Viral polymerase-IN-1 HCl is a Gemcitabine analogue that effectively inhibits influenza A and B virus (influenza) infection with an IC90 value of 11.4-15.9 μM. Viral polymerase-IN-1 HCl has activity against SARS-CoV-2 infection. Viral polymerase-IN-1 HCl inhibits influenza virus infection by affecting viral RNA replication/transcription in cells.

Physicochemical Properties

Molecular Formula	C15H16CLF2N5O5
Molecular Weight	419.77
Exact Mass	419.08
CAS #	2367587-02-6
PubChem CID	155337814
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	5
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	4
Heavy Atom Count	28
Complexity	673
Defined Atom Stereocenter Count	3
SMILES	C1=CC(=NC=C1N)C(=O)NC2=NC(=O)N(C=C2)[C@H]3C([C@@H]([C@H](O3)CO)O)(F)F.Cl
InChi Key	WYFKQNWSWILBHM-JOCNCMMXSA-N
InChi Code	InChI=1S/C15H15F2N5O5.ClH/c16-15(17)11(24)9(6-23)27-13(15)22-4-3-10(21-14(22)26)20-12(25)8-2-1-7(18)5-19-8;/h1-5,9,11,13,23-24H,6,18H2,(H,20,21,25,26);1H/t9-,11-,13-;/m1./s1
Chemical Name	5-amino-N-[1-[(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-2-oxopyrimidin-4-yl]pyridine-2-carboxamide;hydrochloride
HS Tariff Code	2934.99.9001
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)

Biological Activity

Targets	IC50: 6.4 μM (H1N1) and 5.0 μM (H1N1)[1]
ln Vitro	In a dose-dependent manner, viral polymerase-IN-1 hydrochloride (Compound 2h; 0.1, 1, 10 μM; overnight) decreases the expression of viral NP protein and viral RNA copies [1]. In human HeLa cells, influenza A virus PR8's polymerase activity is efficiently inhibited by viral polymerase-IN-1 hydrochloride (11, 33, 100 μM; 72 hours) in a dose-dependent manner [1]. With a CC50 value of more than 100 μM, a SI value >217.4 in Calu-3 cells, and an EC50 value of 0.46 μM, viral polymerase-IN-1 hydrochloride effectively inhibits SARS-CoV-2 infection [1].
ln Vivo	In addition to lowering viral RNA levels in mice lungs, viral polymerase-IN-1 hydrochloride (Compound 2h; 5 mg/kg; intraperitoneally injected; once daily; for 5 days; began 4 hours before viral infection) also prevented infection-mediated pulmonary infiltrates [1].
Cell Assay	Western Blot Analysis[1] Cell Types: MDCK cells infected PR8 virus Tested Concentrations: 0.1, 1, 10 μM Incubation Duration: Overnight Experimental Results: decreased viral NP protein expression and viral RNA copies in a dose-dependent manner.
Animal Protocol	Animal/Disease Models: Sixweeks old BALB/c female mice intranasal infection with maPR8[1] Doses: 5 mg/kg Route of Administration: IP; one time/day for 5 days, beginning 4 h prior to virus infection Experimental Results: Alleviated lung damage or diminished viral RNA replication.
References	[1]. Evaluation of Antiviral Activity of Gemcitabine Derivatives against Influenza Virus and Severe Acute Respiratory Syndrome Coronavirus 2. ACS Infect Dis. 2023 Apr 14;9(4):1033-1045.

Solubility Data

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

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.3823 mL	11.9113 mL	23.8226 mL
	5 mM	0.4765 mL	2.3823 mL	4.7645 mL
	10 mM	0.2382 mL	1.1911 mL	2.3823 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.