Disclaimer: The information in these materials is a formal dissemination of information by BAT and issues discussed in my presentation are not in any official manner related to/voiced by EUROTOX or its member societies. Electronic cigarettes: Pre-clinical and clinical assessment Dr Chris Proctor Chief Scientific Officer, British American Tobacco Tuesday 12 th September 2017
Conflict of interest Dr. Proctor is employed by British American Tobacco Company (BAT). The research reported was funded by BAT. I declare that this work was fully funded by British American Tobacco (Investments) Ltd and that myself and my coworkers were full time employees of British American Tobacco (Investments) Ltd for the duration of the research.
AGENDA 1. Background 2. Assessing the risk profile of e-cigarettes 3. Product bridging 4. Summary
E-cigarettes have evolved rapidly Development of e-liquids
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Our global credibility for assessing e-cigarettes (1): Presentations at regulatory meetings US, FDA Proctor CJ (2015). Lessons from clinical studies using biomarkers of exposure to assess toxicant exposure. Presentation at Biomarkers of Tobacco Exposure: a public workshop, Washington DC, 3 rd 4 th Aug 2015 Proctor CJ (2016). Utility of Biomarkers of biological effect as end-points for the disease relevant assessment of novel tobacco and nicotine products as potentially reduced risk products presentation at Biomarkers of biological effect: A public workshop Session 5 FDA/CTP Scientific Workshop Washington 5 th April 2016 US, National Academy of Sciences Murphy J (2017). National Academy of Sciences, Engineering and Medicine; Committee on the review of the health effects of Electronic Nicotine Delivery Systems: Clinical studies in ENDS users. Washington 21 st February 2017 Gaça M (2017). National Academy of Sciences, Engineering and Medicine; Committee on the review of the health effects of Electronic Nicotine Delivery Systems: Results from in vitro assays. Washington 21 st February 2017
Our global credibility for assessing e-cigarettes (2): Publications Accepted Murphy, JJ (2017). PROFILE: British American Tobacco discusses the impact of smoking and the potentially reduced risks of e-cigarettes. Pan European Networks, Government vol 21, 52-55. Costigan S, et al. (2015). An approach to ingredient screening and toxicological risk assessment of flavours in e-liquids. Reg Tox Pharm, 72: 361-369, DOI:10.1016/2015.05.018 Rawlinson C, et al. Chemical Characterisation of Aerosols Emitted by Electronic Cigarettes Using Thermal Desorption Gas Chromatography Time of Flight Mass Spectrometry. Journal of Chromatography A. 1497: 144-154 DOI: 10.1016/j.chroma.2017.02.050 Margham J et al. (2016). Chemical composition of aerosol from an e-cigarette: a quantitative comparison with cigarette smoke. Chemical Research in Toxicology 29: 1662-1678; DOI: 10.1021/acs.chemrestox.6b00188 Adamson J et al. (2016). Application of dosimetry tools for the assessment of e-cigarette aerosol and cigarette smoke generated on two different in vitro exposure systems. Chemistry Central Journal 10: 74. DOI 10.1186/s13065-016-0221-9 Thorne D et al. (2016). The mutagenic assessment of an electronic-cigarette and reference cigarette smoke using the Ames assay in strains TA98 and TA100. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 812: 29-38. http://dx.doi.org/10.1016/j.mrgentox.2016.10.005 Comprehensive published Azzopardi D et al. (2016). Electronic cigarette aerosol induces significantly less cytotoxicity than tobacco smoke, Toxicology Mechanisms and Methods 26: 477-491; DOI: 10.1080/15376516.2016.1217112 Cunningham A et al. (2016). Development, validation and application of a device to measure e-cigarette users puffing topography. Scientific Reports 6, 35071; doi: 10.1038/srep35071. Fearon IM et al. (2017). E-cigarette Nicotine Delivery: Data and Learnings from Pharmacokinetic Studies. American Journal of Health Behaviour 41: 16-32. DOI: https://doi.org/10.5993/ajhb.41.1.2 data set gathered on our Taylor M et al. (2016). E-cigarette aerosols induce lower oxidative stress in vitro when compared to tobacco smoke. Toxicology Mechanisms and Methods 26: 465-476; DOI: 10.1080/15376516.2016.1222473 Thorne D et al. (2017). The comparative in vitro assessment of e-cigarette and cigarette smoke aerosols using the γh2ax assay and applied dose measurements. Toxicology Letters 265: 170 178. http://dx.doi.org/10.1016/j.toxlet.2016.12.006 closed modular e-cigarette Banerjee, A., Haswell, L.E., Baxter, A., Parmar, A., Azzopardi, D., Corke, S., Thorne, D., Adamson, J., Mushonganono, J., Gaca, M.D. and Minet, E., 2017. Differential Gene Expression Using RNA Sequencing Profiling in a Reconstituted Airway Epithelium Exposed to Conventional Cigarette Smoke or Electronic Cigarette Aerosols. Applied In Vitro Toxicology, 3(1), pp.84-98. Hill, A. and Camacho, O.M., 2017. A system dynamics modelling approach to assess the impact of launching a new nicotine product on population health outcomes. Regulatory Toxicology and Pharmacology, 86, pp.265-278. Haswell, L.E., Baxter, A., Banerjee, A., Verrastro, I., Mushonganono, J., Adamson, J., Thorne, D., Gaça, M. and Minet, E., 2017. Reduced biological effect of e-cigarette aerosol compared to cigarette smoke evaluated in vitro using normalized nicotine dose and RNA-seq-based toxicogenomics. Scientific Reports, 7(1), p.888. Breheny, D., Oke, O., Pant, K. and Gaça, M., 2017. Comparative tumor promotion assessment of e cigarette and cigarettes using the in vitro Bhas 42 cell transformation assay. Environmental and Molecular Mutagenesis, 58(4), pp.190-198. Taylor, M., Jaunky, T., Hewitt, K., Breheny, D., Lowe, F., Fearon, I.M. and Gaca, M., 2017. A comparative assessment of e-cigarette aerosols and cigarette smoke on in vitro endothelial cell migration. Toxicology Letters, 277, pp.123-128. ------------------------------------------------------------------- Submitted (under review) Murphy J et al. Assessing modified risk tobacco and nicotine products: Description of the scientific framework and assessment of a closed modular electronic cigarette. Reg Tox Pharma (Submitted)
In the UK there is a growing consensus on e-cigarette harm reduction potential Kevin Fenton, Public Health Director of Health and Wellbeing: The wider body of evidence consistently finds that e-cigarettes are less harmful than smoking E-cigarettes: an evidence update The current best estimate is that e-cigarettes are around 95% less harmful than smoking Nicotine without smoke: tobacco harm reduction Promote e-cigarettes widely as substitute for smoking says new RCP report Electronic cigarettes (also known as vapourisers) Compared to tobacco products, electronic cigarettes are significantly safer
Consumer safety testing of e-cigarettes What do we need to know? eliquid What s in the liquid? Device What s the device made of? Does it conform to electrical safety? Product What s in the vapour? How stable is the product over time? How is it used? 1. Costigan and Meredith (2015) Regul Toxicol Pharmacol 72:361 369
A three-step scientific journey to substantiate reduced risk 01 E M I S S I O N S 02 E X P O S U R E 03 R I S K Chemical studies in laboratories using smoking/vaping machines comparing NGPs with conventional cigarettes. Provides evidence of reduced hazard and RRP potential. Cannot represent range of human exposures. Studies demonstrating switching to NGPs results in lower exposure to toxicants by making comparisons with continued smoking and quitting. Studies demonstrating changes on switching to NGPs on biomarkers of biological effect, supported by toxicology, systems science, modelling and post market studies. Provides evidence to reduce risk at both the individual and population level.
A three-step scientific journey to assess the risk profile of e-cigarettes 01 E M I S S I O N S Untargeted emissions Targeted emissions Environmental emissions 02 E X P O S U R E Puffing behaviour Average daily consumption Clinical PK Clinical BoE REDUCED TOXICITY IN LAB MODELS 03 R I S K Clinical BoBE Risk perception Post market surveillance in vitro reg tox in vitro disease models in vitro systems science 2. Murphy, JJ (2017). Pan European Networks, Government 21:52-55 and 3. Murphy J et al (2017) Reg Tox Pharm (Submitted)
Origin of Harmful and Potentially Harmful Constituents (HPHC) PRODUCT TOBACCO PRESENT AEROSOL FORMATION MECHANISM CIGARETTE Yes Combustion & pyrolysis of tobacco NUMBER OF COMPOUNDS IN AEROSOL TYPICAL NUMBER OF TOXICANT TYPES HPHC FORMATION MECHANISMS >7000 100 150 Transfer from tobacco Pyrosynthesis of tobacco UNTARGETED EMISSIONS 2 ENDS No Vaporisation of e-liquid 10 100 <5 Poorly stewarded e-liquids (eg. containing contaminants, CMRs) Thermal degradation of humectants ( Dry wicking ) Extractables & Leachables from device during storage or heating 4. Rawlinson et al. (2017) J Chrom A, 1497: 144 154
Topography studies and machine puffing regimes 5. Cunningham et al. (2016) Sci Rep 6: 35071X 6. Prasad et al., in preparation; 7.CORESTA recommended method number 81 (2015) https://www.coresta.org/sites/defa ult/files/technical_documents/main /CRM_81.pdf (accessed 17th Feb 2017) Key findings: Studies show that consumer behaviour with ENDs is in-line with CORESTA recommended method N o 81 (CRM81) 5,6 : 55mL puff volume/3 s puff duration/30s interval between puffs 7
E-cigarette has reduced levels of toxicants relative to cigarettes* 8. Margham et al (2016) Chem Res Tox, 29: 1662 1678 Substantial reductions in toxicant levels observed versus cigarettes for all regulatory lists under these test conditions *These qualities do not necessarily mean this product produces less adverse health effects than tobacco products
E-cigarette has reduced toxicity relative to cigarettes (1)* OECD* TG 471: Bacterial Reverse Mutation Test, S. typhimurium TA98 E-cigarettes gave no response even at 900 puffs Exposure to reference cigarette smoke caused mutations in a dose dependent manner; e-cigarettes gave no response 9. Thorne D et al (2016) Mutation Research/Genetic Toxicology and Environmental Mutagenesis 812: 29-38 E-cigarettes gave no response even after 900 puffs 10. Thorne D et al -submitted. *These qualities do not necessarily mean this product produces less adverse health effects than tobacco products
E-cigarettes have reduced responses in disease relevant models 11. Taylor M et al. (2016). Toxicology Mechanisms and Methods, 26: 465-476 12. Taylor M et al. (2017). Toxicology Letters, 277, 123-128. 13. Breheny, et al. (2017). Environmental and Molecular Mutagenesis, 58(4), 190-198. 14. Thorne D et al. (2017). Toxicology Letters, 265: 170 178. 15. Azzopardi et al (2016) Toxicol. Mech. Methods 26:477 491 Substantial reductions in responses in tests relevant to oxidative stress, CVD, genotoxicity, tumour promotion and cytoxicity vs. cigarettes *These qualities do not necessarily mean this product produces less adverse health effects than tobacco products
Irritancy assessment of aerosols* Comparison of cytotoxicity after cigarette and e-cigarette exposure using EpiAirwayTM IRRITANCY (WHOLE AEROSOL) No cytotoxicity with e-cigarette exposed EpiAirway TM Schematic representation of the VITROCELL VC 01 Smoking Robot, mammalian 12/6 CF stainless-steel exposure module, and EpiAirway TM tissue model. (A) VC 01 single port smoking robot, enclosed in a ventilation hood with a piston/syringe that draws and delivers smoke or aerosol to the dilution bar. (B) Dilution bar, where smoke or aerosol is diluted, mixed, and delivered to the exposure module. Diluted smoke/aerosol within the dilution bar transits to exhaust. (C) 12/6 CF stainlesssteel exposure module, where EpiAirway TM inserts are housed during exposure. (D.I) Culture insert on which EpiAirway TM tissue culture is supported at the air liquid interface with smoke/aerosol distributing trumpet sitting 2 mm above the surface of the tissue. (D.II) EpiAirwayTM human airway epithelium. (D.III) Fresh culture media (AIR-100 maintenance media) basally feeding human airway epithelium. Transmission electron micrograph (magnification 20,000) showing (E.I) cilia and (E.II) tight junctions. Haematoxylin and eosin stained cross-sections (magnification 360) of (E.III) pseudostratified mucocilary morphology of EpiAirway TM tissue and (F) excised human bronchial epithelium for comparison. 16. Neilson et al (2016)Toxicol. In Vitro; 29(7):1952 1962 *These qualities do not necessarily mean this product produces less adverse health effects than tobacco products
Comparing transcriptional perturbations in MucilAir * 48, 854 genes/ RNA features screened 3R4F 8197 significant genes/rna features Vype epen 49 significant genes/rna features Vype epen* 113 significant genes/rna features RNA-seq data mapped onto 131 pathway-focussed gene sets with specific biological function and disease processes 3R4F cigarette smoke 1/30 epen aerosol 1/7 epen aerosol 1/3 3R4F e-cig e-cig* 3R4F e-cig* Toxicogenomics RNA-seq differential gene expression e-cig * X2 nicotine dose 17. Haswell, L.E. et al (2017) Sci Reports, 7(1), 888. Gene enrichment analysis: heatmap indicating fold change for RNAs significant at pfdr<0.05 *These qualities do not necessarily mean this product produces less adverse health effects than tobacco products
Adverse Outcome Pathways (AOPs) Describes a sequential chain of causally linked events at different levels of biological organisation, that lead to an adverse effect 18. Ankley et al (2010) Environ.Toxicol.Chem 29(3): 730 741 Collaboration to Support AOP Build Two AOPs (BAT/PMI/SELVENTA) Oxidative Stress Leading to Hypertension https://aopwiki.org/aops/149 19. Lowe et al. (2017) Applied In Vitro Toxicology 3(1): 131 148 EGFR Activation Leading to Decreased Lung Function https://aopwiki.org/aops/148 20. Luettich et al.(2017) Applied In Vitro Toxicology. 3(1): 99 109
Nicotine exposure: PK studies epen [1] E-cigarette nicotine delivery: data and learnings from pharmacokinetic studies :21; [2] Product used in both studies: epen closed modular ENDS, [nic] =18mg/ml Key findings 1st study: Smokers (naïve ENDS users) have reduced nicotine uptake with ENDS in comparison to cigarettes 2nd study: Smokers (experienced vapers) may adapt their puffing behaviour and therefore have similar nicotine uptake with ENDS in comparison to cigarettes 21. Fearon et al (2017) Am J Health Behav 41(1): 16-32
Clinical study design to assess exposure from e-cigarettes use in comparison to smoking and cessation 5 day, randomised forced switch, confinement study Typically 30 subjects/arm Measure Biomarkers of Exposure (BoE) levels in ENDS users relative to smoking and cessation Cessation arm objectives (i) maximum effect comparator and (ii) baseline for confounders Subject Original product Randomisation to Switch product Baseline Day 1 Day 2 Day 3 Day 4 Day 5 Smoker Cigarette Cigarette Smoker Cigarette ENDS Smoker Cigarette Cessation Forced switch = Sampling point 22. Gale et al (2017) BMC Public Health,
Public Health clinical studies on e-cigarettes Roswell Park Cancer Institute 23 Biomarker Average % reductions EoS v baseline COHb (CO) 63 75% NNAL (NNK) 57 64% 3-HPMA (Acrolein) 49 56% 23. Goniewicz et al (2017) Nic and Tob Res 19(2), 160 167 Conclusions Solus ENDS use can reduce exposure to selected carcinogens and toxicants to similar levels as cessation Exposure reductions with dual use are lower than solus use Results corroborated with recent 6 month BoE study sponsored by Cancer Research UK : Shahab et al (2017) Annals of Int Medicine 166:6, pp. 390 400
Population risk: studies in post market surveillance ER ESTIMATES X POPULATION USAGE = POPULATION RISK IMPACT Excess Risk (ER) estimates determined from pre-clinical and clinical assessment Risk estimates calculated for: Smoking Solus ENDS use Dual use with respect to never smokers and Nicotine Replacement Therapy Usage of products assessed across population: Smoking Solus ENDS use Dual use Non smokers Quitting Dynamic model 24,25 to assess population risk impact 24. Hill & Camacho (2017) Reg Tox Pharma 86, pp.265-278. 25. Model design to meet FDA expectations (FDA 2012) http://www.fda.gov/downloads/tobaccoproducts/guidancecomplianceregulatoryinformation/ucm297751.pdf. (Accessed 20 th February 2017).
Product bridging Conundrum: can we bridge data between product variants in the fast paced world of next generation products? Yes this is happening today in the world of similar and bio-similars Modified pharma industry approach to bridging could be applied in the nicotine and tobacco product context Foundation datasets on the original product variant ( reference ) can be added to on a need basis to allow bridging to the new variant ( similar )
Proposed principles of product bridging
SUMMARY Scientific framework established to substantiate risk reduction potential of e-cigarettes Pre-clinical assessment shows that e-cigarettes have the potential to reduce risk relative to cigarettes All data is being published in peer review literature A workable framework for bridging data between product variants is required
Disclaimer: The information in these materials is a formal dissemination of information by BAT and issues discussed in my presentation are not in any official manner related to/voiced by EUROTOX or its member societies.