Document yknY1pN4Mw4O502GvdZMm72DX

SPONSOR ElfAtochemS.A. Cours Michelet ' La Defense 10 92091 Paris-la-Defense CEDEX France STUDY TITLE ACUTE ORAL TOXICITY IN RATS f TEST SUBSTANCE STUDY DIRECTOR Xavier Manciaux CONFIDENTIAL AR226-3096 STUDY COMPLETION DATE 14 May 1998 PERFORMING LABORATORY Centre International de Toxicologie (C.I.T.) Miserey - 27005 Evreux - France LABORATORY STUDY NUMBER 16045 TAR Company Sanitized. Does not contain TSFA S^W ^IX/OLUU^ 11U. lUU'-t-J ^ ^r<JI----------fJUM^Mdl /-UUl-llCIH O..ft.. CONTENTS STATEMENT OF THE STUDY DIRECTOR OTHER SCIENTISTS INVOLVED IN THIS STUDY STATEMENT OF QUALITY ASSURANCE UNIT SUMMARY RESUME 1. INTRODUCTION 2. MATERIALS AND METHODS 2.1 TEST SUBSTANCE 2.1.1 Identification 2.1.2 Formulation procedure 2.2 TEST SYSTEM 2.2.1 Animals 2.2.2 Environmental conditions 2.2.3 Pood and water 2.3 TREATMENT 2.3.1 Fasting of the animals 2.3.2 Administration of the test substance 2.3.3 Date of treatment and duration of the study 2.4 CLINICAL EXAMINATIONS 2.4.1 Clinical signs and mortality 2.4.2 Body weight 2.5 NECROPSY 2.6 DATA EVALUATION 2.7 PROTOCOLE ADHERENCE 2.8 ARCHIVING 3. RESULTS 3.1 CLINICAL EXAMINATIONS 3.1.1 Clinical signs (table 1) 3.1.2 Mortality (table 1) 3.1.3 Body weight (figures 1 and 2, tables 2 and 3) 3.2 PATHOLOGY (table 4) Does not contain TSCA CBJ ^ 4 4 8 8 8 9 9 9 9 10 10 10 10 10 10 10 11 11 _ 11 11 12 12 12 12 12 12 4. CONCLUSION 12 Figure 1: Body weight of treated rats (g) } 3 Figure 2: Body weight of C.I.T. historical control rats (g) 13 Table 1: Individual clinical signs and mortality 15 Table 2: Individual and mean body weight and weekly body weight change of treated 'rats(g) 16 Table 3: Mean body weight and weekly body weight change of C.I.T. historical control rats (g) 17 Table 4: Individual macroscopic examinations at necropsy 1 APPENDICES 1. Test article description 2. Diet formula 19 . 20 22 and 23 .-..atil&lfi 1 *>^r't-a' ^n/oiuu_y IMJ. luv-.J 1 ^-^ll^----ii--------liBil----1^11 /T-i-ULllcill .J.n.. STATEMENT OF THE STUDY DIRECTOR The study was performed in compliance with the principles of Good Laboratory Practice as described in: . O.E.C.D. principles of Good Laboratory Practice, Decision Concerning Mutual Acceptance of Data in the Assessment of Chemicals, C(81)30(fmal) Annex 2. May 12, 1981. . Decret'N0 90-206 du 7 mars 1990 concemant les Bonnes Pratiques de Laboratoire (Journal Officiel du 9 mars 1990), Ministere de 1'Industrie et de 1'Amenagement du Territoire. . Council Directive 87/18/E.E.C. of 18 December 1986 on the harmonization of laws, regulations or administrative provisions relating to the application of the Principles of Good Laboratory Practice and the verification of their applications for tests on chemical substances (O.J.nL 15 of 17.1.87). I declare that this report constitutes a true and faithful record of the procedures undertaken and the results obtained during the performance of the study. This study was performed at the Centre International de Toxicologie (C.LT.), Miserey, 27005 Evreux, France. Toxicology X.Manciaux Study Director Date: 14 May 1998 Doctor of Pharmacy OTHER SCIENTISTS INVOLVED IN THIS STUDY For Pharmacy: P. 0. Guillaumat Doctor of Pharmacy For Toxicology: C. Pelcot Study Supervisor ^pa^an^.Oo^.on.ainTSCAC. jJ.JL r't.'Lt-'^-'lA^UH tJ.i'-L. STATEMENT OF QUALITY ASSURANCE UNIT Type of inspections - Protocol Report Inspections 28 August 1997 11 May 1998 Dates Reported to Study Director (*) 2 September 1997 12 May 1998 Reported to Management (*) 2 September 1997 12 May 1998 At about the same time as the study described in this report, "process-based" and routine facility inspections of critical procedures relevant to this study type were made by the Quality Assurance Unit. The findings of these inspections were reported to the Study Director and to C.I.T. Management. The inspections were performed in compliance with C.I.T. Quality Assurance Unit procedures and the Good Laboratory Practice. The reported methods and procedures were found to describe those used and the results to constitute an accurate and complete reflection of the study raw data. P o ^ L. Valette-TaIbi Date: 14 May 1998 Doctor of Biochemistry Head of Quality Assurance Unit and Scientific Archives (*) The dates indicated correspond to the dates of signature of audit reports by Study Director and Management. 'Company 'Sanitised- Does Voi contain TSCA C" ^ii/oluay INO. iou4;) lAKj------MI----f^^11'^rocnem ^).A. 5 SUMMARY substance|mHHUUlHU^vas At the reques^^H^tocheinSA_JPai^^Defense, France, the acute oral toxicity of the test evaluated in rats according to O.B.C.D. (No. 401,24th February 1987) and E.C. (92/69/E.E.Q, Bi, 31st July 1992) guidelines. The study was conducted in compliance with the Principles of Good Laboratory Practice Regulations. Methods The test substance was administered by oral route to one group of ten fasted Sprague-Dawley rats (five males and five females). The test substance was administered in its original form, by gavage, at the dose of 2000 mg/kg, taking into consideration that its density was 0.848. Clinical signs, mortality and body weight gain were checked for a period of 14 days following the single administration of the test substance. All animals were subjected to necropsy. Results No death occurred at 2000 mg/kg. The general behaviour and body weight gain of the animals were not affected by treatment with the test substance. No apparent abnormalities were observed at necropsy. Conclusion IHBIHHIBRis Jnder_Qur experimental conditions, the oral LDo of the test substancy equal to or higher than 2000 mg/kg in rats. No signs of toxicity were observed at this dose. * According to the classification criteria laid down in Commission Directive 93/21/E.E.C. (27thApril 1993) adapting to technical progress for the eighteenth time Council Directive 67/548/E.E.C., the test substance does not present a significant acute toxic risk if swallowed. S^^OoesnoleontalnTSCACBI Company CH/Study JNo. 16U45 TAKSgBMBMHMBBnbIt Atochem S.A. 7 RESUME , A^ la demande de Elf Atochem 'S.A., Paris-la-Defense, France, la toxicite aigue du produit ^^BiBHIUBiHBHBBI^pres administration unique par voie orale chez Ie Rat a ete evanieeconformement aux lignes directrices de 1'O.C.D.E. (No. 401, 24 fevrier 1987) et de la C.E.E. (92/69/E.E.C., Bi, 31 juillet 1992). L'etude a ete realisee conformement aux regles de Bonnes Pratiques de Laboratoire. Methode Le produit a ete administre par voie orale a un groupe de 10 rats Sprague-Dawley (5 males et 5 femelles) mis ^ la diete hydrique. L'administration a ete effectuee avec le produit tel quel, par gavage, a la dose de 2000 mg/kg, en tenant compte de sa densite (d = 0,848). Les signes cliniques, la mortalite et revolution ponderale des animaux ont ete suivis pendant une periode de 14 jours apres 1'administration unique du produit. Un examen anatomopathologique a ete effectue sur tous les animaux. Resultats La mortalite est nulle a la dose de 2000 mg/kg. Aucun signe clinique n'est observe pendant 1'etude. L'evolution ponderale des animaux n'est pas influencee par le traitement. L'autopsie des animaux ne met en evidence aucune anomalie apparente. Conclusion Dans nos conditions experimentales, la DLO orale du produit^BBB^HUHw JBlBHBH--^st superieure ou egale a 2000 mg/kg chez le Rat. Aucun signe clinique n'est Selon les criteres de classification decrits dans la Directive 93/21/C.E.E (27 avril 1993) portant dix-huitieme adaptation au progres technique de la Directive 67/548/C.E.E., le produit ne presente pas de risque toxique aigu significatif en cas d'ingestion. .,,.--.---"1 U.T/^tudy INO. 101)43 iAKlJi^--------^--^[dr/\[Ocneni^.A. 6 1. INTRODUCTION The objective of this study was to evaluate the toxicity of the test substanc following a single oral administration in rats. In the assessment of the toxic characteristics of a test substance, determination of acute oral toxicity is an initial step. It provides information on health hazards likely to arise from a shortterm exposure by the oral route in Man. The study was conducted in compliance with: . O.E.C.D. guideline No. 401, 24th February 1987. . E.C. Directive No. 92/69/E.E.C., Bi, 31st July 1992. 2. MATERIALS AND METHODS 2.1 TEST SUBSTANCE 2.1.1 Identification____ The test substancepUUBHBIpsed in the study was supplied by Elf Atochem S.A. The test substance was identified as follows: . batch number: protocol and labellingg^:|^t|^H^^^^^^^k^^_. - . Elf Atochem filing number^--------------J _^ . description: ^ _.,,_ , . container: one plastic flask . date of receipt: 23 August 1997 ,. . storage conditions: at room temperature and protected from light . expiry date: August 1998. Data relating to the characterization of the test substance are documented in a test article description (presented in appendix 1) provided by the Sponsor. 'At the finalization of the study report, an analytical certificate was not available. Characterization of the test substance, which appropriately defines the tested batch, is under the responsibility of the Sponsor. 2.1.2 Formulation procedure The test substance was administered in its original form. ^ W ^ ^ ^tioesWteontalnTSCACW ,, 2.2 TEST SYSTEM 2.2.1 Animals Species, strain: rat, Sprague-Dawley ICO: OFA-SD (IOPS Caw). Reason for this choice: rodent species generally accepted by regulatory authorities for this type of study. Breeder: Iffa Credo, 69210 L'Arbresle, France. Number and sex: one group of ten animals (five males and five females). Age/weight: on the day of treatment, the animals were approximately 6 weeks old, and had a mean body weight standard deviation of 192 6 g for the males and 144 7 g for the females. Acclimatization: at least 5 days before the beginning of the study. Identification of the animals: the animals were identified individually by earmarks or eamotches. 2.2.2 Environmental conditions During the acclimatization period and throughout the study, the conditions in the animal room were set as follows: . temperature: 21 2C . relative humidity: 30 to 70% . light/dark cycle: 12 h/12 h . ventilation: approximately 12 cycles/hour of filtered, non-recycled air. The temperature and relative humidity were under continuous control and recording. The records were checked daily and retained. In addition to these daily checks, the housing conditions and corresponding instrumentation and equipment are verified and calibrated at regular intervals. The animals were housed in polycarbonate cages (48 cm x 27 cm x 20 cm). Each cage contained four to seven animals of the same sex during the acclimatization period and five rats of the same sex during the treatment period. Each cage contained dust-free sawdust (SICSA, 94142 Alfortville, France). Bacteriological and chemical analysis of the sawdust, including the detection of possible contaminants (pesticides, heavy metals), are performed regularly by external laboratories. The results of these analyses are archived at C.I.T. 2.2.3 Food and water All the animals had free access to A04 C pelleted diet (U.A.R., 91360 Villemoisson-sur-Orge, France), except as noted in "2.3.1 Fasting of the animals". Each batch of food was analysed by the supplier for composition and contaminant levels. The diet formula is presented in appendix 1/2. Drinking water filtered by a P.G. Millipore membrane (0.22 micron) was provided ad libitum. Bacteriological and chemical analysis of the water and diet, including the detection of possible contaminants (pesticides, heavy metals and nitrosamines), are performed regularly by external laboratories. The results of these analyses are archived at C.I.T. No contaminants are known to be present in the diet, drinking water or bedding material at levels which may be expected to interfere with or prejudice the outcome of the study. .^.<---la"TSCAC81 Cempany 2.3 TREATMENT 2.3.1 Fasting of the animals The animals were fasted for an overnight period of approximately 18 hours before dosing, but had free access to water. Food was given back approximately 4 hours after administration of the test substance. 2.3.2 Administration of the test substance As the test substance was anticipated to be non-toxic at 2000 mg/kg, a limit test was performed by administering 2000 mg/kg of the test substance to one group of ten animals (five males and five females). The test substance was administered in its original form taking into consideration that its density was 0.848 The administration was performed in a single dose by oral route using a stainless steel roundtipped probe (diameter: 18 G.I", Perfektum: Poffer & Sons Inc., New Hyde Park, New York 11040, U.S.A.) fitted to a 1 ml glass syringe (0.01 ml graduations. Record: Carrieri, 75005 Paris, France). The volume administered to each animal was adjusted according to body weight determined on the day of treatment. 2.3.3 Date of treatment and duration of the study The single administration was performed on 16 October 1997 in the morning (day 1) and was followed by a 14-day observation period until 30 October 1997 (day 15). 2.4 CLINICAL EXAMINATIONS 2.4.1 Clinical signs and mortality The animals were observed frequently during the hours following administration of the test substance, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day until day 15. Type, time of onset and duration of clinical signs were recorded for each animal individually. Time of death was recorded individually, in terms of the number of hours or days after dosing. 2.4.2 Body weight The animals were weighed individually just before administration of the test substance on day 1 and then on days 8 and 15. The body weight gain of the treated animals was compared to that of C.I.T. control animals with the same initial body weight. ^SaalM.Ooe^ot^ainTSCAC- 2.5 NECROPSY On day 15, all animals were killed by carbon dioxide asphyxiation and a macroscopic examination was performed. After opening the thoracic and abdominal cavities, a macroscopic examination of the main organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities) was performed. In case of macroscopic lesions, organ samples were taken and preserved in 10% buffered formalin. No microscopic examination was performed. 2.6 DATA EVALUATION Evaluation of the toxicity of the test substance following a single oral administration in rats should include the relationship, if any, between the animals' exposure to the test substance and the incidence and severity of all abnormalities including behavioural and clinical abnormalities, macroscopic lesions, body weight changes, mortality and any other toxic effects. 2.7 PROTOCOL ADHERENCE The study was performed in accordance with Study Protocol No. 16045 TAR and subsequent amendments, with the following deviation from the agreed Study Protocol: - the relative humidity recorded in the animal room was sometimes outside of the target ranges specified in the protocol. This minor deviation was not considered to compromise the validity or integrity of the study. 2.8 ARCHIVING The study documentation and specimens generated during the course of the study are archived at C.I.T., 27005 Miserey, Evreux, France, for 10 years after the end of the in vivo phase of the study. The archived study materials include: . protocol and possible amendments, . raw data, . correspondence, . final report and possible amendments. On completion of this period, the archived study materials will be returned to the Sponsor, or may be archived at C.I.T. for a further period. ,,^^^BoesBsc"a"'TSCACW Cil/^tudy JNO. 16U45 lAjbHHHHBHIHHIHbIt Atochem y.A. 12 3. RESULTS 3.1 CLINICAL EXAMINATIONS 3.1.1 Clinical signs (table 1) No clinical signs were observed during the study. 3.1.2 Mortality (table 1) No death occurred during the observation period. 3.1.3 Body weight (figures 1 and 2, tables 2 and 3) The body weight gain of the animals was not influenced by treatment. 3.2 PATHOLOGY (table 4) Macroscopic examination of the main organs of the animals revealed no apparent abnormalities. 4. CONCLUSION flUHHH^HFI8 Under our experimental conditions, the oral LDo of the test substance^dHBHH^IH equal to or higher than 2000 mg/kg in rats. No signs of toxicity were According to the classification criteria laid down in Commission Directive 93/21/E.E.C. (27th April 1993) adapting to technical progress for the eighteenth time Council Directive 67/548/E.E.C., the test substance does not present a significant acute toxic risk if swallowed. -^^^ ^ ^^oe CIT/Study No. 16045 TAFJJBIB------KS^11' Atoctiem S.A. i j Figure 1: Body weight of treated rats (g) Cprapany Sanitized. Does not contain TSCA CB? CIT/Study No. 16045 T^ ilf Atochem S.A. Figure 2: Body weight of C.I.T. historical control rats (g) -Male -Female e.g.: C.I.T. Historical data of animals dosed by the oral route: results of control animals from October 1995 to December 1997. ^es-nol eontain TSCACB1 L-ii/^tuay INO. iOU43 lA^nii----i------WjbifAtochem^.A. 15 Table 1: Individual clinical signs and mortality Dose Time Animals Mortality (mg/kg) Males Females 2000 30min 1 lh-2h-4h i-01-02-03-04-05 01-02-03-04-05 No D 2 to D 15 J mm: minutes h : hour D : day Clinical signs None ^-^' --- Table 2: Individual and mean body weight and weekly body weight change of treated rats (g) Dose ing/kg Volume Sex ml/kg Days Animals -------------------------------------------------------------------------- 1 (1) 8 (1) 15 2000 2.36 Male 01 199 69 268 55 323 02 197 75 272 64 336 03 184 67 251 50 301 04 187 64 251 37 288 05 193 67 260 60 320 M 192 68 260 53 314 SD 6 4 10 10 19 2000 2.36 Female 01 136 28 164 17 181 02 152 37 189 16 205 03 143 37 180 21 201 04 151 31 182 17 199 05 138 49 187 15 202 M 144 36 180 17 198 SD 7 8 10 2 10 (1) = Body weight gain M =Mean SD = Standard Deviation --SCl^CBl . ^oe^100^ -^^ed ^. Table 3: Mean body weight and weekly body weight change of C.I.T. historical control rats (g) BODY WEIGHT OF CONTROL RATS (g) mg/kg 0 ml/kg 10 Male Days 1 8 M 182 261 SD 10 12 n 45 45 0 10 Female M 147 190 SD 10 15 n 45 45 M mean SD standard deviation n number of animals 15 315 21 45 215 17 45 BODY WEIGHT CHANGE OF CONTROL RATS (g) T^C"lC*f=t mg/kg 0 \//"lll imf* ml/kg 10 QP'Y Male --..--.--...--.----.,--.--.--.-- 1 to Days -- --.--------.--.-------- 8 8 to 15 M 79 54 SD 7 13 0 10 Female M SD M : mean SD : standard deviation 43 25 8 ^ 7 e.g.: C.I.T. Historical data of animals dosed by the oral route: results of control animals from October 1995 to December 1997. company SanStted. Does not contain TSCA CBl Table 4: Individual macroscopic examinations at necropsy Dose mg/kg 2000 D:day Time D15 Males Animals Females 01-02-03-04-05 01-02-03-04-05 Macroscopic abnormalities None ^mfiW^^-^ . -^oeswt contain TSCACW APPENDICES 1. Test article description ^s^.--0"'8'"^' err/Study No. 16045 77 lit" Atochem S.A. TOXICOLOGY DEPARTMENT CONFIDENTIAL 11 August 1997 elfatochem s.a. La defense 10, Cours Michelet 92091 Paris-la-Defense cedex, France TEST ARTICLE DESCRIPTION IDENTITY Test article name CAS number EINECS number Purity Origin Batch Elf Atochem filing number ;RDATO, Serquigny PHYSICAL AND CHEMICAL PROPERTIES Appearance Specific gravity Boiling point Vapor pressure Flash point Solubility TOXICOLOGICAL INFORMATIONS AND USE SAFETY see fiche de donnees de securite STORAGE AND DISPOSAL Storage Expiry date Disposal in dark and at room temperature August 1998 incineration ganife0- DOSS ^sy>'.nts,,'" -- T"e-^"-''-"'? '-Sorop0^ 2. Diet formula IT Ref: A04 COMPLETE DIET RAT AND MOUSE MAINTENANCE DIET Appearance: 15 mm diameter pellets or powder Conditioning: 25 kg double paper bag with aluminium on the outside Daily portion: Rat 18-25 g. Mouse 5-10 g, water ad libitum. FORMULA % Cereals and cereal biproducts...... Vegetable protein (soya bean meal, yeast) .................................. Animal protein (fish).................... Vitamin and mineral mixture....... AVERAGE ANALYSIS % Calorific value (KCal/kg) Moisture........................... Proteins............................, Lipids................................ Carbohydrates (N.F.E.)..... Fibre.................................. Minerals (ash) ................... 2900 12 17 3 58.7 AMINO ACID VALUES (calculated in mg/kg) MB>^RALS (calculated in EGig/kg) Nat CMV val. val. Total P ,,, , Ca.,,,, K Na, , Ms. Mn Fe, Cu Zn,,,, Co...... I 5900 3300 6700 300 1900 50 90 15 40 T 0.3 0 5000 0 1600 100 40 150 15 45 1.5 0 5900 8300 6700 1900 2000 90 240 30 85 1.5 0.3 vnFAMINS (CEalculated perkg) Nat CMV val. val. Total Arginine ..... Cystine ....... Lysine......... Methionine. Tryptophan. Glycine....... FATTY ACID VALUES (calculated in mg/kg) Palmitic acid ..... Palmitoleic acid. Stearic acid......... Oleic acid........... Linoleic acid....... Linolenic acid..... 9800 2300 8500 3200 1900 8100 2600 Traces 500 8000 14500 Traces Vitamin A Vitamin D3 Vitamin B 1 Vitamin B2 Vitamin B3 Vitamin B6 Vitamin B 12 Vitamin E Vitamin K3 Vitamin PP Folic acid Biotin Choline Traces Traces 6mg 2mg 10 mg 1.3 mg 0.01 mg 15 mg 0.25 mg 60 mg 0.5 mg 0.04 mg 1200 mg 7500 IU 1500 IU Img 4.5 mg 6.5 mg 1.3 mg 0.01 mg 15 mg 2.25 mg 15 mg Omg Omg 400 mg 7500 IU 1500 IU 7mg 6.5 mg 16.5 mg 2.6 mg 0.02 mg 30 mg 2.5 mg 75 mg 0.5 mg 0.04 mg 1600 mg Available under quality "Control Ref:: A04 C " U.A.R., 7 rue Gallieni, 91360 Villemoisson - Tel: 01.69.04.03.57 - Fax : 01.69.04.81.97 (Ref. Doc. UAR: 1992) .Company ganged. Does not contain T8CA ^