KR20050019120A - Compositions and methods for softening, thinning and removing hyperkeratotic tissue - Google Patents

Abstract

Novel compositions and methods of treatment are disclosed for the treatment of mammalian keratinous tissue using proteolytic enzymes originally developed to hydrolyze food-related proteins and commonly used as food grade meat tenderizers and flavor enhancers. The protease-containing composition safely softens hyperkeratin on the skin, including calluses, nails, dry, aborted and flaky skin and keratin, with no harm to surrounding tissues by selectively hydrolyzing only keratinocytes. Paint, thin and peel. These enzymes accomplish this by breaking down only the 10-end peptide bonds of keratin proteins, which are the major protein components of keratinous skin.

Description

Compositions and methods for softening, thinning and removing hyperkeratomic tissues

The present invention relates to a composition for treating conditions associated with mammalian keratinous tissues of proteolytic enzymes developed for hydrolyzing food-related proteins and their use.

The keratinous tissue of the mammal and the conditions resulting therefrom result from abnormal processes in the skin. The upper layer of skin known as the stratum corneum consists mainly of cells, which are dehydrated keratin proteins. In a normal process called keratinization, epidermal cells are slowly pushed to the surface by the lower dermis and peel off there. When an abnormal process, i.e., when these cells are peeled off, they accumulate on the surface, forming keratinous tissue, causing discomfort. In mammals, keratinous tissue includes calluses (calks and fissures), corns (pegs), keratin (rashes), and dry skin drops and flaking. A common way to manage hyperkeratin tissue is to apply an acid in the form of a lotion or cream carrier and to wet or mechanically peel off the cream or lotion. Acids and peels tend to irritate normal epidermis and are not advantageous and wet creams are not effective in reducing hyperkeratin tissue density.

Calluses and nails, often found in feet or hands, are areas of thickened normal skin that are formed to protect the underlying bones from excessive weight, pressure, or friction. Calluses on the feet most commonly occur on the soles and heels. Calluses on the hands are typically formed on the pads of the palm, where pressure is most commonly encountered when handling tools or objects. When calluses accumulate dead cells at their epicenter, they become nails, causing painful conical keratinization. Nails are usually found between toe nodes and toes and are typically caused by hammer toes, bone spurs, or incorrect shoe wear to the extent of the skin. Calluses and nails are often cured by rubbing hard skin using pumice or other scraping instruments and applying salicylic acid, or in severe cases, bone surgery to correct lower pressure points. It is often treated by. Salicylic acid is effective in removing calluses and nails, but also has a detrimental effect on adjacent healthy tissue, causing it to peel off. Salicylic acid is also not recommended for use in elderly people with thin skin, diabetics, or those with weaker circulation or compromised immune systems, which can lead to open lesions susceptible to the acid infection.

Dry skin is a normal consequence of aging and is exacerbated by cold, dry or windy weather conditions. Treatment involves the use of humectants, including petrolatum, lanolin, urea, alpha hydroxy acids, lactic acid or ammonium lactate, after bathing to reduce abortion and moisturize the skin.

eczema

Eczema is a generic term that encompasses a variety of infectious skin conditions, including the very bleeding areas of the face, neck, cuffs, knees and ankles and dry, red, and deciduous areas that typically occur on the feet and hands. In the United States, about 15 million people suffer from eczema, and about 15% of infants have symptoms but get older. In some eczema, the skin can become thick, like leather. The common form of eczema is atopic dermatitis, whose manifestations are presumed to be due to abnormal reactions of the body's immune system or to stimulants in contact with the skin. Therapies are not yet known and the condition is not contagious. The disease can be managed by avoiding irritants or by taking medication.

Onset prevention uses a number of means, humectants and anti-itch creams designed to avoid skin irritation and prevent excessive drying, or avoid sensational foods such as sudden temperature changes, stress reduction, strong chemicals such as detergents, or wool And by limiting environmental contaminants such as mold, pollen or animal dandruff. Medications include corticosteroids, antihistamines, coultar therapy, phototherapy and in some serious cases include topical immunomodulators such as cyclosporin A and tacrolimus.

Cuticle (cuticle)

The skin covering the borders of the nails and toenails is called the cuticle. The cuticle can be cured by frequent exposure to irritants such as soaps, detergents and chemicals, or by physical abuse such as nailing, gardening without protecting the hands, or by extreme cold weather. Current methods for managing hardened or enlarged toenail cuticles are to cut out thickened or grown areas using various manicure instruments. This may have the undesirable effect of creating a new corneal layer to replace the removed portion, or in some cases exposing the dermis at risk of infection or irritation. Healthy cuticles are smooth, transparent and have a uniform contour. Damaged cuticles are thick, dry, thickened, opaque, adhered strongly to the nail plate, sometimes extending to the nail plate. A number of cuticle creams are currently used to improve damaged cuticles. These include emollients, skin humectants and acids.

Biting your nails leads to biting the cuticle. This causes tearing and tearing of the cuticle of the nail, and when peeled from the dermis, it dries to form a hard crust of the skin around the lower arch of the nail.

1 is a graph showing the effect of pH on the activity of one of the proteolytic enzymes of the present invention.

Figure 2 is a graph showing the effect of temperature on the activity of one of the proteolytic enzymes of the present invention.

3 is a graph showing the productivity of one of the proteolytic enzymes of the present invention.

Figure 4 is a graph showing the effect of pH on the activity of one of the proteolytic enzymes of the present invention.

5 is a graph showing the effect of temperature on the activity of one of the proteolytic enzymes of the present invention.

6 is a plantar photograph of a patient prior to treatment in accordance with the present invention.

 7 is a photograph of the sole of the patient according to FIG. 6 after treatment in accordance with the present invention.

8 is a heel photograph of a patient before treatment in accordance with the present invention.

9 is a photograph of a heel of the patient according to FIG. 8 after treatment in accordance with the present invention.

Summary of the Invention

The present invention relates to a composition for the treatment of mammalian keratinous tissue and its use using proteolytic enzymes originally developed to hydrolyze food-related proteins and commonly used as meat tenderizers and flavor enhancers for food. Surprisingly, the inventors have found that the novel composition containing the proteolytic enzymes safely softens, thins and exfoliates the keratin on skin, including callus, nails, dry, abortion and flaky skin and keratin. In particular, when present in a carrier suitable for use on the skin and with or without obstruction on the skin, such proteins and proteases selectively degrade hyperkeratin tissues, resulting in non-hyperkeratin tissues. It can be softened, softened, thinned and peeled off without any harm. These enzymes accomplish this by breaking down only the 10-end peptide bonds of keratin proteins, which are the major protein components of keratinous skin. These substances have minimal effect on normal skin because healthy epidermis lacks keratin other than that present in the microlayers of dead skin cells associated with normal peeling. An example of a proteinase enzyme suitable for the above uses is Subtilisin Carlsberg (made by Novo Corporation) prepared for Alcalase® foods. Another example is a fungal protease / peptidase complex produced by submerged fermentation of selected strains of Aspergilus oryzae marketed by Novo Corporation as Falvourzyme ^™ flavor enhancer.

Enzymes that break down proteins are widely distributed in nature. For example, US Pat. No. 5,213,978 discloses extracellular proteases obtained from Micrococcus sedentarius capable of dissolving human callus material. However, the present invention utilizes proteolytic enzymes that selectively degrade mammalian keratinous tissue. Tests conducted under bilateral, placebo control, double blind conditions show a 50% reduction in hardness and density when applied to the callus of the foot for 14 days without any irritation to adjacent normal skin.

Selective degradation of calluses, nails and keratin is because normal skin lacks proteins that contain 10-end peptide bonds that are degraded by proteolytic enzymes. Articles currently used to treat calluses, nails, keratin and dry volumes generally contain acids that can irritate normal skin and therefore lead to burnout of healthy tissue, which can lead to infection.

Surprisingly, the combinations according to the invention are effective in controlling eczema symptoms. In a preferred embodiment at least one protease of the present invention is provided in a mineral oil base and the combination is applied topically to the skin area exhibiting eczema. The protease is believed to degrade infected tissues including abortion and thickened or leathery epidermis.

The present invention is useful for removing or exfoliating dead cells from the surface of the skin, known as the stratum corneum, and providing a softer tissue to the skin, but especially with a night ointment to prevent the appearance of wrinkles and lines and to reduce the appearance of fine lines and wrinkles. It is not limited to facial skin when used. In a preferred embodiment, the present invention includes a mineral oil base that acts as a carrier of the enzyme / protease mixture and acts as a moisture barrier to keep moisture on the face and prevent drying. Ointment can be used nightly or once a week depending on age and dermatitis condition.

The protease mineral oil ointment formulations described herein slowly thin and soften the dryness, hyper- and cured keratin of nails and toenails. Dead cells formed on the cuticle of the nail can be removed within one or two days of applying the combination of the present invention. Thinning and softening the cuticles covered or grown from the nail plate and then peeling off over time provides better appearance and better protection around the nail plate. As a result, methods such as cutting with a manicure tool or biting the cuticle portion to remove loose skin are not appropriate, resulting in cosmetic problems or infection problems.

Purpose of the Invention

The present invention provides several unique advantages over the prior art.

The present invention is intended primarily for the hydrolysis of food-related proteins, containing proteinases that soften, thin and exfoliate hyperkeratoid formation on skin, including callus, nails, dry, ablative / flake skin and keratin. It provides a novel composition for the treatment of keratinous tissue in a mammal containing the known known proteolytic enzymes.

The present invention also provides a method of administering a novel composition containing proteolytic enzymes for the treatment of mammal's keratinous tissue.

The present invention also provides a novel composition for exfoliating mammal's keratinous tissue at body temperature and neutral pH.

The invention also provides dermatological use of proteolytic enzymes in a suitable carrier, characterized in that the proteolytic enzyme selectively degrades the keratinous tissue to soften, smooth, thin and peel off without harming the non-keratinous tissue. The present invention provides a method of regularly applying and administering to skin with or without occlusion.

The present invention can be understood in more detail with reference to the detailed description and drawings of the present invention shown below.

The novel composition of the present invention comprises a proteinase or proteolytic enzyme in a carrier suitable for dermatological use. An example of a proteolytic enzyme suitable for this use is Subtilisin Carlsberg , manufactured by Novo Corporation for Alcalase® foods. Subtilisin Carlsberg is an endopeptidase produced by submerged fermentation of selected strains of Bacillus licheniformis . The main enzyme component, subtilisin A (= subtilisin Carlsberg), is an endoproteinase widely described in the literature. Table 1 summarizes the biochemical properties of proteinases in alcalase. Optimal conditions for alcalase range from 55 ° C. (131 ° F.) to 70 ° C. (158 ° F.), depending on the type of substrate, and have a pH value of 6.5 to 8.5.

Alcalase food use is expressed in Anson units per gram (AU / g).

Alcalase 2.4L ........... Identified Activity: 2.4 AU / g

Alcalase 2.4L is a dark brown liquid and easily soluble in water at all concentrations. Colors can vary from batch to batch and color intensity is not an indication of product intensity.

activation

By analyzing alcalase at various pH values using a modified Anson / hemoglobin method, its optimal activity was found at pH 6.5 to 8.5 (FIG. 1). Similarly, under assay conditions, alkalase shows good activity (about 40%) at normal body temperature (37 ° C.) in humans. Alcalase is at an optimum temperature near 60 ° C. (140 ° F.) (FIG. 2).

3 shows the productivity of alcalase at various temperatures on matrix whey protein and soy protein. The productivity is depicted relative. DH is defined as the ratio of degraded peptide bonds.

Alkalase can be inactivated within 30 minutes at a temperature of 50 ° C. (122 ° F.) or higher at pH 4 and within 10 minutes at a temperature of 85 ° C. (185 ° F.) or higher at pH 8. However, this inactivation is very dependent on the substrate (substrate concentration, pH).

Another example of a suitable proteolytic enzyme is one sold under the name Flavorzyme as a flavor enhancer. Flavourzyme, marketed by Novo Corporation, is a fungal protease / peptidase complex produced by submerged fermentation of selected strains of Aspergillus oryzae . Such proteolytic enzymes have both endoprotease and exopeptidase activity. Flavorzyme is currently used as a food processing aid to eliminate taste bitter protein hydrolysates and to increase flavor production by increasing hydrolysis of food proteins.

The optimal pH for the enzyme complex is in the range of 5.0-7.0. The optimal pH for exopeptidase is about 7.0 as determined by application experiments. A suitable pH for eliminating bitter taste is about 7.0. The optimum temperature for the enzyme complex as well as exopeptidase is about 50 ° C. (122 ° F.).

Flavorzyme is a standardized leucine amino peptidase unit (LAPU / g) per gram.

Flavorzyme 500 MG ......... Identified Active: 500 LAPU / g

Flavorzyme 500 L .......... Identified Active: 500 LAPU / g

One LAPU is the amount of enzyme that hydrolyzes 1 pmol of L-leucine-p-nitroanilide per minute. Flavorzyme is available as Flavourzyme 500 L, a liquid product, and Flavorzyme 500 MG, a brown, free-flowing, non-dusty microgranule granulated on NaCl. Color is batch dependent and color intensity does not indicate product intensity. Flavorzyme 500 mg and Flavorzyme 500L are easy to dissolve in water.

Flavorzyme activity varies with pH and temperature as shown in FIGS. 4 and 5.

Flavorzyme can be deactivated in 10 minutes at temperatures above 75 ° C. (167 ° F.). However, inactivation is quite substrate dependent (substrate concentration, pH, etc.).

Preferably the composition of the present invention is in the form of a topical application medicament or cosmetic preparation such as solution, cream, ointment, ointment stick, gel, powder or adhesive plaster. Proteolytic enzymes are mineral oils, water-based creams or lotions, oil-based creams or lotions, and other known peelings that allow the resulting mixture to maintain neutral or alkaline pH because the enzyme is inert in an excessively basic or acidic environment. It is supported on a pharmaceutically or cosmetically acceptable carrier such as a substance.

Such topical compositions may contain other active pharmaceutical ingredients such as fungicides and antibiotics, as well as conventional adjuvants such as perfumes and other cosmetic additives and excipients commonly used in cosmetic and skin products.

Compositions containing from about 1% to about 15% proteolytic enzymes in a pharmaceutically active carrier are effective at softening, thinning and removing the mammal's keratinous tissue. Pharmaceutically acceptable carriers are present in amounts of about 85% to about 99%. In a preferred embodiment, the composition contains from about 2.4% to about 10% proteolytic enzyme, depending on the application. The remainder of the composition comprises a pharmaceutically acceptable carrier in an amount of about 90% to about 97.5%.

The composition may contain, but is not limited to, other additives or excipients such as antibiotics, fungicides, anesthetics, wetting agents, flavorings, dyes and the like.

The following examples of the proteazel containing compositions described above are preferred embodiments of the invention:

Formulation # 1: A composition comprising 5% protease and 95% mineral oil, such as Alcalase®, consists of 90% enzyme component and 10% Flavorzyme ^™ . This formulation is suitable for application anywhere in the human body except face and vulva.

Formulation # 2: A composition comprising 2.4% protease and 97.5% mineral oil, such as Alcalase®, consists of 90% enzyme component and 10% Flavorzyme ^™ . This formulation is suitable for the face, especially for sensitive areas such as eyes, nose and mouth.

Formulation # 3: Composition comprising 10% proteinase enzyme and 90% mineral oil, including Alcalase® and Flavorzyme ^™ , for use in mammalian skin under the control of a veterinarian.

Formulation # 4: A composition comprising 5% proteolytic enzymes and 95% mineral oil, such as subtilisin Carlsberg, consists of 100% enzyme components. This formulation is suitable for application similar to formulation # 1.

The protease-containing composition is applied every 12 hours to calluses, nails, keratin or dry cracked skin and covered with a dressing if necessary. During application, the part is cleaned by a swelling action with a warm, wet washcloth and soap. In the case of plantar calluses, such administration is reduced to three times per week if the callus no longer causes discomfort, since this is to some extent important for bone protection. Likewise, if the keratinous condition is exfoliated, the use of ointment periodically can prevent remodeling or recurrence.

In a preferred embodiment, the present invention is disposed on a carrier such as dressing of an adhesive tape, plaster or the like, but is not limited thereto.

In the most preferred embodiment, the dressing, such as the adhesive band, is dipped by the protease containing composition according to the invention. Preferably, the dressing comprises an adhesive band by an absorbent pad, which pad is immersed by a protease such as mineral oil and a pharmaceutically or cosmetically acceptable carrier. The composition-immersion dressings are applied onto the hyperkeratinous tissue to be treated.

Experiment

Experiment 1: On day 21, a placebo-controlled study of plantar callus using Formulation # 1 rapidly improved gait comfort for all volunteers within 14 days, as shown in the photographs of FIGS. 6 and 7. Independent clinical investigators reported less than 50% callus compared to values obtained from available callus lotions, creams and ointments.

Experiment 2: Formulation # 1 was applied to the heel of a 81 year old old man with severe heel dryness, including cracks and peeling skin, and all hyperkeratomic tissues were controlled within 10 days. Illustrated in FIGS. 8 and 9 photos.

Experiment 3: Using Formulation # 1, a 50 year old male applied daily for two weeks to remove cracks in his heel. The crack-free heel was maintained by applying two or three times weekly. Heel cracks were similarly removed in young men.

Experiment 4: A 45-year-old woman applied Formulation # 1 to her thick, flaky soles that were cracked and dry. With regular use, her soles became soft and smooth.

Experiment 5: Formulation # 1 was used by a middle-aged woman to exfoliate from her instep by applying ointment twice a day for 7 days. Sore callus was also reduced.

Experiment 6: A 57-year-old woman used Formulation # 1 to remove hand callus from her horticulture. Within a day, the callus in the palms was reduced by 50%. Within 3 days, callus areas appeared normal.

Experiment 7: A middle-aged woman applied Formulation # 1 to the nails on her toes. In 48 hours, enough softening was noticeable and the discomfort of the shoes disappeared. The nail surface layer swelled mechanically and peeled off after 4 days.

Experiment 8: A formulation 1 according to the invention was used at night to remove or alleviate the appearance of age-based lines and to improve skin softness.

Experiment 9: A 58-year-old woman uses Formulation # 1 according to the invention once a week to remove wrinkles on her lips and to remove dead skin sepol in the dry areas of the body, including under the legs and hands. It was.

Experiment 10: A middle-aged woman found that the ointment of Formulation # 1 was applied to the eczema area of her foot, significantly improving the physical signs of condition and itching.

Experiment 11: An elderly woman who had severe eczema on her hands and had to wear gloves for 24 hours a day applied Formulation # 1 according to the present invention to her hands for several weeks in the morning and evening. Her hands were improved enough to show her hands to others without wearing gloves for the first time in 15 years.

Experiment 12: A female in her 30s applied Formula # 1 according to the invention every night to the cuticle of the nail. After two weeks the dry, white layer was peeled off and the new edges were smoothed to give a uniform appearance.

The present invention has been described with reference to specific embodiments, which are for illustrative purposes only. Various modifications according to the present invention will be appreciated by those skilled in the art, which should fall within the spirit and scope of the present invention. All patents and documents cited herein are hereby incorporated by reference.

Claims (32)

A composition for removing, softening or thinning the keratinous tissue of a mammal comprising at least one proteolytic enzyme for food and a pharmaceutically or cosmetically acceptable carrier. The composition of claim 1, wherein said food proteinase comprises Subtilisin Calsberg . The composition of claim 1, wherein said food proteinase comprises Flavorzyme ^™ . The composition of claim 1, wherein said food proteinase comprises Subtilisin Calsberg and Flavorzyme ^™ . The composition of claim 1, wherein the pharmaceutically or cosmetically acceptable carrier comprises a water based carrier. The composition of claim 1, wherein the pharmaceutically or cosmetically acceptable carrier comprises an oil based carrier. The composition of claim 1 further comprising a fragrance. The composition of claim 1 further comprising an antibiotic. The composition of claim 1 further comprising an antifungal agent. The composition of claim 1 further comprising an anesthetic. The composition of claim 1, wherein the pharmaceutically or cosmetically acceptable carrier comprises mineral oil. The composition of claim 1 comprising about 1% to about 15% proteolytic enzyme and about 85% to about 99% pharmaceutically or cosmetically acceptable carrier. The composition of claim 1 comprising about 5% proteolytic enzyme and about 95% mineral oil. The composition of claim 13, wherein said proteolytic enzyme comprises about 90% Subtilisin Calsberg and about 10% Flavourzyme ^™ . The composition of claim 1 comprising about 2.4% proteolytic enzyme and about 97.5% mineral oil. The composition of claim 13, wherein said proteolytic enzyme comprises about 90% Subtilisin Calsberg and about 10% Flavourzyme ^™ . The composition of claim 1 comprising about 10% proteolytic enzyme and about 90% mineral oil. The composition of claim 15, wherein said proteolytic enzyme comprises Subtilisin Calsberg and Flavorzyme ^™ . Obtaining a composition comprising at least one food proteolytic enzyme selected from the group comprising Subtilisin Calsberg and Flavorzyme ^™ and a pharmaceutically or cosmetically acceptable carrier, and Applying the composition to the keratinous tissue of a mammal; Method of treating hyperkeratin tissue in mammals. 18. The method of claim 17 comprising applying said composition to calluses. 18. The method of claim 17 comprising applying said composition to a nail. The method of claim 17 comprising applying the composition to the follicular keratin. 18. The method of claim 17, comprising applying the composition to dry or cracked skin. 18. The method of claim 17, further comprising dressing up and covering the tissue to which the composition has been applied. A substrate, at least one food proteinase and a pharmaceutically or cosmetically acceptable carrier, wherein said proteinase and carrier are disposed on said substrate to treat a mammal's keratinous tissue. Dressing for A method of treating a keratinous tissue of a mammal comprising applying to said mammalian keratinous tissue a composition comprising at least one food proteinase and a pharmaceutically or cosmetically acceptable carrier. A method of treating eczema in a mammal comprising applying to a mammalian eczema a composition comprising at least one food proteinase and a pharmaceutically or cosmetically acceptable carrier. A method of treating a mammalian epidermis comprising applying to a mammalian epidermis a composition comprising at least one food proteinase and a pharmaceutically or cosmetically acceptable carrier. Treating facial wrinkles and lines in mammalian skin comprising applying to the facial wrinkles and lines in mammalian skin a composition comprising at least one food proteinase and a pharmaceutically or cosmetically acceptable carrier; How to prevent. 27. The method of claim 26, further comprising dressing up and covering the tissue to which the composition has been applied. 27. The method of claim 26, wherein said food proteinase is selected from the group comprising Subtilisin Calsberg and Flavorzyme ^™ . 27. The method of claim 26, wherein the composition comprises about 2.4% to about 10% proteolytic enzyme and about 90% to about 97.6% carrier.