Velpatasvir   中文名称: 维帕他韦

HCV/hepatitis C virus nonstructural protein 5A (NS5A)

体外活性：Velpatasvir（也称为 GS-5816）是一种新型丙型肝炎病毒（HCV）非结构蛋白 5A（NS5A）的泛基因型抑制剂，具有对抗基因型 1（GT1）至 GT6 HCV 复制子的活性。它是 HCV RNA 复制的选择性抑制剂，对 GT1 至 GT6 的平均 50% 有效浓度 (EC50) 为 6 至 130 pM。激酶测定：Velpatasvir (VEL，GS-5816) 是一种新型泛基因型丙型肝炎病毒 (HCV) 非结构蛋白 5A (NS5A) 抑制剂，具有对抗基因型 1 (GT1) 至 GT6 HCV 复制子的活性。细胞分析：Velpatasvir (VEL, GS-5816) 是一种新型泛基因型丙型肝炎病毒 (HCV) 非结构蛋白 5A (NS5A) 抑制剂，具有对抗基因型 1 (GT1) 至 GT6 HCV 复制子的活性。在一项 1b 期 3 天单药治疗研究中，接受 150 mg 剂量 GS-5816 治疗的患者 GT1a、-1b、-2、-3 和 - 的平均最大 HCV RNA 下降≥3.3 log10 IU/ml 4.该报告描述了这些患者对 VEL 的病毒学耐药性。在 22/70 名患者的预处理中，通过深度测序（1% 截止值）检测到 NS5A 耐药相关替代 (RAS)，即 10/35 (29%) 患者患有 GT1a，1/8 (13%) 患有 GT1b，4/ GT2 为 8（50.0%），GT3 为 5/17（29.4%），GT4 为 2/2（100.0%）。在 GT1a 和 GT3 患者中，与没有预处理 RAS 的患者相比，预处理 RAS 与 HCV RNA 反应略有降低有关；在 GT1b、GT2 和 GT4 患者中，治疗前有或没有 RAS 的患者的反应没有观察到显着差异。治疗后，GT1a 出现的 RAS 模式比其他基因型更复杂。在 GT1a 中，取代出现在位置 M28、Q30、L31、P32、H58、E92 和 Y93，其中最常见的取代出现在位置 Y93、M28 和 L31。在GT1b和GT2中的两个位置（Y93和L31）、GT3中的三个位置（Y93、L31和E92）以及GT4中的四个位置（L28、M31、P32L和Y93）观察到RAS。治疗前存在的 RAS 在 48 周的随访期间持续存在；然而，在治疗期间出现的 RAS 在随访期间更有可能在病毒群体中的患病率和频率上下降。 （本研究已在 ClinicalTrials.gov 注册，注册号为 NCT01740791。）。

泛基因型[1]
在ASTRAL的几项研究中，Feld等人进行了一项为期12周的3期双盲安慰剂对照研究，涉及来自全球的624名慢性HCV患者，代表基因型1、2、4、5和6。患者被随机分配到治疗组（每天一次400 mg sofosbuvir和100 mgVelpatasvir的联合片剂）或安慰剂组。在治疗后12周，测量HCV RNA水平以评估SVR-12率高于85%所定义的治疗效果。分配到治疗组的患者概况包括代偿性肝硬化（19%）、既往治疗经验（32%），其中主要包括聚乙二醇干扰素治疗（89%），平均年龄为54岁。Feld等人报告称，所有患者群体的SVR率都很高，无论是治疗新手还是有经验的患者。虽然分配到安慰剂组的患者没有引起SVR，但分配到索非布韦/维帕他韦联合治疗的患者有99%的SVR。该药物组合对所有HCV基因型均有效，基因型2、4和6的SVR率为100%，基因型1b为99%，基因型1a为98%，基因型5为97%。此外，无论患者是否患有肝硬化以及是否接受过既往治疗，SVR率仍然很高。尽管主要目的是评估疗效，但治疗安全性是通过不良反应率和患者报告的结果来记录的。只有2名接受治疗的患者（均感染基因型1）出现了病毒学失败，而大约2%的患者出现了严重的不良反应。此外，索非布韦/维帕他韦治疗不会增加不良健康事件的易感性。 难以治愈的人群[1]
为了解决可能难以治愈的人群，Foster等人与ASTRAL-2和ASTRAL-3研究人员一起进行了两项随机、3期、开放标签研究。他们招募了有和没有治疗经验、有和没有肝硬化的基因型2或3患者。根据基因型将患者分开，并随机分配到两个治疗组：每天一次的400 mg索非布韦和100 mg维帕他韦联合片，持续12周，或400 mg索非布韦加基于体重的利巴韦林，持续12或24周。sofosbuvir/Velpatasvir联合片剂治疗组中基因型2的患者实现了99%的SVR。索非布韦加基于体重的利巴韦林联合治疗12周是基因型2 HCV患者的替代方案，无肝硬化，94%的病例达到SVR。尽管该组患者的治疗时间延长了24周，但这种相同的方案可以应用于SVR达到80%且耐受性相似的基因型3 HCV患者。在ASTRA-3试验中，基因型3治疗初治和有经验的患者被分为两个治疗组，一组每天接受12周的索非布韦/维拉帕斯韦治疗，另一组接受24周的索非布韦加基于体重的利巴韦林治疗。在索非布韦/维拉斯韦治疗组中，95%的基因型3患者实现了SVR（95%CI，92%-98%），而索非布维和基于体重的抗病毒治疗组的SVR实现率为80%（CI，75%-85%）。

在sofosbuvir/Velpatasvir组中，肝硬化患者的疗效也得到了评估，他们的SVR达到了91%，而sofosbuvir/利巴韦林组的SVR为66%。索非布韦/利巴韦林组的不良健康事件发生率（表1）高于索非布韦/维帕他韦组（71.3%比52.3%）。目前，对于慢性肾病或肌酐清除率低于30 mL/min的患者，索非布韦不需要调整剂量。与ASTRAL-1研究的情况一样，Younossi等人记录了治疗期间和治疗后患者报告的结果。治疗四周后，在sofosbuvir/velatasvir治疗的患者中，许多前域都有统计学和临床上的显著改善，包括身体和情绪健康、身体疼痛和一般健康。索非布韦/利巴韦林治疗产生了不同的结果，患者报告身体疼痛加剧，情绪健康和幸福感较差。患者还报告社会功能和身体健康受损，降至基线水平以下。索非布韦/维帕他韦对全身健康，特别是难以治愈的基因型3患者的直接益处优于索非布韦加基于体重的利巴韦林治疗。

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GT1至GT4 HCV感染患者对Velpatasvir/VEL的抗病毒反应。[2]
VEL单药治疗三天后，HCV RNA水平迅速下降。在服用150mg VEL的队列中，GT1a、GT1b、GT2、GT3和GT4组在第17天的HCV RNA水平中位数分别为4.19、4.29、4.39、3.13和3.17 log10 HCV RNA IU/ml（表1）。服用5、25、50或100 mg的GT1a患者的HCV RNA中位数减少>3.67 log10，服用25或50 mg的GT3患者的中位数减少>3.12 log10。

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NS5A-RAS的作用存在预处理。[2]
对于所有接受Velpatasvir/VEL治疗的70名患者和8名安慰剂治疗的患者，分析了氨基酸28、30、31、32、58、92和93处已知NS5A抑制剂RAS多态性的序列。24名患者在治疗前存在NS5A RAS（检测率>1%），即2/8名安慰剂、10/35名GT1a、1/8名GT1b、4/8名GT2、5/17名GT3和2/2名GT4患者（表1），其中一些患者的RAS>1。接受VEL治疗的7名患者在氨基酸残基93（Y93C/F/H/N）处存在治疗前RAS（表2）。对于GT1a，在M28T、Q30H/K/R、L31M/V、H58D和Y93C/H/F/N位置观察到NS5A RAS。服用150mg VEL并接受预处理NS5A RAS的GT1a患者的平均病毒载量减少为2.9 log10（表1），而未接受预处理的NS5A RAS患者的平均减少量为4.38 log10（见表1）。接受NS5A RAS预处理（Y93H）的GT1b患者的HCV RNA减少量为4.47 log10，而未服用NS5A RASs的患者的平均降低量为4.39 log10（见图1）。在4名接受L31M预处理的GT2患者中，log10平均降低4.08 log10，而未接受NS5A RAS的患者为4.62 log10（表1）。在5名接受NS5A RAS预处理的GT3患者中，2名接受25mg VEL治疗，1名接受50mg VEL治疗。接受25或50mg VEL治疗的治疗前RAS的GT3患者HCV RNA平均减少<1log10，而这些剂量组中未接受治疗前RAS治疗的所有GT3患者的HCV RNA减少>3-log10。两名接受150mg VEL治疗的RAS患者的HCV RNA平均减少2.9 log10和2.7 log10，而没有NS5A RAS的患者为3.54 log10（表1；图1）。在30位有变异的GT4患者（一名为L30H[45.8%]和L30R[53.7%]，一名为L3 0S[2.4%]和L3 0H[97.1%]）的HCV RNA平均减少3.47 log10。

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在接受Velpatasvir/VEL治疗至第17天的患者HCV中选择替代品。[2]
为了鉴定可能与VEL病毒学耐药性相关的HCV NS5A变异，在治疗期间和治疗后通过深度测序以1%的截止值分析了全长NS5A编码区。分别在治疗期间（第2至3天）或治疗后（第4至10天）和第17天获得样本，并在病毒载量≥1000 IU/ml时进行分析。在没有预处理NS5A RAS的46/70名VEL治疗患者中，分别在第2至10天和第17天可获得40和46的序列。所有在第2至10天具有可用序列的患者（40/40；100%）都出现了紧急NS5A RAS，80.4%（37/46）在第17天仍有RAS（表3）。在对样本进行测序的两名安慰剂治疗患者的治疗后期间，未检测到出现的NS5A RAS。与其他GT患者相比，GT1a患者在治疗中出现的NS5A RAS位置更多，包括M28、Q30、L31、P32、H58、E92和Y93位置的置换。位置Y93、M28和L31的RAS在GT1a患者中最为常见（表4）。在GT1b和GT2患者（Y93、L31）的两个NS5A位置和GT3患者（Y1993、L31、E92）的三个位置观察到RAS。在两名GT4患者中，NS5A RAS出现在L28、M31、P32L和Y93位置（表5）。L31M/V和Y93H是GT1b和GT2患者在治疗中最常见的RAS，E92K和Y93H/N是GT3患者中最普遍的RAS（表4）。

病毒测序。[2]
对于每一名接受 Velpatasvir/VEL治疗的患者和17名接受安慰剂治疗的患者中的8名，在预处理访视和第4天和/或第2、4、5、7、10或17天以及随访第12、24和48周，使用HCV RNA水平>1000 IU/ml的样本扩增HCV NS5A基因，使用Illumina MiSeq平台（加利福尼亚州圣地亚哥Illumina）以1%的检测灵敏度截止值对其进行深度测序，但1名在预处理随访时进行人群测序的患者除外。Janssen Diagnostics（比利时比尔斯）通过逆转录PCR和批量PCR产物的标准Sanger测序对全长HCV NS5A编码区进行了群体测序。检测抗性变体的灵敏度约为10%至20%。变异被报告为与基因型特异性参考菌株的差异，即GT1b Con1（AJ238799）、GT1a H77（GenBank登录号NC_004102）、GT2 JFH-1（AB047639）、GT3 S52（GU814263）或GT4 ED43（GU8142465）。通过多步方法，使用内部开发的软件对深度测序读数进行比对和处理，以识别含量>1%的替代物。测序分析包括HCV耐药性咨询小组总结的NS5A类RAS和/或最近在LDV、Velpatasvir/VEL、DCV、ABT-267、ABT-530和MK-8742的临床试验中观察到的RAS，包括位置24、28、30、31、32、38、58、92和93。

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复制子RNA瞬时转染Huh7细胞并测定EC50。[2]
通过定点突变将抗性突变引入GT1a、GT1b、GT2a、GT3a和GT4a复制子（分别为骨架GT1a H77、GT1b Con1、GT2a JFH-1、GT3a S52和GT4a ED43），并如前所述在瞬时转染中进行测试。简而言之，根据制造商的说明，使用QuikChange II XL诱变试剂盒将NS5A突变引入编码PI-hRluc复制子的质粒中。DNA测序证实了突变。使用MEGAscript试剂盒从复制子编码质粒中体外转录复制子RNA。通过Lohmann等人的方法将RNA转染到Huh-lunet细胞中。简而言之，细胞被胰蛋白酶消化，并用磷酸缓冲盐水（PBS）洗涤两次。将4×106个细胞在400μl PBS中的悬浮液与5μg RNA混合，在960μF和270V的设置下进行电穿孔。将细胞转移到40ml预热的培养基中，然后接种到96孔板（100μl/孔）中。将化合物在100%二甲亚砜（DMSO）中连续稀释3倍，并以1:200的稀释度加入细胞中，使DMSO的最终浓度为0.5%，总体积为200μl/孔。将细胞处理3天，之后移除培养基，裂解细胞，并用市售的测定法和Top Count仪器定量Renilla萤光素酶活性。EC50计算为与DMSO对照样品相比，观察到Renilla报告活性水平降低50%的化合物浓度。通过非线性回归分析，使用GraphPad Prism软件包生成剂量反应曲线和EC50s。参考菌株（1b-Con1和1a-H77）或从临床分离株瞬时衍生的嵌合复制子的复制水平被确定为电穿孔后第4天的肾荧光素酶信号与4小时的信号之比，以使转染效率正常化。在同一实验中，每个复制子的复制能力表示为与参考菌株（1b-Con1或1a-H77）相比的归一化复制效率。

Velpatasvir (VEL, GS-5816) 是一种新型泛基因型丙型肝炎病毒 (HCV) 非结构蛋白 5A (NS5A) 抑制剂，具有对抗基因型 1 (GT1) 至 GT6 HCV 复制子的活性。在一项 1b 期 3 天单药治疗研究中，接受 150 mg 剂量 GS-5816 治疗的患者 GT1a、-1b、-2、-3 和 - 的平均最大 HCV RNA 下降≥3.3 log10 IU/ml 4.该报告描述了这些患者对 VEL 的病毒学耐药性。在 22/70 名患者的预处理中，通过深度测序（1% 截止值）检测到 NS5A 耐药相关替代 (RAS)，即 10/35 (29%) 患者患有 GT1a，1/8 (13%) 患有 GT1b，4/ GT2 为 8（50.0%），GT3 为 5/17（29.4%），GT4 为 2/2（100.0%）。在 GT1a 和 GT3 患者中，与没有预处理 RAS 的患者相比，预处理 RAS 与 HCV RNA 反应略有降低有关；在 GT1b、GT2 和 GT4 患者中，治疗前有或没有 RAS 的患者的反应没有观察到显着差异。治疗后，GT1a 出现的 RAS 模式比其他基因型更复杂。在 GT1a 中，取代出现在位置 M28、Q30、L31、P32、H58、E92 和 Y93，其中最常见的取代出现在位置 Y93、M28 和 L31。在GT1b和GT2中的两个位置（Y93和L31）、GT3中的三个位置（Y93、L31和E92）以及GT4中的四个位置（L28、M31、P32L和Y93）观察到RAS。治疗前存在的 RAS 在 48 周的随访期间持续存在；然而，在治疗期间出现的 RAS 在随访期间更有可能在病毒群体中的患病率和频率上下降。

Clinical trial population and study design.[2]
This was a phase 1b double-blind, randomized, placebo-controlled, multicenter study of Velpatasvir/VEL in HCV-infected patients in the United States (ClinicalTrials.gov identifier NCT01740791). Clinical data of this trial have been described previously. A total of 87 patients were enrolled and received treatment in 1 of 11 cohorts, each randomized 4:1 to treatment with VEL or a matching placebo for 3 days (except for the GT4 patients, who were not randomized and received VEL). The actual treatments administered are presented in Table 1. One patient discontinued study treatment because of an adverse event, and two patients discontinued the study (one was lost to follow-up, and one withdrew consent) prior to day 17 assessments (these patients were included in the sequencing analyses). VEL was administered once daily as follows: 5, 25, 50, 100, and 150 mg to GT1a patients; 150 mg to GT1b, GT2, and GT4 patients; and 25, 50, and 150 mg to GT3 patients. Eligible patients had plasma HCV RNA levels of >5 log10 IU/ml pretreatment and were treatment naive. Of the 87 patients in this study, 45 had HCV GT1a, 10 had GT1b, 10 had GT2b, 1 had GT3, 19 had GT3a, 1 had GT4, and 1 had GT4a, GT4b, and GT4c. The study was conducted in compliance with the Declaration of Helsinki. The study protocol and informed consent documents were reviewed and approved by the institutional review board of the participating institution, and informed consent was obtained from all patients before any study-specified procedures.

Absorption, Distribution and Excretion
Oral bioavailability is 25-30%. 94% is excreted in feces, of which 77% is the unchanged drug. 0.4% is excreted in urine. 1.4-1.6 L/kg. Estimated absorption rate is 0.12 L/h/kg [A19175]. Metabolism/Metabolites Partially metabolized by CYP2B6, CYP2C8, and CYP3A4. Biological Half-Life 15 hours.

Effects During Pregnancy and Lactation
◉ Summary of Lactation Use
No studies have been conducted on velpatasvir in breastfeeding women undergoing treatment for hepatitis C. Because it binds to maternal plasma proteins at a rate exceeding 99%, its concentration in breast milk is likely to be very low. Some sources suggest that breastfeeding should be avoided when velpatasvir is used in combination with ribavirin.
Hepatitis C is not transmitted through breast milk, and breast milk has been shown to inactivate the hepatitis C virus (HCV). However, the U.S. Centers for Disease Control and Prevention (CDC) recommends that breastfeeding should be considered if the mother has cracked or bleeding nipples. It is unclear whether this warning applies to mothers undergoing treatment for hepatitis C.
Infants born to HCV-infected mothers should be tested for HCV infection; nucleic acid testing is recommended because maternal antibodies are present in the infant for the first 18 months after birth and before the infant develops an immune response.
◉ Impact on Breastfed Infants
No published information was found as of the revision date.
◉ Effects on lactation and breast milk
No relevant published information was found as of the revision date.

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Protein binding
>99.5% bound to plasma proteins.

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Drug interactions[1]
An open-label study evaluated potential drug interactions between sofosbuvir and velpatasvir. The study included 18 healthy individuals without HCV infection. No clinically significant drug interactions were reported in the study by Mogalian et al. When using sofosbuvir or a sofosbuvir-containing regimen, patients should be advised to avoid rifampin, St. John's wort, or tepranavir, as these drugs can reduce circulating drug concentrations of sofosbuvir, thereby reducing its efficacy (Table 2). Velpatasvir absorption may be reduced if patients are taking antacids and acid-suppressing drugs concurrently. Patients should be informed of all medications they are currently taking and whether they plan to start any other medications, including over-the-counter drugs or herbal supplements.

[1]. Drug Des Devel Ther.2017 Feb 23;11:497-502.

[2]. Antimicrob Agents Chemother.2016 Aug 22;60(9):5368-78.

[3]. Signal Transduct Target Ther. 2021 May 29;6(1):212.

Pharmacodynamics
Velpatasvir is a small-molecule direct-acting antiviral drug used in combination with sofosbuvir to treat hepatitis C. Velpatasvir inhibits viral replication by suppressing non-structural protein 5A (NS5A). Even at doses up to five times the recommended dose, velpatasvir does not prolong the QTc interval to any clinically significant extent. Velpatasvir is a complex organic heteropentacyclic compound and an inhibitor of hepatitis C virus non-structural protein 5A, used in combination with sofosbuvir (trade name Epclusa) to treat patients with chronic hepatitis C of all six major genotypes. It is both an antiviral drug and an inhibitor of hepatitis C virus non-structural protein 5A. It is an organic heteropentacyclic compound belonging to the classes N-acylpyrrolidine, L-valine derivatives, carbamates, imidazoles, cyclic compounds, and ethers. Velpatasvir is a direct-acting antiviral (DAA) used in combination therapy for chronic hepatitis C. Chronic hepatitis C is an infectious liver disease caused by hepatitis C virus (HCV) infection. HCV is a single-stranded RNA virus with nine different genotypes, of which genotype 1 is the most common in the United States, affecting 72% of chronic HCV patients. Velpatasvir is a defective substrate of NS5A (non-structural protein 5A), a non-enzymatic viral protein that plays a crucial role in the replication, assembly, and regulation of the host immune response of hepatitis C virus. Since 2011, significant progress has been made in the treatment options for chronic hepatitis C with the development of direct-acting antiviral agents (DAAs) such as velpatasvir. Notably, velpatasvir has a significantly higher resistance barrier than first-generation NS5A inhibitors, such as [DB09027] and [DB09102], making it a highly effective and reliable alternative therapy for chronic hepatitis C. In a joint guideline published in 2016, the American Association for the Study of Liver Diseases (AASLD) and the Infectious Diseases Society of America (IDSA) recommended velpatasvir in combination with sofosbuvir as a first-line treatment for all six genotypes of hepatitis C virus. Currently, velpatasvir is only marketed as the combination drug Epclusa, which contains velpatasvir and another direct-acting antiviral agent [DB08934]. Treatment goals for Epclusa include cure or achieving sustained virological response (SVR), defined as achieving this response after 12 weeks of daily treatment. SVR and eradication of HCV infection are associated with significant long-term health benefits, including reduced liver-related damage, improved quality of life, reduced incidence of hepatocellular carcinoma, and reduced all-cause mortality and liver transplantation risk. Since June 2016, velpatasvir and [DB08934] have been marketed as a fixed-dose combination drug under the brand name Epclusa. Epclusa is the first approved combination therapy for hepatitis C (HCV) of all genotypes (with or without cirrhosis). It is also currently the most potent HCV antiviral drug on the market, achieving a sustained virological response (SVR) rate of 93-99% after 12 weeks of treatment, depending on genotype and degree of cirrhosis, and exhibiting a high resistance barrier. Guidelines in Canada and the United States recommend Epclusa as a first-line treatment for all HCV genotypes. Velpatasvir is a hepatitis C virus NS5A inhibitor. Velpatasvir's mechanism of action includes inhibition of breast cancer resistance protein, P-glycoprotein, organic anion transport peptide 1B1, organic anion transport peptide 1B3, and organic anion transport peptide 2B1. Velpatasvir is an oral inhibitor of the hepatitis C virus (HCV) nonstructural protein 5A (NS5A) replication complex, with potential activity against HCV genotypes 1-6. Although the exact mechanism of action of velpatasvir is not fully elucidated, it appears to bind to domain I of the NS5A protein after oral administration and cellular uptake. This inhibits NS5A protein activity, leading to the disruption of the viral RNA replication complex, blocking HCV viral RNA production, and inhibiting viral replication. NS5A is a proline-rich, zinc-binding, hydrophilic phosphoprotein that plays a crucial role in HCV RNA replication. HCV is a small, enveloped, single-stranded RNA virus belonging to the Flaviviridae family. VELPATASVIR is a small molecule drug, currently in Phase IV clinical trials (covering all indications), and was first approved in 2016 for the treatment of chronic hepatitis C virus infection, with one investigational indication. It is an open target. Hepatitis C virus (HCV) is a global epidemic, infecting nearly 200 million people worldwide. HCV is the most common blood-borne infection in the United States and can lead to a variety of health problems, including liver fibrosis, cirrhosis, and hepatocellular carcinoma. Traditional genotype-based hepatitis C virus (HCV) treatment regimens, such as interferon, have achieved some success in sustained clearance of the viral genome. A recent clinical trial showed that a once-daily combination of sofosbuvir (a non-structural protein 5B polymerase inhibitor) and velpatasvir (a non-structural protein 5A inhibitor) achieved a sustained virological response rate of approximately 95% in all HCV genotypes, regardless of prior treatment history or presence of cirrhosis. Patients reported improvements in overall health, fatigue, mood, and mental health after completing the combination therapy. This combination therapy is highly effective, but should be used with caution in patients taking certain medications or with certain diseases. This article will review the safety and efficacy of the sofosbuvir/velpatasvir combination regimen for all HCV genotypes. [1]

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The advent of direct-acting antiviral agents (DAAs), designed to selectively target the HCV replication process, has spurred a great deal of scientific research and has yielded encouraging results in various aspects. To date, the potent combination therapy of sofosbuvir (an NS5B protease inhibitor) and velpatasvir (an NS5A inhibitor) has demonstrated sustained virological response (SVR) rates exceeding 94% in all major HCV genotypes, including treatment-resistant genotypes, patients with cirrhosis, and previously treated patients (Table 3). This regimen holds promise for virological cure, as SVR rates of 95% or higher have been reported. Furthermore, a significant proportion of patients have experienced improvements in overall health, mood, mental well-being, and social productivity during and after treatment, further demonstrating the effectiveness of this combination therapy. Currently, insurance companies determine HCV treatment eligibility based on pre-treatment genotyping and liver fibrosis staging. While the sofosbuvir/velpatasvir combination therapy has been successful in all genotypes, offering a simple, safe, and potentially curative treatment option, its accessibility remains limited by cost. Once all patients have access to treatment, this will reduce the healthcare burden, thereby lowering morbidity and mortality, and patients will be able to be cured of hepatitis C virus infection without genotyping. [1] Velpatasvir (VEL, GS-5816) is a novel pangenotypic hepatitis C virus (HCV) nonstructural protein 5A (NS5A) inhibitor that is active against HCV replicons from genotype 1 (GT1) to GT6. In a 3-day phase 1b monotherapy study, patients treated with 150 mg of GS-5816 showed a mean maximum decrease in HCV RNA for GT1a, -1b, -2, -3, and -4 genotypes ≥3.3 log10 IU/ml. This report describes the virological resistance to VEL in these patients. Of the 70 patients, 22 had NS5A resistance-associated mutations (RAS) detected by deep sequencing (1% threshold) before treatment. These included 10 out of 35 GT1a patients (29%), 1 out of 8 GT1b patients (13%), 4 out of 8 GT2 patients (50.0%), 5 out of 17 GT3 patients (29.4%), and 2 out of 2 GT4 patients (100.0%). In GT1a and GT3 patients, those with pre-treatment RAS showed slightly lower HCV RNA responses than those without. However, in GT1b, GT2, and GT4 patients, there was no significant difference in response between those with and without pre-treatment RAS. Post-treatment, the newly emerging RAS pattern in GT1a patients was more complex than in other genotypes. In GT1a, amino acid substitutions were observed at M28, Q30, L31, P32, H58, E92, and Y93 sites, with substitutions at Y93, M28, and L31 being the most common. In GT1b and GT2, resistance-associated mutations (RAS) were observed at Y93 and L31 sites; in GT3, resistance-associated mutations were observed at Y93, L31, and E92 sites; and in GT4, resistance-associated mutations were observed at L28, M31, P32L, and Y93 sites. Pre-treatment resistance-associated mutations persisted during the 48-week follow-up period; however, resistance-associated mutations that appeared during treatment were more likely to decrease in prevalence and frequency in the viral population during the follow-up period. (This study has been registered at ClinicalTrials.gov, registration number NCT01740791.) [2] Phenotypic analysis of RAS detected before treatment or screened after velpatasvir/VEL treatment showed that most RAS in the GT1a replicon did not show resistance (EC50 change ≤ 2.5-fold, Q30L/R/H, Y93F) or low to moderate levels of resistance (EC50 change 2.5 to 100-fold, Q30K/E, L31I/M/V, P32L, H58D, Y93C/S). High levels of resistance (EC50 change > 100-fold) were observed in the Y93H/N/R/W and double mutants. All NS5A RAS in the GT1b replicon conferred VEL resistance < 3.3-fold. Most GT2a, GT3a, and GT4a single mutants showed low or no resistance to velpatasvir (VEL); however, the Y93H mutation in GT3a reduced VEL sensitivity by 723.5-fold. In summary, in a 3-day phase 1b monotherapy study, velpatasvir/VEL demonstrated broad genotypic activity and improved activity against pre-existing resistant variants. Although pre-treatment resistance-associated mutations (RAS) were associated with slightly reduced HCV RNA responses in GT1a and GT3 patients, current HCV treatment strategies based on the combination of direct-acting antiviral agents (DAAs) should be able to overcome any slight effects of pre-treatment RAS. Combination therapy with VEL and sofosbuvir (SOF) may provide an effective treatment option for patients infected with HCV GT1 to GT6. [2]

C49H54N8O8

883.00

882.406

C, 66.65; H, 6.16; N, 12.69; O, 14.49

1377049-84-7

67683363

White to light yellow solid powder.

1.3±0.1 g/cm3

1.643

6.78

193.1

4

10

13

65

1690

6

O(C([H])([H])[H])C([H])([H])[C@]1([H])C([H])([H])N(C([C@@]([H])(C2C([H])=C([H])C([H])=C([H])C=2[H])N([H])C(=O)OC([H])([H])[H])=O)[C@]([H])(C2=NC([H])=C(C3C([H])=C([H])C4=C(C([H])([H])OC5=C4C([H])=C4C([H])=C([H])C6=C(C4=C5[H])N=C([C@]4([H])C([H])([H])C([H])([H])[C@]([H])(C([H])([H])[H])N4C([C@]([H])(C([H])(C([H])([H])[H])C([H])([H])[H])N([H])C(=O)OC([H])([H])[H])=O)N6[H])C=3[H])N2[H])C1([H])[H]

FHCUMDQMBHQXKK-CDIODLITSA-N

InChI=1S/C49H54N8O8/c1-26(2)41(54-48(60)63-5)47(59)57-27(3)12-17-38(57)45-51-36-16-14-30-20-35-33-15-13-31(19-32(33)25-65-40(35)21-34(30)43(36)53-45)37-22-50-44(52-37)39-18-28(24-62-4)23-56(39)46(58)42(55-49(61)64-6)29-10-8-7-9-11-29/h7-11,13-16,19-22,26-28,38-39,41-42H,12,17-18,23-25H2,1-6H3,(H,50,52)(H,51,53)(H,54,60)(H,55,61)/t27-,28-,38-,39-,41-,42+/m0/s1

methyl ((R)-2-((2S,4S)-2-(5-(2-((2S,5S)-1-((methoxycarbonyl)-L-valyl)-5-methylpyrrolidin-2-yl)-1,11-dihydroisochromeno[4',3':6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl)-4-(methoxymethyl)pyrrolidin-1-yl)-2-oxo-1-phenylethyl)carbamate

GS5816; GS-5816; Velpatasvir; 1377049-84-7; GS5816; KCU0C7RS7Z; Velpatasvir [USAN:INN]; Velpatasvir [INN]; GS 5816; Velpatasvir

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : 100~146.66 mg/mL ( 113.25~166.09 mM )
Water : ~100 mg/mL
Ethanol : ~100 mg/mL

配方 1 中的溶解度: ≥ 2.5 mg/mL (2.83 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: 2.5 mg/mL (2.83 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (2.83 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: ≥ 2.5 mg/mL (2.83 mM)

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。