Revolutionary and New Category in Skin Health 2019 Micah Chemistry - PowerPoint PPT Presentation

Revolutionary and New Category in Skin Health 2019

Micah™ Chemistry

ante ante- a prefix meaning “before,” of Latin origin

Micah ante oxidant™ Micah anteoxidant TM - to prevent the visible signs of photoaging, of Hallstar origin

Micah ante - oxidative stress - inflammatory - DNA damage - pollution - blue light - UVA damage - wrinkles - collagen degradation - fine lines - aging

Micah ante oxidant™ CLINICALLY PROVEN TO Prevent 100% of UVA induced oxidative stress and DNA damage Prevent 55% of UVA induced inflammatory cytokines (IL-6) Prevent 35% of UVA induced matrix degrading enzymes (MMP-1) Prevent 31% of Visible Light induced oxidative stress

HALLSTAR’S VISION: PROTECT CONSUMERS FROM THE ELEMENTS UVB | UVA-II | UVA-I | VISIBLE LIGHT | IR | POLLUTION WORK IN CONCERT TO PROTECT CONSUMERS

CONSUMER CHOICES: EUROPE

CONSUMER CHOICES: EUROPE 4x Increase in products launched with Blue Light claims since 2016 5,413 Number of products launched 6,571 with Anti-Pollution claims in EMEA since 2014 Number of products launched with UVA claims in EMEA since 2014

THE MICAH COMPOUND & MICAH COMPOUND & MECHANISM OF ACTION SAFETY DOSSIER

Micah: Compound Micah Compound: Description n-Butyl Ester ▪ Fused Ring Cyanoacrylate ▪ Molecular Weight: 454 g/mol ▪ Proposed INCI: Polycyclic Di-Ester (Official name expected Feb 2019) ▪ At projected Use Level in Cosmetic Lotion ≤ 0,75%: ► SPF negligible ► UVA absorbance negligible (~0.00001) ► No Visible Light absorption (at any use level) ▪ CAS: 17

Micah Compound: Safety & Toxicology SAFETY DOSSIER ✓ Genotoxicity/Ames test ✓ Ocular irritation (HET-CAM) ✓ Phototoxicity Clinical – in vivo (22 volunteers) ✓ Phototoxicity 3D Skin Model – in vitro ✓ Phototoxicity Human Skin Explant - ex vivo ✓ Sensitization/HRIPT (50 volunteers) 18

THE MICAH COMPOUND & MECHANISM OF ACTION MECHANISM OF ACTION

UV & VISIBLE LIGHT: PENETRATION INTO HUMAN SKIN

UVA / Visible Light- induced ROS/FR Generation: ‘Base Case’ .- 1 O 2 O 2 H 2 O 2 ● OH Singlet Oxygen Pathway MAPK Oxidation 3 2 UVA / Vis 3 O 2 ▲ ▲ 8-OH-dG AP-1 ▲ ▼ TGF- b DNA Damage PS* ▲ Molecular Oxygen Lipid Damage ▲ Protein Damage Photoexcited MMP-1, IL6, IL1 a ▲ Chromophore 1 ▲ Collagen I, III (V) Breakdown ▼ Procollagen Production PS e.g. Porphyrins, Flavins VISIBLE SIGNS OF PHOTOAGING 1 Endogenous photosensitizers (PS) absorb solar radiation – inducing of photoexcited states (PS*) 2 Excited photosensitizer (PS*) transfers energy to 3 O 2 to generate 1 O 2 and subsequent ROS / FR 3 ROS / FR can lead to DNA, lipid, protein damage and mediate production of MMPs, IL’s: PHOTOAGED SKIN

UVA / Visible Light-induced ROS/FR Generation: Sunscreens & Antioxidants  ANTIOXIDANTS .- 1 O 2 O 2 H 2 O 2 ● OH Singlet Oxygen Pathway MAPK Oxidation UVA / Vis  SUNSCREENS 3 O 2 ▲ ▲ 8-OH-dG AP-1 ▲ ▼ TGF- b DNA Damage PS* ▲ Molecular Oxygen Lipid Damage ▲ Protein Damage MMP-1, IL6, IL1 a ▲ ▲ Collagen I, III (V) Breakdown ▼ Procollagen Production PS e.g. Porphyrins, Flavins VISIBLE SIGNS OF PHOTOAGING  SUNSCREENS: Specific sunscreens absorb and/or reflect UVA, but performance diminish in the UVA1 portion of the solar light spectrum. Sunscreens may ‘color’ in the near -Visible portion of the spectrum.  ANTIOXIDANTS: Antioxidants seek to neutralize ROS / FR . . . but only after the ROS/FR cascade has started.

UVA / Visible Light-induced ROS/FR Generation: Micah Chemistry .- 1 O 2 O 2 H 2 O 2 ● OH Micah Compound Singlet Oxygen Pathway MAPK Oxidation UVA / Vis 3 O 2 ▲ ▲ AP-1 8-OH-dG ▲ ▼ TGF- b DNA Damage PS* ▲ Molecular Oxygen Lipid Damage ▲ Protein Damage MMP-1, IL6, IL1 a ▲ ▲ Collagen I, III (V) Breakdown ▼ Procollagen Production PS e.g. Porphyrins, Flavins UV/Vis PHOTOAGING SURPRESSED ▪ Micah quenches excited states of endogenous photosensitizers (PS*), suppressing the generation of singlet oxygen ( 1 O 2 ) and subsequent ROS / FR. ▪ Micah stabilizes by an electron transfer mechanism, and therefore will not impart color onto skin.

UVA / Visible Light-induced ROS/FR Generation: Micah Chemistry “ It is of the .- 1 O 2 O 2 H 2 O 2 ● OH Micah Compound Singlet Oxygen highest Pathway MAPK Oxidation UVA / Vis importance to 3 O 2 ▲ ▲ AP-1 8-OH-dG ▲ ▼ TGF- b DNA Damage PS* effectively fight ▲ Molecular Oxygen Lipid Damage ▲ Protein Damage MMP-1, IL6, IL1 a ▲ Singlet Oxygen” – Paolo Giacomoni, former R&D Director, ▲ Collagen I, III (V) Breakdown ▼ Procollagen Production Estee Lauder & L’Oreal PS e.g. Porphyrins, Flavins UV/Vis PHOTOAGING SURPRESSED

UVA / Visible Light-induced ROS/FR Generation: Singlet Oxygen 1 O 2 ▪ Singlet Oxygen reacts with proteins, lipids and nucleic acids. ▪ Outside Cell: Lipid peroxidation ▪ Inside Cell: Pro-inflammatory cytokines and MMP-1 • ▼ Skin elasticity • ▼ Skin resilience ▪ Damage to cell DNA; 8 OH-dG lesion 8 OH-dG Micah suppresses the generation of Singlet Oxygen

Singlet Oxygen and ROS Prevention: in vitro • HaCaT cells pretreated with aminolevulinic acid (ALA), a strong photosensitizer of singlet oxygen - 1 O 2 • HaCaT cells fluoresce when irradiated at 630nm • HaCaT cells pretreated with ALA and Micah show no fluorescence, indicating Micah quenched the electron transfer cascade that produces singlet oxygen 1 O 2 and other ROS. Visible Light (-) Micah (-) Visible Light (+) Micah (-) Visible Light (+) Micah (+) Cells pre-treated with ALA Cells pre-treated with ALA Cells pre-treated with ALA A B C Protocol: The HaCat cells were seeded in the 60mm cell culture dishes, after the cells became confluent, the cells were treated with 8mM ALA for 2 hours, and then different concentration of Rx14401, from 0 (no Rx14401 control) to 2.0mM, were added to the cells. After 1 hour, each plate of cells was irradiated at 630 nm for 3 minutes (for no irradiation control, the wells are wrapped with aluminum foil). After washing with PBS buffer, the cells were stained with DFFDA for 30 minutes. The cells were washed with PBS 3 times again and lysated by sonication. The cell lysates were analyzed with fluorescent plate reader to access the cellular ROS.

THE MICAH COMPOUND PROOF OF CONCEPT: ROS / FREE RADICAL PREVENTION – VISIBLE LIGHT & ex vivo and in vivo MECHANISM OF ACTION Gematria Labs, Berlin, Germany Dr. Schrader Institute, Germany

Micah effectively prevented visible- light induced ROS / FR and outperformed 1% Vitamin E.

VISIBLE LIGHT CLINICAL STUDY: DESIGN SUBJECTS FORMULATIONS LIGHT SOURCE & RESPONSES ▪ ▪ Chemiluminescence – photon 10 Subjects 1. Vehicle emission counts ▪ Skin Types I - III 2. Micah 0,2% ▪ Visible Light: 400 – 700 nm 3. Micah 0,75% 2 m l / cm 2 applied 2x per day (AM / PM) for 4 days Visible Light (400 – 700 nm) Energy released from oxidative stress in form of photon. AM Photomultiplier tube (PMT) Day Day Day Day continuous monitoring and 1 2 3 4 measurement of photon PM energy.

VISIBLE LIGHT CLINICAL STUDY: RESULTS At a use level of 0,75%, Micah demonstrated a 31% reduction in visible light-induced oxidative stress. Further optimization possible with a higher does of Micah. ▼ 31% No statistical difference

THE MICAH COMPOUND PROOF OF CONCEPT: & UVA: IL6, 8-OHdG, MMP-1 in vivo / clinical studies on human patients MECHANISM OF ACTION Skin Investigation and Technology Hamburg, Germany

UVA CLINICAL STUDY: DESIGN SUBJECTS FORMULATIONS RESPONSES ▪ ▪ 10 Subjects 1. Untreated Skin DNA Damage (8-OH-dG) ▪ ▪ Skin Types I - III 2. Untreated Skin / Irradiated IL-6 (ELISA) ▪ ▪ Age; 30.0 +/- 8.2 years 3. Micah 0,2% / Irradiated MMP-1 (ELISA) 4. Micah 0,5% / Irradiated 2 m l / cm 2 applied 2x per day (AM / PM) for 14 days 20 J / cm 2 UVA (320 – 400 nm) AM Day Day Day Day Day Day Day Day Day Day Day Day Day Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 PM t = 0 hrs and t = 24 hrs

UVA CLINICAL STUDY: DNA DAMAGE – 8-OH-dG At use levels between 0,2 and 0,5%, Micah significantly reduced the presence of UVA-induced 8- OH-dG, a maker associated with photoaged skin. 8-OH-dG levels returned to baseline with Micah.

UVA CLINICAL STUDY: DNA DAMAGE – 8-OH-dG High intensity

UVA CLINICAL STUDY: MMP-1 At a use level of 0,5%, Micah demonstrated a 35% reduction in the presence of MMP-1, a marker associated with the breakdown of the extracellular matrix. ▼ 35% No statistical difference

UVA CLINICAL STUDY: IL-6 At a use level of 0,5%, Micah demonstrated a 55% reduction in the presence of IL-6, a marker associated with skin inflammation, redness, irritation and barrier function. ▼ 55%

THE MICAH COMPOUND & PROOF OF CONCEPT: ROS / FREE RADICAL PREVENTION, IL1 a , IL6, 8-OHdG, MMP-1 MECHANISM OF ACTION ex vivo human skin BEN GURION UNIVERSITY, ISRAEL

 Positive Controls → * Significantly different from non-irradiated controls # significantly different from irradiated vehicle

 Positive Controls → * Significantly different from non-irradiated controls # significantly different from irradiated vehicle