Testing For Safety & Efficacy Alain Khaiat PhD Vice President R&D Johnson and Johnson Asia Pacific

1. Testing For Safety & Efficacy Alain Khaiat PhD Vice President R&D Johnson and Johnson Asia Pacific

2. Testing for Safety • Cumulative Irritation: • To determine the irritation potential of topical product left on skin. • Sensitization (RIPT): • To determine the potential of a product to cause allergic contact response. • Photo-toxicity: • To assess the potential of ingredients to causelocalized irritant reactions when exposed to sunlight • Photosensitization: • To determine the potential of a product to cause allergic contact response when exposed to sunlight.

3. Safety Testing Cumulative Irritation • 25 volunteers • Occlusive patch applied for 48 to 72 hours RIPT - Predictive testing for skin sensitizers. • 100 - 200 volunteers • 10, 48 hr Patches - 3 weeks • Rest period - 2 weeks • Challenge 48hr at new skin site.

4. Safety Testing Phototoxicity • 20 volunteers • 1 dose • 0, 24, hr exposure & patches • Read @ 48, 72 hr Photoallergy • 25 volunteers, fair skinned • Induction phase – twice/week for 3 weeks. Reading & irradiated 24 hrs after patching • Rest period – 2 weeks • Challenge phase – Patch at new skin site. Read @ 24, 48, 72 hrs

5. EYE MILDNESS

6. Suggested Test Criteria EC50TEP Rating 2.20 or higher PASS 1.8 - 2.19 Borderline 1.79 or lower FAIL Ocular Safety In-vitro Screening: Transepithelial Permeability (TEP) EC50= test pdt concn where 50% of the max amount of dye to leak through the barrier into the lower well. Corneal Epithelium MDCKCells • MDCK cells grown to confluence • Tight junctions b/w cells form an impermeable barrier • Dye marker will not cross undamaged cell layer

7. Required Tests Criteria Ocular Safety In-Vivo Test: Human Ocular Irritation Test Stinging at time of installation Which is not statistically higher than sterile water control • Panel of 10 subjects • Supervised by an opthalmologist • Subject focus on any minor discomfort • in their eyes after application of warmed • test products. • Measures sting, lacrimation, irritation • One eye receives 10% conc of test substance and the other eye receives distilled water When 5 subjects (or more) experience larger responses on test product than to water, the test product will generally fail to be “non – irritating” to the eye

8. MOISTURIZATION

9. Moisturization Test Subjects: • Female adults, usually 20 to 55 yrs old • In good health, no allergy to cosmetics • Normal to Dry skin

10. Moisturization Test Design: • Base size of 10 to 20 • Randomized • Control – untreated skin and/or benchmark

11. Moisturization Test Site: • 4cm x 8cm rectangle on volar forearm • Usually 2 test sites per arm Product Usage: • 0.1mL per test site

12. Moisturization Before the test: • No leave-on products 24 hours before • No caffeinated drinks 1 hour before • 30-minute acclimatization to room conditions

13. Moisturization Instrumental Evaluations: • Baseline, 1, 4, 8, (12), (24) hours • Skicon® – Conductance • Tewameter® – TEWL • Visioscan® – Surface roughness

14. Moisturization Success Criteria: • Increase in conductance vs. baseline • Higher increase vs. control/benchmark • Reduction in TEWL vs. baseline • Higher reduction vs. control/benchmark

15. Moisturization Other Considerations: • For wash-off products, need to do longer term studies e.g. 2 weeks

16. Suggested Tests/Data Instrumental Skicon (based on conductance) Corneometer (based on capacitance) TEWAmeter Criteria Increase in parameter versus baseline, untreated skin or benchmark (0, 2, 4, 6, 8, 24 hours, weeks) Criteria Decrease in trans-epidermal water loss versus baseline, untreated skin or benchmark (0, 2, 4, 6, 8, 24 hours) Claim: Moisturizes, hydrates

17. Suggested Tests/Data Derm Assessment Dryness Scaling Erythema Instrumental Skicon/Corneometer TEWAmeter Erythema Index Digital Photography Self-Assessment Irritation Criteria Improvement in grading vs. baseline, untreated skin or benchmark Improvement in parameter vs. baseline, untreated skin or benchmark Improvement in grading vs. baseline, untreated skin or benchmark Claim: Prevents symptoms of dry skin

18. Claims related to products which treat or prevent Acne and Pimples

19. Non-comedogenic Oil Free Contains a blackhead fighting medicine Long lasting oil control Prevents future blackheads Won’t overdry your skin Reaches deep down into pores Removes oil, dirt and make-up

20. Acne Test Subjects: • Female adults, usually 20-35 yrs old • In good health, no allergy to cosmetics • Mild to Moderate Acne

21. Acne Test Design: • Base size of 30 to 50 • Double-Blind, Randomized • Control – vehicle and/or benchmark

22. Acne Test Site: • Whole face Product Usage: • Normal or habitual use • Usually 2x daily

23. Acne During the study: • Use of skin care products with active ingredients NOT allowed

24. Acne Clinical Evaluations: • Baseline, 1, 2, 4, 8 weeks • Overall Acne Severity (Cook’s Grading Scale) • Lesion Count – papules, pustules, open & closed comedones

25. Acne Instrumental Evaluations: • Skinskan® – inflammation • DRS – erythema • PRIMOS® – size & elevation • Photography – overall improvement

26. Acne Success Criteria: • Improvement vs. baseline • Better improvement vs. vehicle • Better improvement vs. benchmark Other Considerations: • Inclusion of safety assessment e.g. erythema, edema, dryness, scaling

27. Suggested Tests/Data The product contains a keratolytic agent Self assessment (150-based consumer research) Instrumental Measures and Derm assessment. Criteria E.g. Salicylic Acid 0.5-2.0% Statistically significant results vs baseline/control/ benchmark Digital photography. Numerical count of lesions, Dermatological Grading (versus visual standards). Claim: Helps treat and prevent pimples and blackheads

28. Example of Dermatologist Grading Scale:Modified Cook’s Grading scale

29. Suggested Tests/Data The product contains a keratolytic agent, exfoliant or absorbent. Self assessment (150-based consumer research) Criteria E.g. Salicylic Acid, AHA, Benzoyl Peroxide, Polyethylene, Silica, Kaolin. Statistically significant results vs baseline/control/ benchmark Claim: Keeps pores unclogged

30. Suggested Tests/Data The product does not contain any ingredient that would be considered an oil. Criteria The CTFA/INCI ingredient listing for each ingredient does not contain the word “oil”. Claim: Oil-Free

31. Required Tests/Data Derm assessment and Instrumentation Criteria Grading on Erythema Scale (0-none to 4-severe) Chromameter a-value, Mexameter Erythema Index, Digital Photography (before and after) Claim:Reduces skin redness

32. Suggested Tests/Data The product contains ingredients with proven anti-bacterial activity against p.acnes (propionibacterium acnes) Criteria Effective dosage levels of anti-bacterial agent, e.g. Salicylic Acid 0.5 -2.0%, Triclosan 0.1%. A 10% solution of the product should be able to inhibit p. acnes. Claim:Anti-bacterial

33. Oil Control Test Subjects: • Female teens, usually 13-19 yrs old • In good health, no allergy to cosmetics • Normal to Oily skin

34. Oil Control Test Design: • Base size of 20 to 30 • Double-Blind, Randomized • Control – vehicle and/or benchmark

35. Oil Control Test Site: • Forehead and/or naso-labial fold Product Usage: • Normal or habitual use • Usually 2x daily

36. Oil Control Before the study: • No leave-on products 24 hours before • Face must be cleansed 4 hours before • No caffeinated drinks 1 hour before • 30-minute acclimatization to room conditions

37. Oil Control Instrumental Evaluations: • Baseline, 1, 2, 4, 8 weeks • Sebumeter® – sebum level (g/cm2) • Sebufix® – pixels of sebum • Photography – shine/glare on the face

38. Oil Control Success Criteria: • Reduction in sebum vs. baseline • Higher reduction vs. vehicle • Higher reduction vs. benchmark

39. Oil Control Other Considerations: • Inclusion of safety assessment e.g. erythema, edema, dryness, scaling

40. Suggested Tests/Data Sebum Reduction Sebutape™ Sebumeter    Self assessment (150-based consumer research) Lipase Analysis via HPLC Criteria Chromatographic analysis of sebum (baseline, and at 2, 4, 6 and 8hrs). Skin sebum levels Statistically significant data vs baseline/control/ benchmark  A statistical significant reduction in triglycerides and fatty acids. Claim:Oil Control

41. SuggestedTests/Data Self assessment (150-based consumer research) Sebutape, Sebumeter Criteria Statistically significant results vs baseline/control/ benchmark The % change in sebum after washing with water versus washing with test product. Claim:Deep cleanses to remove oil and impurities

42. Claims related to Sunscreen Products

43. Waterproof Protects against skin burning UVB rays Protects against skin aging UVA rays Fights damaging free radicals Oil-Free Recommended by dermatologists Non-comedogenic (won’t clog your pores) SPF 45 Sweatproof

44. Required Tests/Data US FDA Sun Protection Factor Method (Australia uses their own method.) Criteria 20 person test. Males and Females. Fair skinned – Type I, II, and III. Claim: Sun Protection Factor SPF 15, 30, 45

45. Required Tests/Data Spectrophotometric Analysis (300-400nm) Criteria The product must not transmit more than 10% UV radiation at any wavelength between 320-360nm. Claims: Broad Spectrum Protection, Protects against skin-aging UVA rays, Protects against skin-burning UVB rays.

46. Required Tests/Data US FDA Immersion Test Criteria SPF rating after the prescribed immersion time. Claim: Waterproof for “x” hours

47. Suggested Tests/Data This type of claim relates to specific ingredients with anti-oxidant functions. In-vitro antioxidant, anti-ozone, anti-pollution activity Criteria An effective concentration of Vitamin E, C, AA2G, BHT, Lycopene etc. No changes in skin markers with product when exposed to pollutants Claims:Protects against / fights damaging free radicals

48. FAIRNESS

49. Lightening Test Subjects: • Female adults, usually 25-55 yrs old • In good health, no allergy to cosmetics • Preferably with slight to moderate dark skin tone

50. Lightening Test Design: • Base size of 30 to 50 • Double-Blind, Randomized • Control – vehicle and/or benchmark