Actives

HerbaProtect ® NOX

New The Night-Time Sun Care Active Data • Reduction of light-mediated oxidative stress • Prevention of delayed UV-induced skin damage • Protection against photoaging • Deceleration of premature aging

At a Glance

The main approach to protect against sun-induced skin damage is the prevention or reduction of sun exposure. Certainly, stepping out of the sun at the right time can minimize the negative consequences significantly. However, skin damage not only occurs immediately while being in the sun. New discoveries indicate that the skin stress mediated by sunlight continues for hours after the skin has been exposed to UV irradiation; it continues even at night-time. Sunlight induces the generation of oxygen and nitrogen radicals that bring the skin’s own melanin to an excited state. Subsequently, the melanin transfers this excess energy to the DNA, which ultimately causes delayed skin damage, so-called ‘dark CPDs‘ [1] . Besides sun-induced oxidative stress, free radicals occur in response to a variety of intrinsic and extrinsic factors and promote premature aging.

HerbaProtect NOX is a COSMOS-approved innovative sun care and anti-aging active that protects against delayed sun-induced skin damage and premature aging. Three active components, namely plant extracts from pomegranate flower, perilla leaf and kakadu plum, have been combined in a preservative-free, standardized and concentrated glycerol-based solvent system.

HerbaProtect NOX acts on reactive oxygen and nitrogen species as well as on peroxynitrite, which is the main driver of delayed sun-induced skin damage. In vitro and in vivo testing clearly demonstrated an immediate and long-term improvement of the skin’s antioxidative capacity. An increased defence status of the skin against sun-induced oxidative stress eventually reduces sun-induced skin damage and premature aging.

Taken together, HerbaProtect NOX is a perfect addition for sun care, after-sun or anti-aging formulations that aim at protecting the skin against delayed sun-induced skin stress mediated by UV radiation, visible light or even infrared radiation.

© Lipoid Kosmetik AG Sun Damage Continues after Sunset

The need for ‘night-time sun care products‘ Exposure to UV radiation is the major cause of physiological skin damage, such as photoaging and erythema. Recent research, however, has shown that UV-induced skin damage can occur several hours after the radiation and even at night-time. Alarmingly, it appears that half of the damage caused by sunlight actually occurs long after exposure to the sun [1] .

It is actually not surprising that sun exposure not only induces immediate high-level skin damage, but also sustained low-level skin damage. This fact becomes obvious when considering the natural tanning reaction, which is the skin’s response to damage caused by UV radiation: Immediate pigment darkening begins during UV exposure and is instantaneously visible, as it is caused by melanin already present in the skin. The delayed tanning reaction arises in the days following UV exposure and is stimulated by endogenous signalling reactions. This example shall help to understand that stepping out of the sun is not sufficient to protect against its damage. In fact, the sun can initiate a persistent challenge to the skin, referred to as delayed sun-induced skin damage (Fig. 1, 3), which needs to be additionally counteracted.

This rather disregarded, delayed part of the sun-induced skin damage illustrates that novel and innovative cosmetic concepts are required to combat sun damage entirely, for instance ‘sun protection after sunset’ or ‘night-time sun care products’.

Fig. 1: Sun protection after sunset. Sunlight induces a cascade of signals that occur even under darkness and eventually lead to delayed cellular damage. This implies that mere sun protection is not enough and that ‘night-time sun care products’ that provide more than moisturizing properties are requi- red after sun exposure, in order to prevent the ongoing damage of UV radiation.

We make beauty natural. Counteracting Delayed Sun-Induced Skin Damage

The generation of peroxynitrite causes DNA damage long after UV exposure UV radiation generates oxidative stress and DNA damage almost instantaneously, i.e. within picoseconds upon sun exposure. However, current research suggests that sun-induced damage continues for several hours after UV exposure [1] . The explana- tion for this intriguing discovery is as follows:

• Sun exposure induces the enzymes NADPH oxidase (NOX) and nitric oxide synthase (NOS), which generate reactive oxy- gen and nitrogen species (ROS and NO, respectively) in the skin. The reaction of these molecules with each other leads to the formation of peroxynitrite (ONOOˉ). • The highly reactive peroxynitrite, in turn, reacts with melanin resulting in excited melanin, which slowly diffuses into the nucleus. In the nucleus, the excited melanin either emits light, or transfers its excess energy to DNA, eventually creating DNA damage, such as cyclobutane pyrimidine dimers (CPDs). Since the formation of these CPDs occurs hours after the original UV exposure, they are referred to as ‘dark CPDs’ [1,2] . This delayed reaction apparently causes the same amount of DNA damage as immediate effects. The slow course of the re- action, however, gives time for intervention. Indeed, antioxidant cosmetic active ingredients that scavenge free radicals and nitric oxide production or that quench excited melanin represent a promising approach to counteract delayed UV-induced skin damage.

Increasing the skin’s antioxidant capacity – a universal photoprotection and anti-aging strategy Skin aging is mainly attributed to UV radiation, but it has become evident that other parts of the solar spectrum, particularly infrared-A (IR-A) or even visible light, also play a role in mediating aging (Fig. 2). Effects of IR radiation and visible light are comparable with the action of UV radiation as they also cause skin stress by initiating reactive oxygen species (ROS). Because ROS constitutes the basis of any solar-induced damage, the antioxidant status of the skin is fundamental. Currently, there is wide consensus that topical application of antioxidants represents the most effective approach to provide protection against IR- or visible light-induced ROS generation in the skin. Thus, powerful antioxidants are considered a fundament of photopro- tection products [3-6] .

Indeed, HerbaProtect NOX is a very powerful antioxidant (Fig. 6), minimizes H 2O2- and peroxynitrite-induced cell stress (Fig. 7) and increases the antioxidant capacity of the skin for several hours (Fig. 10). These properties make HerbaProtect NOX a perfect ingredient for anti-aging and sun care products that aim at providing any kind of photoprotection, whether against UV, IR or visible light.

Content in Sunlight 5 % 50 % 45 %

ROS/RNS Generation, Photoaging

DNA Damage, DNA Oxidation Heat Sunburn

Blocked by Atmosphere

UV-C UV-B UV-A VisibleVi si bl e lightLightLi gh t Infrared

200 290 315 760 1000 nm

Fig. 2: The solar spectrum and its effects on skin. Only UV-B carries enough energy to damage DNA directly. UV-A is 20 times more abundant, but con- tains less energy and thus only damages DNA via indirect routes, i.e. formation of free radicals or peroxynitrite. IR radiation is even more abundant than UV light, but ‘only’ has enough power to generate reactive oxygen and nitrogen species (ROS, RNS) [7] . Nevertheless, the contribution of IR-A to ROS generation is around one fourth of that of UV radiation [8] .

We make beauty natural. Composition

Pomegranate flower ( Punica Granatum ) – ROS and NO scavenger Common names: Granatapfel (DE); pomegranate (EN); grenadier, grenadier commun (FR)

The name pomegranate is derived from the French words “pomme garnete” (= see- ded apple). Interestingly, every fruit seems to be composed of exactly 840 seeds. The exotic fruits, which originated in Iran over 4000 years ago, have been mentioned even in Greek legends and ancient texts, including the Bible. They became famous not only as an important food source, but also because of their medicinal purposes [9] . In some cultures, pomegranate fruits symbolize life after death, fertility, health and prosperity. It is, therefore, not surprising that skin care manufacturers nowadays include pomegranate extracts in their products.

Fruits, seeds and leaves are rich sources of valuable pomegranate phytochemicals, such as flavonoids and tannins. Moreover, these plant parts exhibit high antioxidant and photoprotective properties [10,11] . The flowers, however, have superior antioxi- dant activity [12] , possess the ability to scavenge reactive oxygen and nitrogen spe- cies (ROS, NO) [13] and to protect skin cells against UV-A radiation [14] .

Additionally, pomegranate flower extract stimulates the production of collagen and elastin, while decelerating their breakdown [15] . Hence, the use of pomegranate flower extract represents an efficient and appealing anti-aging strategy to attenu- ate aging- as well as sun-mediated undesirable skin changes.

Perilla leaf ( Perilla Frutescens ) – NOX and NOS suppression Common names: Schwarznessel, Sesamblatt (DE); beefsteak plant, perilla mint (EN); périlla (FR)

Perilla leaves are not only commonly used as decorative elements in Asia, but are also consumed as a vegetable or applied as herbal medicine. Leaf extracts display a range of biological activities, including anti-inflammatory and antioxidant proper- ties. Notably, Perilla leaf extract is suggested to be a potent agent for prevention of skin aging, as it has protective effects against UV-induced dermal matrix damage [16] . More pre- cisely, perilla leaf extracts attenuate the expression of nitric oxide synthase (NOS) resulting in lower levels of reactive nitrogen species (NO) [17] . Moreover, perilla leaf extracts seem to inhibit the function of NADPH oxidase (NOX) and reduce the levels of reactive oxygen species (ROS) that are produced by this enzyme [18] . The main active ingredients were identified as polyphenols, inclu- ding rosmarinic acid, a potent ROS scavenger, as well as luteolin and apigenin [18-21] .

Taken together, these properties substantiate that perilla leaf extract can help to attenuate peroxynititri- te formation and counteract delayed sun-induced cell damage.

We make beauty natural. Composition

Kakadu plum ( Terminalia Ferdinandiana ) – antioxidants against oxidative stress Common names: Buschpflaume (DE); gubinge, billygoat plum, murunga (EN) ; prune de kakadu (FR)

Kakadu plum is an Australian native plant food. The fruits have been eaten by Aborigines on long treks or hunting trips, and were considered more valuable as a medicine rather than a food. It regained popularity when it was rated as richest source for vitamin C of any plant in the world. The content of vitamin C varies between 3.5 and 5.5 % of the wet weight of the fruit, which is 100 times the concentration in oranges [22] . Due to its extraordinary high vitamin C levels, the primary use of the fruits is for health food, pharmaceuticals and personal care industries. Furthermore, Kakadu plum is increasingly found in innovative restaurant menus, in health shops, and cosmetics.

In addition, Kakadu plum contains many other valuable compounds such as gallic acid, ellagic acid or luteolin, which also contribute to its high antioxidant activity. The phytochemicals of the Kakadu fruit may reduce oxidative stress in several ways: either directly by scavenging of ROS or indirectly by inhibition of the NO- producing enzyme NOS [23-25] .

In summary, Kakadu plum, with its extraordinary high antioxidant levels, has the potential to decrease cellular oxidative stress and counteract immediate as well as delayed sun-induced skin stress.

We make beauty natural. Mode of Action

Stepping out of the sun is not sufficient to protect entirely against UV-induced skin stress as damage continues after sun exposure. Sunlight induces NOX (NADPH oxidase) and NOS (nitric oxide synthase), which causes the generation of reactive oxygen and nitrogen species (ROS and NO), followed by an increased peroxynitrite (ONOOˉ) formation [1,26] . Peroxynitrite is a detrimental oxidant because it is capable of exciting melanin. Excited melanin slowly discharges its energy, either by emitting light or by transferring the energy to DNA, which eventually causes delayed cellular damage, such as the formation of cyclobutane pyrimidine dimers (‘dark CPDs’) [1] . Hence, interference with peroxynitrite formation and melanin excitation provides a promising skin protection strategy and paves the way for innovative anti-aging, after-sun and light protection concepts. HerbaProtect NOX increases the skin’s antioxidant capacity, counteracts delayed sun-induced skin stress and reduces premature aging through several mechanisms:

• The active components of HerbaProtect NOX inhibit the enzymes NOX and NOS, and thereby reduce ROS and NO produc- tion and limit oxidative stress. • Moreover, HerbaProtect NOX has excellent antioxidative activity and scavenges peroxynitrite and ROS. • The efficient decrease of peroxynitrite and ROS levels results in a reduction of delayed UV-induced skin damage, such as ‘dark CPDs’. • In addition, an enhanced antioxidative capacity of the skin protects against damage mediated by the entire solar spec- trum (also visible light and infrared) and reduces premature aging.

Skin exposure to sunlight: 2EACTIONÏTOÏ56ÏWITHINÏTHEÏlRSTÏFEWÏMINUTES HerbaProtect NOX

No protection Epidermis

HerbaProtect NOX inhibits NOS and NOX

UV-induced melanin synthesis NOX Melanin NOX ROS ROS + - NOS NO - ONOO NO - NOS Peroxy- NOXNO X NOS nitrite ROS

ONOOONOO-

Molecular procession Scavenging of peroxynitrite and ROS up to 3h after sunlight exposure

Peroxynitrite- and melanin-mediated inductionn Reduction of of DNA damage long after UV exposurere delayed skin damage (light emission)n)

dark CPDs

We make beauty natural. Phytochemical Analysis

Objective: • To identify the main active ingredients of HerbaProtect NOX and link them to efficacy. Phenolic compounds, including flavonoids and tannins, are the main group of antioxidant phytochemicals due to their free radical scavenging activi- ties. Besides the direct scavenging activity, these phytochemicals also reduce oxidative stress by influencing enzymes involved in ROS and NO production [17, 18, 23-25] .

Technique: • Dereplication analysis of pomegranate flower extract: Liquid chromatography coupled with mass spectrometry as well as UV- and light scattering detectors (LC-MS/UV/ELSD) allowed database-assisted identification of secondary metabolites. • HPTLC (High performance thin layer chromatography) was used to assess the overall flavonoid/ polyphenol composition of HerbaProtect NOX. • Antioxidant power (AP), i.e. antioxidant capacity and reactivity were analyzed by electron spin resonance (ESR) spectros- copy. Unlike most other test systems, the AP method takes into account reduction potential and also reaction time (the lower the reaction time, the higher the activity).The antioxidant power is expressed as antioxidant units, where one unit corresponds to the activity of a 1 ppm solution of pure ascorbic acid as a benchmark [27] .

Granatin B 2.0 Punicacortein A / B Ellagic acid -5

1.5 Gallic acid Lagerstannin a Gemin B

1.0

c c d a b e f

Intensity [mAU]0.5 x10

1 2 3 4 5 6 7 8 Time [min] Granatin B Fig. 3: HerbaProtect NOX combines a wealth of potent metabolites. Dereplication analysis of pomegranate flower extract (left). The main components were identified as granatin B (17 %; right), ellagic acid (16 %), punicacortein A or B (12 %), lagerstannin A (9 %), gemin B (8 %), and gallic acid (4 %). Minor components denoted with (lowercase letters) were identified as hexahydroxydiphenoyl ester tannins such as gernaniin, rhoipteleanin B, or phyllanemblinin B.

Luteolin Apigenin Rosmarinic acid Gallic acid Antioxidative Power

AU Reaction Time

HerbaProtect NOX 7872 0.50

Market Reference * 148 1.00

Fig. 4: HerbaProtect NOX has a high-value polyphenol Fig. 5: HerbaProtect NOX has excellent antioxidative power. The antioxidative ca- and flavonoid profile. HPTLC fingerprint reveals pacity and reactivity of HerbaProtect NOX is tremendously superior compared to potent anitoxidants and free radical scavengers a market reference ( *an antioxidant complex designed to protect against oxidative such as rosmarinic acid or luteolin. stress, based on tocopherol and condensed tannins).

Results: • Pomegranate flowers are known to contain polyphenols, indicating potent antioxidant ability. Indeed, dereplication analy- sis of pomegranate flower extract revealed large amounts of polyphenols, mainly tannins (Fig. 3). • HPTLC fingerprint analysis of HerbaProtect NOX confirmed substantial amounts of phenolic compounds, e.g. rosmarinic and gallic acid, and flavonoids, e.g. luteolin and apigenin (Fig. 4). • The high proportion of free radical scavenging metabolites was reflected by an excellent antioxidative power, which was about 50-times higher compared to a reference product from the market (an antioxidant complex designed to protect against oxidative stress, based on tocopherol and condensed tannins; Fig. 5).

We make beauty natural. in vitro Activity

HerbaProtect ® NOX reduces peroxynitrite-mediated cell stress Objective: • To assess the peroxynitrite scavenging and antioxidant activity of HerbaProtect NOX. Peroxynitrite and ROS are key components in the formation of delayed sun-induced skin damage.

Technique: • DCF Assay: The non-fluorescing preliminary state of a dye diffuses into the cell and gets oxidized through ROS, resulting in a fluorescing product. An enhanced fluorescence signal thus indicates enhanced oxidative stress.

Study Details:

Design Cell-culture assay

Test Panel Primary human keratinocytes

Test Substances 0.006 % to 0.02 % HerbaProtect NOX

Application Frequency Pre-incubation with HerbaProtect NOX for 24 h followed by incubation with 40 mM peroxynitrite

(ONOOˉ) or 0.35 mM H 2O2 for 90 min.

Primary Endpoints Normalized fluorescence as a marker for oxidative stress

(A)  125

100 

75   

50   -induced oxidative stress [%] - ! # "   25 

 ONOO

0  untreated 0.006% 0.02% !    
 
HerbaProtect NOX HerbaProtect NOX  
  


Fig. 6: HerbaProtect NOX counteracts peroxynitrite- and H 2O 2 -induced skin stress. Human keratinocytes were stressed with peroxynitrite (A) or

H2O2 (B). HerbaProtect NOX markedly reduced the H 2O2-mediated oxidative stress level by 60% at only 0.02% and, more importantly, also showed specific protection against cellular stress mediated by peroxynitrite, a key driver in ‘dark CPD’ formation. N = 3; Mean ± SD; Student’s unpaired t-test versus untreated; * = p <0.05; ** = p <0.01.

Results: • HerbaProtect NOX showed specific protection against peroxynitrite-mediated cell stress and reduced oxidative stress in general (Fig. 6).

Conclusions: • HerbaProtect NOX reduces the negative effect of peroxynitrite and ROS exposure. This leads to a prevention of cellular damage in general, and more specifically, to reduced melanin excitation and subsequent ‘dark CPD’ formation.

We make beauty natural. in vitro Activity

HerbaProtect ® NOX counteracts the formation of ‘dark CPDs’

Objective: • To show that HerbaProtect NOX prevents the formation of DNA damage in sun-exposed skin. Cyclobutane pyrimidine di- mers (CPDs) are harmful DNA alterations that are typically created immediately upon UV-B absorption. However, around half of the CPDs occur several hours afterwards. Delayed CPDs can develop in the dark when peroxynitrite excites melanin. Excited melanin eventually induces ‘dark CPD’ formation by slowly transferring its excess energy to DNA [1] .

Technique: • CPD immunohistochemistry and signal quantification: Human skin biopsies were exposed to UV-A radiation and CPDs were quantified after 3 h. A decrease in CPDs could either be explained by a reduced generation of delayed ‘dark CPDs’, or by an improved repair of classical CPDs. The latter are built through direct UV absorption within picoseconds and are repaired subsequently over the next few hours.

Study Details:

Design Ex vivo study

Test Panel Abdominal skin biopsies from a 30-year old Caucasian female (3 skin discs/ condition with 3 quantifications each)

Test Substances 0.1 % HerbaProtect NOX

Application Frequency Curative setting to study repair effects: Topical application of HerbaProtect NOX immediately after UV-A irradiation (50 J/cm 2). CPD quantification 3 h after UV exposure

Primary Endpoints CPD labelling and quantification in the epidermis

* 1500

-22%

1000

UV-A

500 CPD abundance [%]

0 unstressed UV-A UV-A + UV-A + 0.1% HerbaProtect NOX 0.1% HerbaProtect NOX

Fig. 7: HerbaProtect NOX minimizes delayed skin damage, such as the Fig. 8: HerbaProtect NOX decreases DNA damage upon UV exposure. formation of ‘dark CPDs’. Representative skin sections with spe- CPD quantification by image analysis from ex vivo human skin biop- cific CPD labelling (brown area). CPD formation in the epidermis was sies. UV-A stress resulted in a ~13-fold CPD increase. HerbaProtect calculated by quantifying labelling intensity and area using software NOX application after UV-A stress reduced this increase by 22 %. analysis (see Fig. 8). N = 3; Mean ± SD; Student’s unpaired t-test; * = p <0.05.

Results: • A dose of 50 J/cm 2 of UV-A was used, as this dose induced an almost 13-fold increase in CPD formation, while lacking obvious cytotoxicity (verified by H/E staining; not shown). • Application of 0.1 % HerbaProtect NOX after UV-A stress significantly reduced CPD formation by 22 % (Fig. 7, 8).

Conclusions: • HerbaProtect NOX diminishes (delayed) skin damage upon sun exposure and offers a promising starting point for innovative cosmetic concepts that aim at reducing delayed skin stress and the associated premature skin aging.

We make beauty natural. in vivo Activity

HerbaProtect ® NOX provides immediate and long-term protection against sun-induced oxidative stress

Objective: • To evaluate the overall antioxidative protection of HerbaProtect NOX upon UV exposure. • To substantiate the mechanistic model of delayed sun-induced skin damage with peroxynitrite and excited melanin as active contributors.

Technique: • Quantitative determination of UV-A-induced chemiluminescence of human skin (ICL-S): The method evaluates oxida- tive stress within the skin. Free radicals damage cellular components and cause ultra-weak photon emission, which is detectable as a light signal. A decrease of the ICL signal thus indicates antioxidative potential [28] . • The method substantiates the mechanistic model of delayed sun-induced skin damage: Excited melanin is a key compo- nent in delayed skin damage, as it discharges its energy to DNA, e.g. causing ‘dark CPDs’. Alternatively, excited melanin discharges its energy by emitting light. A decrease of the ICL signal could thus also indicate a reduced melanin excitation.

Study Details:

Design Randomized, comparative, single-blind in vivo study

Test Panel 10 volunteers (8 female, 2 male); 42 – 65 years

Application Site Back

Test Substances Emulsion with 1 % HerbaProtect NOX (Sunset Lotion – request frame formulation for details)

Application Frequency Skin was stressed with 1 minimal erythemal dose (MED) of UV-A light followed by an immediate, single treatment with HerbaProtect NOX

Primary Endpoints ICL signal, i.e. ultra-weak light emission by the skin. ICL was measured in an acclimatized light- tight darkroom before, as well as 15 and 120 min. after treatment





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Fig. 9: HerbaProtect NOX increases the antioxidant capacity of the skin. Skin was stressed with UV-A followed by the application of 1 % HerbaProtect NOX (or untreated). Skin chemiluminescence, an indicator of oxidative stress and a marker for melanin excitation, was quantified 15 and 120 min. after treatment. UV-induced stress lasted for hours on untreated skin, whereas it was immediately and steadily reduced on skin treated with HerbaProtect NOX. N = 10; Mean ± SD; Student’s paired t-test compared to t=0 min.; * = p <0.05.

We make beauty natural. in vivo Activity

Results: • The application of HerbaProtect NOX resulted in an immediate and long-term reduction of skin chemiluminescence. 15 min. after UV-A irradiation, the ICL signal was reduced by 45 % compared to t=0 min. The effect remained stable for at least 2 h (Fig. 9).

Conclusions: • 1 % HerbaProtect NOX increases the skin’s antioxidant capacity for several hours and is therefore a valuable cosmetic active ingredient to:

° minimize free radical-related reactions ° reduce delayed sun-induced skin damage ° add an anti-aging effect to after-sun formulations ° support sunscreens or day creams with an extra line of defense ° protect the skin from infrared-A- or light-mediated premature aging

Product Characteristics

• HerbaProtect NOX is a unique concentrate of three plant extracts in a glycerol-based, preservative-free and self-pre- serving solvent system. This combination could be realized through a novel production method, which allows to combine and to concentrate hydrophilic and hydrophobic fractions of the extracts in a standardized, stable and easy-to-handle product. • Orange to brown liquid • Preservative-free/ self-preserving • COSMOS-approved raw material

Recommended Applications & Use Levels

• After-sun care, night repair, regenerating night masks • Day care, light protection, sun care • Antioxidant and radical scavenging products • Anti-aging formulations

Final cosmetic products with HerbaProtect NOX can be claimed as e.g. ‘Contains Swiss-manufactured ingredient‘. Recommended use level: 0.5 - 3 %

Safety

Toxicology: • Non-phototoxic (OECD 432) • Non-irritating for skin (HRIPT, repeated human patch test), when tested at a concentration of 10 % on 50 volunteers • Non-sensitizing for skin (HRIPT, repeated human patch test), when tested at a concentration of 10 % on 50 volunteers • Non-mutagenic (Ames test – OECD 471) • No allergens (as per current EU Cosmetic Regulation) • Moderately irritating for eyes (HET CAM), when tested at a concentration of 2.5 %. Considerations should be given to include ‘Avoid eye contact’ or equivalent in the instruction for use of the final cosmetic product, if applicable.

INCI

US: Glycerin, Water, Punica Granatum Flower Extract, Perilla Frutescens Leaf Extract, Terminalia Ferdinandiana Fruit Extract EU: Glycerin, Aqua, Punica Granatum Flower Extract, Perilla Frutescens Leaf Extract, Terminalia Ferdinandiana Fruit Extract For full and up-to-date INCI listing please see proprietary composition declaration (request under [email protected]).

We make beauty natural. Regulatory

Origin Punica Granatum Flower Extract: India * Perilla Frutescens Leaf Extract: China * Terminalia Ferdinandiana Fruit Extract: Australia * EU Cosmetic Regulation The product complies to the EU Cosmetic Regulation (EC) No. 1223/2009. China INCI All INCIs are listed in the current Inventory of Existing Cosmetic Ingredient China (IECIC). EU Reach The product, i.e. its substances conform to the Regulation (EC) No. 1907/2006. China Reach All ingredients conform to the legislation of China REACH. CMR The product does not contain substances classified as CMR under the Regulation (EC) No. 1272/2008 (CLP). ABS The plant materials used fully comply with the requirements of Access and Benefit Shar- ing (ABS) as derived from the Nagoya Protocol and its corresponding national legislation. Detailed information about the compliance is available ([email protected]). COSMOS HerbaProtect NOX is a raw material approved by ECOCERT Greenlife that conforms to the COSMOS Standard. The raw material is of 100 % natural origin.

Natural/organic HerbaProtect NOX can be used in natural, organic and natural/ organic certified cosme- tics. Halal All ingredients conform to HALAL requirement, considering the following: Traces of ethanol remaining in HerbaProtect NOX at the end of the manufacturing process are technically unavoidable. The ethanol is plant-derived and non-GMO. Its concentration is equal or less than 0.001 % at a maximal recommended use level of max. 3 % HerbaProtect NOX in the final cosmetic product. Vegan The product can be used in vegan formulations. Non-GMO The product is non-GMO. It meets the non-GMO standards set by Regulation (EC) No. 1829/2003. Palm oil The product does generally not contain palm (kernel) oil or its derivatives. * Not part of the specification References

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Kochevar, Nox1-based NADPH oxidase is the major source of UVA-induced reactive oxygen species in human keratinocytes. J Invest Dermatol 128, 214-222 (2008). 27. K. Jung et al., The antioxidative power AP--A new quantitative time dependent (2D) parameter for the determination of the antioxidant capacity and reactivity of different plants. Spectrochim Acta A Mol Biomol Spectrosc 63, 846-850 (2006). 28. M. Rohr et al., Influence of repetitive UVA stimulation on skin protection capacity and antioxidant efficacy. Skin Pharmacol Physiol 24, 300-304 (2011). Disclaimer: Please be aware that the listed properties (medicinal or otherwise) that have been sourced from literature should be understood as value-added information only. No proof of statements by testing or otherwise will be provided by Lipoid Kosmetik AG (this excludes test data generated by Lipoid Kosmetik AG in support of our specific actives range). Other product properties identified and highlighted by specific tests or studies are to be interpreted in the context of the test/study conditions only. Product properties mentioned herein going beyond the product specifications are applicable unless the inherent uncertainty regarding the availability of herbal raw material (or its derivatives) compels a modification. Please be aware that the use of any claim on cosmetic products is the sole responsibility of the customer and is regulated by your local Regulatory Body.

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