PF-00835231 (PF-835231) is a potent 3CLpro [main protease (Mpro)] inhibitor developed by Pfizer for COVID-19 treatment. It is the active component of the phosphate prodrug PF-07304814. 3CL protease is a virally encoded protein essentialfor a broad spectrum of coronaviruses with no close human analogs. PF-00835231 exhibited potent in vitro antiviral activity against SARS-CoV-2 as a single agent. PF-00835231 has additive/synergistic activity in combination with remdesivir.

Physicochemical Properties

Molecular Formula	C24H32N4O6
Molecular Weight	472.534086227417
Exact Mass	472.232
Elemental Analysis	C, 61.00; H, 6.83; N, 11.86; O, 20.31
CAS #	870153-29-0
Related CAS #	870153-29-0 70153-89-2 (racemic);870153-90-5; 870153-91-6;
PubChem CID	11561899
Appearance	White to off-white solid powder
LogP	1.6
Hydrogen Bond Donor Count	5
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	11
Heavy Atom Count	34
Complexity	760
Defined Atom Stereocenter Count	3
SMILES	CC(C)C[C@@H](C(=O)N[C@@H](C[C@@H]1CCNC1=O)C(=O)CO)NC(=O)C2=CC3=C(N2)C=CC=C3OC
InChi Key	QDIMHKWNHMVDJB-WBAXXEDZSA-N
InChi Code	InChI=1S/C24H32N4O6/c1-13(2)9-18(23(32)27-17(20(30)12-29)10-14-7-8-25-22(14)31)28-24(33)19-11-15-16(26-19)5-4-6-21(15)34-3/h4-6,11,13-14,17-18,26,29H,7-10,12H2,1-3H3,(H,25,31)(H,27,32)(H,28,33)/t14-,17-,18-/m0/s1
Chemical Name	N-((S)-1-(((S)-4-hydroxy-3-oxo-1-((S)-2-oxopyrrolidin-3-yl)butan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)-4-methoxy-1H-indole-2-carboxamide
Synonyms	PF 00835231; PF-00835231; PF00835231; PF 835231; PF-835231; PF835231;
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	3CLpro
References	[1]. Discovery of Ketone-Based Covalent Inhibitors of Coronavirus 3CL Proteases for the Potential Therapeutic Treatment of COVID-19. J Med Chem. 2020 Nov 12;63(21):12725-12747. [2]. Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19. Nat Commun. 2021 Oct 18;12(1):6055.
Additional Infomation	PF-00835231 is a primary alcohol resulting from the cleavage of the phosphate group of the prodrug PF-07304814. It is an inhibitor of SARS-CoV-1 and -2 main protease (3CLpro) and exhibits potent in vitro antiviral activity. It has a role as an anticoronaviral agent, an EC 3.4.22.69 (SARS coronavirus main proteinase) inhibitor and a drug metabolite. It is an aromatic ether, a L-leucine derivative, an indolecarboxamide, a member of pyrrolidin-2-ones, a secondary carboxamide and a primary alcohol.

Solubility Data

Solubility (In Vitro)

DMSO : ~250 mg/mL (~529.07 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.40 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 20.8 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. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.40 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. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (4.40 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1163 mL	10.5813 mL	21.1627 mL
	5 mM	0.4233 mL	2.1163 mL	4.2325 mL
	10 mM	0.2116 mL	1.0581 mL	2.1163 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.