© mibelle biochemistry, switzerland 2007 detoxophane activating the skin‘s detoxification system

© Mibelle Biochemistry, Switzerland 2007

DetoxophaneActivating the skin‘s detoxification system


Mom was right: „Eat your vegetables, they are good for you!“

• Phytonutrients are plant chemicals reported to exert a specific effect on health or on amelioration of diseases

• Health benefits of vegetables: Lower risk for certain cancers, stroke, and high blood pressure


The Vegetables of the Mustard Family

• Broccoli sprouts are now extreme popular as food supplements in the USA

• Vegetables of the mustard (Brassicaceae) family have been shown to be extremely healthy

• They contain a high concentration of sulforaphane

• Members of the Brassicaceae vegetables are broccoli, cauliflower, radishe, water and garden cress


Garden Cress Sprouts: The Source for a New Cosmetic Ingredient

• Garden cress is a suitable form for sulforaphane for cosmetics

• Fresh sprouts are homogenized and extracted with water

• The extract is purified, concentrated and encapsulated into liposomes


Composition

• Lepidium Sativum Sprout Extract• Glycerin• Lecithin• Phenoxyethanol• Aqua/Water


Isothiocyanates: Molecular Structures

In the plant, isothiocyanatesare present as glucosederivatives, called glucosinolates.When the plant cells are broken,the enzyme myrosinase isliberated. This thioglucosidasetransforms glucosinolates intofree glucose and isothiocyanates. Sulforaphan

e S

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H3C N C S


Sulforaphane: Mechanism of Action

Sulforaphane activates the body‘s detoxification system composed of phase I and phase II enzymes:

Phase I reactions increase the reactivity of hydrophobic molecules by addition of functional polar groups (->reactive oxygen species). If these reactive molecules are not further metabolized by phase II conjugation, they may cause damage to DNA and induce mutations.

Phase II reactions are either conjugation reactions of phase I products with small hydrophilic substances to form water-soluble products that can be excreted or they are antioxidant enzyme reactions that inhibit strong oxidant formation.


Detoxophane is More than an Antioxidant

UV

Toxin

AGE

DNA damage

Lipidperoxides

ROSAntioxidant Detoxophane


Activation of Gen Expression with the Antioxidant Response Element (ARE)

• The antioxidant response element ARE is a regulatory promoter sequence of genes coding for antioxidant systems and phase II detoxification enzymes

• The transcription factor Nrf2 is the critical regulator of ARE-dependent gen expression

• Under normal conditions Nrf2 is blocked by the repressor Keap1• Sulforaphane was found to disrupt the Nrf2/ Keap1 complex


Detoxophane: The Next Generation Antioxidant

Stimulates the expression of antioxidants and detoxification enzymes in the skin to protect against

• Environmental pollutants (cigarette smoke, exhaust fumes, smog, endotoxins, heavy metals)

• Reactive oxygen species (extrinsic and intrinsic)

• DNA damage and thus premature skin aging


Modulation of Enzyme Expression

Method

• Human keratinocytes were grown to 80% confluence in a standard growth medium.

• Then cells were incubated during 24 hours with different concentrations of Detoxophane.

• Cells were harvested and total RNA extracted.

• Real time quantitative PCR was used to analyze gene expression

Phase I - Cytochrome P450 isoenzyme 2E1 (CYP2E1)

Phase II - NADPH: quinone reductase 1 (NQO1)

- Heme oxygenase 1 (HO-1)

- Thioredoxin reductase 1 (TrxR1)


Modulation of Enzyme Expression

Enzyme expression rel. to control (%)

Concentration of Detoxophane 0.05% 0.2%

Phase I enzyme

Cytochrome P450 isoenzyme 2E1 -40 -38

Phase II enzyme

NADPH:quinone reductase 1 75 214

Heme oxygenase 1 212 4182

Thioredoxin reductase 1 184 2316S-131


Protection Against Oxidative Stressors

Method• Growth of human keratinocytes to 80% confluence in a standard growth medium• Pre-treatment for 24 hours with 0.05% Detoxophane• Incubation for 4 hours with increasing concentrations of toxins:

- 4-hydroxynonenal HNE

- tert-butyl hydroperoxide t-BH • Analysis of cell viability with MTT assay


Protection Against Tert-Butyl Hydroperoxide

S-148


• Detoxophane exerted an impressive protection against t-BH. At 0.05% Detoxophane, the IC50 value increased from 258 microM of the control to 646 microM.

• Detoxophane exerted a moderate protection against HNE. At 0.05% Detoxophane, the IC50 value increased from 76 microM of the control to 90 microM.

Protection Against Tert-Butyl Hydroperoxide


Summary

• Vegetables of the mustard family,

such as broccoli and garden cress

sprouts, are known for their

remarkable health benefits• The phytonutrient sulforaphane

has been identified as the active

compound• Garden cress sprouts have been

used to develop Detoxophane as

cosmetic sulforaphane preparation


Claims

Detoxophane…• Activates the detoxification

system of skin cells• Protects skin against

pollutants • Prevents visible signs of skin

aging• Protects against city pollution • Prevents DNA damage• Reveals a healthy looking

skin


Application

• Anti-Ageing Formulations • Detox City Creams • Sun Protection Formulations • Hand Creams• Protective Foundations


Film

© mibelle biochemistry, switzerland 2007 detoxophane activating the skin‘s detoxification system

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