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Title: In vivo screening of candidate pretreatment compounds against cyanide using mice

Abstract

An in vivo screening procedure was established at Battelle's Medical Research and Evaluation Facility (MREF) to evaluate the efficacy of candidate pretreatment compounds in mice challenged with the blood agent, sodium cyanide (NaCN). Male albino mice of ICR outbred stock weighing between 22.5 and 27.5 g are challenged by intramuscular (i.m.) injection, at a volume of 0.5 mL/kg, of a dose of NaCN twice the LD50 of untreated mice as determined on that day of testing. Candidate drugs are tested at fractions of their LD50 or their limit of solubility in the most optimum vehicle and given intraperitoneally (i.p.) to separate groups of mice at either 60 or 15 min prior to NaCN challenge. Sodium thiosulfate (1000 mg/kg)/sodium nitrite (100 mg/kg) controls are injected i.p. only at 60 min prior to challenge. A test compound is deemed effective if, at any of three concentrations tested, or at either pretreatment time, it is statistically more efficacious in preventing lethality than is a negative control substance (candidate compound vehicle).

Authors:
; ; ; ;
Publication Date:
Research Org.:
Battelle Memorial Inst., Columbus, OH (United States)
OSTI Identifier:
7043512
Report Number(s):
AD-P-008846/8/XAB
CNN: DAMD17-83-C-3129
Resource Type:
Technical Report
Resource Relation:
Other Information: This article is from 'Proceedings of the Medical Defense Bioscience Review (1993) Held in Baltimore, Maryland on 10-13 May 1993. Volume 2', AD-A275 668, 911-919
Country of Publication:
United States
Language:
English
Subject:
45 MILITARY TECHNOLOGY, WEAPONRY, AND NATIONAL DEFENSE; 63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; 59 BASIC BIOLOGICAL SCIENCES; CHEMICAL WARFARE AGENTS; PREVENTIVE MEDICINE; CYANIDES; LETHAL DOSES; DOSES; MEDICINE; WEAPONS 450600* -- Military Technology, Weaponry, & National Defense-- Chemical & Biological-- (1990); 560300 -- Chemicals Metabolism & Toxicology; 550200 -- Biochemistry

Citation Formats

Kiser, R.C., Olson, C.T., Menton, R.G., Hobson, D.W., and Moore, D.M.. In vivo screening of candidate pretreatment compounds against cyanide using mice. United States: N. p., 1993. Web.
Kiser, R.C., Olson, C.T., Menton, R.G., Hobson, D.W., & Moore, D.M.. In vivo screening of candidate pretreatment compounds against cyanide using mice. United States.
Kiser, R.C., Olson, C.T., Menton, R.G., Hobson, D.W., and Moore, D.M.. 1993. "In vivo screening of candidate pretreatment compounds against cyanide using mice". United States. doi:.
@article{osti_7043512,
title = {In vivo screening of candidate pretreatment compounds against cyanide using mice},
author = {Kiser, R.C. and Olson, C.T. and Menton, R.G. and Hobson, D.W. and Moore, D.M.},
abstractNote = {An in vivo screening procedure was established at Battelle's Medical Research and Evaluation Facility (MREF) to evaluate the efficacy of candidate pretreatment compounds in mice challenged with the blood agent, sodium cyanide (NaCN). Male albino mice of ICR outbred stock weighing between 22.5 and 27.5 g are challenged by intramuscular (i.m.) injection, at a volume of 0.5 mL/kg, of a dose of NaCN twice the LD50 of untreated mice as determined on that day of testing. Candidate drugs are tested at fractions of their LD50 or their limit of solubility in the most optimum vehicle and given intraperitoneally (i.p.) to separate groups of mice at either 60 or 15 min prior to NaCN challenge. Sodium thiosulfate (1000 mg/kg)/sodium nitrite (100 mg/kg) controls are injected i.p. only at 60 min prior to challenge. A test compound is deemed effective if, at any of three concentrations tested, or at either pretreatment time, it is statistically more efficacious in preventing lethality than is a negative control substance (candidate compound vehicle).},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = 1993,
month = 5
}

Technical Report:
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  • This task was a continuation of Task 86-29 initiated for Contract D. It provided in vivo screens for evaluating the efficacy of candidate pretreatment and treatment compounds submitted by the Drug Assessment Division of U.S. Army Medical Research of Chemical Defense against soman, tabun, and/or cyanide. A total of 578 compounds were received for testing and their maximum solubility in-vehicles comparable with in vivo testing in mice was determined. Range-finding and median lethal dose determinations following IM and/or oral administrations were conducted for 436 compounds submitted for nerve agent screening and range finding and median lethal dose determinations following IPmore » administration were conducted for up to 142 compounds submitted for cyanide screening. Of 332 compounds evaluated, 154 passed the GD treatment efficacy evaluation, and 90 of 156 compounds submitted passed the GA treatment efficacy evaluation. For pretreatment studies against a GD challenge, 224 of 379 compounds submitted passed the IM efficacy evaluation and 96 of 143 compounds submitted passed the oral efficacy evaluation. Only 12 of 133 compounds evaluated as cyanide pretreatment compounds passed the efficacy evaluations. The mission of Task 89-01 was combined under Task 91-20 for the duration of Contract DAMD17-89-C-9050.« less
  • An assay measuring propidium iodide (PI) incorporation into nonviable human peripheral blood mononuclear leukocytes (PBML) was established at the U.S. Army Medical Research Institute of Chemical Defense (USAMRICD), and the technology transferred and implemented at Battelle's Medical Research and Evaluation Facility (MREF) for use as a screen to evaluate candidate compounds for direct cytotoxicity as well as for efficacy in preventing HD-induced cytotoxicity. For assay transition, studies were performed to establish a fixed HD challenge concentration; to develop a positive and negative control dataset; and to establish the reproducibility in obtaining an EC50 (concentration of candidate compound required to providemore » 50 percent protection against the fixed HD concentration) for niacinamide (NM). Various concentrations of candidate compounds were preincubated for 15 to 30 min with PBML prior to adding the fixed HD challenge. At 24 hr after exposure, PI was added to the cultures and the number of nonviable (PI positive) cells was determined by flow cytometry. Positive (NM pretreated) and negative (HD only) controls were examined concurrently and used to maintain data quality. From this dataset, candidate compounds were evaluated for direct cytotoxic effects and for efficacy in preventing HD-induced cytotoxicity. EC50 values for effective candidate compounds were estimated and reported for ranking compound effectiveness. Results from these studies demonstrate assay function and reproducibility during routine screening operations.« less
  • A task was instituted at the Medical Research and Evaluation Facility (MREF) to develop in vitro assays to screen pretreatment and treatment compounds for their ability to protect or reverse the toxic effects of organophosphates and vesicants. Four vesicant assays and three nerve agent assays were developed. Two of the vesicant assays were for cell viability of keratinocyte, one in the presence of distilled mustard and one lewisite. One assay determines the effect of vesicants on keratinocyte reproduction and the other the effect of distilled mustard on cellular coenzyme nicotinamide adenine dinucleotide content. The organophosphate assays measure the effects onmore » acetylcholinesterase of selected compounds measured by ability to reactivate, effect on aging rate, and directly. In vitro screen; HD; L; Cellular NAD+ cellular viability; GA; GD; VX; Acetylcholinesterase inhibition; Reactivators; RA 5; Aging rate; Keratinocytes; Treatment and pretreatments; Assaying; Tabun (GA); Sarin (GB); Soman (GD); Organoarsenic; Organophosphates; Chemical Surety Material (CSM); Blisters; Toxicity; Toxic agents; Nerve agents; Chemical warfare agents; G Agents; V Agents; Vesicants; Mustard agents.« less
  • MREF Task 89-07 encompassed four vesicant assays and four nerve agent assays. The four vesicant assays evaluated the candidate P and T compound solubility limitations, direct cytotoxic effects, efficacy against HD-induced cellular nicotinamide adenine dinucleotide (NAD+) depletion, and efficacy against HD-induced cytotoxicity. Normal human epidermal cells (NHEKs) were used to evaluate candidate PT compound efficacy against HD-induced NAD+ depletion, and peripheral blood mononuclear leukocytes (PBMC) were used in direct cytotoxicity and HD-induced cytotoxicity assays. The four nerve agent assays assessed candidate PT compound direct inhibitory effects on acetylcholinesterase (AChE) activity, candidate PT compound efficacy in reactivating Tabun (GA) - andmore » O-ethyl S-(2-diisopropylaminoethyl) methylphosphonothiolate (VX)-inhibited A ThE, and candidate PT compound efficacy in slowing the aging rate of Soman (GD) inhibited AChE. All nerve agent and vesicant assays with the exception of the direct cytotoxicity and HD-induced cytotoxicity assays were initially established under MREF Task 88-36. The direct cytotoxicity and HD-induced cytotoxicity assays were transitioned to the MREF from USAMRICD and validated for use in routine screening procedures, including the generation of control database values, under Task 89-07. Solubility data were obtained for 37 compounds submitted for evaluation in the vesicant assays. Thirty-five of these compounds were evaluated for direct cytotoxicity, and their effect against HD-induced cytotoxicity, while 13 compound is were evaluated for efficacy against HD-induced NAD+ depletion. AChE reactivation, ACHE aging, ACHE inhibition, In vitro, Cytotoxicity , Vesicant assays, Nerve ag.« less
  • Huperzine A (HUP) is a naturally-occurring, potent, reversible inhibitor of acetylcholinesterase (AChE) that crosses the blood-brain barrier. To examine its ability to protect against nerve agent poisoning, HUP was administered i.p. to mice, and the s.c. LD50 of soman was determined at various time intervals after pretreatment. Results were compared to those obtained for animals treated with physostigmine. A protective ratio of approximately 2 was maintained for at least 6 hr after a single injection of HUP, without the need for any post-challenge drug therapy. By contrast, pretreatment with physostigmine increased the LD50 of soman by 1.4- to 1.5-fold formore » only up to 90 min. The long-lasting antidotal efficacy displayed by HUP correlated with the time course of the blood-AChE inhibition. The results suggest that the protection of animals by HUP from soman poisoning was achieved by temporarily sequestering the active site region of the physiologically important AChE.« less