title: Investigations on feasibility of in situ development of amphotericin B liposomes for industrial applications.
abstract: Amphotericin B (AmB) liposome formulations are very successful in the treatment of fungal infections and leishmaniasis. But higher cost limits its widespread use among people in developing countries. Therefore, we have developed a modified ethanol-injection method for the preparation of AmB liposomes. Two liposomal formulations were developed with dimyristoyl phosphatidylcholine [F-1a] and soya phosphatidylcholine [F-2a], along with egg phosphatidyl glycerol and cholesterol. AmB was dissolved in acidified dimethyl acetamide and mixed with ethanolic lipid solution and rapidly injected in 5% dextrose to prepare liposomes. Liposomes were characterized on the basis of size (~100 nm), zeta (-43.3 ± 2.8 mV) and percent entrapment efficiency (>95%). The in vitro release study showed an insignificant difference (P ≥ 0.05) for 24-hour release between marketed AmB liposomes (AmBisome) and F-1a and F-2a. Proliposome concentrate, used for the preparation of in situ liposomes, was physically stable for more than 3 months at experimental conditions. Similarly, AmB showed no sign of degradation in reconstituted liposomes stored at 2-8°C for more than 3 months. IC(50) value of Ambisome (0.18 µg/mL) was comparatively similar to F-1a (0.17 µg/mL) and F-2a (0.16 µg/mL) against intramacrophagic amastigotes. Under experimental conditions, a novel modified method for AmB liposomes is a great success and generates interest for development as a platform technology for many therapeutic drug products.
authors.affiliation: Pharmaceutics Division, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Deepak
authors.initials: D
authors.lastname: Singodia
authors.forename: Ashwni
authors.initials: A
authors.lastname: Verma
authors.forename: Prashant
authors.initials: P
authors.lastname: Khare
authors.forename: Anuradha
authors.initials: A
authors.lastname: Dube
authors.forename: Kalyan
authors.initials: K
authors.lastname: Mitra
authors.forename: Prabhat Ranjan
authors.initials: PR
authors.lastname: Mishra
mesh_terms: D000666:Amphotericin B / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004355:Drug Stability; D005374:Filtration; D006461:Hemolysis; D007893:Leishmania donovani / Q000187:drug effects; D008081:Liposomes / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D051379:Mice; D021261:Parasitic Sensitivity Tests; D013329:Structure-Activity Relationship
chemical_list: D000935:Antifungal Agents; D008081:Liposomes; D000666:Amphotericin B