title: Drug-Based Lead Discovery: The Novel Ablative Antiretroviral Profile of Deferiprone in HIV-1-Infected Cells and in HIV-Infected Treatment-Naive Subjects of a Double-Blind, Placebo-Controlled, Randomized Exploratory Trial.
abstract: Antiretrovirals suppress HIV-1 production yet spare the sites of HIV-1 production, the HIV-1 DNA-harboring cells that evade immune detection and enable viral resistance on-drug and viral rebound off-drug. Therapeutic ablation of pathogenic cells markedly improves the outcome of many diseases. We extend this strategy to HIV-1 infection. Using drug-based lead discovery, we report the concentration threshold-dependent antiretroviral action of the medicinal chelator deferiprone and validate preclinical findings by a proof-of-concept double-blind trial. In isolate-infected primary cultures, supra-threshold concentrations during deferiprone monotherapy caused decline of HIV-1 RNA and HIV-1 DNA; did not allow viral breakthrough for up to 35 days on-drug, indicating resiliency against viral resistance; and prevented, for at least 87 days off-drug, viral rebound. Displaying a steep dose-effect curve, deferiprone produced infection-independent deficiency of hydroxylated hypusyl-eIF5A. However, unhydroxylated deoxyhypusyl-eIF5A accumulated particularly in HIV-infected cells; they preferentially underwent apoptotic DNA fragmentation. Since the threshold, ascertained at about 150 μM, is achievable in deferiprone-treated patients, we proceeded from cell culture directly to an exploratory trial. HIV-1 RNA was measured after 7 days on-drug and after 28 and 56 days off-drug. Subjects who attained supra-threshold concentrations in serum and completed the protocol of 17 oral doses, experienced a zidovudine-like decline of HIV-1 RNA on-drug that was maintained off-drug without statistically significant rebound for 8 weeks, over 670 times the drug's half-life and thus clearance from circulation. The uniform deferiprone threshold is in agreement with mapping of, and crystallographic 3D-data on, the active site of deoxyhypusyl hydroxylase (DOHH), the eIF5A-hydroxylating enzyme. We propose that deficiency of hypusine-containing eIF5A impedes the translation of mRNAs encoding proline cluster ('polyproline')-containing proteins, exemplified by Gag/p24, and facilitated by the excess of deoxyhypusine-containing eIF5A, releases the innate apoptotic defense of HIV-infected cells from viral blockade, thus depleting the cellular reservoir of HIV-1 DNA that drives breakthrough and rebound.
  
  
  
  ClinicalTrial.gov NCT02191657.
authors.affiliation: Department of Pediatrics, New Jersey Medical School, Rutgers University, Newark, New Jersey, United States of America.
authors.forename: Deepti
authors.initials: D
authors.lastname: Saxena
authors.affiliation: Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario, Canada.|ApoPharma Inc., Toronto, Ontario, Canada.
authors.forename: Michael
authors.initials: M
authors.lastname: Spino
authors.affiliation: ApoPharma Inc., Toronto, Ontario, Canada.
authors.forename: Fernando
authors.initials: F
authors.lastname: Tricta
authors.affiliation: ApoPharma Inc., Toronto, Ontario, Canada.
authors.forename: John
authors.initials: J
authors.lastname: Connelly
authors.affiliation: Department of Obstetrics, Gynecology and Women's Health, New Jersey Medical School, Rutgers University, Newark, New Jersey, United States of America.
authors.forename: Bernadette M
authors.initials: BM
authors.lastname: Cracchiolo
authors.affiliation: Oncology Center and Medical Clinic III, Asklepios Klinik St. Georg, Hamburg, Germany.
authors.forename: Axel-Rainer
authors.initials: AR
authors.lastname: Hanauske
authors.affiliation: Department of Chemistry, Manhattanville College, Purchase, New York, United States of America.
authors.forename: Darlene
authors.initials: D
authors.lastname: D'Alliessi Gandolfi
authors.affiliation: Department of Medicine, New Jersey Medical School, Rutgers University, Newark, New Jersey, United States of America.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Mathews
authors.affiliation: Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, Cleveland, Ohio, United States of America.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Karn
authors.affiliation: Department of Medicine, New Jersey Medical School, Rutgers University, Newark, New Jersey, United States of America.
authors.forename: Bart
authors.initials: B
authors.lastname: Holland
authors.affiliation: Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institute of Health, Bethesda, Maryland, United States of America.
authors.forename: Myung Hee
authors.initials: MH
authors.lastname: Park
authors.affiliation: Department of Medicine, New Jersey Medical School, Rutgers University, Newark, New Jersey, United States of America.
authors.forename: Tsafi
authors.initials: T
authors.lastname: Pe'ery
authors.affiliation: Department of Pediatrics, New Jersey Medical School, Rutgers University, Newark, New Jersey, United States of America.
authors.forename: Paul E
authors.initials: PE
authors.lastname: Palumbo
authors.affiliation: Department of Pediatrics, New Jersey Medical School, Rutgers University, Newark, New Jersey, United States of America.|Department of Obstetrics, Gynecology and Women's Health, New Jersey Medical School, Rutgers University, Newark, New Jersey, United States of America.|Department of Microbiology, Biochemistry and Molecular Genetics, New Jersey Medical School, Rutgers University, Newark, New Jersey, United States of America.
authors.forename: Hartmut M
authors.initials: HM
authors.lastname: Hanauske-Abel
mesh_terms: D000293:Adolescent; D000328:Adult; D019380:Anti-HIV Agents / Q000008:administration & dosage / Q000009:adverse effects / Q000494:pharmacology / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002478:Cells, Cultured; D000077543:Deferiprone; D004305:Dose-Response Relationship, Drug; D004311:Double-Blind Method; D055808:Drug Discovery; D005260:Female; D015658:HIV Infections / Q000188:drug therapy*; D015497:HIV-1 / Q000187:drug effects; D006801:Humans; D008297:Male; D008875:Middle Aged; D011728:Pyridones / Q000008:administration & dosage / Q000009:adverse effects / Q000494:pharmacology / Q000627:therapeutic use*
chemical_list: D019380:Anti-HIV Agents; D011728:Pyridones; D000077543:Deferiprone