Identification of Novel Small-Molecule Inhibitors of Severe Acute Respiratory Syndrome-Associated Coronavirus by Chemical Genetics

Abstract The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) infected more than 8,000 people across 29 countries and caused more than 900 fatalities. Based on the concept of chemical genetics, we screened 50,240 structurally diverse small molecules from which we identified 104 compounds with anti-SARS-CoV activity. Of these 104 compounds, 2 target the SARS-CoV main protease (Mpro), 7 target helicase (Hel), and 18 target spike (S) protein-angiotensin-converting enzyme 2 (ACE2)-mediated viral entry. The EC50 of the majority of the 104 compounds determined by SARS-CoV plaque reduction assay were found to be at low micromolar range. Three selected compounds, MP576, HE602, and VE607, validated to be inhibitors of SARS-CoV Mpro, Hel, and viral entry, respectively, exhibited potent antiviral activity (EC50 < 10 μM) and comparable inhibitory activities in target-specific in vitro assays.

[1]  G. Gao,et al.  The crystal structures of severe acute respiratory syndrome virus main protease and its complex with an inhibitor , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[2]  T. Takeuchi,et al.  [Entamoeba histolytica]. , 1999, Nihon rinsho. Japanese journal of clinical medicine.

[3]  Obi L. Griffith,et al.  The Genome Sequence of the SARS-Associated Coronavirus , 2003, Science.

[4]  John L. Sullivan,et al.  Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus , 2003, Nature.

[5]  B. Stockwell.  Chemical genetics: ligand-based discovery of gene function , 2000, Nature Reviews Genetics.

[6]  J. Keldenich,et al.  New helicase-primase inhibitors as drug candidates for the treatment of herpes simplex disease , 2002, Nature Medicine.

[7]  M. Leippe,et al.  Recombinant expression and purification of an enzymatically active cysteine proteinase of the protozoan parasite Entamoeba histolytica. , 2002, Protein expression and purification.

[8]  Stuart L. Schreiber,et al.  Dissecting glucose signalling with diversity-oriented synthesis and small-molecule microarrays , 2002, Nature.

[9]  J. A. Comer,et al.  A novel coronavirus associated with severe acute respiratory syndrome. , 2003, The New England journal of medicine.

[10]  Rolf Hilgenfeld,et al.  Coronavirus Main Proteinase (3CLpro) Structure: Basis for Design of Anti-SARS Drugs , 2003, Science.

[11]  Y. Guan,et al.  Coronavirus as a possible cause of severe acute respiratory syndrome , 2003, The Lancet.

[12]  F. Javier Luque,et al.  Polarization effects in generalized molecular interaction potential: New Hamiltonian for reactivity studies and mixed QM/MM calculations , 1998 .

[13]  M. Cordingley,et al.  Herpes simplex virus helicase-primase inhibitors are active in animal models of human disease , 2002, Nature Medicine.

[14]  J. Dowling,et al.  Small molecule developmental screens reveal the logic and timing of vertebrate development. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[15]  David S. Goodsell,et al.  Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function , 1998 .

[16]  Alexander E Gorbalenya,et al.  Mechanisms and enzymes involved in SARS coronavirus genome expression. , 2003, The Journal of general virology.

[17]  M. Ng,et al.  Proliferative growth of SARS coronavirus in Vero E6 cells. , 2003, The Journal of general virology.

[18]  Christian Drosten,et al.  Identification of a novel coronavirus in patients with severe acute respiratory syndrome. , 2003, The New England journal of medicine.

[19]  S. Schreiber.  Chemical genetics resulting from a passion for synthetic organic chemistry. , 1998, Bioorganic & medicinal chemistry.

[20]  Julian A. Tanner,et al.  The Severe Acute Respiratory Syndrome (SARS) Coronavirus NTPase/Helicase Belongs to a Distinct Class of 5′ to 3′ Viral Helicases , 2003, Journal of Biological Chemistry.

[21]  Y. Guan,et al.  The Complete Genome Sequence of Severe Acute Respiratory Syndrome Coronavirus Strain HKU-39849 (HK-39) , 2003, Experimental biology and medicine.

[22]  M. Ng,et al.  Early events of SARS coronavirus infection in vero cells , 2003, Journal of medical virology.

[23]  J. Jenkins,et al.  A small-molecule inhibitor of the ribonucleolytic activity of human angiogenin that possesses antitumor activity , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Christian Drosten,et al.  Characterization of a Novel Coronavirus Associated with Severe Acute Respiratory Syndrome , 2003, Science.

[25]  T. Mitchison,et al.  Towards a pharmacological genetics. , 1994, Chemistry & biology.

[26]  Ralph S. Baric,et al.  Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[27]  Scott D. Kahn,et al.  Assessment of putative protein targets derived from the SARS genome , 2003, FEBS Letters.