Synergistic chloasma-removing pharmaceutical composition

Abstract

The invention discloses a pharmaceutical composition for removing chloasma with a synergistic effect. The pharmaceutical composition comprises ginsenoside Rk3 and hydroxysafflor yellow A which are used as active ingredients, and a pharmaceutically acceptable carrier, and the weight ratio of the ginsenoside Rk3 to the hydroxysafflor yellow A is 1: (0.1-0.5). Experimental results show that the improvement effect of the pharmaceutical composition on the level of tyrosinase in mouse skin is obviously improved compared with that of a single ginsenoside group Rk3 and a single hydroxysafflor yellow A group, the combination index (CI) of the ginsenoside Rk3 and the hydroxysafflor yellow A is smaller than 1, and it is shown that the combined use of the two drugs has a synergistic effect.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a pharmaceutical composition comprising ginsenoside Rk3 and hydroxysafflor yellow A and having a synergistic effect of removing chloasma. Background Art

[0002] Chloasma is a kind of light brown to brown-black flaky pigmentation spots, usually symmetrically distributed on the cheeks, nose, zygomatic region, etc., with no scales on the surface, and most patients do not feel discomfort, but the disease has a long course and is stubborn, which has a great impact on the patient's physiology and psychology. Chloasma is common in young and middle-aged women. With the pursuit of beauty by modern women, it has become particularly urgent to remove and prevent the formation of chloasma. Modern medical research believes that chloasma is mainly caused by the increase of melanocyte activity and melanin production in the skin. It is believed that the treatment of chloasma must remove the pigment in the skin and inhibit the production of melanin by reducing the activity of tyrosinase. Tyrosinase is synthesized by melanocytes. It is the key and only enzyme in the melanin synthesis process. It is mostly present in melanocytes, catalyzing the production of melanosomes, and excreting them into epidermal cells, causing the skin to produce dark brown pigmentation. Therefore, controlling the activity of tyrosinase can control the production of melanin. The activity level of tyrosinase in the body can be used as a key indicator for evaluating the condition of chloasma.

[0003] Ginsenoside Rk3 can be obtained by transformation and extraction of the dried roots and rhizomes of Panax notoginseng (Burk.) FHChen and Panax ginseng CAMey., and hydroxysafflor yellow A can be obtained by extraction of the dried flowers of Carthamus tinctorius L., a plant of the Asteraceae family.

[0004] Currently, the treatment methods for removing chloasma are divided into traditional Chinese medicine, Western medicine and integrated Chinese and Western medicine. Most of them are Western medicine compounds or Chinese herbal medicines. There are no reports on the combination of monomeric substances from different types of plants for removing chloasma and their synergistic effects. Summary of the invention

[0005] In view of the technical problems existing in the above prior art, the object of the present invention is to provide a pharmaceutical composition having a synergistic effect in removing chloasma. In addition, another object of the present invention is to provide the use of the above pharmaceutical composition in the preparation of a drug for removing chloasma.

[0006] The inventors have found through in-depth experimental studies that ginsenoside Rk3 and hydroxysafflor yellow A in a certain proportion range in the composition have a synergistic effect of removing chloasma, thereby completing the present invention. Specifically, the inventors used ginsenoside Rk3 and hydroxysafflor yellow A in different proportions to verify the chloasma removal function of chloasma model mice, and evaluated the effect of the composition on removing chloasma by detecting the activity of tyrosinase in the skin tissue of chloasma model mice at the end of administration. The experimental results showed that the regulatory effect of ginsenoside Rk3 and hydroxysafflor yellow A combined composition on the tyrosinase activity level of chloasma model mice was significantly better than that of each separate treatment group, showing that the combined use of the two has a strong synergistic effect. Therefore, the combination of ginsenoside Rk3 and hydroxysafflor yellow A has a superior effect of removing chloasma than other single-administration groups. Then, the inventors further studied the chloasma removal effect of a certain proportion of ginsenoside Rk3 and hydroxysafflor yellow A composition, and finally confirmed that it has a good chloasma removal effect.

[0007] That is, the present invention includes:

[0008] 1. A pharmaceutical composition having the effect of removing chloasma (the pharmaceutical composition of the present invention), comprising as active ingredients a therapeutically effective amount of ginsenoside Rk3 and hydroxysafflor yellow A, and a pharmaceutically acceptable carrier,

[0009] Among them, in the pharmaceutical composition, the weight ratio of ginsenoside Rk3 and hydroxysafflor yellow A is: 1:(0.1~0.5).

[0010] 2. The aforementioned pharmaceutical composition, wherein the weight ratio of ginsenoside Rk3 to hydroxysafflor yellow A is: 1:(0.2-0.4).

[0011] 3. The aforementioned pharmaceutical composition, wherein the pharmaceutical composition does not contain other saponins and / or safflower yellow.

[0012] 4. The aforementioned pharmaceutical composition, wherein the pharmaceutical composition contains ginsenoside Rk3 and hydroxysafflor yellow A as the only active ingredients for removing chloasma.

[0013] 5. Use of a composition comprising ginsenoside Rk3 and hydroxysafflor yellow A in the preparation of a chloasma-removing drug, wherein:

[0014] The weight ratio of ginsenoside Rk3 and hydroxysafflor yellow A in the composition is: 1:(0.1-0.5);

[0015] The chloasma-removing drug contains ginsenoside Rk3 and hydroxysafflor yellow A as active ingredients.

[0016] 6. The aforementioned use, wherein in the composition, the weight ratio of ginsenoside Rk3 to hydroxysafflor yellow A is: 1:(0.2-0.4).

[0017] 7. The aforementioned use, wherein the chloasma removing drug does not contain other saponins and / or safflower yellow pigment.

[0018] 8. The aforementioned use, wherein the chloasma-removing drug contains ginsenoside Rk3 and hydroxysafflor yellow A as the only active ingredients for removing chloasma.

[0019] The pharmaceutical composition of the present invention may be, for example, an oral preparation or an injection. The oral preparation may be, for example, a hard capsule, a soft capsule, a sustained-release capsule, a sugar-coated tablet, a powder, a granule, a dripping pill, a honeyed pill, a syrup or an oral liquid; the injection may be a solution, a suspension, an emulsion or a lyophilized powder. The pharmaceutical composition of the present invention may contain an excipient or other pharmaceutically acceptable carrier. The excipient may be, for example, one or more of sodium hyaluronate, sodium alginate, chitosan or collagen. DETAILED DESCRIPTION

[0020] The present invention is described in detail below in conjunction with specific embodiments. In the absence of conflict, the scientific terms in this specification have the meanings commonly understood by those skilled in the art. If there is a conflict, the definitions in this specification shall prevail.

[0021] In the present specification, ginsenoside Rk3 refers to the compound described in the following Chemical Formula 1.

[0022]

Chemical formula 1

[0023] The above-mentioned ginsenoside Rk3 is a known compound and can be prepared by methods known in the technical field. For example, ginsenoside Rk3 can be prepared by transforming, separating and purifying the total saponins in Panax notoginseng or ginseng extract.

[0024] In the present specification, hydroxysafflor yellow A refers to the compound described in the following Chemical Formula 2.

[0025]

Chemical formula 2

[0026]

[0027] The above-mentioned hydroxysafflor yellow A is a known compound and can be prepared by methods known in the art. For example, hydroxysafflor yellow A can be prepared by isolating and purifying safflor yellow in safflower extract.

[0028] The inventors have found that the combination of ginsenoside Rk3 and hydroxysafflor yellow A in the ratio range defined by the present invention not only shows synergistic effect in removing chloasma (CI value is less than 1, preferably CI value is less than 0.9), but also shows low toxicity. The inventors have also found that if the ginsenoside Rk3 and hydroxysafflor yellow A in the pharmaceutical composition are not within the ratio range defined by the present invention, there is no synergistic effect. From the perspective of significant synergistic effect (CI value is less than 0.9), in the composition, the weight ratio of ginsenoside Rk3 and hydroxysafflor yellow A is preferably 1: (0.2-0.4), for example, it can be 1: 0.3.

[0029] The pharmaceutical composition of the present invention may contain other ginsenosides and / or safflower yellow pigment, or may not contain other ginsenosides and / or safflower yellow pigment. In the case where other active pharmaceutical ingredients are contained in the pharmaceutical composition of the present invention, from the perspective of better exerting a synergistic effect, the content of the ginsenoside Rk3 and hydroxysafflor yellow pigment A is preferably 50 parts by weight or more, more preferably 60 parts by weight or more, more preferably 70 parts by weight or more, more preferably 80 parts by weight or more, more preferably 90 parts by weight or more, more preferably 95 parts by weight or more, more preferably 99 parts by weight or more, more preferably 100 parts by weight (i.e., the pharmaceutical composition contains only these two active pharmaceutical ingredients), based on the total active pharmaceutical ingredients contained in the pharmaceutical composition as 100 parts by weight. The purity of the ginsenoside Rk3 used in the pharmaceutical composition of the present invention can be 98% or more, and the purity of the hydroxysafflor yellow pigment A can be 98% or more.

[0030] The pharmaceutical composition of the present invention may contain other active ingredients for removing chloasma, or may not contain active ingredients for removing chloasma (ie, using ginsenoside Rk3 and hydroxysafflor yellow A as the only active ingredients for removing chloasma).

[0031] On the other hand, the inventors found that the pharmaceutical composition of the present invention has a significant effect in removing chloasma. Therefore, the present invention also provides the use of the pharmaceutical composition of the present invention in preparing a drug for removing chloasma.

[0032] The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier, such as an adjuvant. There are no particular restrictions on the adjuvant in the pharmaceutical composition of the present invention, and for example, adjuvants commonly used in medicines or health products in the art can be used. Specifically, the adjuvant is starch, dextrin, lactose, mannitol, sodium hydroxypropyl methylcellulose, xanthan gum, protein sugar, etc.

[0033] There is no particular limitation on the dosage form of the pharmaceutical composition of the present invention, for example, it may be an oral dosage form or an injection dosage form. The oral dosage form may be a liquid dosage form or a solid dosage form. The oral dosage form may be, for example, a hard capsule, a soft capsule, a sustained-release capsule, a compressed tablet, a sugar-coated tablet, a powder, a granule, a dripping pill, a honeyed pill, a syrup or an oral liquid; the injection dosage form may be, for example, a solution, a suspension, an emulsion or a lyophilized powder. The administration method of the pharmaceutical composition for treating insomnia may be, for example, oral administration, dripping or injection.

[0034] When preparing solid preparations for oral administration, excipients and, if necessary, binders, disintegrants, lubricants, colorants, flavoring agents, etc. can be added to the main drug, and then the mixture can be made into tablets, coated tablets, granules, fine granules, powders, capsules, etc. according to conventional methods.

[0035] As excipients, lactose, corn starch, white sugar, glucose, sorbitol, crystalline cellulose, silicon dioxide, etc. can be used; as binders, polyvinyl alcohol, ethyl cellulose, methyl cellulose, gum arabic, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, etc. can be used; as lubricants, magnesium stearate, talc, silicon dioxide, etc. can be used; as colorants, colorants allowed to be added to medicines can be used; as flavoring agents, cocoa powder, menthol, aromatic acid, peppermint oil, borneol, cinnamon powder can be used. Of course, the above-mentioned tablets and granules can also be coated with sugar coating, gelatin coating, and other necessary coatings.

[0036] When preparing injections, pH regulators, buffers, suspending aids, solubilizers, stabilizers, isotonic agents, preservatives, etc. can be added to the main drug as needed, and then prepared into intravenous, subcutaneous, or intramuscular injections according to conventional methods. At this time, a lyophilized product can also be prepared according to conventional methods as needed.

[0037] Examples of the suspending agent include methylcellulose, Tween 80, hydroxyethylcellulose, gum arabic, tragacanth gum powder, sodium carboxymethylcellulose, and polyoxyethylene sorbitan monolaurate.

[0038] Examples of the solubilizing agent include polyoxyethylene hydrogenated castor oil, Tween 80, niacinamide, polyoxyethylene sorbitan monolaurate, polyethylene glycol, and castor oil fatty acid ethyl ester.

[0039] In addition, examples of stabilizers include sodium sulfite and sodium metasulfite, and examples of preservatives include methyl parahydroxybenzoate, ethyl parahydroxybenzoate, sorbic acid, phenol, cresol, and chlorocresol.

[0040] The pharmaceutical composition of the present invention can be applied to a subject to remove chloasma. The subject can be a mammal, such as a human, rat, rabbit, sheep, pig, cow, cat, dog, monkey, etc., preferably a human.

[0041] The pharmaceutical composition of the present invention can be administered orally or parenterally. The dosage varies depending on the degree of symptoms, patient age, sex, weight, sensitivity, method of administration, period of administration, interval of administration, nature of the pharmaceutical preparation, type of active ingredient, etc., and is not particularly limited, but generally, an adult (weight 60 kg) is 1 μg to 30,000 mg per day, preferably 10 μg to 3,000 mg, more preferably 100 μg to 2,000 mg, more preferably 1 mg to 1,000 mg, more preferably 10 mg to 500 mg, more preferably 100 mg to 300 mg (based on the total amount of ginsenoside Rk3 and hydroxysafflor yellow A), for example, 15,000 mg to 50,000 mg, and the above dosage can usually be administered 1 to 3 times per day.

[0042] Example

[0043] Example 1 Effects of ginsenoside Rk3 and hydroxysafflor yellow A on tyrosinase activity in liver tissue of chloasma model mice

[0044] The compositions were prepared by mixing ginsenoside Rk3 and hydroxysafflor yellow A in weight ratios of 1:0.05, 1:0.2, 1:0.4, and 1:0.7, respectively, and were called Composition 1, Composition 2, Composition 3, and Composition 4. A group receiving separate administration of ginsenoside Rk3 and hydroxysafflor yellow A was set up as a control.

[0045] Using the above-mentioned ginsenoside Rk3, hydroxysafflor yellow A and compositions 1-4, an animal experiment on removing chloasma in chloasma model mice was conducted.

[0046] Animal source: 110 female Kunming mice, weighing 18-22 g, provided by the Animal Center of Xi'an Jiaotong University, and observed for 3 days before the experiment.

[0047] Dose selection: There were 11 groups in the experiment, namely ginsenoside Rk3 group, hydroxysafflor yellow A group 1, hydroxysafflor yellow A group 2, hydroxysafflor yellow A group 3, hydroxysafflor yellow A group 4, combination groups 1-4, control group, and model group, with 10 mice in each group. The dosage of each medication group is shown in Table 1. The control group and the model group were given an equal volume of blank solution, and the gavage volume of each mouse was 0.2 ml / 10 g, once a day, for 30 consecutive days.

[0048] Modeling method: Referring to the method reported in the literature (Miao Mingsan, Fu Zhenna, Cao Lihua. Chloasma animal model preparation specification (draft) [J]. Chinese Medicine New Drugs and Clinical Pharmacology, 2017, 28 (5): 649-653.), the hair on the back of the mouse was shaved cleanly with an electric shaver and 8% Na2S aqueous solution, so that the back skin of about 3cm×3cm was fully exposed, and the treatment was performed once every 3 days. The blank control group was injected with normal saline, and the other groups of mice were injected with progesterone at a dose of 20mg / kg per day (the injection site was the muscle at the root of the mouse hind limb, and the injection was alternated on both sides). At the same time, the skin of the back of the other groups of mice was irradiated with medium-wave ultraviolet light (320nm), the distance of the light source was 20cm, and each irradiation was 30min, once a day, for 30 consecutive days.

[0049] Index detection: 1 hour after the last administration, take an appropriate amount of skin from the ultraviolet irradiated area on the back of the mouse, make a 10% homogenate in an ice bath, and use the tyrosinase catalytic oxidation of levodopa (L-DOPA) rate method to detect the activity of tyrosinase in the skin tissue (Lou Caixia, Tian Yanze, Pu Xianglan. Study on the inhibitory effect of different polar parts of Forsythia suspensa on tyrosinase activity [J]. Lishizhen Traditional Chinese Medicine, 2011, 22(10): 2415-2416.).

[0050] The drug combination index of ginsenoside Rk3 / hydroxysafflor yellow A was calculated as CI = (A × B) / AB (Romanelli S, Perego P, Pratesi G, Carenini N, Tortoreto M, Zunino F. In vitro and in vivo interaction between cisplatin and topotecan in ovarian carcinoma systems. [J]. Cancer chemotherapy and pharmacology, 1998, 41 (5): 385-390). When CI < 1, it indicates that the combination has a synergistic effect; when CI ≤ 0.9, it indicates that the synergistic effect is significant. Among them, C = tyrosinase activity of the model group - tyrosinase activity of the control group, representing the lower limit of the reduction of tyrosinase activity after administration;

[0051] T = tyrosinase activity in each drug-administered group minus tyrosinase activity in the control group, representing the actual level of tyrosinase activity reduction after drug administration;

[0052] A and B represent the (1-T / C) values ​​when ginsenoside Rk3 or hydroxysafflor yellow A is administered alone, respectively; AB represents the (1-T / C) values ​​when combination 1-4 is administered.

[0053] The results are shown in Table 1 below:

[0054] Table 1 Effects of drug administration on tyrosinase activity in skin tissue of chloasma model mice

[0055]

[0056] Note: The dosages of compositions 1-4 are the dosages of ginsenoside Rk3 group combined with hydroxysafflor yellow A groups 1-4, respectively.

[0057] As can be seen from Table 1, after oral administration of different groups of drugs to chloasma model mice for 30 days, the drug combination index CI values ​​of composition 2, composition 3 and ginsenoside Rk3 group, as well as the corresponding dose of hydroxysafflor yellow A group, were all less than 0.9, but the medicinal combination index CI values ​​of composition 1 and composition 4 were greater than 1, indicating that ginsenoside Rk3: hydroxysafflor yellow A can synergistically reduce the tyrosinase activity of skin tissue in chloasma model mice in the range of 1:0.2-0.4, thereby exerting the effect of removing chloasma, but there is no synergistic effect outside the above range.

[0058] Therefore, it can be considered that the combination of ginsenoside Rk3 and hydroxysafflor yellow A in a certain ratio can achieve the purpose of removing or improving chloasma by downregulating the tyrosinase level of skin tissue. At the same time, in all animal experiments involved in this embodiment, the animals in each group moved freely, without abnormal body weight, food intake and diet compared with the control group. There was no statistical difference, which shows that the combination of ginsenoside Rk3 and hydroxysafflor yellow A is safe.

[0059] Example 2 Trial test of the efficacy of a chloasma removal composition

[0060] Composition 3 in Example 1 was selected to test its efficacy in removing chloasma on human body.

[0061] 1. Materials and Methods

[0062] 1.1 Subjects

[0063] Age 18-65, qualified in physical examination, willing to participate in the test, with facial melasma ranging from light brown to dark brown, with clear boundaries, symmetrical distribution, and no inflammatory manifestations and scaling. The following conditions are excluded: pregnant or lactating women, allergic constitution; those with serious diseases such as cardiovascular, cerebrovascular, liver, kidney and hematopoietic system; those who did not take the test sample according to the regulations, and the efficacy cannot be determined or the incomplete data affects the judgment of efficacy or safety.

[0064] 1.2 Experimental design and grouping

[0065] 100 voluntary subjects were randomly divided into a trial group and a control group according to the color and area of ​​chloasma, with 50 subjects in each group. The trial group took Composition 3 at 200 mg / day, and the control group was administered with vitamin C and vitamin E, vitamin C 0.2g / time, 1 time / d, vitamin E 0.1g / time, 1 time / d. The test period was 60 days. During the test, the original eating habits were not changed, and the subjects had a normal diet. The subjects should pay attention to sun protection and take physical sun protection measures.

[0066] 2. Observation indicators

[0067] 2.1 Efficacy indicators and efficacy evaluation criteria

[0068] The efficacy indicators are as follows: The Melasma Area and Severity (MASI) score is the main tool for evaluating the treatment effect of melasma, including the evaluation of the area, color depth, and color uniformity of the forehead, right zygomatic region, left zygomatic region, and chin of the subjects. The corresponding scores are 0 to 4 points, the total score is 48 points, and the lowest score is 0. [Pigmentary Disease Working Group of the Dermatology Branch of the Chinese Medical Doctor Association. Chinese Expert Consensus on the Diagnosis and Treatment of Melasma (2021 Edition) [J]. Chinese Journal of Dermatology, 2021, 54(2): 110-115.].

[0069] The efficacy evaluation criteria are as follows: basically cured: the percentage of improvement of the efficacy index is ≥80%; markedly effective: the percentage of improvement of the efficacy index is 50% to 79%; improved: the percentage of improvement of the efficacy index is 10% to 49%; ineffective: the percentage of improvement of the efficacy index is <10%. Effective rate = (the number of basically cured cases + the number of markedly effective cases) / the total number of cases × 100% [Amit GP, Linda SH, Rafia B, et al. Reliability assessment and validation of the Melasma Area and Severity Index (MASI) and a new modified MASI scoring method [J]. Journal of the American Academy of Dermatology, 2009, 64 (1): 78-83.].

[0070] 3. Treatment results before and after the trial

[0071] After 60 days of trial eating, the treatment effect of melasma in each group of patients is shown in Table 2 below:

[0072] Table 2 Comparison of therapeutic effects between the trial group and the control group

[0073]

[0074] As shown in Table 2, after 60 days of treatment, the effective rate of the control group was 42.00%, and the effective rate of the trial group was 72.00%, which further illustrates the synergistic effect of ginsenoside Rk3 group and hydroxysafflor yellow A in removing chloasma.

[0075] It should be noted that, under the premise that it is feasible and does not obviously violate the main purpose of the present invention, any technical feature or combination of technical features described as a component of a technical solution in this specification can also be applied to other technical solutions; and, under the premise that it is feasible and does not obviously violate the main purpose of the present invention, the technical features described as components of different technical solutions can also be combined in any way to form other technical solutions. The present invention also includes technical solutions obtained by combination under the above circumstances, and these technical solutions are equivalent to those recorded in this specification.

[0076] The present invention is described above through specific implementation modes and examples, but those skilled in the art should understand that these are not intended to limit the scope of the present invention, and the scope of the present invention should be determined by the claims.

[0077] Industrial Applicability

[0078] The present invention provides a pharmaceutical composition comprising ginsenoside Rk3 and hydroxysafflor yellow A and having a synergistic effect of removing chloasma.

Claims

1. A pharmaceutical composition having the effect of removing chloasma, comprising ginsenoside Rk3 and hydroxysafflor yellow A as active ingredients, and a pharmaceutically acceptable carrier, in, In the pharmaceutical composition, the weight ratio of ginsenoside Rk3 to hydroxysafflor yellow A is 1:(0.1-0.5).

2. The pharmaceutical composition according to claim 1, wherein In the pharmaceutical composition, the weight ratio of ginsenoside Rk3 to hydroxysafflor yellow A is 1:(0.2-0.4).

3. The pharmaceutical composition according to claim 1, wherein The pharmaceutical composition does not contain other saponins and / or safflower yellow pigment.

4. The pharmaceutical composition according to claim 1, wherein The pharmaceutical composition contains ginsenoside Rk3 and hydroxysafflor yellow A as the only active ingredients for removing chloasma.

5. Use of a composition comprising ginsenoside Rk3 and hydroxysafflor yellow A in the preparation of a chloasma-removing drug, wherein: The weight ratio of ginsenoside Rk3 and hydroxysafflor yellow A in the composition is: 1:(0.1-0.5); The chloasma-removing drug contains ginsenoside Rk3 and hydroxysafflor yellow A as active ingredients.

6. The use according to claim 5, wherein In the composition, the weight ratio of ginsenoside Rk3 to hydroxysafflor yellow A is 1:(0.2-0.4).

7. The use according to claim 5, wherein The chloasma removing drug does not contain other saponins and / or safflower yellow pigment.

8. The use according to claim 5, wherein The chloasma-removing drug contains ginsenoside Rk3 and hydroxysafflor yellow A as the only active ingredients for removing chloasma.

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