| 规格 | 价格 | 库存 | 数量 |
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| 靶点 |
SARS-CoV-2 RNA-dependent RNA polymerase (RdRp), with inhibitory activity mediated through its triphosphate form (GS-443902). The inhibition mechanism involves delayed chain termination during viral RNA synthesis. The half-maximal inhibitory concentration (IC50) for SARS-CoV-2 RdRp is 0.0038 μM (measured via enzymatic assay with purified polymerase complex) [3]
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| 体外研究 (In Vitro) |
- Vero E6细胞抗病毒活性:GS-704277(在细胞内水解为GS-441524)对SARS-CoV-2的EC50为 1.65 μM,通过病毒RNA定量和细胞病变效应抑制实验测定。氯喹(EC50 = 1.13 μM)作为阳性对照 [2]
- 作用机制:GS-704277在细胞内经磷酸化转化为活性三磷酸形式GS-443902,与ATP竞争结合RdRp,导致病毒RNA合成提前终止。该机制经Huh-7细胞RNA延伸实验证实 [3] .进行了标准检测,以测量这些化合物对2019-nCoVs的细胞毒性、病毒产量和感染率的影响。首先,通过CCK8试验测定候选化合物在Vero E6细胞(ATCC-1586)中的细胞毒性。然后,在不同浓度的试验药物存在下,以0.05的感染复数(MOI)用nCoV-2019BetaCoV/WWuhan/WIV04/20192感染Vero E6细胞。对照组使用DMSO。通过定量实时RT-PCR(qRT-PCR)定量细胞上清液中的病毒拷贝数来评估疗效,并在感染后48小时通过免疫荧光显微镜观察病毒核蛋白(NP)的表达来证实(感染时细胞病变效应不明显)。在七种受试药物中,需要高浓度的三种核苷类似物,包括利巴韦林(半数最大有效浓度(EC50)=109.50μM,半数细胞毒性浓度(CC50)>400μM,选择性指数(SI)>3.65)、喷昔洛韦(EC50=95.96μM,CC50>400μM,SI>4.17)和法匹拉韦(EC50=61.88μM,CC 50>400µM,SI>6.46),以减少病毒感染(图1a和补充信息,图S1)。然而,favipiravir已被证明对保护小鼠免受埃博拉病毒攻击100%有效,尽管其在Vero E6细胞中的EC50值高达67μM,4这表明建议进行进一步的体内研究来评估这种抗病毒核苷。Nafamostat是MERS-CoV的强效抑制剂,可防止膜融合,对2019-nCoV感染具有抑制作用(EC50=22.50μM,CC50>100μM,SI>4.44)。硝唑尼特是一种商业抗原生动物药物,对包括人类和动物冠状病毒在内的多种病毒具有抗病毒潜力,在低微摩尔浓度下抑制2019-nCoV(EC50=2.12μM;CC50>35.53μM;SI>16.76)。建议对该药物进行进一步的体内评估,以对抗2019-nCoV感染。值得注意的是,两种化合物瑞德西韦(EC50=0.77μM;CC50>100μM;SI>129.87)和氯喹(EC50=1.13μM;CC50>100μM,SI>88.50)在低微摩尔浓度下有效阻断病毒感染,并显示出高SI(图1a,b)[1]。 |
| 酶活实验 |
- RdRp抑制实验:纯化的SARS-CoV-2 RdRp复合物(nsp7/nsp8/nsp12)与模板引物RNA、NTP(含[α-32P]ATP)及不同浓度GS-443902(GS-704277活性形式)孵育。定时终止反应后,通过凝胶电泳分离RNA产物,经光密度定量分析全长RNA条带,计算IC50 [3]
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| 细胞实验 |
- 抗病毒细胞病变效应(CPE)实验:Vero E6细胞感染SARS-CoV-2(MOI = 0.002)后,用GS-704277梯度浓度处理48小时。采用细胞计数试剂盒检测细胞活性,同时取上清液提取病毒RNA进行RT-qPCR定量。EC50通过CPE抑制率和病毒RNA减少的剂量效应曲线计算 [2]
- 代谢活化实验:Huh-7细胞与GS-704277孵育24小时后,通过LC-MS/MS定量细胞内代谢物(GS-441524、GS-704277和GS-443902),证实其快速水解为GS-441524并进一步磷酸化为GS-443902 [3] |
| 药代性质 (ADME/PK) |
Metabolic pathway: GS-704277 is an alanine prodrug of GS-441524. In vivo, it is rapidly hydrolyzed by cathepsin A (CatA) and carboxylesterase 1 (CES1) to release the parent nucleoside analog GS-441524, which is then phosphorylated intracellularly to the active triphosphate GS-443902. This conversion has been validated in human hepatocytes and primary monocytes [1] - Systemic exposure: In rhesus monkeys intravenously injected with remdesivir (the parent drug), the plasma half-life (t1/2) of the metabolite GS-441524 (derived from GS-704277) was approximately 1.2 hours, and the volume of distribution (Vd) was approximately 30 L/kg, indicating its extensive tissue distribution [1]
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| 毒性/毒理 (Toxicokinetics/TK) |
In vitro cytotoxicity: At concentrations up to 100 μM, GS-704277 did not show significant cytotoxicity in Vero E6 cells (CC50 > 100 μM) [2] - Elevated liver enzymes: In a rhesus monkey model infected with MERS-CoV, remdesivir (producing GS-704277 metabolites) induced a transient increase in alanine aminotransferase (ALT) at high doses (10 mg/kg/day), but no histopathological liver damage was observed [1]
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| 参考文献 | |
| 其他信息 |
GS-704277 is an intermediate metabolite of remdesivir (GS-5734), generated by the hydrolysis of the parent prodrug. It is further metabolized to the nucleoside analog GS-441524, which is phosphorylated into the pharmacologically active triphosphate GS-443902 after entering the cell [1]. The efficacy of this compound depends on the activity of the viral corrected exonuclease (ExoN); viruses lacking ExoN (such as mouse hepatitis virus) are 10 times more sensitive to GS-443902 than coronaviruses with ExoN function [3]. The global pandemic of SARS-CoV-2 (the pathogen of COVID-19) has prompted the biomedical community to take action to explore and develop antiviral interventions. One potential treatment currently being evaluated in multiple clinical trials is remdesivir, whose development has been long and tortuous. Remdesivir is a nucleotide analog prodrug that interferes with viral replication and was initially used in clinical trials in 2014 to contain the Ebola outbreak. Subsequently, numerous virology laboratory evaluations confirmed remdesivir's ability to inhibit the replication of coronaviruses, including SARS-CoV-2. This article summarizes the discovery of remdesivir, its mechanism of action, and ongoing clinical efficacy studies.
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| 分子式 |
C15H19N6O8P
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|---|---|
| 分子量 |
442.320523500443
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| 精确质量 |
442.1
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| 元素分析 |
C, 40.73; H, 4.33; N, 19.00; O, 28.94; P, 7.00
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| CAS号 |
1911579-04-8
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| 相关CAS号 |
1355149-45-9 [GS443902 (GS-441524 triphosphate)]; 1809249-37-3 (Remdesivir); 1191237-69-0 (GS-441524, an active metabolite of Remdesivir); 1191237-80-5 (Remdesivir O-desphosphate acetonide impurity); 1911578-74-9 (Remdesivir nucleoside monophosphate); 1911579-04-8 (GS-704277)
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| PubChem CID |
121313150
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| 外观&性状 |
Typically exists as White to light yellow solids at room temperature
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| LogP |
-2.8
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| tPSA |
226Ų
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| 氢键供体(HBD)数目 |
6
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| 氢键受体(HBA)数目 |
13
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| 可旋转键数目(RBC) |
7
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| 重原子数目 |
30
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| 分子复杂度/Complexity |
765
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| 定义原子立体中心数目 |
5
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| SMILES |
C[C@@H](C(=O)O)NP(=O)(O)OC[C@@H]1[C@H]([C@H]([C@](O1)(C#N)C2=CC=C3N2N=CN=C3N)O)O
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| InChi Key |
IYHPTSNEWCZBDF-NIFWRESRSA-N
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| InChi Code |
InChI=1S/C15H19N6O8P/c1-7(14(24)25)20-30(26,27)28-4-9-11(22)12(23)15(5-16,29-9)10-3-2-8-13(17)18-6-19-21(8)10/h2-3,6-7,9,11-12,22-23H,4H2,1H3,(H,24,25)(H2,17,18,19)(H2,20,26,27)/t7-,9+,11+,12+,15-/m0/s1
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| 化学名 |
((((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)-L-alanine
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| 别名 |
GS-704277; GS704277; GS-704277; N-phosphono-L-alanine, P-->6-ester with 2-C-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-2,5-anhydro-D-altrononitrile; (2S)-2-[[[(2R,3S,4R,5R)-5-(4-Aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]amino]propanoic acid; Dephenoxy Hydroxy Remdesivir; Remdesivir Impurity 39; SCHEMBL17722667; MFCD32701941; GS 704277
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| HS Tariff Code |
2934.99.9001
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| 存储方式 |
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(例如氮气保护),避免吸湿/受潮。 |
| 运输条件 |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| 溶解度 (体外实验) |
DMSO : ~100 mg/mL (~226.08 mM)
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| 溶解度 (体内实验) |
注意: 如下所列的是一些常用的体内动物实验溶解配方,主要用于溶解难溶或不溶于水的产品(水溶度<1 mg/mL)。 建议您先取少量样品进行尝试,如该配方可行,再根据实验需求增加样品量。
注射用配方
注射用配方1: DMSO : Tween 80: Saline = 10 : 5 : 85 (如: 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)(IP/IV/IM/SC等) *生理盐水/Saline的制备:将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中,得到澄清溶液。 注射用配方 2: DMSO : PEG300 :Tween 80 : Saline = 10 : 40 : 5 : 45 (如: 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline) 注射用配方 3: DMSO : Corn oil = 10 : 90 (如: 100 μL DMSO → 900 μL Corn oil) 示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液,加到 900 μL Corn oil/玉米油中, 混合均匀。 View More
注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如:100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)] 口服配方
口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠) 口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素) 示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中,得到0.5%CMC-Na澄清溶液;然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中,得到悬浮液。 View More
口服配方 3: 溶解于 PEG400 (聚乙二醇400) 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。 |
| 制备储备液 | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2608 mL | 11.3040 mL | 22.6081 mL | |
| 5 mM | 0.4522 mL | 2.2608 mL | 4.5216 mL | |
| 10 mM | 0.2261 mL | 1.1304 mL | 2.2608 mL |
1、根据实验需要选择合适的溶剂配制储备液 (母液):对于大多数产品,InvivoChem推荐用DMSO配置母液 (比如:5、10、20mM或者10、20、50 mg/mL浓度),个别水溶性高的产品可直接溶于水。产品在DMSO 、水或其他溶剂中的具体溶解度详见上”溶解度 (体外)”部分;
2、如果您找不到您想要的溶解度信息,或者很难将产品溶解在溶液中,请联系我们;
3、建议使用下列计算器进行相关计算(摩尔浓度计算器、稀释计算器、分子量计算器、重组计算器等);
4、母液配好之后,将其分装到常规用量,并储存在-20°C或-80°C,尽量减少反复冻融循环。
计算结果:
工作液浓度: mg/mL;
DMSO母液配制方法: mg 药物溶于 μL DMSO溶液(母液浓度 mg/mL)。如该浓度超过该批次药物DMSO溶解度,请首先与我们联系。
体内配方配制方法:取 μL DMSO母液,加入 μL PEG300,混匀澄清后加入μL Tween 80,混匀澄清后加入 μL ddH2O,混匀澄清。
(1) 请确保溶液澄清之后,再加入下一种溶剂 (助溶剂) 。可利用涡旋、超声或水浴加热等方法助溶;
(2) 一定要按顺序加入溶剂 (助溶剂) 。