genotype 1 HCV NS3-4A protease (Ki = 7 nM)

体外活性：特拉匹韦抑制丙型肝炎病毒 NS3-4A 丝氨酸蛋白酶，导致病毒多蛋白加工受阻，随后降低 Con1（基因型 1b）亚基因组 HCV 复制子细胞中的病毒 RNA 复制、总 HCV RNA 水平和蛋白质水平以时间和剂量依赖性方式。 Telaprevir 对 HCV RNA 复制的抑制作用表现出显着的时间依赖性增强，孵育 24、48、72 和 120 小时的 IC50 值分别为 0.574 μM、0.488 μM、0.210 μM 和 0.139 μM。根据使用 48 小时孵育的三个独立实验，Telaprevir 显示的平均 IC50 为 0.354 μM，平均 IC90 为 0.830 μM。孵育48小时后，特拉匹韦对HCV复制子细胞、亲本Huh-7和HepG2细胞没有明显的细胞毒性。 Telaprevir (17.5 μM) 孵育 13 天后，可完全消除复制子细胞中的 HCV RNA，且撤除 Telaprevir 后不会出现反弹。与单独使用每种药物治疗相比，特拉匹韦与 IFN-α 联合使用时，在减少 HCV RNA 复制和抑制耐药突变方面表现出附加至中度的协同作用，且细胞毒性没有显着增加。激酶测定：含有自我复制、亚基因组 HCV 复制子（其序列与 I377neo/NS3-3/wt 复制子相同）的稳定 Huh-7 细胞用于抗 HCV 测定。将复制子细胞与在含有 2% FBS 和 0.5% 二甲基亚砜 (DMSO) 的 DMEM 中连续稀释的 Telaprevir 一起在 37°C 下孵育指定的时间。使用 RNeasy-96 试剂盒提取总细胞 RNA，并使用定量 RTPCR (QRT-PCR) 测定法测定 HCV RNA 的拷贝数，以评估 50% 抑制浓度 (IC50)。细胞测定：将细胞（Huh-7、HepG2 和外周血单核细胞 (PBMC)）与不同浓度的 Telaprevir 一起孵育 48 小时。细胞活力通过使用基于四唑 (MTS) 的细胞活力测定来测定。

在小鼠模型中，口服 Telaprevir 在剂量为 10 和 25 mg/kg 时，可将 HCV 蛋白酶依赖性裂解和随后 SEAP 从肝脏分泌到血液中的量分别减少至 18.7% 和 18.4%。给予特拉匹韦 200 mg/kg 1 周，导致基因型 1b HCV 感染的人肝细胞嵌合小鼠的 HCV RNA 减少 1.9 个对数，当与 MK-0608 (50 mg/kg) 联合治疗 4 周时，病毒从小鼠身上被消灭。

含有自我复制、亚基因组 HCV 复制子（其序列与 I377neo/NS3-3'/wt 复制子相同）的稳定 Huh-7 细胞用于抗 HCV 测定。将特拉匹韦在含有 2% FBS 和 0.5% 二甲基亚砜 (DMSO) 的 DMEM 中连续稀释，与复制细胞在 37 °C 下孵育指定的时间。使用 RNeasy-96 试剂盒提取细胞总 RNA，并使用定量实时聚合酶链反应 (QRT-PCR) 测定法测定 HCV RNA 的拷贝数，以评估 50% 抑制浓度 (IC50) 。

在 HCV 复制子细胞中评估 Telaprevir (VX-950) 或 IFN-α 涉及确定其 IC50、IC90 和 CC50。综上所述，96孔板每孔铺有1×10⁴复制子细胞。使用在 DMEM 加 2% FBS 和 0.5% DMSO 中连续稀释的抗病毒剂，复制子细胞在第二天在 37°C 下孵育指定的时间。使用 RNeasy-96 试剂盒提取总细胞 RNA，并使用定量 RT-PCR (QRT-PCR) 测定法计算 HCV RNA 的拷贝数。每个数据点代表细胞培养物中五次重复的平均值。在相同的实验条件下，使用基于四唑（MTS）的细胞活力测定来测量 Telaprevir 的细胞毒性。每孔 100 万个亲本 Huh-7 细胞或 400 万个 HepG2 细胞用于使用人肝细胞系的细胞毒性测定。为了评估 Telaprevir 对静息外周血单克隆细胞的细胞毒性，每孔 1×10⁵ 细胞，在 RPMI-1640 培养基（无血清）中与 Telaprevir 一起培养 48 小时，之后 MTS基于的测定用于确定细胞活力。预涂有抗人 CD3 抗体的 96 孔板，每孔在 RPMI-1640 培养基中填充 1×10⁵ 细胞，以测试 VX-950 对增殖的 PBMC 的细胞毒性。将细胞与 Telaprevir 和抗人 CD28 抗体一起在 37°C 下培养 72 小时。 [³H]胸苷更新用于测量第 48 小时和第 72 小时之间的细胞生长[1]。

Mice: Recombinant adenovirus Ad-WT-HCVpro-SEAP, with 10⁹ IFU per mouse, is injected via the tail vein into five groups of six-week-old SCID mice (six animals per group). Two oral doses of Telaprevir (VX-950) at a dose of 10, 25, 75, 150, or 300 mg/kg are administered to each group of mice. First dose of Telaprevir is administered two hours prior to adenovirus injection; second dose is administered ten hours following injection. A second set of ten mice is given the vehicle on its own. Serum samples are taken 24 hours after injection, and the SEAP activity in each group administered with Telaprevir is contrasted with the vehicle group's. Rat and Canine Rats and dogs are used to assess the oral and intravenous pharmacokinetics of telaprevir (VX-950). One intravenous bolus dose of 0.95 mg/kg Telaprevir is given intravenously to three male Sprague-Dawley rats weighing 250–300 g. Heparinized tubes are used to collect serial blood samples prior to dosage administration and at intervals of 0.083, 0.167, 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, and 8 hours following the dose. Telaprevir in 10% ethanol, 40% polyethylene glycol 400, and 50% D5W is given intravenously as a bolus dose to three male beagle dogs (8–12 kg). Heparinized tubes are used to collect serial blood samples prior to dosage administration as well as at 0.083, 0.167, 0.25, 0.5, 1, 1.5, 2, 4, 6, 8, 12, and 24 hours later. Telaprevir is formulated in polyvinylpyrrolidone (PVP) K-30 plus 2% sodium lauryl sulfate and dosed as an oral gavage for oral studies in rats and dogs. Oral dosages of 40 mg/kg VX-950 are given to three male Sprague-Dawley rats (250–300 g) and 9.6 mg/kg VX-950 are given to four male Beagle dogs (10.9–12.0 kg). Blood samples are obtained before dosage administration and at 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, and 24 hours following dose administration in both oral studies. Plasma samples are obtained by centrifugation and kept at -70°C until analysis in both intravenous and oral studies. Samples from the oral studies are analyzed using an achiral LC/MS/MS method, while samples from the intravenous studies are analyzed using a chiral liquid chromatography followed by tandem mass spectrometry (LC/MS/MS) method.

Absorption, Distribution and Excretion
Telaprevir reaches peak plasma concentration 4-5hours after administration. Absolute bioavailability has not been determined. When taken with a normal fat meal (21g of fat), exposure increases by 235% compared to fasting conditions. With low (3.6g of fat) and high fat (56g of fat) meals, exposure increased 117% and 330% respectively.
Telaprevir is mainly eliminated in the feces (82%) with a smaller amount eliminated via expiration (9%) and very little in the urine (1%). 31.9% and 18.8% of drug in the feces was present as the parent compound and R-diastereomer of the parent compound respectively.
The estimated apparent volume of distribution for Telapravir is 252 litres with an inter-individual variability of 72%.
Telaprevir has an estimated aparent total body clearance of 32.4 liters per hour with an interindividual variability of 27.2%.
The pharmacokinetic properties of telaprevir have been evaluated in healthy adult subjects and in subjects with chronic hepatitis C. Following multiple doses of telaprevir (750 mg every 8 hr) in combination with peginterferon alfa and ribavirin in treatment-naive subjects with genotype 1 chronic hepatitis C, mean (SD) Cmax was 3510 (1280) ng/mL, Cmin was 2030 (930) ng/mL, and AUC8h was 22,300 (8650) ng.hr/mL.
Telaprevir is orally available, most likely absorbed in the small intestine, with no evidence for absorption in the colon. Maximum plasma concentrations after a single dose of telaprevir are generally achieved after 4 to 5 hours.
Telaprevir is a substrate for and inhibitor of P-glycoprotein transport.
The systemic exposure (AUC) to telaprevir was increased by 237% when telaprevir was administered with a standard fat meal (containing 533 kcal and 21 g fat) compared to when telaprevir was administered under fasting conditions. In addition, the type of meal significantly affects exposure to telaprevir. Relative to fasting, when telaprevir was administered with a low-fat meal (249 kcal, 3.6 g fat) and a high-fat meal (928 kcal, 56 g fat), the systemic exposure (AUC) to telaprevir was increased by approximately 117% and 330%, respectively.
For more Absorption, Distribution and Excretion (Complete) data for Telaprevir (13 total), please visit the HSDB record page.

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Metabolism / Metabolites
Telaprevir is extensively metabolized via hydrolysis, oxidation, and reduction. The major metabolites of Telaprevir are pyrazinoic acid, a metabolite that underwent reduction at the α-ketoamide bond, and the R-diastereomer of telaprevir which is 30-fold less active than the parent compound were found to be the predominant metabolites. The primary enzyme involved in the metabolism of Telaprevir is CYP3A4. Some metabolism is performed by aldo-keto reductases and other reductases.
Telaprevir is extensively metabolized in the liver, involving hydrolysis, oxidation, and reduction. Multiple metabolites were detected in feces, plasma, and urine. After repeated oral administration, the R-diastereomer of telaprevir (30-fold less active), pyrazinoic acid, and a metabolite that underwent reduction at the alpha-ketoamide bond of telaprevir (not active) were found to be the predominant metabolites of telaprevir.

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Biological Half-Life
Telaprevir has a half-life of elimination of 4.0-4.7 hours after a single dose and an effective half life of 9-11 hours at steady state.
The mean elimination half-life after single-dose oral administration of telaprevir 750 mg typically ranged from about 4.0 to 4.7 hours. At steady state, the effective half-life is about 9 to 11 hours.

Hepatotoxicity
In large randomized controlled trials, triple therapy with telaprevir, peginterferon and ribavirin was associated with a high rate of adverse events that often required dose adjustments and led to early discontinuation in 5% to 20% of patients. However, serum ALT elevations and clinically apparent liver injury were not generally mentioned as adverse events of therapy. Telaprevir, however, was associated with a high rate of rash, which was sometimes associated with features of hypersensitivity, including rare instances of DRESS and Stevens Johnson syndrome. These severe cutaneous reactions are often accompanied by laboratory evidence of hepatic injury (ALT and alkaline phosphatase elevations). In reported cases, however, the rash and other features of hypersensitivity typically overshadowed the hepatic injury and none were reported to be associated with jaundice.
Another rare but severe hepatic complications of telaprevir therapy occurs in patients with advanced fibrosis or cirrhosis, among whom de novo, seemingly spontaneous hepatic decompensation occurred in a proportion of treated subjects. Decompensation was particularly common in patients with advanced fibrosis or cirrhosis with a previous history of decompensation. The cause of the decompensation was not clear and the separate role of telaprevir in contrast to peginterferon and ribavirin could not be defined. Nevertheless, in postmarketing studies of triple therapy of chronic hepatitis C with cirrhosis, decompensation was reported in 2% to 8% of patients, and deaths from hepatic failure in 1% to 3%.
Likelihood score for the combination of telaprevir, peginterferon and ribavirin: B (likely cause of liver injury and hepatic decompensation in patients with preexisting cirrhosis or advanced fibrosis).

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Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
Telaprevir is no longer marketed in the United States and has not been studied in nursing mothers. Because it must be used with ribavirin and peginterferon alfa, it is not considered a good choice during breastfeeding. When it was marketed, the manufacturer recommended that mothers taking telaprevir not breastfeed their infants.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

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Protein Binding
Telapravir is 59-76% bound to human plasma proteins following a single dose. It binds to both human serum albumin and α1-acid glycoprotein.

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Interactions
Telaprevir is a strong inhibitor of CYP3A. Telaprevir is contraindicated when combined with drugs that are highly dependent on CYP3A for clearance and for which elevated plasma concentrations are associated with serious and/or life-threatening events (narrow therapeutic index). Telaprevir is contraindicated when combined with drugs that strongly induce CYP3A and thus may lead to lower exposure and loss of efficacy of telaprevir.
Potential pharmacokinetic interaction with drugs that are inducers or inhibitors of P-glycoprotein, with possible alteration in telaprevir concentrations.
Potential pharmacokinetic interaction with alfuzosin (increased alfuzosin concentrations). Concomitant use of telaprevir and alfuzosin is contraindicated because increased alfuzosin concentrations may result in hypotension or cardiac arrhythmia.
Potential pharmacokinetic interaction with antiarrhythmic agents (amiodarone, bepridil (no longer commercially available in US), flecainide, systemic lidocaine, propafenone, quinidine) may result in increased concentrations of the antiarrhythmic agent; potential for serious and/or life-threatening adverse effects. If telaprevir and antiarrhythmic agents are used concomitantly, use caution and clinical monitoring.
For more Interactions (Complete) data for Telaprevir (54 total), please visit the HSDB record page.

[1]. Antimicrob Agents Chemother . 2006 May;50(5):1813-22.

[2]. Antimicrob Agents Chemother . 2006 Mar;50(3):899-909.

[3]. J Hepatol . 2011 May;54(5):872-8.

Therapeutic Uses
Oligopeptides
INCIVEK (telaprevir), in combination with peginterferon alfa and ribavirin, is indicated for the treatment of genotype 1 chronic hepatitis C in adult patients with compensated liver disease, including cirrhosis, who are treatment-naive or who have previously been treated with interferon-based treatment, including prior null responders, partial responders, and relapsers. /Included in US product label/

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Drug Warnings
Fatal and non-fatal serious skin reactions, including Stevens Johnson Syndrome (SJS), Drug Reaction with Eosinophilia and Systemic Symptoms (DRESS), and Toxic Epidermal Necrolysis (TEN), have been reported in patients treated with INCIVEK combination treatment. Fatal cases have been reported in patients with progressive rash and systemic symptoms who continued to receive INCIVEK combination treatment after a serious skin reaction was identified. For serious skin reactions, including rash with systemic symptoms or a progressive severe rash, INCIVEK, peginterferon alfa, and ribavirin must be discontinued immediately. Discontinuing other medications known to be associated with serious skin reactions should be considered. Patients should be promptly referred for urgent medical care.
Rash developed in 56% of patients receiving telaprevir during controlled clinical trials. Severe rash (e.g., generalized rash or rash with vesicles or bullae or ulcerations other than SJS) was reported in 4% of patients receiving telaprevir in conjunction with peginterferon alfa and ribavirin compared with less than 1% of patients receiving peginterferon alfa and ribavirin without telaprevir. Rash frequently was observed during the first 4 weeks of telaprevir treatment, but can occur at any time. Rash generally improves when telaprevir therapy is completed or discontinued; complete resolution may take weeks.
If a serious skin reaction occurs, telaprevir, peginterferon alfa, and ribavirin should be immediately discontinued and the patient promptly referred for urgent medical care. Patients with mild to moderate rash should be monitored for progression of rash or development of systemic symptoms. If rash progresses and becomes severe or if systemic symptoms develop, telaprevir should be discontinued; peginterferon alfa and ribavirin may be continued.
Telaprevir dosage should not be reduced and telaprevir should not be restarted if it was discontinued because of rash. If improvement is not observed within 7 days of discontinuing telaprevir, sequential or simultaneous interruption or discontinuance of peginterferon alfa and/or ribavirin should be considered. If medically indicated, earlier interruption or discontinuance of peginterferon alfa and ribavirin should be considered.
For more Drug Warnings (Complete) data for Telaprevir (18 total), please visit the HSDB record page.

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Pharmacodynamics
Telaprevir is classified as a direct-acting antiviral (DAA) and prevents viral replication in HCV genotype 1.

C36H53N7O6

679.85

679.405

C, 63.60; H, 7.86; N, 14.42; O, 14.12

402957-28-2

Telaprevir-d4

3010818

White to off-white solid powder

1.3±0.1 g/cm3

1.584

3.93

179.56

4

8

14

49

1240

6

O=C(N([C@@H]1C(N[C@H](C(C(NC2CC2)=O)=O)CCC)=O)C[C@@]3(CCC[C@@]31[H])[H])[C@@H](NC([C@@H](NC(C4=NC=CN=C4)=O)C5CCCCC5)=O)C(C)(C)C

BBAWEDCPNXPBQM-GDEBMMAJSA-N

InChI=1S/C36H53N7O6/c1-5-10-25(29(44)34(48)39-23-15-16-23)40-33(47)28-24-14-9-13-22(24)20-43(28)35(49)30(36(2,3)4)42-32(46)27(21-11-7-6-8-12-21)41-31(45)26-19-37-17-18-38-26/h17-19,21-25,27-28,30H,5-16,20H2,1-4H3,(H,39,48)(H,40,47)(H,41,45)(H,42,46)/t22-,24-,25-,27-,28-,30+/m0/s1

(3S,3aS,6aR)-2-[(2S)-2-[[(2S)-2-cyclohexyl-2-(pyrazine-2-carbonylamino)acetyl]amino]-3,3-dimethylbutanoyl]-N-[(3S)-1-(cyclopropylamino)-1,2-dioxohexan-3-yl]-3,3a,4,5,6,6a-hexahydro-1H-cyclopenta[c]pyrrole-3-carboxamide

VX-950; LY-570310; MP-424; VX950; LY570310; MP424; VX 950; LY 570310; MP 424; trade names: Incivek; Incivo

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)

配方 1 中的溶解度: ≥ 2.5 mg/mL (3.68 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900 μL 玉米油中，混合均匀。

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配方 2 中的溶解度: 30% PEG400+0.5% Tween80+5% propylene glycol: 30 mg/mL

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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。