Drug Discovery: Supporting development of new drugs to treat global parasitic diseases
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1 Drug Discovery: Supporting development of new drugs to treat global parasitic diseases UC Santa Cruz Bio 117 Feb. 23, 2016 Judy Sakanari Center for Parasitic Diseases UC San Francisco
2 Parasitic Diseases, Worldwide Malaria (in 2012) >200 mill infected >600,000 deaths Schistosomiasis (2014) Chagas Disease (2008) Leishmaniasis (2014) Sleeping Sickness (2012) River Blindness (2014) Lymphatic filariasis (2014) >200 million infected 200,000 deaths 8 mill infected 11,000 deaths 1.3 mill infected 20-30,000 deaths 30,000 infected 9,000 deaths 18 mill infected 270,000 blind 40 mill disfigured Soil-transmitted helminths (2014) >1.5 billion infected
3
4 Onchocerciasis (River Blindness) Filarial nematode: Onchocerca volvulus Regions affected: 36 countries in Sub-Saharan Africa, foci in Americas Number infected: 36 million, 270,000 are blind; 1.2 million visually impaired Number at risk: 120 million worldwide Clare Gilbert. International Centre for Eye Health LSHTM
5 Simulium damnosum, the blackly vector Transmitted by the blackfly microfilariae in skin Microfilariae migrate throughout the skin and are ingested by adult black flies when the flies take a blood meal. Larval black flies inhabit fastflowing rivers, hence the name River Blindness.
6 Symptoms of intense itchiness; conditions of loss of skin integrity and blindness.
7 video/default.aspx? youtube_id=nzh4aobqk0y&c ategory=&filter=river %20blindness
8 Macrofilarial worms Onchocerca volvulus Brugia & Wuchereria Loa loa Drugs to treat the diseases Loa loa (eyeworm) Ivermectin, which kills the microfilariae, is used in the treatment of river blindness and lymphatic filariasis. When patients also have a high number of Loa loa microfilariae, the drug can cause severe adverse reactions and fatalities.
9 2015 Nobel Prize in Physiology or Medicine Tu Youyou Satoshi Omura William C. Campbell Tu discovered Artemisinin, a drug used to treat malaria. Omura and Campbell developed Avermectin (Ivermectin) for treatment of parasitic worm infections.
10 Project Goals To identify macrofilaricidal drugs for onchocerciasis in Loa loa endemic areas and for the treatment of lymphatic filariasis.
11 Compound libraries Macrofilaria Project In vitro potency Physiochemical properties and in vivo studies U.C. San Francisco Brugia adult worms in vitro screens University of Buea, Cameroon Onchocerca ochengi in cattle in vitro assays adult worms, mf Loa loa mf in vitro assays In vitro studies ADME, stability, solubility, permeability, cell toxicity Med Chemistry U.C. San Francisco animal model of infection NY Blood Center Onchocerca volvulus larval molting assay Univ. Buea, Cameroon animal model with O. ochengi Preclinical Candidates
12 Biomek media removal and compound dispensing Four 24-well plates/20 mins. (1,000 worms take ~5 hrs. 250 cmpds)
13 The Worminator quantifies worm movement Marcellino C, Gut J, Singh R, McKerrow, J and Sakanari J WormAssay: A Novel Computer Application for Whole-Plate Motion-based Screening of Macroscopic Parasites. PLoS Negl Trop Dis 6(1): e1494.
14 Screening Onchocerca ochengi adults: motility and viability S. Lustigman, NY Blood Center; F. Cho-Ngwa, M. Samje, Univ. Buea, Cameroon 7 day assay: killing determined using MTT staining and visual observation for color change blue = live no color = dead
15 Onchocerca volvulus L3 molting assay cast from the molt Sara Lus(gman, NY Blood Center
16 Screening Funnel 2,000 FDA-approved drugs were screened using the Worminator assay with adult Brugia and Onchocerca ochengi worms in in vitro assays and larval O. volvulus in the molting assay.
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18 Auranofin IC50s in vitro assays Brugia pahangi females Day 3 B. pahangi males Day 3 Onchocerca ochengi females Day 7 O. ochengi males Day 5 O. volvulus L3 Day 6 O. ochengi mf Day 5 Loa loa mf Day um um um um um 3 um 12.8 um ~50x
19 TEM of female worms from in vitro worm assay Day 1 1% DMSO 8% inhibi(on of mo(lity 1 um Auranofin 96% inhibi(on of mo(lity Bulman et al., PLoS NTD 2015
20 Auranofin as an antiparasitic agent Protozoa Entamoeba histolytica Giardia intestinalis Cryptosporidium Plasmodium falciparum Leishmania infantum, L. major Trypanosoma brucei Helminths Echinococcus granulosus Taenia crassiceps Schistosoma mansoni Haemonchus contortus Brugia, Onchocerca Thioredoxin reductase systems are important in preventing oxidative damage due to oxygen metabolism; the parasite s enzyme is a good target for an antiparasitic drug.
21 Low-hanging fruit: Auranofin FDA-approved Auranofin is a gold salt used in treating rheumatoid arthritis for >25 years. Single oral 3mg/kg/day for 7 days decreased liver damage in hamsters infected with Entamoeba histolytica (Debnath et al. Nature Med 2012). Human plasma elimination half-life with auranofin is days (Blodgett et al. 1984). NIH/NIAID human study: Evaluation of PK and PD of Auranofin for Short Course Treatment of Amebiasis evaluate safety of auranofin, once/day oral dosing for 7 days in healthy adult subjects. 3 mg capsule Auranofin (Brand name: Ridaura)
22 Summary Worminator is used for in vitro screening of macroparasites and microparasites. Auranofin identified from FDA library as a lead candidate. Filarial thioredoxin reductase may be target of auranofin. Biochemical and structural studies of the TxR target. Storey et al IJPDDR
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