title: Drug repurposing screen reveals FDA-approved inhibitors of human HMG-CoA reductase and isoprenoid synthesis that block Cryptosporidium parvum growth.
abstract: Cryptosporidiosis, a diarrheal disease usually caused by Cryptosporidium parvum or Cryptosporidium hominis in humans, can result in fulminant diarrhea and death in AIDS patients and chronic infection and stunting in children. Nitazoxanide, the current standard of care, has limited efficacy in children and is no more effective than placebo in patients with advanced AIDS. Unfortunately, the lack of financial incentives and the technical difficulties associated with working with Cryptosporidium parasites have crippled efforts to develop effective treatments. In order to address these obstacles, we developed and validated (Z' score = 0.21 to 0.47) a cell-based high-throughput assay and screened a library of drug repurposing candidates (the NIH Clinical Collections), with the hopes of identifying safe, FDA-approved drugs to treat cryptosporidiosis. Our screen yielded 21 compounds with confirmed activity against C. parvum growth at concentrations of <10 μM, many of which had well-defined mechanisms of action, making them useful tools to study basic biology in addition to being potential therapeutics. Additional work, including structure-activity relationship studies, identified the human 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitor itavastatin as a potent inhibitor of C. parvum growth (50% inhibitory concentration [IC(50)] = 0.62 μM). Bioinformatic analysis of the Cryptosporidium genomes indicated that the parasites lack all known enzymes required for the synthesis of isoprenoid precursors. Additionally, itavastatin-induced growth inhibition of C. parvum was partially reversed by the addition of exogenous isopentenyl pyrophosphate, suggesting that itavastatin reduces Cryptosporidium growth via on-target inhibition of host HMG-CoA reductase and that the parasite is dependent on the host cell for synthesis of isoprenoid precursors.
authors: Department of Microbiology and Molecular Genetics, University of Vermont College of Medicine, Burlington, Vermont, USA.
authors: Kovi
authors: K
authors: Bessoff
authors: Adam
authors: A
authors: Sateriale
authors: K Kyungae
authors: KK
authors: Lee
authors: Christopher D
authors: CD
authors: Huston
mesh_terms: D002460:Cell Line; D003457:Cryptosporidiosis / Q000378:metabolism; D016785:Cryptosporidium parvum / Q000187:drug effects* / Q000378:metabolism*; D045782:Hemiterpenes / Q000494:pharmacology; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D020128:Inhibitory Concentration 50; D009943:Organophosphorus Compounds / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology; D013729:Terpenes / Q000494:pharmacology*
chemical_list: D045782:Hemiterpenes; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D009943:Organophosphorus Compounds; D011804:Quinolines; D013729:Terpenes; C004809:isopentenyl pyrophosphate; C108475:pitavastatin
references: J Antimicrob Chemother. 1996 Sep;38(3):399-408
references: 8889715
references: Acta Vet Hung. 1999;47(2):207-16
references: 10344081
references: Cell Microbiol. 2013 Jul;15(7):1182-97
references: 23311949
references: Health Aff (Millwood). 2006 Mar-Apr;25(2):420-8
references: 16522582
references: CMAJ. 2011 Nov 8;183(16):E1189-202
references: 21989464
references: Am J Trop Med Hyg. 1999 Nov;61(5):707-13
references: 10586898
references: Antimicrob Agents Chemother. 2005 Nov;49(11):4437-42
references: 16251280
references: Exp Parasitol. 2007 Aug;116(4):375-84
references: 17399705
references: Eur J Biochem. 2002 Jul;269(14):3339-54
references: 12135472
references: N Engl J Med. 1994 Nov 3;331(18):1173-80
references: 7935654
references: J Biomol Screen. 1999;4(2):67-73
references: 10838414
references: Chem Biol. 2008 Dec 22;15(12):1296-306
references: 19101474
references: Nat Rev Drug Discov. 2004 Aug;3(8):673-83
references: 15286734
references: Science. 1999 Sep 3;285(5433):1573-6
references: 10477522
references: Science. 2004 Apr 16;304(5669):441-5
references: 15044751
references: J Clin Gastroenterol. 1996 Jun;22(4):311-2
references: 8771430
references: Expert Opin Drug Saf. 2002 Sep;1(3):207-12
references: 12904136
references: J Exp Med. 2011 Jul 4;208(7):1547-59
references: 21690250
references: FEMS Microbiol Lett. 1999 Sep 15;178(2):227-33
references: 10499272
references: PLoS Pathog. 2011 Dec;7(12):e1002323
references: 22216001
references: Antimicrob Agents Chemother. 2001 Jun;45(6):1746-50
references: 11353621
references: Acta Paediatr Scand. 1987 May;76(3):474-7
references: 3604664
references: Semin Pediatr Infect Dis. 2004 Oct;15(4):229-36
references: 15494946
references: Antimicrob Agents Chemother. 2007 Jul;51(7):2654-5
references: 17502414
references: Cell Microbiol. 2011 Apr;13(4):569-86
references: 21105984
references: Atheroscler Suppl. 2011 Nov;12(3):271-6
references: 22152281
references: PLoS Negl Trop Dis. 2010 Aug 10;4(8):e794
references: 20706578
references: Cochrane Database Syst Rev. 2007 Jan 24;(1):CD004932
references: 17253532
references: PLoS Biol. 2011 Aug;9(8):e1001138
references: 21912516
references: J Antimicrob Chemother. 2000 Jul;46(1):57-60
references: 10882689
references: Int J Antimicrob Agents. 2009 Feb;33(2):185-6
references: 18996682
references: J Antimicrob Chemother. 2001 Jan;47(1):97-9
references: 11152438
references: Clin Microbiol Rev. 2002 Jul;15(3):390-400
references: 12097247
references: Clin Microbiol Rev. 2004 Jan;17(1):72-97
references: 14726456
references: Clin Chim Acta. 2007 Nov-Dec;386(1-2):25-30
references: 17719022
references: Biosci Biotechnol Biochem. 2002 Aug;66(8):1619-27
references: 12353619
references: Science. 2000 Mar 17;287(5460):1979-81
references: 10755953
references: Clin Infect Dis. 1998 Jan;26(1):146-50
references: 9455524
references: J Infect Dev Ctries. 2007 Dec 01;1(3):242-56
references: 19734601
references: N Engl J Med. 2002 May 30;346(22):1723-31
references: 12037153
references: Nucleic Acids Res. 2012 Jan;40(Database issue):D1100-7
references: 21948594
references: Cardiovasc Drugs Ther. 2002 May;16(3):251-7
references: 12374904
references: Microbiology (Reading). 2000 Feb;146 ( Pt 2):315-321
references: 10708370
references: Nucleic Acids Res. 1999 Jan 1;27(1):29-34
references: 9847135
references: Nucleic Acids Res. 1997 Sep 1;25(17):3389-402
references: 9254694
references: Antimicrob Agents Chemother. 1990 May;34(5):775-80
references: 2360817
references: Clin Cancer Res. 2012 Jul 1;18(13):3524-31
references: 22529099
references: J Eukaryot Microbiol. 1999 Sep-Oct;46(5):56S-57S
references: 10519247
references: Nucleic Acids Res. 2011 Jan;39(Database issue):D1035-41
references: 21059682
references: Antimicrob Agents Chemother. 1994 May;38(5):1144-8
references: 8067753
references: J Biol Chem. 2010 May 21;285(21):15916-22
references: 20231284
references: Nucleic Acids Res. 2010 Jan;38(Database issue):D415-9
references: 19914931