title: Efficacy and safety of liposomal clarithromycin and its effect on Pseudomonas aeruginosa virulence factors.
abstract: We investigated the efficacy and safety of liposomal clarithromycin formulations with different surface charges against clinical isolates of Pseudomonas aeruginosa from the lungs of cystic fibrosis (CF) patients. The liposomal clarithromycin formulations were prepared by the dehydration-rehydration method, and their sizes were measured using the dynamic-light-scattering technique. Encapsulation efficiency was determined by microbiological assay, and the stabilities of the formulations in biological fluid were evaluated for a period of 48 h. The MICs and minimum bactericidal concentrations (MBCs) of free and liposomal formulations were determined with P. aeruginosa strains isolated from CF patients. Liposomal clarithromycin activity against biofilm-forming P. aeruginosa was compared to that of free antibiotic using the Calgary Biofilm Device (CBD). The effects of subinhibitory concentrations of free and liposomal clarithromycin on bacterial virulence factors and motility on agar were investigated on clinical isolates of P. aeruginosa. The cytotoxicities of the liposome preparations and free drug were evaluated on a pulmonary epithelial cell line (A549). The average diameter of the formulations was >222 nm, with encapsulation efficiencies ranging from 5.7% to 30.4%. The liposomes retained more than 70% of their drug content during the 48-h time period. The highly resistant strains of P. aeruginosa became susceptible to liposome-encapsulated clarithromycin (MIC, 256 mg/liter versus 8 mg/liter; P < 0.001). Liposomal clarithromycin reduced the bacterial growth within the biofilm by 3 to 4 log units (P < 0.001), significantly attenuated virulence factor production, and reduced bacterial twitching, swarming, and swimming motilities. The clarithromycin-entrapped liposomes were less cytotoxic than the free drug (P < 0.001). These data indicate that our novel formulations could be a useful strategy to enhance the efficacy of clarithromycin against resistant P. aeruginosa strains that commonly affect individuals with cystic fibrosis.
authors.affiliation: Novel Drug and Vaccine Delivery Systems Facility, Department of Chemistry and Biochemistry, Laurentian University, Sudbury, Ontario, Canada.
authors.forename: Mai
authors.initials: M
authors.lastname: Alhajlan
authors.forename: Moayad
authors.initials: M
authors.lastname: Alhariri
authors.forename: Abdelwahab
authors.initials: A
authors.lastname: Omri
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects / Q000254:growth & development; D045744:Cell Line, Tumor; D017291:Clarithromycin / Q000009:adverse effects* / Q000494:pharmacology* / Q000627:therapeutic use; D003550:Cystic Fibrosis / Q000382:microbiology; D004847:Epithelial Cells / Q000187:drug effects*; D006801:Humans; D008081:Liposomes / Q000009:adverse effects* / Q000494:pharmacology* / Q000627:therapeutic use; D008168:Lung / Q000166:cytology / Q000187:drug effects* / Q000382:microbiology; D008826:Microbial Sensitivity Tests / Q000706:statistics & numerical data; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000254:growth & development / Q000302:isolation & purification / Q000472:pathogenicity; D037521:Virulence Factors / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D008081:Liposomes; D037521:Virulence Factors; D017291:Clarithromycin
references.citation: Curr Opin Microbiol. 2008 Dec;11(6):560-6
references.pmid: 18935979
references.citation: Infect Immun. 1998 Jan;66(1):43-51
references.pmid: 9423837
references.citation: Drug Discov Today Dis Mech. 2007;4(2):63-72
references.pmid: 18560471
references.citation: J Antimicrob Chemother. 2002 May;49(5):867-70
references.pmid: 12003986
references.citation: J Antimicrob Chemother. 2007 Oct;60(4):760-9
references.pmid: 17673475
references.citation: FEMS Microbiol Lett. 2010 Aug 1;309(1):25-34
references.pmid: 20546309
references.citation: Int J Pharm. 2008 Jun 24;358(1-2):278-84
references.pmid: 18448284
references.citation: Int J Pharm. 2008 Jul 9;359(1-2):254-63
references.pmid: 18485632
references.citation: ISRN Pharm. 2012;2012:636743
references.pmid: 22500244
references.citation: Mol Microbiol. 1998 Oct;30(2):295-304
references.pmid: 9791175
references.citation: Respir Res. 2011 Nov 07;12:148
references.pmid: 22059807
references.citation: Chest. 2004 Feb;125(2 Suppl):62S-69S; quiz 69S
references.pmid: 14872002
references.citation: Int J Antimicrob Agents. 2011 Feb;37(2):140-4
references.pmid: 21130608
references.citation: J Bacteriol. 2001 Dec;183(24):7044-52
references.pmid: 11717261
references.citation: Int J Antimicrob Agents. 2008 Apr;31(4):329-36
references.pmid: 18280117
references.citation: J Clin Invest. 1993 Oct;92(4):1875-80
references.pmid: 8104958
references.citation: Can J Physiol Pharmacol. 2011 Jun;89(6):419-27
references.pmid: 21815782
references.citation: Med Mal Infect. 2006 Feb;36(2):78-91
references.pmid: 16427231
references.citation: Clin Biochem Rev. 2009 Nov;30(4):153-77
references.pmid: 20011209
references.citation: J Photochem Photobiol B. 2012 Feb 6;107:35-44
references.pmid: 22209036
references.citation: J Antimicrob Chemother. 2008 Apr;61(4):859-68
references.pmid: 18305202
references.citation: Arzneimittelforschung. 2008;58(10):485-92
references.pmid: 19025056
references.citation: Diagn Microbiol Infect Dis. 2009 Jan;63(1):81-6
references.pmid: 18990530
references.citation: Int Microbiol. 2010 Dec;13(4):207-12
references.pmid: 21404215
references.citation: Drug Deliv. 2004 Jan-Feb;11(1):33-9
references.pmid: 15168789
references.citation: Pulm Pharmacol Ther. 2008 Aug;21(4):595-9
references.pmid: 18234534
references.citation: Antimicrob Agents Chemother. 1993 Sep;37(9):1749-55
references.pmid: 8239580
references.citation: J Antimicrob Chemother. 1996 Sep;38(3):465-73
references.pmid: 8889721
references.citation: AAPS PharmSciTech. 2010 Mar;11(1):241-6
references.pmid: 20151337
references.citation: J Biol Chem. 1993 Apr 5;268(10):7503-8
references.pmid: 8463280
references.citation: J Antimicrob Chemother. 2010 Apr;65(4):684-93
references.pmid: 20159770
references.citation: Clin Microbiol Rev. 2011 Jan;24(1):29-70
references.pmid: 21233507
references.citation: Cancer Treat Rev. 2011 Dec;37(8):633-42
references.pmid: 21330062
references.citation: Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9427-31
references.pmid: 7568146
references.citation: Ann N Y Acad Sci. 2011 Dec;1241:33-47
references.pmid: 22191525
references.citation: Can J Microbiol. 2003 Oct;49(10):613-24
references.pmid: 14663495
references.citation: Int J Antimicrob Agents. 2006 Mar;27(3):247-52
references.pmid: 16472992
references.citation: Arthritis Rheum. 2011 Aug;63(8):2226-37
references.pmid: 21480191
references.citation: Cell. 1995 Feb 24;80(4):525-7
references.pmid: 7867059
references.citation: J Antimicrob Chemother. 2009 Aug;64(2):317-25
references.pmid: 19465435
references.citation: J Cyst Fibros. 2012 Jul;11(4):312-5
references.pmid: 22395041
references.citation: Biochim Biophys Acta. 1997 Oct 23;1329(2):345-56
references.pmid: 9371426
references.citation: Expert Opin Drug Deliv. 2010 Dec;7(12):1343-58
references.pmid: 21029028
references.citation: Infect Immun. 2000 Mar;68(3):1048-53
references.pmid: 10678906
references.citation: Int J Pharm. 2006 Jan 13;307(2):244-50
references.pmid: 16289986
references.citation: Antimicrob Agents Chemother. 2000 Oct;44(10):2869-72
references.pmid: 10991876
references.citation: Pharm Res. 1994 Aug;11(8):1127-31
references.pmid: 7971713
references.citation: Int J Pharm. 2008 May 1;355(1-2):293-8
references.pmid: 18164881
references.citation: J Clin Gastroenterol. 2011 Nov-Dec;45(10):841-3
references.pmid: 21989277
references.citation: J Antimicrob Chemother. 2007 Nov;60(5):1064-73
references.pmid: 17846106
references.citation: Respir Care. 2009 May;54(5):641-57
references.pmid: 19393108
references.citation: PLoS One. 2009 May 28;4(5):e5724
references.pmid: 19479000
references.citation: Sci Transl Med. 2011 Mar 16;3(74):74ra24
references.pmid: 21411740
references.citation: Antimicrob Agents Chemother. 1994 Mar;38(3):523-7
references.pmid: 8203849
references.citation: Int J Pharm. 2009 May 21;373(1-2):141-6
references.pmid: 19429299
references.citation: Microbes Infect. 2000 Jul;2(9):1051-60
references.pmid: 10967285
references.citation: Nat Med. 2012 Apr 05;18(4):509-19
references.pmid: 22481418
references.citation: J Drug Target. 1993;1(4):303-10
references.pmid: 8069572
references.citation: J Antimicrob Chemother. 1993 May;31(5):681-8
references.pmid: 8392997
references.citation: J Antimicrob Chemother. 2006 Jan;57(1):139-41
references.pmid: 16291867
references.citation: Eur J Clin Microbiol Infect Dis. 1992 May;11(5):469-71
references.pmid: 1330564
references.citation: J Antimicrob Chemother. 2004 Dec;54(6):1103-8
references.pmid: 15486079
references.citation: Trends Cell Biol. 2011 Sep;21(9):543-51
references.pmid: 21727007
references.citation: Antimicrob Agents Chemother. 2008 Oct;52(10):3648-63
references.pmid: 18644954
references.citation: J Cyst Fibros. 2011 Dec;10(6):387-400
references.pmid: 21775220
references.citation: J Med Chem. 2012 Feb 9;55(3):1242-51
references.pmid: 22214395
references.citation: J Pak Med Assoc. 2004 Aug;54(8):440-1
references.pmid: 15461217
references.citation: Can Respir J. 2003 Nov-Dec;10(8):449-51
references.pmid: 14679410
references.citation: Respir Med. 2003 Jul;97(7):844-50
references.pmid: 12854636
references.citation: Antimicrob Agents Chemother. 2007 Nov;51(11):4071-6
references.pmid: 17709469
references.citation: Acta Microbiol Immunol Hung. 2010 Sep;57(3):235-45
references.pmid: 20870595
references.citation: Anal Biochem. 2001 Mar;290(2):330-7
references.pmid: 11237336
references.citation: J Liposome Res. 2010 Mar;20(1):84-96
references.pmid: 19621992
references.citation: J Pharm Pharmacol. 2011 Aug;63(8):999-1007
references.pmid: 21718282
references.citation: BMC Med. 2011 Apr 04;9:32
references.pmid: 21463524
references.citation: Curr Opin Microbiol. 2012 Feb;15(1):71-7
references.pmid: 22137884
references.citation: Am J Respir Crit Care Med. 2005 Jun 1;171(11):1209-23
references.pmid: 15695491
references.citation: J Antimicrob Chemother. 2005 Feb;55(2):269-71
references.pmid: 15590716
references.citation: Antimicrob Agents Chemother. 2009 Apr;53(4):1552-60
references.pmid: 19188376
references.citation: J Infect Chemother. 2007 Dec;13(6):357-67
references.pmid: 18095083
references.citation: J Water Health. 2009;7 Suppl 1:S2-S18
references.pmid: 19717929
references.citation: J Clin Med Res. 2011 Oct;3(5):209-12
references.pmid: 22383907
references.citation: Clin Microbiol Rev. 2010 Jul;23(3):590-615
references.pmid: 20610825
references.citation: Phytopathology. 2012 Jun;102(6):575-87
references.pmid: 22568815
references.citation: Curr Allergy Asthma Rep. 2006 Mar;6(2):171-81
references.pmid: 16566868
references.citation: J Antimicrob Chemother. 2008 Dec;62(6):1291-7
references.pmid: 18931388
references.citation: Antimicrob Agents Chemother. 2006 Jun;50(6):2016-22
references.pmid: 16723560
references.citation: Front Microbiol. 2011 Aug 01;2:162
references.pmid: 21863142
references.citation: J Antimicrob Chemother. 2003 Aug;52(2):247-52
references.pmid: 12837733
references.citation: Antimicrob Agents Chemother. 2011 Apr;55(4):1588-93
references.pmid: 21220534