Development of Dual Efficacy Therapeutic Against Cryptosporidiosis and Giardiasis

Project Summary/AbstractThe potential for devastating consequences of cryptosporidiosis or giardiasis diarrhea in immunocompromisedpatients and malnourished children emphasizes the need for an effective therapy that could be usedsyndromically as a single agent appropriate for administration where diagnosis may be delayed or uncertain.This is also true for asymptomatic presentations of both diseases that can be devastating. The etiologicparasites colonize and reproduce in the small intestines of mammalian hosts where they are associated withmicrovilli of the epithelial cells. Hence, potential for finding dual therapeutic is high based on commonanatomical site of infection. Protein kinase inhibitors have attracted considerable attention as potentialtherapeutics since a number of them have been released as drugs in recent years and many are in variousphases of clinical trials. In a semi High throughput screening of MMV Pathogen box, we demonstrated thatdual hitting molecules including kinase inhibitors with no obvious chemical similarity with any previouslycharacterized anti-parasite drugs offer new medicinal chemistry opportunities for development of effectivemulti-parasite therapeutics against cryptosporidiosis and giardiasis. This research proposal capitalizes on theunique preliminary findings to hypothesize that effective single agent therapy that could be used syndromicallyfor treatment of either cryptosporidiosis or giardiasis is possible. In Years 1-2 (R21 phase): we will screen for,select and optimize dual effective hits to early leads. In Specific Aim 1a ? we will determine in vitro efficacy of~2000 unique chemical scaffold inhibitor clusters in blocking parasites growth using direct phenotypicscreening. We will select up to 50 hit scaffolds by rank order of efficacy against parasites and potential formammalian cytotoxicity. In Specific Aim 1b ? we will screen analogs of each of the 50 hit scaffolds identifiedin Aim 1a to develop structural activity relationship. Criteria for moving forward will include parasite killingefficiency, either static vs. cidal, selectivity, and ease of medicinal chemistry development. In Specific Aim 2we will use stability in simulated gastric/intestinal fluids, pharmacologic and pharmacokinetic studies to identify~20 effective early lead for both species and confirm clinical efficacy in mouse infection models. In Years 3-5(R33 phase): We will do lead optimization for efficacy, selectivity, and PK/ADMET properties under SpecificAim 3 - Medicinal chemistry; pharmacokinetic studies, in vitro compound efficacy, pharmacology, toxicity andresistance. In the final part of the project under Specific Aim 4 - In vivo compound efficacy and resistance,optimized leads will be evaluated for clinical efficacy in the mouse infection models and determine potentialease of developing acquired resistance and fitness cost of resistance to compounds being investigated. Theproduct of this research study would be useful as parasitic diarrheal therapeutic with the potential for broadspectrum activity. It will have significant public health benefits in many developing regions of the world.