{"paper_id":"43f3bbd5-0dcf-4cf2-80a2-ce0ef70fb8e7","body_text":"Abstract\nLeuprolide acetate (LPA), a GnRH analogue, is drug of choice for treatment of uterine fibroids and endometriosis. The current marketed formulations of LPA show severe systemic side effects. This project aims to formulate LPA loaded liposomes to be administered by vaginal route for uterine targeting. Liposomes were prepared by thin film hydration method using 1:1 M ratio of DSPC: Cholesterol and characterized for vesicle size, zeta potential, entrapment efficiency, and loading. Radiolabeling of LPA was performed by direct labeling with reduced technetium-99m. Binding affinity of 99mTc-labeled complexes was assessed by diethylenetriaminepentaacetic acid (DTPA) challenge test. Biodistribution study was done in New Zealand white female rabbits by administering the formulation via vaginal route. Spherical and discrete vesicles of size 189 nm were seen in TEM results with entrapment efficiency and loading of 74.36% and 9.29%w/w, respectively. Liposomes were able to sustain the drug release for 5 days. 99mTc-labeled complexes showed high labeling efficiency and stability both in saline and serum. DTPA challenge test confirmed low transchelation of 99mTc-labeled complexes. Biodistribution study by gamma scintigraphy revealed the preferential uptake of the formulation by uterus when administered vaginally. Compared to plain drug, liposomes concentrated and were retained within the uterus for a longer period of time. Uterine targeting of liposomal LPA indicates its potential to overcome the limitations of presently available formulations. Hence, this seems to be a promising approach for targeting the drugs, whose site of action is uterus.\nSimilar content being viewed by others\nReferences\nWhitfield CR, editor. Dewhurst’s textbook of obstetrics and gynaecology for post-gruduates. 4th ed. London: Blackwell Scientific Publications; 1986. p. 726–33.\nSimms-Stewart D, Fletcher H. Counselling patients with uterine fibroids: a review of the management and complications. Obstet Gynecol Int. 2012.\nRobboy SJ, Anderson MC, Russell P. Female reproductive tract pathology [M]. Translator: Hui YZ, et al. Beijing: Peking University Medical Press; 2005. pp 466–473. [in Chinese].\nSomigliana E, Vigano P, Parazzini F, Stoppelli S, Giambattista E, Vercellini P. Association between endometriosis and cancer: a comprehensive review and critical analysis of clinical and epidemiological evidence. Gynecol Oncol. 2006;101(2):331–41.\nBulun SE. Endometriosis. N Engl J Med. 2009;360(3):268–79.\nFlake GP, Andersen J, Dixon D. Etiology and pathogenesis of uterine leiomyomas: a review. Environ Health Perspect. 2003;111:1037–54.\nCook JD, Walker CL. Treatment strategies for uterine leiomyoma: the role of hormonal modulation. Semin Reprod Med. 2004;22:105–11.\nEnglund K, Blanck A, Gustavsson I, Lundkvist U, Sjoblom P, Norgren A, et al. Sex steroid receptors in human myometrium and fibroids: changes during the menstrual cycle and gonadotropin-releasing hormone treatment. J Clin Endocrinol Metab. 1998;83:4092–6.\nNisolle M, Gillerot S, Casanas-Roux F, Squifflet J, Berliere M, Donnez J. Immunohistochemical study of the proliferation index, oestrogen receptors and progesterone receptors A and B in leiomyomata and normal myometrium during the menstrual cycle and under gonadotrophin-releasing hormone agonist therapy. Hum Reprod. 1999;14:2844–50.\nAubuchon M, Pinto AB, Williams DB. Treatment of uterine fibroids. Obstet Gynaecol Reprod Med. 2002;9(6):231–7.\nLefebvre G, Vilos G, Allaire C, Jeffrey J, Arneja J, Birch C, et al. The management of uterine leiomyomas. J Obstet Gynaecol Can. 2003;25(5):396–418.\nGolan A. GnRH analogues in the treatment of uterine fibroids. Hum Reprod. 1996;11(suppl 3):33–41.\nLim SS, Sockalingam JK, Tan PC. Goserelin vs leuprolide before hysterectomy for uterine fibroids. Int J Gynecol Obstet. 2008;101:178–83.\nWilson AC, Meethal SV, Bowen RL, Atwood CS. Leuprolide acetate: a drug of diverse clinical applications. Exp Opin Investig Drugs. 2007;16(11):1–13.\nFicicioglu C, Kutlu T, Tasdemir S, Altuntas L. The effects of different gonadotropin releasing hormone analogues in IVF cycles. Middle East Fertil Soc J. 2005;10(3):212–9.\nMagon N. Gonadotropin releasing hormone agonists: expanding vistas. Indian J Endocrinol Metab. 2011;15(4):261.\nVega JA, Ochoa PS, Holder P. “Chapter 1: introduction to parenteral preparations” in Concepts in sterile preparations and aseptic technique. Burlington: Jones and Barlett Learning; 2015. p. 1–15.\nFulcher EM, Fulcher RM, Soto CD. “Chapter 14: parenteral routes” in Pharmacology—principles and applications, Third Edition. Saunders, Elsevier; 2012. pp. 218–229.\nMiles RA, Paulson RJ, Lobo RA, Press MF, Dahmoush L, Sauer MV. Pharmacokinetics and endometrial tissue levels of progesterone after administration by intramuscular and vaginal routes: a comparative study. Fertil Steril. 1994;62:485–90.\nPatel A, Tyagi A, Sharma RK, Thakkar H. A Gamma Scintigraphy study to investigate uterine targeting efficiency of raloxifene loaded liposomes administered intravaginally in New Zealand white female rabbits. Drug Deliv. 2016:1–36. https://doi.org/10.1080/10717544.2016.1177137.\nZiegler D, Cicinelli E. Transvaginal progesterone: evidence for a new functional ‘portal system’ flowing from the vagina to the uterus. Hum Reprod Update. 1999;5:365–72.\nNeves JD, Amaral MH, Bahia MF. Vaginal drug delivery ystems. In: Gad SC, editor. Pharmaceutical manufacturing handbook: production and processes: Hoboken: A John Wiley & Sons, Inc., Publications; 2008. pp. 809–878.\nPinheiro M, Lúcio M, Lima JL, Reis S. Liposomes as drug delivery systems for the treatment of TB. Nanomedicine. 2011;6(8):1413–28.\nAllison SD. Liposomal drug delivery. J Infus Nurs. 2007;30(2):89–95.\nDing H, Wu F. Image guided biodistribution and pharmacokinetic studies of Theranostics. Theranostics. 2012;2(11):1040–53.\nRandip K, Jili C, Dawoodji A, et al. Preparation, characterisation and entrapment of a non-glycosidic Threitol ceramide into liposomes for presentation to invariant natural killer T cells. J Pharm Sci. 2011;100:2724–33.\nSabeti B, Noordine MIB, Javar HA, Davoudi ET, Kadivar A. Characterization of diclofenac liposomes formulated with palm oil fractions. Trop J Pharm Res. 2014;13(2):185–90.\nRichardson VJ, Jeyasingh K, Jewkes RF. Properties of [99mTc] technetium- labeled liposomes in normal and tumour-bearing rats. Biochem Soc Trans. 1977;5(1):290–29.\nArulsudar N, Subramanian N, Mishra P, Chuttani K, Sharma RK, Murthy RSR. Preparation, characterization, and biodistribution study of Technetium99m labeled leuprolide acetate-loaded liposomes in Ehrlich ascites tumor-bearing mice. AAPS PharmSci. 2004;6(1):1–12.\nReddy LK, Sharma RK, Chuttani K, Mishra AK, Murthy RSR. Etoposide incorporated Tripalmitin nanoparticles with different surface charge: formation, characterization, radiolabeling and biodistribution studies. AAPS J. 2004;6(3):1–10.\nSingh T, Kumar N, Soni S, Rawat H, Mittal G, Singh AK, et al. A new method for radiolabeling of human immunoglobulin-G and its biological evaluation. J Pharm Bioallied Sci. 2012;4(4):286.\nShaw SR, Nihal M, Ahmad N. Dose translation from animal to human studies revisited. FASEB J. 2007;22:659–61.\nVerma DD, McElwee KJ, Freyschmidt- Paul P, Fahr A. Treatment of alopecia areata in the BEBR model using cyclosporine a lipid vesicles. Eur J Dermatol. 2004;14:332–8.\nCevc G, Blume G. Transdermal drug carriers: basic properties, optimization and transfer efficiency in the case of epicutaneously applied peptides. J Control Release. 1995;36:3–16.\nHood RR, Kendall EL, Junqueira M, Vreeland WN, Quezado Z, Finkel JC, et al. Microfluidic-enabled liposomes elucidate size-dependent transdermal transport. PLoS One. 2014;9(3):e92978.\nMurakami A, Fukada K, Yamano Y, Gohtani S. Effects of sugars on the D phase emulsification of triglyceride using polyoxyethylene sorbitan fatty acid ester. J Oleo Sci. 2005;54(12):633–9.\nCrooke ST. Antisense drug technology: principles, strategies and applications. Boca Raton: Taylor and Francis Group, CRC Press; 2008. p. 253.\nMohammed AR, Weston N, Fitzgerald CM, Perrie Y. Liposome formulation of poorly water soluble drugs: optimisation of drug loading and ESEM analysis of stability. Int J Pharm. 2004;28:23–34.\nKirby C, Gregoriadis G. Effect of the cholesterol content of small unilamellar liposomes on their stability in vivo and in vitro. Biochem J. 1980;186:591–8.\nBalny C, Masson P, Heremans K. Frontiers in high pressure biochemistry and biophysics. Amsterdam: Elsevier; 2002.\nDan. Lipid tail chain asymmetry and the strength of membrane-induced interactions between membrane proteins. Biochim Biophys Acta. 2007;1768:2393–9.\nKirby C, Gregoriadis G. Dehydration-rehydration vesicles: a simple method for high yield drug entrapment in liposomes. Biotechnology. 1984;2(11):979–84.\nSenior J, Crawley JC, Gregoriadis G. Tissue distribution of liposomes exhibiting long half-lives in the circulation after intravenous injection. Biochim Biophys Acta Gen Subj. 1985;839(1):1–8.\nCharrois GJ, Allen TM. Drug release rate influences the pharmacokinetics, biodistribution, therapeutic activity, and toxicity of pegylated liposomal doxorubicin formulations in murine breast cancer. Biochim Biophys Acta Biomembr. 2004;1663(1):167–77.\nBanerjee S, Pillai MRA, Ramamoorthy N. Evolution of Tc-99m in diagnostic radiopharmaceuticals. Semin Nucl Med. 2001;31:260–77.\nMajumdar D, Saha CN, Bhattacharya S. 99mTechnetium radiolebeling and biodistribution studies of some peptide based ligands. World J Med Sci. 2011;6(3):105–10.\nMoustapha ME, Shweeta HA, Motaleb MA. Technetium-labeled danofloxacin complex as a model for infection imaging. Arab J Chem. 2014:1–7. https://doi.org/10.1016/j.arabjc.2014.10.017.\nCastronovo FP, Deluca SA, Potsaid MS. 99m Tc (Sn) phosphocreatine: labeling characteristics and biological distribution. Int J Appl Radiat Isot. 1976;27(2):93–6.\nBulletti C, Ziegler DD, Giacomucci E, Polli V, Rossi S, Alfieri S, et al. Vaginal drug delivery: the first uterine pass effecta. Ann N Y Acad Sci. 1997;828(1):285–90.\nCicinelli E, Rubini G, et al. Absorption and preferential vagina-to-uterus distribution after vaginal administration of 99mTc-pertechnetate in postmenopausal women. Fertil Steril. 2001;76(6):1108–12.\nRefeurzo JS, Alexander JF, Leonard F, et al. Liposomes: a nanoscale drug carrying system to prevent indomethacin passage to the fetus in a pregnant mouse model. Am J Obstet Gynecol. 2015;212:508–10.\nAuthor information\nAuthors and Affiliations\nCorresponding author\nEthics declarations\nEthics approval\nAll institutional and national guidelines for the care and use of laboratory animals were followed.\nConflict of interest\nThe authors declare that they have no conflicts of interest.\nRights and permissions\nAbout this article\nCite this article\nPatel, A., Tyagi, A., Sharma, R.K. et al. Formulation of 99mTechnetium-labeled leuprolide loaded liposomes and its biodistribution study in New Zealand white female rabbits for assessment of its uterine targeting efficiency. Drug Deliv. and Transl. Res. 8, 43–53 (2018). https://doi.org/10.1007/s13346-017-0432-1\nPublished:\nIssue date:\nDOI: https://doi.org/10.1007/s13346-017-0432-1","source_license":"CC0","license_restricted":false}