title: Inhibition of Mycoplasma pneumoniae growth by FDA-approved anticancer and antiviral nucleoside and nucleobase analogs.
abstract: BACKGROUND
  Mycoplasma pneumoniae (Mpn) is a human pathogen that causes acute and chronic respiratory diseases and has been linked to many extrapulmonary diseases. Due to the lack of cell wall, Mpn is resistant to antibiotics targeting cell wall synthesis such as penicillin. During the last 10 years macrolide-resistant Mpn strains have been frequently reported in Asian countries and have been spreading to Europe and the United States. Therefore, new antibiotics are needed. In this study, 30 FDA-approved anticancer or antiviral drugs were screened for inhibitory effects on Mpn growth and selected analogs were further characterized by inhibition of target enzymes and metabolism of radiolabeled substrates.
  
  
  RESULTS
  Sixteen drugs showed varying inhibitory effects and seven showed strong inhibition of Mpn growth. The anticancer drug 6-thioguanine had a MIC (minimum inhibitory concentration required to cause 90% of growth inhibition) value of 0.20 μg ml(-1), whereas trifluorothymidine, gemcitabine and dipyridamole had MIC values of approximately 2 μg ml(-1). In wild type Mpn culture the presence of 6-thioguanine and dipyridamole strongly inhibited the uptake and metabolism of hypoxanthine and guanine while gemcitabine inhibited the uptake and metabolism of all nucleobases and thymidine. Trifluorothymidine and 5-fluorodeoxyuridine, however, stimulated the uptake and incorporation of radiolabeled thymidine and this stimulation was due to induction of thymidine kinase activity. Furthermore, Mpn hypoxanthine guanine phosphoribosyl transferase (HPRT) was cloned, expressed, and characterized. The 6-thioguanine, but not other purine analogs, strongly inhibited HPRT, which may in part explain the observed growth inhibition. Trifluorothymidine and 5-fluorodeoxyuridine were shown to be good substrates and inhibitors for thymidine kinase from human and Mycoplasma sources.
  
  
  CONCLUSIONS
  We have shown that several anticancer and antiviral nucleoside and nucleobase analogs are potent inhibitors of Mpn growth and that the mechanism of inhibition are most likely due to inhibition of enzymes in the nucleotide biosynthesis pathway and nucleoside transporter. Our results suggest that enzymes in Mycoplasma nucleotide biosynthesis are potential targets for future design of antibiotics against Mycoplasma infection.
authors.affiliation: Department of Anatomy, Physiology, and Biochemistry, Swedish University of Agricultural Sciences, The Biomedical Centre, Uppsala, Sweden.
authors.forename: Ren
authors.initials: R
authors.lastname: Sun
authors.forename: Liya
authors.initials: L
authors.lastname: Wang
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006801:Humans; D008826:Microbial Sensitivity Tests; D009177:Mycoplasma pneumoniae / Q000187:drug effects* / Q000254:growth & development*; D009705:Nucleosides / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D009705:Nucleosides
references.citation: Gut. 2006 Oct;55(10):1423-31
references.pmid: 16543290
references.citation: Microbiol Mol Biol Rev. 1998 Dec;62(4):1094-156
references.pmid: 9841667
references.citation: Proc Natl Acad Sci U S A. 2004 Dec 28;101(52):17970-5
references.pmid: 15611477
references.citation: FEMS Microbiol Lett. 2005 Jun 15;247(2):193-8
references.pmid: 15927419
references.citation: J Bacteriol. 1977 Jun;130(3):1047-54
references.pmid: 324972
references.citation: Biochem Pharmacol. 2000 Jun 15;59(12):1583-8
references.pmid: 10799656
references.citation: J Med Chem. 2012 Nov 26;55(22):10010-21
references.pmid: 23043329
references.citation: Mol Microbiol. 2010 Sep;77(6):1502-11
references.pmid: 20860090
references.citation: Crit Rev Microbiol. 1997;23(4):269-354
references.pmid: 9439886
references.citation: Biochemistry. 1971 Sep 14;10(19):3598-607
references.pmid: 5146573
references.citation: ACS Chem Biol. 2012 Nov 16;7(11):1866-72
references.pmid: 22908966
references.citation: Pediatr Infect Dis J. 2012 Feb;31(2):208-9
references.pmid: 21941216
references.citation: Gene. 2007 Jul 15;396(2):215-25
references.pmid: 17467928
references.citation: Curr Top Med Chem. 2012;12(7):694-705
references.pmid: 22283813
references.citation: J Infect Chemother. 2010 Apr;16(2):78-86
references.pmid: 20094751
references.citation: Euro Surveill. 2012 Feb 02;17(5):
references.pmid: 22321134
references.citation: Biochim Biophys Acta. 2002 Jul 18;1587(2-3):206-14
references.pmid: 12084462
references.citation: Antimicrob Agents Chemother. 2007 Aug;51(8):2726-32
references.pmid: 17526755
references.citation: Mol Microbiol. 2001 Nov;42(4):1065-73
references.pmid: 11737647
references.citation: Proc Natl Acad Sci U S A. 2006 Jan 10;103(2):425-30
references.pmid: 16407165
references.citation: Hong Kong Med J. 2011 Oct;17(5):407-9
references.pmid: 21979480
references.citation: Antimicrob Agents Chemother. 2012 Feb;56(2):1108-9
references.pmid: 22106216
references.citation: Nucleosides Nucleotides Nucleic Acids. 2004 Oct;23(8-9):1499-502
references.pmid: 15571285
references.citation: Curr Med Chem. 2003 Jun;10(11):925-53
references.pmid: 12678681
references.citation: FEBS Lett. 1999 Jan 25;443(2):170-4
references.pmid: 9989599
references.citation: PLoS One. 2012;7(5):e37724
references.pmid: 22662200
references.citation: J Med Chem. 2012 Jan 26;55(2):852-70
references.pmid: 22243413
references.citation: Clin Infect Dis. 2010 Jul 15;51(2):189-94
references.pmid: 20540621
references.citation: Pediatr Infect Dis J. 2012 Apr;31(4):409-0
references.pmid: 22209916
references.citation: Bioorg Med Chem Lett. 2007 Nov 15;17(22):6239-44
references.pmid: 17890082
references.citation: Microbiol Immunol. 2001;45(8):617-20
references.pmid: 11592636
references.citation: J Bacteriol. 1979 Mar;137(3):1073-80
references.pmid: 220209
references.citation: J Bacteriol. 1978 Jun;134(3):706-12
references.pmid: 207675
references.citation: Antiviral Res. 2006 Sep;71(2-3):254-9
references.pmid: 16837073
references.citation: FEBS J. 2007 Apr;274(8):1983-90
references.pmid: 17355283
references.citation: Future Microbiol. 2011 Apr;6(4):423-31
references.pmid: 21526943
references.citation: Mol Microbiol. 2003 Nov;50(3):771-80
references.pmid: 14617140
references.citation: Inflamm Bowel Dis. 2012 Jan;18(1):63-73
references.pmid: 21381155
references.citation: Vet J. 2012 Oct;194(1):40-7
references.pmid: 22516918
references.citation: Biol Chem. 2006 Dec;387(12):1575-81
references.pmid: 17132103
references.citation: Int J Syst Bacteriol. 1996 Oct;46(4):885-90
references.pmid: 8863414
references.citation: J Infect Chemother. 2010 Jun;16(3):162-9
references.pmid: 20186455
references.citation: Clin Microbiol Rev. 2004 Oct;17(4):697-728, table of contents
references.pmid: 15489344
references.citation: Science. 1999 Dec 10;286(5447):2165-9
references.pmid: 10591650
references.citation: Ann Oncol. 1995 Nov;6(9):871-81
references.pmid: 8624289
references.citation: Mol Cell Biochem. 1994 Nov 9;140(1):1-22
references.pmid: 7877593
references.citation: Emerg Infect Dis. 2012 Feb;18(2):343-5
references.pmid: 22305177
references.citation: J Biol Chem. 2010 May 21;285(21):15916-22
references.pmid: 20231284
references.citation: Cold Spring Harb Perspect Med. 2012 Apr;2(4):a007161
references.pmid: 22474613
references.citation: FEBS J. 2010 Dec;277(23):4920-30
references.pmid: 21054786
references.citation: Science. 2009 Nov 27;326(5957):1263-8
references.pmid: 19965476
references.citation: Bioorg Med Chem. 2012 Jan 15;20(2):1076-89
references.pmid: 22178188