MXPA00007390A - Reducing hair growth, hair follicle and hair shaft size and hair pigmentation - Google Patents

Abstract

A topically effective composition for effecting changes in mammalian hair growth, hair pigmentation and hair shaft and follicle size comprises one or more compounds derived from one or more of the botanical families leguminosae, solanaceae, gramineae and cucurbitaceae. The composition preferably contains one or more serine protease inhibitory compounds.

Description

REDUCTION OF HAIR GROWTH, FLEMIC SIZE AND PILES, AND PIGMENTATION OF THE HAIR FIELD OF THE INVENTION This invention relates to effective methods and compositions for reducing hair growth. Very specifically, the present invention relates to methods for modifying the speed of hair growth, reducing the size of the hair follicle and the hair shaft, as well as for reducing the pigmentation of the hair shaft, by topically applying both botanical extracts that they contain an inhibitory activity of serine protease and in particular soy extracts such as soy milk, or mixtures and formulations of the foregoing, combined with other active ingredients such as isoflavones.

BACKGROUND OF THE INVENTION A primary function of hair and hair of mammals is to provide protection against the environment. However, this function has been lost considerably in humans, in which the hair is maintained or removed essentially for social and cosmetic purposes.

Various procedures are used to remove unwanted hair, including shaving, electrolysis, hair removal, laser and light beam therapies, as well as the injection of therapeutic antiandrogens. These conventional methods have their drawbacks. For example, shaving can cause cuts and cracks on the surface of the skin, can leave a feeling of increased speed of new hair growth, and can also leave a feeling of undesirable stubble. Although electrolysis can maintain a certain area free of unwanted hair for a prolonged period, the procedure is often very expensive and painful and can also cause scarring. Not only can hair removal cause pain and discomfort, it also causes the very weak removal of short hair. Various unwanted side effects, such as effects on muscularity, frequently accompany the use of antiandrogens. For these reasons, better methods are required to reduce hair growth. Beard pseudofolliculitis (Pseudofolliculitis barbae) is an inflammatory hair disorder that is most commonly found in the area of the beard. It causes inflammation of the follicular papillae when the ends of the hair penetrate the skin instead of passing through the follicular orifice. This procedure is extremely common in black-skinned men because their hair is often curly, which comes out of the skin at an acute angle. When shaving very close to the skin, particularly with a razor, predisposes them to pseudo-folliculitis of the beard. The most effective treatment available is to let the hair grow beyond the surface of the skin. Often, such treatment is not desired. Hirsutism is a relatively common condition that affects approximately 4% of women. Facial hirsutism frequently interferes with personal and work activities, and temporary hair removal is a major component in the management of patients with hirsutism. Shaving is the temporary method that is used most frequently to eliminate facial hair, such as waxing, waxing and depilating agents that are more difficult to tolerate and must be very careful to avoid folliculitis, pigmentation and scars. . Cosmetic makeup is generally used to hide cuts and the stubble beard; In addition, electrolysis and thermolysis can be used for permanent hair removal when available. An alternative or complementary approach desired for hair removal would be a method to reduce hair growth, reduce the hair follicle, the size of the hair shaft, as well as the pigmentation of the hair shaft. This approach could reduce the visibility of existing hair, making it smoother and lighter. When combined with other methods of hair removal, this method could improve and prolong the effect of removal, and thus reduce the need and frequency of hair removal. The long-term use of such an approach could lead to the attenuated, mild growth of the hair with the reduction of pigmentation, which is less visible and therefore does not require the use of other removal methods. The reduction of hair growth in the underarm area (fossa axillaris) is desired, where deodorants and antiperspirants are used to control odor trapped inside the axillary hairs. It would be desirable to have products for use in the armpits, which combine deodorants with antiperspirants and which provide a hair reduction activity. The hair of African type is unique in its morphology, a curly hair shaft with variations in diameter. This complex stem structure gives rise to the need for specialized cleaning and maintenance products and procedures to ensure that African-style hair maintains the desired cosmetic properties. It is desired to have products that reduce this complexity and that make African hair more manageable, thus improving its appearance. It would also be desirable to provide a method for affecting the growth of hair chemically or naturally, the hair follicle, the size of the hair shaft, as well as the pigmentation of the hair shaft, which does not cause undesirable side effects for the user. .

BRIEF DESCRIPTION OF THE INVENTION According to the present invention, compositions and methods have been discovered which cause changes in the growth of hair and hair of a mammal, the hair follicle, the size of the hair shaft, as well as the pigmentation of the hair, by means of topical application to hair. the skin of a mammal, of an effective amount of a topically active composition consisting of protease inhibitors, botanical extracts and in particular legume extracts including, but not limited to, soy milk, for a period sufficient to affect the growth of hair and hair, the hair follicle, the size of the hair shaft, as well as the pigmentation of the hair shaft. Said topically active compositions, in addition can be combined with other active ingredients including, but not limited to, isoflavones that occur in synthetic or natural form, to improve the desired effects on hair growth and pigmentation. The compositions and methods of this invention provide a unique and convenient means to retard hair and hair growth, reduce hair follicle, hair shaft size, as well as pigmentation of the hair shaft, using serine protease inhibitors, botanical extracts of the family of legumes, and in particular, but not limited to, soy milk, which contains an inhibitory activity of serine protease and its combinations with isoflavones.

BRIEF DESCRIPTION OF THE DRAWINGS The file of this patent contains at least one drawing that has been made in color. Copies of this patent with the color drawing (s) will be provided through the Patent and Trademark Office upon request and payment of the necessary fees. The invention will be fully understood and the additional advantages will be apparent when reference is made to the following detailed description of the invention and the accompanying drawings in which: Figure 1: A photograph of the hair of a C57BI / 6 control and milk treated mouse of soybean (high amplification). Figure 2: Histological sections of hair follicles of control mouse C57BI / 6 and treated with soy milk on the fourth day of the hair cycle. Figure 3: Histological sections of mouse hair follicles C57BI / 6 control and treated with soy milk on the seventh day of the hair cycle, high and low amplifications. Figure 4: Histological sections of hair follicles of control mouse C57BI / 6 and treated with soy milk on day 18 of the hair cycle. Figure 5: Histological sections of mouse hair follicles C57BI / 6 control and treated with soy milk on day 21 of the hair cycle.

Figure 6: Histological sections of mouse hair follicles C3H control, treated with soy milk and treated with soy-derived proteins (high amplification) on day seven of the hair cycle. Figure 7: Histological sections of mouse hair follicles C57BI / 6 treated with proteins with control, soy milk and soy-derived proteins (reduced amplitude) on day seven of the hair cycle. Figure 8: Histological sections of hair follicles of control C3H mouse and treated with soybean on day 27 of the hair cycle. Figure 9: A photograph of the C3H control mouse hair and treated with soy-derived proteins (high amplification). Figure 10: Histological sections of hair follicles of control mouse C57B1 / 6 and treated with proteins derived from soybean (high amplification) at day eight of the hair cycle. Figure 11: A graph showing the inhibitory activity of trypsin in soy milk. Figure 12: Western blot of C57B1 / 6 mouse skins throughout the hair cycle, demonstrating tyrosinase and reduced TRP-1 protein levels after treatment with soy milk. Figure 13: Photographs of sides of the face of a human being, without treatment, treated with soy milk and treated daily with soy milk for four weeks. Figure 14: Quantitative analysis of the dimensions of the hair follicle with and without soy milk treatment.

Figure 15: Photographs of hair on a human's leg after five weeks of treatment with soy milk on one leg. Figure 16: Photographs of the hair of a control C57B1 / 6 mouse and treated with soy milk and soy milk enriched with isoflavones (high amplification). Figure 17: Histological sections of C57B1 / 6 control mouse skins, treated with soy milk, with soy milk enriched with isoflavones at day fifteen of the hair cycle, documenting the thickness and color of the hair stems. Figure 18: Photograph of C57B1 / 6 mouse hair after three weeks of treatment with various formulations of soy milk and soflavones.

DETAILED DESCRIPTION OF THE PREFERRED MODALITIES As used herein, "mammal" shall mean any member of "the higher-class vertebrate animals comprising the Mammalia class", as defined in the Webeter's Medical Desk Dictionary Dictionary 407 (1986), and includes, but is not limited to, to the human beings. As used herein "(%, w / v)" will mean grams of a given component per 100 ml of the total composition. Topically active agents for use in the composition of the present invention include protease inhibitors and natural plant extracts having protease inhibitory activity and mixtures thereof. Preferred protease inhibitors are inhibitors of serine protease and in particular the Trypsin Soy Inhibitor ("STI") and the Bowman Birk Inhibitor derived from soybeans ("BBI"). Preferred botanical extracts belong to the family of legumes and in particular soy extracts, such as soy milk. Preferably, the protease inhibitors are present in an amount, based on the total volume of the composition of the present invention, of from about 0.0001% (w / v) to about 20% (w / v), and most preferably around from 0.001% (w / v) to approximately 5% (w / v). Preferably, botanical aqueous extracts such as soy milk are present in an amount of 10-99% (v / v), and most preferably 50-99% (v / v). We have unexpectedly discovered that when the topically active agents, as described above and in particular the formulations containing soy and soy milk, are enriched with isoflavones, and in particular isoflavones derived from soy, it improves the inhibitory effect on hair growth. and hair, the dimensions of the hair and the pigmentation of the hair. Preferably, isoflavones are present in aqueous botanical extracts such as soy milk in an amount of 0.000005-15% (v / v), and most preferably 0.00001-10% (v / v). If the delivery parameters of the topically active pharmaceutical or cosmetic agent so require, the topically active composition of the present invention may further consist of a pharmaceutical or cosmetically acceptable carrier capable of functioning as a delivery system to allow the penetration of the topically active agent. in the hair follicle and on the skin. The pharmaceutical or cosmetic composition can optionally be combined with other ingredients such as moisturizers, cosmetic adjuvants, antioxidants, depigmenting agents, anti-aging agents, hair removal agents, hair styling agents, surfactants with sunscreen, foaming agents, conditioners, humectants, fragrances, colorants, viscosity increasing agents, pH regulating agents, preservatives and their like, as well as mixtures thereof. These will be combined in an amount that will not affect the inhibitory activity of serine protease, to produce cosmetic or pharmaceutical products such as, not exclusively, essences, creams, lotions, pastes, gels, powders, patches or injectable solutions and the like for reduction of hair growth, hair size and hair pigmentation. The compositions of this invention can be applied before, or in conjunction with, or after other ingredients or ve compositions to increase their effect. For example, the compositions of this invention may be applied together with one or more products, the purpose of which is to facilitate the removal of hair to effectively remove hair, reduce hair visibility, improve hair and hair style, or improve hair management. same. The compositions of this invention can be applied topically before, during or after hair removal. They can be applied topically together with one or more of the agents of the following group: depilatory agents, shampoo, hair conditioner, styling gel, hair care products, wax products, shaving products, hair removal products, products for after shaving, deodorants, antiperspirants, electrolysis, removal of hair by laser beam, removal of hair induced by light, masks or bath additives. The compositions of this invention can be applied daily for at least four to eight weeks, whereby an effect on the appearance of hair and hair should be observed. The application can be continued as desired to maintain the condition of the hair and hair. Daily application to the face can mitigate the condition of pseudo-folliculitis of the beard or hirsutism; The application to the axillary area can reduce the growth of hair under the arms and the application to the scalp and hair can help for the handling and styling of African hair. The composition topically active pharmaceutical or cosmetic to be applied in an effective amount for the changes arising in hair growth of a mammal hair follicle, the hair shaft size and pigmentation of the hair shaft. As used herein, "effective amount" means an amount sufficient to cover the region of the surface of the skin where a delay in hair growth is desired, in the pigmentation of the hair, as well as the reduction in the size of the hair. hair shaft. Preferably, the composition is applied to the skin surface so, based on every square cm of skin surface, from about 2 ul / cm2 to about 500 .mu.l / cm2 of topically active agent is present when you want a delay in hair growth, hair size and hair pigmentation. We have unexpectedly found that when topically active agents, such as soy milk, or milk enriched with isoflavones soy, are applied topically to the skin of an animal, a significant delay in hair growth, hair follicle and size is achieved of the hair shaft; as well as in the pigmentation of the hair shaft. It is further believed that as the hair growth cycle for humans is often lower than that of mice, it is likely that the hair growth delay in humans would be considerably prolonging in mice. The invention that is illustratively described herein may suitably be practiced in the absence of any component, ingredient, or step not specifically described therein. Various examples are set forth below to further illustrate the nature of the invention and the manner of carrying it out. However, the invention should not be considered as limited to the details below.

EXAMPLES EXAMPLE 1 Depilation of test subjects in a system of mice C57BI / 6 or C3H mice (male and female) were obtained in Charles River (Kingston, NY), aged 8-10 weeks and were in the resting (telogen) phase of their respective hair cycle. Hair growth was induced by waxing the hair of the back of each respective animal according to the procedure established in Stenn, et al., "Glucocorticoid Effect on Hair Growth Initiation: A Reconsideration", "Reconsideration on the glucocorticoid effect in the beginning of hair growth ", 6 Skin Pharmacol., 125-134 (1993). In C57B1 / 6 and C3H mice, 8-10 weeks of age; The growth phase (anagen) starts synchronously in all hair follicles at the time of depilation. As illustrated in table 1, the following observations were noted at the induction site: TABLE 1 Observations on the induction site As shown in table 1, the hair growth was visible several days after the hair removal as the pink skin of the animal began to darken. This is due to hair pigmentation in the stem since the C57BI / 6 and C3H mice contained melanocytes only in the hair follicles, and not in the epidermis of the back. A similar hair growth pattern was documented, in the international application No. PCT / US97 / 11033, when chemical depilation was developed using commercially available products. As the murine hair cycle varies not only among the strains, but also among the individual animals, the state of the hair cycle in each animal under study was analyzed. A 2 cm by 1 cm sample of skin from each mouse was isolated with scissors, fixed with a 10% pH regulated formalin solution, with a pH of about 6.9-7.1 at 25 ° C (Stephens Scientific), and then formed in a paraffin block according to well-known procedures. The block was cut with microtome, and sections were stained with H & E or Fontana-Mason dye. The sections were examined histologically to verify the phase of the hair cycle, the size of the hair follicle, the hair shaft, as well as the level of pigmentation of the hair, using procedures well known in the art. The length of the hair was visually assessed, and by using a low amplification dissection microscope (x8). This example, as well as that described in our international application No. PCT / US 97/11033, shows that the hair growth cycle for C57BI / 6 and C3H mice was approximately an average of 25 days and reports similar development timing of Stem and hair follicle without considering the method used for hair removal.

EXAMPLE 2 Preparation of soy milk and soy milk formulations One way to make soy milk is to soak soybeans in deionized or purified water for several hours, and grind them after they are completely hydrated, with the addition of small amounts of water.

(The shredding procedure allows the extraction of soy milk). After harvesting, the soy milk can be filtered to remove any residual part of the seed shell. The soy milk used in the formulations described below can be fresh soy milk, as described above or it can be made from soybean powder and water. Soybean powder is milled from soybeans and can also be lyophilized, spray-dried or freeze-dried and the resulting soy milk can or can not be filtered. Said prepared soy milk can have from about 1 to about 90% by weight of dry soybean powder. Another example is the use of soy milk powder, made from freeze-dried soya milk, spray-dried or freeze-dried, with the addition of water and finished with or without filtration or homogenization. Other soy extraction methods can also be used to create the active ingredients in the formulations described below. For example, the active ingredients can be extracted from crushed soybeans using ethanol / water mixtures, followed by removing the ethanol from the extract, in such a way that the inhibitory activity of the serine protease of the soybean is maintained, and preferably, that the protein STI remain intact. The soy products useful in this invention can be produced from any soybean species, regardless of their geographical origin, sun exposure, harvest time and the like. However, specific classes, geographic origins or growth conditions may be preferred. For example, they may be preferred, but not limited to soybean classes particularly rich in their content of Trypsin Soy Inhibitor (STI) or in isoflavone content, or growth conditions that result in enrichment of STI or isoflavone in the seed. . It should be noted that the soy products useful in the compositions of this invention have a distinctive odor, which may be tolerable in some crops, but which is not desirable in others. If necessary, the odor of the compositions of this invention can be reduced by using soy products derived from specific classes known to produce seeds with reduced odor, including but not limited to lipoxygenase-2 deficient seeds and those having a profile. of modified sugar and the like. A process for reducing oxygen levels in the formulation can also reduce odor. You can also use various disguising agents or fragrances to disguise the smell. The compositions of this invention may further comprise surfactants, humidifiers, humectants, conditioners, fragrances, colorants, preservatives, antioxidants, depigmentation agents, hair removal agents, anti-aging agents, sunscreens, foaming agents, cosmetic adjuvants, hair regulating agents. pH or mixtures thereof. The compositions of this invention can be left on the skin for a period sufficient to effect changes. For example, the compositions of this invention can be applied to the daily treatment of the skin for at least about four weeks, preferably, the composition should be applied daily for at least eight weeks.

Another method according to this invention is a method to reduce or avoid pseudo-folliculitis of the beard. The daily application of the compositions of this invention can reduce or avoid this condition. The compositions of this invention can also be applied daily to the armpit area to reduce hair growth. In addition, the compositions of this invention can be applied daily to the scalp to stylize and improve the handling of African-type hair. As shown in our patent application of E.U.A. Copendent (Case of Attorney No. JBP 430), you can use many formulations based on soy milk to reduce pigmentation. All these formulations can also be used to reduce hair growth. Some particularly preferred examples of soy milk formulations and soy milk formulations containing isoflavones are shown below in Table 2. An example for the preparation of isoflavones which can be used in this invention is Flavosterone SE from Ichimaru, Japan , which contains approximately 0.1% pure isoflavones. In all these formulations, you can replace soy milk with the right amounts of soy powder or soy milk powder and water.

TABLE 2 Formulations for soy milk essence EXAMPLE 3 Preparation of topically active compositions containing protease inhibitors derived from soy Soybean trypsin inhibitor (STI) and Browman-Birk inhibitor (BBI), from Sigma-Aldrich Corporation, were mixed in 0.1 M phosphate buffered saline (PBS, Gibco-BRL, Gaithersburg, MA), pH 7.4, in concentrations of 1% to 0.001% (w / v). Then four volumes of the resulting solutions were mixed with 1 volume of liposome vehicle (100 mg / ml), which was prepared by the methods described in Niemiec et al, in order to produce the topically active composition. Nonionic liposome preparations, such as those described in Niemiec et al., "Influence of Nonionic Liposomal Composition On Topical Delivery of Peptide Drugs Into Pilosebaceous Units: An In Vivo Study Using the Hamster Ear Model," 12 Pharm. Res. 1184-88 (1995) ("Niemiec"), which is incorporated herein by reference in its entirety, are known in the art and are described in our patent application of E.U.A. (Case of Attorney No. JBP 430). The GDL liposomes were prepared as set forth in Niemiec, et al., With the exception of the following changes: the non-ionic liposomal formulation contained glycerol dilaurate (Emulsynt GDL, ISP Van Dyk) / cholesterol (Croda) / 10-stearyl ether polyoxyethylene (Brij76, ICI) / polyoxyethylene 9-lauryl ether in a ratio of 37.5: 12.5: 33.3: 16.7. The pH buffer PBS or Hepes, 0.05M, pH 7.4 (Gibco-BRL from Gaithersburg, MD) was used as the aqueous phase in the preparation of the liposomes.

EXAMPLE 4 Soy milk retards hair growth and reduces the size of hair shaft and hair follicle and hair shaft pigmentation C57BI / 6 mice were induced for a new hair cycle as described in example 1 and treated daily with soy milk. The animals were observed daily for their hair growth pattern and skin biopsies were taken at important points of the hair cycle time. As a result of treatment with soy milk, the hair growth of the treated mice was retarded and their fur was visibly thinner and softer to the touch. The treated mice showed no skin darkening on days 7-8 of the hair cycle as expected, and hair stems were not visible on days 11-12 as in the control animals. On average, the hair cycle of the mice treated with soy milk was delayed on days 3-6. Figure 1 is a photograph of the fur of the mice, showing the difference in appearance, color, size and thickness of the hair following the treatment with soy milk. Histological examination of the skin samples confirmed these observations. As shown in figure 2, by Fontana-Mason staining (F & amp; amp;M), on day four of the hair cycle, the untreated hair follicle develops fully as expected, containing all cell layers and pigment deposition. In contrast, the sample treated with soy milk, (shown in the same amplification), shows a smaller and less fully developed hair follicle, without pigment deposition. Figure 3 shows two series of histological sections stained with F &M, higher and lower amplification. These sections correspond to day 7 of the hair cycle. The upper panel shows that the skin treated with soy milk has smaller, shorter and less pigmented hair follicles than the untreated control. The lower panel shows a higher amplification of the follicles, also demonstrating the difference in size and pigmentation of hair shaft and follicle following the treatment with soy milk. Figure 4 shows a low amplification of sections of skin stained with F &M on day 18 of the hair cycle. In this amplification, it is obvious that treatment with soy milk results in a reduced size of hair follicle, which leads to a reduced length and thickness of the hair shaft and to a reduced total pigment deposition within the treated follicles. Figure 5 shows sections of skin on day 21 of the hair cycle, with two amplifications. The upper panel shows that the control animals were in the catagen stage, when the hair follicles are receding. On the other hand, follicles treated with soy milk, have completed the stage of catagen, since they are shown in telogen, the resting stage. This indicates that not only was the hair cycle retarded following the treatment with soy milk, but it also ended prematurely. The lower panel shows the follicle control in catagen and the follicle treated with shorter soy milk, in telogen (rest) using higher amplification.

EXAMPLE 5 Effects of soy milk on hair growth, size and pigmentation are reproducible in C3H mice In order to verify that the effect of soy milk on hair growth is not specific for C57BI / 6 mice, the experiment described in example 4 was repeated using brown hair mice (Agouti) C3H. The results of these experiments were similar both visually and histologically. Soy milk retarded hair growth and reduced the size of hair shaft and follicle and pigment deposition in C3H mice. Histological analysis confirmed these visual observations. As shown in the upper panel of Figure 6, using F &M staining, on day seven of the hair cycle, the follicles treated with soy milk are smaller and accumulate less pigment than the untreated controls. The upper panel of Figure 7 (F &M staining) shows a lower amplification of the same skin sections, demonstrating the thinner and less pigmented follicles following the treatment with soy milk. Figure 8 shows sections of skin stained with F &M on day 21 of the hair cycle. As shown for C57BI / 6 mice following soy milk treatment, the hair cycle ends prematurely. The follicles treated with soy milk are in the resting stage, while the untreated control follicles are still in catagen.

EXAMPLE 6 The effects of soy milk and soybean derived from serine protease inhibitors on hair growth, size and pigmentation In search of a mechanism to explain the effect of soy milk on hair growth, the effect of serine protease inhibitors derived from soy milk, STI and BBI was tested. It has been shown above that these proteins induce depigmentation in the skin, affecting the PAR-2 pathway (patent application of E.U.A., Case No. JBP 430). The experiments described in example 4 were repeated using STI, BBI and soy milk. STI and BBI were used in a PBS-liposome vehicle as described in example 3. Visual observations throughout the hair cycle confirmed that both STI and BBI could delay hair growth and reduce the size of the follicle and stem. hairy, similar to that of soy milk (see hair photos in figure 9). Using high concentrations of STI or BBI, the effect on hair growth and pigmentation was substantial. Histological analyzes confirmed this discovery. As shown in figure 6, day seven of the hair cycle, 1% of STI and 1% of BBI reduced the size of the hair follicle and stem, as well as the hair shaft pigmentation in C3H mice. Figure 7 shows sections of smaller extension of the same day in the hair cycle, demonstrating smaller hair follicles and stems, as well as reduced pigmentation, in relation to the untreated control, with treatment with soy milk, STI or BBI. Figure 10 shows that STI and BBI have the same effect in C57B1 / 6 mice as well, demonstrating smaller and less pigmented follicles. Taken together, this example shows that STI and BBI are serine protease inhibitors derived from soy, found in soy milk, which could delay hair growth, reduce the size of the hair follicle and stem, and reduce hair pigmentation. STI and BBI could represent a part of soy milk ingredients that affect hair growth. To support the hypothesis that STI and BBI in soy milk are involved in the hair growth effects described above, soy milk was tested to verify its inhibitory serine protease activity. An enzymatic test was carried out using "Enzchek", a fluorescent protease digestion test system made by Molecular Probes of Eugene, OR. Using 100 units of trypsin (from Sigma chemicals, San Luis MO) the test system produced a fluorescence reading of approximately 1100 units. This reaction was inhibited with increasingly high concentrations of STI, as expected from a known trypsin inhibitor. Serial dilutions of soy milk were tested in this test, and were found to inhibit trypsin activity. As shown in Figure 11, soy milk exerts trypsin inhibitory activity similar to about 0.2% pure STI. This suggests that soy milk could exert its effect on hair growth, at least in part, through STI and BBI.

EXAMPLE 7 Soy milk induces changes in the expression of tyrosinase and TRP-1 protein The histological analyzes of skin samples treated with soy milk described in the previous examples shows a dramatic reduction in pigment deposition within the hair follicle. To further understand the mechanism of depigmentation induced by soy milk, tyrosinase was studied, the key enzyme in melanogenesis and tyrosinase-related protein 1 (TRP-1), the enzyme that stabilizes tyrosinase. Mice C57B1 / 6 and C3H were treated as described above, and samples were taken throughout the study for protein analysis. Protein extraction and Western blot analysis were carried out using standard procedures, such as that described in Current Protocols in Cell Biology, Edited by Juan S. Bonifacino et al. Title 6: Electrophoresis and Immunoblotting. Copyright 1999 by John Wiley & Sons, Inc., which is incorporated herein by reference in its entirety. An example of one of these studies is shown in Figure 12. Equal amounts of proteins extracted from skin were tested with the anti-tyrosinase antibody "anti PEP1", and with the anti-TRP-1 antibody "anti PEP7" which were describe in Jiménez, M., Kameyama, JK., Maloy, WL, Tomita Y. and Hearing, V. Mammalian tyrosinase: biosynthesis, processing and modulation by melanocite stimulating hormone. Proc. Nati Acad. Sci. USA (1988), 85: 3830-34, and Jiménez, M., K., Maloy, WL, and Hearing, V. Specific dentification of an authentic tyrosinase clone. J. Biol. Chem. (1989) 264: 3397-3403, which are incorporated herein by reference in their entirety. As shown in Figure 12, the expression of tyrosinase and TRP-1 proteins is dramatically affected by treatment with soy milk. The levels of tyrosinase and TRP-1 are reduced, and the duration of expression is shortened. These two factors affect the general pigmentation of hair, which decreases due to the reduced level and shorter duration of melanogenesis.

EXAMPLE 8 Soy milk reduces the length and thickness of human facial hair A male individual with dark facial hair shaving daily treated the right side of his face with soy milk, immediately after shaving, for five weeks. By the third week, and most notably by the fourth week, the hair on the treated side was visually lighter and felt softer to the touch. Digital photos of different magnifications were taken throughout the treatment period using Hi-Scope. These photos clearly demonstrate the reduced size and thickness of the hair stems on the treated side. An example of such photos is shown in Figure 13, demonstrating the difference in the thickness and density of the hair shaft at four weeks of treatment. Since both sides of the face were shaved at the same time, and that the photos of both sides were taken at the same time, the difference in facial hair length indicates a slower growth rate in the treated area. Figure 14 shows a computerized image analysis of the length, thickness and total area of facial hair, after four weeks of treatment with soy milk. All images were analyzed with Image Pro Plus 3.0 software (Media Cibemetics, Silver Spring, MD). The data are presented as an average of 180 hair stems on each side of the face, with standard deviation (SigmaPlot® 5.0, SPSS Science, Chicago, IL). The statistical analysis was carried out using SigmaStat® 2.0 software (SPSS Science), demonstrating a statistically significant difference in all parameters measured, after soybean milk treatment.

EXAMPLE 9 Soy milk reduces the length and thickness of human leg hair Hair was waxed from two symmetrical areas of the middle part of the legs, below the knee, in an individual. One leg was treated with soy milk daily, for four weeks. Visual observations indicate slower hair growth in the treated area. The hair stems were reduced in number and were shorter and thinner than those of the untreated site, as shown in Figure 15. These observations further confirm the effect of soy milk on hair growth. Examples 8 and 9 confirm together that the effect of soy milk on the growth of human hair is not related to the method of hair removal with the part of the body that is being treated.

EXAMPLE 10 Soy milk formulations enriched with isoflavones are preferred over soy milk formulations to reduce hair growth and pigmentation.

The experiments described in Example 4 were repeated, using the two formulations described in Table 2 above. Essence of soy milk 23, which is a formulation based on soy milk, and soy milk essence 30 which is identical to the soy milk essence 23 except for the addition of 5% of an isoflavone extract to the 0.1%. As shown in Figure 16, mice treated with soy milk essence 23 showed reduced hair growth and improved appearance thereof. This effect was more pronounced with the use of soy milk essence 30, demonstrating that soy milk formulations enriched with soflavone are superior to those of soy milk formulations to reduce hair growth. Figure 17 shows sections of histological skin hair from the treated mice, on day 15 of the treatment. The hair stems documented in these sections clearly demonstrate the dimensions of the hair shaft, the reduced level of pigmentation within the hair shaft, and the increased softness of the hair shaft after treatments with soy milk essence.

EXAMPLE 11 Soy milk formulations enhanced with isoflavones are preferred over isoflavone formulations which are effective in reducing hair growth and pigmentation.

The experiments described in Example 4 were repeated, using the formulations described in Table 2 above, of soy milk essence with or without increasing concentrations of soflavones. These soy essence formulations were compared with similar formulations in which the soy milk component was only replaced with water. These three sets of formulations (soy essence, isoflavones, soy essence containing additional soflavones) were prepared to test the possibility that isoflavones might be sufficient for the observed effect on hair growth. Figure 18 shows CS7B1 / 6 mouse hair after three weeks of topical treatment, as described in Figure 4. Both the untreated control mice and the placebo treated mice have long, less "ordered" hair. The soy milk essence 23 reduces hair growth and leads to a better appearance, as described earlier in this application. Soy milk formulations containing 1, 5 and 10% isoflavones containing 0.1% extract result in a superior effect on hair growth. However, formulations that contain isoflavones but not soy milk demonstrate a milder effect, and not as superior as when combined with soy milk. This example demonstrates that formulations of soy milk containing isoflavone could reduce hair growth. This example further demonstrates that formulations of soy milk containing isoflavones reduce hair growth to a greater degree than formulations containing soflavones alone.

EXAMPLE 12 Soy essence formulations affect the growth of human hair The efficacy and irritation potentials of soy milk essence 23 and 30 compared to a placebo formulation were examined in a four-week placebo-controlled blinded trial with twelve premenopausal women panelists aged 29 to 45 using evaluations made by the researcher of the study, self-evaluation by panelists and Hi-scope image analysis. The panelists signed an Informed Consent, were instructed about the procedures and expectations of the study and asked to shave that night. At the baseline visit the next day, two test lotions were distributed to each panelist (day 1), a placebo lotion and soy milk essence 23 or 30. The lotions were randomly assigned to the right leg or the left leg. The test lotions and placebo were used on the respective legs for the duration of the study without the application of other lotions on the lower parts of the legs. The panelists were instructed to apply the test lotions twice a day, in the mornings and afternoons, and were also instructed to refrain from shaving the lower legs until after each weekly evaluation. On the evaluation days, the investigator visually reviewed the legs of the panelists looking for any clinical sign of irritation and compared the legs to verify attenuation in hair growth. The self-assessment questionnaires were completed by panelists at each evaluation time point (weeks 1, 2, 3 and 4). In addition, Hi-scope images (2.5 cm in diameter for each image, KH-2400R, Hirox) were obtained at each time point using an MX-MACROZ lens (Hirox). No panelist left the study for reasons related to the product. No signs of irritation were observed in any of the study panelists at any point in time, nor was any irritation reported when they were self-evaluated by the panelists at any time during the study. For the purpose of hair counts, all hair, including "bristles" was counted in the 2.5 cm field given to each panelist at each time point. The results showed a decrease in lower leg hair counts at week 5 for soy milk essence 30 and at week 4 for soy milk essence 23. The legs treated with placebo showed no change at all. the average leg hair counts throughout the study, although the standard deviations were large. The rate of growth was calculated by dividing the length of time (in days) since the panelist was last shaved, by the average length of hairs on the leg for that panelist, which was calculated from the Hi-scope images used Image Pro Plus analysis for each panelist at each test site. The results are documented in Table 3, showing that both treatments with soy milk essence 23 and 30 resulted in a reduced hair growth rate compared to placebo.

TABLE 3 Average speeds of hair growth in legs after treatment with soy milk essence or placebo • The results of the self-assessment questionnaires of the 15 panelists showed that the panelists felt that the test lotions attenuated the hair growth and softened the sensation of the hair of the legs, in comparison with the placebo lotion. The panelists felt that the hair felt less thick and less bristly. Most of the panelists believed that the test lotions decreased the growth of the hair on the leg or altered the hair texture so that it felt softer and less thick. The H¡ scope analysis also showed that hair regrowth after treatment with soy milk essence 23 or 30 seemed to grow in the same direction and was more uniform in shape, texture and appearance. On the contrary, the hair that grew back on the legs treated with placebo grew in different directions, with differences in length, angle of growth and thickness. This example clearly demonstrates the effect of soy milk formulations to retard and reduce hair growth, and allow hair growth to be smoother, thinner and more manageable and directionally organized.

Claims (52)

NOVELTY OF THE INVENTION CLAIMS

1. - A method for effecting changes in the appearance of mammalian hair, hair growth, hair pigmentation and hair follicle size and hair shaft, comprising topically applying to the skin of a mammal an effective amount of a topically active composition comprising one or more compounds derived from one or more of the botanical families Leguminosae, Solanaceae, Gramineae and Cucurbitaceae.

2. The method according to claim 1, further characterized in that said extract contains at least one inhibitory activity of serine protease.

3. The method according to claim 2, further characterized in that at least one serine protease inhibitor active agent is a serine protease inhibitor.

4. The method according to claim 3, further characterized in that at least one serine protease inhibitor is present in an amount, based on the total volume of the topically active composition, of about 0.0001% (w / v) ) to 20% (p / v).

5. The method according to claim 4, further characterized in that at least one serine protease inhibitor is present in an amount, based on the total volume of the topically active composition, of about 0.001% (w / v) ) at approximately 5% (w / v).

6. The method according to claim 1, further characterized in that one of said changes is the growth retardation of hair, reduction in the size of hair follicle and hair shaft and reduced hair pigmentation.

7. The method according to claim 1, further characterized in that said topically active composition additionally comprises a pharmaceutical or cosmetically acceptable vehicle.

8. The method according to claim 1, further characterized in that said topically active composition additionally comprises one or more soflavones.

9. The method according to claim 8, further characterized in that said topically active composition additionally comprises a pharmaceutical or cosmetically acceptable vehicle.

10. The method according to claim 1, further characterized in that said topically active composition additionally comprises natural extracts containing one or more soflavones.

11. The method according to claim 1, further characterized in that said composition is applied topically in conjunction with one or more products whose purpose is to facilitate the removal of hair or effectively remove the hair or reduce the visibility of hair or improve the style or hair management.

12. The method according to claim 1, further characterized in that said composition is applied topically before or after hair removal.

13. The method according to claim 1, further characterized in that said composition is applied topically during the removal of hair.

14. The method according to claim 1, further characterized in that said composition is applied topically in conjunction with one or more of the group consisting of: depilatory agents, shampoo, hair conditioner, styling gel, hair care products , wax-based products, shaving products, hair removal products, after-shave products, deodorants, antiperspirants, electrolysis, laser hair removal, light-induced hair removal, masks, and bath additives.

15. The method according to claim 1, further characterized in that it consists in leaving said composition on said skin for a period sufficient to effect said changes.

16. The method according to claim 1, further characterized in that said period is a daily treatment of at least about four weeks.

17. - A method according to claim 16, further characterized in that said composition is applied daily for at least eight weeks.

18. A method according to claim 1, further characterized in that said composition is applied daily to reduce or prevent Pseudofolliculitis barbae.

19. The method according to claim 1, further characterized in that said composition is applied daily in the area of the armpit to reduce hair growth.

20. The method according to claim 1, further characterized in that said composition is applied daily on the scalp to improve the style and management of African-type hair.

21. The method according to claim 1, further characterized in that said composition is added daily to a bath.

22. The method according to claim 1, further characterized in that said composition is used daily on the face or parts of the body to retard the growth of hair and reduce the visibility of hair in those suffering from hirsutism.

23. A composition for effecting changes in hair growth in mammals, hair pigmentation and hair shaft size and hair follicle, which consists of applying topically on the skin of a mammal an effective amount of a topically active composition comprising one or more compounds derived from one or more of the botanical families Leguminosae, Solanaceae, Gramineae and Cucurbitaceae.

24. The composition according to claim 23, further characterized in that the extract contains at least one inhibitory activity of serine protease.

25. The composition according to claim 24, further characterized in that at least one serine protease inhibitor active agent is present in an amount, based on the total volume of the topically active composition, of about 0.0001% (p. / v) to approximately 20% (w / v).

26. The composition according to claim 25, further characterized in that at least one protease inhibitor active agent is present in an amount, based on the total volume of the topically active composition, of about 0.001% (p / v) at approximately 5% (w / v).

27. The composition according to claim 23, further characterized in that the composition additionally comprises one or more soflavones.

28. The composition according to claim 23, further characterized in that the composition additionally comprises one or more natural extracts containing one or more isoflavones.

29. The composition according to claim 23, further characterized in that the composition additionally comprises surfactants, humectants, wetting agents, foaming agents, cosmetic adjuvants, pH regulating agents, preservatives, antioxidants, conditioners, depigmenting agents , hair removal agents, anti-aging agents, sunscreen, fragrances, dyes, or mixtures thereof.

30. The composition according to claim 23, further characterized in that said composition is in the form of liquid, cream, gel, paste, powder, essence, mousse, emulsion, film or aerosol patch.

31. The composition according to claim 23, further characterized in that said composition additionally comprises one or more products whose purpose is to facilitate the removal of hair or actually remove the hair or reduce the visibility of hair or improve hair management.

32. The composition according to claim 23, further characterized in that said composition further comprises one or more of the group consisting of: depilatory creams, shampoo, hair conditioner, styling gel, hair care products, products based Waxing, shaving products, hair removal products, after shave products, deodorants, antiperspirants, pre- or post-electrolysis products, pre- or post-laser hair removal products, hair removal products induced by pre- or post-light, masks, and bath additives.

33. - A composition according to claim 23, further characterized in that said compound is derived from legumes.

34. A composition according to claim 33, further characterized in that said compound is derived from soybean, bean and / or bean seeds.

35. A composition according to claim 34, further characterized by said compound is selected from the group consisting of soy milk, bean milk, bean milk, soy extract, bean extract, bean extract, pasta soybean, bean paste, bean paste, soybean powder, bean powder, bean powder, soy milk powder, bean milk powder and bean milk powder and mixtures thereof.

36.- A composition according to claim 35, further characterized in that said compound is a fraction of soy milk, soybean extract, soybean paste, bean milk, bean extract, bean paste, bean milk, extract of bean, bean paste, soybean powder, bean powder, bean powder, soy milk powder, bean milk powder, and bean milk powder and mixtures thereof.

37. A composition according to claim 35, further characterized in that it comprises bean milk in an amount of about 1 to about 99% by weight.

38.- A composition according to claim 35, further characterized in that it comprises soybean powder, soy milk powder or a mixture thereof in an amount of about 1 to about 99% by weight.

39. A composition according to claim 36, further characterized in that it comprises soybean trypsin inhibitor, bean trypsin inhibitor or bean trypsin inhibitor in an amount of about 0.0001 to about 20% by weight.

40.- A composition according to claim 33, further characterized in that said composition additionally comprises one or more isoflavones.

41. A composition according to claim 33, further characterized in that said composition additionally comprises one or more natural extracts containing one or more isoflavones.

42. A composition according to claim 40, further characterized in that said one or more isoflavones are present in said composition in an amount of about 0.00001 to about 0.1% by weight.

43. A composition according to claim 41, further characterized in that said one or more natural extracts contain said one or more isoflavones in said composition in an amount of about 0.00001 to about 0.1% by weight.

44. A cosmetic composition according to claim 23, further characterized in that said composition additionally comprises a cosmetically acceptable vehicle.

45. - A composition according to claim 23, further characterized in that said composition additionally comprises liposomes.

46. A composition according to claim 45, further characterized in that said composition comprises glycerol dilaurate, cholesterol, polyoxyethylene-10-stearyl ether and polyoxyethylene-9-lauryl ether.

47. A composition according to claim 23, further characterized in that said composition comprises from about 1 to about 99% bean's milk, from about 0.1 to about 20% emulsifier and preservatives in an effective amount.

48. A composition according to claim 23, further characterized in that said composition comprises from about 0.5 to about 20% soybean powder or soy milk powder, from about 0.1 to about 20% emulsifier and preservatives in an effective amount.

49. A composition according to claim 47, further characterized in that said composition further comprises one or more isoflavones in an amount of about 0.00001 to about 0.1%.

50.- A composition according to claim 47, further characterized in that said composition further comprises one or more natural extracts comprising one or more isoflavones in an amount of from about 0.0001 to about 0.1%.

51. A composition according to claim 23, further characterized in that said composition further comprises one or more soflavones in an amount of about 0.0001 to about 0.1%.

52. A composition according to claim 48, further characterized in that said composition additionally comprises one or more natural extracts comprising one or more isoflavones in a 10 amount of about 0.0005 to about 0.05%. r