PFV IN with the S217H alteration has an IC50 of 900 nM, making it ten times less sensitive to raltegravir. PFV IN exhibited 10% of WT's activity and was inhibited by Raltegravir at an IC50 of 200 nM, suggesting that PFV IN is less sensitive to IN strand transfer inhibitors (INSTIs) than WT IN is. Similar to the WT enzyme, S217Q PFV IN is also susceptible to raltegravir [1]. Glucuronidation, not the liver, is the mechanism of raltegravir metabolism. With a 95% inhibitory concentration of 31±20 nM in human T cell cultures, raltegravir demonstrates strong anti-HIV-1 action in vitro. Raltegravir exhibited anti-HIV-2 activity in CEMx174 cells as well, with an IC95 of 6 nM. Glucuronidation is the main mechanism of raltegravir metabolism. Strong glucuronidase UGT1A1 inducers should not be utilized since they drastically lower raltegravir concentrations. Hepatic cytochrome P450 activity is only slightly inhibited by raltegravir. Neither CYP3A4-dependent testosterone 6-beta-hydroxylase activity nor CYP3A4 RNA expression are induced by raltegravir [2]. Magnesium and calcium have been shown to decrease raltegravir's cellular permeability [3]. Effectively preventing viral replication is possible with raltegravir and related HIV-1 integrase (IN) strand transfer inhibitors (INSTIs) [4]. Latisavue successfully suppressed SIVmac251 replication in the acutely infected human lymphoid CD4+ T cell lines MT-4 and CEMx174, suggesting an EC90 in the low nanomolar range [5].

具有 S217H 改变的 PFV IN 的 IC50 为 900 nM，使其对拉替拉韦的敏感性降低十倍。 PFV IN 表现出 WT 10% 的活性，并被 Raltegravir 抑制，IC50 为 200 nM，表明 PFV IN 对 IN 链转移抑制剂 (INSTI) 的敏感性低于 WT IN。与 WT 酶类似，S217Q PFV IN 也对拉替拉韦敏感 [1]。拉替拉韦的代谢机制是葡萄糖醛酸化，而不是肝脏。拉替拉韦在人类 T 细胞培养物中的 95% 抑制浓度为 31±20 nM，在体外表现出强大的抗 HIV-1 作用。 Raltegravir 在 CEMx174 细胞中也表现出抗 HIV-2 活性，IC95 为 6 nM。葡萄糖醛酸化是拉替拉韦代谢的主要机制。不应使用强葡萄糖醛酸酶 UGT1A1 诱导剂，因为它们会大大降低拉替拉韦浓度。拉替拉韦仅轻微抑制肝细胞色素 P450 活性。拉替拉韦既不诱导 CYP3A4 依赖性睾酮 6-β-羟化酶活性，也不诱导 CYP3A4 RNA 表达 [2]。镁和钙已被证明可以降低拉替拉韦的细胞通透性[3]。拉替拉韦和相关的 HIV-1 整合酶 (IN) 链转移抑制剂 (INSTI) 可以有效防止病毒复制 [4]。 Latisavue 成功抑制了急性感染的人淋巴 CD4+ T 细胞系 MT-4 和 CEMx174 中的 SIVmac251 复制，表明 EC90 在低纳摩尔范围内 [5]。

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在体外实验中，Raltegravir 有效抑制野生型（WT）原型泡沫病毒（PFV）整合酶的链转移活性，其IC50为90 nM。[1]
携带S217H替换（对应于HIV-1的Q148H）的PFV整合酶对Raltegravir的敏感性降低，IC50约为900 nM，表明其效力相比野生型酶降低了10倍。[1]
携带S217Q替换的PFV整合酶对Raltegravir仍然敏感，IC50为40 nM。[1]
携带N224H替换（对应于HIV-1的N155H）的PFV整合酶对Raltegravir的敏感性也降低，IC50为200 nM，表明效力降低了约2倍。[1]

随着感染的进展，Ratetelevi 可改善感染 SIVmac251 的非人灵长类动物的病毒免疫状态。拉替拉韦单一疗法导致一种非人类灵长类动物的病毒载量检测不到[5]。

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在SIVmac251感染的猕猴（第1组）中，使用raltegravir（50或100 mg，每日两次随食物口服）进行为期十天的单药治疗，导致所有动物的病毒载量显著下降（P = 0.031）和CD4+ T细胞计数显著增加（P = 0.017）。100 mg组中的一只动物达到了检测不到的病毒载量（<40拷贝/mL）。[5]
从第11天开始，在raltegravir治疗方案（所有动物转为100 mg每日两次）中加入两种逆转录酶抑制剂——替诺福韦(PMPA, 20 mg/kg/天，皮下注射)和恩曲他滨(FTC, 50 mg/kg/天，皮下注射)后，病毒载量继续下降，所有动物在两周内达到检测不到的水平，并持续到随访结束（第52天）。CD4计数持续增加，恢复至接种前水平。[5]
在第二组SIVmac251感染的猕猴（第2组）中，raltegravir单药治疗（100 mg每日两次）七天，与治疗前历史值相比，也导致病毒载量显著下降，证实了可重复的抗病毒效果。一只在单药治疗后病毒载量检测不到的动物在治疗暂停后出现反弹。[5]
尽管联合治疗使病毒载量抑制到检测不到的水平，但PBMCs中的原病毒DNA水平保持稳定，治疗52天后没有显著变化，表明慢病毒储存库持续存在。[5]

进行了定量的PFV整合酶链转移实验。反应体系（40 µL）包含0.75 µM纯化的PFV整合酶、0.75 µM供体DNA（模拟预处理的病毒U5 DNA末端的双链寡核苷酸）、4 nM（300 ng）超螺旋质粒靶DNA、5 mM MgSO₄（作为二价金属辅因子）、4 µM ZnCl₂、125 mM NaCl、10 mM DTT、25 mM Bis-Tris propane-HCl缓冲液（pH 7.4）和0.8% DMSO。在指定浓度下将Raltegravir加入反应中。通过添加PFV整合酶启动反应，在37°C下孵育1小时，然后加入EDTA和SDS终止反应。对反应产物进行去蛋白化、乙醇沉淀，并在用溴化乙锭染色的1.5%琼脂糖凝胶上进行分离以进行可视化。对于定量，使用特异性引物和荧光DNA结合染料通过定量实时PCR测量整合产物。使用野生型PFV整合酶反应（无抑制剂）的连续稀释液生成的标准曲线来量化抑制剂处理样品中的整合效率。[1]

Caco-2单层渗透性实验: 将Caco-2细胞接种于聚碳酸酯膜Transwell上，培养21天形成单层。通过跨上皮电阻（TEER >600 Ω）和对[14C]甘露醇的低渗透性确认单层完整性。对于渗透性研究，改变顶端隔室的pH（使用如MES用于较低pH，HEPES/Tricine用于较高pH的缓冲液），而底外侧隔室保持pH 7.4。将raltegravir（用于pH效应研究为50 µM；用于金属/奥美拉唑研究为1 µM）加入顶端（用于顶端到底外侧运输）或底外侧（用于底外侧到顶端运输）隔室。平板在37°C、5% CO2下孵育。在不同时间点（例如0、30、60、90、120分钟）从接收隔室取样并用新鲜缓冲液替换。通过LC-MS/MS或闪烁计数分析raltegravir浓度。计算表观渗透性（Papp）和外排比。
细胞积累实验: 将Caco-2细胞接种于6孔板中，培养5天。洗涤细胞并用pH缓冲的孵育溶液（pH 5-8）平衡。加入raltegravir（1 µM），伴或不伴ABCB1抑制剂tariquidar（300 nM）的预孵育和共孵育。在37°C、5% CO2下孵育10分钟后，取细胞外样品，用冰缓冲液洗涤细胞，用水裂解。用乙腈处理细胞裂解物，离心，通过LC-MS/MS分析上清液中的raltegravir含量。[3]

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Indian rhesus macaques were mucosally inoculated (intrarectally or intravaginally) with 300 MID50 of pathogenic SIVmac251. Viral loads stabilized by week 12 post-infection. For drug treatment, animals were randomized to receive raltegravir monotherapy orally at 50 mg or 100 mg twice daily with food for 10 days. At day 11, animals originally on 50 mg were switched to 100 mg twice daily, and all animals received subcutaneous injections of tenofovir (PMPA, 20 mg/kg/day) and emtricitabine (FTC, 50 mg/kg/day) in addition to raltegravir. Treatment continued until day 52. Plasma samples were collected regularly for viral load measurement by quantitative RT-PCR. PBMCs were collected for proviral DNA quantification by PCR and for flow cytometric analysis of CD4+ T cell counts. Clinical chemistry and hematology were monitored. [5]

Raltegravir solubility is pH-dependent. At 10 mM, it was partly insoluble at pH 6.6 and below, but fully soluble at pH 6.8 to 8.
The lipophilicity (log P) of raltegravir, determined using octanol-water partition, decreased from ~1.06 to -1.29 as pH increased from 5 to 9.
The pKa of raltegravir was determined to be 6.7 using UV spectroscopy.
In vitro cellular permeability (apical-to-basolateral) of raltegravir across Caco-2 monolayers is significantly reduced with increasing extracellular pH (from 27.3 x 10^-6 cm/s at pH 5 to 2.9 x 10^-6 cm/s at pH 8.5).
Divalent cations (Mg2+, Ca2+) reduce the in vitro cellular permeability of raltegravir.
Clinical studies referenced show that co-administration with omeprazole (increasing gastric pH) increases raltegravir AUC and Cmax in healthy volunteers, and that antacids containing magnesium/aluminum reduce raltegravir C12.
The primary metabolic route of raltegravir is glucuronidation via UGT1A1. It is a weak substrate for drug transporters ABCB1, SLC22A6, and SLC15A1. [3]

Raltegravir is not a substrate or inhibitor of major cytochrome P450 enzymes.
Drug-drug interactions exist: Atazanavir (UGT1A1 inhibitor) increases raltegravir exposure, while rifampin (UGT1A1 inducer) decreases it.
Co-administration with antacids containing magnesium/aluminum resulted in 75% of subjects having a raltegravir C12 lower than the IC95 (15 ng/mL in 50% human serum). [3]

[1]. Molecular mechanisms of retroviral integrase inhibition and the evolution of viral resistance. Proc Natl Acad Sci U S A, 2010. 107(46): p. 20057-62.

[2]. Raltegravir: the first HIV type 1 integrase inhibitor. Clin Infect Dis. 2009 Apr 1;48(7):931-9.

[3]. Divalent metals and pH alter raltegravir disposition in vitro. Antimicrob Agents Chemother. 2012 Jun;56(6):3020-6.

[4]. Structural and functional analyses of the second-generation integrase strand transfer inhibitor dolutegravir (S/GSK1349572). Mol Pharmacol. 2011 Oct;80(4):565-72.

[5]. Response of a simian immunodeficiency virus (SIVmac251) to raltegravir: a basis for a new treatment for simian AIDS and an animal model for studying lentiviral persistence during antiretroviral therapy. Retrovirology, 2010. 7: p. 21.

Raltegravir (brand names: Isentress and Isentress HD) is a prescription medicine approved by the U.S. Food and Drug Administration (FDA) for the treatment of HIV infection in adults and children. One form of raltegravir, Isentress, is approved for adults and children weighing at least 4 lb and 4 oz (2 kg). Another form of raltegravir, Isentress HD, is approved for adults and children weighing at least 88 lb (40 kg). Raltegravir is always used in combination with other HIV medicines.
Raltegravir Potassium is the orally bioavailable potassium salt of a human immunodeficiency virus (HIV) integrase strand transfer inhibitor (HIV-1 INSTI) with HIV-1 antiviral activity. Raltegravir binds to and inhibits integrase, an HIV enzyme that inserts viral genetic material into the genetic material of the infected human cell. Inhibition of integrase prevents insertion of HIV DNA into the human DNA genome, thus blocking HIV replication.
A pyrrolidinone derivative and HIV INTEGRASE INHIBITOR that is used in combination with other ANTI-HIV AGENTS for the treatment of HIV INFECTION.
See also: Raltegravir (has active moiety); Lamivudine; raltegravir potassium (component of).

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Drug Indication
Isentress is indicated in combination with other anti-retroviral medicinal products for the treatment of human immunodeficiency virus (HIV 1) infection.

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Raltegravir is the first integrase strand transfer inhibitor (INSTI) approved for clinical use against HIV-1 infection. [1]
Crystal structures of the prototype foamy virus (PFV) intasome (integrase-DNA complex) bound to Raltegravir revealed its binding mode. Raltegravir binds to the active site of integrase, chelating the two catalytic divalent metal ions (Mg²⁺ or Mn²⁺) via a triad of heteroatoms in its pharmacophore. This binding displaces the 3’-terminal adenosine nucleotide of the viral DNA from the active site, thereby inactivating the intasome and blocking the strand transfer reaction. [1]
A key interaction involves a face-to-face π-π stacking between the oxadiazole ring of Raltegravir and the side chain of Tyr212 in PFV integrase (equivalent to Tyr143 in HIV-1 integrase). Substitutions at this position (e.g., Y143H/R/C in HIV-1) are known to confer resistance, which is explained by the loss of this stabilizing interaction. [1]
The study used PFV integrase as a model to understand resistance mutations in HIV-1 integrase. The S217H and N224H mutations in PFV integrase (analogous to Q148H and N155H in HIV-1) confer reduced susceptibility to Raltegravir. Crystal structures of mutant intasomes showed that these substitutions necessitate unfavorable conformational rearrangements in the active site to accommodate the inhibitor, explaining the mechanism of resistance. Specifically, the S217H (Q148H) mutation requires a significant backbone shift for inhibitor binding, while the N224H (N155H) mutation disrupts a stabilizing interaction between the asparagine and the DNA backbone that must be broken upon inhibitor binding. [1]
The presence of a serine at position 209 in PFV integrase (equivalent to Gly140 in HIV-1) likely explains why the single S217H mutant in PFV shows a high level of resistance similar to the HIV-1 Q148H/G140S double mutant. In the S217H mutant structure, Ser209 forms a hydrogen bond with His217, illustrating the potential interaction and co-evolution of residues at positions 140 and 148 in HIV-1 integrase under drug selection pressure. [1]

C20H20FKN6O5

482.51

482.111

871038-72-1

Raltegravir;518048-05-0;Raltegravir-d3 potassium;1246816-98-7;Raltegravir sodium;1292804-07-9

23668479

White to off-white solid powder

1.46 g/cm3

282ºC

2.131

155.07

2

9

6

33

843

0

IFUKBHBISRAZTF-UHFFFAOYSA-M

InChI=1S/C20H21FN6O5.K/c1-10-25-26-17(32-10)16(30)24-20(2,3)19-23-13(14(28)18(31)27(19)4)15(29)22-9-11-5-7-12(21)8-6-11;/h5-8,28H,9H2,1-4H3,(H,22,29)(H,24,30);/q;+1/p-1

potassium;4-[(4-fluorophenyl)methylcarbamoyl]-1-methyl-2-[2-[(5-methyl-1,3,4-oxadiazole-2-carbonyl)amino]propan-2-yl]-6-oxopyrimidin-5-olate

MK-0518; MK0518; MK 0518; MK-0518 potassium; Raltegravir; trade name: Isentress

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)

配方 1 中的溶解度: ≥ 2.08 mg/mL (4.31 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 20.8 mg/mL澄清DMSO储备液加入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: ≥ 2.08 mg/mL (4.31 mM) (饱和度未知) 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), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 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: ≥ 2.08 mg/mL (4.31 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 20.8 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 4 中的溶解度: 30% PEG400+0.5% Tween80+5% Propylene glycol : 30 mg/mL

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配方 5 中的溶解度: 25 mg/mL (51.81 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.

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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。