antiviral compound 34 is a potent and orally bioactive antiviral against influenza A and B subtypes with EC50 of 0.8 nM for H1N1 proliferation. antiviral compound 34 inhibits influenza virus proliferation by targeting influenza virus RNA-dependent RNA polymerase. antiviral compound 34 may be utilized in influenza virus research.

Physicochemical Properties

Molecular Formula	C29H33N3O2S
Molecular Weight	487.656225919724
Exact Mass	487.229
CAS #	945152-88-5
PubChem CID	17235233
Appearance	Typically exists as solid at room temperature
LogP	7.7
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	6
Heavy Atom Count	35
Complexity	730
Defined Atom Stereocenter Count	0
SMILES	S=C(NC1C=CC(=CC=1)NC(C1C=CC(=CC=1)C(C)(C)C)=O)NC(C1C=CC(=CC=1)C(C)(C)C)=O
InChi Key	GKGUZBBYDHKNIJ-UHFFFAOYSA-N
InChi Code	InChI=1S/C29H33N3O2S/c1-28(2,3)21-11-7-19(8-12-21)25(33)30-23-15-17-24(18-16-23)31-27(35)32-26(34)20-9-13-22(14-10-20)29(4,5)6/h7-18H,1-6H3,(H,30,33)(H2,31,32,34,35)
Chemical Name	4-tert-butyl-N-[4-[(4-tert-butylbenzoyl)carbamothioylamino]phenyl]benzamide
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

ln Vitro	Antiviral agent 34 (Compound 10m) has a high level of biosafety and has the ability to prevent the H1N1 virus from multiplying during a 24-hour period (0-470 nM) [1]
ln Vivo	In a mouse model of the deadly influenza virus, antiviral agent 34 (Compound 10m) (1 g/kg, given orally, three times a day for 14 days) demonstrated antiviral activity [1]. Route Dose (mg/kg) AUClast (ng·h/mL) AUCINF_obs (ng·h/mL) t1/2 (h) Tmax (h) Cmax (ng/mL) Clobs (L •h/kg) Vss_obs (mL/kg) MRTINF_obs (h) F (%) iv 3 21944 21920 3.69 0.08 41942 2.29 / / / po 10 5416 5395 3.26 4.0d 1184 / / / 7.38
Cell Assay	Cell Viability Assay[1] Cell Types: MDCK cells Tested Concentrations: 0-470 nM Incubation Duration: 24 h Experimental Results: demonstrated low toxicity to MDCK cells. Western Blot Analysis[1] Cell Types: MDCK cells Tested Concentrations: 2-125 nM Incubation Duration: 24 h Experimental Results: decreased the protein level of H1N1 nucleocapsid protein (NP) with low nanomolar values. Real Time qPCR[1] Cell Types: MDCK cells Tested Concentrations: 2-31.3 nM Incubation Duration: 24 h Experimental Results: decreased the mRNA level of H1N1 nucleocapsid protein (NP) with low nanomolar values.
Animal Protocol	Animal/Disease Models: balb/c (Bagg ALBino) mouse: [1] Doses: 1 g/kg/d continues for 14 days Route of Administration: po (oral gavage) Experimental Results: Tolerated at a dose of 1 g/kg with low acute toxicity. Provided ~ 50% protection in mice infected with lethal-dose H1N1. Animal/Disease Models: SD rats (pharmacokinetic/PK assay)[1] Doses: 3 mg/kg, 10 mg/kg Route of Administration: intravenous (iv)injection, oral administration (po) Experimental Results: Had preferable AUC value and low clearance rate and displayed superior oral bioavailability (F = 7.38%).
References	[1]. Liu X, et al. Rational design and optimization of acylthioureas as novel potent influenza virus non-nucleoside polymerase inhibitors. Eur J Med Chem. 2023 Nov 5;259:115678.

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	2.0506 mL	10.2530 mL	20.5061 mL
	5 mM	0.4101 mL	2.0506 mL	4.1012 mL
	10 mM	0.2051 mL	1.0253 mL	2.0506 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.