title: Assessment of drug candidates for broad-spectrum antiviral therapy targeting cellular pyrimidine biosynthesis.
abstract: Currently available antiviral drugs frequently induce side-effects or selection of drug-resistant viruses. We describe a novel antiviral principle based on targeting the cellular enzyme dihydroorotate dehydrogenase (DHODH). In silico drug design and biochemical evaluation identified Compound 1 (Cmp1) as a selective inhibitor of human DHODH in vitro (IC50 1.5±0.2nM). Crystallization data specified the mode of drug-target interaction. Importantly, Cmp1 displayed a very potent antiviral activity that could be reversed by co-application of uridine or other pyrimidine precursors, underlining the postulated DHODH-directed mode of activity. Human and animal cytomegaloviruses as well as adenoviruses showed strong sensitivity towards Cmp1 in cell culture-based infection systems with IC50 values in the low micromolar to nanomolar range. Particularly, broad inhibitory activity was demonstrated for various types of laboratory and clinically relevant adenoviruses. For replication of human cytomegalovirus in primary fibroblasts, antiviral mode of activity was attributed to the early stage of gene expression. A mouse in vivo model proved reduced replication of murine cytomegalovirus in various organs upon Cmp1 treatment. These findings suggested Cmp1 as drug candidate and validated DHODH as a promising cellular target for antiviral therapy.
authors.affiliation: Institute for Clinical and Molecular Virology, University of Erlangen-Nuremberg, Erlangen, Germany. Electronic address: manfred.marschall@viro.med.uni-erlangen.de.
authors.forename: Manfred
authors.initials: M
authors.lastname: Marschall
authors.forename: Ina
authors.initials: I
authors.lastname: Niemann
authors.forename: Karin
authors.initials: K
authors.lastname: Kosulin
authors.forename: Anna
authors.initials: A
authors.lastname: Bootz
authors.forename: Sabrina
authors.initials: S
authors.lastname: Wagner
authors.forename: Thomas
authors.initials: T
authors.lastname: Dobner
authors.forename: Thomas
authors.initials: T
authors.lastname: Herz
authors.forename: Bernd
authors.initials: B
authors.lastname: Kramer
authors.forename: Johann
authors.initials: J
authors.lastname: Leban
authors.forename: Daniel
authors.initials: D
authors.lastname: Vitt
authors.forename: Thomas
authors.initials: T
authors.lastname: Stamminger
authors.forename: Corina
authors.initials: C
authors.lastname: Hutterer
authors.forename: Stefan
authors.initials: S
authors.lastname: Strobl
mesh_terms: D000260:Adenoviruses, Human / Q000187:drug effects; D000818:Animals; D000963:Antimetabolites / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D000998:Antiviral Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D002478:Cells, Cultured; D003198:Computer Simulation; D003587:Cytomegalovirus / Q000187:drug effects; D000090722:Dihydroorotate Dehydrogenase; D015195:Drug Design; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology / Q000821:virology; D015774:Ganciclovir / Q000494:pharmacology; D006564:Herpesviridae / Q000187:drug effects; D006801:Humans; D020128:Inhibitory Concentration 50; D051379:Mice; D008958:Models, Molecular; D015394:Molecular Structure; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000037:antagonists & inhibitors*; D011743:Pyrimidines / Q000096:biosynthesis*; D013047:Specific Pathogen-Free Organisms; D013329:Structure-Activity Relationship; D014616:Vaccinia virus / Q000187:drug effects; D014776:Virus Cultivation
chemical_list: D000963:Antimetabolites; D000998:Antiviral Agents; D000090722:Dihydroorotate Dehydrogenase; D004791:Enzyme Inhibitors; D011743:Pyrimidines; D044925:Oxidoreductases Acting on CH-CH Group Donors; D015774:Ganciclovir
keywords: 4SCan drug design; Antiviral drug candidate; Broad-spectrum antiviral therapy; Cytomegaloviruses and adenoviruses; Pyrimidine biosynthesis; Target dihydroorotate dehydrogenase