Gentiopicroside composition as well as preparation method and application thereof

Abstract

The invention relates to a gentiopicroside composition as well as a preparation method and application thereof. The gentiopicroside composition is prepared from a collagen derivative and gentiopicroside, wherein the collagen derivative is a condensation product of a compound A and collagen, and the compound A is a functional component with carboxyl. The gentiopicroside composition provided by the invention has a good permeation promoting effect and has a wide application prospect in the field of skin care products.

Description

Technical Field

[0001] The invention relates to a gentiopicroside composition and a preparation method and application thereof. Background Art

[0002] With the development of the economy, people's yearning for a better life is becoming more and more extreme, and this is accompanied by excessive skin care. This excessive skin care often causes the stratum corneum of the skin to become thinner and thinner, which makes it easier for harmful substances from the outside to penetrate the stratum corneum and enter the skin, causing skin sensitivity. Once the skin becomes sensitive, it will cause redness, swelling, pain, itching and other phenomena, seriously affecting the patient's quality of life.

[0003] TNF-α causes the expression of genes related to sensitive skin, especially in the expression of genes encoding cytokines and vascular endothelial growth factor. IL-22, produced by skin c-Kit+FcεRI+ mast cells, Th17, Th22 and Tc22 cells (CD8+T cells), is the main cytokine associated with the proliferation of keratinocytes in sensitive skin. In addition, IL-17A, SOM and IL-1α are also the main inflammatory factors that cause skin sensitivity. Therefore, improving sensitive skin by regulating the above inflammatory factors is a new way to treat skin. Summary of the invention

[0004] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a gentiopicroside composition and a preparation method and application thereof.

[0005] In a first aspect, the present invention provides a gentiopicroside composition, comprising a collagen derivative and gentiopicroside;

[0006] The collagen derivative is a condensation product of compound A and collagen, wherein compound A is an effective ingredient having a carboxyl group.

[0007] According to an embodiment of the present invention, the weight ratio of the collagen derivative to the gentiopicroside is 1:(0.01-10), for example, 1:0.01, 1:0.1, 1:0.2, 1:0.5, 1:1, 2:1, 3:1, 4:1 or 5:1.

[0008] According to an embodiment of the present invention, the collagen derivative and the gentiopicroside form nanoparticles.

[0009] According to an embodiment of the present invention, the compound A is selected from retinoic acid or its derivatives.

[0010] According to an embodiment of the present invention, the compound A is tretinoin and / or isotretinoin.

[0011] According to an embodiment of the present invention, the average molecular weight of the collagen derivative is 200-1000 KDa.

[0012] According to an embodiment of the present invention, the average molecular weight of the collagen is 500Da-1 million Da, for example, 800Da, 1000Da, 1200Da, 1400Da, 1600Da, 1800Da, 2000Da, 2500Da, 3000Da, 3500Da, 4500Da, 5500Da, 7500Da, 100000 Da, 400000 Da, or 700000 Da.

[0013] According to an embodiment of the present invention, the average molecular weight of the collagen is 500Da-300,000Da.

[0014] According to an embodiment of the present invention, the average molecular weight of the collagen is 500Da-5000Da.

[0015] According to an embodiment of the present invention, the average molecular weight of the collagen is 500-1500 Da.

[0016] According to an embodiment of the present invention, the method for preparing the collagen derivative comprises: subjecting collagen to a condensation reaction with compound A to generate the collagen derivative.

[0017] According to an embodiment of the present invention, the molar ratio of the collagen to the compound A is 1:(0.01-1); for example, 1:0.01, 1:0.1, 1:0.2, 1:0.3, 1:0.4, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1.

[0018] According to an embodiment of the present invention, the condensation reaction uses EDC and NHS as condensation reagents.

[0019] According to an embodiment of the present invention, the molar ratio of the compound A to the EDC is 1:(0.5-1.5); for example, 1:0.5, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5.

[0020] According to an embodiment of the present invention, the molar ratio of the compound A to the NHS is 1:(0.5-1.5); for example, 1:0.5, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5.

[0021] According to an embodiment of the present invention, the condensation reaction time is 0.1 to 24 hours.

[0022] According to an embodiment of the present invention, the condensation reaction time is 0.1 to 4 hours.

[0023] According to an embodiment of the present invention, the temperature of the condensation reaction is -5 to 10°C, for example, -5°C, -2°C, 0°C, 1°C, 2°C, 3°C, 4°C, 6°C, 8°C, or 10°C.

[0024] According to an embodiment of the present invention, the temperature of the condensation reaction is 0-4°C.

[0025] According to an embodiment of the present invention, the compound A is first dissolved in a mixed solution of PBS solution and tetrahydrofuran before the condensation reaction.

[0026] According to an embodiment of the present invention, the pH value of the PBS solution is 2-6, for example, 2, 3, 4, 5, 6.

[0027] According to an embodiment of the present invention, the concentration of the PBS solution is 0.05-1M, for example, 0.05M, 0.1M, 0.2M, 0.3M, 0.4M, 0.5M, 0.6M, 0.7M, 0.8M, 0.9M, 1M.

[0028] According to an embodiment of the present invention, the volume ratio of the PBS solution to tetrahydrofuran is 1:(0.5-2), for example, 1:0.5, 1:1, 1:1.3, 1:1.5, 1:1.7, or 1:2.

[0029] According to an embodiment of the present invention, the preparation method comprises: dissolving compound A in a mixed solution of a PBS solution (0.05M-1M) with a pH of 2-6 and tetrahydrofuran (1:0.5-2), adding EDC and NHS thereto, activating at 0-4°C for 0.1-4h, then adding collagen thereto, controlling the temperature to react for 0-24h; after the reaction is completed and dialyzed (MW-3000Da), a collagen derivative is obtained after freeze-drying.

[0030] In a second aspect, the present invention provides a method for preparing a gentiopicroside composition, comprising:

[0031] dissolving the collagen derivative in a solvent to obtain a solution A;

[0032] Dissolving gentiopicroside in a solvent to obtain a solution B;

[0033] Mix solution A and solution B.

[0034] According to an embodiment of the present invention, the solvent is water.

[0035] According to an embodiment of the present invention, the mass concentration of the collagen derivative in the solution A is 0.001% to 50%, for example, 0.001%, 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 1.0%, 2%, 5%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45%.

[0036] According to an embodiment of the present invention, the mass concentration of gentiopicroside in the solution B is 0.001% to 50%, for example, 0.001%, 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 1.0%, 2%, 5%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45%.

[0037] According to an embodiment of the present invention, the mixing conditions include: a stirring speed of 10 to 2000 rpm and a stirring time of 0.1 to 48 h.

[0038] In a third aspect, the present invention provides a skin care product, comprising the gentiopicroside composition described in the first aspect of the present invention or the gentiopicroside composition obtained by the preparation method described in the second aspect of the present invention.

[0039] According to an embodiment of the present invention, the functional ingredients are functional ingredients for skin, such as whitening and acne-removing ingredients, including but not limited to: tranexamic acid (tranexamic acid), niacin, niacinamide, salicylic acid, etc.

[0040] According to an embodiment of the present invention, the skin care product may be water, lotion, cream, facial mask, etc.

[0041] According to an embodiment of the present invention, the mass ratio of the gentiopicroside composition to the effective ingredient is 1:(0.01-10), for example, 1:0.01, 1:0.1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

[0042] Gentiopicroside has an obvious therapeutic effect on the repair of sensitive skin. Gentiopicroside belongs to the iridoid glycosides and is mainly found in Gentiana macrophylla and Gentiana scabra. Gentiana macrophylla and Gentiana scabra are plants of the Gentianaceae family Gentiana, and are traditional Chinese medicines. Pharmacological studies on gentiopicroside have shown that it has biological activities of antioxidant, anti-inflammatory, antibacterial, antiproliferative and liver protection, osteoarthritis, lung injury, and anti-tumor effects. More and more studies have confirmed that gentiopicroside can play an anti-inflammatory role by downregulating the expression of inflammatory factors (such as TNF-α, IL-17A, TGF-β1, IL-6, etc.). Although its therapeutic effect is good, due to its unstable physicochemical properties, poor solubility and low bioavailability, the therapeutic effect in vivo will be significantly reduced. The development of nanotechnology provides a new method for the percutaneous absorption of the above-mentioned drugs. The present invention uses collagen derivatives as a promoter for percutaneous absorption to form a composition with gentiopicroside, thereby improving the percutaneous absorption efficiency of gentiopicroside and promoting its therapeutic effect.

[0043] The gentiopicroside composition of the present invention can effectively promote the transdermal absorption of gentiopicroside and has broad application prospects in the field of skin care products. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 This is the NMR spectrum of the collagen derivative of Example 1 of the present invention;

[0045] Figure 2 is the infrared spectrum of the collagen derivative of Example 1 of the present invention;

[0046] Figure 3 This is a fluorescent imaging diagram of the animal skin test of the sample in Application Example 1 of the present invention;

[0047] Figure 4 This is the therapeutic effect of Application Example 2 of the present invention on a psoriasis animal model. DETAILED DESCRIPTION

[0048] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with the embodiments and drawings. The specific embodiments described herein are only used to explain the present invention and are not intended to constitute any limitation to the present invention. In addition, in the following description, the description of known structures and technologies is omitted to avoid unnecessary confusion of the concepts of the present disclosure. Such structures and technologies are also described in many publications.

[0049] The collagen used in the embodiments of the present invention is a hydrolyzed collagen with a molecular weight distribution of 500-1500Da or a molecular weight distribution of 2000-3000Da. The collagen in the embodiments of the present invention can be purchased.

[0050] Example 1

[0051] Take 3g of retinoic acid and dissolve it in a mixed solution of 30mL pH=5 (0.1M) PBS and 30mL tetrahydrofuran, add 2.2g of EDC and 1.6g of NHS under ice bath (0℃), react for 2 hours, then add 5g of collagen (500-1500Da), continue to react at room temperature for 12 hours, dialyze (MW=500Da), and freeze-dry to obtain 4.5g of product.

[0052] Example 2

[0053] Take 6g of retinoic acid and dissolve it in a mixed solution of 50mL pH=4 (0.1M) PBS and 50mL tetrahydrofuran, add 6g of EDC and 4.5g of NHS under ice bath (0℃), react for 1 hour, then add 15g of collagen (2000-3000Da), continue to react at room temperature for 12 hours, dialyze (MW=500Da), and freeze-dry to obtain 6.8g of product.

[0054] Example 3

[0055] Take 3g of isotretinoin and dissolve it in a mixed solution of 50mL pH=4 (0.1M) PBS and 50mL tetrahydrofuran, add 3g of EDC and 2.3g of NHS under ice bath (0℃), react for 1 hour, then add 5g of collagen (500-1500Da), continue to react at room temperature for 12 hours, dialyze (MW=500Da), and freeze-dry to obtain 4.8g of product.

[0056] Figure 1 is the hydrogen spectrum of Example 1, Figure 2 is the infrared spectrum of Example 1, Figure 1 and Figure 2 It can be seen that the collagen derivative of the present invention contains an amide bond.

[0057] Example 4

[0058] Take 1 g of the collagen derivative prepared in Example 1 and dissolve it in 100 mL of water as solution A; take 0.1 g of gentiopicroside and dissolve it in 100 mL of water as solution B; at room temperature, under stirring at 200 rpm, add solution B to solution A to prepare a gentiopicroside composition; the obtained composition can be directly used as a functional substance in skin care products after high-temperature sterilization.

[0059] Example 5

[0060] 0.5 g of the collagen derivative prepared in Example 1 was dissolved in 100 mL of water as solution A; 0.1 g of gentiopicroside was dissolved in 100 mL of water as solution B; at room temperature, solution B was added to solution A under stirring at 200 rpm to prepare a gentiopicroside composition; the obtained composition can be directly used as a functional substance in skin care products after high-temperature sterilization.

[0061] Example 6

[0062] 0.1 g of the gentiopicroside composition prepared in Example 4 was dissolved in 5 mL of aqueous solution to obtain functionalized gentiopicroside.

[0063] 30 g of glycerin, 0.5 g of ice crystals, 0.5 g of carbomer, 4 g of jojoba wax PEG-120 ester, 4 g of PEG-20 methyl glucose sesquistearate, 2 g of p-hydroxyacetophenone, and 923.9 g of deionized water were heated to 85° C. and mixed to obtain a mixture;

[0064] After the temperature drops to 45°C, the functionalized gentiopicroside is added to the mixture and stirred to fully dissolve;

[0065] Then add 10 g of 1,2-pentanediol to the above solution, stir thoroughly to obtain essence a, cool to room temperature, discharge and let stand.

[0066] Example 7

[0067] 0.5 g of the gentiopicroside composition prepared in Example 4 was dissolved in 10 ml of aqueous solution to obtain functionalized gentiopicroside.

[0068] Heat 30 g of emulsifier Emulium Delta MB, 5 g of hydrogenated lecithin, 10 g of cetearyl alcohol, 15 g of Lipocire SG, 30 g of Dow Corning PMX200-5cst, and 20 g of caprylic / capric triglyceride to 85° C. to prepare a mixture A;

[0069] Put 30g of glycerol, 3g of ice crystal AVC, 1g of xanthan gum, 6g of hexanediol, 15g of butanediol, 5g of p-hydroxyacetophenone and 830g of deionized water into a stirring pot, stir and dissolve while heating to 85-95°C to fully dissolve, and keep warm for 15-20 minutes to prepare a mixture B;

[0070] Add mixture A to mixture B and stir for 40 min to obtain mixture C;

[0071] The mixture C was cooled to 40-45° C., the functionalized gentiopicroside was added, and the mixture was stirred to dissolve to obtain a mixture D;

[0072] While maintaining the temperature at 40-45°C, add 1 g of methylparaben to mixture D and stir thoroughly to obtain emulsion c. Cool to room temperature, discharge and allow to stand.

[0073] Application Example 1

[0074] 1. Test sample preparation

[0075] A rhodamine-labeled collagen derivative (Example 1) (RhB-VA-Col) was prepared and formulated into 5 mL of a 1 wt % aqueous solution.

[0076] The sample of Example 7 was labeled with rhodamine to prepare a rhodamine-labeled gentiopicroside composition (RhB-VA-Col@Gen), and was configured into a 5 mL aqueous solution of 1 wt %.

[0077] 2. Transdermal test

[0078] 20 C57BL / 6 mice were randomly divided into 2 groups, 10 mice in each group, that is, 2 pieces of 5*5cm hair of each mouse were removed with an electric shaver to expose the skin, and then 0.5mL of the prepared RhB-VA-Col and RhB-VA-Col@Gen were applied on the 2 pieces of hair-removed skin. After 8 hours, the mice were anesthetized and killed, and the skins applied with RhB-VA-Col and RhB-VA-Col@Gen were taken, paraffin sections were prepared routinely, and fluorescence imaging was performed, such as Figure 3 shown.

[0079] 3. Results

[0080] from Figure 3 It can be concluded that the gentiopicroside composition of the present invention can significantly enter the skin for percutaneous absorption.

[0081] Application Example 2

[0082] 1. Construction of psoriasis skin model

[0083] This project used imiquimod (IMQ) to construct a mouse psoriasis skin model. IMQ was dissolved in an olive oil / acetone mixture (olive oil: acetone = 1:4).

[0084] Construction of psoriasis skin model: First, on day 0, the back skin of mice was prepared. On day 1, the back skin of mice was treated with 0.5% IMQ solution (100 μL); on day 4, the back skin of mice was treated with 0.5% IMQ solution (100 μL), and on week 2, the back skin of mice was treated with 0.2% IMQ solution (100 μL). After that, 0.2% IMQ solution was applied every other day for a total of 2 times to construct a mouse psoriasis model.

[0085] Administration: Administration started 24 hours after the second administration (5 days), blank group (i.e. Figure 4 Con) and the model group (i.e. Figure 4The IMQ in the sample was not treated, the concentration of the collagen derivative (Example 1) VA-Col monomer group was 0.2%, the concentration of the gentiopicroside Gen monomer group was 0.2%, and the concentration of the gentiopicroside composition (Example 7) VA-Col@Gen treatment group was 0.2%, and the positive control group was tacrolimus (TAC). Apply every day for a total of 7 days. Observe the skin lesions, take pictures, and analyze the repair effect of gentiopicroside VA-Col@Gen after the experiment.

[0086] 2. Results

[0087] from Figure 4 It can be seen that the gentiopicroside composition of the present invention has a significant effect on the treatment of psoriasis.

[0088] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. A gentiopicroside composition comprising a collagen derivative and gentiopicroside; in, The collagen derivative is a condensation product of compound A and collagen, wherein compound A is an effective ingredient having a carboxyl group.

2. The gentiopicroside composition according to claim 1, characterized in that The weight ratio of the collagen derivative to the gentiopicroside is 1:(0.01-10); and / or The collagen derivative and the gentiopicroside form nanoparticles.

3. The gentiopicroside composition according to claim 1 or 2, characterized in that The compound A is selected from retinoic acid or its derivatives, preferably retinoic acid and / or isotretinoin.

4. The gentiopicroside composition according to any one of claims 1 to 3, characterized in that The average molecular weight of the collagen derivative is 200-1000 KDa; and / or The average molecular weight of the collagen is 500 Da to 1,000,000 Da, preferably 500 Da to 300,000 Da, more preferably 500 Da to 5,000 Da, and even more preferably 500 to 1,500 Da.

5. The gentiopicroside composition according to any one of claims 1 to 4, characterized in that The preparation method of the collagen derivative comprises: subjecting collagen to a condensation reaction with a compound A to generate the collagen derivative.

6. The gentiopicroside composition according to claim 5, characterized in that The molar ratio of the collagen to the compound A is 1:(0.01-1); and / or, The condensation reaction uses EDC and NHS as condensation reagents; and / or, the molar ratio of the compound A to the EDC is 1:(0.5-1.5); and / or, the molar ratio of the compound A to the NHS is 1:(0.5-1.5); and / or, The condensation reaction time is 0.1 to 24 hours, preferably 0.1 to 4 hours, and / or the condensation reaction temperature is -5 to 10°C, preferably 0 to 4°C; and / or, Before the condensation reaction, the compound A is first dissolved in a mixed solution of PBS solution and tetrahydrofuran. Preferably, the pH of the PBS solution is 2-6, and / or the concentration of the PBS solution is 0.05-1M, and / or the volume ratio of the PBS solution to tetrahydrofuran is 1:(0.5-2).

7. A method for preparing a gentiopicroside composition, comprising: dissolving the collagen derivative in a solvent to obtain a solution A; Dissolving gentiopicroside in a solvent to obtain a solution B; Mix solution A and solution B.

8. The preparation method according to claim 7, characterized in that: The solvent is water; And / or, the mass concentration of the collagen derivative in the solution A is 0.001% to 50%; And / or, the mass concentration of gentiopicroside in the solution B is 0.001% to 50%.

9. The preparation method according to claim 7, characterized in that: The mixing conditions include: a stirring speed of 10 to 2000 rpm and a stirring time of 0.1 to 48 hours.

10. A skin care product comprising an effective ingredient and the gentiopicroside composition according to any one of claims 1 to 6 or the gentiopicroside composition obtained by the preparation method according to any one of claims 7 to 9.

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