No-Tox Botox: Natural Anti-Aging Ingredients Explained Moderator: Jessica Rubino, Senior Associate Editor, Delicious Living Dr. Linda Miles, L. Ac. DOM, Derma e Bodycare Chris Fields, VP Technology & Science NutriCosmetics /AFS Hyperpigmentation  Collagen/Elastin  Fine lines/Wrinkles  Inflammation Overall skin health  It will include a discussion of the latest science for cutting-edge topicals, collagen, and sunscreen

No-Tox Botox: Natural Anti-Aging Ingredients Explained Hyperpigmentation   Inflammation Overall skin health  It will include a discussion of the latest science for cutting-edge topicals, collagen, and sunscreen

PhotoDamage – Sun Protection Reduced Cell Turnover Cells remain on the surface longer Cellular mitosis creates abnormal keratinocytes Epidermal structure becomes disorganized Melanocytes over-produce melanosomes Hyperpigmentation Damaged collagen and elastin weakens dermal connective tissue Elasticity degradation Stratum Corneum: thick and dehydrated Skin cells are Misshaped & disorganized Excess melanin Causing uneven Skin tone Collagen & elastin fibers become weak & damaged

Natural Solutions Hydrating Anti-Inflammatory Antioxidants Depigmenting Olive Oil Chamomile Vitamin A, C, E Licorice Colloidal Oatmeal Coffee Extract Mulberry Hyaluronic Acid Feverfew Green / Black Tea Extract Arbutin Melon Extract Zinc Pomegrate Soy Krill Oil Fish Oil Grape Seed Extract Aloesin Sea Buckhorn COQ10 Acai Berry Extract Linoleic Acid Ceramides Vitamin C Resveratrol Niacinamide Natural Solutions

Inflammation Inflammatory skin diseases are the most common problem in dermatology. They come in many forms, from occasional rashes accompanied by skin itching and redness, to chronic conditions such as dermatitis (eczema), rosacea, seborrheic dermatitis, and psoriasis. Skin inflammation can be characterized as acute or chronic. Acute inflammation can result from exposure to UV radiation (UVR), ionizing radiation, allergens, or to contact with chemical irritants (soaps, hair dyes, etc.). This type of inflammation is typically resolved within 1 to 2 weeks with little accompanying tissue destruction. In contrast, chronic inflammation results from a sustained immune cell mediated inflammatory response within the skin itself. This inflammation is long lasting and can cause significant and serious tissue destruction.  Inflammatory skin conditions affect over 35 million Americans who annually spend over $2 billion to treat their symptoms.  The process of skin inflammation is complex and is still not completely understood. When the skin is exposed to a "triggering" stimulus, such as UV radiation, an irritant (e.g. soaps or fragrances), or to allergens, the cells in the skin produce a variety of inflammatory "hormones" called cytokines and chemokines. These "inflammatory messengers" bind to specific receptors on target cells and stimulate the production of additional inflammatory signaling "hormones". Some of these cause vasodilation while others activate nerve cells. Still other cytokines cause immune cells to leave the blood and migrate into the skin where they then produce more inflammatory hormones, as well as enzymes, free radicals, and chemicals that damage the skin. The end result of the initial triggering event is the amplification of a large inflammatory response that, while designed to help the skin fight infection from invading bacteria, actually causes considerable damage to the skin

Avenanthramide (ug/ml) - MOA And although typically ingested, oats have been shown to work topically. British researchers at the royal London hospital said topical formulations of natural colloidal oatmeal should be considered an important component of therapy for atopic dermatitis and other conditions, noting it may allow for reduced use of corticosteroids and calcineurin inhibitors.12 Colloidal oatmeal has an extensive history of beneficial use in dermatology. In recent years, in vitro and in vivo studies have begun to elucidate the multiple mechanisms of action of naturally derived colloidal oatmeal. Evidence now describes its molecular mechanisms of anti-inflammatory and antihistaminic activity. The Avenanthramide, a recently described component of whole oat grain, are responsible for many of these effects. Studies have demonstrated that Avenanthramide can inhibit the activity of nuclear factor kappaB and the release of proinflammatory cytokines and histamine, well known key mechanisms in the pathophysiology of inflammatory dermatoses. Cerio r et al. “Mechanism of action and clinical benefits of colloidal oatmeal for dermatologic practice” J Drugs Dermatol. 2010 Sep;9(9):1116-20 Agents with adjunctive therapeutic potential in atopic dermatitis Wallo et al, 65th Annual AAD, Feb 2006, Washington DC

Avenanthramides- Clinical Evidence Oat Avenanthramides: new actives to reduce itch sensation in skin. Schmaus Et al, 23rd Congress of Intr Fed of Soc Cos Chemists, Oct 2006 Orlando FL

Mediator Release (TNF-α ICD50 (ug/ml) Feverfew Compound Mediator Release (TNF-α ICD50 (ug/ml) Feverfew PFE 0.13 Green Tea 4.6 Echinacea 20 Boswelia seratta 24.5 Licorice Extract 54.7 Black tea 79.9 White Tea 125 Bisabolol 160 Chamomile 320 Aloe Vera 535 Olive Leaf 675 PFE - purified feverfew extract TNF-α ICD50 = conc req to achieve 50% inhibition of tumor necrosis factor-α Johson & Johnson, 2006 Tierney et al – 63rd Annual Meeting of the AAD – inhibition of UV Radiation induced erythema Liebel et al, ADD Feb 2005 – demonstrated strong anti-inflammatory properties Nebus et al ADD Feb 2005 – Significant improvement in erythema (n=31)

Glycyrrhiza- Licorice Anti-bacteria Anti-protozoan Anti-tumor Depigmentation ANTI-INFLAMMATORY: Razor Test Shaved Skin UV Induced Erythema Test Hemoglobin reflectance Cytokine production Focus on Erythema, measure skin redness of shaven skin using reflectance spectroscopy Anti-inflammatory efficacy of Licochalcone A Kolbe et al Arch Dermatol Res (2008)

Hyperpigmentation Two Forms of Melanin produced in the Epidermis: Pheomelanin Eumelanin Melanin Biosynthesis (melanogenesis) Genetics Environmental Lifestyle Medications Production Action of the Enzyme Tyrosinase Copper Oxygen Sulfur Hyper Pigmentation is a condition where the body produces too much melanin thereby causing to become darker than usual. Hyper pigmentation can occur due to excessive sun bathing or drug reactions. Many a time wounds and scars leave a darker patch of skin. Birthmarks, moles, and aging spots are also indications of hyper pigmentation. It is important to keep on the alert for any change in size, color or texture for indications of skin cancer.
 Melanin biosynthesis (melanogenesis) is influenced by genetics, environmetal factors, diet and medication. The production of melanin by specialized cells called melanocytes (in the basal layer of the epidermis in light skinned people and in the basal as well as horney layer in dark skinned people) occures through the action of the enzyme tyrosinaase. The rate-limited step in melanogenesis is the conversaion of L0tyrosinase to melanin, through the action of tyrosinase. Copper and oxygen act as catalysts. Other enzymes also control melanin produciton, particularlry in the presense of sulfur.

Melanin Production Pathways

Transcription of the tyrosinase Gene Melanin Production Tyrosinase Activated Tyrosinase Melanin Transcription of the tyrosinase Gene Pigmentation pathway: Tyrosinase Inhibition Inhibition of melanin/melanos ome transport into keratinocytes Increased Epidermal turnover Glycosylation + sugar Signals: UV Irritation Hormones Radicals Tyrosine Varioation in skin pigmentation attributes to the levels of melanin produced and the number of melanocytes present. Melanin granules synthesized in the melanocytes are then transferred from the cytoplasm of the melanocytes to the basal cytoplasm of the keratinocytes – Forming a protective covering in the inner layers of the epidermis, absorbing UV rays and inhibiting their penetration. Melanin is packed into melanosomes Keratinocytes

Melanin Production Pigmentation pathway: Tyrosinase Inhibition Glycation Inhibitors: N-acetyl glucosamine Pigmentation pathway: Tyrosinase Inhibition Inhibition of melanin/melanos ome transport into keratinocytes Increased Epidermal turnover Gene production inhibitors: retinoids Tyrosinase Activated Tyrosinane Melanin Transcription of the tyrosinase Gene Signal Reduction: Sunscreen, AA Glycosylation + sugar Signals: UV Irritation Hormones Radicals Tyrosine Activity Inhibitors: Kojic Acid Arbutin Licorice Varioation in skin pigmentation attributes to the levels of melanin produced and the number of melanocytes present. Melanin granules synthesized in the melanocytes are then transferred from the cytoplasm of the melanocytes to the basal cytoplasm of the keratinocytes – Forming a protective covering in the inner layers of the epidermis, absorbing UV rays and inhibiting their penetration. Melanin is packed into melanosomes Melanosome Transfer Inhibition: Niacinamide, Soy Protein Keratinocytes

A review of the history Mechanism Ingredient Activity UV Absorption Titanium dioxide, oxybenzone Blocks UV Tyrosinase synthesis inhibitor Glucosamine, galactosamine Exhibits cytotoxic effects Tyronsinase Inhibitor Ascorbic Acid, kojic acid, mulberry extract, licorice, arbutin, Curcuminoids Melanin Biosynthesis Kojic acid Chelates copper ions Cytotoxic effects to melanocytes Hydroquinone Safety Issues Reduce preformed melanin Tocopherols, ascorbic acid Inhibits polymerization of melanin Endothelin-1 inhibitor Chamomilla extract Erigeron breviscapus Inhibits ET-1 induced DNA synthesis Endothelin inhibitors are reported to work faster than tyronsinase inhibitors because their MOA is outside the melanocyte cell membrane. (Tyrosinase inhibitors have to crsoss four barriers for deactviation (stratum corneum, epidermis, melanocyte membrane and the melanosome membrane. To discover an active skin depigmenting agent, we isolated a novel inhibitor of melanin biosynthesis from the methanol extract of Erigeron breviscapus using a bioactivity-guided fractionation and identified it as (2Z,8Z)-matricaria acid methyl ester by means of spectroscopic analysis. The compound showed strong whitening activity in melan-a cell. Compared with arbutin (IC(50)=4.0 mM) as a positive control, the depigmentation IC(50) value for (2Z,8Z)-matricaria acid methyl ester was 25.4 muM in B16F10 melanoma cell. Moreover, its inhibitory effect on tyrosinase, the key enzyme of melanogenesis, was examined by in vivo and in vitro tyrosinase assay and Western blot. The results indicate that (2Z,8Z)-matricaria acid methyl ester isolated from Erigeron breviscapus is a promising compound that could be useful for treating hyper-pigmentation as skin-whitening agents.

Active Soy Performance Efficacy of Soy Moisturizer in PhotoAging, Wallo et al, JDD, Sept 2007

Erigeron breviscapus Depigmentation of Melanocytes by Matricaria Acid Methyl Ester isolated from Erigeron breviscapus. Luo et al, Biol Pharm Bull 32(6) 2009

Combination Therapy Niacinamide & N-acetyl glucosamine Reduction in the appearance of facial hyperpigmentation after use of moisturizer with combination Of topical niacinamide and N-acetyl glucosamine: results of a randomized, double-blind study Kimball et al., B J Dermatol, 2010, 162(2)

Combination Therapy Niacinamide & N-acetyl glucosamine Reduction in the appearance of facial hyperpigmentation after use of moisturizer with combination Of topical niacinamide and N-acetyl glucosamine: results of a randomized, double-blind study Kimball et al., B J Dermatol, 2010, 162(2)

Combination Therapy Niacinamide & N-acetyl glucosamine Dual Sites of Activation N-acetyl glucosamine – Inhibits Tyrosinase Niacinamide – Inhibits the transfer of melanin- containing melanosomes to the outer layers.

Inflammation+Hyperpigmentation: Combination therapies Dr. Raval, MD Raval Facial Aesthetics, Rocky Mountain Laser Clinic, 2011 CDG = Glucarate 3 w/w% GCA = Green Coffee Extract (50% CA) 3 w/w% Combination CDG+GCA (3 w/w%)

Sunscreen Advances UVA / UVB Absorption Zinc Oxide / Titanium Dioxide Photostabilizers – Octocrylene Avobenzone Ecamsule Antioxidants – Reduce ROS Green Tea Soy Vitamin C

Pisum Sativum Extract Pisum Sativum Extract for Safe & Self-tanning, Imbert et al Cosmetics & Toiletries August, 2009 V124.