Tobacco Documents Online

- U7JU15 N-1 i1;~)4 -(7-iN LLU.i JUJdOL 1Pa1rU nVel.u INBIFO Institut fur bioiogische Forschung GmbH ,-•-+ x~rs iar.rr,i.®r.m es 1~1'~ Gontract Research 1! Taloplione (0 22 Oa) 303-7 i, Telefax (0 22 03) 303-362 xvwr-0 tr,ariuc tCcr bldagische Fo~ TELEFAX ~ Dr. GeorSe Patskan O Philip Marrts U.S.A Research Center. Richmond, Va. 23261 1 CTt ~ Fax-No.: 001-804-274--28 ~ YOUr RPt Your Letter Our Ref. wREJBVI GF:a o=s.nac Cologne, 7-Jun-s404 ~ Re_, Comments to Draft "Recommended protocols for the evaluation of biolopica1 activity of low-ignitlon cigaraftas" Battelle No Lhwest, BNW No. 22591, May 16, 1994 Dear George: Please firtd attached the critical comments of prs. R6mer, Hau(3mann, and Walk to the BNW s LIP protocol: Certeral remarks- . I The recommendation, Wt to compare the chronic effects of sidestres.m cigarette smoke from an LIP to a reference cigarette because of probable Insensitivity of the available animal modes (page 22) is not understandable. The limited accuracy of the applied bioassays is an important issue when evaluating the overall performance of a given LIP ugarette type. Therefnre this issue must be addressed and supported by appropriate data. To I. intrQduction no comments To it. Qenotoxig~%dies The authors propose to use five Salmonella strains, although the CPSC Toxicity Testing Plan states that "two strains ... (TA9$ and TA10Q) provide an adequate evaluation of condensate mutagenicity". Trade Reg'kter. Q4logne NRS 307 Genetal Mandge': Gr. Wdf Re>nhghaus 28/33

07/06 '94 11:54 '8`4A 2203 303362 INBIFO Koeln 444 RCA Z001 INBIFO Institut fiir bioloqische Forschung GmbH Cantract Resean:h Talophone {0 22 03} 303-i Telefax (0 22 03) 303-362 >ti'$M lrmtitrt fitr hidoa&hO Fon3dklQ 41ntbH • Fu4gws:ra6a 3• 0•511a9 C0loqne TELEFAX Dr. George Patskan Ref. YourLetter Philip Nlorris U.S A W REJBVI . Our Ref, Research Center. GPA_LF3.C3dG Richmond, Va. 2:3261 t`ax-Nc,: QQ1-804-274•2$91 Cologne, 7-sun-9494 Re_- Comments to Draft "Recor»mendsd protocols for the evaluation of biologieal activity of low-ignition cigaraftas" BatteiDe Northwest, BNW No. 22591, May 16, 1994 Dear George: Please find attached the critical camments of Drs. Ramer, Hau(3mann, and Walk to the SNW's LIP protocol: Qgnerai re.mrks:. The recommendation, =t to compare the chronic effects of aiciestrea.rtti cigarette smoke from an LIP to a reference cigarette because of probable insensitivity of the available animal modes (page 22) is riot understandable. The limited accuracy of the applied bloassays is an important issue when evaluating the overall performance of a given LIP cigarette type. Therefore this issue must be addressed and supported by appropriate data, To !, introduction no comments To 11. Genotoxid,ly ftdies The authors propose to use five Salmonella strains, although the CPSC Toxicity Testing Plan states that "two strains ... (TA98 and TA1d0) provide an adequate evaiuation of condensate mutagenicity". Trade Rogiate, : Cologne HPa 3-87 V183 Generdl Mandgar: br. Wdf RelnVghaus

07/06 '94 11:55 V49 2203 303382 INBIFO Koe1n f INB11=0 insGtut fur bioiogische Forschung GmbH • KOIn - un.-, a~ to I2z. . Patskan aaa RCA There is no information on the statistical analyses to be performed, especially not on the criteria which wili be used to discriminate two types of cigarettes. The description of he test procedures are too generic, a lot of details are missing. A detailed protocol is needed in compliance with GLP. It appears that the authors have no special experience with cigarette condensate, which is essential to get meaningful results in the Ames assay. The authors are well aware that the results received in the Mammalian Cell Transformation Assay "are often scatterec!" (which is an understatement), but they do not comment on the fact, that up to now that assay has not shown its ability to give quantitative results. It should be clearly stated that there Is no Information In the literature about the usefulness of this assay to discriminate different cigarette types. As for the Ames Assay, important information on the proposed evaluation is miss;ng. To iil. Whqi!2 Animal Bioassav A. GeneralQomments It is reasonable to use only one species for the subchronic and chronic in9^aiation assays. However, the reference to the rat in this chapter is to early as compared to the thorough discussion of relevant species which follows. B. Inhalation Bioassay Is it possible to further substantiate the notion that, for the rat, the nicotine rather than the carbon monoxide concentration would be the limiting exposure factor? This statement contradicts our experience. With regard to the susceptibility of the various species to cigarette smoke-induced respiratory tract carcinogenesis it is conceded that neither of the species discussed showed clear responses in the past - with two exceptions. the dog model which is to lavish, and the BlO hamster. However, it should be mentioned that the latter does not seem to be available any more. A longer exposures period of 16 hours/day Is discussed which is said to be adapted to human smoking behavior. However, for the suggested studies only 1 hourtday is planned - without any comment on the deviation. There is not much of an discussion of the rat strain recommended, e.g., with regard to longevity or spontaneous tumor rate in the respiratory tract, 2002 N ~ CIA C ~ ~ N C1t N U1 iG9a

1 07!08 ' 94 11 ; 55 'js•f9 2203 303302 IABIFd Koeln --+ RCA s 1Nt31FO Institut for biologische Forschunq GmbH • K6In - un-94 sx to Dr. Cr. stskan Pagc 3 C. Skin Carcinacaenicity The proposed protocol is almost identical to that which would be proposed by us for this bioassay for this purpo5e. This includes, e,g., mouse strain, housing and application, group size, application volume, frequency and vehicle, tar quality control, In-life observations, definition of macroscopic tumors. Recommended protocoi madifications;. • 1Ne propose an initial dose adaptation period of several weeks to allow the mice to adapt to the ao<te toxicity of the high doses of tar. This wouid result in almost no loss of mice during the initial phase of the application period and would prevent the expensive 10% larger group size proposed by Batteile. • We propose to apply the weekly dose of 125 mg/mouse on 5 sucoessive days (2i; mg/mouse x day) instead of 3 spaced applications of 40 mg/mouse x day to further reduce acute toxicity. To iV. Estimated osts The estimrxted costs are approximately 3011A higher than the usual market prices (BNW are usually ccharging above average costs). Appendices A. and B. The cigarette conditiQning process does not adhere to the specific (S0 requirements, Further parameters for smoke generation should be checked, e.g., puff volume and puff count, to prove constant smoking conditions and adherence to ISO conditions. Detailed clinical observations and body weight determinations should be performed weekly rather then biweekly in aI 3-week inhalation study. The use of carbon dioxide anesthesia to kill tne rats is questioned due to expected haemcrrhage. Renne et al. as cited for the larynx sections do not investigate the iaryngotracheal junction at which cigarette smoke-induced effects may also be expected at the concentrations planned. In addition, this citation seems to be wrong. No details are given for the statistical evaluation of mortality and tumor Incidence data. Best regards, 2003 ,0M