TW201632185A - Skin whitening composition prepared with imatininb and application thereof - Google Patents

Abstract

The present invention relates to a skin whitening composition prepared with imatinib and the application thereof. The composition comprises imatinib, alcohols, nut oils, polysaccharide complex, esters, and pharmaceutically acceptable carrier, excipient, thinner, and adjuvant. The composition uses the approach of reducing melanin content in cells through inhibiting tyrosinase activity to achieve the efficacy of skin whitening.

Description

Preparation of skin whitening composition using imatinib and use thereof

The present invention relates to an imatinib composition and its use, and in particular to a composition for skin whitening using imatinib and uses thereof.

In general, skin color is determined by several physiological factors, one of which is the amount of melanin present in the skin. If the melanin content is higher, the skin tone will be darker. Therefore, the research and development direction of the whitening field in the past few decades has focused on how to reduce the content of melanin in the skin, in order to achieve skin whitening effect. Tyrase is an important enzyme in melanin biosynthesis, so if the activity of tyrosinase is inhibited or reduced, the amount of melanin synthesized can be reduced. There are a variety of enzymes in the cell as well as metabolic pathways that can be used to minimize oxidative damage to cells. Ultraviolet radiation can cause protein oxidation, damage to DNA, and the formation of reactive oxygen molecules in the skin, resulting in age-related diseases or even melanin production.

Melanin is produced from melanocytes, and the main function of melanin is to protect the skin from ultraviolet radiation. However, the production of melanin excess and its accumulation in the skin can cause a variety of skin lesions such as dark spots, age spots, freckles and other symptoms associated with hyperpigmentation. Tyrosinase is a key enzyme in the process of melanin production that affects the rate-determining step. Among them, this step is to convert L-tyrosine to L-DOPA, and levodopa can be further oxidized to O-quinone. Therefore, tyrosinase is the primary target for screening inhibitors that inhibit melanin production.

In recent years, many inhibitors of melanin production have been widely used in skin care products for preventing pigmentation. Surprisingly, research reports indicate that hydrogen peroxide and other reactive oxygen species (ROS) are produced during melanin production, and that these reactive oxygen molecules cause strong oxidative stress in melanocytes. ). Reactive oxygen molecules (ROS) such as singlet oxygen, molecular oxygen, hydrogen peroxide (H ₂ O ₂ ), etc., all initiate oxidation reactions and generate free radical molecules. Free radicals are atoms, molecules or ions that have independent and unpaired electrons. For example, superoxide anion, hydroxyl radical, and peroxyl radical are three common free radicals with short period properties and chemical activity. On the other hand, active oxygen molecule scavengers (ROS scavengers) and inhibitors of reactive oxygen species production reactions inhibit UV-induced melanin production, which represents an important role for reactive oxygen species in the regulation of melanin production. Therefore, many commercially available whitening products contain an antioxidant such as vitamin C and its derivative, or reduced glutathione (GSH) to inhibit the melanin production reaction.

All scientific and technical terms used in this specification, unless otherwise defined, are defined by those of ordinary skill in the art.

The primary object of the present invention is to provide a use of imatinib for the preparation of a composition for skin whitening.

Wherein the composition comprises: imatinib, an alcohol, a nut oil, a complex polysaccharide, an ester, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, and the like.

Wherein the composition reduces intracellular melanin content by inhibiting tyrosinase activity, and the composition comprises 0.1 to 2% of imatinib.

Wherein the alcohol in the composition comprises at least one of the following: C16-18 alcohol, butylene glycol, pentanediol, octanediol, glycerol, 16 alcohol, 18 alcohol, 22 alcohol, propylene glycol.

Wherein the esters in the composition comprise at least one of the following: olive oil cetyl alcohol ester (OLIVEM 1000), glyceryl monostearate (GSM), kapok ester, isopropyl myristate (IPM) ), isopropyl palmitate (IPP), triglyceride.

Wherein the complex polysaccharide in the composition is xanthan gum, and the nut oil comprises at least one of the following: argan oil, Hawaiian stone oil, avocado oil, wheat germ oil, olive oil.

Moreover, the composition is used in the form of whitening lotion, lotion, essence, sunscreen lotion, barrier cream, and mask.

A second object of the invention is to provide a skin lightening composition.

Wherein the composition comprises: imatinib, an alcohol, a nut oil, a complex polysaccharide, an ester, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, and the like.

Wherein the composition reduces intracellular melanin content by inhibiting tyrosinase activity, and the composition comprises 0.1 to 2% of imatinib.

Wherein the alcohol in the composition comprises at least one of the following: C16-18 alcohol, butylene glycol, pentanediol, octanediol, glycerol, 16 alcohol, 18 alcohol, 22 alcohol, propylene glycol.

Wherein the esters in the composition comprise at least one of the following: olive oil cetyl alcohol ester (OLIVEM 1000), glyceryl monostearate (GSM), kapok ester, isopropyl myristate (IPM) ), isopropyl palmitate (IPP), triglyceride.

Wherein the complex polysaccharide in the composition is xanthan gum, and the nut oil comprises at least one of the following: argan oil, Hawaiian stone oil, avocado oil, wheat germ oil, olive oil.

Moreover, the composition is used in the form of whitening lotion, lotion, essence, sunscreen lotion, barrier cream, and mask.

Figure 1 Extracellular inhibition of tyrosinase activity after L-vitamin C treatment

Figure 2 Extracellular inhibition of tyrosinase activity after treatment with whitening milk

Figure 3 The intracellular melanin content of L-vitamin C treatment

Figure 4: Melanin content in cells after treatment with whitening milk

Figure 5 Intracellular inhibition of tyrosinase activity after treatment with L-vitamin C

Figure 6 Intracellular inhibition of tyrosinase activity after treatment with whitening milk

The main object of the present invention is to provide a use of imatinib for the preparation of a composition for skin whitening, the composition comprising: imatinib, an alcohol, a nut oil, a complex polysaccharide, an ester, and A pharmaceutically acceptable carrier, excipient, diluent, adjuvant, etc.; and the composition reduces the intracellular melanin content by inhibiting tyrosinase activity, thereby achieving skin whitening efficacy.

Wherein the composition has an imatinib content of 0.1 to 2%, and the composition is The alcohol is selected from at least one of the following: C16-18 alcohol, butanediol, pentanediol, octanediol, glycerol, 16 alcohol, 18 alcohol, 22 alcohol, propylene glycol; esters in the composition It is selected from at least one of the following: olive oil cetyl alcohol ester (OLIVEM 1000), glyceryl monostearate (GMS, Glycol stearate), kapok ester, isopropyl myristate (IPM), palmitic acid Isopropyl palmitate (IPP), triglyceride; the complex polysaccharide in the composition is xanthan gum; the nut oil in the composition is selected from at least one of the following: argan oil, Hawaiian stone oil, avocado Oil, wheat germ oil, olive oil.

The composition can be applied to whitened skin parts by means of whitening lotion, lotion, essence, sunscreen lotion, barrier cream and mask.

A second object of the present invention is to provide a skin whitening composition comprising: imatinib, an alcohol, a nut oil, a complex polysaccharide, an ester, and a pharmaceutically acceptable carrier, excipient, A diluent, an adjuvant, etc.; and the composition reduces the intracellular melanin content by inhibiting tyrosinase activity, thereby achieving skin whitening effect.

In the embodiment of the present invention, the whitening emulsion composition for inhibiting melanin formation is prepared by the following A agent and B agent mixture:

Agent A: Imatinib Mesylate 200mg

Agent B: OLIVEM 1000 5g, GMS 2g, 16-18 alcohol 2g, Ala Congo oil 4g, wood Cotton ester 6g, 034 4g, water 56.6g, 1,3-butanediol 5g, LIPONIC EG-1 5g, xanthan gum 0.2g, A-47 5g, BSASM 2g, pentanediol 3g, octanediol 0.2g.

The L-Vitamin C composition is used as a positive control for inhibiting the formation of melanin, and the formulated components are as follows: A agent and B agent mixture:

Agent A: L-Vitamin C 0.42g

Agent B: A-01 0.03ml, nanoHA 0.02ml, water 8.6ml, glycerol 0.3ml, 1,3-butanediol 0.4ml, phagocytosis 0.3g, symbiocell 0.3ml, PE-9010 0.05ml, octanediol 0.02 Ml

The present invention is exemplified by the following examples, but the present invention is not limited by the following examples.

Example 1 Analysis of extracellular inhibition of tyrosine tyrosine activity

Tyrosinase acts as a catalyst for the rate limiting step in melanin formation and is the most important enzyme for melanin formation. Therefore, if it can effectively inhibit tyrosinase activity, it will reduce the production of melanin and achieve the purpose of whitening. In the past, when testing the whitening effect, the mushroom tyrosine (Mushroom Tyrosinase) was the most commonly used for enzyme activity evaluation, and tyrosine (LyDosine) or levodopa (L-Dopa) was used as the substrate for the enzyme. When the enzyme catalyzes the reduction of the formation of the formed product, it can be preliminarily determined that the test sample may have whitening activity (Yeon et al ., 2008).

Referring to the method of Yeon et al ., add 40 μl of test sample (positive control group: L-Vitamin C, whitening milk) and 40 μl of tyrosinase (2Units, 1 Unit for conversion per minute under maximum activity of the enzyme) in a 96-well plate. After the micromolar (μM) substrate enzyme amount and 120 μl of 5 mM L-Dopa solution, the reaction was carried out at 37 ° C for 30 minutes, and then the absorbance at a wavelength of 490 nm was measured; and in 1% DMSO (v/v) ( Dimethylammonium) was used as a control group (Yeon et al ., 2008).

Tyrosinase inhibition rate (%) = (1 - absorbance of OD ₄₉₀ of the test sample / OD ₄₉₀ absorbance of the blank group) × 100%.

The results show that the higher the concentration of the sample used, the lower the tyrosinase activity, that is, the higher the sample concentration, the better the inhibition effect. When the concentration of L-vitamin C was 31250μg/ml, 62500μg/ml and 125000μg/ml, it had 29.05%, 45.71% and 70.60% inhibition of tyrosinase respectively (Fig. 1); and when the concentration of whitening milk was 7812.5μg/ The inhibition rates at ml, 15625 μg/ml, and 31250 μg/ml were 2.02%, 7.38%, and 13.93%, respectively (Fig. 2).

Example 2 Analysis of intracellular melanin content

The invention utilizes samples of different concentrations to detect whether the sample has an effect on the formation of melanin content in B ₁₆ F ₁₀ cells; since melanin is dissolved at _1O NaOH at 60 ° C, it has an absorbance at a wavelength of 405 nm, so It is known whether samples of different concentrations have an inhibitory effect on the production of melanin content.

Take 5×10 ⁴ cells (800 μl) of B ₁₆ F ₁₀ cells per well and 100 nM α-MSH (alpha-melanin stimulating hormone, melanin) in a 24-well plate and incubate at 37 ° C, 5% CO ₂ . After the hour, 800 μl of different concentrations of the test sample were added, and after incubation at 37 ° C, 5% CO ₂ for 24 hours, after washing twice with PBS (phosphate buffer), cells were separated using Trypsin (trypsin)-EDTA. After centrifugation at 1000 rpm for 5 minutes, 100 μl of 1 N NaOH was added and allowed to stand in a water bath at 60 ° C for 1 hour, and then allowed to stand at room temperature for cooling, and the absorbance at a wavelength of 405 nm was measured. 1% DMSO (v/v) was used as a control group.

Melanin production inhibition rate (%) = (1 - absorbance of OD ₄₀₅ of test sample / OD ₄₀₅ absorbance of blank group) × 100%.

The results showed that the higher the concentration of the sample used, the lower the melanin content in B ₁₆ F ₁₀ cells, that is, the higher the sample concentration, the better the effect of reducing melanin synthesis. When the concentration of L-vitamin C was 1.2 mg/ml, 2 mg/ml, 4 mg/ml, the melanin content was 80.54%, 79.57%, and 66.96%, respectively (Fig. 3); when the whitening milk concentration was 1.2 μg/ml, 2 μg/ml. 4μg/ml, the melanin content was 87.50%, 83.33%, and 82.64%, respectively (Fig. 4).

Example 3 Intracellular inhibition of tyrosine tyrosine activity analysis

When the tyrosinase in B ₁₆ F ₁₀ cells is collected, the principle of Freeze-Thaw Cycle is used, and the cells are frozen at -80 ° C for a period of time (30 minutes) to form ice crystals in the cells; After thawing for a period of time (30 minutes) at room temperature, the ice crystals are returned to moisture, and the cells are ruptured by the formation of ice crystals and the force of thawing, releasing the tyrosinase in the cells. The lower the value, the stronger the ability of the sample to inhibit tyrosine chymase activity.

Referring to Kim's method, 5×10 ⁴ cells (800 μl) of B16F10 cells and 100 nM α-MSH per well were placed in a 24-well plate, and cultured at 37 ° C, 5% CO ₂ for 24 hours, then 800 μl of different concentrations were added. The test sample was further cultured at 37 ° C, 5% CO ₂ for 24 hours, and then washed twice with PBS, and then the cells were detached using Trypsin-EDTA, and after centrifugation at 1000 rpm for 5 minutes, 100 μl of cells were added. Lysate (containing 50 mM PBS, 1% Triton X-100 and 0.1 mM PMSF (methyl toluene sulfonium fluoride) isopropanol solution), chilled at 80 ° C for 30 minutes, thawed at room temperature for 30 minutes (repeated 2 times), then centrifuge the cell extract at 12000 rpm for 30 minutes at 4 ° C; take 80 μl of the supernatant, then add 20 μl of L-Dopa (2 mg / ml) in a 96-well plate at 37 ° C After 10 minutes of reaction, the absorbance at a wavelength of 492 nm was measured for 1 hour (10 minutes/time). 1% DMSO (v/v) was used as a control group (Kim et al ., 2007. Antimelanogenic and antioxidant properties of Gallic acid. Biological & pharmaceutical bulletin, 30(6), 1052-1055).

Inhibition rate of intracellular tyrosinase activity (%) = (1 - absorbance of OD ₄₉₂ of test sample / OD ₄₉₂ absorbance of blank group) × 100.

The results showed that the higher the concentration of the sample used, the lower the activity of tyrosinase in B ₁₆ F ₁₀ cells, that is, the higher the sample concentration, the better the inhibitory effect. When the concentration of L-vitamin C was 1.2 mg/ml, 2 mg/ml, 4 mg/ml, the melanin content was 94.92%, 92.72%, and 86.09%, respectively (Fig. 5); when the whitening milk concentration was 1.2 μg/ml, 2 μg/ml. 4μg/ml, the melanin content was 80.54%, 79.57%, and 66.96%, respectively (Fig. 6).

In summary of the above examples, the present invention provides a pharmaceutical composition of imatinib which is a composition which can be absorbed by cells and inhibits intracellular tyrosinase activity and reduces intracellular melanogenesis. .

Based on the above effects, the present invention is a potential whitening composition for the maintenance of beauty, fully complying with the statutory invention patent requirements of novelty and progressiveness, and submitting an application according to law, and requesting the approval of the invention patent application by the bureau to encourage invention.

Claims (16)

A use of imatinib for the preparation of a composition for skin whitening, the composition comprising: imatinib, an alcohol, a nut oil, a complex polysaccharide, an ester, and a pharmaceutically acceptable carrier, Excipients, diluents, adjuvants, and the like. The use of imatinib for the preparation of a composition for skin whitening according to the first aspect of the patent application, which reduces the intracellular melanin content by inhibiting tyrosinase activity. The use of imatinib for the preparation of a composition for skin whitening according to the first aspect of the patent application, wherein the composition comprises 0.1 to 2% of imatinib. The use of imatinib for the preparation of a composition for skin whitening according to the first aspect of the patent application, wherein the alcohol comprises at least one of the following: C16-18 alcohol, butanediol, pentanediol, bisphenol Alcohol, glycerol, 16 alcohol, 18 alcohol, 22 alcohol, propylene glycol. The use of imatinib for the preparation of a composition for skin whitening according to the first aspect of the patent application, wherein the ester comprises at least one of the following: olive oil cetyl alcohol ester (OLIVEM 1000), monostearic acid Glyceryl ester (GSM), kapok ester, IPM, IPP, triglyceride. The use of imatinib for the preparation of a composition for skin whitening according to the first aspect of the patent application, wherein the composite polysaccharide is xanthan gum. The use of imatinib for the preparation of a composition for skin whitening according to the first aspect of the patent application, wherein the nut oil comprises at least one of the following: argan oil, Hawaiian stone fruit oil, avocado oil, wheat germ oil, olive oil. The use of imatinib for the preparation of a composition for skin whitening according to the first aspect of the patent application, wherein the composition is administered as a whitening lotion, a lotion, an essence, a sunscreen lotion, a barrier cream, and a mask. . A composition for skin whitening, the composition comprising: imatinib, an alcohol, a nut oil, a complex polysaccharide, an ester, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, etc. . A composition for skin whitening according to claim 9 which reduces the intracellular melanin content by inhibiting tyrosinase activity. A composition for skin whitening according to claim 9 of the invention, wherein the imatinib is contained in an amount of 0.1 to 2%. A composition for skin whitening according to claim 9 wherein the alcohol comprises at least one of the following: C16-18 alcohol, butylene glycol, pentanediol, octanediol, glycerol, 16 alcohol, 18 alcohol, 22 alcohol, propylene glycol. A composition for skin whitening according to claim 9 wherein the ester comprises at least one of the following: olive oil cetyl alcohol ester (OLIVEM 1000), glyceryl monostearate (GSM), kapok Ester, IPM, IPP, triglyceride. A composition for skin whitening according to claim 9 of the invention, wherein the composite polysaccharide is xanthan gum. A composition for skin whitening according to claim 9, wherein the nut oil comprises at least one of the following: argan oil, Hawaiian stone oil, avocado oil, wheat germ oil, olive oil. A composition for skin whitening according to claim 9 of the invention, wherein the composition is administered in the form of whitening lotion, lotion, serum, sunscreen lotion, barrier cream, mask.