Leritrelvir (RAY1216) is an orally bioactive slow inhibitor of the major SARS-CoV-2 protease with Ki of 8.6 nM.

Physicochemical Properties

Molecular Formula	C31H44F3N5O6
Molecular Weight	639.71
Exact Mass	639.324
CAS #	2923310-64-7
PubChem CID	167713195
Appearance	White to off-white solid powder
LogP	3.6
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	10
Heavy Atom Count	45
Complexity	1170
Defined Atom Stereocenter Count	6
SMILES	C([C@@H]1[C@@]2([H])CCC[C@@]2([H])CN1C(=O)[C@H](C1CCCCC1)NC(=O)C(F)(F)F)(=O)N[C@H](C(=O)C(=O)NC1CCCC1)C[C@]1([H])CCNC1=O
InChi Key	ICGMMLTUQDVAST-HEZDJTGRSA-N
InChi Code	InChI=1S/C31H44F3N5O6/c32-31(33,34)30(45)38-23(17-7-2-1-3-8-17)29(44)39-16-19-9-6-12-21(19)24(39)27(42)37-22(15-18-13-14-35-26(18)41)25(40)28(43)36-20-10-4-5-11-20/h17-24H,1-16H2,(H,35,41)(H,36,43)(H,37,42)(H,38,45)/t18-,19-,21-,22-,23-,24-/m0/s1
Chemical Name	(3S,3aS,6aR)-2-[(2S)-2-cyclohexyl-2-[(2,2,2-trifluoroacetyl)amino]acetyl]-N-[(2S)-4-(cyclopentylamino)-3,4-dioxo-1-[(3S)-2-oxopyrrolidin-3-yl]butan-2-yl]-3,3a,4,5,6,6a-hexahydro-1H-cyclopenta[c]pyrrole-3-carboxamide
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	Ki: 8.6 nM (SARS-CoV-2 main protease)[1]
ln Vitro	The medication on-target residence period of leritrelvir (RAY1216) is 104 minutes [1]. Using an α-ketoamide warhead, leritrelvir is covalently bonded to the catalytic Cys145 [1]. Leritrelvir (0-1000 nM; 72 h) exhibits antiviral activity against SARS-CoV-2 mutant strains as well as the wild-type parent [1].
ln Vivo	Effectively, leritrelvir (RAY1216) (150–600 mg/kg/day; i.g., 5 days) prolongs the survival of mice infected with SARS-CoV-2 [1]. Compound pharmacokinetic characteristics across many animal taxa[1] Mixture Dose per species (mg/kg) Cmax (nM) AUC(0-last) (nM·h) Tmax (h) Cl (mL/min/kg) Vdss (L/kg) T1/2 (hour) oral F (%) 3.0 (IV) Mouse - - 7789 10 1.4 3.8 - 10 (PO) 1287 2.0 5698 - - 2.6 22 2.0 (IV) - 4505 12.5 1.1 2.2 - 10 (PO) 916 0.9 7429 - 4.3 33 Leritrelvir rat 1.0 (IV) - - 1157 22.5 1.0 0.9 - 5.0 (PO) 102 1.5 458 - - 14.9 8 cynomolgus macaque Cmax: the highest recorded dosage of the medication found in body fluids taken from clinical research participants Tmax: the amount of time needed to get the highest concentration, also known as the Cmax AUC. "Area Under the Curve" refers to the patient's overall drug exposure during a clinical research. Cl: complete clearing of plasma Vdss Constant state distribution volume T1/2: Half-time is the amount of time needed for half of the oral medication concentration to be removed. (F%) Bioavailability of oral SARS-CoV-2 infection model using a female transgenic human ACE2 C57BL/6 mice as the animal model[1]. The dosage is 150, 300, or 600 mg/kg per day. Administration: Five days of intragastric administration Result: At 600, 300, and 150 mg/kg, respectively, 100%, 43%, and 14% of protected mice were infected with SARS-CoV-2. considerably lower viral titres in the lungs when compared to the group that was just infected. decreased illness caused by viruses.
Cell Assay	Cell Viability Assay[1] Cell Types: Vero E6 cells inoculated with SARS-CoV-2 WT, Alpha, Beta, Delta, Omicron BA.1 and Omicron 247 BA.5 strains Tested Concentrations: 0-1000 nM Incubation Duration: 72 h Experimental Results: The half-maximal effective concentration (EC50) values against different SARS-CoV-2 variants were 95 nM (WT), 130 nM ( Alpha), 277 nM (Beta), 97 nM (Delta), 86 nM (Omicron BA.1) and 158 nM (Omicron BA.5), respectively.
Animal Protocol	Animal/Disease Models: Male CD-1 mouse, male SD rat and male cynomolgus macaque[1] Doses: 1-10 mg/kg Route of Administration: PO or IV (pharmacokinetic/PK Analysis) Experimental Results: demonstrated promising human pharmacokinetic/PKs profile.
References	[1]. Inhibition mechanism and antiviral activity of an α-ketoamide based SARS-CoV-2 main protease inhibitor. bioRxiv, 2023: 2023.03. 09.531862.

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	1.5632 mL	7.8160 mL	15.6321 mL
	5 mM	0.3126 mL	1.5632 mL	3.1264 mL
	10 mM	0.1563 mL	0.7816 mL	1.5632 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.