HIV-1/2 nucleotide reverse transcriptase

富马酸替诺福韦艾拉酚胺（GS-7340 富马酸盐）在所有细胞类型中均表现出相当的抗病毒活性，对于 MT-4 和 MT-2 细胞，CC50 范围分别为 4.7 至 42 μM。富马酸盐的浓度范围为 5 至 7 nM。一组 HIV-1 和 HIV-2 分离株（包括 HIV-1 M A 至 G 亚型以及 N 和 O 组分离株）用于评估 TAF 的抗病毒活性。在 PBMC 中测试的 29 个主要 HIV-1 分离株中，发现 TAF 的平均 EC50 为 3.5 nM，而内部对照 AZT 的平均 EC50 为 11.8 nM。对于 HIV-2 分离株，AZT 的平均 EC50 为 6.4 nM，TAF 的平均 EC50 为 1.8 nM [2]。

替诺福韦的酰胺化前药称为 GS-7340 富马酸盐，或替诺福韦艾拉酚胺富马酸盐。与富马酸替诺福韦二吡呋酯（TDF）相比，它具有更好的血浆稳定性和良好的口服生物利用度[1]。

肠道和肝脏S9稳定性[1]
GS-7340与狗或人的肠道和肝脏S9组分在37°C下以96孔板型在10μM下孵育120分钟。在添加化合物后的指定时间点，用9倍体积的含有25%乙腈和50%甲醇的水溶液淬灭样品。将板在3000g下离心30分钟，通过LC-MS/MS分析10μL所得溶液。数据（分析物与内标峰面积比）以半对数标度绘制，并使用指数拟合进行拟合。假设一级动力学，确定了半衰期和代谢速率。使用充分搅拌的肝脏清除模型，通过报告的方法根据半衰期计算预测的肝脏提取量。[1]

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血浆稳定性[1]
GS-7340在37°C下在狗或人血浆中以2μM孵育4小时。在指定的时间点，通过加入9倍体积的100%乙腈和内标来淬灭孵育的等分试样。最后一次收集后，将样品在3000g下离心30分钟，将上清液转移到含有等体积水的新板上，通过液相色谱法结合三重四极杆质谱法（LC-MS/MS）进行分析。

Caco-2渗透性[1]
如前所述，使用接种在12孔板中的人结肠癌细胞系caco-2的融合单层进行了双向渗透性研究。研究了浓度（10、100或1000μM）或外排转运体抑制对GS-7340渗透的影响。在运输缓冲液中用10μM环孢菌素a（CsA）预孵育细胞单层30分钟以使转运蛋白结合位点饱和后，评估了包括P-糖蛋白（Pgp）在内的外排转运蛋白的抑制效果。预孵育后，加入含有CsA和GS-7340的新鲜测定缓冲液，开始测定。每次测定均进行两次，并测定对照化合物（阿替洛尔、普萘洛尔和地高辛）的渗透性，以满足每批测定板的验收标准。

Animals[1]
Male beagle dogs between the ages of 6 and 18 months were used for the in life portion of this study. The animals were housed in accordance with the standards of the American Association for Accreditation of Laboratory Animal Care and were receiving a standard commercial diet. Animals were handled in strict accordance with the Guide for the Care and Use of Laboratory Animals, and the protocol was reviewed by the Institutional Animal Care and Use Committee at SRI International. Animals were between 7 and 11 kg at dosing.[1]

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Drug Administration[1]
For intravenous administration, GS-7340 was formulated in 5% dextrose in water. For oral administration, GS-7340 was formulated in 50 mM citric acid (pH 5.0) at doses of 2 to 10 mg/kg. For the 20 mg/kg dose, GS-7340 was formulated in 50 mM citric acid (pH 5.5) with 0.1% Polysorbate 20. To test the effect of efflux transport inhibition, dogs were administered 2 mg/kg GS-7340 1 h following pretreatment with a 75 mg capsule of CsA.[1]

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Plasma and PBMC Sample Collection[1]
Blood (approximately 0.5 mL) was collected at specified time points over 24 h from the jugular vein (intraportal vein infusion and oral pharmacokinetic studies), the jugular and portal veins (oral administration to portal vein cannulated dogs), or the cephalic vein (jugular vein infusion). Plasma was isolated in Vacutainer tubes containing EDTA as an anticoagulant by centrifugation. At select time points (1, 4, 8, and 24 h postdose) in the 5 mg/kg oral pharmacokinetic study, 8 to 10 mL of blood was collected into Vacutainer cell preparation tubes (Becton Dickinson) for isolation of peripheral blood mononuclear cells (PBMC) and processed following manufacturer instructions. Small aliquots (10 μL) of isolated PBMC pellets diluted in 0.9% NaCl were maintained at room temperature and used to determine cell count. 500 μL of 70% methanol was added to the remaining PBMC pellets and, together with plasma samples, stored at −80 °C until shipment for further processing and analysis.

[1]. Mechanism for effective lymphoid cell and tissue loading following oral administration of nucleotide prodrug GS-7340. Mol Pharm. 2013 Feb 4;10(2):459-66.

[2]. Antiviral activity, safety, and pharmacokinetics/pharmacodynamics of tenofovir alafenamide as 10-day monotherapy in HIV-1-positive adults. J Acquir Immune Defic Syndr. 2013 Aug 1;63(4):449-55.

Tenofovir alafenamide is an antiviral prescription medicine approved by the U.S. Food and Drug Administration (FDA) for the treatment of chronic hepatitis B virus infection (HBV) in adults and children 6 years of age and older who weigh at least 55 lb (25 kg) and who meet certain requirements, as determined by a health care provider.
HBV can be an opportunistic infection (OI) of HIV.

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Drug Indication
Vemlidy is indicated for the treatment of chronic hepatitis B (CHB) in adults and paediatric patients 6 years of age and older weighing at least 25 kg (see section 5. 1).

C25H33N6O9P

593.55

592.204

C, 50.68; H, 5.61; N, 14.18; O, 24.30; P, 5.23

379270-38-9

Tenofovir alafenamide;379270-37-8; 379270-38-9 (fumarate); 1392275-56-7 (hemifumarate);

68516365

White to off-white solid powder

3.656

227.89

4

14

14

41

799

3

[P@](C([H])([H])O[C@]([H])(C([H])([H])[H])C([H])([H])N1C([H])=NC2=C(N([H])[H])N=C([H])N=C12)(N([H])[C@]([H])(C(=O)OC([H])(C([H])([H])[H])C([H])([H])[H])C([H])([H])[H])(=O)OC1C([H])=C([H])C([H])=C([H])C=1[H].O([H])C(/C(/[H])=C(\[H])/C(=O)O[H])=O

MEJAFWXKUKMUIR-WIUYAKJJSA-N

InChI=1S/C21H29N6O5P.C4H4O4/c1-14(2)31-21(28)16(4)26-33(29,32-17-8-6-5-7-9-17)13-30-15(3)10-27-12-25-18-19(22)23-11-24-20(18)27;5-3(6)1-2-4(7)8/h5-9,11-12,14-16H,10,13H2,1-4H3,(H,26,29)(H2,22,23,24);1-2H,(H,5,6)(H,7,8)/b;2-1+/t15-,16+,33?;/m1./s1

(S)-isopropyl 2-(((S)-((((R)-1-(6-amino-9H-purin-9-yl)propan-2-yl)oxy)methyl)(phenoxy)phosphoryl)amino)propanoate fumarate

GS734; GS-734; GS 734; GS 7340; GS-7340; GS7340; TAF; Tenofovir alafenamide fumarate; GS-7340 fumarate; 379270-38-9; GS-7340 monofumarate; H2S5S51WW6; GS-7340 (fumarate); GS-7339 monofumarate; L-Alanine, N-((S)-(((1R)-2-(6-amino-9H-purin-9-yl)-1-methylethoxy)methyl)phenoxyphosphinyl)-, 1-methylethyl ester, (2E)-2-butenedioate (1:1); trade name: Genvoya.

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 : ≥ 36 mg/mL (~60.76 mM)
H2O : ≥ 25 mg/mL (~42.19 mM)

配方 1 中的溶解度: 6.67 mg/mL (11.26 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶。 (<60°C).

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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：
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10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
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