Natural ingredient complex evens and lightens skin

 It is believed that whiter, more evenly-toned skin can hide physically unattractive facial features create a perception of younger, more beautiful overall appearance. This is why much effort is invested by the cosmetic industry to make skin whitening easier and the products more effective and safe. Thus an alternative, which would be safe, while its scientifically backed activity is well-established, is ever more desired though challenging to find. Towards this goal, IBR presents a novel complex of ingredients – IBR Clearly Bright with effective skin lightening effect. IBR Clearly Bright is a combination of patented actives by IBR Ltd – IBR-Phyto(flu)ene, IBR-Dormin and IBR-Snowflake. Recent clinical findings supports the mechanism of action previously established in vitro of IBR Clearly Bright Complex as an outstanding and effective solution for the challenge of skin whitening, lightening and evening of skin tone.

 Hyperpigmentation is the darkening of skin areas, caused by increased melanocyte proliferation, melanin synthesis and/or melanin deposition (Fig. 1a). Although known to be associated with numerous physiological conditions, hyperpigmentation is most closely linked to repeated irritation and inflammation of the skin. This inflammation is most commonly initiated by exposure to UV. Skin colour is primarily determined by a number of closely related factors: melanocyte distribution in the skin, the content level of melanin within these melanocytes and the transfer and distribution of melanin to the keratinocytes surrounding melanocytes. Ageing, acne, depilation, sun exposure, hormonal imbalance, various skin disorders form just a short list of repeated insults to the skin that result in hyperpigmentation. Such hyperpigmentation is presented as darker skin tone, dark spots (age spots), freckles and uneven skin tone. The frequency of these undesired phenomena has driven science to seek a solution. Preventive methods such as sun exposure avoidance and sunscreen use, attempt to reduce the UV contribution to this undesired effect. Whitening or bleaching agents are implemented for the elimination of dark spots and lightening of skin tone. These agents act on the various steps involved in the pigmentation pathway. To aid in this mission IBR presents a novel complex of ingredients. IBR Clearly Bright is a combination of patented technologies by IBR Ltd – IBR-Phyto(flu)ene, IBR-Dormin and IBR-Snowflake. This complex addresses the skin darkening process in its various steps (Fig 1b). Recent clinical findings present it as an effective solution for the challenge of skin whitening, lightening and evening of skin tone.

The actives: concepts in brief

The colourless carotenoids phytoene and phytofluene

Phytoene and phytofluene, branded IBR-Phyto(flu)ene by IBR, are the first carotenoid precursors in the biosynthetic pathway, progenitors of all other carotenoids. These molecules are devoid of visible colour, absorbing light in the UV range only, thus appearing colourless to the human eye. These colourless carotenoids are found in most carotenogenic organisms including various microorganisms, algae, fungi and plants, where they act as an intrinsic shield against excessive radiation and other environmental stress. Phytoene and phytofluene have been demonstrated to accumulate in skin, preferentially as a result of a carotenoidsrich diet. This allows the skin similar protection to the one granted by the colourless carotenoids to plants and other organisms. Phytoene and phytofluene together bring into the complex the beneficial efficacy of enhanced skin anti inflammatory capacity and protection from UV and oxidative stress. Recently, the ingredient’s isolated effect on skin lightening was demonstrated by a clinical study. Its technology comprises natural extracts from microalgae (IBR-CLC) or tomato (IBR-TCLC) sources for topical application. Phytofloral, a nutriceutical, provides a solution for beauty from within. All three platforms, IBR-CLC, IBR-TCLC and Phytofloral, are rich in the colourless carotenoids phytoene and phytofluene. Activity of these three members in relation to control of pigmentation has been established as (a) inhibition of melanin synthesis (Table 1), (b) absorbance of UV light both in the UVB and UVA (Fig. 3), (c) reduction in transmittance of the UVB and UVA, (d) anti inflammatory activity (e) excellent anti oxidation mostly against hydroxyl radical (Fig. 2) and (f) prevention of damage to DNA (Table 2).

Narcissus tazetta bulb extract

IBR-Dormin’s ability to slow down melanocytes proliferation derived from its Dormins activity was established. Further, its inhibitory effect on endothelin 1 results in inhibition of melanin synthesis and melanocytes proliferation (Fig. 1b). Dormins are natural extracts from plants and plant organs in their dormant stage which are able to slow down cell proliferation to maintain younger, healthier skin. This ingredient captures and transfers flower bulbs dormancy and rejuvenation to skin. Flower bulbs become dormant through the winter into what may be pictured as a “beauty sleep” to allow beautiful, rejuvenated blossom in the spring. IBR’s anti-ageing technology via implementation of Dormins features slow-down of skin ageing as a result of slowing down cell proliferation, thereby preserving youth cell capital and improving skin appearance and maturation.

Leucojum aestivum bulb extract

By its ability to boost Super Oxide Dismutase (SOD) expression in the skin, IBR-Snowflake enhances the skin’s ability to defend itself from oxidative damage that, in many cases, is so closely related to pigmentation (Fig. 5). Along with its ability to slowdown melanocytes proliferation and providing the skin its “beautifying sleep”, the ingredient contributes to the reduction in the amount of total melanin in the skin, resulting in lighter and more even skin tone (Fig. 1b). It is a natural, aqueous extract from dormant summer snowflake bulbs, and slows down skin ageing by reduction of cell proliferation through dormancy and induction of skin’s Super Oxide Dismutase (SOD) inherent defence mechanism. The extract was clinically shown to smoothen the skin and reduce wrinkles via inhibition of muscle cell contractions, contributing to brightening and whitening of the skin through dormancy of the melanocytes.

Activity of phytoene and phytofluene

Skin colour is primarily determined by the amount of the pigment melanin present in melanocytes. Melanin is produced in response to the stimulus of UVA light, which leads to the well-known tanning effect of the skin. Ageing, exposure to sun, hormonal abnormalities, and various skin disorders increase the deposition of the melanin pigment in the skin, resulting in dark spots and freckles. Such dark spots are not only considered to be unattractive in many societies, they are also known to be unhealthy and, therefore, undesired. For those seeking elimination of dark spots or lightening of skin tone, whitening or bleaching compositions are useful. Typical whitening agents in cosmetic formulations are kojic acid, arbutin, licorice extract, and vitamin C. These agents act as either effective tyrosinase inhibitors or anti-oxidants, but some are unstable, easily oxidised and degraded. Moreover, the risk of skin irritation and subsequent inflammation may result in an opposite effect to that which is desired. Phytoene and phytofluene work to lighten skin via the following methods:

• Inhibition of melanin synthesis, a direct activity (Table 1).
• Control of pigmentation by antiinflammatory activity.
• Reduction of melanin production by reduction of oxidative stress (Fig. 2, Table 2).
•  Reduction of skin darkening due to sun exposure by UVA/B absorption (Fig. 3).
•  Pigmentation control by reduction of UVB induced inflammation by UVB absorption.

Effect on melanin synthesis

The effect of IBR-TCLC (tomato phytoene and phytofluene) on melanin synthesis in cultures of normal human melanocytes and melanoma cells was tested. In one study, the compound IBR-TCLC tested at 0.5% decreased to 71% the melanin content of normal human melanocytes as compared to control In addition, for evaluation of the whitening effect of tomato phytoene and phytofluene in squalane 0702 on melanoma cells, said cells were treated with IBR-TCLC in squalane 0702 (0.25%, 0.50%, 1.25%, 3.75%, 5.00%) for 96 hours and harvested. Harvested cells were centrifuged and the colour of the cell pellets was evaluated. Cells treated with over 0.5% tomato phytoene and phytofluene in squalane 0702 showed a whitening effect in a dose response increased manner (Table 1).

Anti-inflammatory activity

Reduction of inflammation on areas susceptible to repeated aggravation (like shaving or chafing for example) is suggested to reduce skin pigmentation. Anti-inflammatory activity of the IBR-CLC (colourless carotenoids from Alga Dunaliella sp.) was established. Significant reduction in expression of IL-6 and IL-12 in activated lymphocytes was demonstrated. No LDH leakage was seen indicating no loss of viability at the doses tested [Oppen- Bezalel L et al. Fragrance Journal 2006; 34 (3): 48-53]. A significant reduction in PGE-2 expression in IL-1 induced normal human fibroblasts of 46% by colourless carotenoids from Alga Dunaliella sp. was observed and published [Fuller BB et al. J Cos Derm 2006; 5 (1): 30-38].

Antioxidant activity

Colourless carotenoids quench hydroxyl radicals – some of the most destructive free radicals in nature. We propose use of the above-mentioned extracts for reduction of oxidative stress, linked to inflammatory conditions and damage to DNA that induce melanin synthesis leading to hyperpigmentation and skin damage.

DNA protection

In the study results (Table 2) phytoene and phytofluene provide an effective DNA protection activity against free radical damage.

Narcissus tazetta bulb extract activity

Narcissus tazetta bulb extract’s evident activity, natural source and lack of known side effects make it an ideal candidate for a skin whitening agent. Its methods of activity are as follows:

• Inhibition of melanocytes proliferation (Fig. 4).
• Control of melanin synthesis and pigmentation by inhibition of endothelin 1 (ET-1), whose expression in keratinocytes is induced by UV.
• Reduction of melanin production by reduction of oxidative stress affecting the nitric oxide pathway (Table 3).

Endothellin-1 production inhibitory effect

Narcissus tazetta bulb extract (IBR-Dormin) potently inhibits endothelin-1 production by normal human keratinocytes in vitro. Lasserre C, Mildner M, Wallaert C, Tschachler E. Skin Research and Technology 2004; 10: 1–16. Abstracts, 2004 US Symposium of the International Society for Bioengineering and the Skin staes: “Among several samples tested Narcissus tazetta bulb extract was found to significantly inhibit ET-1 production/release in a dose dependent manner. In addition this extract also completely inhibited the IL1a-induced up-regulation of ET1 production by normal human keratinocytes in culture. Therefore Narcissus tazetta bulb extract represents an interesting active ingredient for products used to attenuate and prevent pigment spots of the skin.”

Nitric oxide (NO) inhibitory effect

Griess Assay – background: Nitric oxide is a transducing molecule (and a gas) that has been demonstrated to be involved in physiological processes such as vasodilatation and neurotransmission as well as in pathological processes such as inflammation and cancer. Higher NO levels have been found in psoriasis. It is also well established that high NO concentrations are toxic for the tissues. NO combines with superoxide radical and forms peroxynitrite, a highly toxic free radical species. Applications for materials that decrease NO levels include the following: redness, inflamed skin, vasodilatation, prevention of environmental damage. Applications for materials that increase NO levels include the following: hair growth, pigmentation, improved wound healing. Narcissus tazetta bulb extract inhibits LPS-induced nitric oxide production in (LPS) activated murine macrophages with an IC50 of 5.6 ?g/mL. Compared to quercetin (a reference-inhibitor), Narcissus tazetta bulb extract is a more potent inhibitor.

Leucojum aestivum bulb extract activity

Leucojum aestivum bulb extract’s evident activity, natural source and lack of known side effects make it an ideal candidate for a skin whitening agent. Its methods are as follows:

•  Inhibition of melanocytes proliferation.
• Natural control of melanin synthesis and pigmentation – reducing oxidative damage by boosting Super Oxide Dismutase (SOD) expression leading to elimination of SO ions (Fig. 5).

SOD expression increase

In nature, skin eliminates oxidative stress through enzymes like SOD. Leocojum aestivum bulb extract increased the expression of MnSOD enzyme significantly in keratinocytes at a low concentration. The protein expression was evaluated by flow cytometry (fluorescence-activated cell sorting or FACS) on normal human epidermal keratinocytes (NHEK) cells in culture. This study was based on published tests methods. Leucojum aestivum bulb extract was found to increase the expression of the antioxidant enzyme MnSOD in keratinocytes using FACS-based testing system with fluorescently marked MnSOD antibodies. The extract was added to the cell based test system in several increased concentrations compared with culture media control which was used as the base line for the expression of the SOD. The effect of the extract was significant (p<0.05) at a concentration of 0.15% extract giving a 20% increase in the SOD expression. Leucojum aestivum bulb extract significantly increases MnSOD in keratinocytes (*20% increase at 0.15% Leucojum aestivum bulb extract) improving the natural skin protection and reducing oxidative damages.

Clearly Bright complex: clinical evaluation

A randomised, double blinded, placebocontrolled study evaluating the lightening effect of a cream containing tomato phytoene and phytofluene 5% and cream containing tomato phytoene and phytofluene 3%, Narcissus tazetta bulb extract 2% and Leucojum aestivum bulb extract 2% (Clearly Bright complex) was performed. A twice a day application during a three month course of treatment resulted in a significant skin whitening effect in both Day 42 and Day 84 (n=21) (Figs. 6-8).

Clinical study conclusions

The complex induced a significant lightening effect associated to a more improvement in the visual cutaneous state than placebo (Figs. 9 & 10).

Conclusion

The demonstrated report and studies substantiate Clearly Bright Complex, a complex comprising: IBR-Dormin all natural 0305, IBR-Snowflake all natural 1003 & IBRTCLC in jojoba oil 0705 [IBR-Phyto(flu)ene] as most effective and significantly able to lighten and whiten the skin tone. The complex acts on whitening and lightening the skin tone through multiple mechanisms and pathways, making it an ideal active complex addressing skin whitening, protection, health and beauty in a complete approach that is safe, active and natural.