title: A nanotherapy strategy significantly enhances anticryptosporidial activity of an inhibitor of bifunctional thymidylate synthase-dihydrofolate reductase from Cryptosporidium.
abstract: Cryptosporidiosis, a gastrointestinal disease caused by protozoans of the genus Cryptosporidium, is a common cause of diarrheal diseases and often fatal in immunocompromised individuals. Bifunctional thymidylate synthase-dihydrofolate reductase (TS-DHFR) from Cryptosporidium hominis (C. hominis) has been a molecular target for inhibitor design. C. hominis TS-DHFR inhibitors with nM potency at a biochemical level have been developed however drug delivery to achieve comparable antiparasitic activity in Cryptosporidium infected cell culture has been a major hurdle for designing effective therapies. Previous mechanistic and structural studies have identified compound 906 as a nM C. hominis TS-DHFR inhibitor in vitro, having μM antiparasitic activity in cell culture. In this work, proof of concept studies are presented using a nanotherapy approach to improve drug delivery and the antiparasitic activity of 906 in cell culture. We utilized PLGA nanoparticles that were loaded with 906 (NP-906) and conjugated with antibodies to the Cryptosporidium specific protein, CP2, on the nanoparticle surface in order to specifically target the parasite. Our results indicate that CP2 labeled NP-906 (CP2-NP-906) reduces the level of parasites by 200-fold in cell culture, while NP-906 resulted in 4.4-fold decrease. Moreover, the anticryptosporidial potency of 906 improved 15 to 78-fold confirming the utility of the antibody conjugated nanoparticles as an effective drug delivery strategy.
authors.affiliation: Department of Molecular Biology & Immunology and Institute for Cancer Research, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, Fort Worth, TX 76107, USA.
authors.forename: Anindita
authors.initials: A
authors.lastname: Mukerjee
authors.affiliation: Department of Pharmacology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
authors.forename: Pinar
authors.initials: P
authors.lastname: Iyidogan
authors.affiliation: Infectious Disease Division, Department of Internal Medicine, University of Texas Medical Branch, Galveston, TX 77555, USA.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Castellanos-Gonzalez
authors.affiliation: Department of Chemistry, Yale University, 225 Prospect Street, PO Box 208107, New Haven, CT 06520, USA.
authors.forename: José A
authors.initials: JA
authors.lastname: Cisneros
authors.affiliation: Department of Pharmacology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Czyzyk
authors.affiliation: Department of Molecular Biology & Immunology and Institute for Cancer Research, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, Fort Worth, TX 76107, USA.
authors.forename: Amalendu Prakash
authors.initials: AP
authors.lastname: Ranjan
authors.affiliation: Department of Chemistry, Yale University, 225 Prospect Street, PO Box 208107, New Haven, CT 06520, USA.
authors.forename: William L
authors.initials: WL
authors.lastname: Jorgensen
authors.affiliation: Infectious Disease Division, Department of Internal Medicine, University of Texas Medical Branch, Galveston, TX 77555, USA.
authors.forename: A Clinton
authors.initials: AC
authors.lastname: White
authors.affiliation: Department of Molecular Biology & Immunology and Institute for Cancer Research, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, Fort Worth, TX 76107, USA.
authors.forename: Jamboor K
authors.initials: JK
authors.lastname: Vishwanatha
authors.affiliation: Department of Pharmacology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA. Electronic address: karen.anderson@yale.edu.
authors.forename: Karen S
authors.initials: KS
authors.lastname: Anderson
mesh_terms: D000981:Antiprotozoal Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D001665:Binding Sites; D002478:Cells, Cultured; D003458:Cryptosporidium / Q000187:drug effects* / Q000201:enzymology*; D004357:Drug Synergism; D008958:Models, Molecular; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors*; D053758:Nanoparticles / Q000737:chemistry*; D013762:Tetrahydrofolate Dehydrogenase; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors*
chemical_list: D000981:Antiprotozoal Agents; D009097:Multienzyme Complexes; C056930:thymidylate synthase-dihydrofolate reductase; D013762:Tetrahydrofolate Dehydrogenase; D013940:Thymidylate Synthase
keywords: Cryptosporidium hominis; Dihydrofolate reductase; Drug delivery; Nanoparticle; Thymidylate synthase