GT1a(EC50=34 pM);GT1b(EC50=4 pM)

体外活性：Ledipasvir（也称为 GS5885）是一种 HCV NS5A 聚合酶抑制剂，用于治疗丙型肝炎病毒感染。雷迪帕韦90毫克/索磷布韦400毫克的组合产品（商品名Harvoni）于2014年10月获得FDA批准。雷迪帕韦/索磷布韦组合是一种干扰HCV复制的直接作用抗病毒药物，可用于治疗基因型患者1a或1b，不含PEG-干扰素或利巴韦林。 Ledipasvir 在健康志愿者中的血浆半衰期延长至 37-45 小时，并且在基因型 1a HCV 感染患者中，每日一次口服 3 mg 剂量或更高剂量的单药治疗可产生超过 3 log 的病毒载量快速降低。它已被证明是安全有效的，与具有互补机制的直接作用抗病毒药物联合使用时，SVR12 率高达 100%。激酶测定：GT1a 复制子 EC50 = 31 pM 细胞测定：雷迪帕韦是 HCV NS5A 蛋白的特异性抑制剂，可抑制 HCV 亚基因组复制子系统中的 HCV 复制。 NS5A 复制复合物抑制剂是用于 HCV 治疗的新型抗病毒因子。通常，与针对 NS3 解旋酶和 NS5B RNA 聚合酶的传统 HCV 复制抑制剂相比，这些抑制剂具有高效率和低病毒耐药性。 NS5A 抑制剂应该跨 NS5A 二聚体界面结合，靠近 N 端结构域 1。这种结合被认为会直接或变构地扭曲二聚体关联，这可能会破坏 HCV RNA 复制中的 NS5A 功能。当使用 JFH1/3a-NS5A 杂合复制子评估对 NS5A 的敏感性时，另一种抑制剂 DCV 被证明比雷迪帕韦更有效。此外，NS5A-A30K和-Y93H变体表现出对ledpasvir的敏感性降低（EC50值分别为1770 nM和4300 nM）。

在临床试验中，观察到 ledpasvir 具有良好的耐受性，并且 HCV RNA 的中位最大减少范围为 2.3 log10 IU/ml 至 3.3 log10 IU/ml。 Emax 模型还显示，3 天后给予 30 mg ledpasvir 导致 HCV RNA 减少对基因型 1a 的最大反应大于 95%。最后，还观察到与 1a 相比，HCV RNA 在基因型 1b 中更持久。

竞争性蛋白结合试验[1]
将含有10%胎牛血清（CCM）的人血浆和细胞培养基以2μM的终浓度掺入受试化合物。将加标血浆（1 mL）和CCM（1 mL）放入组装好的透析细胞的相对侧，用半透膜隔开。透析细胞在37°C水浴中缓慢旋转达到平衡所需的时间。测量透析后血浆和CCM重量，并用LC/MS/MS测定血浆和CCM中的试验化合物浓度。

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代谢稳定性[1]
使用合并的肝微粒体组分（最终蛋白质浓度为0.5mg/mL）在3μM的最终试验化合物浓度下测定体外代谢稳定性。通过加入NADPH再生系统引发反应。在不同时间点将25μL的反应混合物等分转移到含有淬火溶液的板上。反应混合物中的试验化合物浓度用LC/MS/MS测定。肝脏固有清除率如Obach之前所述计算，预测清除率使用搅拌良好的肝脏模型计算，不受蛋白质限制。
还使用氚化测试化合物在冷冻保存的肝细胞中测定了代谢稳定性。孵育混合物含有1×106个肝细胞/mL和1μM氚化试验化合物（2.5μCi）。在37°C的温度下，在95%空气/5%二氧化碳（v/v）的潮湿环境中轻轻摇晃进行孵化。在0、1、3和6小时后取出50μL的等分试样，并将其加入100μL的淬火溶液中。在与HPLC系统耦合的流动闪烁无线电探测器上分析样品。代谢物根据放射性检测器的峰面积进行定量，无细胞对照样品用作参考。通过测量测试化合物的消失率，即形成的放射性标记代谢物和测试化合物的总峰面积的百分比，来确定肝细胞中的代谢稳定性。

Ledipasvir是HCV NS5A蛋白的特异性抑制剂，可抑制HCV亚基因组复制子系统中的HCV复制。 NS5A 复制复合物抑制剂是用于 HCV 治疗的新型抗病毒因子。通常，与针对 NS3 解旋酶和 NS5B RNA 聚合酶的传统 HCV 复制抑制剂相比，这些抑制剂具有高效率和低病毒耐药性。 NS5A 抑制剂应该跨 NS5A 二聚体界面结合，靠近 N 端结构域 1。这种结合被认为会直接或变构地扭曲二聚体关联，这可能会破坏 HCV RNA 复制中的 NS5A 功能。当使用 JFH1/3a-NS5A 杂合复制子评估对 NS5A 的敏感性时，另一种抑制剂 DCV 被证明比雷迪帕韦更有效。此外，NS5A-A30K和-Y93H变体表现出对ledpasvir的敏感性降低（EC50值分别为1770 nM和4300 nM）。

Absorption, Distribution and Excretion
Absorption
When given orally, ledipasvir reaches its maximum plasma concentration in about 4 to 4.5 hours with a maximum concentration (Cmax) of 323 ng/mL.

Route of Elimination
Following a single 90 mg oral dose of [14C]-ledipasvir, mean total recovery of the [14C]-radioactivity in feces and urine was approximately 87%, with most of the radioactive dose recovered from feces (approximately 86%). Unchanged ledipasvir excreted in feces accounted for a mean of 70% of the administered dose and the oxidative metabolite M19 accounted for 2.2% of the dose. These data indicate that biliary excretion of unchanged ledipasvir is a major route of elimination, with renal excretion being a minor pathway (approximately 1%).

Metabolism / Metabolites
In vitro, no detectable metabolism of ledipasvir was observed by human CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4. Evidence of slow oxidative metabolism via an unknown mechanism has been observed. Following a single dose of 90 mg [14C]-ledipasvir, systemic exposure was almost exclusively to the parent drug (>98%). Unchanged ledipasvir is the major species present in feces.

Biological Half-Life
The median terminal half-life of ledipasvir is 47 hours.

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
Ledipasvir has not been studied in nursing mothers being treated for hepatitis C infection. Because it is 99.8% bound to maternal plasma proteins, amounts in breastmilk are likely to be very low. If ledipasvir alone or in combination with sofosbuvir (Harvoni) is required by the mother, it is not a reason to discontinue breastfeeding. Some sources recommend against breastfeeding when ledipasvir is used with ribavirin.

Hepatitis C is not transmitted through breastmilk and breastmilk has been shown to inactivate hepatitis C virus (HCV). However, the Centers for Disease Control recommends that mothers with HCV infection should consider abstaining from breastfeeding if their nipples are cracked or bleeding. It is not clear if this warning would apply to mothers who are being treated for hepatitis C.

Infants born to mothers with HCV infection should be tested for HCV infection; because maternal antibody is present for the first 18 months of life and before the infant mounts an immunologic response, nucleic acid testing is recommended. ◉ 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.
Drugs and Lactation Database (LactMed)

Protein Binding
Ledipasvir is >99.8% bound to human plasma proteins.

[1]. Discovery of ledipasvir (GS-5885): a potent, once-daily oral NS5A inhibitor for the treatment of hepatitis C virus infection. J Med Chem. 2014 Mar 13;57(5):2033-46

[2]. Natural prevalence of NS5A polymorphisms in subjects infected with hepatitis C virus genotype 3 and their effects on the antiviral activity of NS5A inhibitors. J Clin Virol. 2013 May;57(1):13-8.

A new class of highly potent NS5A inhibitors with an unsymmetric benzimidazole-difluorofluorene-imidazole core and distal [2.2.1]azabicyclic ring system was discovered. Optimization of antiviral potency and pharmacokinetics led to the identification of 39 (ledipasvir, GS-5885). Compound 39 (GT1a replicon EC50 = 31 pM) has an extended plasma half-life of 37-45 h in healthy volunteers and produces a rapid >3 log viral load reduction in monotherapy at oral doses of 3 mg or greater with once-daily dosing in genotype 1a HCV-infected patients. 39 has been shown to be safe and efficacious, with SVR12 rates up to 100% when used in combination with direct-acting antivirals having complementary mechanisms.[1]

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Background: Hepatitis C virus (HCV) NS5A replication complex inhibitors (RCIs) have been shown to exhibit picomolar antiviral activity against genotype 1 (GT1) in vitro. This has translated into rapid and robust declines in HCV RNA in GT1 patients. Less is known about the susceptibility of other genotypes such as GT3 to inhibition by NS5A RCIs. Objectives: To detect and phenotype naturally occurring HCVGT3 NS5A polymorphisms against two NS5A RCIs (daclatasvir [DCV] and GS-5885) currently in clinical development. Study design: The NS5A region from 96 HCV GT3 treatment-naive patients spanning North America, Europe and Australia was determined. Results: Phylogenetic analysis revealed a broad distribution with no significant geographic clustering. GT1 DCV resistance-associated variants (RAVs) were observed in GT3 subjects; variants (and their frequencies) included 28M/V (1%), 30A/K/S/T/V (10%), 31L/M (1%), E92A (1%) and Y93H (8.3%). A consensus sequence was used to generate a JFH1/3a-NS5A hybrid replicon and employed to assess susceptibility to NS5A RCIs. Against JFH1/3a-NS5A, DCV was more potent (EC(50) = 0.52 nM) than GS-5885 (EC(50) = 141 nM). DCV sensitivity was increased against JFH1/3a-NS5A-M28V (EC50 = 0.006 nM), A30V (EC(50) = 0.012 nM), and E92A (EC(50) = 0.004 nM) while the NS5A-A30K and -Y93H variants exhibited reduced sensitivity to DCV (EC50 values of 23 nM and 1120 nM, respectively) and to GS-5885 (EC50 values of 1770 nM and 4300 nM, respectively). Conclusions: Substitutions conferring resistance to NS5A RCIs pre-existed in treatment-naive patients infected with HCV GT3. The effectiveness of these NS5A RCIs to exert efficacy in the clinic may depend on which inhibitor is used in combination with other antivirals.[2]

C53H60F2N8O12

1039.09

1038.43

1502654-87-6

Ledipasvir;1256388-51-8;Ledipasvir (acetone);1441674-54-9;Ledipasvir-d6;2050041-12-6;Ledipasvir hydrochloride;2128695-48-5;Ledipasvir (diacetone);1502655-48-2

78357794

Off-white to yellow solid powder

7.142

289.7

CC(C)[C@@H](C(=O)N1CC2(CC2)C[C@H]1C3=NC=C(N3)C4=CC5=C(C=C4)C6=C(C5(F)F)C=C(C=C6)C7=CC8=C(C=C7)N=C(N8)[C@@H]9[C@H]1CC[C@H](C1)N9C(=O)[C@H](C(C)C)NC(=O)OC)NC(=O)OC.[C@H]([C@@H](C(=O)O)O)(C(=O)O)O

ZQVLPYMRXLPMDX-KEAIDYLOSA-N

InChI=1S/C49H54F2N8O6.C4H6O6/c1-24(2)39(56-46(62)64-5)44(60)58-23-48(15-16-48)21-38(58)42-52-22-37(55-42)28-9-13-32-31-12-8-26(18-33(31)49(50,51)34(32)19-28)27-10-14-35-36(20-27)54-43(53-35)41-29-7-11-30(17-29)59(41)45(61)40(25(3)4)57-47(63)65-6;5-1(3(7)8)2(6)4(9)10/h8-10,12-14,18-20,22,24-25,29-30,38-41H,7,11,15-17,21,23H2,1-6H3,(H,52,55)(H,53,54)(H,56,62)(H,57,63);1-2,5-6H,(H,7,8)(H,9,10)/t29-,30+,38-,39-,40-,41-;1-,2-/m00/s1

Methyl N-[(2S)-1-[(6S)-6-[5-[9,9-Difluoro-7-[2-[(1S,2S,4R)-3-[(2S)-2-(methoxycarbonylamino)-3-methylbutanoyl]-3-azabicyclo[2.2.1]heptan-2-yl]-3H-benzimidazol-5-yl]fluoren-2-yl]-1H-imidazol-2-yl]-5-azaspiro[2.4]heptan-5-yl]-3-methyl-1-oxobutan-2-yl]carbamate tartrate

GS-5885 tartrate, GS5885 tartrate; GS 5885; Ledipasvir D-tartrate; Ledipasvir (D-tartrate); 1502654-87-6; RT680T6HCQ; 1499193-68-8; UNII-RT680T6HCQ; (2S,3S)-2,3-dihydroxybutanedioic acid;methyl N-[(2S)-1-[(6S)-6-[5-[9,9-difluoro-7-[2-[(1R,3S,4S)-2-[(2S)-2-(methoxycarbonylamino)-3-methylbutanoyl]-2-azabicyclo[2.2.1]heptan-3-yl]-3H-benzimidazol-5-yl]fluoren-2-yl]-1H-imidazol-2-yl]-5-azaspiro[2.4]heptan-5-yl]-3-methyl-1-oxobutan-2-yl]carbamate; trade name: Harvoni;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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

DMSO : ~25 mg/mL (~24.06 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (2.41 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 (2.41 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 储备液添加到 900 μL 玉米油中并混合均匀。

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配方 3 中的溶解度: 10% DMSO+90% Corn Oil: ≥ 2.5 mg/mL (2.41 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。