Dermatological application of soy isoflavones to prevent skin aging in postmenopausal women

Dermatological Application
of Soy Isoflavones to
Prevent Skin Ageing in
Postmenopausal Women
Schmid Daniel, Muggli Reto and Zülli Fred Mibelle AG Cosmetics, Bolimattstrasse 1, 5033 Buchs, Switzerland Abstract
genistin and daidzin (Fig. 1 and 2). In nonfermented
Isoflavones from soy beans are used in functional soyfoods, they occur predominantly in form of the food products as a measure to alleviate hormone- polar, water-soluble glycosides, whereas fermented soy products contain mainly the biologically active phytohormones are well perceived and do not showthe negative side effects found in conventionaltherapies with hormones such as estrogens. Anumber of studies show that estrogens can slow downthe aging process of the skin in postmenopausalwomen. For this reason, isoflavones from soy beansare also very interesting compounds for cosmeticformulations. We succeeded in isolating very purefractions of isoflavones in the form of the activeaglycones suitable for cosmetic applications. Weherein report the use of this aglycone preparation inthe treatment of post menopause skin and discussfurther applications.
Phytoestrogens are plant derived compounds with
Fig. 1. Molecular structures of the hormone estrogen and biological activities comparable to the human
the phytoestrogens genistein and daidzein hormone estrogen
Epidemiological studies indicating associations between diet and disease states led to theinvestigation of a series of bioactive plant compounds,called phytochemicals. Some of these nonnutrientplant chemicals, such as phytoestrogens were shownto confer significant long-term health benefits.
Compound
Phytoestrogens are structurally and functionally similar to human estrogen (Fig. 1). There are two
types of phytoestrogens, the lignans and the isoflavones. Soybeans are especially rich inisoflavones. They contain the isoflavones genistein Fig. 2. Molecular structures of the isoflavone glycosides of and daidzein and their respective β- g l y c o s i d e s , The role of the hormone estrogen and the
mechanism of its action
cardiovascular system. Thus the compound should Estrogen influences growth, differentiation and act as a non- or anti-estrogen in breast and uterus function of the female and male reproductive system (Table 1). Recently a second estrogen receptor (ERβ)
such as mammary gland, uterus, vagina, ovary, testes, was found in addition to the classical (ERα) one. The epididymis, and prostate as well as other tissues tissue distribution and the ligand binding affinities of (Korach et al. 1995). Estrogen plays an important role the two receptors are different (Kuiper et al. 1997 in the development and maintenance of the bone The effectiveness of selective estrogen receptor structure as well as for the cardiovascular system modulators depends on their uptake by the various (Turner et al. 1994, Farhat et al. 1996 tissues and on their specific affinities to the two principal forms of estrogens, which are produced mainly in the ovaries and testis, are 17β-estradiol, themost active form, estrone and estriol. Upon arrival at Phytoestrogens as natural alternative to estrogen
the target cell the estrogens are retained by replacement therapies
intracellular binding proteins, the estrogen receptors Isoflavones are currently intensively studied as (ER). The saturated ER undergoes a conformational natural alternatives to human estrogens used in change that allows the receptor to interact with the replacement therapies. The fact that Asian women, DNA and to modulate the transcription of target that traditionally consume a lot of soy products, are less affected by the menopausal symptoms, such asosteoporosis, cardiovascular disease, and breast Estrogen-related physiological disorders and their
cancer, is attributed to their high isoflavone intake As the ovaries age, their function declines until with soy isoflavones - so called ‘functional foods’ menopause where the production of estrogen and or nutraceuticals - are claimed to prevent progesterone drops drastically. These hormonal postmenopausal problems, especially osteoporosis.
changes lead to menopausal symptoms like hot The structures of isoflavones, heterocyclic phenols, flashes which have a severe negative impact on many are very similar to the steroidal estrogen (see Fig. 1).
women's quality of life. In addition, they are Thus they can bind to some extent to estrogen considered to be a potential risk factor for osteoporosis and cardiovascular diseases (Utian 1989).
Compared with the principal circulating estrogens Hormone replacement therapies have been found to in humans, isoflavones are bound at a much lower help against these menopause-related disorders.
rate (103-fold weaker), however, they do compete with However, the continuous administration of estrogen estrogens for receptor sites. Recent data show that is sometimes associated with severe adverse effects phytoestrogens have higher affinities to ERβ than to such as vaginal bleeding and in rare cases with the ERα. Phytoestrogens have been shown to protect development of breast and uterus cancer (Breckwoldt against osteoporosis and heart disease without1 9 9 5). Synthetic estrogen-like compounds, called affecting the uterus and breast (Kuiper et al. 1998, selective estrogen receptor modulators (e.g.
Cassidy 1999). Clinically, isoflavones may behave like r a l o x i f e n e®), were introduced recently for an selective estrogen receptor modulators. Thus, the improved hormone replacement therapy (Compston application of isoflavones may have decisive1 9 9 8). The perfect drug should have a tissue advantages over classical hormone replacement selectivity and exert an estrogenic effect only in Table 1. Selective tissue effects of synthetic estrogen-like compounds (e.g. raloxifene) and isoflavones (Maroulis 2000). Benefits of phytoestrogens in skin care
oil. Thus, a special galenic form is necessary to After menopause, the skin undergoes changes that introduce these isoflavone preparations into cosmetic include thinning, decrease in elasticity and wrinkle aging is the consequence of a lower production of isoflavones suitable for cosmetic applications (Fig. 3).
collagen and elastin, the supportive and elasticproteins of the skin (Brincat et al. 1987, Affinito et al.
1999, Adamiak et al. 2000 be an organ responding to ovarian hormones. Indeedestrogen receptors have been identified in humanskin (Hasselquist et al. 1980, Schmidt et al. 1990 question as to how far this skin aging is induced bypostmenopausal hypoestrogenism is still underdiscussion (Affinito et al. 1999). Several studies show that estrogen therapy preserves collagen content, elastic properties, andthickness of the skin in postmenopausal women(Hasselquist et al. 1980, Punnonen et al. 1987, Castelo-Branco et al. 1992, Dunn et al. 1997 Fig. 3. Production process for a cosmetic phytoestrogen treatments were shown to reverse postmenopausal skin aging (Callens et al. 1996, Schmidt et al. 1996 Estrogenic compounds reduce the depth of wrinkles A soy preparation which is rich in isoflavones is and increase the tone and hydration of the skin. treated with a βglucosidase to deglycosylate the Since the results obtained with isoflavone isolflavones to the corresponding aglycones. As a supplemented nutraceuticals are very positive, we consequence of their hydrophobic nature, these believe that the topical application of phytohormones aglycones precipitate almost entirely and can be can also fight some of the menopause-associated skin separated by centrifugation from the enzyme and the problems. In in vitro testing genistein stimulated water-soluble brown polyphenols. The aglycones are collagen synthesis (Kawashima et al. 1996 extracted with alcohol to get a phytohormone regulated matrix metalloproteinases (Shao et al. ) .
solution. The aglycones can then be incorporated into Therefore topical application of isoflavones is not liposomes to get a water dispersable preparation only beneficial in postmenopausal skin, where it which has a high bioavailability in the skin.
replenishes the loss of endogenous estrogen but alsoin normal skin when used to treat cellulite or dull skin Analytical characterization of isoflavones
Isoflavones have an absorption maximum at 260 nm and can be separated on a reverse phase column.
Development of a pure isoflavone preparation
Therefore the aglycone formation in our purification suitable for skin care
process can be followed by HPLC analysis. The There are several soy isoflavone products on the analysis of the starting material revealed that the market that are sold as dietary food supplements. In isoflavone glycosides daidzin and genistin are the most cases, these food supplements contain only main components. After the β-glucosidase treatment, isoflavone glycosides, the molecular form that is these glycosides were completely transformed to their biologically not active. However, after ingestion the corresponding aglycones daidzein and genistein (Fig.
glycosides are transformed by intestinal glucosidases 4). The Genistin of the source material could be
and intestinal bacterial metabolism into the recovered at 86% as genistein and daidzin at 50% as estrogenically active form (Setchell and Cassidy 1999 daidzein respectively after alcohol extraction. The Since the skin does not habour such bacteria and characterization of the aglycone solution in alcohol enzymes, the active isoflavone preparations for skin revealed that the product has a purity of > 80%.
care must be in the form of aclycones. Unfortunately, Upon dilution of this alcohol solution in water white these aglycones have a poor solubility in water and needles of geinstein and daizein became visible.
lotions and creams since the product can easily beformulated into the water phase of cosmetics. Acombination with moisturizers and classical anti-agingingredients, such as vitamins and antioxidants mayeven further improve the activity of the product. It is evident, that isoflavones are also very interesting compounds for anti-cellulite products asthey enhance the skin tone and its viscoelasticproperties. Thus, we use our aglycone preparation inthe respective field. In our products, we combinedthe phytohormones with an algae extract (spirulinaplatensis) which is rich in minerals to achieve a highly Fig. 4. HPLC analysis of isoflavones before (blue) and efficient anti-cellulite formulation suitable to treat -glucosidase treatment. Separation was done mone dependent skin and subcutis problems on a Merck Superspher 100 RP-18C column (4 µm, 254 Zülli et al. 2001).
x 4 mm) at 40°C. Solvant A was H2O + 0.1% acetic acid and solvant B acetonitril + 0.1% acetic acid. A gradientisoflavones have been reported (Setchell and Cassidyfrom 10% B to 30% B in 60 min was applied. Flow was 1999). But only little data exist about the effects of soy 1.5 ml/min and detection was at 260 nm. isoflavones on histologically normal human breast. Inone of these very intersting studies McMichael- Application of soy isoflavone aglycones in
Phillips et al. 1998 found that phytoestrogens can cosmetic formulations
stimulate the proliferation of breast epithelium Over the last decade, plant derived ingredients significantly in vivo based on higher serum levels of have gained a lot of interest in cosmetic formulations.
genistein and daidzein after soy supplementation.
However, in many cases, these products do not offer Thus the application of liposomal aglycones in breast the proposed activity since the preparations do not firming products is a new opportunity to improve contain the active molecules in an appropriate skin tone and elasticity in women of all ages.
concentration or the compounds are not bioavailable.
The described liposomal aglycone preparation is anew active ingredient derived from soy. The References
preparation contains the active form of isoflavones in Adamiak, A., Skorupski, P., Rechberg e r, T. & a high concentration and offers a number of different Jakowicki, J. A. (2000) The expression of the gene cosmetic and dermatological applications. encoding pro-α 1 chain of type I collagen in the skin Oral and topical estrogen applications have shown of premenopausal and postmenopausal women. Eur.
to preserve collagen content, elastic properties, and J. Obstet. Gynecol. Reprod. Biol. 93: 9-11.
thickness of the skin in postmenopausal women Adlercreutz, H., Hamalainen, E., Gorbach, S. & (Dunn et al. 1997, Schmidt et al. 1996 Goldin, B. (1992) Dietary phyto-oestrogens and the human hormones cannot be used in cosmetic menopause in Japan. Lancet 339: 1233.
formulations and due to the potential adverse effects Affinito, P., Palomba, S., Sorrentino, C., Di Carlo, should only be considered in severe cases in C., Bifulco, G., Arienzo, M. P. & Nappi, C. (1999) dermatological products. The treatment of skin Effects of postmenopausal hypoestrogenism on skin during and after menopause with an active aglycone preparation from soy is a completly new approach in Bolognia, J. L., Braverman, I. M., Rousseau, M. E.
the battle against the aging process in the skin. The & Sarrel, P. M. (1989) Skin changes in menopause.
loss of hormonal activity reduces the skin tone and its hydration. This deterioration is based on the Breckwoldt, M., Keck, C. & Karck, U. (1995) reduction of the metabolic activity of skin cells and Benefits and risks of hormone replacement therapy.
J. Steroid Biochem. Mol. Biol. 53: 205-208. Isoflavones aglycones encapsulated into liposomes Brincat, M., Kabalan, S., Studd, J. W. W., Moniz, C.
can be applied in different formulations such as gels, F., de Trafford, J, & Montgomery, J. (1987) A Study of the Decrease of Skin Collagen Content, Skin Estrogen Receptor α and β. Endocrinology 138: 863- Thickness, and Bone Mass in the Postmenopausal Woman. Obstet. Gynecol. 70: 840-845.
Kuiper, G. G. J. M., Lemmen, J. G., Carlsson, B., Callens, A., Vaillant, L., Lecomte, P., Berson, M., Corton, J. C., Safe, S. H., Van Der Saag, P. T., Van Der Gall, Y. & Lorette, G. (1996) Does Hormonal Skin Burg, B. & Gustafsson, J.-A. (1998) Interaction of Aging Exist? A Study of the Influence of Different Estrogenic Chemicals and Phytoestrogens with Estrogen Receptor β. Endocrinology 139: 4252-4263.
Maroulis, G. B. (2000) Alternatives to Estrogen Measurement Techniques. Dermatology 193: 289- Replacement Therapy. Ann. N.Y. Acad. Sci. 900: 413- Cassidy, A. (1999) Potential tissue selectivity of McMichael-Phillips, D. F., Harding, C., Morton, dietary phytoestrogens and estrogens. Curr. Opin.
M., Roberts, S. A., Howell, A., Potten, C. S. & Bundred, N. J. (1998) Effects of soy-protein Castelo-Branco, C., Duran, M. & Gonzalez-Merlo, supplementation on epithelial proliferation in the J. (1992) Skin collagen changes related to age and histologically normal human breast. Am. J. Clin.
hormone replacement therapy. Maturitas, 15: 113- Punnonen, R., Vaajalahti, P. & Teisala, K. (1987) Compston, J. E. (1998) Selective oestrogen Local Oestriol Treatment Improves the Structure of Elastic Fibers in the Skin of Postmenopausal Women.
applications. Clin. Endocrinol. 48: 389-391.
Dunn, L. B., Damesyn, M., Moore, A. A., Reuben, Schmidt, J. B., Lindmaier, A. & Spona, J. (1990) D. B. & Greendale, G. A. (1997) Does Estrogen Hormone Receptors in Pubic Skin of Premenopausal Prevent Skin Aging? Arch. Dermatol. 133: 339-342.
and Postmenopausal Females. Gynecol. Obstet.
Farhat, M. Y., Lavigne, M. C. & Ramwell, P. W.
(1996) The vascular protective effects of estrogen.
Schmidt, J. B., Binder, M., Demschik, G., Bieglmayer, C. & Reiner, A. (1996) Treatment of Skin Hasselquist, M. B., Goldberg, N., Schroeter, A. & Aging with Topical Estrogens. Int. J. Dermatol. 35: Spelsberg, T. C. (1980) Isolation and Characterization of the Estrogen Receptor in Human Skin. J. Clin.
Setchell, K. D. R. & Cassidy, A. (1999) Dietary Isoflavones: Biological Effects and Relevance to Jensen, E. V. (1995) Steroid hormones, receptors Human Health. J. Nutr. 129: 758S-767S.
and antagonists. Ann. NY Acad. Sci. 761: 1-17. Shao, Z. M., Wu, J., Shen, Z. Z. & Barsky, S. H.
Kawashima, K., Inoue, T., Tsutsumi, N. & Endo, (1998) Genistein inhibits both constitutive and EGF- H. (1996) Effect of KCA-098 on the Function of stimulated invasion in ER-negative human breast Osteoblast-Like Cells and the Formation of TRAP- carcinoma cell lines. Anticancer Res. 18: 1435-1439.
Positive Multinucleated Cells in a Mouse Bone Turner, R. T., Riggs, B. L. & Spelsberg, T. C.
Marrow Cell Population. Biochem. Pharmacol. 51: (1994) Skeletal effects of estrogens. Endocr. Rev. 15: Korach, K. S., Migliaccio, S. & Davis, V. L. (1995) Utian, W. H. (1989) Biosynthesis and physiologic Estrogens. In: Munson P. L. (ed) Principles of effects of estrogen and pathophysiologic effects of P h a r m a c o l o g y. Basic Concepts and Clinical estrogen deficiency: a review. Am. J. Obstet. Gynecol.
Applications. Chapman and Hall, New York, pp 809- Wang, H. & Murphy, P. A. (1994) Isoflavone Kuiper, G. G. J. M., Carlsson, B., Grandien, K., Content in Commercial Soybean Foods. J. Agric.
Enmark, E., Häggblad, J., Nilsson, S. & Gustafsson, J.-A. (1997) Comparison of the Ligand Binding Zülli, F., Muggli, R. & Hanay, C. (2001) Patent Specificity and Transcript Tissue Distribution of

Source: http://www.mibellebiochemistry.ch/pdfs/Dermatological_Application_of_Soy_Isoflavones_to_prevent_skin_aging_in_postmenopausal_women_CTMWW_20Dermatological_A.pdf