Matrine-based ionic liquid conotoxin polypeptide solution, preparation, preparation method and application thereof

Abstract

The present invention discloses a matrine-based ionic liquid conotoxin polypeptide solution, a preparation method and application thereof, wherein the steps include: providing a conotoxin polypeptide mother liquor; adding matrine and an organic acid to water to obtain a matrine organic acid ion salt solution; mixing the matrine organic acid ion salt solution and the conotoxin polypeptide mother liquor to obtain a matrine-based ionic liquid conotoxin polypeptide solution. The prepared matrine-based ionic liquid conotoxin polypeptide solution has good stability and biocompatibility, and has a sustained-release and penetration-promoting effect, can be delivered through the skin, can avoid the degradation of polypeptide drugs in the acidic and enzymatic reaction conditions in the digestive system and the first-pass effect of the liver in clinical applications, and can reduce the influence of the skin barrier function when used externally.

Description

Technical Field

[0001] The present invention relates to the technical field of microemulsions, and in particular to a matrine-based ionic liquid conotoxin polypeptide solution, a preparation thereof, and a preparation method and application thereof. Background Art

[0002] Conotoxin peptides are complex mixtures of short peptides secreted by the venom glands lining the venom ducts and venom sacs of the cone snail (Conus geographus). They act as anesthetics for cone snails and have strong muscle penetration, making them potentially useful for reducing wrinkles and tightening skin. Furthermore, conotoxin peptides are antagonists of N and NMDA receptors. In particular, Con-G proteins are specific for the NMDA receptor 2B subtype. Therefore, conotoxin peptides are more selective than morphine in the treatment of various diseases and are non-addictive or have side effects within therapeutic doses. Furthermore, due to their association with NMDA receptors, they have potential in the treatment of Alzheimer's and Parkinson's diseases, as a means of alleviating drug withdrawal symptoms.

[0003] Oral administration remains the most preferred method due to its familiarity, ease of use, and high compliance. However, peptide-based drugs are susceptible to enzymatic degradation in the acidic environment of the stomach and are subject to a first-pass effect in the liver. Transdermal delivery can effectively avoid enzymatic degradation and hepatic metabolism of peptide preparations, but topical application is also affected by the skin barrier and is less effective.

[0004] Therefore, the existing technology still needs to be improved and developed. Summary of the Invention

[0005] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a matrine-based ionic liquid conotoxin polypeptide solution, preparation, and preparation method and application thereof, aiming to solve the problem that existing polypeptide component drugs are easily degraded by enzymatic reactions in the gastric acid environment when taken orally, and have a first-pass effect in the liver. When used externally, they are also affected by the skin barrier function and have poor effects.

[0006] The technical solutions of the present invention are as follows:

[0007] The first aspect of the present invention provides a method for preparing a matrine-based ionic liquid conotoxin polypeptide solution, comprising the steps of:

[0008] Providing cone snail toxin polypeptide mother solution;

[0009] adding matrine and an organic acid into water to obtain a matrine organic acid ion salt solution;

[0010] The matrine organic acid ion salt solution and the conotoxin polypeptide mother solution are mixed to obtain a matrine-based ionic liquid conotoxin polypeptide solution.

[0011] Optionally, the preparation steps of the cone snail toxin polypeptide mother solution include:

[0012] The conotoxin polypeptide is added into water to obtain a conotoxin polypeptide mother solution.

[0013] Optionally, the organic acid is selected from one or more of C6-C18 saturated fatty acids, unsaturated fatty acids with an unsaturation degree of 1-3, taurine, malic acid, and mandelic acid.

[0014] Optionally, the molar ratio of the matrine to the organic acid is 1:1 to 1:4.

[0015] The second aspect of the present invention provides a method for preparing a matrine-based ionic liquid conotoxin polypeptide preparation, which comprises the steps of:

[0016] mixing the emulsifier and the auxiliary agent to obtain a mixed emulsifier solution;

[0017] The matrine-based ionic liquid conotoxin polypeptide solution of the present invention is mixed with the mixed emulsifier solution to obtain a matrine-based ionic liquid conotoxin polypeptide preparation.

[0018] Optionally, the emulsifier is selected from one or more of polyacrylic acid, polyvinyl pyrrolidone, polyvinyl alcohol, and polyoxyethylene sorbitan fatty acid esters, and / or the auxiliary agent is selected from one or more of n-butanol, ethylene glycol, ethanol, propylene glycol, glycerol, polyglycerol fatty acid esters, and sorbitol fatty acid esters.

[0019] Optionally, the molar ratio of the emulsifier to the auxiliary agent is 1:0.1 to 1:10.

[0020] Optionally, the concentration of the conotoxin polypeptide in the matrine-based ionic liquid conotoxin polypeptide preparation is 0.0001 to 10 mg / mL.

[0021] The third aspect of the present invention provides a matrine-based ionic liquid conotoxin polypeptide preparation, wherein the preparation is prepared using the method for preparing the matrine-based ionic liquid conotoxin polypeptide preparation as described above.

[0022] In a fourth aspect, the present invention provides a method for preparing a matrine-based ionic liquid conotoxin polypeptide solution as described above, and uses the matrine-based ionic liquid conotoxin polypeptide solution obtained by the method for preparing the matrine-based ionic liquid conotoxin polypeptide solution as described above in the preparation of medicines or skin care products, or provides a method for preparing a matrine-based ionic liquid conotoxin polypeptide preparation as described above, and uses the matrine-based ionic liquid conotoxin polypeptide preparation obtained by the method for preparing the matrine-based ionic liquid conotoxin polypeptide preparation as described above in the preparation of medicines or skin care products, or provides a method for preparing a matrine-based ionic liquid conotoxin polypeptide preparation as described above in the preparation of medicines or skin care products.

[0023] Beneficial effects: The present invention provides a matrine-based ionic liquid conus toxin polypeptide solution, preparation, and preparation method and application thereof. The matrine-based ionic liquid conus toxin polypeptide solution prepared by the preparation method of the present invention has good stability and biocompatibility, has a sustained-release and penetration-promoting effect, and the conus toxin polypeptide therein has a high solubility, which can effectively improve the penetration of the conus toxin polypeptide and its absorption by the human body, and can be delivered transdermally, which can avoid the degradation of polypeptide drugs under acidic and enzymatic reaction conditions in the digestive system and the first-pass effect of the liver in clinical applications, and reduce the incidence of adverse reactions, and can reduce the impact of the skin barrier function when used externally. The matrine-based ionic liquid conus toxin polypeptide solution has a sustained-release effect, thereby reducing irritation to the skin during transdermal delivery, is mild and promotes transdermal absorption, and is green and safe. In addition, the matrine-based ionic liquid conus toxin polypeptide solution has firming, anti-wrinkle, and anti-aging effects, and has broad market prospects in various daily chemical products and preparation formulas. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a flow chart for preparing the matrine-based ionic liquid conotoxin polypeptide solution in an embodiment of the present invention.

[0025] Figure 2 This is a diagram showing the skin penetration effect of the blank control group in Example 1 of the present invention.

[0026] Figure 3 This is a diagram showing the skin penetration effect of the cone snail toxin polypeptide preparation in Comparative Example 1 of the present invention.

[0027] Figure 4 This is a diagram showing the skin penetration effect of the matrine-based ionic liquid conotoxin polypeptide preparation in Example 1 of the present invention.

[0028] Figure 5 These are the appearance test effect diagrams of the eye cream in Example 3 of the present invention, where (a) is the effect diagram before use of the eye cream, and (b) is the effect diagram after use of the eye cream. DETAILED DESCRIPTION

[0029] The present invention provides a matrine-based ionic liquid conotoxin polypeptide solution, preparation, and preparation method and application thereof. To make the objectives, technical solutions, and effects of the present invention clearer and more specific, the present invention is further described in detail below. It should be understood that the specific embodiments described herein are merely illustrative of the present invention and are not intended to limit the present invention.

[0030] The present invention provides a method for preparing a matrine-based ionic liquid conotoxin polypeptide solution, such as Figure 1 As shown, wherein the steps include:

[0031] S1. providing a cone snail toxin polypeptide stock solution;

[0032] S2, adding matrine and an organic acid into water to obtain a matrine organic acid ion salt solution;

[0033] S3. Mixing the matrine organic acid ion salt solution and the conotoxin polypeptide mother solution to obtain a matrine-based ionic liquid conotoxin polypeptide solution.

[0034] In the present embodiment, matrine is used as a cationic precursor and an organic acid is used as an anionic precursor, and matrine organic acid ion salt is synthesized by ionization salt-forming reaction. The matrine organic acid ion salt is a room temperature ionic liquid, which is a supramolecular material. Since the ionic liquid has a low vapor pressure, good stability, and special solubility for many inorganic salts and organic matter, the matrine organic acid ion salt solution can dissolve the conus toxin polypeptide well. At the same time, the matrine-based ionic liquid can wrap the conus toxin polypeptide to a certain extent to form micelles, enhance its stability, and promote absorption. The matrine organic acid ion salt solution and the conus toxin polypeptide mother liquor are mixed to obtain a matrine-based ionic liquid conus toxin polypeptide solution. The matrine-based ionic liquid conus toxin polypeptide preparation completely retains the main molecular skeleton of the matrine monomer, and therefore also gives the solution the effects of preventing skin psoriasis, anti-inflammatory, anti-allergic, promoting collagen production, and whitening. In addition, the matrine-based ionic liquid conotoxin polypeptide solution also retains the efficacy of organic acids. Organic acids have strong compatibility with epidermal tissues and can effectively open up ion channels between epidermal tissues. When passing through the skin, the solution can increase the fluidity of active ingredients such as conotoxin polypeptides and matrine in the epidermal tissues, thereby further increasing the penetration effect of active substances.

[0035] The present embodiment combines supramolecular material with conotoxin polypeptide, the matrine-based ionic liquid conotoxin polypeptide solution prepared has good stability and biocompatibility, there is the effect of slow-release facilitation, the matrine-based ionic liquid conotoxin polypeptide solution can effectively improve the penetration of conotoxin polypeptide and the absorption thereof by human body, oral or transdermal delivery can be carried out, it is possible to avoid the degradation of peptide drugs easily under acidic and enzymatic reaction conditions in the digestive system and the first-pass effect of the liver in clinical applications, and reduce the incidence rate of adverse reactions, the impact of skin barrier function when external use can be reduced. The solution has a slow-release effect so as to reduce the stimulation for skin when transdermal delivery, gentle and promote transdermal absorption, green and safe. In addition, the preparation has firming, anti-wrinkle, anti-aging effects, and has a broad market prospect in all kinds of daily chemical products and formulations.

[0036] In step S1, in one embodiment, the preparation step of the cone snail toxin polypeptide mother solution includes:

[0037] The conotoxin polypeptide is added into water to obtain a conotoxin polypeptide mother solution.

[0038] The conotoxin polypeptide is a complex mixture of short peptides secreted by the venom glands on the inner wall of the cone snail's venom duct and venom sac. It acts as an anesthetic for cone snails and has strong muscle penetration, making it potentially useful for reducing wrinkles and tightening skin. Conotoxin polypeptides are also antagonists of N receptors and NMDA receptors. In particular, Con-G protein is specific for the NMDA receptor 2B subtype. Therefore, conotoxin polypeptides are more selective than morphine in the treatment of various diseases and are non-addictive or have side effects within the therapeutic dose range. Furthermore, due to their association with NMDA receptors, they have potential in the treatment of Alzheimer's and Parkinson's diseases.

[0039] In step S2, matrine is used as a cationic precursor and an organic acid is used as an anionic precursor to synthesize a matrine organic acid ion salt through an ionization salt-forming reaction. The matrine organic acid ion salt is a room temperature ionic liquid. Since the ionic liquid has a low vapor pressure, good stability, and special solubility for many inorganic salts and organic substances, it can effectively increase the solubility of the active ingredient and increase the solubility and stability of the protein (for example, conotoxin polypeptide).

[0040] Matrine is an alkaloid extracted from Sophora flavescens, which has the effects of preventing psoriasis, combating inflammation, allergies, promoting collagen production, and whitening. Furthermore, matrine has antipyretic, analgesic, anticonvulsant, and neurostabilizing effects on the central nervous system; it has significant negative temporal and positive inotropic effects on the cardiovascular system, preventing and treating atherosclerosis and alleviating myocardial damage; and it has anti-liver damage and anti-fibrosis effects on the digestive system, as well as anti-tumor and anti-liver cancer effects.

[0041] In one embodiment, the organic acid is selected from one or more of C6-C18 saturated fatty acids, unsaturated fatty acids with an unsaturation degree of 1-3, taurine, malic acid, and mandelic acid, but is not limited thereto.

[0042] When the organic acid is selected from saturated or unsaturated fatty acids, it can increase the fat solubility and oil solubility of the matrine organic acid ion salt. This further enhances the compatibility of the matrine-based ionic liquid conotoxin polypeptide solution prepared from the matrine organic acid ion salt with epidermal tissue, effectively opening up ion channels between epidermal tissues, increasing the mobility of the active ingredient in the epidermal tissue, and thus enhancing the penetration effect of the active substance.

[0043] Taurine is a simple sulfur-containing amino acid in animals. It is odorless, slightly sour, and heat-stable. It exists in a bound form in human and animal bile bound to bile acid. It exists in a free form in tissues such as the brain, ovaries, heart, liver, breast milk, pineal gland, pituitary gland, retina, and adrenal glands, but does not participate in protein synthesis. Taurine is an essential amino acid for the human body and plays an important role in the development of the nervous system of fetuses and infants.

[0044] Malic acid, a major component of apples, is widely found in nature and is a widespread component of the human diet. Natural malic acid exhibits a variety of biological activities. In humans and other animals, it not only scavenges free radicals, provides antioxidant, anti-tumor, and antibacterial properties, but also suppresses obesity, improves mood, and promotes intestinal health. When malic acid is selected as the organic acid, it can promote the shedding of keratinocytes, increase skin collagen, and effectively improve dry skin.

[0045] In addition, the organic acid in this embodiment can also be selected from geranic acid.

[0046] In one embodiment, the molar ratio of matrine to the organic acid is 1:1 to 1:4. This ratio can better achieve ionization modification of matrine by the organic acid to generate a matrine organic acid ion salt.

[0047] In step S3, the step of mixing the matrine organic acid ion salt solution and the conotoxin polypeptide mother liquor to obtain a matrine-based ionic liquid conotoxin polypeptide solution specifically comprises:

[0048] Under stirring conditions, the matrine organic acid ion salt solution is added dropwise to the conotoxin polypeptide mother liquor and mixed to obtain a matrine-based ionic liquid conotoxin polypeptide solution. The matrine organic acid ion salt is a room temperature ionic liquid. Due to its low vapor pressure, good stability, and special solubility in many inorganic salts and organic substances, the matrine organic acid ion salt solution can well dissolve and encapsulate the conotoxin polypeptide and increase its stability.

[0049] The embodiment of the present invention also provides a matrine-based ionic liquid conus toxin polypeptide solution, wherein the solution is prepared by the method for preparing the matrine-based ionic liquid conus toxin polypeptide solution as described above in the embodiment of the present invention. The matrine-based ionic liquid conus toxin polypeptide solution comprises matrine organic acid ion salt, conus toxin polypeptide and water. The matrine-based ionic liquid conus toxin polypeptide solution is in a uniform and stable solution state. The matrine-based ionic liquid conus toxin polypeptide solution is mild and low in irritation, has good stability and biocompatibility, has a sustained-release and penetration-promoting effect, and can effectively improve the penetration of conus toxin polypeptide through the skin and its absorption by the human body. Moreover, the matrine-based ionic liquid conus toxin polypeptide solution retains the main molecular skeleton of the matrine monomer, thus giving the matrine-based ionic liquid conus toxin polypeptide solution the effects of preventing skin psoriasis, anti-inflammatory, anti-allergic, promoting collagen production, and whitening. Taking malic acid as an example, while retaining the characteristics of malic acid in promoting the shedding of keratinocytes, increasing skin collagen, and effectively improving dry skin, the matrine-based ionic liquid conotoxin polypeptide solution retains the biological activity and efficacy of matrine and organic acids.

[0050] The present invention also provides a method for preparing a matrine-based ionic liquid conotoxin polypeptide preparation, which comprises the steps of:

[0051] S4, mixing the emulsifier and the auxiliary agent to obtain a mixed emulsifier solution;

[0052] S5. Mixing the matrine-based ionic liquid conotoxin polypeptide solution described in the above embodiment of the present invention with the mixed emulsifier solution to obtain a matrine-based ionic liquid conotoxin polypeptide preparation.

[0053] In step S4, in one embodiment, the emulsifier is selected from one or more of polyacrylic acid, polyvinyl pyrrolidone, polyvinyl alcohol, and polyoxyethylene sorbitan fatty acid ester, but is not limited thereto.

[0054] In one embodiment, the auxiliary agent is selected from one or more of n-butanol, ethylene glycol, ethanol, propylene glycol, glycerol, polyglycerol fatty acid ester, and sorbitol fatty acid ester, but is not limited thereto.

[0055] In one embodiment, the molar ratio of the emulsifier to the auxiliary agent is 1:0.1 to 1:10. For example, the molar ratio of the emulsifier to the auxiliary agent can be 1:0.1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, etc.

[0056] In step S5, it should be noted that the emulsifier can encapsulate the conotoxin polypeptide to form micelles, further enhancing the stability of the conotoxin polypeptide, so that the conotoxin polypeptide can better penetrate into the skin. Therefore, the amount of emulsifier and adjuvant added can be adjusted according to the final permeability of the matrine-based ionic liquid conotoxin polypeptide preparation.

[0057] The matrine-based ionic liquid conotoxin polypeptide solution is mixed with the mixed emulsifier solution to obtain a matrine-based ionic liquid conotoxin polypeptide preparation. The matrine-based ionic liquid conotoxin polypeptide preparation is in a uniform and stable emulsion state.

[0058] In one embodiment, the matrine-based ionic liquid conotoxin polypeptide solution is mixed with the mixed emulsifier solution to obtain a matrine-based ionic liquid conotoxin polypeptide preparation, the steps specifically comprising:

[0059] Under stirring conditions, the matrine-based ionic liquid conotoxin polypeptide solution is added dropwise to the mixed emulsifier solution to obtain the matrine-based ionic liquid conotoxin polypeptide preparation.

[0060] In one embodiment, the concentration of the conotoxin polypeptide in the matrine-based ionic liquid conotoxin polypeptide preparation is 0.0001 to 10 mg / mL. At this concentration, the conotoxin polypeptide has better solubility and stability in the matrine-based ionic liquid conotoxin polypeptide preparation, while also achieving penetration and absorption on the skin surface.

[0061] The present invention also provides a matrine-based ionic liquid conotoxin polypeptide preparation, which is prepared using the method for preparing the matrine-based ionic liquid conotoxin polypeptide preparation described above in the present invention. The matrine-based ionic liquid conotoxin polypeptide preparation comprises a matrine organic acid ion salt, a conotoxin polypeptide, an emulsifier, an adjuvant, and water. The matrine-based ionic liquid conotoxin polypeptide preparation is mild and low-irritating, has improved stability and biocompatibility, and exhibits a sustained-release and penetration-promoting effect, effectively improving the permeation of the conotoxin polypeptide through the skin and its absorption by the human body. Furthermore, the matrine-based ionic liquid conotoxin polypeptide preparation retains the main molecular skeleton of the matrine monomer, thereby endowing the matrine-based ionic liquid conotoxin polypeptide preparation with the efficacy of preventing skin psoriasis, providing anti-inflammatory and anti-allergic effects, promoting collagen production, and whitening. Taking malic acid as an example, it also retains the characteristics of malic acid in promoting keratinocyte shedding, increasing skin collagen, and effectively improving dry skin. The matrine-based ionic liquid conotoxin polypeptide preparation simultaneously retains the biological activity and efficacy of matrine and organic acids.

[0062] The present invention also provides a use of the matrine-based ionic liquid conotoxin polypeptide solution prepared by the method for preparing the matrine-based ionic liquid conotoxin polypeptide solution according to the present invention in the preparation of medicines or skin care products.

[0063] The embodiment of the present invention further provides a use of the matrine-based ionic liquid conotoxin polypeptide solution described in the embodiment of the present invention in the preparation of medicines or skin care products.

[0064] The present invention also provides a use of the matrine-based ionic liquid conotoxin polypeptide preparation prepared by the method for preparing the matrine-based ionic liquid conotoxin polypeptide preparation according to the present invention in the preparation of medicines or skin care products.

[0065] The embodiment of the present invention also provides a use of the matrine-based ionic liquid conotoxin polypeptide preparation described in the embodiment of the present invention in the preparation of medicines or skin care products.

[0066] In a specific embodiment, the matrine-based ionic liquid conotoxin polypeptide preparation prepared by the method for preparing a matrine-based ionic liquid conotoxin polypeptide preparation described in the embodiments of the present invention, or the matrine-based ionic liquid conotoxin polypeptide preparation described in the embodiments of the present invention, is used to prepare an eye cream or face cream. In a more specific embodiment, the eye cream comprises a matrine-based ionic liquid conotoxin polypeptide preparation, wheat germ oil, sodium hyaluronate, vitamin B1, vitamin E, a surfactant, a cosurfactant, and water. The eye cream containing the matrine-based ionic liquid conotoxin polypeptide preparation can effectively promote the penetration of active ingredients, stimulate fibroblast regeneration and collagen production, and thus restore skin elasticity.

[0067] The matrine-based ionic liquid conotoxin polypeptide solution or preparation in the embodiment of the present invention can also be used in combination with other ingredients to prepare skin care products including but not limited to essences, eye mask essences, lotions, etc. to achieve different effects.

[0068] The present invention will be further described below with reference to specific examples.

[0069] Example 1

[0070] 0.01 mol of matrine and 0.01 mol of octanoic acid were added to a reaction vessel containing 20 mL of water and stirred for 12 hours to obtain a matrine organic acid ion salt solution. At room temperature, 0.001 g of conus toxin polypeptide was added to 10 mL of water and stirred to obtain a conus toxin polypeptide mother liquor. Under stirring conditions, the above-mentioned matrine organic acid ion salt solution was added dropwise to the conus toxin polypeptide mother liquor to obtain a matrine-based ionic liquid conus toxin polypeptide solution. At room temperature, the emulsifier polyvinyl pyrrolidone and the auxiliary agent sorbitol fatty acid ester were mixed in a molar ratio of 1:1 and thoroughly mixed to obtain a mixed emulsifier solution. Take 4 g of the mixed emulsifier solution and, under stirring conditions, add the above-mentioned matrine-based ionic liquid conus toxin polypeptide solution dropwise to 4 g of the mixed emulsifier solution. Observe the system for changes from turbidity to clarity or from clarity to turbidity to obtain a matrine-based ionic liquid conus toxin polypeptide preparation.

[0071] Example 2

[0072] 0.01 mol of matrine and 0.01 mol of malic acid were added to a reaction vessel containing 20 mL of water and stirred for 12 hours to obtain a matrine organic acid ion salt solution. At room temperature, 0.100 g of conus toxin polypeptide was added to 10 mL of water and stirred to obtain a conus toxin polypeptide mother liquor. Under stirring conditions, the above-mentioned matrine organic acid ion salt solution was added dropwise to the conus toxin polypeptide mother liquor to obtain a matrine-based ionic liquid conus toxin polypeptide solution. At room temperature, 0.848 g of emulsifier polyvinyl alcohol and auxiliary agent sorbitol fatty acid ester were mixed in a molar ratio of 1:1 and thoroughly mixed to obtain a mixed emulsifier solution. Take 4 g of the mixed emulsifier solution, under stirring conditions, add the above-mentioned matrine-based ionic liquid conus toxin polypeptide solution dropwise to 4 g of the mixed emulsifier solution, and observe the system from turbid to clear or from clear to turbid to obtain a matrine-based ionic liquid conus toxin polypeptide preparation.

[0073] Comparative Example 1

[0074] Preparation of Conus Toxin Peptide Preparation:

[0075] The conotoxin polypeptide preparation does not contain matrine, and its preparation method is the same as that of Example 1, except that matrine is not added.

[0076] The matrine-based ionic liquid conotoxin polypeptide preparation in Example 1 and the conotoxin polypeptide preparation in Comparative Example 1 were tested for transdermal delivery effects.

[0077] After the sample was applied to the epidermal model for 1 hour, the test results are shown in Table 1 and Figure 2-4 shown.

[0078] Table 1: Osmotic concentrations of different cone snail toxin peptide preparations

[0079]

[0080] As shown in Table 1, after the sample was applied to the epidermal model for 1 hour, the average penetration amount of the conotoxin polypeptide preparation group (Comparative Example 1) was 0.38 μg / mL, and the average penetration amount of the matrine-based ionic liquid conotoxin polypeptide preparation group (Example 1) was 0.71 μg / mL. Microscopic observation of the skin model revealed that the penetration depth of the conotoxin polypeptide in the matrine-based ionic liquid conotoxin polypeptide preparation group (Example 1) was significantly increased (e.g. Figure 4 and the penetration depth was significantly greater than that of the cone snail toxin polypeptide preparation group (Comparative Example 1, as shown in Figure 3 The results showed that the matrine-based ionic liquid conotoxin polypeptide preparation effectively opened up the ion channels between epidermal tissues, increased the fluidity and permeability of the conotoxin polypeptide in the epidermal tissues, and indicated an increase in the efficacy of the preparation.

[0081] Example 3

[0082] Preparation of anti-wrinkle and firming eye cream containing matrine-based ionic liquid conotoxin polypeptide preparation:

[0083] The following mass percentages were used to prepare the following: 0.1% of the matrine-based ionic liquid conotoxin polypeptide preparation in Example 1, 2.5% wheat germ oil, 0.5% sodium hyaluronate, 0.01% vitamin B1, 0.01% vitamin E, 2.0% surfactant, 0.5% cosurfactant, and the balance water to obtain an anti-wrinkle and firming eye cream containing the matrine-based ionic liquid conotoxin polypeptide preparation. The cream was then subjected to actual use tests, where it was applied to the eyes of subjects with wrinkles. The test results were as follows: Figure 5 As shown in (a) and (b), the eye cream has a significant wrinkle-reducing effect after 10 minutes of use. This suggests that the matrine-based ionic liquid conotoxin peptide preparation can effectively promote the penetration of active ingredients, stimulate fibroblast regeneration and collagen production, and thus restore skin elasticity.

[0084] In summary, the present invention provides a matrine-based ionic liquid conus toxin polypeptide solution, preparation and preparation method and application thereof, the matrine-based ionic liquid conus toxin polypeptide solution and preparation obtained by the preparation method of the present invention have good stability and biocompatibility, have the effect of sustained-release and enhanced penetration, the solubility of the conus toxin polypeptide is large, can effectively improve the penetration of the conus toxin polypeptide and the absorption thereof by the human body, can be delivered via a transdermal route, can avoid the degradation of polypeptide drugs under acidic and enzymatic reaction conditions in the digestive system and the first-pass effect of the liver in clinical applications, and reduce the incidence of adverse reactions, can reduce the impact of the skin barrier effect when used externally. The matrine-based ionic liquid conus toxin polypeptide solution and preparation have a sustained-release effect to reduce the irritation to the skin during transdermal delivery, are gentle and promote transdermal absorption, and are green and safe. In addition, the matrine-based ionic liquid conus toxin polypeptide solution and preparation have firming, anti-wrinkle, anti-aging effects, and have broad market prospects in various daily chemical products and preparation formulas.

[0085] It should be understood that the application of the present invention is not limited to the above examples. For those skilled in the art, improvements or changes can be made based on the above description. All these improvements and changes should fall within the scope of protection of the claims attached to the present invention.

Claims

1. A method for preparing a matrine-based ionic liquid conotoxin polypeptide preparation, characterized in that: Includes steps: mixing the emulsifier with the auxiliary agent to obtain a mixed emulsifier solution; The matrine-based ionic liquid conotoxin polypeptide solution is mixed with the mixed emulsifier solution to obtain a matrine-based ionic liquid conotoxin polypeptide preparation; the emulsifier is selected from one or more of polyacrylic acid, polyvinyl pyrrolidone, polyvinyl alcohol, and polyoxyethylene sorbitan fatty acid ester; the auxiliary agent is selected from one or more of n-butanol, ethylene glycol, ethanol, propylene glycol, glycerol, polyglycerol fatty acid ester, and sorbitol fatty acid ester; The concentration of the conotoxin polypeptide in the matrine-based ionic liquid conotoxin polypeptide preparation is 0.0001 to 10 mg / mL; The method for preparing the matrine-based ionic liquid conotoxin polypeptide solution comprises the following steps: Providing a conotoxin polypeptide stock solution; adding matrine and an organic acid into water to obtain a matrine organic acid ion salt solution; The matrine organic acid ion salt solution and the conotoxin polypeptide mother solution are mixed to obtain a matrine-based ionic liquid conotoxin polypeptide solution; the organic acid is selected from one or more of C6-C18 saturated fatty acids, unsaturated fatty acids with an unsaturation degree of 1-3, taurine, malic acid, and mandelic acid; The molar ratio of the emulsifier to the auxiliary agent is 1:0.1 to 1:

10.

2. The method for preparing the matrine-based ionic liquid conotoxin polypeptide preparation according to claim 1, characterized in that: The preparation steps of the conotoxin polypeptide mother solution include: The conotoxin polypeptide is added into water to obtain a conotoxin polypeptide mother solution.

3. The method for preparing the matrine-based ionic liquid conotoxin polypeptide preparation according to claim 1, characterized in that: The molar ratio of the matrine to the organic acid is 1:1 to 1:

4.

4. A matrine-based ionic liquid conotoxin polypeptide preparation, characterized in that: The preparation is obtained by the method for preparing the matrine-based ionic liquid conotoxin polypeptide preparation as described in any one of claims 1 to 3.

5. Use of a matrine-based ionic liquid conotoxin polypeptide preparation obtained by the preparation method of a matrine-based ionic liquid conotoxin polypeptide preparation as described in any one of claims 1 to 3 in the preparation of medicines or skin care products, or use of a matrine-based ionic liquid conotoxin polypeptide preparation as described in claim 4 in the preparation of medicines or skin care products.

6. The use according to claim 5, characterized in that: The skin care product is an eye cream, which includes the following ingredients in percentage by mass: 0.1% of matrine-based ionic liquid conotoxin polypeptide preparation, 2.5% of wheat germ oil, 0.5% of sodium hyaluronate, 0.01% of vitamin B1, 0.01% of vitamin E, 2.0% of surfactant and 0.5% of co-surfactant, and the balance is water.

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