| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| 500mg |
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| 1g |
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| Targets |
Broad-spectrum antiviral
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| ln Vitro |
The RNA-dependent RNA polymerase of the severe acute respiratory syndrome coronavirus 2 is an important target in current drug development efforts for the treatment of coronavirus disease 2019. Molnupiravir is a broad-spectrum antiviral that is an orally bioavailable prodrug of the nucleoside analogue β-D-N4-hydroxycytidine (NHC). Molnupiravir or NHC can increase G to A and C to U transition mutations in replicating coronaviruses. These increases in mutation frequencies can be linked to increases in antiviral effects; however, biochemical data of molnupiravir-induced mutagenesis have not been reported. Here we studied the effects of the active compound NHC 5’-triphosphate (NHC-TP) against the purified severe acute respiratory syndrome coronavirus 2 RNA-dependent RNA polymerase complex. The efficiency of incorporation of natural nucleotides over the efficiency of incorporation of NHC-TP into model RNA substrates followed the order GTP (12,841) > ATP (424) > UTP (171) > CTP (30), indicating that NHC-TP competes predominantly with CTP for incorporation. No significant inhibition of RNA synthesis was noted as a result of the incorporated monophosphate in the RNA primer strand. When embedded in the template strand, NHC-monophosphate supported the formation of both NHC:G and NHC:A base pairs with similar efficiencies. The extension of the NHC:G product was modestly inhibited, but higher nucleotide concentrations could overcome this blockage. In contrast, the NHC:A base pair led to the observed G to A (G:NHC:A) or C to U (C:G:NHC:A:U) mutations. Together, these biochemical data support a mechanism of action of molnupiravir that is primarily based on RNA mutagenesis mediated via the template strand [3].
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| ln Vivo |
Molnupiravir has strong antiviral properties and can stop SARS-CoV multiplication and illness [1]. It is administered orally every 12 hours for three days at a dose of 50–500 mg/kg. The duration of fever and viral load are dramatically reduced by molnupiravir (7 mg/kg), when administered orally twice daily for 3.5 days [2].
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| Enzyme Assay |
NTP incorporation and the effect of primer- or template-embedded NHC-MP on viral RNA synthesis[3]
NTP incorporation by SARS-CoV-2 RdRp and data acquisition and quantification were done as reported by us. Enzyme concentration was 100 or 200 nM for single and multiple nucleotide incorporation assays, respectively. RNA synthesis incubation time was 10 min. Data from single nucleotide incorporation assays were used to determine the preference for the natural nucleotide over NHC-TP. The selectivity value is calculated as a ratio of the incorporation efficiencies of the natural nucleotide over the nucleotide analogue. The efficiency of nucleotide incorporation is determined by the ratio of Michaelis–Menten constants Vmax over Km. The substrate for nucleotide incorporation is a 5-nt primer generated by incorporation of [α-32P]NTP into a 4-nt primer. Formation of the 5-nt primer is maximal at a given time point; however, its precise concentration is unknown. Hence, the product generated in the reaction is measured by quantifying the signal corresponding to the 6-nt primer product and dividing it to the total signal in the reaction (5-nt primer and 6-nt primer). This defines the product fraction. The product fraction is commonly multiplied by the total substrate concentration in order to determine the molar units of the Vmax, which is here not possible as explained above. Therefore, the unit of Vmax is reported as product fraction over time. The selectivity value is unitless as it is the ratio of two Vmax/Km measurements with the same units. RNA templates with embedded NHC-MP were produced as described by us. NHC-related protocol modifications are explained in Fig. S1.
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| Cell Assay |
Madin-Darby canine kidney (MDCK) cells (ATCC CCL-34) were grown at 37°C and 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 7.5% fetal bovine serum (FBS). Normal primary human bronchial tracheal epithelial cells (HBTECs) from a 30-year old healthy female donor were grown in BronchiaLife cell culture medium. These cells were obtained by the vendor under informed consent and adheres to the Declaration of Helsinki, The Human Tissue Act (UK), CFR Title 21, and HIPAA regulations. All regulatory approval lies with the vendor. Immortalized cell lines used in this study were routinely checked for microbial contamination (in approximately 6-month intervals). HBTECs were tested for microbial contamination on July 25, 2017 by LifeLine Cell Technology. Only HBTECs with a passage number 1-4 were used for this study [2].
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| Animal Protocol |
Animal/Disease Models: C57BL/6 mice (intranasal infection with SARS-CoV)[1]
Doses: 50, 150, 500 mg/kg Route of Administration: Oral; every 12 hrs (hours) for 3 days Experimental Results: Body weight loss is Dramatically diminished or prevented. Animal/Disease Models: Ca/09-infected female ferrets[1] Doses: 7 mg/kg Route of Administration: Oral; twice (two times) daily for 3.5 days Experimental Results: Shed virus load and duration of fever were Dramatically diminished. |
| References |
[1]. Sheahan TP, et al. An orally bioavailable broad-spectrum antiviral inhibits SARS-CoV-2 in human airway epithelial cell cultures and multiple coronaviruses in mice. Sci Transl Med. 2020 Apr 6. pii: eabb5883.
[2]. Toots M, et al. Characterization of orally efficacious influenza drug with high resistance barrier in ferrets and human airway epithelia. Sci Transl Med. 2019 Oct 23;11(515). pii: eaax5866. [3]. Molnupiravir promotes SARS-CoV-2 mutagenesis via the RNA template. Biol Chem. 2021 Jul; 297(1): 100770. |
| Molecular Formula |
C13H19N3O7
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|---|---|
| Molecular Weight |
329.31
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| Exact Mass |
329.12
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| Elemental Analysis |
C, 47.42; H, 5.82; N, 12.76; O, 34.01
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| CAS # |
2492423-29-5
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| Related CAS # |
Molnupiravir-d7
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| Appearance |
Typically exists as solids (or liquids in special cases) at room temperature
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| LogP |
-0.8
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| tPSA |
141Ų
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| SMILES |
O1[C@]([H])([C@@]([H])([C@@]([H])([C@@]1([H])C([H])([H])OC(C([H])(C([H])([H])[H])C([H])([H])[H])=O)O[H])O[H])N1C(N=C(C([H])=C1[H])N([H])O[H])=O
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| InChi Key |
O[C@@H]([C@H]([C@H](N1C(N/C(C=C1)=N/O)=O)O2)O)[C@H]2COC(C(C)C)=O
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| InChi Code |
HTNPEHXGEKVIHG-QCNRFFRDSA-N
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| Chemical Name |
InChI=1S/C13H19N3O7/c1-6(2)12(19)22-5-7-9(17)10(18)11(23-7)16-4-3-8(15-21)14-13(16)20/h3-4,6-7,9-11,17-18,21H,5H2,1-2H3,(H,14,15,20)/t7-,9-,10-,11-/m1/s1
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| Synonyms |
MK 4482; EIDD-2801; EIDD 2801; Molnupiravir; MK-4482; MK4482;
EIDD2801; prodrug-EIDD-1931; prodrug-EIDD 1931; prodrug-EIDD1931.
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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 : 50 mg/mL (151.83 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 12.05 mg/mL (36.59 mM) in 10% PEG400 2.5% Ethoxylated hydrogenated castor oil 87.5% water (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
Solubility in Formulation 2: ≥ 2.5 mg/mL (7.59 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 25.0 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (7.59 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. Solubility in Formulation 4: ≥ 2.5 mg/mL (7.59 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL corn oil and mix evenly. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.0367 mL | 15.1833 mL | 30.3665 mL | |
| 5 mM | 0.6073 mL | 3.0367 mL | 6.0733 mL | |
| 10 mM | 0.3037 mL | 1.5183 mL | 3.0367 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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