Rifapentine"‘Proliposomes for Inhalation: In Vitro and In Vivo Toxicity
Keywords:A549 cell line, drug susceptibility testing on Mycobacteria growth indicator tube, pulmonary tuberculosis, rifapentine, toxicity
AbstractBackground: Oral therapy for pulmonary tuberculosis (TB) treatment suffers from the limitation of hepatic metabolism leading insufficient concentration of antitubercular (anti"‘TB) drugs in alveolar macrophage which harbors Mycobacterium tuberculosis (MTB). Targeted aerosol delivery of antituberculous drug to lung is efficient for treating local lung TB infection. Objective: The present study was aimed to evaluate rifapentine (RPT) loaded proliposomal dry powder for inhalation (RLDPI) for anti"‘TBactivity and cytotoxicity in vitro. In vivo toxicity study was also undertaken in Wistar rats to determine safe concentration of RLDPI for administration. Materials and Methods: Anti"‘TB activity of developed RLDPI was assessed using drug susceptibility testing (DST) on Mycobacteria growth indicator tube (MGIT) method. in vitro cytotoxicity was performed in A549 cell lines and IC50 values were used to compare the cytotoxicity of formulation with pure RPT. in vivo repeated dose toxicity study was undertaken using Wistar rats at three different doses for 28"‘days by intratracheal insufflations method. Results: The results of DST study revealed sensitivity of tubercle bacteria to RLDPI at concentration equivalent to 10 Î¼g/mL of RPT. This study confirmed anti"‘TB potential of RPT in spray"‘dried RLDPI, though the spray drying method is reported to reduce activity of drugs. Cytotoxicity study in A549 cells demonstrated that RPT when encapsulated in liposomes as RLDPI was safe to cells as compared to pure RPT. In vivo toxicity study revealed that RPT in the form of RLDPI was safe at 1 and 5 mg/kg dose. However, mortality was seen at higher dose (10 mg/kg), possibly because of liver and kidney damage. Conclusion: Thus, these studies demonstrated safety of RLDPI for the treatment of pulmonary TB.
World Health Organisation. Tuberculosis fact sheet. Available from: http://www.whi.int/inffs/en/fact104.html [Last accessed on 2014 Apr 28].
Pandey R, Khuller GK. Antitubercular inhaled therapy: Opportunities, progress and challenges. J Antimicrob Chemotherapy 2005;55:430"‘5.
Gelperina S, Kisich K, Iseman MD, Heifets L. The potential advantages of nanoparticle drug delivery systems in chemotherapy of tuberculosis. Am J Respir Crit Care Med 2005;172:1487"‘90.
Sharma R, Saxena D, Dwivedi AK, Misra A. Inhalable microparticles containing drug combinations to target alveolar macrophages for treatment of pulmonary tuberculosis. Pharm Res 2001;18:1405"‘10.
Suarez S, O'Hara P, Kazantseva M, Newcomer CE, Hopfer R, McMurray DN, et al. Respirable PLGA microspheres containing rifampicin for the treatment of tuberculosis: Screening in an infectious disease model. Pharm Res 2001;18:1315"‘9.
Sethuraman VV, Hickey AJ. Powder properties and their influence on dry powder inhaler delivery of an anti"‘tubercular drug. AAPS Pharm Sci Tech 2002;3:E28.
Muttil P, Kaur J, Kumar K, Yadav AB, Sharma R, Misra A. Inhalable microparticles containing large payload of anti"‘tuberculosis drugs. Eur J Pharm Sci 2007;32:140"‘50.
Misra A, Hickey AJ, Rossi C, Borchard G, Terada H, Makino K, et al. Inhaled drug therapy for treatment of tuberculosis. Tuberculosis (Edinb) 2010;91:71"‘81.
Weibel ER. Review: Morphometry of the human lung. Berlin: Springer Verlag: Academic Press; 1963. p. 3.
Kelly C, Jefferies C, Cryan SA. Targeted liposomal drug delivery to monocytes and macrophages. J Drug Deliv 2011;2011:727241.
Vyas SP, Kannan ME, Jain S, Mishra V, Singh P. Design of liposomal aerosols for improved delivery of rifampicin to alveolar macrophages. Int J Pharm 2004;269:37"‘49.
Wiethoff CM, Smith JG, Koe GS, Middaugh CR. The potential role of proteoglycans in cationic lipid"‘mediated gene delivery. Studies of the interaction of cationic lipid"‘DNA complexes with model glycosaminoglycans. J Biol Chem 2001;276:32806"‘13.
European Medicines Agency, Orphan designation. EU/3/06/387.
Available from: http://clinicaltrials.gov/ct2/show/ NCT01315678?term=ARIKACE and rank=5 [Last accessed on 2014 Feb 28].
Available from: http://www.aradigm.com/products_3100. html [Last accessed on 2014 Feb 28].
Jarvis B, Lamb HM. Rifapentine. Drugs 1998;56:607"‘16.
Drugs @ FDA. Available from: http://www.accessdata.fda.gov/ drugsatfda_docs/label/1998/21024lbl.pdf [Last accessed on 2014 Jan 02].
Patil"‘Gadhe A, Pokharkar V. Single step spray drying method to develop proliposomes for inhalation: A systematic study based on quality by design approach. Pulm Pharmacol Ther 2014;27:197"‘207.
Lin SY, Desmond E, Bonato D, Gross W, Siddiqi S. Multicenter evaluation of Bactec MGIT 960 system for second"‘line drug susceptibility testing of Mycobacterium tuberculosis complex. J Clin Microbiol 2009;47:3630"‘4.
Rüsch"‘Gerdes S, Pfyffer GE, Casal M, Chadwick M, Siddiqi S. Multicenter laboratory valuation of the BACTEC MGIT 960 technique for testing susceptibilities of Mycobacterium tuberculosis to classical second"‘line drugs and newer antimicrobials. J Clin Microbiol 2006;44:688"‘92.
Patlolla RR, Chougule M, Patel AR, Jackson T, Tata PN, Singh M. Formulation, characterization and pulmonary deposition of nebulized celecoxib encapsulated nanostructured lipid carriers. J Control Release 2010;144:233"‘41.
Rojanarat W, Nakpheng T, Thawithong E, Yanyium N, Srichana T. Levofloxacin"‘proliposomes: Opportunities for use in lung tuberculosis. Pharmaceutics 2012;4:385"‘412.
Brain JD, Knudson DE, Sorokin SP, Davis MA. Pulmonary distribution of particles given by intratracheal instillation or by aerosol inhalation. Environ Res 1976;11:13"‘33.
Sayes CM, Reed KL, Warheit DB. Assessing toxicity of fine and nanoparticles: Comparing in vitro measurements to in vivo pulmonary toxicity profiles. Toxicol Sci 2007;97:163"‘80.
Sanna V, Kirschvink N, Gustin P, Gavini E, Roland I, Delattre L, et al. Preparation and in vivo toxicity study of solid lipid microparticles as carrier for pulmonary administration. AAPS Pharm Sci Tech 2004;5:e27.
Warheit DB, Carakostas MC, Hartsky MA, Hansen JF. Development of a short"‘term inhalation bioassay to assess pulmonary toxicity of inhaled particles: Comparisons of pulmonary responses to carbonyl iron and silica. Toxicol Appl Pharmacol 1991;107:350"‘68.
Available form: http://tbonline.info/posts/2011/11/29/ drug"‘susceptibility"‘testing"‘mgit"‘system/[Last accessed on 2015 Feb 28].
Venkataraman P, Paramasivan CN, Prabhakar R. In vitro activity of rifampicin, rifapentine and rifabutin against south Indian isolates of mycobacterium tuberculosis. Indian J Tub 1993;40:17"‘20.
Drent M, Cobben NA, Henderson RF, Wouters EF, van Dieijen"‘Visser M. Usefulness of lactate dehydrogenase and its isoenzymes as indicators of lung damage or inflammation. Eur Respir J 1996;9:1736"‘42.