Use of a 2-mercaptobenzothiazole compound for preparing a tyrosinase inhibitor

Abstract

The application relates to application of a 2-mercaptobenzothiazole compound in preparation of a tyrosinase inhibitor. The 2-mercaptobenzothiazole compound provided by the application has excellent human tyrosinase (hTyr) inhibiting activity and oil solubility, and can be prepared into a safe and effective whitening product.

Description

Technical Field

[0001] This invention relates to the field of tyrosinase inhibitor technology, and more particularly to the application of a 2-mercaptobenzothiazole compound in the preparation of tyrosinase inhibitors. Background Technology

[0002] The melanin produced on the human body surface is mainly divided into two types: eumelanin and pheomelanin. Eumelanin primarily affects the color of skin, eyes, and hair, while pheomelanin mainly affects the lips and nipples. The melanin produced by the skin is mainly eumelanin, which protects the skin from ultraviolet radiation and prevents DNA mutations and the development of skin cancer. On the other hand, excessive melanin production can lead to pigmentation, such as freckles, age spots, melasma, moles, and sunspots, affecting appearance. Tyrosinase, also known as polyphenol oxidase, is a copper-containing oxidoreductase widely found in microorganisms, animals, and plants. Tyrosinase is produced by melanocytes. After being produced and processed in the endoplasmic reticulum and Golgi apparatus, it is transported to melanosomes, where melanin is synthesized. Tyrosinase is a key rate-limiting enzyme in the melanin synthesis pathway, directly affecting melanin synthesis. The expression and activity of tyrosinase determine the rate and yield of melanin production; the higher the tyrosinase activity, the greater the amount of melanin formed in the skin.

[0003] Numerous inhibitors targeting tyrosinase have been developed, particularly hydroquinone and its derivatives, including hydroquinone (also known as hydroquinone), arbutin, kojic acid, azelaic acid, 4-butylresorcinol, 4-hexylresorcinol, 4-phenylethylresorcinol, and dimethoxytolylpropylresorcinol. However, all these agents lack efficacy and / or safety, so currently only a few are used in clinical dermatology. For example, hydroquinone has been banned for use in skin whitening products due to its high cytotoxicity; kojic acid is sensitizing; arbutin requires a high effective concentration; and nicotinamide and its analogues have drawbacks such as teratogenicity and intolerance in some populations.

[0004] Recently, two highly effective tyrosinase inhibitors with similar molecular formulas, Isobutylamidothiazolyl resorcinol and KT-939 (Methyloxetanecarbamido Thiazolyl Resorcinol), have been invented and used in the skincare industry. These two small molecules exhibit an IC50-950 response to the human tyrosinase hTyr. 50 The concentrations are 1.1 μM and 0.06 μM respectively, but the effective addition level in skincare products is only 0.2% (approximately 6.5 mM). This is because the molecular weight is relatively large, resulting in a very low transdermal penetration rate. Furthermore, Asians tend to have thinner epidermal layers, making products containing peptides like Ambroxol more prone to causing acne, as they are extremely difficult to dissolve in skincare bases and precipitate out onto the epidermis. Therefore, when developing effective tyrosinase inhibitors for skincare products, the IC50 concentration should not be considered alone. 50It's not just one indicator. Rather, it takes into account factors such as compatibility with subsequent skincare products, transdermal absorption rate, and molecular weight. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides the application of 2-mercaptobenzothiazole compounds in the preparation of tyrosinase inhibitors. The 2-mercaptobenzothiazole compounds provided by this invention possess excellent human tyrosinase (hTyr) inhibitory activity and oil solubility, and can be used to prepare safe and effective skin-whitening products.

[0006] To achieve this objective, the present invention adopts the following technical solution: In a first aspect, the present invention provides the use of a 2-mercaptobenzothiazole compound in the preparation of a tyrosinase inhibitor, wherein the 2-mercaptobenzothiazole compound has the following structure: Formula I; Among them, R1-R4 are each independently selected from hydrogen, deuterium, fluorine, chlorine, bromine, iodine, -CN, -N(R a1 )2、-OR a2 -R a3 -NHR b1 -OR b2 ; R a1 R a2 R a3 Whether the same or different, each is independently selected from H, C1-C6 alkyl; when R a1 R a2 R a3 When it is a C1-C6 alkyl group, R1-R4 exist independently, or two adjacent groups form a 4-8 membered ring with the carbon, nitrogen or oxygen atoms they are connected to; R b1 R b2 Whether identical or different, each is independently selected from monosaccharide residues, oligosaccharide residues, -P(=O)(OM1)(OM2), and -P(=O)R. c1 R c2 ; M1 and M2 may be the same or different, and each is independently selected from H and C1-C6 alkyl groups; R c1 R c2 They may be the same or different, and each is independently selected from C1-C6 alkyl groups.

[0007] The aforementioned C1-C6 alkyl groups can be, for example, C1, C2, C3, C4, C5, or C6.

[0008] Preferably, the monosaccharide residues are selected from glucosyl, mannose, galactosyl, xylose, lysoose, fucose, arabinose, rhamnose, fructose, sorbitol, and tagatose.

[0009] Preferably, the oligosaccharide residues are selected from lactose, maltoluose, palaginose, lactulose, amygdalinose, menobiose, cellobiose, isomaltose, rutinose, and maltose.

[0010] Preferably, R1-R4 are each independently selected from hydrogen or chlorine.

[0011] Preferably, the 2-mercaptobenzothiazole compound is selected from any one of the following structures: , .

[0012] Secondly, the present invention provides the use of a 2-mercaptobenzothiazole compound in the preparation of a medicament for treating freckles, age spots, melasma, moles, and sunspots, wherein the 2-mercaptobenzothiazole compound has the following structure: ; Among them, R1-R4 are defined as the same structure as Equation I.

[0013] Preferably, R1-R4 are each independently selected from hydrogen or chlorine.

[0014] Preferably, the 2-mercaptobenzothiazole compound is selected from any one of the following structures: , .

[0015] Preferably, the drug further includes pharmaceutically acceptable excipients.

[0016] Preferably, the pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carriers, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH adjusters, antioxidants, antibacterial agents, or buffers.

[0017] Thirdly, the present invention provides the application of a 2-mercaptobenzothiazole compound in the preparation of cosmetics with whitening effects, wherein the 2-mercaptobenzothiazole compound has the following structure: ; Among them, R1-R4 are defined as the same structure as Equation I.

[0018] Preferably, R1-R4 are each independently selected from hydrogen or chlorine.

[0019] Preferably, the 2-mercaptobenzothiazole compound is selected from any one of the following structures: , .

[0020] Preferably, the cosmetic also includes cosmetically acceptable excipients; Preferably, the cosmetic excipients include any one or a combination of at least two of the following: emulsifiers, moisturizers, thickeners, skin conditioners, pH adjusters, or chelating agents.

[0021] Preferably, the cosmetics include skin care water, lotion, essential oil, cream, ointment or mask, and are preferably lotion, essential oil, cream or ointment.

[0022] Compared with the prior art, the present invention has at least the following beneficial effects: The 2-mercaptobenzothiazole compounds provided by this invention have excellent human tyrosinase inhibitory activity and oil solubility, and can be used in drugs for treating freckles, age spots, melasma, moles, and sun-induced pigmentation or in whitening cosmetics, especially suitable for oil-soluble cosmetics such as essential oils, creams, lotions, and ointments. Attached Figure Description

[0023] Figure 1 This is the absorbance graph of melanin with added compound a.

[0024] Figure 2 This is a cell diagram with compound a added.

[0025] Figure 3 This is the absorbance graph of melanin with added compound b.

[0026] Figure 4 This is a cell diagram with compound b added.

[0027] Figure 5 This is a toxicity test diagram of compounds a, b, and peptide ambidol. Detailed Implementation

[0028] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. However, the following examples are merely simplified examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention is determined by the claims.

[0029] The compounds used in the following test examples are as follows: Compound a: 2-Mercaptobenzothiazole, CAS: 149-30-4, structural formula is ; Compound b: 2-Mercapto-5-chlorobenzothiazole, CAS: 5331-91-9, structural formula is .

[0030] Test Example 1 Test of the inhibitory activity of the compound on melanin production Experimental method: 5×10 5 293T cells were seeded into 6-well plates. After 24 hours, 4 μg of plasmid overexpressing human tyrosinase hTyr was introduced. After another 24 hours, the test compound was added at final concentrations of 0, 4, 8, 20, 40, and 80 μM. L-tyrosine disodium dihydrate (0.72 mg / mL) was added to DMEM culture medium to increase melanin production. Cells were photographed and collected after 3 days of culture. Melanin was extracted using 1 mL of ethylbenzene / ethanol (50 / 50) mixture and dried. The melanin was then dissolved in 200 μL of 1 M NaOH solution heated to 85°C. The supernatant was collected and the absorbance at 405 nm was measured using a microplate reader.

[0031] After 3 days of incubation, the absorbance of the systems with different concentrations of compound a added is as follows: Figure 1 As shown, cell photographs are as follows Figure 2 As shown, the absorbance of the systems with different concentrations of compound b added is as follows: Figure 3 As shown, cell photographs are as follows Figure 4 As shown in the figure, it can be seen that the melanin synthesis decreases with increasing compound concentration in the system with added compound a or compound b, indicating that both compounds have excellent human tyrosinase inhibitory activity.

[0032] Test Example 2 Oil solubility test of compounds Compounds a, b, and peptide amecto were dissolved in coconut oil, and their solubility at 25°C was tested. The results are shown in Table 1.

[0033] Table 1 Test results show that compounds a and b provided by this invention have extremely high oil solubility compared to other tyrosinase inhibitors, making them suitable for use in oil-soluble cosmetics such as essential oils, creams, lotions, and ointments.

[0034] Test Example 3 Toxicity testing of compounds On the first day, 1×10 4 Two 293T cells were cultured in 24-well plates. On the second day, 500 μL of culture medium containing 80 μM of drug was added (DMSO was used as a negative control). Two days later, 50 μL of 5 mg / mL MTT and 450 μL of phenol red-free DMEM were added, and the cells were incubated for 3 hours (37°C). After centrifugation, the cells were dissolved in 500 μL of DMSO, and the absorbance was read at 590 nm.

[0035] Compounds a, b, and peptide amecto were tested, and the results are as follows:Figure 5 As shown, compounds a and b have better safety profiles than peptide Amidato.

[0036] The applicant declares that the above description is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention fall within the protection and disclosure scope of the present invention.

Claims

1. The application of a 2-mercaptobenzothiazole compound in the preparation of tyrosinase inhibitors, characterized in that, The 2-mercaptobenzothiazole compounds have the following structures: Formula I; Among them, R1-R4 are each independently selected from hydrogen, deuterium, fluorine, chlorine, bromine, iodine, -CN, -N(R a1 )2、-OR a2 -R a3 -NHR b1 -OR b2 ; R a1 R a2 R a3 Whether the same or different, each is independently selected from H, C1-C6 alkyl; when R a1 R a2 R a3 When it is a C1-C6 alkyl group, R1-R4 exist independently, or two adjacent groups form a 4-8 membered ring with the carbon, nitrogen or oxygen atoms they are connected to; R b1 R b2 Whether identical or different, each is independently selected from monosaccharide residues, oligosaccharide residues, -P(=O)(OM1)(OM2), and -P(=O)R. c1 R c2 ; M1 and M2 may be the same or different, and each is independently selected from H and C1-C6 alkyl groups; R c1 R c2 They may be the same or different, and each is independently selected from C1-C6 alkyl groups.

2. The application according to claim 1, characterized in that, R1-R4 are each independently selected from hydrogen or chlorine.

3. The application according to claim 1 or 2, characterized in that, The 2-mercaptobenzothiazole compounds are selected from any of the following structures: 、 。 4. The use of a 2-mercaptobenzothiazole compound in the preparation of a medicament for treating freckles, age spots, melasma, moles, and sunspots, characterized in that, The 2-mercaptobenzothiazole compounds have the following structures: ； Among them, R1-R4 are defined as the same structure as Equation I.

5. The application according to claim 4, characterized in that, R1-R4 are each independently selected from hydrogen or chlorine; Preferably, the 2-mercaptobenzothiazole compound is selected from any one of the following structures: 、 。 6. The application according to claim 4 or 5, characterized in that, The drug also includes pharmaceutically acceptable excipients; Preferably, the pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carriers, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH adjusters, antioxidants, antibacterial agents, or buffers.

7. The application of a 2-mercaptobenzothiazole compound in the preparation of cosmetics with whitening effects, characterized in that, The 2-mercaptobenzothiazole compounds have the following structures: ； Among them, R1-R4 are defined as the same structure as Equation I.

8. The application according to claim 7, characterized in that, R1-R4 are each independently selected from hydrogen or chlorine; Preferably, the 2-mercaptobenzothiazole compound is selected from any one of the following structures: 、 。 9. The application according to claim 7 or 8, characterized in that, The cosmetics also include cosmetically acceptable excipients; Preferably, the cosmetic excipients include any one or a combination of at least two of the following: emulsifiers, moisturizers, thickeners, skin conditioners, pH adjusters, or chelating agents.

10. The application according to any one of claims 7-9, characterized in that, The types of cosmetics include skin care water, lotion, essential oil, cream, ointment or mask, preferably lotion, essential oil, cream or ointment.

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