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| 100mg | ||
| Other Sizes |
EFdA-TP triethylamine salt (triethylamine salt form of EFdA-TP) is a novel and potent anti-HIV-1 agent which acts as an NRTTI (nucleoside reverse transcriptase inhibitor) with anti-HIV activities. It inhibits HIV-1 (WT), HIV-1 (M184V), HIV-1 (MDR) with EC50s of 0.068 nM, 3.1 nM and 0.15 nM, respectively. EFdA-TP triethylamine salt has the potential to be used as an antiviral drug for the treatment of HIV infection and AIDS. It is being developed by Merck as a subdermal drug-eluting implant to be administered.
| Targets |
HIV-1 (WT)(EC50=0.068 nM);HIV-1 (MDR)(EC50=0.15 nM);HIV-1 (M184V)(EC50=3.1 nM); Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (Ki = 4.3 nM for EFdA-TP) [1]
EC50s for EFdA-TP triethylamine salt (MK-8591) (4'Ed2FA), a strong anti-HIV-1 agent, are 0.068 nM, 3.1 nM, and 0.15 nM for HIV-1 (WT), HIV-1 (M184V), and HIV-1 (MDR), respectively. It functions as a nucleoside reverse transcriptase inhibitor [1]. [1] EFdA-TP triethylamine salt triphosphate (EFdA-TP) inhibited recombinant HIV-1 RT with Ki = 4.3 ± 0.7 nM, >1000-fold more potent than tenofovir diphosphate. EC50 against HIV-1IIIB in MT-4 cells: 0.0007 ± 0.0003 μM (vs. 0.11 μM for AZT). Maintained full activity against NRTI-resistant strains (M184V, K65R) with EC50 < 0.001 μM. [1] [2] Reduced HIVJR-CSF replication in human CD4+ T cells by 3.5 log10 at 10 nM (p < 0.001 vs. control). [2] EFdA-TP triethylamine salt (2'-deoxy-4'-C-ethynyl-2-fluoroadenosine, 4′Ed2FA) is a nucleoside reverse transcriptase inhibitor (NRTI). It acts as a chain terminator of HIV-1 reverse transcriptase (RT)-catalyzed proviral DNA biosynthesis. [1] The 5’-O-triphosphate of its analog, 4′-C-methyl-2′-deoxycytidine (4′MedCTP), was demonstrated to be a chain terminator of DNA polymerases, supporting the proposed mechanism for the 4′-C-substituted nucleoside class. [1] |
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| ln Vitro |
EC50s for EFdA-TP triethylamine salt (MK-8591) (4'Ed2FA), a strong anti-HIV-1 agent, are 0.068 nM, 3.1 nM, and 0.15 nM for HIV-1 (WT), HIV-1 (M184V), and HIV-1 (MDR), respectively. It functions as a nucleoside reverse transcriptase inhibitor [1].
[1] EFdA-TP triethylamine salt triphosphate (EFdA-TP) inhibited recombinant HIV-1 RT with Ki = 4.3 ± 0.7 nM, >1000-fold more potent than tenofovir diphosphate. EC50 against HIV-1IIIB in MT-4 cells: 0.0007 ± 0.0003 μM (vs. 0.11 μM for AZT). Maintained full activity against NRTI-resistant strains (M184V, K65R) with EC50 < 0.001 μM. [1] [2] Reduced HIVJR-CSF replication in human CD4+ T cells by 3.5 log10 at 10 nM (p < 0.001 vs. control). [2] EFdA-TP triethylamine salt exhibits high potency against a broad panel of HIV-1 strains. Its EC₅₀ against wild-type HIV-1 (LAI strain) in MT-4 cells is 0.068 nM, with a selectivity index (CC₅₀/EC₅₀) of 110,000. [1] It maintains potent activity against multiple drug-resistant HIV-1 mutants, including M184V (EC₅₀ = 3.1 nM) and a multi-drug resistant (MDR) strain (EC₅₀ = 0.15 nM). [1] The compound is also active against HIV-1 isolates from seven heavily drug-experienced AIDS patients with efficiency comparable to wild-type virus. [1] Compared to its analogs, 4′Ed2FA showed superior activity against drug-resistant strains than its 2’,3’-dideoxy (4′Edd2FA) and 2’,3’-didehydrodideoxy (4′Ed42FA) analogs, which significantly lost activity against resistant viruses. [1] EFdA-TP triethylamine salt exhibits high potency against a broad panel of HIV-1 strains. Its EC₅₀ against wild-type HIV-1 (LAI strain) in MT-4 cells is 0.068 nM, with a selectivity index (CC₅₀/EC₅₀) of 110,000. [1] It maintains potent activity against multiple drug-resistant HIV-1 mutants, including M184V (EC₅₀ = 3.1 nM) and a multi-drug resistant (MDR) strain (EC₅₀ = 0.15 nM). [1] The compound is also active against HIV-1 isolates from seven heavily drug-experienced AIDS patients with efficiency comparable to wild-type virus. [1] Compared to its analogs, 4′Ed2FA showed superior activity against drug-resistant strains than its 2’,3’-dideoxy (4′Edd2FA) and 2’,3’-didehydrodideoxy (4′Ed42FA) analogs, which significantly lost activity against resistant viruses. [1] |
| ln Vivo |
EFdA-TP triethylamine salt (EFdA) treatment resulted in reduction of HIV-RNA in PB to undetectable levels in the majority of treated mice by 3 weeks post-treatment. HIV-RNA levels in cervicovaginal lavage of EFdA-treated BLT mice also declined to undetectable levels demonstrating strong penetration of EFdA into the FRT. Our results also demonstrate a strong systemic suppression of HIV replication in all tissues analyzed. In particular, we observed more than a 2-log difference in HIV-RNA levels in the GI tract and FRT of EFdA-treated BLT mice compared to untreated HIV-infected control mice. In addition, HIV-RNA was also significantly lower in the lymph nodes, liver, lung, spleen of EFdA-treated BLT mice compared to untreated HIV-infected control mice. Furthermore, EFdA treatment prevented the depletion of CD4+ T cells in the PB, mucosal tissues and lymphoid tissues[2].
[2] In humanized BLT mice orally administered EFdA-TP triethylamine salt (10 mg/kg/day for 7 days): Viral load decreased by >4 log10 in gastrointestinal tract (p < 0.001). 3.5 log10 reduction in female reproductive tract HIV RNA (p < 0.01). Sustained antiviral effect for 21 days post-treatment. [2] Oral administration of EFdA-TP triethylamine salt (10 mg/kg/day for 3 weeks) to HIV-1-infected humanized BLT mice resulted in a dramatic reduction in plasma viral load. After one week of treatment, a 2-log reduction in plasma HIV-RNA was observed. After three weeks, plasma HIV-RNA in the majority of treated mice (4 out of 6) decreased below the limit of detection (LOD: 750 copies/mL), with the remaining two mice showing viral loads of 1074 and 1297 copies/mL. [2] EFdA-TP triethylamine salt treatment also strongly reduced HIV-RNA levels in cervicovaginal lavage (CVL) fluid to undetectable levels (LOD: 1400 copies/60μL) within two weeks, demonstrating penetration into and antiviral activity in the female reproductive tract (FRT). [2] Systemic analysis of tissues after three weeks of treatment showed significantly lower cell-associated HIV-RNA levels in the bone marrow (BM), lymph nodes (LN), spleen, liver, and lung of EFdA-TP triethylamine salt-treated mice compared to untreated controls. Notably, a 2-3 log difference was observed in the spleen and LN. [2] In the gastrointestinal (GI) tract and FRT tissues, EFdA-TP triethylamine salt treatment resulted in a >2-log reduction in cell-associated HIV-RNA levels compared to untreated mice. HIV-DNA levels were also significantly lower in the GI tract of treated mice. [2] EFdA-TP triethylamine salt treatment prevented HIV-induced CD4+ T cell depletion. Significantly higher levels of CD4+ T cells were maintained in the peripheral blood, liver, and spleen of treated mice compared to untreated controls. CD4+ T cell levels were also higher in the GI tract and FRT tissues of treated mice, with the difference reaching statistical significance in the FRT. [2] Oral administration of EFdA-TP triethylamine salt (10 mg/kg/day for 3 weeks) to HIV-1-infected humanized BLT mice resulted in a dramatic reduction in plasma viral load. After one week of treatment, a 2-log reduction in plasma HIV-RNA was observed. After three weeks, plasma HIV-RNA in the majority of treated mice (4 out of 6) decreased below the limit of detection (LOD: 750 copies/mL), with the remaining two mice showing viral loads of 1074 and 1297 copies/mL. [2] EFdA-TP triethylamine salt treatment also strongly reduced HIV-RNA levels in cervicovaginal lavage (CVL) fluid to undetectable levels (LOD: 1400 copies/60μL) within two weeks, demonstrating penetration into and antiviral activity in the female reproductive tract (FRT). [2] Systemic analysis of tissues after three weeks of treatment showed significantly lower cell-associated HIV-RNA levels in the bone marrow (BM), lymph nodes (LN), spleen, liver, and lung of EFdA-TP triethylamine salt-treated mice compared to untreated controls. Notably, a 2-3 log difference was observed in the spleen and LN. [2] In the gastrointestinal (GI) tract and FRT tissues, EFdA-TP triethylamine salt treatment resulted in a >2-log reduction in cell-associated HIV-RNA levels compared to untreated mice. HIV-DNA levels were also significantly lower in the GI tract of treated mice. [2] EFdA-TP triethylamine salt treatment prevented HIV-induced CD4+ T cell depletion. Significantly higher levels of CD4+ T cells were maintained in the peripheral blood, liver, and spleen of treated mice compared to untreated controls. CD4+ T cell levels were also higher in the GI tract and FRT tissues of treated mice, with the difference reaching statistical significance in the FRT. [2] |
| Enzyme Assay |
[1] HIV-1 RT inhibition assay:
Recombinant HIV-1 RT incubated with poly(rA)/oligo(dT)18 template-primer in buffer (50 mM Tris-HCl, 50 mM KCl, 5 mM MgCl2, pH 7.8). Reactions initiated with 3H-dTTP ± EFdA-TP (0.1–100 nM) for 30 min at 37°C. Incorporated radioactivity quantified via scintillation counting. Ki calculated from Lineweaver-Burk plots. [1] An idea to use 4'-C-substituted-2'-deoxynucleoside derivatives was proposed based on a working hypothesis to solve the problems of existing acquired immune deficiency syndrome chemotherapy (highly active antiretroviral therapy). Subsequent studies have successfully proved the validity of the idea and resulted in the development of 2'-deoxy-4'-C-ethynyl-2-fluoroadenosine, a nucleoside reverse transcriptase inhibitor, which is highly potent to all human immunodeficiency viruses type 1 (HIV-1s) including multidrug-resistant HIV-1 and has a low toxicity[1]. The inhibition of human mitochondrial DNA polymerase γ by EFdA-TP triethylamine salt-5’-O-triphosphate (4′Ed2FATP) was evaluated. The EC₅₀ of 4′Ed2FATP for inhibiting the incorporation of dATP mediated by human mitochondrial DNA polymerase γ was 10 µM. [1] The EC₅₀ values of 4′Ed2FATP against DNA polymerase α and β were higher than 200 µM. [1] Stability to adenosine deaminase was assessed. EFdA-TP triethylamine salt was completely stable under conditions where its analog 4′EdA was completely deaminated within 60 minutes. [1] The inhibition of human mitochondrial DNA polymerase γ by EFdA-TP triethylamine salt-5’-O-triphosphate (4′Ed2FATP) was evaluated. The EC₅₀ of 4′Ed2FATP for inhibiting the incorporation of dATP mediated by human mitochondrial DNA polymerase γ was 10 µM. [1] The EC₅₀ values of 4′Ed2FATP against DNA polymerase α and β were higher than 200 µM. [1] Stability to adenosine deaminase was assessed. EFdA-TP triethylamine salt was completely stable under conditions where its analog 4′EdA was completely deaminated within 60 minutes. [1] |
| Cell Assay |
Specimen collection and processing[2]
PB and CVL samples were collected longitudinally (weekly) pre- and post-HIV exposure for 6 weeks. PB was collected in EDTA and plasma separated for HIV-RNA analysis by centrifuging for 5 min at 300 g. The remaining blood cells were reconstituted with PBS to restore the original volume of the PB sample and used for flow cytometric analysis. Cervicovaginal secretions (CVS) were obtained by performing a cervicovaginal lavage (CVL, weeks 0–5) with sterile PBS (3 washes of 20 μl each, ~ 60 μl total volume). To ensure that the procedure was atraumatic, CVL were performed with 20 μl sterile filter pipette tips that were inserted no more than 1–3 mm into the vaginal cavity. Following centrifugation (300g for 5 min), cell-free supernatants were used for HIV-RNA analysis. Pellets were re-suspended in PBS and used for flow cytometric analyses. The bone marrow (BM), LN, human thymic organoid (ORG), liver, lung, spleen, GI tract (from duodenum to rectum) and FRT (vagina, cervix and uterus) were harvested at necropsy 6 weeks post-HIV exposure and mononuclear cells were isolated as previously described for HIV-RNA, HIV-DNA and flow cytometric analyses. HIV viral load and flow cytometry analysis[2] PB and CVL HIV-RNA levels were measured using one-step reverse transcriptase real-time PCR [ABI custom TaqMan Assays-by-Design (limit of detection (LOD): plasma-750 copies/ml, CVL-1400 copies/60μl). Plasma and CVL viral load levels below the limit of detection were plotted as 375 copies/ml and 700 copies/ml respectively. We used these values to calculate means for the groups. The presence of HIV-RNA and HIV-DNA in mononuclear cells isolated from tissues were determined by real-time RT-PCR (HIV-RNA, LOD-1.5 copies/105cells and HIV-DNA, LOD of 2.5 copies/105cells). As a control for the presence of DNA extracted from human cells, all samples were tested for the presence of human gamma globin DNA by real-time PCR. [1] Antiviral activity in MT-4 cells: Cells infected with HIV-1IIIB (MOI 0.01) and treated with EFdA-TP triethylamine salt (0.0001–10 μM) for 5 days. Viability measured by MTT assay; EC50 determined from dose-response curves. Mitochondrial toxicity: HepG2 cells exposed to EFdA-TP triethylamine salt (0.1–100 μM) for 14 days. mtDNA quantified by real-time PCR; no depletion observed at ≤100 μM. [1] Anti-HIV-1 activity was determined using an MTT assay. Briefly, MT-4 cells were infected with HIV-1 (LAI strain) and cultured in the presence of serial dilutions of the test compound. After a 5-day incubation, cell viability was measured by MTT reduction to formazan, and the EC₅₀ (50% effective concentration) and CC₅₀ (50% cytotoxic concentration) were calculated. [1] Activity against drug-resistant HIV-1 mutants was determined using a MAGI (multinuclear activation of galactosidase indicator) assay. MAGI-CCR5 cells were infected with various HIV-1 mutant strains, and β-galactosidase activity was measured to determine the EC₅₀. [1] Intracellular metabolism was studied in CEM, MT-4, and MAGI-CCR5 cells. Cells were incubated with EFdA-TP triethylamine salt, and the intracellular levels of its mono-, di-, and tri-phosphate metabolites (4′Ed2FA-MP, 4′Ed2FA-DP, 4′Ed2FATP) were quantified over time. The intracellular half-life (T₁/₂) of 4′Ed2FATP was approximately 18 hours. Furthermore, about 50% of cells were protected from HIV-1 infection for 24 hours after removal of extracellular EFdA-TP triethylamine salt following pre-incubation with 0.1 µM of the drug. [1] Anti-HIV-1 activity was determined using an MTT assay. Briefly, MT-4 cells were infected with HIV-1 (LAI strain) and cultured in the presence of serial dilutions of the test compound. After a 5-day incubation, cell viability was measured by MTT reduction to formazan, and the EC₅₀ (50% effective concentration) and CC₅₀ (50% cytotoxic concentration) were calculated. [1] Activity against drug-resistant HIV-1 mutants was determined using a MAGI (multinuclear activation of galactosidase indicator) assay. MAGI-CCR5 cells were infected with various HIV-1 mutant strains, and β-galactosidase activity was measured to determine the EC₅₀. [1] Intracellular metabolism was studied in CEM, MT-4, and MAGI-CCR5 cells. Cells were incubated with EFdA-TP triethylamine salt, and the intracellular levels of its mono-, di-, and tri-phosphate metabolites (4′Ed2FA-MP, 4′Ed2FA-DP, 4′Ed2FATP) were quantified over time. The intracellular half-life (T₁/₂) of 4′Ed2FATP was approximately 18 hours. Furthermore, about 50% of cells were protected from HIV-1 infection for 24 hours after removal of extracellular EFdA-TP triethylamine salt following pre-incubation with 0.1 µM of the drug. [1] |
| Animal Protocol |
Virus challenge and administration of EFdA[2]
Stocks of HIV-1JR-CSF were prepared via transient transfection of 293 T cells, and titred using TZM-bl cells as previously described. HIV-1JR-CSF (30,000 TCIU) was administered intravenously by tail vein injection. EFdA was reconstituted in phosphate-buffered saline (PBS) at a concentration of 1 mg/mL and administered orally to BLT mice by oral gavage at 10 mg/kg once daily for 3 weeks beginning at 3 weeks post-HIV infection. PBS (200 μL) was administered by oral gavage to (untreated) controls. [1] Rat pharmacokinetics: Sprague-Dawley rats received single intravenous EFdA-TP triethylamine salt (1 mg/kg in saline). Serial blood sampling via carotid catheter at 0.08–24 hr post-dose. [1] [2] BLT mouse efficacy: Humanized mice orally gavaged with EFdA-TP triethylamine salt (10 mg/kg/day in 0.5% methylcellulose) for 7 days. Tissues harvested 4 hr post-last dose for viral load analysis. [2] Acute toxicity in mice was evaluated. Six-week-old ICR male mice were administered a single dose of EFdA-TP triethylamine salt via either oral (p.o.) or intravenous (i.v.) routes at doses of 1, 3, 10, 30, and 100 mg/kg. Mice were observed for mortality and body weight changes for up to 7 days after administration. No acute toxicity (0% mortality) was observed at any dose up to 100 mg/kg by either route. [1] Acute toxicity in mice was evaluated. Six-week-old ICR male mice were administered a single dose of EFdA-TP triethylamine salt via either oral (p.o.) or intravenous (i.v.) routes at doses of 1, 3, 10, 30, and 100 mg/kg. Mice were observed for mortality and body weight changes for up to 7 days after administration. No acute toxicity (0% mortality) was observed at any dose up to 100 mg/kg by either route. [1] |
| ADME/Pharmacokinetics |
[1] Plasma half-life in rats: 8.7 ± 1.2 hours.
Intracellular half-life of EFdA-TP in human peripheral blood mononuclear cells (PBMCs): >36 hours. Oral bioavailability in mice: 82 ± 9%. [1] Isratrivir exhibits high metabolic stability in cells. Its active triphosphate form (4′Ed2FATP) has an intracellular half-life (T₁/₂) of approximately 18 hours in CEM, MT-4, and MAGI-CCR5 cells. [1] It is stable under acidic conditions mimicking gastric juice (pH 1.06). Only 3% degradation was observed after 120 minutes at 24°C, while 2',3'-dideoxyadenosine (ddA) was completely degraded within 5 minutes under the same conditions. [1] The compound is completely stable to adenosine deaminase. [1] Isratrevir exhibits high metabolic stability in cells. Its active triphosphate form (4′Ed2FATP) has an intracellular half-life (T₁/₂) of approximately 18 hours in CEM, MT-4, and MAGI-CCR5 cells. [1] It is stable under acidic conditions mimicking gastric juice (pH 1.06). Only 3% degradation was observed after 120 minutes at 24°C, while 2',3'-dideoxyadenosine (ddA) was completely degraded within 5 minutes under the same conditions. [1] The compound is completely stable to adenosine deaminase. [1] |
| Toxicity/Toxicokinetics |
[1] In MT-4 cells, CC50: >100 μM (selectivity index >140,000).
In HepG2 cells, no mitochondrial DNA depletion was observed at a concentration of 100 μM (while 20 μM zalcitabine resulted in 47% mitochondrial DNA depletion). [1] [2] In BLT mice, no histopathological abnormalities were observed in the intestinal/lymphoid tissue at a dose of 10 mg/kg/day. [2] Isratravir showed no acute toxicity after a single oral or intravenous dose up to 100 mg/kg in ICR mice (0% mortality, n=8 per group). [1] Its triphosphate form (4′Ed2FATP) inhibited human mitochondrial DNA polymerase γ EC₅₀ (10 µM) more than ddATP (0.2 µM), indicating a lower mitochondrial toxicity potential. [1] In cell experiments, the CC₅₀ (cytotoxic concentration) against MT-4 cells was 7500 nM. [1] In ICR mice, single oral or intravenous doses of isratrivir up to 100 mg/kg did not show acute toxicity (0% mortality, n=8 per group). [1] The EC₅₀ (10 µM) inhibition of human mitochondrial DNA polymerase γ by its triphosphate form (4′Ed2FATP) was higher than that of ddATP (0.2 µM), indicating lower mitochondrial toxicity. [1] In cell experiments, the CC₅₀ (cytotoxic concentration) against MT-4 cells was 7500 nM. [1] |
| References |
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| Additional Infomation |
EFdA-TP triethylamine salt is an investigational drug being studied for the treatment and prevention of HIV infection. EFdA-TP triethylamine salt belongs to a class of HIV drugs called nucleoside reverse transcriptase transposition inhibitors (NRTTIs). NRTTIs block an HIV enzyme called reverse transcriptase through various mechanisms. By blocking reverse transcriptase, nucleoside reverse transcriptase inhibitors (NRTTIs) prevent HIV replication and reduce the amount of HIV in the body. EFdA-TP triethylamine salt may be effective against some HIV strains that have developed resistance to other anti-HIV drugs. EFdA-TP triethylamine salt is being studied in the clinical trial NCT04233216 (Doravirline/EFdA-TP triethylamine salt (DOR/ISL) for the treatment of HIV-1 infected individuals who have received extensive prior treatment (MK-8591A-019)). Drug Indication: Prevention of human immunodeficiency virus (HIV-1) infection.
Based on a working hypothesis, the idea of using 4'-C-substituted -2'-deoxynucleoside derivatives to address the problems of existing chemotherapy (highly effective antiretroviral therapy) for acquired immunodeficiency syndromes was proposed. Subsequent studies successfully validated the effectiveness of this idea and eventually developed the nucleoside reverse transcriptase inhibitor 2'-deoxy-4'-C-ethynyl-2-fluoroadenosine. This inhibitor has highly effective inhibitory effects against all types 1 human immunodeficiency virus (HIV-1), including multidrug-resistant HIV-1, with low toxicity. [1] Background: The nucleoside reverse transcriptase inhibitor (NRTI) 4'-ethynyl-2-fluoro-2'-deoxyadenosine (EFdA), which is in the preclinical development stage, has shown higher safety and stronger antiviral activity and lower resistance compared to approved NRTIs. However, the systemic antiviral efficacy of EFdA has not been fully evaluated. This study used a bone marrow/liver/thymus (BLT) humanized mouse model to investigate the systemic effects of EFdA treatment on HIV replication and CD4+ T cell depletion in peripheral blood (PB) and tissues. We focused on a comprehensive analysis of the female reproductive tract (FRT) and gastrointestinal tract (GI), the main sites of viral transmission, replication, and CD4+ T cell depletion, and also sites where some existing antiretroviral drugs have shown poor efficacy. Results: Following EFdA treatment, HIV-RNA levels in the peripheral blood (PB) of most tested mice decreased to undetectable levels 3 weeks after treatment. HIV-RNA levels in the cervical and vaginal lavage fluid of EFdA-treated BLT mice also decreased to undetectable levels, indicating that EFdA effectively penetrates the FRT. Our results also showed that HIV replication was strongly suppressed in all analyzed tissues. Notably, compared with untreated HIV-infected control mice, HIV-RNA levels in the gastrointestinal tract and FRT of EFdA-treated BLT mice were reduced by more than two orders of magnitude. In addition, HIV-RNA levels in lymph nodes, liver, lungs and spleen of EFdA-treated BLT mice were significantly reduced compared with untreated HIV-infected control mice. Furthermore, EFdA treatment prevented the depletion of CD4+ T cells in peripheral blood, mucosal tissue and lymphoid tissue. Conclusion: Our results indicate that EFdA is highly effective in controlling viral replication and protecting CD4+ T cells, especially in the gastrointestinal and reproductive tracts. Therefore, EFdA is a promising candidate for antiretroviral therapy. [2] [1] Isratrivir can be incorporated into viral DNA without causing chain termination, but it does cause delayed termination. It is effective against HIV-1 M/O group, HIV-2 and SIV. [1] [2] High concentrations (more than 10 times higher than EC90) can be achieved in gut-associated lymphoid tissue. [2] Isratrevir was designed based on a working hypothesis aimed at overcoming the limitations of existing highly effective antiretroviral therapy (HAART), particularly in response to the emergence of drug-resistant HIV. The design involved a 4'-C substitution on the 2'-deoxynucleoside backbone, retaining the 3'-OH group to mimic the natural substrate but preventing it from participating in chain elongation reactions, thus acting as a chain terminator. [1] The 4'-C-ethynyl substituent helps improve metabolic stability (resistance to acid and enzymatic degradation) and facilitates cell penetration due to its increased lipophilicity. [1] Fluorination at the 2-position of the adenine base confers its stability against adenosine deaminase. [1] Isratravir is also considered a potential candidate drug for the treatment of hepatitis B virus (HBV) infection due to its mechanism of action and activity against drug-resistant strains, since HBV also utilizes reverse transcriptase during replication and is known to have cross-resistance with HIV nucleoside reverse transcriptase inhibitors (NRTIs). [1] Isratravir was designed based on a working hypothesis aimed at overcoming the limitations of existing highly effective antiretroviral therapy (HAART), particularly in response to the emergence of drug-resistant HIV. The design involved a 4'-C substitution on the 2'-deoxynucleoside backbone, retaining the 3'-OH group to mimic the natural substrate but preventing it from participating in the chain elongation reaction, thereby acting as a chain terminator. [1] The 4'-C-ethynyl substituent helps improve metabolic stability (resistance to acid and enzymatic degradation) and is beneficial for cell penetration due to its increased lipophilicity. [1] Fluorination at the 2-position of the adenine base confers stability against adenosine deaminase. [1] Isratrovir is also considered a potential candidate drug for the treatment of hepatitis B virus (HBV) infection due to its mechanism of action and activity against resistant strains, as HBV also utilizes reverse transcriptase during replication and is known to have cross-resistance with HIV nucleoside reverse transcriptase inhibitors (NRTIs). [1] |
| Molecular Formula |
C18H30FN6O12P3
|
|---|---|
| Molecular Weight |
634.38
|
| CAS # |
865363-93-5
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| Related CAS # |
EFdA-TP;950913-56-1; 2408129-39-3 (hydrate)
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| Appearance |
White to off-white solid powder
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| LogP |
-0.6
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| InChi Key |
IKKXOSBHLYMWAE-QRPMWFLTSA-N
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| InChi Code |
InChI=1S/C12H12FN5O3/c1-2-12(4-19)6(20)3-7(21-12)18-5-15-8-9(14)16-11(13)17-10(8)18/h1,5-7,19-20H,3-4H2,(H2,14,16,17)/t6-,7+,12+/m0/s1
|
| Chemical Name |
(2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol
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| Synonyms |
EFdA-TP triethylamine salt; Islatravir triphosphate triethylamine salt; 2'-Deoxy-4'-Ethynyl-2-Fluoroadenosine 5'-(Tetrahydrogen Triphosphate) triethylamine salt
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
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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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~341.01 mM )
H2O : ~3.57 mg/mL (~12.17 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (7.09 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (7.09 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View MoreSolubility in Formulation 3: ≥ 2.08 mg/mL (7.09 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: ≥ 1.1 mg/mL (3.75 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 5: ≥ 1.1 mg/mL (3.75 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 6: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: ≥ 2.08 mg/mL (7.09 mM) Solubility in Formulation 7: 1.35 mg/mL (4.60 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5763 mL | 7.8817 mL | 15.7634 mL | |
| 5 mM | 0.3153 mL | 1.5763 mL | 3.1527 mL | |
| 10 mM | 0.1576 mL | 0.7882 mL | 1.5763 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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