Document 5DqO0OQdRxj740Vg5axqGw7dz

3) OECD 404-OPPTS 870.2500, Acute dermal irritation (rabbit), 132-004 -IC e' .I < Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 ACUTE DERMAL IRRITATION STUDY OF T-7485 APPLIED TO NEW ZEALAND WHITE RABBITS STUDY NUMBER: 132-004 SANITIZED SPONSOR 3M Corporate Toxicology 3M Center, Building 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3320 DEC 0 9 2003 SPONSOR'S REPRESENTATIVE John L. Butenhoff, Ph.D., DABT, CIH Senior Laboratory Manager Telephone: 651-733-1962 FAX: 651-733-1773 TESTING FACILITY Primedica Redfield Mail: P.O. Box 308 Deliveries: 100 East Boone St. Redfield, AR 72132 Telephone: 501-397-2540 FAX: 501-397-2002 STUDY DATES Study Initiation: May 22, 2000 Animal Phase Initiation: May 24, 2000 Animal Phase Completion: June 1, 2000 Study Completion: September 20, 2000 Page 1 1 ,\. Acute Dermal IrritationStudy in New Zealand White Rabbits Study Number: 132-004 GLP COMPLIANCE STATEMENT Study Number: 132-004 I certify that this study was performed in compliance with the United States Food and Drug Administration (FDA) Good Laboratory Practice Regulations (21 CFR Part 58) and OECD Regulations [C(81)30(final)] and that the report accurately reflects the raw data. Study Director Primedica Redfield Page 2 Acute Dermal irritation Study in New Zealand White Rabbits Study Number: 132-004 QUALITY ASSURANCE STATEMENT Study Number: 132-004 This study has been inspected and audited by the Quality Assurance Unit (QAU) as required by the Good Laboratory Practice (GLP) regulations promulgated by the U.S. Food and Drug Administration and OECD Regulations. The following is a record of the dates that audits/inspections were performed and reported by the QAU. I DATEOF AUD~T~~NSPECTION 05/18/00 TYPEOF AUDIT~~NSPECTION Protocol DATESREPORTETDO STUDY DIRECTOARND MANAGEMENT 05/18/00 05/24/00 Dose Administration, Dermal Scoring, and Clinical Observations I 06/02/00. 06/22/00 Amendment #I Raw Data, Formulations Raw Data, and Tables 05/24/00 06/02/00 06/22/00 06/29/00 I Draft Report Final Report 06/30/00 I 07119/00 I The report accurately reflects the original data. APPROVED BY: Quality Assurance Auditor Primedica Redfield Page 3 i Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 TITLE ACUTE DERMAL IRRITATION STUDY OF T-7485 APPLIED TO NEW ZEALAND WHITE RABBITS APPROVED BY: Study Director W Primedica Redfield Page 4 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 TABLE OF CONTENTS Page Cover Page.................................................. ................................................................................ 1 GLP Compliance Statement ............................................................................................................. 2 Quality Assurance Statement ........................................................................ ..................... 3 Title and Signature Page ......................................................................... 4 Table of Contents ............................................................................................................................. 5 Study Abstract ..... .................................................................. 6 0bjective............................ ............................................................. 7 Materials and Methods ................... ............................................................... 7 Archival Statement............................................................................................................................ 8 .......:.................................................................................. .... . 9 Conclusion ........................................................................................................................................ 9 Appendices .................................... ....................................................................................... 10 Appendix 1 - Individual Clinical Observations.......................................................................... 11 Appendix 2 - Individual Dermal Scores.................................................................................... 13 Appendix 3 - Individual Body Weights ..................................................................................... 15 Appendix 4 - Individual Body Weight Changes........................................................................ 17 Appendix 5 - Certificate of Analysis ......................................................................................... 19 Appendix 6 - Protocol and Amendment ................................................................................... 21 Appendix 7 - Protocol Deviations ............................................................................................. 33 Appendix 8 - Key Personnel....... .............................................................................. 35 Page 5 r Acule Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 ACUTE DERMAL IRRITATION STUDY OF T-7485 APPLIED TO NEW ZEALAND WHITE RABBITS STUDY ABSTRACT The objective of this study was to determine the acute effect(s) of a single dermal application of test material to New Zealand White rabbits with a fourteen-day recovery. The animals were acclimated for 16 days prior to Study Day 1 and were examined by the Staff Veterinarian prior to being released for use on the study. During the acclimation period, the animals were habituated to Elizabethan collars three times, increasing the time period to at least six hours. Three females were assigned to the study. The study design was as follows: Group Number Group Designation I Number of Animals Females 1 Limit Dose 500 3 On Study Day 1, the test material was applied to a small area (approximately 6 cm') of skin and covered with a gauze patch, which was held in place with non-irritating tape. Elizabethan collars were placed on the animals during the exposure period of approximately four hours. At the end of the exposure period, residual test material was removed with warm tap water and gauze. Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). On Study Day 1, clinical observations were recorded predose and approximately hourly for four hours postdose. Thereafter, clinical observations were recorded once daily. During the nine-day study, no adverse'findings were reported, with the exception of one rabbit exhibiting decreased appetite on Study Days 2 to 4. Scoring of the dose site was performed at the time of patch removal and at approximately 24, 48, and 72 hours after patch removal using the Draize scoring system. As indicated in all rabbits, 500 mg of T-7485 failed to induce erythema, edema, or other possible dermal findings during the scoring periods. Body weights recorded on Study Days 1 and 8 indicated a positive weight gain for Study Days 1 through 8 of 0.41, 0.25, and 0.19 kilograms for rabbits #1652, #1654, and #1656, respectively. On Study Day 9, rabbit #I652 lost 30 grams of body weight when compared to Study Day 8. In contrast, on Study Day 9 rabbit #I654 gained 30 grams of body weight while rabbit #I656 weight was unchanged when compared to Study Day 8. Due to lack of additional animals, which would be needed for meaningful statistical analyses, these data suggest the weight changes are within normal body weight variability and not related to application of test material. In conclusion, a single exposure of 500 mg of T-7485 to rabbits failed to induce erythema, edema, or other possible dermal findings during the scoring periods. Page 6 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 OBJECTIVE The objective of this study was to determine the acute effect(s) of a single dermal application of test material to New Zealand White rabbits with a fourteen-day recovery. MATERIALS AND METHODS TEST AND CONTROL MATERIALS: The test material and vehicle were identified as follows: Test Material: Lot Number: Date Received: Physical Description: Storage: Amount Received: T-7485 (Potassium Perfluorobutane Sulfonate, PFBS) 2 April 6, 2000 White Powder Room Temperature 500 grams Vehicle: Supplier: Lot Number: Carboxymethylcellulose (medium viscosity) Sigma Chemical Co. 69H0028 The Sponsor assumed responsibility for characterization (identity, purity, and stability) determinations of the test material. The test material was inventoried when received at Primedica Redfield, and a record of all test material usage was maintained. The Certificate of Analysis for the test material is in Appendix 5. Doses were prepared by Primedica Redfield on the day of use. The vehicle was prepared to a concentration of 1% by mixing 10 grams of powdered medium viscosity carboxymethylcellulose with deionized water using a Waring blender. Additional deionized water was added to yield 1000 mL of prepared vehicle. The vehicle was stored refrigerated when not in use. The test material was moistened with the vehicle prior to administration to the animals. TEST ANIMALS: Healthy female New Zealand White rabbits were received from Myrtle's Rabbitry, Inc. for use on the study. The animals were individually housed in stainless steel cages. Animal identification consisted of uniquely numbered ear markings and cage cards. On Study Day 1, the animals were approximately 14 weeks old and weighed 2.78 to 3.25 kilograms. Teklad Certified Rabbit Diet #8630 was provided at approximately 125 grams per day, and filtered tap water was provided ad libitum. The feed and water were routinely analyzed for contaminants. There were no known contaminants in the feed or water that would be expected to affect the results of the study. The results of these analyses are on file at Primedica Redfield. Environmental controls were set to maintain temperatures of 16" to 22C (61" to 72F) with a relative humidity of 30% to 70%. These parameters were recorded at least once daily. A 12:12 hour lightdark cycle and ten or greater air changes per hour were maintained in the animal room. GROUP DESIGNATIONAND TREATMENT: The animals were acclimated for 16 days prior to Study Day 1 and were examined by the Staff Veterinarian prior to being released for use on the study. During the acclimation period, the animals were habituated to Elizabethan collars three times, increasing the time period to at least six hours. Three females were assigned to the study. The study design was as follows: Page 7 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Group Number Group Designation Dosage Level (mg) Number of Animals Females I 1 1 7- 1 ~ ~ 1 Limit Dose ~ 500 3 Hair was removed by clipping the dorsal area of the trunk of all animals at least one hour prior to test material administration. The dose site and an area of adjacent untreated skin of similar dimensions were marked at the corners with an indelible marker. On Study Day 1, the test material was applied to a small area (approximately 6 cm2)of skin and covered with a gauze patch, which was held in place with non-irritating tape. Elizabethan collars were placed on the animals during the exposure period of approximately four hours. At the end of the exposure period, residual test material was removed with warm tap water and gauze. JUSTIFICATION FOR SPECIES SELECTION, ROUTE OF ADMINISTRATION, AND DOSE LEVEL: Rabbits are an animaf model for acute toxicity studies of this type. The number of animals assigned to the study represented the minimum required to meet the objectives of the study and the OECD guidelines (#404). In the assessment and evaluation of the toxic characteristics of a substance, determination of acute dermal toxicity is useful where exposure by the dermal route is likely. Dose levels were chosen in accordance with OECD guideline M04. CLINICAL OBSERVATIONS: Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). On Study Day 1, clinical observations were recorded predose and approximately hourly for four hours postdose. Thereafter, clinical observations were recorded once daily. DERMAL SCORING: Scoring of the dose site was performed at the time of patch removal and at approximately 24, 48, and 72 hours after patch removal using the Draize scoring system found in the protocol (Appendix 6). BODY WEIGHTS: Body weights were recorded pretest and on Study Days 1, 8, and 9. The pretest body weights are not included in this report but are located in the raw data. STUDY TERMINATION: On Study Day 9, the study was terminated. All animals were humanely euthanized and discarded. No necropsies were performed. STATISTICS: No statistical analyses were performed ARCHIVAL STATEMENT All original data and the original final report will be retained at Primedica Redfield's archives, located at 100 East Boone Street, Redfield, Arkansas, for a period of five years after issuance of the final report. After this time, the Sponsor will be contacted for disposition instructions. Page 8 Acute Dermal IrritationStudy in New Zealand White Rabbits Study Number: 132-004 RESULTS CLINICAL OBSERVATIONS: Observations for mortality and moribundity were recorded twice daily (a.m. and p.m.). On Study Day 1, clinical observations were recorded predose and approximately hourly for four hours postdose. Thereafter, clinical observations were recorded once daily. During the nine-day study, no adverse findings were reported with the exception of one rabbit exhibiting decreased appetite on Study Days 2 to 4 (Appendix 1). DERMAL SCORING: Scoring of the dose site was performed at the time of patch removal and at approximately 24, 48, and 72 hours after patch removal using the Draize scoring system. As indicated in all rabbits, 500 mg of T-7485 failed to induce erythema, edema, or other possible dermal findings during the scoring periods (Appendix 2). BODY WEIGHTS AND BODY WEIGHT CHANGES: Body weights recorded on Study Days 1 and 8 indicate a positive weight gain between Study Days 1 through 8 of 0.41, 0.25, and 0.19 kilograms for rabbits #1652, #1654, and #1656, respectively (Appendix 4). On Study Day 9, rabbit #I652 lost 30 grams of body weight when compared to Study Day 8 (Appendix 3). In contrast, on Study Day 9 rabbit #I654 gained 30 grams of body weight while rabbit #I656 weight was unchanged when compared to Study Day 8 (Appendix 3). Due to lack of additional animals, which would be needed for meaningful statistical analyses, these data suggest the weight changes are within normal body weight variability and not related to application of test material. CONCLUSION In conclusion, a single exposure of 500 mg of T-7485 to rabbits failed to induce erythema, edema, or other possible dermal findings during the scoring periods. Page 9 Acute Dermal IrritationStudy in New Zealand White Rabbits Study Number: 132-004 APPENDICES Page 10 Acute Dermal IrritationStudy in New Zealand White Rabbits Study Number: 132-004 APPENDIX 1 INDIVIDUAL CLINICAL OBSERVATIONS Page 11 PROTOCOL 1 3 2 - 0 0 4 : ACUTE DERMAL IRRITATION STUDY OF T - 7 4 8 5 APPLIED TO NEW ZEALAND UHITE RABBITS A_PPE_NDI_X 1_: C_LIN_ICA_L O_BSER_VAT_ION_S -_IND_IVID_UAL_- _FEM_ALE_RAB_BIT_S ____________________---.----------------- D.O. .S.E. .G.R.W. .P. .1. . . . . . . . . . . . . . .5.0.0. .M.G. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . R-A_BBIT_# _ _ _ _ _ _DES_CRI_PTIO_N _ _ _ _ _ _ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 1652 SD( 2- 4 ) DECREASED APPETITE 1654 NO ADVERSE FINDINGS 1. .6.5.6. . . . . . . . . . . . . . . . . . . . . . .N.O. .A.D.V.E.R.S.E. .F.I.N.D.I.N.G.S. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . SD = STUDY DAY A n i m a l s with NO ADVERSE FINDINGS w e r e n o r m a l u n t i l their scheduled t e r m i n a t i o n . 9(D (D 2 N ~~ Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 APPENDIX 2 INDIVIDUAL DERMAL SCORES Page 13 STUDY NUMBER 132-004 APPENDIX 2 INDIVIDUAL DERMAL SCORES Acute Dermal IrritationStudy in New Zealand White Rabbits Study Number: 132-004 APPENDIX 3 INDIVIDUAL BODY WEIGHTS Page 15 EE s---------- i moran 52:04: ACUTEDAWA ARIAT STRYOF 1-743 OPEDTOMEVHAAR ITE ITE i 7 ii it & Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 APPENDIX 4 INDIVIDUAL BODY WEIGHT CHANGES Page 17 En e w EE --FE ------ ii photoco, 182-004: ACUTE BERNAL IANITAT IGN STAYOF 1-TU8S APPLIESTOKEW HALAW WACTE BABNITS k i i : i i ' i Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 APPENDIX 5 CERTIFICATE OF ANALYSIS Page 19 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Certificate Of Analysis C4F9S03K+(PFBS), Lot 2 March 10,2000 This sample was analyzed using , and elemental analyses techniques. The results of these tests show the sample to contain the following weight percent composition: 0.13 % c4F9so3x+ 99.82 % 0.04 % Additionally,the isomer distribution of the sample was determinedusing ' contain the followingweight percent composition: .c techniques and found to I CF,(CF2b-SOi K' (Normal chain) 97.86 % 1.96 Yo Page 20 ~~~ ~ ~ Acute Dermal irritation Study in New Zealand White Rabbits Study Number: 132-004 APPENDIX 6 PROTOCOL AND AMENDMENT Page 21 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Promo1 Number: 132434 ACUTE DERMAL IRRITATION STUDY OF T-7485 APPLIED TO NEW ZEALAND WHITE RABBITS PROTOCOL NO.: 132-004 OBJECTIVE: Determine the acute effect(s) of a single dermal application of test material to New Zedand White rabbits with a fourteenday recovery. LOCATION OF STUDY AND CONDITIONS OF TESTING: It is the intention of 3M Pharmaceuticals,through the conduct of this study, to generate animal safely data that may be submitted to regulatory authorities. Primedica Redfield, 100 East Boone Street, R-, Arkansas, 72132, is accredited by AAAlAC and licensed by the United States Department of Agricultwe to conduct research in laboratory animals. All the conditions of testing will conform to the Animal wdfrre Act (CFR 9) and its amendments. Primedica Redfield will follow all requirements specified in this approved protocol and all applicable governmental regulations regarding Good Laboratory Practices as well as Primedica Redfield's Standard Operating Procedures. Changes in the protocol may be rnade by consultation with and approval from 3M Pharmaceuticals followed by written verification of the change. 3M Pharmaceuticals reserves the right to inspect facilities and procedures used for this study by meam of announced or unannounced site visits. Primedica Redfieldwill n o m 3M Pharmaceuticalsp. rom.ptly_by_ telephone prior to release of any data for review. SPONSOR 3M 3M Center, Building 220-2E-02 P.0. Box 33220 St Paul, MN 55133-3320 SPONSOR'S REPRESENTATIVE: John L. Butenhoff, Ph.D., DABT, CIH Senior Laboratory Manager Telephone: 651- 73344 ba FAX: 651-733-1773 m,$,/@ TESTING F A C I L m Primedica Redfield Mail: P.O. Box 308 Deliveries: 100 East Boone St Redfield, AR 72132 Telephone: 501-397-2540 FAX: 501-397-2002 PERSONNEL: Study Director: John Seng, Ph.D. Principle Investigator: Karen Tranter. B.A.. LATG Veterinarian: Allan Manus, D.V.M., M.Sc.. ACL4M Head Technician: Catherine, Morris, A U T Pathologist: TBD PROPOSED STUDY DATES: Experimental Start Date: May 24,2000 Experimental Termination Date: June 7,2000 Draft Report Date: July 18, 2000 Page J of 9 5/23/00 Page 22 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Prolocol Nwnber: 132-004 1. REGU~ITORCYOMPLIANCEAND QUALIWASSURANCE This study will be conducted in accordance with the following Good Laboratory Practice RegulationslStandardslGuidelines: 0 21 CFR58 0 C(81)30 (Final) (OECD) The Quality Assurance Unit, in accordance with Primedica Redfields Standard Operating Procedures (SOPS),will audit the protocol, study conduct, and the final report. 2. INSllNTlONAL ANIMALCARE AND USE C O M M I T N REVIEW The protocol will be reviewed and approved by Primedica Redfields InstitutionalAnimal Care and Use Committee (IACUC) for compliance with regulations prior to study initiation. In the opinion of the Sponsor, indicated by the signature on this protocol, the study does not unnecessarily duplicate any previous work 3. MATERIALS TESTMATERIAL /DEMlFlCA77ON T-7485 (Potassium perfluorobutane sulfonate, PFBS) The lot number will be listed in the raw data and final report PHYSIW\LDEsCRlPnoN White powder VEHICLE Carboxymethylcellulose(aqueous, medium viscosity) IDENl7FlCATION The lot number will be listed in the raw data and final report ' PHYSICXDLESCRIPTIOWNhite powder STORAGE CONDITIONS The bulk vehicle and test material will be stored in its original container at room temperature. ANALYTICAL CHEMISTRV The Sponsor assumes responsibility for characterization (identity, purity. and stability) of the bulk test material (including under test conditions). Information on the composition and method of synthesis of the bulk test material will be held by the Sponsor. The doses, unless otherwise stated, will be calculated assuming the test material to be 100% pure. DOSE PREPAR4TlONAND CONCENTRATION Doses will be prepared by Primedica Redfield on the day of use. The carboxymethylcellulosewill be preparedat 1%in deionized water and may be prepared prior to Study Day 1. Page 2 of 9 5R3i00 Page 23 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Protocol Number: 132-004 INVENTORY The test material container@)will be inventoriedwhen received at the testing laboratory, and a record of all test material use will be maintained. The technician dosing the animals will: 1) Record the weight or volume of the test material container immediately before and after dosing; 2) Compute and record the amount used and compare it to the theoretical amount immediately after dosing; 3) Keep the formulated test material use log in the animal study books at all times; 4) Assure the dose used is within *IO% of theoretical. TEST MATERIARLETENTION Unusedtest material may be returned to the Sponsor or designeeat !he terminationof the study, or retained for use on future studies. The Sponsor will be notified in advance of shipping, and a transmittal letter will accompany the shipment. The material will be packed in a suitable container to maintain the conditions specified by the Sponsor during transit plus an adequate margin of safety for any transit delays. SAFETYPRECAUT~ONS General safety precautions as required by Primedica Redfiekl's policies and procedureswill be followed. The Sponsots Representativewill be notified of any personnel exposures requiring a physician's examination or care. 4. ANIMALS Species: StraidSource: Age at Initiation: Weight at Initiation: Number and Sex: Identification: Rabbit New Zealand White, Myrtle's Rabbitry, Thompson Station, Tennessee Approximately 12 weeks Approximately 2.0 to 3.5 kg at initiation Three female (female nulliparous and non pregnant) Indelible ink and colorcoded cage card ANIMALHUSBANDRY HOUSING The animals will be individually housed in stainless steel cages. The cages conform to standards set forth in the Guide for the Care and Use of Laboratow Animals, National Academy Press, Washington, D.C., 1996. FEED Teklad Certified Rabbit Diet #I3630 will be provided at approximately 125 grams per day. This diet is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. Results of the manufacturets analyses are on file at Primedica Redfield. WATER Filtered tap water will be provided ad libitum. Samples of the water are analyzed for total dissolved solids. hardness, and specified microbiological content, and for environmental contaminants. Results of these analyses are on file at Primedica Redfield. Page 3 of 9 5119/00 Page 24 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Protocol Number: 132-Ow CONTAMINANTS There are no known contaminants in the feed or water that would be expected to interfere with this study. ACWMATION Environmentalcontrols will be set to maintain a temperature of 16"to 22C (61"to 72F)with a relative humidity of 30% to 70%. These parameters will be recorded at least once daily. A 1212 hour lightdark cycle will be maintained. Animals will be acclimated for a minimum of seven days prior to the study start. An adequate number of extra animals will be purchased so that no animal in obviously poor health is placed on test. At the request of the Study Director, excess animals may be used as replacement animals. Animals will be examined by the Staff Veterinarian prior to release of that shipment of animals for use on the study. During pretest, the animals will be habituated to the collars used during the study at least three times, increasing the minimum time period of at least six hours. Group Number 1 Group Designation Limit Dose Dosage Level (mol 500 Number of Animals Females 3 Page 4 of 9 Page 25 511 9/00 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Proloco1 Number: 132-Ow JUSTIFICATION FOR ROUTE OF ADMINISTRATIOANND DOSELEVEL In the assessment and evaluation of the toxic characteristics of a substance, determination of acute dermal toxicity is useful where exposure by the dermal route is likely. Dose levels were chosen by the OECD guideline M04. ANTEMORTEM OBSERVATIONS CLINICAL OBSERVATIONS Observations will be performed and recorded twice daily (a.m. and p.m.) for moribundity and mortality. Clinical observations will be recorded at least once each day. Observations will be recorded predose and approximately hourly for four hours postdose on the first day of dosing then daily thereafter for at least 14 days. Observations should includechanges in the skin and fur,eyes, and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, and somatomotor activity and behavior pattern. Particular attention should be directed to observation of tremors, convulsions, salivation, diarrhea (liquid feces), lethargy, and coma. DERMALSCORING The scoring will be conducted approximately60 minutes, and then at 24, 48, and 72 hours after patch removal using the Draize scoring system in Attachment A (Draize et. a/. 1944,J. Phanacol. Exptl. Them. 82:377390). BODY WEIGHTS Body weighk will be recorded pretest. on Study Day 1, approximately weekly, and at termination. POSTMORTEOMB S E R V A ~ O N S MORIBUNADNIMALSAND ANIMALSFOUNDDEAD Animals unlikely to survive until the next scheduled observation will be weighed, euthanized, and necropsied. Animals found dead will be weighed and necropsied. In either case, blood samples will not be collected and a complete necropsy will be performed as detailed below. All animals surviving to the end of the study may be returned to the testing facility's methodology population. NECROPSY All animals that die during the study will be subjected to a complete necropsy examination. A complete necropsy is defined as examination of the external surface of the body, all orifices, and the cranial, thoracic, and abdominal cavities and their contents. Gross lesions will be preserved in a suitable fixative for possible histopathology; all other tissues will be discarded. Histopathology will be provided at additional expense to the Sponsor. Page 5 of 9 5/22/00 Page 26 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Prorocol Number: 132-004 7. REPORT An audited draft report will be sent to the Sponsor. Revisionsto the initial draft report will be provided to the Sponsor by mail or fax transmission as printed copies of the corrected pages only. Additional revisions or complete copies of the revised draft reports will be provided at additional expense to the Sponsor. Three copies (one bound, two unbound) of the approved find report will be submitted to the Sponsor approximately two weeks after approval of the draft report The final report will include all elements requiredlrecornmendedby the regulations and guidelines. ..The report will include, but is not limited to, those items listedbelow: Descriptive text of the study objective Summary Test material identification (Certificate of Analysis, stability data, and analytical analysis if any are performed) Methods Results and conclusions Body weights Body weight changes Dermal irritation scoring A copy of the protocol, all protocol amendments, and all protocol deviations that may affect the integrity of the study 8. MAINTENANCOEF RAW DATAAND RECORDS Original data or copies thereof, will be available at Primedica Redfield to facilitate auditing the study during its progress and before acceptance of the final report. When the final report is completed, all original paper data and the original final report will be retained in the archives of the laboratory for at least five years. Wet tissues, slides, and blocks (if generated) will be retained at Primedica Redfield for one year. After one year, the storage of the wet tissues, slides, and blockswill be negotiated with the Sponsor. After five years, the storage of the paper data will be negotiatedwith the Sponsor. The Sponsor will be notified prior to disposal of any original study data. 9. QIJALITYASSURANRCEEVIEW This is to certify that this protocol has been reviewed by Quality Assuance. - ---- Fran Pattillo. M.S.. RQAP-GLP Manager of Quality Assurance Primedica Redfield 4 - 5 3 - CD Date Page 6 of 9 5/22/00 Page 27 ~~~~ ~~~ Acute Dermal irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol 10. APPROVALS " John L. Butenhoff, Ph.D, DABT, CIH Date Senior Laboratory Manager J d n Seng. Ph.D. Date Study Director Primedica Redfield Protocol Nuder: 132-004 Page 7of 9 Page 28 511W O O Acute Dermal IrritationStudy in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Pro1ocol Number: 132-Ow ATTACHMENT A DERMAL IRRITATION SCORING SYSTEM Page 8 of 9 Page 29 511 9/00 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Proiocol Number: 1 3 2 W ATTACHMENT A DERMAL IRRITATION SCORING SYSTEM (from Drake, 1944) ERYTHEMA No Erythema .... ............................................... Very Slight Erythema (barely perceptible) .................................... Well Defined Erythema ................................................................... Moderate to Severe Erythema ...................................................... ............................. Severe Erythema (beet red) to Slight Eschar Formation(injuries in depth) ......................... VALVE 0 1 2 3 4 EDEMA VALUE No Edema ........................................................................ ..................................... 0 Very Slight Edema (barely perceptible) ................................................................. 1 Slight Edema (edges of area well defined by definite raising) ...... .............................. 2 Moderate Edema (raised approximately 1 mrn)..................................................................... 3 Severe Edema (raised more than 1 rnm and extending beyond the area of exposure) ....... 4 Other dermal effects: Atonia Blanching Desquamation (Epidermal scaling/flaking) Fissuring Necrosis Scab Thickening Ulceration Page 9 of 9 511 9/00 Page 30 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Amendment Protocol Number: 132-004 STUDY NUMBER: AMENDMENT: STUDY TITLE: DATE ISSUED: 132-004 1 ACUTE DERMAL IRRITATION STUDY OF T-7485 APPLIED TO NEW ZEALAND WHITE RABBITS June 9,2000 AMENDMENT # 1 ITEM 1: CHANGED FROM: CHANGED TO: REASON: Protocol 3M Pharmaceuticals 3M Corporate Toxicology Sponsor requested clarification. ITEM 2 CHANGED FROM: CHANGED TO: REASON: Page 1, SPONSOR'S REPRESENTATIVE Telephone: 651-737-2678 Telephone: 651-733-1962 Correction of telephone number. ITEM 3: CHANGED FROM: CHANGED TO: REASON: Page 1, Proposed Study Dates: Experimental Termination Date: June 7, 2000 Experimental Termination Date: June 1, 2000 Study terminated early, no abnormalities seen. ITEM 4: CHANGED FROM: CHANGED TO: REASON: Page 5, Section 5, Clinical Observations Observations will be recorded predose and approximately hourly for four hours postdose on the first day of dosing then daily thereafter for at least 14 days. Observations will be recorded predose and approximately hourly for four hours posidose on the first day of dosing then daily thereafter for up to 14 days. Study terminated early, no abnormalities seen. 1 Page 31 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 Primedica Redfield Protocol Amendment Protocol Number: 132-004 STUDY NUMBER: AMENDMENT STUDY TITLE: DATE ISSUED: 132-004 1 ACUTE DERMAL IRRITATION STUDY OF T-7485 APPLIED TO NEW ZEALAND WHITE RABBITS June 9,2000 AMENDMENT # 1 APPROVALS: John L. Butenhoff, Ph.D., DABT, CIH Senior Laboratory Manager 3M Corporate Toxicology Date J n Seng, ph.D. S t d y Director Primedica Redfield Date 2 Page 32 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 APPENDIX 7 PROTOCOL DEVIATIONS Page 33 Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 PROTOCOL DEVIATIONS There were no protocol deviations that adversely affected the integrity of the study. Page 34 ~~ ~ ~ Acute Dermal Irritation Study in New Zealand White Rabbits Study Number: 132-004 APPENDIX 8 KEY PERSONNEL Page 35 Acute Dermal irritation Study in New Zealand White Rabbits Study Number: 132-004 KEY PERSONNEL Study Number: 132-004 Study Director: ............................................... Study Coordinator: ......................................... Head Technician ............................................ Veterinarian: .................................................. Quality Assurance: ......................................... Report Coordination: ..................................... Report Preparation: ....................................... John Seng, Ph.D. Karen Tranter, B.A., LATG Catherine Morris, ALAT Allan Manus, D.V.M., M.Sc., ACLAM Shevonya Noble, B.A. Amy Babb, B.S. Tarnmy Peckharn, Ad. Asst. Tarnrnie Hughey, Ad. Asst. Page 36