sample="quota" bates="88197622" isource="ll" decade="1980" class="ui" date="19851016" MULTIDOSE RANGE-FINDING STUDY IN RATS B 180 FINAL REPORT MULTIDOSE RANGE-FINDING STUDY IN RATS B180 FINAL REPORT Submitted to Lorillard Greensboro, North Carolina October 16, 1985 HAZLETON LABORATORIES AMERICA, INC. 9200 LEESBURG TURNPIKE VIENNA VIRGINIA 22180 USA OFFICE OF QUALITY ASSURANCE Project Title: Multidose Range-Finding Study in Rats Project No.: 642-285 Quality Assurance review of the final report was conducted according to the procedures described in the standard operating procedures of the Report Review Section of the Office of Quality Assurance, and according to the general requirements of the Good Laboratory Practice regulations that were issued on December 22, 1978, by the Food and Drug Administration for compliance on and after June 20, 1979. The final report review was conducted and the findings were reported to management and to the study director on the following dates: Final Report Review 9/26/85 Findings Reported 9/30/85 Reviewer D. Hitzelberg Frederick G. Snyder Director Office of Quality Assurance SUBJECT: Multidose Range-Finding Study in Rats Project No. 642-285 We, the undersigned, hereby declare that the work was performed under our supervision, according to the procedures herein described. Study Director: Diplomate, American Board of Toxicology Life Sciences Division Laboratory Supervision: PRESTON L. BURLEW, B.A. Life Sciences Division Study Coordinator: JULIE A. RALPH, A.A.S. Life Sciences Division SPONSOR: Lorillard DATE: October 16, 1985 MATERIAL: B180 SUBJECT: FINAL REPORT Multidose Range-Finding Study in Rats Project No. 642-285 SUMMARY The test material, B180, was evaluated for the maximum tolerated dose (MTD) level in male and female rats. Based upon the findings of this study, the MTD in male and female rats was estimated to be 625 mg/kg. INTRODUCTION This study was designed to determine the maximum tolerated dose (MTD) level of B180 following repeated daily oral doses to rats for four days. The Single Dose Range-Finding Study was initiated on July 31, 1985 and was terminated August 4, 1985. The Multidose Range-Finding Study was initiated on August 21, 1985 and was terminated on August 25, 1985. TEST MATERIAL The test material, B180, a pale yellow liquid, was received from the sponsor on June 19, 1985, and was stored in an amber jar under refrigeration. Corn oil (Duke's® Pure Corn Oil, The C. F. Sauer Company, Lot No. 52500), a yellow liquid stored at room temperature, was received on April 30, 1985, and was used as the vehicle. A value of 100% active ingredient was assumed for purposes of dosage calculations. Information on stability and methods of synthesis, as well as data on composition or other characteristics which define the test material are on file with the sponsor. TEST ANIMALS Sprague-Dawley rats were received from Charles River Breeding Laboratories, Inc., Kingston, New York. Animals were randomly housed upon receipt via computer generated random numbers and subsequently assigned to this study following the acclimation period. Two rats of each sex were assigned to the Single Dose Range-Finding Study and six rats of each sex to the Multidose Range-Finding Study and subsequently to groups. The initial body weights of the males ranged from 190.7 to 214.9 grams, and the initial body weights of the females ranged from 144.0 to 159.1 grams in the Single Dose Range-Finding Study. In the Multidose Range-Finding Study, the initial body weights of the males ranged from 105.4 to 161.5 grams, and the initial body weights of the females ranged from 114.8 to 137.7 grams. The rats were identified uniquely by ear tags and housed individually in elevated wire-mesh cages. Commercial rodent ration (Purina Rodent Laboratory Chow® #5001) and tap water were available ad libitum. During the Single Dose Range-Finding Study, the temperature in the animal room was 72º and the humidity ranged from 60-78%. In the Multidose Range-Finding Study, the temperature in the animal room was 74ºF and the humidity ranged from 61-72%. A 12-hour light/dark cycle was maintained daily. The rats were acclimated to laboratory conditions for a minimum of one week prior to initiation of treatment. Rats were used in this study because they have historically been used in safety evaluation studies and are required by appropriate regulatory agencies. METHODS Groups and Dosage Levels Single Dose Range-Finding Study Two animals of each sex were assigned to the following group. Group 1 No of Animals Males 2 Females 2 Dosage Level mg/kg 5000 Multidose Range-Finding Study Two animals of each sex were assigned to each of the following groups. Compound Preparation and Administration The required amount of compound for each level was weighed into pre-calibrated glass beakers using an appropriate electronic balance. Corn oil was added to each beaker to bring the mixture to the desired volume. The mixtures were stirred for approximately five minutes on a magnetic stirrer and were stirred while dosing. During the Multidose Range-Finding phase, the test material mixtures were stored under refrigeration and brought to room temperature before dosing. Each rat received the appropriate amount of test material by gavage. Single Dose Range-Finding animals received one dose and Multidose Range-Finding animals received one dose daily for four consecutive days. The test material was administered by gavage because of the characteristics of the test compound. Observations and Records Single Dose Range-Finding Study All of the rats were observed for signs of toxic and pharmacologic effects at one and six hour postdose, and at least once daily thereafter to Day 5. Mortality/moribundity was recorded twice daily. Individual body weights were recorded immediately prior to dosing. Multidose Range-Finding Study All of the rats were observed for signs of toxic and pharmacologic effects at one and four hours postdose on Day 1, twice daily on Days 2 and 3, at one and six hours postdose on Day 4 and on Day 5 (24 hours following the day 4 dose). Mortality/morbundity was recorded twice daily. Individual body weights were recorded immediately prior to the first dose. Sacrifice and Gross Pathology Single Dose Range-Finding Study At termination (Day 5), all surviving rats were sacrificed by carbon dioxicde asphyxiation and discarded. Animals which died on study were also discarded. Multidose Range-Finding Study At termination (Day 5), all surviving rats were sacrificed by carbon dioxide asphyxiation and discarded. Raw Data and Final Report Storage All raw data and the final report are stored in the archives of Hazleton Laboratories America, Inc. Results Dosing Values Individual dosing volumes for the Single Dose and Multidose Range-Finding Studies are presented in Tables 1A and 1B, respectively. Mortality Single Dose Range-Finding Study Deaths occurring in the animals dosed at 5000 mg/kg consisted of one female rat on Day 1, six hours postdose. Multidose Range-Finding Study No deaths occurred during the Multidose Range-Finding Study. Clinical Observations and Body Weights Single Dose Range-Finding Study A summary of clinical observations is presented in Table 2A. Initial body weights are presented in Table 3A. A variety of commonly noted clinical signs was observed in both male and female rats during the study. Multidose Range-Finding Study A summary of clinical observations is presented in Table 2B. Initial body weights are presented in Table 3B. A variety of commonly noted clinical signs was observed in the Groups 2 and 3 males and in the Groups 1-3 females, with the severity increasing with increasing dose. The Group 1 males appeared normal throughout the study. Conclusions Based upon the presence of two incidences of gross toxicity (slight depression and/or depression) in both males and females dosed at 1000 mg/kg of body weight, the MTD was estimated to be 625 mg/kg of body weight in male and female rats.