新HIV融合抑制剂西夫韦肽的临床药代动力学及其与生物膜作用机理研究

发布时间：2018-06-26 08:02

本文选题：西夫韦肽 + 临床药代动力学　；参考：《北京理工大学》2014年博士论文

【摘要】：西夫韦肽是基于HIV-1膜融合蛋白gp41的核心构象的三维结构全新设计开发的新一代HIV融合抑制剂,具有良好的抗HIV药物开发前景。为了对一类新抗HIV药物西夫韦肽的临床药代动力学、分子水平的药理机制做出全面深入评价与研究,本文首先建立了HIV感染者体内西夫韦肽的液质联用定量方法,并对方法进行了完整详细的确证性考察,结果表明液质联用定量方法灵敏度高达9.75 ng/mL;批内(间)准确度在-8.37~2.53%之间,精密度CV在2.74~7.57%之间;回收率在75.60~80.35%之间;血浆样品在各种不同贮存条件下,准确度在-6.56~4.51%之间,精密度CV在3.86~10.41%之间,表明各项方法学指标考察结果均符合新药临床药代动力学研究要求,适用于HIV感染者血浆中西夫韦肽的含量测定。本文利用所建方法对IIa临床试验两个剂量组(10 mg、20 mg)共19名无治疗经验的HIV感染者每天一次皮下注射西夫韦肽连续给药28天的药代动力学进行了研究。结果表明在两个剂量组(10 mg、20 mg)的病人体内,西夫韦肽被迅速吸收,达峰时间短,表观分布较广,半衰期长,表现出良好的药代动力学行为。连续28天多次给药两个剂量组病人血药浓度平稳;累积比达到1.7和1.3,表现体内有一定蓄积现象。在IIb临床试验中,对8名有治疗经历的HIV感染者皮下注射西夫韦肽联合口服HAART(HighActiveAnti-Retrovirul Therapy)药物连续治疗168天的药代动力学进行了研究。结果显示HIV感染者连续128天给予20 mg西夫韦肽联合HAART治疗的药时曲线表明平均波谷浓度Cmin均远大于体外药效试验EC50;首次给药后及末次给药后各药代动力学参数均无显著统计意义差异(P0.05),说明168次连续给药后,首末次给药的药代动力学情况无显著变化;联合用药病人代谢明显加快,表现为清除速率加快,消除半衰期变短,西夫韦肽的药代动力学可能受到与HAART药物合用的影响。本文第二部分首先建立了基于表面等离子体共振(SPR)的多肽-单双层生物膜相互作用研究系统,通过数据拟和结合模型分析,研究西夫韦肽(及恩夫韦肽)与生物膜亲和常数的定量信息、多肽与生物膜结合的相互作用力的性质(疏水作用及静电作用)、多肽与生物膜结合方式(表面接合及插膜、孔道)的定性信息,以及动力学过程信息(一步结合、两步法反应结合),从分子水平完善了西夫韦肽融合抑制的药理机制。结果表明西夫韦肽首先在疏水相互作用或静电吸附的驱动下,以两步反应的方式,选择性吸附和富集在生物膜受体丰富的刚性区域,只是膜表面结合,未能插膜或形成孔道,再与gp41结合发挥融合抑制药效。而恩夫韦肽对所有磷脂膜均有一定程度的结合,并不特异选择性吸附于饱和程度较高的磷脂构成的单(双)层生物膜,表明恩夫韦肽的磷脂膜普遍结合能力是其发挥融合抑制药效的途径。两者对于生物膜选择性的不同既表明两者分子水平上不同的药理机制,也解释了西夫韦肽药效强于恩夫韦肽的原因。本研究首次对无治疗经历以及有HAART治疗经历的HIV感染者中注射用西夫韦肽药代动力学以及安全性、疗效等的长期跟踪和完整评价报道,直接为西夫韦肽的药物上市审批提供第一手的临床依据,为不同治疗经历的HIV感染者西夫韦肽的单用和与HAART药物联用的临床给药方案的完善提供了依据。第二部分的创新性在于建立了新型基于SPR的多肽-单双层生物膜相互作用研究系统,并首次应用于多肽类融合抑制剂-生物膜相互作用研究,克服了目前其他方法不能解析多肽生物膜相互作用实时信息的缺陷,首次完整阐明了西夫韦肽和恩夫韦肽分子水平上与生物膜相互作用的膜向性、作用力分析、动力学过程,并对西夫韦肽、恩夫韦肽已知融合抑制的药理机制做出了进一步完善,对西夫韦肽药效远好于恩夫韦肽做出了合理解释,为今后多肽类药物与生物膜相互作用研究提供了新的方法选择,也为HIV融合抑制剂筛选靶标提供了新的思路。
[Abstract]:Siv Vee is a new generation of HIV fusion inhibitor based on the core conformation of the core conformation of HIV-1 membrane fusion protein gp41. It has a good prospect of anti HIV drug development. In order to make a comprehensive evaluation and Research on the clinical pharmacokinetics and molecular mechanism of a class of new anti HIV drugs, the molecular level mechanism of molecular level is thoroughly evaluated and studied. First, a quantitative method of liquid mass combined with SF in HIV infected people was established and a complete and detailed investigation was carried out. The results showed that the sensitivity of the quantitative method was up to 9.75 ng/mL; the intra (inter) accuracy was between -8.37~2.53% and CV in 2.74~7.57%; the recovery rate was between 75.60~80.35% and plasma samples. Under various storage conditions, the accuracy is between -6.56~4.51% and the precision of CV is between 3.86~10.41%. It shows that all the results of the methodology are in line with the requirements of the study of the pharmacokinetics of the new drug. It is suitable for the determination of the content of the plasma in the plasma of the HIV infected people. We use the method built for the two dose groups of the IIa clinical trial. (10 mg, 20 mg) a total of 19 HIV infected people with no therapeutic experience were studied for 28 days by subcutaneous injection of the subcutaneous injection of Western Fu. The results showed that in two dose groups (10 mg, 20 mg), Western FV was quickly absorbed, the peak time was short, the apparent distribution was wide, the half-life was long, and a good drug generation was shown. Dynamic behavior. The blood concentration of patients with two doses of medication for 28 consecutive days for 28 consecutive days was stable; the cumulative ratio reached 1.7 and 1.3, showing a certain accumulation in the body. In the IIb clinical trial, 8 patients with therapeutic HIV infection were subcutaneously injected with Western Fu Wei peptide combined with oral HAART (HighActiveAnti-Retrovirul Therapy) for 168 days. The results of pharmacokinetics were studied. The results showed that the curves of HIV infected people with 20 mg and HAART for 128 days showed that the average trough concentration Cmin was far greater than that of EC50 in vitro, and there was no statistically significant difference in the pharmacokinetic parameters after and after the first administration (P0.05), indicating 168 consecutive times. In the second part, the second part first established a surface plasmon resonance (SPR). The second part of the article first established a surface plasmon resonance (SPR). The research system of the interaction of polypeptides and double layers biofilm. Through the analysis of data quasi and binding models, the quantitative information of the affinity constants of the peptide and the biofilm, the properties of the interaction between the peptide and the biofilm (hydrophobic interaction and electrostatic action), the binding of the peptide and the biofilm (surface conjugation and intercalation, hole) The qualitative information of the road, as well as the dynamic process information (one step combined with the combination of the two step method), improved the pharmacological mechanism of the fusion inhibition from the molecular level. The results showed that the SFG first was selectively adsorbed and enriched in the biofilm receptor, driven by hydrophobic interaction or electrostatic adsorption. The rich rigid area is only the membrane surface binding, failure to insert the membrane or form the channel, and then combine with the gp41 to inhibit the efficacy of fusion, while Don J Vee has a certain combination of all phospholipid membranes, which is not selectively adsorbed on the single (double) layer of high saturated phospholipids, indicating that the phospholipid membrane of the peptide is generally nodding. Combining ability is the way to play the inhibitory effect of fusion. The difference in selectivity to the biofilm shows both the different pharmacological mechanisms at the molecular level and the reason for the stronger efficacy of the drug than emweinin. This study was the first time for the non treatment experience and the injection of the Western Fu Wei peptide drug in the patients with HIV infection with HAART therapy. The long-term tracking and complete evaluation of the pharmacokinetics, safety, efficacy, and so on provide the first hand clinical basis for the approval of the drug listing for the drugs of the HIV, which provides the basis for the improvement of the single use of the HIV infected people with different treatment experiences and the clinical drug delivery scheme combined with HAART drugs. The second part is innovative. A new SPR based polypeptides single layer biomembrane interaction study system was established, and it was first applied to the study of polypeptides fusion inhibitors biological membrane interaction, which overcome the shortcomings of other methods that can not analyze the real-time information of the interaction of polypeptides, and first fully elucidated the water of Siv Vee and Don J Vee molecules. The film direction, force analysis and kinetic process of the interaction with the biofilm have been further improved, and the pharmacological mechanism of the known fusion inhibition of Siv Vee and En Fuwei peptide has been further perfected, and a reasonable explanation for the efficacy of the Western husband peptide is better than that of the weilweir peptide, which provides a new study for the study of the interaction of polypeptide drugs and biofilm in the future. The method also provides a new idea for screening targets of HIV fusion inhibitors.
【学位授予单位】：北京理工大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R969.1

【共引文献】