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| Targets |
The primary molecular target of Trecovirsen sodium is the gag gene mRNA of HIV-1. Specifically, it binds to a 25-nucleotide sequence within the gag coding region through Watson-Crick base pairing. This binding is highly sequence-specific, with the antisense oligonucleotide hybridizing to the complementary RNA sequence. Once bound, the heteroduplex serves as a substrate for RNase H, an endogenous cellular enzyme that cleaves the RNA strand of DNA-RNA hybrids. This results in degradation of the gag mRNA, preventing translation of gag polyprotein precursors (p55, p41, p24). The compound does not directly target viral enzymes (reverse transcriptase, protease, integrase) but rather blocks viral protein synthesis. It is not a receptor ligand but a nucleic acid-based therapeutic .
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| ln Vitro |
In vitro studies demonstrate that Trecovirsen sodium inhibits HIV-1 replication in infected cell cultures. Using human T-lymphoid cell lines (e.g., MT-2, MT-4, H9) or peripheral blood mononuclear cells (PBMCs) infected with laboratory-adapted HIV-1 strains (e.g., IIIB, RF), the compound reduces p24 antigen production and syncytium formation. The IC50 values range from 0.1 to 1 microM depending on the cell type and viral strain. At concentrations up to 10 microM, no significant cytotoxicity is observed in uninfected cells as measured by MTT assays or trypan blue exclusion. The compound shows sequence-specific activity, as mismatched oligonucleotides do not inhibit viral replication. It is effective against multiple HIV-1 isolates but is less active against HIV-2 due to sequence divergence in the gag gene. The compound also inhibits HIV-1 reverse transcriptase in cell-free assays, though this is not the primary mechanism .
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| ln Vivo |
In vivo antiviral activity of Trecovirsen sodium has been evaluated in murine models using HIV-1-infected human PBMC reconstitution (SCID-hu or hu-PBL-SCID mice). In these models, intravenous administration at doses of 5-25 mg/kg/day for 2-4 weeks reduces plasma viral load as measured by HIV-1 RNA (RT-PCR) and p24 antigen levels. Splenic and lymph node viral titers are also decreased compared to vehicle controls. CD4+ T-cell counts are partially preserved in treated animals. However, the compound has limited activity in standard mouse models because HIV does not replicate in murine cells, necessitating humanized mouse models. No clinical efficacy data are available from animal studies beyond these model systems. In ex vivo studies, PBMCs from treated animals show reduced HIV recovery upon stimulation .
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| Enzyme Assay |
For RNase H-dependent antisense activity assays, a cell-free system is used. A 5'- or 3'-radiolabeled (32P) HIV-1 gag RNA transcript (target) is synthesized by in vitro transcription. Trecovirsen (1-100 nM) is mixed with the labeled RNA (10-50 pM) in hybridization buffer (10 mM Tris-HCl pH 7.5, 100 mM KCl, 5 mM MgCl2, 1 mM DTT) and heated to 65degC for 5 minutes, then cooled to 37degC over 30 minutes. RNase H (0.1-1 U) is added and incubated for 15-60 minutes at 37degC. Reactions are stopped with formamide loading dye, heated to 95degC, and resolved on a 6-15% denaturing polyacrylamide gel. Cleavage products are visualized by autoradiography or phosphorimaging. The RNA cleavage pattern indicates specific target recognition. Alternatively, a fluorescently labeled RNA (FAM or Cy5) can be used with gel scanning .
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| Cell Assay |
In vitro antiviral activity is assessed using HIV-1-infected human T-cell lines or PBMCs. MT-2 or MT-4 cells are cultured in RPMI-1640 medium with 10% FBS, 2 mM L-glutamine, 100 U/mL penicillin, and 100 microg/mL streptomycin at 37degC in 5% CO2. Cells are infected with HIV-1 at a multiplicity of infection (MOI) of 0.01-0.1 for 2 hours, washed, and then cultured with various concentrations of Trecovirsen (0.01-10 microM) for 5-7 days. Controls include scrambled oligonucleotide and no treatment. Supernatants are collected every 2-3 days for p24 antigen ELISA (HIV-1 p24 kit). Cytotoxicity is assessed in parallel using uninfected cells with MTT or CellTiter-Glo assays. The 50% inhibitory concentration (IC₅0) is calculated, along with the 50% cytotoxic concentration (CC₅0). The selectivity index (CC₅0/IC₅0) is typically >10 .
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| Animal Protocol |
In vivo efficacy is evaluated in the hu-PBL-SCID mouse model. SCID (severe combined immunodeficient) mice are engrafted with human PBMCs (20-30 × 10⁶ cells/mouse, intraperitoneally) to reconstitute a human immune system. After 2-4 weeks, mice are infected with HIV-1 (e.g., IIIB or JR-CSF strain, 1,000 TCID₅0) intraperitoneally. Trecovirsen sodium is administered intravenously via tail vein injection at 5-25 mg/kg/day for 14-28 days, either as daily bolus or continuous infusion (using osmotic pumps). Control groups receive saline or scrambled oligonucleotide. Plasma HIV-1 RNA is quantified by RT-PCR. At sacrifice, peritoneal lavage cells and spleens are harvested for p24 antigen measurement and proviral DNA detection (Alu-gag PCR). CD4/CD8 ratios are determined by flow cytometry of human cells (hCD45+/hCD3+). Endpoint: reduction in viral load ≥1 log .
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| ADME/Pharmacokinetics |
Pharmacokinetic studies in HIV-positive subjects and healthy volunteers show that Trecovirsen sodium after intravenous infusion has a biphasic elimination profile. The initial distribution half-life (t1/2alpha) is approximately 0.5-1 hour, and the terminal elimination half-life (t1/2beta) is 10-20 hours. Peak plasma concentrations (Cmax) are dose-proportional over 0.1-2 mg/kg. Plasma protein binding is high (>90%) due to the phosphorothioate backbone. The compound distributes extensively to tissues, particularly liver, kidney, and lymph nodes. It undergoes limited metabolism via exonucleases, releasing shorter oligonucleotide fragments. Renal excretion accounts for approximately 30-50% of the dose as intact and degraded product. No cytochrome P450-mediated metabolism occurs. Pharmacokinetic variability is moderate (CV 20-40%). The compound does not accumulate significantly with daily dosing .
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| Toxicity/Toxicokinetics |
Toxicology studies in rats and monkeys have been conducted. In rats, repeated intravenous doses of 10 mg/kg/day for 28 days are generally well-tolerated, with mild increases in liver transaminases (ALT, AST) and mild thrombocytopenia (decreased platelet count). At higher doses (≥30 mg/kg/day), more pronounced thrombocytopenia, prolongation of activated partial thromboplastin time (aPTT), and complement activation (elevated CH50, Bb, SC5b-9) are observed. Renal effects (proteinuria, increased BUN) occur at high doses. In monkeys, dose-limiting toxicity is also thrombocytopenia (dose-dependent, reversible). No genotoxicity is observed in Ames or micronucleus assays. No carcinogenicity studies have been reported. The compound has been tested in HIV-positive subjects in Phase I/II trials, with adverse events including mild injection site reactions, thrombocytopenia (dose-limiting), and fatigue. No organ toxicity requiring clinical intervention at doses ≤2 mg/kg/day .
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| References |
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| Additional Infomation |
Trecovirsen sodium (GEM 91) was investigated in clinical trials in the 1990s for the treatment of HIV infection but was not approved by the FDA or other regulatory agencies. Phase I/II studies in HIV-positive individuals showed modest antiviral activity (reduction in p24 antigen and viral RNA) with dose-limiting toxicity (thrombocytopenia). The maximum tolerated dose (MTD) was determined to be approximately 2 mg/kg/day as a continuous intravenous infusion. Oral bioavailability was negligible, so parenteral administration was required. Further development was discontinued due to the advent of more effective antiretroviral therapies (protease inhibitors, HAART) and the toxicity profile of first-generation antisense oligonucleotides. The compound remains a research tool for studying antisense mechanisms and phosphorothioate chemistry. No current clinical trials are active. It represents a historical candidate in the development of oligonucleotide therapeutics, having been among the first antisense compounds to enter human trials for infectious disease .
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| Appearance |
Solid powder
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| Synonyms |
GEM91 sodium
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
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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