Uses of quercetin-3-O-(6''-caffeoyl)-β-D-glucoside, a flavonoid compound

Abstract

The present invention relates to the use of quercetin-3-O-(6"-caffeoyl)-β-D-glucoside, a flavonoid compound. Preliminary experiments have demonstrated the use of quercetin-3-O-(6"-caffeoyl)-β-D-glucoside, a flavonoid compound described in the present invention, in the manufacture of a drug for treating vitiligo. The compound can significantly promote the effect of melanin synthesis, and is superior to the positive control drug 8-methoxypsoralen at the same dose, providing a new medicinal active compound for the treatment of vitiligo, with good application prospects.

Description

[Technical Field]

[0001] The present invention relates to the field of medicinal chemistry, in particular to the use of quercetin-3-O-(6"-caffeoyl)-β-D-glucoside, a flavonoid compound, in particular in the manufacture of drugs for the treatment of vitiligo, and belongs to the field of ethnomedicine technology. [Background technology]

[0002] Vitiligo: Vitiligo is a chronic depigmentation disease in which the epidermis, mucous membranes, and other areas lose pigment, forming white patches. The pathogenic mechanism is still unknown, but the combined effects of various factors may lead to increased destruction of skin melanocytes and reduced melanin production, thereby inducing vitiligo. Pathological characteristics of vitiligo tissue include a reduction or disappearance of melanocytes in damaged skin areas and a lack of melanocytes within cells.

[0003] The incidence of vitiligo is approximately 1%, and the prevalence does not differ by gender, race, or geographic region. Vitiligo is not contagious, but it can easily cause psychological disorders such as anxiety and depression in patients, as it affects their appearance. Currently, vitiligo treatments mainly involve drugs and ultraviolet light to stimulate localized abnormal melanocytes, promoting melanocyte regeneration and melanin production, thereby alleviating the worsening of the condition. However, the treatment effect is not very good, and the recurrence rate after stopping the medication is relatively high. Therefore, the development of a new anti-vitiligo drug with significant activity and better therapeutic effect is of great significance.

[0004] Flavonoid compounds possess a wide range of biochemical and pharmacological properties, including antioxidant, neuroprotective, anticancer, and antiallergic properties, making them attractive natural compounds for medical healthcare applications. Due to their pharmacological activity, some flavonoid compounds have been developed as health and therapeutic agents. Many flavonoids have been reported to regulate melanin production. However, although the structures of these different flavonoid compounds are similar, they exhibit both positive and negative regulatory effects on melanin production.

[0005] Quercetin-3-O-(6''-caffeoyl)-β-D-glucoside is a flavonoid compound (abbreviated as CC7) that was first isolated from Conyza filaginoides (DC) Hieron (Asteraceae), a plant of the genus Artemisia. Its antiamoebic activity has been reported. However, there have been no reports of the activity of quercetin-3-O-(6''-caffeoyl)-β-D-glucoside against vitiligo. Summary of the Invention [Means for solving the problem]

[0006] The present invention provides a use of quercetin-3-O-(6"-caffeoyl)-β-D-glucoside, a flavonoid compound. The compound promotes melanin production in skin melanocytes, and is used in the manufacture of a drug for treating vitiligo. In vitro activity tests have shown that the compound has a significant therapeutic effect on vitiligo, superior to the positive control drug 8-methoxypsoralen at the same dose, providing a new medicinally active compound for the treatment of vitiligo.

[0007] The present invention relates to a use of quercetin-3-O-(6"-caffeoyl)-β-D-glucoside, a flavonoid compound, in the manufacture of a medicament for treating vitiligo.

[0008] The compounds are used to stimulate the production of melanin in skin melanocytes.

[0009] The compounds are used to enhance tyrosinase activity.

[0010] The present invention further provides a pharmaceutical composition comprising the flavonoid compound quercetin-3-O-(6"-caffeoyl)-β-D-glucoside and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition further comprises another drug for treating vitiligo.

[0011] The present invention relates to the use of quercetin-3-O-(6″-caffeoyl)-β-D-glucoside, a flavonoid compound, the structural formula of which is as follows: [ka]

[0012] The molecular weight of quercetin-3-O-(6''-caffeoyl)-β-D-glucoside, a flavonoid compound according to the present invention, is determined by high-resolution mass spectrometry (HR-ESI-MS). The molecular ion peak is m / z [M+H] + 627.1344 (theoretical value is 627.1351), as originally reported 1 H and 13 The molecular formula of the compound is C, which is consistent with the results of the nuclear magnetic data of C. 30 H 26 O 15 It was.

[0013] The flavonoid compound quercetin-3-O-(6''-caffeoyl)-β-D-glucoside (CC7) described in this invention has excellent melanin production promoting activity and, in in vitro activity tests, has a significant vitiligo treatment effect, which is superior to the positive control drug 8-methoxypsoralen at the same dose, providing a new medicinal active compound for vitiligo treatment. [Brief explanation of the drawings]

[0014] [Figure 1] FIG. 1 is a diagram showing the effect of compounds according to the present invention on the viability of B16 mouse melanoma cells. [Figure 2] 1 is a graph showing the effect of the compounds described in the present invention on the melanin content of B16 mouse melanoma cells, where the ratios with the blank control group are **P<0.01, ****P<0.0001, and ****<0.0001. [Figure 3] 1 is a graph showing the effect of the compounds according to the present invention on the tyrosinase activity of B16 mouse melanoma cells, where **P<0.01, ***P<0.001, ***<0.001 compared to the blank control group. DETAILED DESCRIPTION OF THE INVENTION

[0015] The following examples illustrate some methods for preparing the compounds. It should be understood that the following methods, as well as other methods known to those skilled in the art, are applicable to the preparation of all compounds described in this invention. The examples are intended to illustrate, but not limit, the scope of the invention. Nuclear magnetic resonance hydrogen spectra of the compounds were measured using a Bruker ARX-400 or Bruker ARX-600, mass spectra were measured using an Agilent 1100 LC / MSD, and all reagents used were analytically pure or chemically pure. Example 1

[0016] Use of the flavonoid compound quercetin-3-O-(6''-caffeoyl)-β-D-glucoside, and measurement and evaluation of its B16 cytotoxicity compound using the CCK-8 method:

[0017] Quercetin-3-O-(6''-caffeoyl)-β-D-glucoside is obtained by the method reported by Calzada, F. et al. (Calzada, F.; Cedillo-Rivera, R.; Mata, R. Antiprotozoal activity of the constituents of Conyza filaginoides. J. Nat. Prod. 2001, 64, 671-673).

[0018] Mouse melanoma B16 cells were purchased from the Cell Bank of the Committee for the Trusteeship of Typical Cultures, Chinese Academy of Sciences (Cat# TCM2). After passage, B16 cells were cultured in a high-sugar cell medium containing 10% fetal bovine serum (FBS). 2 × 10 5Cells were seeded at 1 cell / well in a 96-well plate and cultured for 24 h. The flavonoid compound quercetin-3-O-(6'-caffeoyl)-β-D-glucoside was added at 1-50 μM to the 96-well plate and treated for 48 h. Cell viability was measured using the CCK-8 method, following the published experimental method by Win, NN, et al. (Win, NN; Woo, S.-Y.; Ngwe, H.; Wong, C.P.; Ito, T.; Okamoto, Y.; Tanaka, M.; Imagawa, H.; Asakawa, Y.; Abe, I.; et al. Tetrahydrofuran lignans: Melanogenesis inhibitors from Premna integrifolia wood collected in Myanmar. Fitoterapia 2018, 127, 308-313). The results are shown in Table 1 and Figure 1. [Table 1]

[0019] Table 1 reveals the following: Regarding the effect of 1 to 50 μM of the flavonoid compound quercetin-3-O-(6''-caffeoyl)-β-D-glucoside (CC7) on the viability of B16 cells, CC7 at concentrations of 1 to 50 μM was not toxic to the cells, indicating that the effect of CC7 at concentrations of 1 to 50 μM on B16 cells is within a safe concentration range. Example 2

[0020] Use of the flavonoid compound quercetin-3-O-(6''-caffeoyl)-β-D-glucoside to measure intracellular melanin content in B16 mouse melanoma cells:

[0021] Mouse melanoma B16 cells were subcultured and then cultured in a high-sugar cell culture medium containing 10% fetal bovine serum (FBS). 5B16 cells were seeded at 1 cell / well in 6-well plates and cultured for 24 hours. After treatment with 1-50 μM CC7 or 8-methoxypsoralen (8-MOP), the cells were cultured for 48 hours and then collected. The cells were lysed in 100 μL of RIPA cell lysis solution. After centrifugation, the supernatant was removed and 200 μL of 1 M sodium hydroxide (NaOH) lysis solution containing 10% dimethyl sulfoxide (DMSO) was added. The cells were incubated at 80°C for 1 hour. The absorbance was measured at 405 nm. Each set was repeated three times. The relative melanin content and melanogenesis activation rate were calculated. 8-Methoxypsoralen was purchased from Sigma, USA (CAS: 298-81-7).

[0022] Results: Compared with the blank control group, the melanin content of the cells in the group treated with the compounds of the present invention was significantly increased, and the results are shown in Table 2 and FIG. [Table 2]

[0023] Table 2 reveals that the melanin content of the flavonoid compound quercetin-3-O-(6"-caffeoyl)-β-D-glucoside group significantly increased at concentrations between 1 and 50 μM, demonstrating that the flavonoid compound quercetin-3-O-(6"-caffeoyl)-β-D-glucoside significantly promoted melanin production. At a concentration of 10 μM, the melanin content reached 254.9%, which was higher than the activation rate of the positive control drug 8-methoxypsoralen (8-MOP) at 50 μM. The activation rate also increased with increasing concentration. At a concentration of 50 μM, the activation rate relative to the melanin content reached 436.3%, demonstrating significant melanin production promotion activity. Example 3

[0024] Measurement of intracellular tyrosinase activity in B16 mouse melanoma cells by the flavonoid compound quercetin-3-O-(6''-caffeoyl)-β-D-glucoside:

[0025] Mouse skin melanoma B16F10 cells were subcultured and then cultured in a high-sugar cell culture medium containing 10% fetal bovine serum (FBS). 5 B16 cells were seeded at 1 cell / well in a 6-well plate and cultured for 24 h. After treatment with CC7 or 8-methoxypsoralen, samples were collected after 24 h of culture and lysed in PBS buffer (pH 6.8) containing 1% Triton X-100 and 1% sodium deoxycholate for 30 min. 10 μL of the lysate was added to 10 μL of 10 mM levodopa (L-DOPA) (CAS: 59-92-7) and incubated at 37°C for 30 min in the dark. Measurements were taken at 475 nm to calculate the relative activity and activation rate of tyrosinase. The results are shown in Table 3 and Figure 3. [Table 3]

[0026] Table 3 shows that the tyrosinase activity of the flavonoid compound quercetin-3-O-(6"-caffeoyl)-β-D-glucoside group at concentrations of 1 to 50 μM was significantly increased. The flavonoid compound quercetin-3-O-(6"-caffeoyl)-β-D-glucoside significantly regulated tyrosinase activity. At a concentration of 1 μM, the tyrosinase activity was 125.1%, which was equivalent to the activation rate of the positive control drug 8-methoxypsoralen at 50 μM. Moreover, the activation rate increased with increasing concentration. At a concentration of 50 μM, the tyrosinase activation rate reached 135.9%, significantly regulating tyrosinase activity.

[0027] Those skilled in the art will understand that although the present invention has been specifically described with reference to the above embodiments, the present invention is not limited to these specific embodiments. Based on the methods and technical solutions taught by the present invention, those skilled in the art can make appropriate modifications or improvements without departing from the spirit of the present invention, and all equivalent implementations obtained thereby fall within the scope of the present invention.

Claims

1. Use of quercetin-3-O-(6''-caffeoyl)-β-D-glucoside, a flavonoid compound, in the manufacture of a medicament for treating vitiligo.

2. 2. The use according to claim 1, wherein the compound is used to stimulate the production of melanin in skin melanocytes.

3. The use according to claim 1, characterized in that the compound is used to enhance tyrosinase activity.

4. A pharmaceutical composition for treating vitiligo, comprising quercetin-3-O-(6'-caffeoyl)-β-D-glucoside, a flavonoid compound, and a pharmaceutically acceptable carrier.

5. The pharmaceutical composition for treating vitiligo according to claim 4, further comprising another drug for treating vitiligo.

6. Use of quercetin-3-O-(6'-caffeoyl)-β-D-glucoside, a flavonoid compound, for promoting melanin production in skin melanocytes.

7. Use of quercetin-3-O-(6'-caffeoyl)-β-D-glucoside, a flavonoid compound, for improving tyrosinase activity.

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