title: Preclinical evaluation of injectable reduced hydroxocobalamin as an antidote to acute carbon monoxide poisoning.
abstract: BACKGROUND
  Current management of acute inhalational carbon monoxide (CO) toxicity includes hyperbaric or normobaric O2 therapy. However, efficacy has not been established. The purpose of this study was to establish therapeutic proof of concept for a novel injectable antidote consisting of the combination of hydroxocobalamin and ascorbic acid into a reduced form (B12r) as demonstrated by clinically significant increase (>500 ppm) in CO2 production, reduced carboxyhemoglobin (COHgb) half-life (COHgb t1/2), and increased cerebral O2 delivery and attenuation of CO-induced microglial damage in a preclinical rodent model of CO toxicity.
  
  
  METHODS
  B12r-mediated conversion of CO to CO2 and COHgb t1/2 in human blood were measured by gas analysis and Raman resonance spectroscopy. Rats were exposed to either air or CO and then injected with saline or B12r. Cognitive assessment was tested in a Morris water maze. Brain oxygenation was measured with Licox. Brain histology was assessed by fluorescent antibody markers and cell counts.
  
  
  RESULTS
  B12r resulted in significant CO2 production (1,170 ppm), compared with controls. COHgb t1/2 was reduced from 33 minutes (normal saline) to 17.5 (p < 0.001). In rat models, severe CO-induced brain hypoxia (PbtO2, 18 mm Hg) was followed by significant reduction in τ25 to 12 minutes for B12r rats versus 40 minutes for normal saline-treated rats (p < 0.0001). There was major attenuation of CO-induced microglial damage, although cognitive performance differences were minimal.
  
  
  CONCLUSIONS
  Our preclinical data suggest that the novel synergism of hydroxocobalamin with ascorbic acid has the potential to extract CO through conversion to CO2, independently of high-flow or high-pressure O2. This resulted in a clinically significant off-gassing of CO2 at levels five to eight times greater than those of controls, a clinically significant reduction in COHgb half-life, and evidence of increased brain oxygenation and amelioration of myoglial damage in rat models. Reduced hydroxocobalamin has major potential as an injectable antidote for CO toxicity.
authors.affiliation: From the Departments of Physiology (J.D.R., C.S.J.), Anesthesiology (P.S.R., B.D.), and Biology (L.G.S.), Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Roderique
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Josef
authors.forename: Alden H
authors.initials: AH
authors.lastname: Newcomb
authors.forename: Penny S
authors.initials: PS
authors.lastname: Reynolds
authors.forename: Leonardo G
authors.initials: LG
authors.lastname: Somera
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Spiess
mesh_terms: D000818:Animals; D000931:Antidotes / Q000494:pharmacology*; D001205:Ascorbic Acid / Q000494:pharmacology*; D002249:Carbon Monoxide Poisoning / Q000188:drug therapy*; D006801:Humans; D006879:Hydroxocobalamin / Q000494:pharmacology*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D018782:Maze Learning; D018613:Microscopy, Confocal; D010102:Oxygen Inhalation Therapy; D051381:Rats; D020318:Rats, Long-Evans; D017207:Rats, Sprague-Dawley; D013059:Spectrum Analysis, Raman
chemical_list: D000931:Antidotes; D001205:Ascorbic Acid; D006879:Hydroxocobalamin