Antiviral; SARS-CoV-2; RSV; deuterated form of GS-621763

三异丁酸酯VV116还可以抑制RSV复制（EC50=1.20±0.32μM，CC50=95.92±9.27μM，SI=80，EC90=3.08±1.253μM），这表明VV116的酯部分易被细胞酶水解以释放母体核苷。这些化合物的抗RSV活性也在HEp-2和NHBE细胞中得到证实，HEp-2细胞和NHBE是RSV的其他许可细胞。[1]

---

药物化合物包括碳、氢和其他元素的稳定重同位素，主要作为影响药物开发过程中测量的示踪剂。药物的药代动力学和功能范围可能导致人们对突变的担忧[1]。延迟生成的化合物的潜在好处包括：（1）延迟生成的化合物可能能够扩展化合物的药代动力学特征，从而延长化合物的安全性、耐受性并改善耐受性； (2)延迟生成的化合物可以扩大肠道生物利用度。氘代化合物可能能够减少结肠和肠壁所需的首过代谢量，这将使更高比例的药物达到高生物利用度水平，这决定了其在低剂量下的疗效和更好的耐受性。 (3)增强新陈代谢功能。药物安全性、药物代谢 (4) 以及危险或反应性代谢物减少都是代谢物的潜在好处。氘代化学物质是无害的，并且有可能减轻或完全消除药物的负面影响。 (5)保存药用品质。根据早期的研究，氘代分子应保持与同类氢化合物相似的生物化学性质。

Mindeudesivir（VV116）（25、50和100 mg/kg；口服；每日两次，持续4天）显示出更强的活性，并将病毒滴度降至50 mg/kg的检测限以下，还可以减少呼吸道合胞病毒感染后的肺损伤[1]。VV116（25、50和100mg/kg；口服；单剂量）具有良好的PK特性和良好的安全性[1]。VV116（JT001）在Balb/c小鼠体内的药代动力学参数[1]。口服（25mg/kg）口服（50mg/kg）口服（100mg/kg）的Tmax（h）为0.42±0.14 0.42±0.12±0.14 Cmax（ng/mL）5360±560 11617±3443 24017±6521 AUC0-t（ng/mL·h）11461±1013 24594±1059 47799±6545 AUC0-∞（ng/mL-·h）11534±992 24739±1028 48014±6696 MRT0-∞（ng/mL·h）2.25±0.32 2.15±0.26 2.28±0.53 Tmax（小时）2.30±1.10 3.27±1.92 4.25±0.53动物模型：Balb/c小鼠[1]剂量：25、50和100mg/kg给药：口服；单剂量（药代动力学分析）结果：表现出良好的PK特性和良好的安全性。动物模型：Balb/c小鼠（6-8周；鼻内感染4×10^6 FFU的呼吸道合胞病毒）[1]剂量：25、50和100 mg/kg给药：口服；结果：在50mg/kg的剂量下，表现出更强的活性，病毒滴度降至检测限以下，也减少了呼吸道合胞病毒感染后的肺损伤。

---

考虑到Mindeudesivir（VV116）在体外抑制RSV的强效作用，我们在小鼠模型中进一步测试了VV16对RSV的影响。利巴韦林是临床上用于治疗呼吸道合胞病毒的标示外药物，被用作对照。为此，每只6-8周龄的Balb/c小鼠经鼻感染4×106 FFU的呼吸道合胞病毒（第0天），然后用VV116（25、50和100 mg/kg）或利巴韦林（50和100毫克/千克）双in die（b.i.d.）治疗（补充图S3）。我们之前的研究表明，在感染后第4天（p.i.），RSV感染的小鼠的病毒载量和病理学都达到了很高的水平，因此在这个时间点，小鼠被杀死，肺部被取出。用定量RT-PCR测量肺中的病毒RNA水平，用免疫斑点试验测量病毒颗粒载量（图1e）。值得注意的是，低剂量的VV116（25mg/kg）显示出与100mg/kg利巴韦林相当的抗病毒效果，分别将病毒RNA拷贝数和感染滴度降低了约1.5log10和约2.0log10（图1e）。中等剂量（50mg/kg）的VV116表现出更强的活性，并将病毒滴度降至检测限以下（图1e）。我们还通过组织化学分析评估了受攻击小鼠的肺部病理。呼吸道合胞病毒感染后，用赋形剂治疗的小鼠表现出严重的炎症，并出现肺泡炎性斑块。相比之下，在用VV116治疗的小鼠中只观察到轻微的肺浸润，表明VV116治疗可以减少呼吸道合胞病毒感染后的肺损伤[1]。

---

Balb/c小鼠的PK研究表明，在25至100mg/kg的剂量范围内，Mindeudesivir（VV116）具有线性PK曲线（图1c，补充表S6）。由于酯酶敏感前药的首过代谢，即使在100℃时，小鼠血浆中也未检测到VV116 口服给药后，母体核苷X1的血液浓度在0.5小时内迅速达到Cmax，在25mg/kg的剂量下，平均Cmax达到5360ng/ml（18.4µM，图1c，补充表S6、S7），远高于体外EC90值。X1的消除半衰期较短（2.3-4.25小时，补充表S6），支持每日两次给药方案。VV116的酯前药形式不仅旨在改善口服吸附，而且旨在规避核苷磷酰胺前药的肝脏靶向问题。临床前组织分布研究表明，X1在SD大鼠组织中广泛分布，5在Balb/c小鼠中也观察到X1的良好分布，X1在肺部的浓度约为肝脏的一半（图1d，补充表S8）。关于VV116的治疗窗口，大鼠14天重复给药口服毒性研究显示，无观测不良反应水平（NOAEL）为200mg/kg，此时X1的AUC0-t达到85151 ng h/ml（补充表S9），约为小鼠50mg/kg剂量的3.5倍。[1]

抗病毒活性和细胞毒性测定[1]
将A549、HEp-2或NHBE细胞接种到48孔板中并孵育过夜。在达到80%的细胞融合后，在细胞与不同浓度的药物孵育1小时后，将细胞感染RSV A2（A549中的MOI为2，HEp-2中的MOI0.5，NHBE细胞中的MOI/1）2小时。然后，去除病毒-药物混合物，用含药物的培养基培养细胞。接种后48小时，从细胞中提取总RNA，然后使用带有gDNA橡皮擦的PrimeScript RT试剂盒进行逆转录。为了确定病毒拷贝数，使用TB Green®Premix Ex TaqTM II进行了绝对定量RT-PCR。RSV A2 F片段用引物5'-CGAGCACAGAGAGAACTACCA-3'定量；以及5'-CCTTCTAGGTGCAGGACCTTA-3'.
在96孔板中进行细胞存活率测定，每种浓度取三份。所有药物在维持培养基（含2%FBS的DMEM）中以500微摩尔开始，用9个梯度稀释2倍。孵育48小时后，去除上清液，并在培养基中加入10μL WST-8（2-（2-甲氧基-4-（苯基）-3-（4-（苯基）-5（2,4-磺基苯）-2h-四唑鎓单钠盐）的维持培养基。孵育2小时后，使用分光光度计（BioTek）在450nm波长下测量板，并计算细胞存活率。

细胞活力测定
细胞类型： A549（感染 RSV）[1]
测试浓度： 0-1000 μM
孵育时间：48小时
实验结果：抑制A549细胞中RSV复制，EC50为1.20±0.32 μM，CC50为95.92。 ± 9.27 μM，选择性指数 (SI) 80。

---

细胞和病毒[1]
本研究中使用的所有细胞均在37℃、5%CO2的加湿培养箱中培养。人喉表皮样癌（HEp-2）细胞、Vero E6细胞和A549细胞在添加了10%胎牛血清的Dulbocco的Medified Eagle培养基（DMEM；Gibco）中生长。正常人支气管上皮（NHBE）细胞在支气管上皮细胞生长培养基（BEGM）中维持，并在BulletKit中提供所有补充剂。 RSV A2株在HEp-2细胞中生长。感染后3或4天，从感染细胞中收集病毒。简而言之，将RSV感染的细胞重复冷冻和解冻3次，然后在4℃下以1000rpm离心10分钟。然后，收集上清液并在-80℃下储存直至使用。如前所述，通过免疫斑点试验测定Vero E6细胞中RSV A2的病毒滴度1。所有RSV A2感染实验均在生物安全二级（BSL-2）实验室进行。

In vivo efficacy of Mindeudesivir (VV116) against RSV in mice [1]
Specific pathogen-free (SPF) female Balb/c mice at the age of 6–8 weeks were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The mice were housed in an SPF environment under standard conditions. All mouse experiments were approved by the ethics committee of the Wuhan Institute of Virology, Chinese Academy of Science (permit number WIVA25202113). Thirty mice (5 animals per group, 6 groups) were anaesthetized with isoflurane and challenged with 4×106 FFU of RSV A2 intranasally (i.n.). The mice were given drugs by intragastric administration. Treatments were commenced in 1 h post infection and continued for 4 days. Mice in the control group were given the solvent (40% PEG 400+10% HS 15+50% ultrapure water (v:v:v)). The mice were euthanized on the 4th day after challenge and their lungs were collected. The weight of the mice was recorded daily.
The left lung was fixed in tissue fixative solution, embedded, sectionized and stained with H&E to observe the pathological changes of lung tissue. After weighing the right lung, add 400μL PBS into the tube, grind it with a grinding instrument. One part of the grinding tissue was used to determine the virus titer, and the other part was used to determine virus copy number by extracted RNA from the tissue supernatant using viral DNA/RNA extraction Kit (TaKaRa, 9766). The determination of viral titers and subsequent treatment of the RNA obtained were the same as above.

---

Pharmacokinetic study of Mindeudesivir (VV116) in ICR mice, Balb/c mice, and SD rats [1]
ICR mice (N = 3 for each group, male) were fasted for 12 h before dosing (only for the oral administration). Mindeudesivir (VV116) dissolved in DMSO-enthanol-PEG300-saline (5/5/40/50, v/v/v/v) was administered intravenously at 5.0 mg/kg, and orally at 25 mg/kg. At 5 min, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 24 h post-dosing, blood samples were collected from the jugular vein or the submandibular vein into EDTA-K2 tubes, and immediately mixed with acetonitrile (20 µL blood + 80 μL acetonitrile). The concentrations of analytes in the blood were analyzed by LC-MS/MS.
A total of nine Balb/c mice (N = 3 for each group, male) were divided into three groups, and fasted for 12 h before dosing. The three groups received oral dose of Mindeudesivir (VV116) dissolved in 40%PEG400+10% Kolliphor® HS15+50% ultrapure water at 25 mg/kg, 50 mg/kg and 100 mg/kg, respectively. At 5 min, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 24 h post-dosing, blood samples were collected from the jugular vein or the submandibular vein into EDTA-K2 tubes, and immediately mixed with acetonitrile (20 µL blood + 80 μL acetonitrile). The concentrations of analytes in the blood were analyzed by LC-MS/MS.
SD rats (N = 3 for each group, male) were fasted for 12 h before dosing (only for the oral administration). The test compound (Mindeudesivir (VV116) or VV116-H) was administered intravenously at 5.0 mg/kg dissolved in DMSO-enthanol-PEG300-saline (5/5/40/50, v/v/v/v), and administered orally at 30 mg/kg dissolved in 40%PEG400+10% Kolliphor® HS15+50% ultrapure water. At 5 min, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 24 h post-dosing, blood samples were collected from the jugular vein into EDTA-K2 tubes. Serum samples were obtained following general procedures and the concentrations of analytes in the supernatant were analyzed by LC-MS/MS.

---

Tissue distribution study of Mindeudesivir (VV116) in Balb/c mice [1]
A total of thirty Balb/c mice were divided into five groups (3 animals/sex/group). VV116 was intragastrically administered at 100 mg/kg dissolved in 40%PEG400+10% Kolliphor® HS15+50%. At 0 (not administered), 0.25, 2, 6, and 24 h post-dosing, the five groups of mice were anesthetized, respectively. Blood samples were collected, and tissues including liver and lung were harvested. Tissue samples were individually homogenized, and blood samples were processed as above. The concentrations of X1 in liver, lung and blood were analyzed by LC-MS/MS.

---

Genetic toxicity assay [1]
The Ames test, the rat micronucleus assay, and the chromosome aberration test were conducted according to NMPA and ICH guidelines.
The Ames test was conducted to determine the mutagenicity of Mindeudesivir (VV116) using histidine-dependent Salmonella typhimurium (TA97a, TA98, TA100, TA1535) and tryptophan-dependent Escherichia coli (WP2). The experiment was carried out by plate permeating method under the -S9 non-metabolic and +S9 metabolic activation conditions. There were 6 dose groups for Mindeudesivir (VV116) (5, 50, 150, 500, 1500 and 5000 µg/dish under each condition) with the negative control (DMSO) and positive controls (ICR191, 2-nitrofluorene, sodium azide, 2-aminofluorene and methyl methanesulfonate). Under the conditions of -S9 and +S9, the average numbers of revertant colonies in the positive control group of each strain were at least twice that of the negative control group. The numbers of revertant colonies of each strain in all VV116 dose groups were less than twice that of the negative control group, and did not show dose-dependent increase. The result showed that VV116 was not mutagenic to histidine-dependent Salmonella typhimurium and tryptophan-dependent Escherichia coli.
The chromosome aberration test was conducted to evaluate whether Mindeudesivir (VV116) had the effect of inducing chromosome damage in Chinese hamster lung (CHL) cells by determining the aberration rate (excluding chromosome gap) under the -S9 and +S9 conditions. CHL cells were exposed to Mindeudesivir (VV116) without S9 for 4 h at the concentrations of 10, 20, 35, 40, 43, 45 and 48 μg/mL (-S9/4h group), or 24 h at the concentrations of 5, 10, 20, 25, 30, 35 and 40 μg/mL (-S9/24h group). In the presence of S9 mix, CHL cells were treated with VV116 for 4 h at the concentrations of 10, 25, 50,100 and 150 μg/mL (+S9/4h group). Meanwhile, negative (DMSO), and positive control groups (Mitomycin C and cyclophosphamide monohydrate) were set up. Based on the cytotoxicity of VV116, three doses of each group were chosen for chromosome aberration analysis. The positive compounds obviously induced chromosome aberrations compared with the negative control. For the -S9/4h group of VV116, the chromosome aberration rates at the concentrations of 20, 35 and 40 µg/mL were 0.0%, 0.3% and 0.0%, respectively; For the -S9/24h group, the rates at the concentrations of 10, 25 and 30 µg/mL were 1.0%, 0.3% and 0.3%, respectively. And for the +S9/4h group, the rates at the concentrations of 20, 50 and 150 µg/mL were 1.3%, 0.7% and 0.3%, respectively. The chromosome aberration rates of all Mindeudesivir (VV116) groups were within the background range, and showed no statistical difference compared with that of the negative control group. The result indicated that VV116 had no effect of inducing chromosome aberration in CHL cells.
The micronucleus assay in rats was conducted to evaluate whether Mindeudesivir (VV116) has the effect of inducing any increase of micronucleated polychromatic erythrocytes in rat bone marrow. Groups of male and female SD rats (5 animals/sex/group) received oral doses of VV116 at 0 (vehicle control), 100 (low), 200 (mild) and 500 mg/kg/d (high) for 14 days. The animals were sacrificed within 24 h after the last dose. Bone marrow smears were prepared for examining the ratio of polychromatic erythrocyte/(polychromatic erythrocyte + normochromatic erythrocyte) (PCE/(PCE + NCE)) and the micronucleus rate of polychromatic erythrocytes (MnPCE/PCE). The result showed that the PCE/(PCE + NCE) ratios of the female animals of the vehicle group, the low, the mild, and the high dose VV116 group were 0.65, 0.57, 0.58 and 0.58, respectively. For the male animals, the ratios were 0.62, 0.64, 0.66 and 0.60, respectively. VV116 did not show obvious bone marrow toxicity in rats. The assay was valid as the average micronucleus rates were 1.4‰ and 0.7‰ for the female and male rats in the vehicle group, respectively, which were within the historical range. The micronucleus rates of the female animals of the three VV116 groups were 1.2‰, 1.0‰ and 0.7‰, respectively, and for the male animals, the rates were 0.3‰, 0.7‰ and 0.3‰, respectively. There was no effect of any dose of VV116 on the micronucleus rate compared to the negative control. VV116 did not have the effect of inducing the increase of micronucleated polychromatic erythrocytes in rat bone marrow up to 500 mg/kg/d for 14 days.

---

Toxicokinetics of Mindeudesivir (VV116) in SD rats [1]
Groups of male and female SD rats (4 animals/sex/group) received repeated oral doses of Mindeudesivir (VV116) (dissolved in 40%PEG400+10% Kolliphor® HS15+50% ultrapure water) at 100 (low), 200 (mild) and 500 mg/kg/d (high) for 14 days. At day 1 and day 14, blood samples were collected from the jugular vein into EDTA-K2 tubes at various time points post-dose. Plasma samples were obtained following general procedures and the concentrations of analytes in the samples were analyzed by LC-MS/MS.

VV116 (JT001) is an oral nucleoside analogue candidate for SARS-CoV-2. Three Phase I studies aimed to evaluate the safety, tolerability, and pharmacokinetics of single and multiple escalation oral doses of VV116 in healthy subjects, as well as the effect of food on the pharmacokinetics and safety of VV116. The three studies were initiated sequentially: Study 1 (Single-Ascendance Study, SAD), Study 2 (Multiple-Ascendance Study, MAD), and Study 3 (Food Effects Study, FE). A total of 86 healthy subjects were enrolled. Subjects took either VV116 tablets or a placebo as instructed. Blood samples were collected at predetermined time points for pharmacokinetic analysis. The VV116 metabolite 116-N1 was detected in plasma, and its concentration was calculated for the calculation of pharmacokinetic parameters. In the single-dose (SAD) range of 25–800 mg, AUC and Cmax increased approximately dose-proportionately. T1/2 was within 4.80–6.95 hours. In multiple-dose administration (MAD), the cumulative ratios of Cmax and AUC indicated a slight accumulation of the drug after repeated administration of VV116. In standard meal administration (FE), the standard meal had no effect on the Cmax and AUC of VV116. No serious adverse events occurred in the study, and no subjects withdrew from the study due to adverse events. Therefore, VV116 demonstrated satisfactory safety and tolerability in healthy subjects, which supports continued investigation of VV116 in COVID-19 patients. [2]

---

Pharmacokinetic studies in Balb/c mice showed that Mindeudesivir (VV116) had linear pharmacokinetic characteristics in the dose range of 25 to 100 mg/kg (Figure 1c, Supplementary Table S6). Due to the first-pass metabolism of the esterase-sensitive prodrug VV116, it was not detected in mouse plasma even at doses up to 100 mg/kg. Following oral administration, the plasma concentration of the parent nucleoside X1 rapidly reached Cmax within 0.5 hours, with a mean Cmax of 5360 ng/ml (18.4 µM, Fig. 1c, Supplementary Tables S6 and S7) at a dose of 25 mg/kg, significantly higher than the in vitro EC90 value. The short elimination half-life of X1 (2.3–4.25 h, Supplementary Table S6) supports a twice-daily dosing regimen. The VV116 ester prodrug was designed not only to improve oral absorption but also to circumvent the liver-targeting issues associated with nucleoside phosphoramide prodrugs. Preclinical tissue distribution studies showed that X1 was widely distributed in SD rat tissues⁵, and good distribution was also observed in Balb/c mice, with lung X1 concentrations approximately half that in the liver (Fig. 1d, Supplementary Table S8). Regarding the therapeutic window of VV116, a 14-day repeated-dose oral toxicity study in rats showed that its NOAEL (no adverse reaction observed) was 200 mg/kg, at which point the AUC_0-t of X1 reached 85151 ng·h/ml (Supplementary Table S9), which is about 3.5 times that of the 50 mg/kg dose in mice^[1].

Safety [2]
No deaths, serious adverse events (SAEs), grade 3 or higher adverse events (AEs), or adverse events leading to drug discontinuation/interruption were reported in the three studies. All adverse events resolved without any treatment or intervention.
Study 1: Single-Elevation Dose Study
The number of subjects (incidence rates) of adverse events in the 25, 200, 400, 800, and 1200 mg dose groups and the placebo group were 2 (50%), 3 (50%), 3 (50%), 3 (50%), 0 (0%), and 5 (50%), respectively (Table 7). No dose-related adverse event occurrence was observed. In the single-escalation dose study, the incidence of adverse events was lower in subjects treated with Mindeudesivir (VV116) than in subjects treated with placebo (39.3% vs. 50%). Except for one case of grade 2 neutropenia, all adverse events were CTCAE grade 1 in severity. The dose escalation termination criteria were not met during the dose escalation period. The most common drug-related adverse events were sinus bradycardia, shortened PR interval on ECG, and elevated serum bilirubin.
Study 2: Multiple Dose Escalation Study
The number of subjects (incidence) of adverse events in the 200 mg, 400 mg, and 600 mg dose groups and the placebo group were 3 (33.3%), 5 (55.6%), 6 (66.7%), and 5 (55.6%), respectively (Table 8). The incidence of adverse events in subjects treated with Mindeudesivir (VV116) was comparable to that in subjects treated with placebo (51.9% vs. 55.6%). The occurrence of adverse events was dose-related. Except for one subject in the placebo group who experienced one of three cases of grade 2 nausea, the severity of adverse events was generally mild (CTCAE grade 1). The most common drug-related adverse events were elevated serum uric acid, dry mouth, urinary crystals, and nausea. Two subjects in the 400 mg dose group experienced grade 1 elevations in transaminases (elevated alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyltransferase), for a total of three cases. The elevations in transaminases were transient and resolved spontaneously.
Study 3: Food Effects Study
The number of subjects (incidence) of adverse events after fasting, after a standard meal, and after a high-fat meal were 0 (0%), 2 (16.7%), and 4 (33.3%), respectively. Two subjects experienced first-degree atrioventricular block after consuming a standard meal, while four subjects experienced positive urine bacteria test, urinary crystals, elevated blood pressure, and first-degree atrioventricular block after consuming a high-fat meal. All adverse events were grade CTCAE 1 in severity.
Other Safety Assessments
In Study 3, only one subject in the 400 mg dose group experienced a transient, mild increase in thyroid-stimulating hormone (TSH), which resolved spontaneously without treatment. No clinically significant abnormalities were found in sex hormone testing, ophthalmological examination, or thyroid ultrasound.

[1]. Oral remdesivir derivative VV116 is a potent inhibitor of respiratory syncytial virus with efficacy in mouse model. Signal Transduct Target Ther. 2022;7(1):123. Published 2022 Apr 16.

[2]. Safety, tolerability, and pharmacokinetics of VV116, an oral nucleoside analog against SARS-CoV-2, in Chinese healthy subjects [published online ahead of print, 2022 Mar 16]. Acta Pharmacol Sin. 2022;1-9.

Nucleoside antiviral drugs have a high genetic barrier to drug resistance because they target the highly conserved catalytic center of viral polymerase. VV116 has been shown to be effective against a variety of SARS-CoV-2 variants. Its good pharmacokinetic properties and safety make it a promising oral antiviral drug for the treatment of COVID-19. The in vivo efficacy in this study also provides strong evidence for the potential efficacy of VV116 in the treatment of RSV infection. Clinical studies of VV116 should be considered to alleviate RSV infection. [1] Mindrayvir (VV116) is a nucleoside analog prodrug used to treat COVID-19. Rifampin (RDV) was the first drug approved by the FDA for the treatment of COVID-19, and it is also a nucleoside analog. Compared with RDV, VV116 showed better in vitro antiviral activity and selectivity. In addition, VV116 can be administered orally and has good oral bioavailability, which is more convenient for COVID-19 patients than intravenous RDV. [2]
Mindexvir (VV116) is rapidly hydrolyzed to the metabolite 116-N1 after oral administration. 116-N1, not the parent drug VV116, was detected in plasma, and pharmacokinetic parameters were calculated based on this. Peak plasma concentrations of 116-N1 were rapidly reached after oral administration (median Tmax 1.00–2.50 h). In single-dose escalation studies, AUC and Cmax increased approximately dose-proportional across the dose range of 25–800 mg. However, no significant changes were observed when the dose increased from 800 mg to 1200 mg (AUC_0-t: 25886 vs. 28057 h·ng/mL; C_max: 2796 vs. 3086 ng/mL), suggesting that drug absorption may have reached saturation. Drug solubility is an important factor affecting drug absorption. When the drug reaches its maximum concentration (saturation solubility) at the absorption site, the drug absorption reaches its maximum value. It is speculated that the limited solubility of VV116 may be the reason for the drug absorption saturation. Within 0-72 hours after administration, the excretion fraction of 116-N1 in urine was 53.6%, while the excretion fraction of 116-N1 and VV116 in feces was 5.25%, indicating that VV116 is mainly excreted by the kidneys in the form of the metabolite 116-N1. [2] In the single escalation dose study, the mean half-life (t1/2) of Mindeudesivir (VV116) was 4.80–6.95 hours, suggesting that a twice-daily dosing regimen can be used for clinical treatment. Therefore, in the multiple escalation dose study, a regimen of twice-daily dosing (12 hours apart) was adopted for 5.5 days (days 1–6). The cumulative ratios of AUC and Cmax indicate that VV116 accumulates slightly after continuous administration. After multiple 200 mg doses on days 5 and 6, the trough concentration of 116-N1 was in the range of 242–345 ng/mL (Table 5), which is higher than the EC90 of 116-N1 against the omicron variant in preclinical anti-SARS-CoV-2 trials (186.5 ng/mL). Therefore, a dosing regimen of 200 mg or more twice daily can maintain effective antiviral drug concentrations and is recommended for use in subsequent clinical studies in COVID-19 patients. [2]
The median Tmax under fasting, standard meal, and high-fat meal conditions were 1.50, 3.00, and 2.50 hours, respectively, indicating that eating can prolong the time to peak concentration. Compared with fasting, the geometric mean ratio (GMR) of Cmax (90% CI) under both standard and high-fat meal conditions was in the range of 80%–125%; the GMR (90% CI) of AUC under standard meal conditions was also in the range of 80%–125%, but AUC0-t and AUC0-∞ increased slightly by 26.32% and 24.67% respectively under high-fat meal conditions. Since food intake has no effect on the Cmax of minoxidil (VV116), while a high-fat diet slightly increases AUC, it is recommended that VV116 be taken on an empty stomach or after a meal in the treatment of COVID-19. [2]
In single-dose escalation studies, no obvious dose-related trend was observed, and the proportion of subjects reporting adverse events in the placebo group (50.0%) was higher than that in the minoxidil (VV116) group (39.3%). Except for one case of grade 2 neutropenia, all adverse events were CTCAE grade 1. In multiple dose escalation studies, the incidence of adverse events in the VV116 group was comparable to that in the placebo group (51.9% vs. 55.6%). The incidence of adverse events was slightly dose-dependent. Only one subject in the 400 mg dose group reported elevated alanine aminotransferase (ALT) and aspartate aminotransferase (AST). All adverse events (AEs) in subjects treated with VV116 were grade 1 and resolved without treatment. No serious adverse events occurred during the study, and no subjects withdrew from the study due to adverse events. Preclinical animal toxicology studies have suggested that VV116 may be toxic to the eyes, thyroid, and gonads. In this study, we performed ophthalmological examinations, thyroid function tests, thyroid ultrasound examinations, and sex hormone tests on healthy subjects before and after VV116 administration. No significant toxicity was observed in these organs. Overall, VV116 demonstrated a satisfactory safety profile in healthy subjects across the three studies. [2]
Hepatotoxicity was the major adverse reaction (ADR) of RDV, manifested as elevated transaminases. In the Phase I clinical study (GS-US-399-5505 study), subjects received a loading dose of 200 mg of RDV followed by a dose of 100 mg for up to 9 days. Grade 1 or 2 transient ALT elevations were observed in 9 out of 20 subjects (45%). Elevated transaminases were also reported as the most common adverse reaction in COVID-19 patients treated with RDV. In the multiple-dose escalation study of mindiva (VV116), only 1 out of 27 subjects (3.7%) experienced a Grade 1 transient ALT elevation, which resolved spontaneously upon discontinuation of VV116. This is likely due to the high liver targeting of RDV, with a liver/blood concentration ratio approximately 21 times that of VV116. Four hours after a single intravenous injection of 10 mg/kg [14C]RDV, the liver/blood concentration ratio of RDV (calculated as 14C-GS-5734 equivalent) was 57.8; while two hours after a single oral administration of 30 mg/kg VV116, the liver/blood concentration ratio of VV116 (calculated as major metabolite 116-N1) was only 2.8. Although the risk of hepatotoxicity of VV116 is lower than that of RDV, liver function will continue to be monitored in a subsequent Phase II study of VV116 in COVID-19 patients. [2] Conclusion [2] Mindeudesivir (VV116) showed satisfactory safety and tolerability in healthy subjects. After oral administration of VV116, plasma drug concentrations of 116-N1 rapidly reached peak (median Tmax 1.00–2.50 hours). In the dose range of 25–800 mg, AUC and Cmax increased in approximately dose-proportional manner, while drug absorption may saturate at a dose of 800 mg. Standard meals had no effect on the Cmax and AUC of VV116. Effective antiviral concentrations were achieved with multiple doses, twice daily at dose levels of 200–600 mg. [2] In conclusion, the safety data and pharmacokinetic characteristics from these studies support continued investigation of VV116 in COVID-19 patients.

C24H32BRN5O7

583.450346946716

502.23

C, 49.41; H, 5.70; Br, 13.69; N, 12.00; O, 19.19

2779498-79-0

2647442-33-7; 2647442-13-3 (non-label form); 2779498-79-0 (HBr)

163409081

White to light yellow solid powder

168

2

11

11

37

887

4

Br.O1[C@H](COC(C(C)C)=O)[C@H]([C@H]([C@]1(C#N)C1=CC([2H])=C2C(N)=NC=NN12)OC(C(C)C)=O)OC(C(C)C)=O

HVRWXEFYGBTUNI-FTPFRNFUSA-N

InChI=1S/C24H31N5O7.BrH/c1-12(2)21(30)33-9-16-18(34-22(31)13(3)4)19(35-23(32)14(5)6)24(10-25,36-16)17-8-7-15-20(26)27-11-28-29(15)17;/h7-8,11-14,16,18-19H,9H2,1-6H3,(H2,26,27,28);1H/t16-,18-,19-,24+;/m1./s1/i7D;

[(2R,3R,4R,5R)-5-(4-amino-5-deuteriopyrrolo[2,1-f][1,2,4]triazin-7-yl)-5-cyano-3,4-bis(2-methylpropanoyloxy)oxolan-2-yl]methyl 2-methylpropanoate;hydrobromide

VV116; VV 116; VV116; 2779498-79-0; VV116 hydrobromide; Deuremidevir hydrobromide; P4L9Q2T94L; CHEMBL5314972; VV-116; JT001; JT-001; JT 001; Deuremidevir; Deuremidevir HBr; Deuremidevir hydrobromide; mindeudesivir; [GS621763; GS-621763; GS 621763]

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: 250 mg/mL (428.49 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (3.57 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。

---

配方 2 中的溶解度: ≥ 2.08 mg/mL (3.57 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。

---

View More

配方 3 中的溶解度: ≥ 2.08 mg/mL (3.57 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 储备液加入到 900 μL 玉米油中并混合均匀。

---

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。