KR20060091829A - A pharmaceutical composition comprising retinoid for prevention and treatment of vitiligo - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating vitiligo comprising retinoid as an active ingredient. Specifically, the retinoid reduces the death of human keratinocytes and proliferates the number of cells, thereby causing melanocyte death or necrosis. It relates to a pharmaceutical composition for preventing and treating vitiligo prepared to show activity against vitiligo. The pharmaceutical composition comprising the retinoid of the present invention as an active ingredient not only prevents the side effects of skin atrophy induced by corticosteroids, which are conventional treatments for vitiligo, but also suppresses the death of melanocytes, thereby showing pigmentation effect. It can be usefully used for prevention and treatment.

ATRA, SCF, TUNEL assay, activated caspase, Ki67, MTT assay

Description

Pharmaceutical composition comprising retinoid for prevention and treatment of vitiligo

Right hand as the test groups A of Figure 1, treatment a pharmaceutical composition comprising a retinoid of the present invention as an active ingredient and left hand was treated as the control group, placebo of the vehicle (V) subjected to a placebo-controlled trial comparing the results It is a photograph,

FIG line graph showing the number of patients from time to time that the improved results by B is a pharmaceutical composition of the present invention shown in the first,

A top of FIG. 2 is a photograph showing Western blot comparing the expression level of stem cell factor (SCF) in the group treated with the retinoid of the present invention to the beta actin expression, and the bottom is unresponsive. SCF expression in the group is a photograph shown by Western blot compared with the expression level of beta actin,

Normal: Normal skin of patient

Vehicle: Discolored epidermis of vehicle treated patients

ATRA: depigmented epidermis in retinoid treated patients

Actin: A protein used as a standard to compare expression levels.

Figure 2B shows the relative ratio of SCF expression of the decolorized epidermis treated with vehicle and retinoid treated by vehicle (V) and retinoid (A) divided by SCF expression of normal epidermis. Bar graph,

L: discolored epidermis N: normal epidermis

FIG. 3 shows the normal and epidermal cells treated with retinoids and vehicle, respectively, and stained with TUNEL (green) and Ki67 (red) to identify apoptotic and proliferating cells. Fluorescence microscopy of the unstained group of epidermis

4 is a diagram showing the expression of these proteins using antibodies capable of detecting activated caspases 8, 9 and 3, which are proteins related to apoptosis in the epidermis of patients with improved and unresponsive groups. The three lanes of were performed in patients in the improved group and the three lanes on the right were in patients in the no response group.

N: Normal epidermis V: Vehicle decolorized epidermis A: Retinoid decolorized epidermis

5 is a bar graph showing that the number of melanocytes (optical density) does not increase when the retinoid concentration is increased to 0, 1, 100 nM in the MTT assay,

A in Fig. 6 in the MTT assay is a bar graph illustrating that the increase in the number of keratinocytes when high concentrations of retinoid as described above,

B of FIG. 6 is an illustration showing that the concentration of a retinoid increases the SCF protein expression in keratinocytes when the increase as described above.

The present invention relates to a pharmaceutical composition for the prevention and treatment of vitiligo, which comprises retinoid as an active ingredient. Specifically, the retinoid reduces the death of human keratinocytes, thereby proliferating the number of cells, resulting from the death or necrosis of melanocytes. It relates to a pharmaceutical composition for preventing and treating vitiligo prepared to show activity against vitiligo.

Vitiligo is an acquired depigmentation disease in which white patches of various sizes and shapes appear on the skin due to the death or necrosis of melanocytes. It is a relatively common disease in about 1% of the world's population, with no differences in race or location. The incidence is most common at 10-30 years, 95% of patients develop before 40 years of age, and 30% of patients have a family history.

Clinical manifestations of vitiligo include round or irregular white spots of varying size, and the degree of discoloration is not clear at first, but it is often clearer over time. It may be darker and more pronounced than. In rare cases, itching may occur. White spots may appear on areas with hairs, especially on hair and eyebrows. Vitiligo can occur anywhere on the skin, but is especially common in areas of prominent bones such as fingers, toes, knees, and elbows, around the mouth, around the nose, around the eyes, underarms, and inside the wrist. It can also come on the mucous membranes, such as the lips and genitals, and occurs particularly well on wounded areas. The distribution of vitiligo is symmetrical or neural. Vitiligo may be accompanied by abnormal pigmentation of the iris and retina of the eye, in addition to the appearance of white spots on the skin. And autoimmune diseases such as lupus erythematosus.

The cause of vitiligo has not been elucidated yet, but many theories have been suggested, including autoimmune theory, neuronal fluidism, and melanocyte self-destruction. Autoimmune theories suggest that expression of autoantibodies against melanocyte antigens results in the destruction or dysfunction of melanocytes or the destruction of melanocytes in lymphokines secreted by cytotoxic lymphocytes or activated lymphocytes. Neuronal fluid is a disorder of catecholamine biosynthesis and an increase in monoamine oxidase, which leads to stress-related hydrogen peroxide, which causes melanocytes to be destroyed, and vitiligo may occur along the ganglion or after nerve damage or stress. The melanin self-destruction theory suggests that phenol complexes, which are intermediate metabolites or final metabolites, generated during melanin formation accumulate in melanocytes and destroy cells. In addition, various factors such as inherent cellular defects, genetic factors, apoptosis, and calcium metabolism abnormality have been suggested.

Many studies of vitiligo have focused on melanocytes. However, since melanocytes in the skin form functional structural units with neighboring keratinocytes, studies on keratinocytes in association with vitiligo are needed. Indeed, the activity of catalase, glutathione reductase (challreuter KU et al., J Invest Dermatol 97: 1081-1085, 1991) and the calcium uptake (Schallreuter KU et al., Arch Dermatol Res 280: 137-139, 1988) Functional abnormalities, saturation, and structural abnormalities resulting from the degeneration of intracellular granules into the extracellular space were found in keratinocytes of vitiligo (Moellmann G. et al, J. Invest Dermatol 79: 321-330, 1982 Bhawan J. et al., J Cutan Pathol 10: 207-212, 1983). The saturation of keratinocytes is known to be caused by damage caused by hydrogen peroxide (Maresca V. et al., J. Invest Dermatol 109; 310-313, 1997). In fact, hydrogen peroxide is known as keratinocytes. It is one of the most potent stimulants (Kucchle MK et al., Cell Death Differenciation 7; 566-573, 2000) that causes the death of.

 Currently commonly used treatment methods for vitiligo include psoralen ultraviolet therapy, local and systemic treatment of steroids, and surgical treatment. Soralene UV therapy is called photochemotherapy, and it is systemic ultraviolet irradiation after taking soralene or partial ultraviolet ray irradiation after applying soralene. Since it requires long-term treatment, there is a risk of skin aging or skin cancer caused by ultraviolet irradiation. Steroid treatment requires topical application or topical injection with triamcinolone when the lesion area is small, and systemic administration if widespread. This also requires long-term treatment and there is a risk of causing skin and systemic side effects. Surgical treatment is a method of transplanting melanocytes by surgery, which can be divided into skin graft and cell graft, and for the skin graft, full layer, middle layer and epidermal graft are used. This method is not always available and can only be used if the lesion has not spread for years.

The most commonly used method is the treatment using an external steroid, and if it is used for a long time, side effects including skin contraction appear, so a method for solving the problem is required.

 Retinoic acid has been used mainly for acne treatment because it can suppress the formation of scrim and open pores. It also reduces melanocyte migration and adhesion, inhibits growth (Fligiel SE et al., J. Cutan Pathol 19; 27-33, 1992) and reduces melanocyte dendritic processes to cloud pigment. (Ortonne JP, Br J Dermatol 127 suppl 41: 43-47, 1992) Retinoic acid may be used to suppress the side effects of corticosteroids, as reports have recently been reported to reduce skin contraction caused by corticosteroids. It is expected to be.

Accordingly, the present inventors prepare a pharmaceutical composition comprising the retinoid as an active ingredient, and the composition prevents skin contraction caused by corticosteroids, as well as proliferates human keratinocytes and inhibits apoptosis, thereby SCF and melanocytes. By increasing the amount of the factor for growth, the proliferation and staining of melanocytes confirmed that it is effective in treating vitiligo and completed the present invention.

The present invention provides a pharmaceutical composition for the prevention and treatment of vitiligo, which has retinoids as an active ingredient and prevents the side effects of skin contraction induced by corticosteroids, as well as proliferates melanocytes and shows a pigmentation effect.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of vitiligo, using retinoid as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for the prevention and treatment of vitiligo comprising retinoids as an active ingredient.

The retinoid, which is an active ingredient of the composition of the present invention, may be selected from the group consisting of retinol, retinal, retinoic acid, tazarotene and adapalene, preferably using retinoic acid. . Retinoic acid includes tretinoin, isotretinoin and etretinate, and in the preferred embodiment of the present invention, tretinoin was used.

In order to confirm whether the pharmaceutical composition including the retinoid of the present invention has an improvement effect on the lesion area of vitiligo, 12 months of administration of tretinoin, a type of retinoid, was performed in real patients. In particular, 30.6% of patients showed improvement within 6 months.

In addition, the present inventors confirmed whether the composition containing the retinoid of the present invention affects the stem cell factor (SCF) value of the epidermal cells of keratinocytes. To this end, samples of normal epidermis and bleached epidermis treated with tretinoin and vehicle, respectively, were prepared, and expression values were measured using antibodies of SCF. As a result, the SCF level was lower than the normal cuticle in both the tretinoin treated and vehicle treated decolorized epithelium, and compared with the depigmented epidermis, the tretinoin treated epidermis was higher than the vehicle treated epidermis. Similar in the unresponsive group.

In addition, the present inventors confirmed whether the retinoid of the present invention affects the expression level of activated caspases 3, 8, and 9, which are proteins related to apoptosis and apoptosis. To this end, sections of the normalized epidermis and each of the depigmented epidermis treated with tretinoin and vehicle were prepared and subjected to TUNEL assay to determine the apoptosis and the expression level of activated caspase was examined by the same method as the SCF. As a result, the number of TUNEL positive cells in the discolored epidermis was increased or similar compared to normal epidermis. Compared to the depigmented epidermis, the tretinoin-treated epidermis had fewer TUNEL-positive cells than the vehicle-treated epidermis in the improved group, and the tretinoin-treated epidermis had similar TUNEL-positive cells than the vehicle-treated epidermis. Activated caspases 3, 8, and 9 showed increased or similar expression of the depigmented epidermis compared to normal epidermis, and compared with the depigmented epidermis, tretinoin treated group showed lower expression level than vehicle treated group. In the unresponsive group, the expression level was similar.

In addition, the inventors confirmed whether the retinoids of the present invention affect the proliferation of cells. To this end, Ki67 detection was performed to identify cells that proliferated by making sections of each of the normalized epidermis, the depigmented epidermis treated with tretinoin and the vehicle. As a result, the number of Ki67-positive cells was less than that of normal epidermis in vehicle-treated bleached epidermis, but the tretinoin-treated epidermis was more or similar to that of normal epidermis. Compared with the depigmented epidermis, the tretinoin-treated side was similar or slightly more Ki67-positive than the vehicle-treated side. These results were found in both improved and unresponsive groups with varying degrees.

 In addition, the present inventors confirmed whether the retinoid of the present invention affects the proliferation of these cells when cultured keratinocytes and melanocytes of normal people. For this purpose, we used an MTT assay kit to detect cell growth by spectroscopic method and examined whether the light density increased in keratinocytes and melanocytes by increasing the concentration of tretinoin. As a result, as the concentration of tretinoin increased, light density in keratinocytes increased but not melanocytes. In other words, the number of keratinocytes increased but the number of melanocytes did not increase as the concentration of tretinoin increased.

In addition, the present inventors confirmed whether the retinoid of the present invention affects the SCF expression level of keratinocytes of normal people cultured. For this purpose, expression levels were measured using SCF antibodies. As a result, the level of SCF increased with increasing concentration of tretinoin.

Therefore, the present invention can be useful for the prophylaxis and treatment of vitiligo by proliferating and staining melanocytes by proliferating human keratinocytes and inhibiting apoptosis, thereby increasing the amount of factors for SCF and melanocyte growth. have.

The pharmaceutical composition for preventing and treating vitiligo of the present invention may further contain at least one active ingredient having the same or similar function in addition to the retinoid. The pharmaceutical composition may be prepared by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredient for administration. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, if necessary, as antioxidants, buffers And other conventional additives such as bacteriostatic agents can be added. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to formulate injectable formulations such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA.

The pharmaceutical composition of the present invention is preferably parenterally administered (e.g., applied to the intravenous, subcutaneous, intraperitoneal, topical or visual route), more preferably topical administration. For topical administration, for the treatment of the skin and mucous membranes, it is possible in the form of paste ointments, creams, milks, powders, penetration pads, solutions, gels, sprays, lotions or suspensions. They may also be in the form of microparticles or microparticles, or fatty or polymeric vesicles or polymeric additives and hydrogels capable of controlled release. Compositions of topical administration may be anhydrous or aqueous, depending on the clinical prescription.

Dosage varies depending on the weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion and severity of the patient. The pharmaceutical composition of the present invention comprises 0.001 to 1% by weight of retinoids, and more preferably administered once to several times a day.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the present invention more easily, but the content of the present invention is not limited to the following examples.

Experimental Example 1 A Placebo-Controlled Experiment on the Treatment of Tretinoin in Patients with Vitiligo

A placebo-controlled paired comparison left-right study was performed in which a specific experiment was performed in pairs with a control group treated with a placebo (vehicle) and a control group treated with tretinoin. Sixty-two patients with vitiligo were treated with tretinoin on one hand, arms, legs, torso, abdomen, and face, and on the other with a vehicle manufactured by the same company (Stiefel Labs. Ltd., Singapore). After applying tretinoin or vehicle, apply topical corticoid once or twice a day, and then, at the depigmentation site treated with tretinoin and the depigmentation site treated with vehicle, The extent to which the pigment was reformed as a whole was compared. Comparative photographs were taken every two weeks for three to six months. Two dermatologists judged the results, and the experiment was discontinued if the symptoms in one of the experimental groups improved further.

As a result, 27 out of 62 patients showed better repimentation on the tretinoin treated side than on the vehicle treated side (FIG. 1A) . The group showing improved results with tretinoin was named the improved group, and the group showing no improved results with tretinoin was designated as an unresponsive group. In the group that improved after tretinoin treatment, four (6.5%) within one month, nine (14.5%) within three months, 13 (21%) within four months, 17 (27.4%) within five months, and six months In 19 patients (30.6%), improvement in response was evident (monthly cumulative). Although three patients (4.8%) showed a difference in clinical results about a year later, the difference in effect was rapid in most patients (FIG. 1B) .

Experimental Example 2 Western Blot Analysis of SCF

Normally colored epidermis and depigmented epidermis treated with tretinoin and vehicle respectively were frozen in liquid nitrogen and the frozen tissues were ground to powder. Epidermal specimens were treated with 50 mM Tris-base (pH 7.4), 150 mM NaCl, 10 mM EDTA, 0.1% Tween-20 and protease inhibitors (0.1 mM phenylmethylsulfonyl- fluoride, 5 μg / ml aprotinin, and 5 μg / ml leupeptin). It was homogenized in a buffer solution containing and iced. The homogenate was centrifuged at 12,000 g, 4 ° C. for 30 minutes, and the supernatant was collected. The concentration of protein in the supernatant was measured using a DC protein analysis kit (Bio-Red Laboratories, Inc., Herculus, CA). The same amount of protein, 20 μg each, was separated using 7% SDS-PAGE and transferred to nitrocellulose membrane. 5% non-fat dry milk (NFDM) dried in TTBS (Tris-buffered saline) containing 10 mM Tris (pH 7.6), 150 mM NaCl and 0.1% Tween-20 W / V) The membrane was immersed in the added blocking solution and placed at room temperature for 1 hour. The membrane was immersed in an anti-Human SCF (mouse monoclonal; Santa cruz, CA) antibody diluted in blocking solution at a ratio of 1 / 1,000 and reacted overnight at 4 ° C. After rinsing with TTBS, it was immersed in anti-Rabbit or anti-mouse antibody (Jackson Laboratories, West Grove, PA) with horseradish peroxidase diluted 1 / 1,000 in blocking solution. Put on for 1 hour. This was washed with TTBS for 30 minutes and then treated with chemiluminescent solution (ECL Kit; Amersham Life Science, Buckinghamshire, England) for 1 minute and exposed to X-ray film (Hyperfilm; Amersham Life Science, Buckinghamshire, England). To determine the amount of protein, it was re-reacted with an antibody capable of detecting beta actin. Using anti-actin (mouse monoclonal) diluted with 1 / 5,000 in blocking solution, the previous antibodies attached to the protein on the membrane surface were removed using a stripping buffer (0.1 M Glycine, pH 2.5). Sigma, St. Louis, MO) soaked in antibodies and placed at room temperature for 1 hour. Subsequently, secondary antibody treatment and subsequent processes were performed by the same experimental method as described above. The film was developed immediately to analyze the concentration of protein bands.

As a result, SCF levels were lower in the depigmented epidermis than in normal epidermis (FIG. 2A), and SCF levels at tretinoin treated sites than vehicle treated sites in 7 patients in the improved group of 13 patients who measured SCF. Is significantly higher (Fig. 2A Top and FIG. 2B), there were no significant differences in SCF values in 6 patients in the no-response group (bottom of FIG . 2A and FIG .

Experimental Example 3 Apoptosis and Cell Proliferation Confirmation Experiment

1) TUNEL assay

The epidermal specimens immobilized with 4% paraformaldehyde were dehydrated and then placed in paraffin wax, cut into 3 μm thick sections, deparaffinized with xylene, and subsequently immersed in a series of alcohol solutions for rehydration. Normal and epidermal apoptosis cells treated with tretinoin and vehicle were examined by TUNEL method (dUTP-digoxigenin nick end-labeling; In situ cell death detection kit, Roche, Mannheim, Germany). Sections were washed with PBS and then pretreated with proteinase K (Proteinase K; Invitrogen, Carlsbad, Calif.) For 15-30 minutes. It was washed again with PBS, and reacted for 1 hour at 37 ℃ in the TUNEL reaction mixture contained in the kit. After washing with distilled water, nuclei were counterstained with Hoechst 33258 (Sigma, St. Louis, Mo.) and observed under a fluorescence microscope (Nikon Eclipse TE300, Japan). For quantitative analysis of cells in which apoptosis occurs, the normal epidermis, tretinoin, and vehicle-treated depigmented epidermis are randomly selected and photographed at four sites, and then the number of apoptotic cells and the total number of cells are examined. It was. The number of apoptotic cells was expressed in% relative to the total number of cells (apoptotic cells / total cells).

As a result, TUNEL-positive cells were found in the lower and upper granule layers of the epidermis and the number of cells showed distinct differences between the two groups. In the improved group, the number of TUNEL-positive cells in the tretinoin-treated bleached epidermis was lower than that of the vehicle-treated bleached epidermis ( top of FIG. 3 ) , whereas in the non-reactive group, the tretinoin-treated bleached epidermis was less than the vehicle-treated bleached epidermis The number of TUNEL positive cells was similar ( bottom of FIG. 3 ) .

2) Immunohistochemical staining for Ki67 and double staining using TUNEL method

For immunofluorescence staining of Ki67, bleached epidermis treated with tretinoin and vehicle together with normal epidermis was cut into 3 μm thick sections. After deparaffinization and rehydration, the sections were soaked in citric acid solution (100 mM citrate, pH 6.0) for 1 minute and then washed with Tris buffer. Sections were soaked in a solution containing 10% fetal Bovine Serum and placed at room temperature for 2 hours. Anti-Ki67 (mouse monoclonal; Zymed, South San Francisco, CA) antibody diluted 1: 200 was then reacted overnight at room temperature. The sections were washed with PBS and then reacted with biotin-bound anti-mouse IgG (Jackson ImmunoResearch Laboratories, West Grove, PA) antibody for 2 hours. After the reaction, the cells were washed with PBS, and the fragments were reacted with streptavidin with Alexa Fluor 594 conjugate (Molecular Probes, Eugene, OR). Sections were double-stained again by the same TUNEL method as described above, and the results were confirmed using a fluorescence microscope (Nikon Eclipse TE300, Japan).

As a result, the Ki67-positive cells were scattered in the lower part of the epidermis and the number of Ki67-positive cells increased with the difference between the tretinoin-treated side and the vehicle-treated side. These results showed no significant difference between the improved and unresponsive groups. In the group that improved with double staining with TUNEL, the tretinoin-treated bleached epidermis had a significantly smaller number of TUNEL-positive cells and more Ki67-positive cells than the vehicle-treated bleached epidermis ( top of FIG. 3 ) . On the other hand, in the non-responsive group, the discolored epidermis treated with tretinoin was similar to the discolored epidermis treated with vehicle and the number of cells positive for Ki67 was also slightly higher or similar ( bottom of Figure 3 ) . In other words, in the group affected by tretinoin, tretinoin significantly reduced cell death and slightly increased cell proliferation, thereby increasing the number of cells as a whole.

Experimental Example 4 Western Blotting Assay for Activated Caspase

Activated caspase 3 (rabbit polyclonal antibody; Pharmingen, San Diego, CA), caspase 8 (rabbit polyclonal antibody; Pharmingen, San Diego, CA) and caspase 9 (rabbit polyclonal antibody; Pharmingen, San Diego, CA) Experiments were performed in the same manner as in the Western blot of SCF described above, except for each primary antibody.

As a result, the expression of these proteins in vehicle-treated or tretinoin-treated bleached epidermis increased compared to normal epidermis (FIG. 4) . In the improved group, tretinoin treatment showed lower levels of activating caspase 3, 8 and 9 proteins (apoptosis related proteins) than vehicle treated. On the other hand, in the unresponsive group, the expression level was similar (FIG. 4) .

Experimental Example 5 MTT Analysis

<5-1> Melanocyte Culture of Normal Persons

A sample of skin from a normal person was treated with dispase (# 295 825, Roche, Mannheim, Germany) for 1 hour to separate the epidermis from the dermis. This epidermal layer was trypsinized for 10 minutes to form a single epidermal cell solution. After centrifugation, the cells were cultured in MCDB 153 medium containing vitamin E, 12-O-tetradecanoylphorbol 13-acetate, pituitary gland extract, insulin, transferrin, selenium and basal fibroblast growth factor. Once the cells were attached to the bottom of the culture flask, SCF was added. When the cells proliferated, they were separated from the flasks and inoculated into fresh culture flasks. Before detaching melanocytes, XB-2 cells were prepared as follows. XB-2 cells were grown for 4 hours with 10 μg mitomycin C without 3T3 feeder cells and treated with trypsin to make cell suspensions of each individual state. Cells were then seeded in fresh culture flasks (5 × 10 ⁴ cell count / ml). Melanocytes passaged 3 to 10 times were used for the experiment.

<5-2> Culture of keratinocytes of normal individuals

Single epidermal cell solution was prepared and centrifuged in the same manner as in melanocyte culture.The cells were then treated with KGM medium containing bovine pituitary extract, human epidermal growth factor, insulin and hydrocortisone (# CC-3101; BioWhittaker, Walkersville). , MD). When the cells proliferated, they were separated from the flasks and inoculated into other culture flasks.

<5-3> MTT assay

MTT assay kit (# 4890-25-01, R & D, Minneapolis, MN) was used to investigate the effects of tretinoin on the proliferation of melanocytes. The concentrations of tretinoin applied to the assay are 0, 1 and 100 nM. The effect on the proliferation of keratinocytes was also investigated using the MTT assay kit. The concentrations of tretinoin applied to the assay are 0, 1 and 100 nM.

As a result, melanocytes did not proliferate or decrease at tretinoin at a concentration of 0-100 nM (FIG. 5) , while keratinocytes proliferated (FIG. 6A) . Tretinoin at a concentration of 0-100 nM also increased the levels of SCF produced in keratinocytes (FIG. 6B) .

Examples of preparations for the compositions of the present invention are illustrated below.

Formulation Formulation for Topical Administration

Ointment

0.020 g of retinoic acid

81.700 g of isopropyl myristate

Mineral oil fluid 9.1 g

9.180 g of silica

2. Cream

0.100 g of retinoic acid

Lanolin emulsifying alcohol, wax 39.900 g

0.075 g of methyl para-hydroxybenzoate

0.075 g of propyl para-hydroxybenzoate

Sterile demineralized water balance 100 g

3. Lotion

0.100 g of retinoic acid

69.900 g of polyethylene glycol (PEG 400)

30.000 g of 95% ethanol

As described above, the retinoid of the present invention not only prevents skin contraction by topical corticosteroids, but also proliferates and inhibits apoptosis of keratinocytes of humans, thereby reducing SCF and related factors for melanocyte growth. By increasing the amount, melanocytes can be proliferated and stained, which can be useful for the prevention and treatment of vitiligo.

Claims (4)

Pharmaceutical composition for the prevention and treatment of vitiligo comprising retinoids as an active ingredient. The method of claim 1, wherein the retinoid is a group consisting of retinol, retinal, retinoic acid, tazarotene and adapalene pharmaceutical composition for preventing and treating vitiligo. The pharmaceutical composition for preventing and treating vitiligo according to claim 2, wherein the retinoic acid is tretinoin, isotretinoin, and etretinate. The pharmaceutical composition for preventing and treating vitiligo according to claim 1, wherein the composition is a topical formulation in the form of a paste ointment, cream, milk, powder, penetration pad, solution, gel, spray, lotion or suspension.

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