TABLE OF CONTENTS TAB TITLE PRESENTER i Introduction Rick Solana 2 Toxicology Testing Approach George Patskan 3 Surrogate Markers Suitable for Testing Hans-Juergen Haussmann 4 Lessons Learned: Epidemiology of Edward Sanders Low "Tar" Products 5 Exposure Characterization: The "Total Donald Leyden Exposure" Project 6 Biomarkers of the Tobacco-Specific Anthony Tricker N-Nitrosamine 4-(methylnitrosamino)- 1-(3-pyridyl)-1-butanone (NNK) pgNbr=1 2074169493 King, Valerie A. From: Sent: To: Cc: Subject: Mawyer, Denise T. Tuesday, February 29, 2000 11:33 AM Bugg, Joy J.; Patskan, George J.; Kinser, Robin D.; Davies, Bruce D. Cash, Rose B.; Putney, Rebecca M.; King, Valerie A.; McAlpin, Loreen Ground Transportation Arrangements - 3/1-3/3 Listed below are the ground transportation arrangements for the IOM meetings in Washington, DC. Please review and let me know any changes/corrections as soon as possible. Thank you. Wednesday, 3/1 Pick-up Time: Pick-up Location: Participants: Wednesday, 3/1 Pick-up Time: Pick-up Location: Participants: Thursdav. 3/2 Pick-up Time: Pick-up Location: Participants: Friday, 3/3 Pick-up Time: Pick-up Location: Participants: 12:00 noon R&D Exec. Lot Rick Solana George Patskan Don Leyden 6:30 pm Cecil & Ida Green Building (Georgetown) 2001 Wisconsin Ave., NW Washington, DC (202) 334-3588 Rick Solana Don Leyden 3:30 pm R&D Exec. Lot Robin Kinser Bruce Davies 11:30 am Cecil & Ida Green Building (Georgetown) 2001 Wisconsin Ave., NW Washington, DC (202) 334-3588 Robin Kinser Bruce Davies George Patskan 1 pgNbr=2 h-N Ce1~ a-a.y-TD EXPOSURE CHARACTERIZATION The "Total Exposure" Project Donald E. Leyden, Ph.D. V6b69 CVLOZ 2/281oc Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01100 1 pgNbr=3 Machine Cigarette Smoking Compared to Consumer Smoking • Machine smoking » Standardized method useful to assure consistency in manufacturing and inter-brand comparison (e.g. FTC, ISO) » Can provide relative ranking of smoke constituent yields for smoking conditions used • Consumer smoking » Individual smoking styles > e.g. puff volume, frequency, duration » Need biomarker measurements of uptake to determine what smoker exposure is, rather than what it could be 2/28/00 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01/00 2 96b694trL®Z pgNbr=4 What Do We Know from the Literature about Exposure Studies? • Deprivation Studies » that subjects who are deprived of smoking will smoke their next cigarettes differently than those who are not deprived. • Switching Studies » that for a few weeks or months after changing to cigarettes of different yields, smokers have a transient period of adjustment. • Population Studies » that the results of population studies of smokers of cigarettes of various yields smoked ad libitum are inconsistent, but that improved designs for these studies are necessary to address harm reduction. 2128100 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101100 3 96ta691.VLOZ pgNbr=5 What Do We Know about Biologic Sample Matrices to Use for Exposure Studies? • Hair X » Subject to many sources of variation > pigmentation dependent > exposure to ETS > not sufficiently validated > temptingly easy > implies a "time" record • Saliva V- » May be impacted by > time since last smoke > saliva pH > sampling method > easy, OK for smoker classification and has been used a lot (even in quantitative relationships) 2 2sioo Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101100 4 L6V69LtrLOZ pgNbr=6 Model Comparison Model - Saliva cot NICPRED 11.0 10L5 9.5 9.0 fl5ir 8.5 9.0 9.5 iQ0 NI CEd 10.5 11.0 Boswell C, Curvall,M, Elswick Jr.,RK, Leyden,D, Modeling nicotine intake in smokers and snuff users using biological fluid nicotine metabolites, Submitted for Publication. 2/2a/00 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01/00 5 86V69 bbLOZ pgNbr=7 What Do We Know about Biologic Sample Matrices to Use for Exposure Studies? (Cont.) • Serum V » less dependent on external factors than hair/saliva >> cannot provide total smoke constituent uptake » sampling is well controlled but requires clinical environments • Urine V+ » may be-useful to estimate total smoke constituent : uptak »~subject provides sample which makes more ~ natural smoking possible 2128100 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01100 6 661,696ti10Z pgNbr=8 Rosa, et a1.9 1992 • 125 smokers who had smoked preferred brand for >24 weeks (approx. 11 cigt./d) in 4 cigarette 'tar'/nicotine groups • Serum cotinine measured 8 h after last cigarette (HPLC) Rosa, M., Pacifici, R., Altieri, I., Pichini, S., Ottaviani, G., Zucarro, P., How the steady-state cotinine concentration in cigarette smokers is directly related to nicotine intake, Clin. Pharm. Ther., 52: 324-329 (1992) ®®969tiVt®Z 2WOO Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3I01/00 7 pgNbr=9 Rosa et al. (1992) Ntcpti»e intuke and mtFnrna tevrls L®969~trL®Z N~cot~nr fmqldayl Fig. 1. Rclatiunship bcnwccn scrum stcady-slatc cuncentrosion oCcoGnina and daily awailabic nic- otinc (++ = 125. y- = 16.33 + 12.3ar, r= 0.919, P< 0.0001). 2128100 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01/00 8 pgNbr=10 Distribution of Nicotine and Metabolites in Human Urine Compound %(a) % Sum %(b) % Sum 3HC 35 35 36 36 COT-G 17 52 14 50 COT 13 65 9 59 NIC 10 75 9 68 3HC-G 9 84 3 91 NIC-G 3 87 5 96 NNO 7 94 3 99 CNO 4 98 1 100 DMC 2 100 N.D. (Smokers) N = i i 91 a. Byrd GD, Robinson JH, Caldwell WS, deBethizy JD, Nicotine uptake and metabolism in smokers. Paper presented at the 48th Tobacco Chemists' Research Conference, Greensboro, NC, September 25-28, 1994. RSD values calculated to be 21-75% b. Andersson G, Vala ED, Curvall M, The influence of cigarette consumption and smoking machine yields of tar and nicotine on the nicotine uptake and oral mucosal lesions in smokers. J Oral Pathol Med 1997:26:117-123. RSD values calculated to be 28-100% 2128/00 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01/00 9 Z0S696trL®Z pgNbr=11 Urinary Cotinine Model tu1 odell - cot NICPRED 11.0 10.5 lao 9l5 9l0 H 5 -1.- $.5 9.0 9.5 NICEQ 10.0 10.5 11.0 Boswell C, Curvall,M, Elswick Jr.,RK, Leyden,D, Modeling nicotine intake in smokers and snuff users using biological fluid nicotine metabolites, Submitted for Publication. Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 10 £0569 i,VLOZ pgNbr=12 Urinary Nic, Cot, 30H & Conjugates Model - nic, oot, ohc, niog, ootg, ohog Ni4IPRED 11.0 10.5 10.0 9.5 GLo a5i,-. 8.5 9.0 9.5 1Q0 NI CEO 10.5 11.0 Boswell C, Curvall,M, Elswick Jr.,RK, Leyden,D, Modeling nicotine intake in smokers and snuff users using biological fluid nicotine metabolites, Submitted for Publication. zrze o0 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01100 11 b0S69 GVL®Z pgNbr=13 Byrd, et al., 1995 • 33 smokers of their preferred brand (approx. 35 cigt./d) in 4 cigarette 'tar' groups • Urinary nicotine, cotinine, 3-OH-cotinine, and their glucuronide conjugates, and nicotine-N'-oxide*, cotinine-N-oxide*, demethylcotinine * measured in 24-h samples (LC/GC-MS) Byrd, G. D., Robinson, J. H., Caldwell, W. S., deBethizy, J. D. Comparison of measured and FTC-predicted nicotine uptake in smokers, Psychopharmacology 122: 95-103 (1995). * Not determined for the smokers of 1 MG cigarettes. 212a1oo Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01/00 12 50569WOZ pgNbr=14 Data Extracted from Byrd (1995) 100 90 80 70 60 50 40 30 20 10 0 ® Normalized FTC Nicotine  Normalized Daily Nicotine Uptake p Normalized Per Cigarette Nicotine Uptake 1MG ULT Smoker Group FFLT FF v2e/oo Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 13 90569 t-VzaZ pgNbr=15 Byrd, et a1.9 1998 • 72 smokers who had smoked their preferred brand >6 months (approx. 33 cigt./d) in 4 cigarette 'tar'/nicotine groups • Urinary nicotine, cotinine, 3-OH-cotinine, nicotine-N'- oxide, cotinine-N-oxide, demethylcotinine measured in three 24-h samples (LC-MS); salivary cotinine prior to last meal of day (RIA) Byrd, G. D., Davis, R.A., Caldweli, W. S., Robinson, J. H., deBethizy, J. D., A further study of FTC yield and nicotine absorption in smokers, Psychopharmacology 139: 291-299 (1998) 2/28/00 Discusslon Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01100 14 L0969MOZ pgNbr=16 Data Extracted from Byrd (1998) (* Indicates different from FF) 110 100 .-. 0 90 a N s0 ~ C 70  Normalized FTC Nicotine  Normalized Measured Daily Nicotine Uptake 0 Normalized Wasued per Cigarette Nicotine Uptake E e. 10 2MG (<2 mg "tar") i ULT (2.1 - 6.0 mg "tar") FFLT (6.1 -11.9 mg "tar") FF (r_12.0 mg "tar") Smoker Group 2128100 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 15 g05f g LVZQZ pgNbr=17 Dietary Intake is Not an Issue • Philip Morris sponsored a project that determined that dietary nicotine intake is not an issue in using nicotine metabolites as biomarkers for nicotine uptake either by smoking or ETS exposure. Siegmund B, Leitner E, Pfannhauser W, Development of a simple sample preparation technique for the gas chromatographic-mass spectrometric determination of nicotine in edible nightshades (Solanaceae). J Chromatogr A 840(2): 249-260 (1999). Siegmund B, Leitner E, Pfannhauser W, Determination of the nicotine content of various edible nightshades (Solanaceae) and their products and estimation of the associated dietary nicotine intake. J Agric Food Chem 47(8): 3113-3120 (1999). 2128/00 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101l00 16 s®9691~trLoz pgNbr=18 What We Learned • Results from population studies are not consistent between studies, even within the same laboratory • Previous studies can be used to guide the experimental design process for future studies • A majority of population studies have used nicotine and/or its metabolites 2/28/00 Discusslon Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01100 17 0i.S69Lv,LOZ pgNbr=19 vze oa Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01/00 18 44S694VL0Z pgNbr=20 Biomarker Challenges • Uniqueness or nearly so for tobacco smoke • Representative of particulate and gas phase • Representative of health-relevant constituents • Understanding of constituent metabolism • Concentration reflects uptake of cigarette smoke constituent(s) • Based on NRC guidelines of 1986 Benowitz, N.L., Biomarkers of environmental tobacco smoke exposure, Environmental Health Perspectives, 107(2): 349-355 (1999) 212aDO Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101100 19 US69 bbLOZ pgNbr=21 Examples of Potential Smoke Constituents or their Metabolites as Biomarkers of Exposure Particulate Phase • Nicotine Metabolites (More than just cotinine) • 4-aminobiphenyl (adducts) • benzo[a]pyrene (adducts) • PAH (adducts to plasma albumin) • TSNA related biomarkers • solanesol • metals P/28/00 Gas Phase • carbon monoxide (COHb) • cyanide (thiocyanate) • acetonitrile • aldehyde metabolites • 2,5-dimethylfuran Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01l00 20 £b568MLOZ pgNbr=22 Potential Improvements • Potential improvements in experimental design include » » » » smoke constituent, biomarker and biologic medium studied size and diversity of smoker population geographic, ethnic, age, and gender demographics sufficient number of participants to ensure statistical power smoker compliance with test procedures ~ ~ minimize impact on subject smoking behavior determine relationships between cigarette parameters an total exposure results 2128100 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 21 tr49696bLOZ pgNbr=23 .--.. ,-V , Goal of "Total Exposur "Project 2~ U Using biomarker! determine the uptake of vapor phase and particulate smoke constituents for smokers of cigarettes with a range of yields of these constituents to provide a baseline for future cigarette-design related studies. 21281DO Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 22 SLS69PL0Z pgNbr=24 Collaboration Opportunity • What can Philip Morris contribute to this study ~ » » » » Knowledge of cigarette design, construction and testing Expertise in smoke composition and analysis Knowledge of potential biomarkers and their analysis Awareness of the results of previous research in the area • What can other experts contribute to this study? » » Input on identification and aid to prioritize health-relevant smoke constituents and their biomarkers ) especially gas/vapor phase components population study methodologies 2128100 Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 23 K969 6V10Z pgNbr=25 The "Total Exposure Projec~uill contribute to improved understanding of the exposure of smokers to cigarette smoke constituents and establish a baseline for monitoring the impact of new cigarette designs on smokers' exposure. M81oo Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01/00 24 L&Ss694tiLOZ pgNbr=26 02/29/00 TUE 0: 35 FAX 804 274 3730 SCIENTIF AFFAIRS a+s* TX REPORT x~* *a*s~s*a~*ra**:r*a:~ess~s~:* U001 TRANSMISSION OK TX/RX NO 3843 CONNECTION TEL 44780 SUBADDRESS CONNECTION ID PM RICH LEGAL ST. TIME 02/29 10:29 USAGE T 05'57 PGS. 25 RESULT OK FACSIMILE TO: Name. Kevin Osborne Company Philip Morris Fax No. 4-4780 FROM: Valerie Kinlr Philip Morris USA, Worldwide Scientific Affairs, Richmond,'VA Phone: 804/274390~ Fax: 804f274-37;,0 DATE: TUESDAY. p'.8.T3RTJARY 29. 2000 PAGES TO FOLi.OW: 24 This farsimile transmission (and/or the documents accompanying it) may contain cottfidential information belonging to the sender. The information is intended only for the use of the addressee or entity named above. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance of the contents oF this information is strictly prohibited by law. If you have received this transmission in error, please immediately notify us by telephone to arrange for the return of the documents. eQ I MESSAGE: Kevin, Attached is the current draft of Don Levden's nresentation. pgNbr=27 FACSIMILE TO: Name Kevin Osborne Company Philip Morris Fax No. 4-4780 FROM: Valerie King Philip Morris USA, Worldwide Scientific Affairs, Richmond, VA Phone: 804/274-3906 Fax: 804/274-3730 DATE: TUESDAY, FEBRUARY 29, 2000 PAGES TO FOLLOW: 24 This facsimile transmission (and/or the documents accompanying it) may contain confidential information belonging to the sender. The information is intended only for the use of the addressee or entity named above. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance of the contents of this information is strictly prohibited by law. If you have received this transmission in error, please immediately notify us by telephone to arrange for the return of the documents. MESSAGE: Kevin, Attached is the current draft of Don Leyden's presentation. Please let me know any questions or comments you have. Thanks, ® Valerie d , ~ a t9 tn a fD pgNbr=28 -TedS"94,f to. apoS fJffA °~p ! P`'t""t-, - *W"f ~ fa2074169620 ` ~ : pgNbr=29 "Low Yield Cigarettes" March, 2000 aUud ~ - ~~S I P~ -D - /A~ NPU~ Discussion Document Prepared by Philip Morri,~J.JSA for the Institute of Medicine Panel Presentation 3/01 /00 4Z969G'~LCZ `~ 4" pgNbr=30 It should be noted that public health authorities have always maintained that the benefits from quitting smoking greatly outweighed those to be gained through reduction in "tar" zz56s 6VZOz Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=31 Key Assumption Wynder et al, 1957 "It may be predicted that if the average smoker were exposed to only one half the amount of tobacco tar to which the smoker of regular-sized cigarettes is now exposed, his cancer risk would be significantly reduced." Wynder et al., (1957) "A study of tobacco carcinogenesis. II. Dose-response studies," EZ969 11,VL0Z , Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=32 This assertion (driven by the epidemiological data) has been continuously repeated up until the present day in the US and Europe. bZS69LtiLOZ Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=33 "The preponderance of scientific evidence strongly suggests that the lower the "tar" and nicotine content of cigarette smoke, the less harmful would be the effect." "We recommend...the progressive reduction of the "tar" and nicotine content of cigarette smoke." U.S. Public Health Service, "The health consequences of smoking -the changing cigarette," a report of the Surgeon General, 1981. 5Z969 6t7LOZ Discussion Docurnent Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=34 EU - 1999 "However, smokers must be aware that all cigarettes are harmful to health; it is therefore more desirable for them to stop smoking rather than to switch to low- tar cigarettes ...Nevertheless, the existence of a reduced Community ceiling for tar yields ensures a higher level of public health protection than would otherwise be the case..." Proposal for a Directive of the European parliament and of the council, Commission of the European Communities, 1999 9ZS69 4t,LOZ Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=35 During the past 40 years, the U.S. and some international market places have clearly reflected the emphasis on low tar yield cigarettes. zzsss cj7Loz Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=36 What are the Epidemiological Results? gzss9Woz Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=37 Filter versus Non-Filter • 35 studies which allow appropriate relative risks to be determined* • Meta-analytic relative risk for filter versus high tar non-filter cigarettes: - Males RR = 0.58 (95% CI, 0.55-0.62; n = 27) - Females RR = 0.67 (95% CI, 0.59-0.75; n = 14) `~A * Does not include `99 Spanish Study 8ZS6911,b1oZ Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation ~ 3/O1T00 ~~~+ pgNbr=38 High Tar versus Low Tar • The existing data (15 studies) suggest a decrease of 2-3% in the excess risk per mg of tar decrease But.. • No studies are available into the 1990s -----] • No data exist on further tar reductions to 10, 5 or 1 mg/cigarette 0£969 4trLt3Z Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=39 Trends in Lung Cancer Incidence • Some evidence suggests that lung cancer rates have risen faster than might be expected from the smoking trends - Comparison of CPS-1 and CPS-2 b£969LVLOZ Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=40 Workshop models... • Differences in age of smoking initiation • Compensatory smoking • Smoker demography • Possible changes in cigarette composition As discussed in Thun & Heath, (1997) "Changes in Mortality from Smoking in Two American Cancer Society Prospective Studies since 1959." Prev. Med., 26: 422- Z£S69GirCOZ Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=41 However... « ...we cannot tell whether the lung cancer risk might not have increased even more but for the reduced tar yield in cigarettes." Thun & Heath, (1997) "Changes in Mortality from Smoking in Two American Cancer Society Prospective Studies since 1959." Prev. Med., 26: 422- ££969 LVLOZ Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=42 Epidemiology provides the only data that currently permits an assessment of "low yield and disease risk" "Only epidemiologic studies can provide information on modification of the risks of smoking as the cigarette has evolved, and only epidemiologic data can measure the risks of cigarettes under the "natural" circumstances of use. However, the dynamic nature of the exposure challenges the epidemiologic researcher to classify accurately the pattern of cigarette use when changes are made that may not be indexed by tar and nicotine yields measured with a smoking machine." V£569 MOZ Samet, J.M., "Smoking and tobacco control monograph No. 7: Chapter 6- The changing cigarette and disease risk: current status of the evidence." 1996 Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01100 pgNbr=43 Conclusions • Direct epidemiological evidence suggest some reduction in risk for lung cancer • Indirect epidemiological evidence appears to suggest the reverse • Require further research to suggest best way forward • Total exposure measures represent one such important step 9£9694tiLOZ Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 pgNbr=44 Surrogate Markers Suitable for Testing Hans-Juergen Haussmann, Ph.D., ~° INBIFO, Cologne, Germany ~-- 9£9694bLOZ Discussion Document Prepared by Philip Morris, USA, DRAFT for the Inst@ute of Medicine Panel Presentation 25-Feb-0o HHM IOM-Surrogates.ppt 3107100 pgNbr=45 Presentation Objective Discuss surrogate markers that are mechanistically related to and suitable for testing of major diseases with cigarette smoke-related mortality Disease Cigarette Smoke-Related Mortality in US (1990) Cancer 151322 Cardiovascular disease 179820 Respiratory disease 84475 L£5694tiLOZ (Centers for Disease Control and Prevention, 1993) Discussion Document Prepared by Philip Morris, USA, DRAFT for the Instltute of Medicine Panel Presentation 25-Feb-0O HHM IOM-Surrogates.ppt 3/01/00 pgNbr=46 Mechanisms Related to Human Lung Cancer Development • Initiation-promotion paradigm - has not been conclusively applied to lung cancer development - genotoxic events linked to initiation: » activation of proto-oncogenes » inactivation of tumor suppressor genes - reversible morphological changes possibly linked to promotion: » squamous metaplasia irritation - declining lung cancer risk after cessation of smoking indicative of role of promotion 8£969CbLOZ I i 25-Feb-00 HHM IOM - Surrogates.ppt , » basal cell hyperplasia inflammation DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3roUOo pgNbr=47 Experimental Models for Pulmonary Carcinogenesis • Long-term inhalation studies leading to lung tumors - historic attempts: » equivocal, mostly negative, irrespective of rodent species (Coggins, 1997) - recent developments: » Lovelace Respiratory Research Institute studies on rats (Finch et al.) ~ » UC Davis studies on A/J mice (Witschi et al.) » Philip Morris' mechanistic study on rats (Stinn et al.) • Mouse skin painting studies » initiation-promotion model (e.g., Roemer and Hackenberg, 1990) » restricted to particulate phase of cig. Smoke » suitable for specific purposes • Short-term experimental surrogates 25-Feb-00 HHM IOM - Surrogates.ppt DRAFT J ? Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3101/00 6£9694bLOZ pgNbr=48 Requirements for Suitable Surrogate Markers Surrogate markers suitable to assess harm reduction need to be - mechanistically related to the disease, - sensitive and dose-responsive, - reproducible and validated. J oV9ss4Vcoz 25-Feb-00 HHM IOM-Surrogates.ppt DRAFT Discussion Document Prepared by Philip Morris, USA, fortheInst@ute of Medicine Panel Presentation 3/01100 pgNbr=49 Experimental Models for Pulmonary Carcinogenesis • Long-term inhalation studies leading to lung tumors - historic attempts: » equivocal, mostly negative, irrespective of rodent species (Coggins, 1997) - recent developments: » Lovelace Respiratory Research Institute studies on rats (Finch et al.) _ » UC Davis studies on A/J mice (Witschi et al.) » Philip Morris' mechanistic study on rats (Stinn et al.) • Mouse skin painting studies » initiation-promotion model (e.g., Roemer and Hackenberg, 1990) » restricted to particulate phase of cig. Smoke » suitable for specific purposes • Short-term experimental surrogates 25-Feb-00 HHM IOM - Surrogates.ppt DRAFT ? Discussion Document Prepared by Philip Morris, USA, for the tnstitute of Medicine Panel Presentation 3l01100 l.tiS69 LVLOZ pgNbr=50 Long-Term Rat Inhalation Study on Cigarette Smoke and Plutonium Dioxide (incidence of lung tumors; Inhalation Toxicology Research Institute, Annual Report 1995) r-z;:~* male female Hlgh 9moko over-additive effect largely due to impairment of plutonium clearance by cigarette smoke inhalation (Finch et ai., 1998) 25-Feb-00 HHM IOM - Surrogates.ppt DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute oi Medicine Panel Presentation 3/01 /00 ZVSMVLOZ , pgNbr=51 Experimental Models for Pulmonary Carcinogenesis • Long-term inhalation studies leading to lung tumors - historic attempts: » equivocal, mostly negative, irrespective of rodent species (Coggins, 1997) - recent developments: » Lovelace Respiratory Research Institute studies on rats (Finch et al.) _ » UC Davis studies on A/J mice (Witschi et al.) » Philip Morris' mechanistic study on rats (Stinn et al.) • Mouse skin painting studies » initiation-promotion model (e.g., Roemer and Hackenberg, 1990) » restricted to particulate phase of cig. Smoke » suitable for specific purposes • Short-term experimental surrogates 25-Feb-00 HHM IOM - Surrogates.ppt DRAFT r 9 Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3101100 £V9694VLOZ pgNbr=52 Chronic A/J Mouse Inhalation Studies - dose-dependent for ETS surrogate (Witschi, 1998/9) - negative study with mainstream smoke (Finch et al., 1996) - mechanistic issues » postinhalation phase needed for tumor » expression no morphologic changes except increased tumor multiplicity » filtered smoke as effective 0 without postinhalation 50 100 150 TPM Concentration (ug/l) 25-Feb-DO HHM IOM -Surrogates.ppt needs to be further evaluated DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 8/O7100 VV969 4tiLOZ pgNbr=53 Experimental Models for Pulmonary Carcinogenesis • Long-term inhalation studies leading to lung tumors - historic attempts: » equivocal, mostly negative, irrespective of rodent species (Coggins, 1997) - recent developments: » Lovelace Respiratory Research Institute studies on rats (Finch et al.) » UC Davis studies on A/J mice (Witschi et al.) » Philip Morris' mechanistic study on rats (Stinn et al.) • Mouse skin painting studies » initiation-promotion model (e.g., Roemer and Hackenberg, 1990) r » restricted to particulate phase of cig. Smoke » suitable for specific purposes • Short-term experimental surrogates 25-Feb-00 HHM IOM - Surrogates.ppt DRAFT V/ ? Discussion Document Prepared by Philip Morris, USA, for the InstRute of Medicine Panel Presentation 3101100 SV5696tiL0Z pgNbr=54 Philip Morris' Mechanistic Rat Inhalation Study on Combustion Aerosols Inflammatory Cells in Bronchoalveolar Lavage (24 months inhalation, male rats) ~  : ymphocyt es g x g 120 o : o : alwolar nk rte,troph1s i« opr,ages M 40 ~ z *** ~ 100 v r~ 30 80 --1 rn 0 ** W V 20 ~~ J 40~ * cc 10 *** 20--l * ** 1 lirie ~ sham RaSS RA~SS low ~ DNA Adducts, 32P-Postlabeling (18 months inhalation) Male Rats 0 Female rats * I I *** ,t Sham high RASS GROUP high DEE (RASS: Room-aged sidestream smoke; DEE: Diesel engine exhaust, Hock et al., 1999; Friedrichs et al., 2000) 2SFeb-00 HHM IOM - Surrogates.ppt DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3/01/00 9V569 WOZ ~ -- - - -- pgNbr=55 Short-Term Experimental Surrogates ________________________________ _i ;Smoke chemistry ! ;- extended towards IARC carcinogens; -------------------------------I In vitro genotoxicity L - bacterial mutagenicity - mammalian cell mutagenicity V/ V Two-stage transformation (Schiage et al., 1999) I n vitro cytotoxicity r nitiation Promotion In vivo genotoxicity - micronucleus assay (Lee et al., 1990) 25-Fetr00 HHM IOM - Surrogates.ppt LV9694tiL0Z DRAFT Subchronic inhalation L - irritative changes in respiratory tract Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3101 /00 pgNbr=56 Suitability of the Bacterial Mutagenicity Assay to Investigate Cigarette Design Parameters 2000 -I 1000 -I 0 D ~e 0 10 20 30 40 50 60 70 ~ PARAMETER (parameter unitj 25-Feb-00 HHM IOM-Surrogates.ppt DRAFT Test conditions: » cig. smoke condensate » tester strain TA 98, + S9 » multiple linear regression analysis (Tewes et al., 1999) Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3l01/00 9t`969 W®Z --- - - -- i pgNbr=57 Short-Term Experimental Surrogates r_______________________ chemis ry _____ t______________________________ _~ ;Smoke ; ;- extended towards IARC carcinogens; ----- ---------------- In vitro genotoxicity L - bacterial mutagenicity - mammalian cell mutagenicity V/ In vivo genotoxicity - micronucleus assay (Lee et al., 1990) 25-Feb-00 HHM IOM - Surrogates.ppt 6VS69 6ivLOZ Two-stage transformation (Schlage et al., 1999) In vitro cytotoxicity nitiation Promotion J Subchronic inhalation L - irritative changes in respiratory tract DRAFT Disoussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3/01f00 pgNbr=58 Suitability of Subchronic Cigarette Smoke Inhalation to Investigate Morphological Changes in the Rat Respiratory Tract 5 4-1 3 metaplasia, atrophy (e.g., Vanscheeuwijk et al., 1998) consistent throughout ~ respiratory tract (Roemer et al., 1998) 1-1 O 21 days • 90 days Goblet Cell Hyperplasia, Lung r 0 25-Feb-00 HHM 50 100 150 TPM CONCENTRATION (ug/i) I OM - Surrogates.ppt , , , . ~ - dose-dependent K2 Morphological changes: -I 200 DRAFT - hyperplasia squamous reversible (e.g., Coggins et al., 1986) no progression in chronic studies (Haussmann et al., 1998) Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation sioiroo 055694VLOZ - -, pgNbr=59 Cardiovascular Diseases • Types - coronary heart disease/ischemic heart disease » cardiac ischemia » myocardial infarction - cerebrovascular disease » stroke » transient cerebral ischemia - peripheral vascular disease • Mechanisms involved - atherosclerosis - vascular wall changes - vascular tone changes - coagulopathy 25-Feb-00 HHM IOM - Surrogates.ppt DRAF'i' Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3107/00 Eggg9vvLOZ pgNbr=60 The Three Stages of Atherosclerotic Lesion Formation (Breslow, 1996) 25-Feb-00 HHM IOM - Surrogates.ppt Lippproteins DRAFT Hemorrhage & thrnmbosis for the Institute of Medicine Panel Presentation 3/01/00 Z696811,bLOZ - ---- --~ pgNbr=61 Experimental Models for Atherosclerosis (Examples) • ApoE-deficient mouse - atherosclerotic process very similar to human disease in terms of » types and site of lesions (Breslow, 1996) » response to human risk factors, i.e., • high cholesterol / high fat diets • cigarette smoke (subchronic inhalation of sidesteam smoke: Gairola and Daugherty, 1999) » mechanistic hallmarks • lipoprotein oxidation • Rabbits and birds (cockerels, pigeons) - pathogenesis less similar to humans - reported to be responsive to cig. sidestream smoke (Zhu et al., 1993; Penn and Snyder, 1993/1996) 25-Feb-00 HHM IOM - Surrogates.ppt DRAFT ? ? Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3101/00 £5969~VL®Z - pgNbr=62 Chronic Obstructive Lung Diseases • Chronic (obstructive) bronchitis - mediated by pulmonary inflammation (reversible) » increase of bronchoalveolar inflammatory cells • alveolar macrophages • polymorphonuclear leukocytes (PMNL) • lymphocytes - airway obstruction by mucus and morphological changes • Emphysema - irreversible destruction of alveolar walls » disturbance of the protease - anti-protease balance, e.g., • increased elastase activity from inflammatory cells • inactivated protease inhibitors 25-Feb-00 HHM IOM - Surrogates.ppt DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3101/00 V9S69LVL®Z pgNbr=63 Suitability of Subchronic Cigarette Smoke Inhalation to Investigate Chronic Bronchitis I 100 90 80 70 ~ 30-I 20-I 10-I 0_j r 1 10 100 1000 TPM CONCENTRATION (µDq/l) 2~Feb-00 HHM IOM-Surrogates.ppt TtE~FT A : MWS A sub- o : SWS rchronic ~ : RASS J  : RASS chronic .. '.... 4 EN GCE~ r--T_rrrM - dose-dependent increase of absolute and relative number of inflammatory cells (Mull et aL, 1984) - no progression with prolonged inhalation period (Haussmann et al., 1998) Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3101/00 99969 4VL0Z pgNbr=64 Chronic Obstructive Lung Diseases • Chronic (obstructive) bronchitis - mediated by pulmonary inflammation (reversible) » increase of bronchoalveolar inflammatory cells • alveolar macrophages • polymorphonuclear leukocytes (PMNL) • lymphocytes - airway obstruction by mucus and morphological changes • Emphysema - irreversible destruction of alveolar walls » disturbance of the protease - anti-protease balance, e.g., • increased elastase activity from inflammatory cells • inactivated protease inhibitors 25-Feb-00 HHM IOM - Surrogates.ppt DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3/01100 95ggWL®Z pgNbr=65 Experimental Emphysema Models • Chronic inhalation of mainstream smoke (March et al., 1999) - morphological changes similar to those seen in man » » » » increase of alveolar septa mean linear intercept increase of volume density of alveolar air space decrease of volume density of alveolar septa accompanied by accumulation of PMNL • In vitro model for emphysema - inactivation of alpha-l-protease inhibitor (Pryor et al., 1990) 25-Feb-00 HHM IOM - Surrogates.ppt DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3101l00 L99694bL.OZ pgNbr=66 Limited Understanding of Role of Individual Smoke Constituents in Disease Etiology and Mechanisms 1999 SOT symposium: Carcinogenicity of Cigarette Smoke: Bridging the Gap between Complex Mixtures and Individual Components (Burchiel and Witschi) - ... polycyclic aromatic hydrocarbons and the tobacco-specific carcinogen NNK are the most important compounds in cigarette smoke with respect to lung cancer induction in humans (Hecht). ~ - A recently developed tobacco smoke lung cancer model in strain A/J mice ... has made it possible to evaluate the effects of tobacco smoke gas phase in comparison with the effects of full tobacco smoke ... (Witschi) 25-Feb-0O HHM IOM-Surrogates.ppt DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3/01roa 89S69LbLOZ pgNbr=67 Intracellular Interactions of Smoke Constituents Smoke nitric oxide Constituents: NO (gas phase) organic radicals (particulate phase) oxygen~radicals O2 aldehydes (both phases) glutathione "Stress" Pathways: NO-OO- 1 SAPK/JNK 1 OR Toxicity: apoptosis / inflammation genotoxicity irritation / (Mueller et al., 1997; Mueller and Gebel, 1998) cytotoxicity Discussion Document Prepared by Philip Morris, USA, DRAFT for the Institute of Medicine Panel Presentation 25-Feb-00 HHM IOM-Surrogates.ppt 3101/00 69969 6VL®Z pgNbr=68 Summary • Carcinogenicity - short-term markers ~ - skin painting ,/ - long-term inhalation ? • Cardiovascular disease - ApoE-deficient mouse ? - other candidates ? • Chronic obstructive pulmonary diseases - inflammation / - emphysema in vivo ? 69S69~VLUZ 25-Feb-00 HHM IOM-Surrogates.ppt DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute oi Medicine Panel Presentation 3/01/00 pgNbr=69 Conclusion The availability of surrogate assays is restricted by the limited understanding of the relevant pathogenic mechanisms of the diseases. This limitation requires the use of standardized assays as a default. Such are available for carcinogenicity but are lacking for cardiovascular and chronic obstructive pulmonary diseases. Therefore, we have started programs to address these gaps and would welcome guidance from this committee. 25-Feb-00 HHM IOM-Surrogates.ppt DRAFT Discussion Document Prepared by Philip Morris, USA, for the Institute of Medicine Panel Presentation 3/ovoo M694VL®Z I I pgNbr=70 Biomarkers of the tobaccowspecific N-nitrosarnine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) Anthony R. Tricker, Ph.D. Philip Morris Europe Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 Z9B694bL6Z pgNbr=71 In vitro formation of TSNA 4 i N i N' CH3 ~N' H Nicotine Narnicotine N- N=O 'N N=O OH N NNAL !so-NNAL iso-NNAC N" H 'N" H Anabasine Anatabine I I NAB 0H NAT Hecht and Tricker (1999). Nitrosamines derived from nicotine and other tobacco alkaloids. In: JW Gorrod and P Jacob (Eds.) Analytical determination of nicotine and related compounds and their metabolites. Elsevier, Amsterdam. pp. 421-488 Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 89S694tiLOZ pgNbr=72 International Agency for Research on Cancer IARC classifications for TSNA: Group 2B: Possibly carcinogenic to humans 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) N-nitrosonornicotine (NNN) Group 3: Not classifiable as to their carcinogenicity to humans N-nitrosoanabasine (NAB) N-nitrosoanatabine (NAT) International Agency for Research on Cancer (1985). IARC Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. Tobacco Habits Other than Smoking; Betel-Quid and Areca-Nut Chewing; and Some Related Nitrosamines. International Agency for Research on Cancer, Lyon. tr9569 LV L0Z Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3101100 pgNbr=73 Proposed metabolism of NNK NNK-N-oxide (5) 0 N'O . ,I CH3 O N O NNAL (7) Hemoglobin / DNA AA pyridyloxobutylation OH Hemoglobin / DNA methylation 0 r` J~ IIO '--OH N Keto acid (2) OH Hydroxy acid (1) I Diol (4) OH sssssGbLaz Hecht and Tricker, 1999 (simplified) lliscussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/0I/00 oH N'~O 'oH , I N CHz ~ pgNbr=74 In vitro metabolism of NNK in lung and liver Extensive data for rodent tissue metabolism • 0.6-68 % metabolism to NNAL • 15-47 % a-hydroxylation to DNA - reactive intermediates . Pharmacokinetic data for metabolism by a-hydroxylation • a-hydroxylation correlated with DNA adduct formation Limited data for human tissue metabolism • >96.4 % metabolism to NNAL • 0.9 % a-hydroxylation to DNA - reactive intermediates • No pharmacokinetic data • DNA adduct formation requires confirmation No comparative data for rodent / human metabolism Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3101100 999694VLOZ pgNbr=75 Comparative metabolism and adduct formation Comparative in vitro metabolism of [5-3H]NNK, [5 3H]NNAL and [5-3H]NNN and DNA adduct profiles in human, A/J mouse, F344 rat and SG hamster lung and liver under identical experimental conditions • Precision-cut tissue slices in dynamic organ culture • Tissues <1 hr old at start of incubation at pH 7.4 and 37°C for 6 hr • Tissue vitality monitored by LDH leakage and intracellular K+ content • DNA tissue adduct profiles determined by GC/NICI-MS • Michaelis-Menton pharmacokinetics of all major metabolic pathways calculated over a 2 nM - 100 µM substrate concentration range Mainstream smoke uptake predicted to result in 2-10 nM NNK and NNN in lung surfactant layer <114 nM NNAL measured in smoker blood plasma Hecht et aL, Cancer Res., 59:590-596, 1999 Systemic blood TSNA concentrations reach 100 µM in rodents under bioassay protocols Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 L9963HtiLOZ pgNbr=76 Metabolism of 0.1 µM NNK by lung tissues from different species under identical experimental conditions A/J mouse F344 rat SG hamster 1007 Human 75-I Human lung metabolism of NNK • Significantly different to rodents • Limited metabolic activation by a- hydroxylation to keto acid or HPB • Limited detoxification by pyridine N-oxidation • NNK reduced to NNAL (>97%) a U 25-I NN< 3HwaterKeto acid 5 10 15 20 25 30 35 40 Retertirn Time (min) Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 s9s69Gbzoz pgNbr=77 Potential biomarkers of NNK exposure NNK-N-oxide (5) Not excreted in uma s ~O H CH3 0, Minor metabolite OH N'O i N CH3 N 0 NNAL (7) Hemoglobin / DNA pyridyloxob utylati on Metabolites not specific to NNK i ~(' O N.CH OH OH . NNAL-N-oxide OH C ~ Hemoglobin I DN~ I ' methylation O ~ OH I IIO N o ac OH [ OH y rid ( io NNAL-Gluc Hecht and Tricker, 1999 (simplified) Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/01/00 69969417LOZ 0 HOO HO~ O HO- OH pgNbr=78 Proposed metabolism of NNN myosa,lno 0 ~ ^ x A 'N=NOHJ s C~vv o C ! ~ }l ^ 'OH CO~ v XO N koto acid 0 ~ Hemoglobin I DNA pyridyloxobutylatio n OH 'OH ;N vai va laelotw O O N hydroxy acid Hecht and Tricker, 1999 Discussion Document Prepared by Philip Morris, USA for the Institute of Medicine Panel Presentation 3/Ol/00 JH O O OH N laetol OH OL5691,VL®Z ; pgNbr=79 Biomarkers of NNK exposure: urinary metabolites Smoking status ETS exposure (hours/week) Cotinine (mg/day) NNAL + NNAL-Gluc (pmol/day) NNK equiv. (µg/day) NS 3.6±6.5 0.018±0.013 ~ 15000 10000 5000 0 TA98 NA - Not active Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation, 3/01/00 TA100 NA NA TA1535 TA1 537 09 1 R4F  EHC 6Q969 W(BZ pgNbr=118 DRAFT Ames Assay -S9 1600 , 1400 1200 - •~ 1000 ~ 71- 1 R4F  EHC 400 ~ 200 ~.~ NA ~ NA NA NA NA TA98 TA100 TA1535 TA1537 NA - Not active Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation, 3/01/00 o G969 ~VLOz pgNbr=119 DRAFT lPM Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation, 3/01/00 Cytotoxicity GVP 0 1 R4F  EHC 9.a969dtrLOZ pgNbr=120 DRAFT Inhalation • The effects observed in animals exposed to smoke from the EHC were similar to those typically observed in animals exposed to the smoke from -Eeavenzional cigarettes, ex. 1 R4F. / r;n~ c,4~ -6 ,. uj4442&,L i Discussion Document Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation, 3/01/00 Z b969 WZ®Z pgNbr=121 body weight respiratory rate lymphocyte count liver enzymes rel. organ weights changes in nose changes in tarynx i changes in trachea and lung O Rcl. Biological Activity (% of 1R4F) N C ~ C O pgNbr=122 ~ How Does the Uptake of Gas Phase and Particulate Phase Smoke Constituents Relate to Cigarette Design ? The "Total Exposure" Project .0 Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3107/00 1 t7~969 6'Gl-oZ pgNbr=123 Machine Cigarette Smoking Compared to Consumer Smoking • Machine smoking » Standardized method useful to assure consistency in manufacturing and inter-company comparison (e.g. FTC, ISO) » Can provide relative ranking of smoke constituent yields • Consumer smoking » Individual smoking styles > e.g. puff volume, frequency, duration » Need biomarker measurements of uptake Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01/00 2 9 6969 bVL0Z pgNbr=124 Rosa, et al., 1992 • 125 smokers who had smoked preferred brand for >24 weeks (approx. 11 cig./d) in 4 cigarette 'tar'/nicotine groups • Serum cotinine measured 8 h after last cigarette (HPLC) Rosa, M., Pacifici, R., Altieri, L, Pichini, S., Ottaviani, G., Zucarro, P., How the steady-state cotinine concentration in cigarette smokers is directly related to nicotine intake, Clin. Pharm. Ther., 52: 324-329 (1992) Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3I01I00 3 91.9s9H71oZ pgNbr=125 Results from Rosa, et al., 1992 V()1.W44.5J nuMXC.x9 Naneinc lxemkc nnd evtiniasr kr+clt 327 NfcaNne (mg/day} l. Fidstlenahdp betwtcn vrsvin ntndys4~te cuwmntrxfiun W cudins mE dqAY eraA®hle nic- oUnc (n - 125, Y a 16.33 t 17_Nlr. r= 0.919. p< D.000J). Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 4 LL9696VLOZ pgNbr=126 Pp. . r~V'~' RNi....t „0, lwimr, ,~rAh ~i 1. , Byrd, et aI ®, 1995 I • 33 smokers of their preferred brand (approx. 35 cig./d) in 4 cigarette 'tar' groups • Urinary nicotine, cotinine, 3-OH-cotinine, and their glucuronide conjugates, and nicotine-N'- oxide*, cotinine-N-oxide*, demethylcotinine * measured in 24-h samples (LC/GC-MS) Byrd, G. D., Robinson, J. H., Caldwell, W. S., deBethizy, J. D. Comparison of measured and FTC-predicted nicotine uptake in smokers, Psychopharmacology 122: 95-103 (1995). * Not determined for the smokers of 1 MG cigarettes. Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 5 81•9696vLOZ pgNbr=127 Data Extracted from Byrd (1995) Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 6 6456JW®Z pgNbr=128 Byrd, et aIe9 1998 • 72 smokers who had smoked their preferred brand >6 months (approx. 33 cig./d) in 4 cigarette `tar'/nicotine groups • Urinary nicotine, cotinine, 3-OH-cotinine, nicotine-N'-oxide, cotinine-N-oxide, demethylcotinine measured in three 24-h samples (LC-MS); salivary cotinine prior to last meal of day (RIA) Byrd, G. D., Davis, R.A., Caldwell, W. S., Robinson, J. H., deBethizy, J. D., A further study of FTC yield and nicotine absorption in smokers, Psychopharmacology 139: 291-299 (1998) Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101100 7 OZ969 4VLOZ pgNbr=129 Data Extracted from Byrd (1998) FTC Nicotine, Measured Daily Nicotine Uptake, and per Cigarette Nicotine Uptake (* indicates different from FF) 2MG (<2 mg "tar') ULT(2.1-6.0mg'Yat') FFLT(6.1-11.9mg"taP') FF(>=12.0mg"tar') Smoker Group Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 8 bZ969 WOZ pgNbr=130 Summary of Results • Results from population studies are not consistent between studies, even within the same laboratory • Results from "switching" studies provide information on transient changes • Various biomarkers have been used • A majority of population studies have used nicotine and/or its metabolites • These studies can be used as guides or pilots for the design of future studies Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 9 zz9s9~tiL0z pgNbr=131 Biomarker Challenges • Challenges for biomarkers » uniqueness for tobacco smoke » representation of particulate and gas phase » representation of health-relevant constituents » understanding of constituent metabolism » concentration reflects uptake of cigarette smoke constituent(s) » based on NRC guidelines of 1986 Benowitz, N.L., Biomarkers of environmental tobacco smoke exposure, Environmental Health Perspectives, 107(2): 349-355 (1999) Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/D1/00 10 EZ9696VLOZ pgNbr=132 Potential Im provementa • Potential improvements in experimental design include » smoke constituent, biomarker and biologic medium studied » size and diversity of smoker population > Geographic, Ethnic, Age, Gender demographics > sufficient number of participants to ensure statistical power » smoker compliance with test procedures » collect information that could be useful to address relevant genetic polymorphism Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01100 11 VZ969 6VLQZ pgNbr=133 Goal of Proposed "Total Exposure" Project Using biomarkers determine the uptake of vapor phase and particulate smoke constituents for smokers of cigarettes with a range of yields of these constituents to provide a baseline for future cigarette- design related studies. Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101100 12 qz96stiVLOz pgNbr=134 Definition of "Total Exposure" • Addresses both number and type of cigarettes smoked • Include both particulate phase (PP) and gas/vapor phase (VP) constituents • Exposure defined as average daily uptake in a `steady state' situation • Includes exposure to mainstream smoke, ETS, and other sources Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101/00 13 szsssWoz pgNbr=135 Steps • Determine smoke constituents and biomarkers to use » Opportunity for input from health science community • Identify validated analytical methods for measuring smoker exposure » Opportunity for suggestions or information on newly developed methods • Predetermine the criteria for measures of significance • Determine relationship between cigarette parameters and total exposure Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101100 14 LZ9C96bLOZ pgNbr=136 Required Ancillary Information • Number of cigarettes smoked/day • FTC tar and nicotine rating of cigarettes smoked • Diary/questionnaire information on how test population smokes • Use objective measures wherever possible to supplement subject information Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101100 15 8Z969bbL®Z pgNbr=137 Sampling and Analysis Methodologies • No impact on subject smoking patterns • High level of subject compliance • Accepted and validated analytical methodology • Establishment of analytical and biological variation (inter-personal, intra-personal) • Data analysis with predetermined methods and research hypotheses Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3101100 16 szsssat,toz pgNbr=138 Collaboration Opportunity • What can Philip Morris contribute to this study » Knowledge of cigarette construction and testing >> Expertise in smoke composition and analysis • What can other experts contribute to this study? » Input on identification and aid to prioritize health- relevant smoke constituents and their biomarkers > especially gas/vapor phase components » population study methodologies Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01/00 17 0£969W7L0Z pgNbr=139 d.d d.d L b aME"! !!Y The "Total Exposure Project" will contribute to improved understanding of the exposure of smokers to cigarette smoke constituents and establish a baseline for monitoring the impact of new cigarette designs on smokers' exposure. Discussion Presentation Prepared by Philip Morris USA for the Institute of Medicine Panel Presentation 3/01l00 18 ~MssGVL®z pgNbr=140