Research Projects

Mucormycosis, an invasive fungal infection, has a very high mortality rate (50%) due to the inability of the human immune system to eradicate the underlying pathogen, particularly in immunocompromised individuals, as recently observed among COVID-19 patients [1]. Centers for Disease Control and Prevention, United States and other global guidelines strongly recommend the use of high doses (5–10 mg/kg per day) of Amphotericin B (AMB), a polyene macrolide antibiotic, as the first-line treatment for mucormycosis [2]. However, AMB, despite being highly efficacious against mucormycosis, is associated with severe dose-dependent and duration-dependent nephrotoxicity [3], which further leads to acute renal failure in more than 50 % of the individuals [4]. N-acetylcysteine (NAC), a mucolytic agent, when co-administered with AMB through oral as well as parenteral routes, has been reported to provide a significant reduction in the nephrotoxic effect of AMB, but leads to gastrointestinal discomfort in some patients [5, 6]. In contrast, no studies have been reported so far highlighting the role of the co-delivery of NAC in the presence of AMB through the inhalational route that can potentiate the antifungal action, along with eliminating the nephrotoxicity of AMB and minimizing gastrointestinal discomfort of NAC due to their limited systemic exposure. The proposed research project is aimed at treatment of population suffering from mucormycosis (P) by developing inhalable spray-dried lyotropic liquid crystals of AMB and NAC (AMB-NAC-LLC) that can be delivered directly to the lungs by self-administration through dry powder inhalers (DPIs) (I) to minimize the nephrotoxicity and enhance the antifungal activity of Amphotericin B (AMB) (O) which is very high in case of the currently available marketed parenteral formulations of AMB administered as slow intravenous infusion due to their direct systemic exposure (C). AMB-NAC-LLC will enable direct delivery of AMB at the infected regions due to the bioadhesive spray-dried particles in the mass median aerodynamic diameter (MMAD) range of the inhalable formulations, i.e. 1-5 µm [7]. Ultimately, this leads to lower renal exposure. Moreover, the co-administration of NAC with AMB will offer enhanced penetration of AMB into the infected tissues, as well as direct clearance of the fungal conidia due to its mucolytic, expectorant action on the thick and viscid mucus produced excessively by the natural defence system of the human body to counter foreign agents [8, 9]. The novelty of the proposed project lies in the fact that this will be the first study to explore the development and preclinical evaluation of inhalable spray-dried lyotropic liquid crystals of AMB and NAC (AMB-NAC-LLC) for mucormycosis (black fungus). For this purpose, a quality by design approach will be followed for the development of AMB-NAC-LLC employing spray drying technology; thereafter, multidimensional evaluation (in vitro/in vivo) of the AMB-NAC-LLC will be performed as per the approved protocol. The properties to be evaluated are in vitro aerosolization properties, antifungal activity, drug release characteristics, and in vitro cytotoxicity on pulmonary epithelial cells, followed by in vivo pharmacokinetics along with organ/tissue distribution studies and pharmacodynamic studies under a controlled environment in a Biosafety Facility (Level-2). Furthermore, PK/PD modelling will be explored for the dose prediction of AMB-NAC-LLC for its use among humans. The prepared AMB-NAC-LLC are expected to: (a) Be safe for clinical use, as all the ingredients and raw materials taken will be from the Generally Recognized as Safe (GRAS) category as per USFDA; (b) Be an industrial viable product because it will be developed following the guidelines of Good Manufacturing Practices (GMP); and (c) Establish preclinical evidence related to the safety and efficacy of AMB-NAC-LLC so that it can be considered for clinical trials as soon as possible.