High-solubility antibacterial and acne-removing berberine gel as well as preparation method and application thereof

Abstract

The invention discloses a high-solubility antibacterial and acne-removing berberine gel and a preparation method and application thereof.The preparation method comprises the steps that berberine and carbomer are used in a combined mode, the solubility of the berberine is changed by adjusting the pH value of a reaction system in the preparation process, more berberine is wrapped by means of the drug loading capacity of carbomer, and the anti-bacterial and acne-removing berberine gel is obtained. Therefore, the problems of poor solubility and low availability of berberine are improved. In addition, protease with positive electricity is added into a reaction system, positive electricity modification can be carried out on the surface of carbomer, bacteria with electronegative polysaccharide on the surface can be tended in a more targeted mode, and the antibacterial and acne-removing capacity is further enhanced. Meanwhile, the preparation method has the advantages of simple process, cheap and easily available raw materials, low production cost, simplicity in operation and control, and realization of large-scale industrial production.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and in particular to a highly soluble antibacterial and acne-removing berberine gel, and a preparation method and application thereof. Background Art

[0002] Acne, also known as pimples, is a common chronic inflammatory disease of the hair follicles and sebaceous glands. Its symptoms include facial lesions such as acne, papules, and pustules. The occurrence of acne is related to the following factors: excessive sebum secretion, blockage of the hair follicle sebaceous gland duct, bacterial infection and inflammatory response. When the skin secretes too much oil or the level of androgen in the human body increases, the sebaceous glands develop rapidly and secrete a large amount of sebum, which will clog the pores and easily induce acne. The keratinization of the hair follicle sebaceous gland duct is abnormal, and the sebum cannot be discharged normally, causing the skin to form keratin plugs and evolve into acne. The massive reproduction of Propionibacterium acnes in the hair follicles causes the sebum to produce free fatty acids, thereby aggravating the inflammatory cells and mediators, and finally inducing an inflammatory response. In view of the mechanism of acne occurrence, the acne removal effect can be achieved by controlling oil, softening the stratum corneum, and slowing down acne inflammation.

[0003] Coptis chinensis is the dried rhizome of the plant Coptis chinensis of the family Coptis chinensis. Because its rhizome is bead-shaped and yellow in color, it is called Coptis chinensis. It is a commonly used antibacterial and antiviral Chinese medicine in clinical practice. Coptis chinensis has a wide antibacterial spectrum and has a certain inhibitory effect on Gram-positive bacteria, Gram-negative bacteria and fungi. Its main component is isoquinolin alkaloids, among which berberine content is relatively high. Berberine, also known as berberine, is a yellow needle-shaped crystal. It is a quaternary ammonium alkaloid isolated from the traditional Chinese medicine Coptis chinensis. It is the main active ingredient of Coptis chinensis antibacterial and has good antibacterial effect. Berberine is a multi-target compound that can act on multiple pathways of bacterial metabolism and multiple processes of bacterial pathogenicity. Therefore, berberine has good application prospects in acne removal.

[0004] Although berberine has strong antibacterial activity, due to its structure belonging to quaternary ammonium base, it has poor water solubility and low bioavailability in plasma. The amount of oral berberine absorbed is small, and a large amount of berberine accumulates in the intestine and interacts with intestinal bacteria. In addition, berberine is converted into easily absorbed metabolites such as dihydroberberine by nitroreductase in intestinal bacteria. The antibacterial effect of these metabolites is weaker, which may represent one of the mechanisms of bacterial self-protection. Berberine is an important lead compound with great optimization potential in drug development, but due to its extremely low water solubility, poor oral absorption, and poor oral bioavailability, it has become the main factor limiting the clinical application of berberine. Free berberine can be slowly dissolved in water, easily soluble in hot water or hot ethanol, and not very soluble in cold ethanol. The hydrochloride of berberine has a low solubility in water, is more soluble in boiling water, and is difficult to dissolve in ethanol. When berberine combines with large molecular organic acids, such as glycyrrhizic acid, baicalin, and rhubarb tannin, the salts formed have very low solubility in water.

[0005] Therefore, it is of positive significance to improve the problems of poor solubility and low utilization of berberine and apply it to the preparation of acne-removing products.

[0006] In view of this, the present invention is proposed. Summary of the invention

[0007] The purpose of the present invention is to provide a highly soluble antibacterial acne-removing berberine gel and a preparation method and application thereof. The berberine gel obtained by the preparation method of the present invention can carry more berberine, thereby better exerting the acne-removing ability of berberine.

[0008] The present invention is achieved in that:

[0009] In a first aspect, the present invention provides a highly soluble antibacterial and acne-removing berberine gel, wherein the berberine gel is a nanoparticle structure, which comprises, from the inside to the outside, berberine, carbomer encapsulating berberine, and a protease modified on the surface of the carbomer.

[0010] In a second aspect, the present invention provides a method for preparing the above-mentioned antibacterial and anti-acne berberine gel, which comprises: adjusting the pH of the reaction system, and adding positively charged protease to the reaction system, and obtaining the berberine gel after it is fully dissolved.

[0011] In some embodiments, the preparation method includes: adjusting the pH of the carbomer solution to 7.5-10, then adding berberine thereto to form a mixed solution, and adjusting the pH of the mixed solution to 3.0-6.0, stirring evenly, and then adding positively charged protease and carbomer dry powder in sequence, and obtaining berberine gel after sufficient dissolution.

[0012] In some embodiments, the preparation method comprises: adjusting the pH of the carbomer solution to 7.5-10, and adjusting the pH of the berberine solution to 3.0-6.0, and then mixing the pH-adjusted carbomer solution and berberine solution, stirring evenly, adding positively charged protease, and obtaining berberine gel after sufficient dissolution.

[0013] In some embodiments, the preparation method further comprises: adjusting the pH of the solution to which the protease is added to 3.0-7.0 after the protease is added.

[0014] In some embodiments, the reaction conditions for preparing the mixed solution and berberine solution are: heating and stirring at 40-70°C.

[0015] In some embodiments, the conditions for sufficient dissolution are: constant temperature heating and stirring at 40-70° C. and 1400-2000 rpm.

[0016] In some embodiments, the carbomer in the carbomer solution and the carbomer dry powder in the above preparation method is selected from at least one of carbomer 934, carbomer 940, carbomer 941, carbomer 974P and carbomer 980; the positively charged protease includes lysozyme; and berberine includes berberine hydrochloride.

[0017] In some embodiments, the carbomer is carbomer 940.

[0018] In some embodiments, the mass ratio of the above-mentioned carbomer 940 to berberine and lysozyme is: 5-10:1:2.

[0019] In a third aspect, the present invention also provides the use of the berberine gel in the preparation of medicines or cosmetics with antibacterial and anti-acne effects.

[0020] The present invention has the following beneficial effects:

[0021] The present invention uses berberine in combination with carbomer, and changes the solubility of berberine by adjusting the pH of the reaction system during the preparation process, and uses the drug-carrying capacity of carbomer to encapsulate more berberine, thereby improving the problems of poor solubility and low utilization of berberine. In addition, the present invention also adds a positively charged protease in the reaction system, which can be positively modified on the surface of carbomer, and can be more targeted to bacteria with negatively charged polysaccharides on the surface, further enhancing the antibacterial and acne-removing ability. At the same time, the preparation method of the present invention has a simple process, cheap and easy-to-obtain raw materials, low production cost, simple operation and control, and can be mass-produced industrially. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0023] Figure 1 This is the administration of Staphylococcus aureus in Experimental Example 1;

[0024] Figure 2 This is the administration of Klebsiella pneumoniae in Experimental Example 1. DETAILED DESCRIPTION

[0025] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0026] Carbomer is the most commonly used matrix, which has a certain acidity and can be neutralized by alkaline substances, such as sodium hydroxide, triethanolamine, etc. It has good thixotropy and is mainly used as a thickener, emulsifier and suspending agent in pharmaceutical preparations. It can improve the stability and solubility of the drug and is conducive to the transdermal absorption of the drug. At the same time, carbomer is also a topical drug that can be made into a gel as a drug carrier medium. It usually has anti-inflammatory, wound healing, antioxidant, angiogenesis, skin repair and other effects. In order to improve the problem of poor solubility and low utilization of berberine, the inventors of the present invention used the physicochemical properties of berberine, used carbomer as a carrier, and changed its pH to make a berberine gel to improve the bioavailability and therapeutic effect of berberine. The first preparation method of the above-mentioned berberine gel is carried out as follows:

[0027] S1. Add carbomer dry powder to distilled water, stir and adjust the pH to 7.5-10, and leave overnight to fully swell to form a carbomer matrix solution.

[0028] In the present invention, carbomer is used as a drug carrier for berberine, which may be carbomer 934, carbomer 940, carbomer 941, carbomer 974P and carbomer 980, or other types of carbomer. Preferably, carbomer 940 is selected as the carbomer.

[0029] Here, the pH is adjusted to alkaline, and the pH can be adjusted to 7.5, 8.0, 8.5, 9.0, 9.5 or 10.0, or any other value between 7.5 and 10. The purpose of adjusting the pH of the solution to alkaline is to fully disperse the carbomer in the distilled water and to be evenly distributed.

[0030] S2. Add berberine to hot water and adjust the pH to 3-6, mix thoroughly, then add the swollen carbomer base solution in S1, and stir to mix.

[0031] The pH of the distilled water to which berberine is added is adjusted to be acidic, and the pH can be adjusted to 3, 3.5, 4, 4.5, 5, 5.5 or 6, or any other value between 3 and 6. The purpose of adjusting the pH of the solution to be acidic is to form an acidic environment, so that the carbomer in the carbomer matrix solution added later can form a structure capable of encapsulating berberine.

[0032] In the present invention, berberine can be berberine hydrochloride or other types of berberine.

[0033] The temperature of the hot water for dissolving berberine is 40-70° C. Specifically, the temperature of the hot water can be 40° C., 50° C., 60° C. or 70° C., or any value between 40-70° C. The solubility of berberine is increased by dissolving berberine in hot water.

[0034] S3. Add positively charged protease to the mixed solution obtained in S2, and obtain berberine gel after it is fully dissolved.

[0035] In this method, the addition of positively charged protease on the basis of forming a core-shell structure is also to modify the surface of the carbomer coated with berberine, so as to improve the antibacterial ability of the gel product.

[0036] On this basis, the role of adding a positively charged protease is to couple the protease to the surface of the carbomer. Through the modification of the protease, a nanoparticle is obtained, and the nanoparticle is composed of berberine, carbomer encapsulating berberine, and protease modified on the surface of the carbomer from the inside to the outside. The positive charge modification on the surface of the carbomer is because most of the bacteria that cause symptoms such as inflammation and acne are polysaccharides with negative charges on the surface. The nanoparticles of the present invention can enhance the penetration through positive charge modification, thereby improving the antibacterial ability.

[0037] Among them, the positively charged protease may be lysozyme.

[0038] The condition for sufficient dissolution is constant temperature heating and stirring at 40-70°C and 1400-2000rpm. Specifically, the heating temperature can be 40°C, 50°C, 60°C or 70°C, or any value between 40-70°C; the stirring speed can be 1400rpm, 1500rpm, 1600rpm, 1700rpm, 1800rpm, 1900rpm or 2000rpm, or any value between 1400-2000rpm.

[0039] Optionally, after adding the protease in S3, the pH of the reaction system can be adjusted according to the pH requirements of the subsequent product. Specifically, the pH of the reaction solution after adding lysozyme is adjusted to 3-7. The pH can be adjusted to 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5 or 7, or any other value between 3-7.

[0040] In the preparation method, the mass ratio of carbomer to berberine and protease is 5:1:2.

[0041] The second preparation method of the berberine gel of the present invention is carried out according to the following steps:

[0042] S1. Add carbomer dry powder into distilled water and stir until completely dissolved to prepare a carbomer solution.

[0043] S2. Adjust the pH of the carbomer solution to 7.5-10, add berberine, heat and stir at a constant temperature to form a mixed solution.

[0044] Before adding berberine, the pH of the carbomer solution is adjusted to alkaline, and the pH can be adjusted to 7.5, 8.0, 8.5, 9.0, 9.5 or 10.0, or any other value between 7.5 and 10. The purpose of adjusting the pH of the carbomer solution to alkaline is to promote the formation of a state in which two solutes are evenly dispersed in the mixed solution after berberine is subsequently added to the carbomer solution.

[0045] The constant temperature heating and stirring is specifically as follows: the carbomer solution with berberine added is stirred at 40-70° C. The heating temperature may be 40° C., 50° C., 60° C. or 70° C., or any value between 40-70° C.

[0046] S3. Adjust the pH of the mixed solution to 3-6, then add positively charged protease and stir evenly.

[0047] After adding berberine, the pH of the mixed solution is adjusted to acidic, and the pH can be adjusted to 3, 3.5, 4, 4.5, 5, 5.5 or 6, or any other value between 3 and 6. The purpose of adjusting the pH of the mixed solution to acidic is to allow carbomer to wrap berberine in an acidic environment to form a core-shell structure.

[0048] On this basis, the role of adding a positively charged protease is to couple the protease to the surface of the carbomer. Through the modification of the protease, a nanoparticle is obtained, and the nanoparticle is composed of berberine, carbomer encapsulating berberine, and protease modified on the surface of the carbomer from the inside to the outside. The positive charge modification on the surface of the carbomer is because most of the bacteria that cause symptoms such as inflammation and acne are polysaccharides with negative charges on the surface. The nanoparticles of the present invention can enhance the penetration through positive charge modification, thereby improving the antibacterial ability.

[0049] Among them, the positively charged protease may be lysozyme.

[0050] S4. Add carbomer dry powder again, heat and stir at a constant temperature, and obtain a gel after it is completely dissolved.

[0051] It should be noted that, compared with the first preparation method, the purpose of adding carbomer dry powder again in this method is to form a gel environment for the nanoparticles obtained in the above process, so that the gel solution becomes more viscous and has a better retention effect; while the addition of carbomer again in S4 does not affect the formation of berberine gel nanostructures.

[0052] Constant temperature heating and stirring are carried out under the conditions of 40-70°C and 1400-2000rpm. Specifically, the heating temperature can be 40°C, 50°C, 60°C or 70°C, or any value between 40-70°C; the stirring speed can be 1400rpm, 1500rpm, 1600rpm, 1700rpm, 1800rpm, 1900rpm or 2000rpm, or any value between 1400-2000rpm.

[0053] In the preparation method, the mass ratio of carbomer to berberine and protease is 5-10:1:2. The mass of carbomer here includes the total mass of carbomer dry powder in the carbomer solution in S1 and the carbomer dry powder in S4.

[0054] Optionally, before adding carbomer dry powder in S4, the pH of the reaction system can be adjusted according to the requirements of the pH of the subsequent product. Specifically, the pH of the reaction solution after adding lysozyme is adjusted to 3-7. The pH can be adjusted to 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5 or 7, or any other value between 3-7.

[0055] The inventors have proved through experiments that the berberine gel prepared by the above two methods can play the role of anti-inflammatory, wound healing, anti-oxidation, angiogenesis, skin repair, etc. when carbomer gel and berberine are used separately, and berberine can play the role of antibacterial, anti-inflammatory, anti-tumor, etc.; and when carbomer gel and berberine are used together, the effect of pharmacodynamic interaction can be achieved, including:

[0056] 1. Promote drug penetration and absorption: The permeation-promoting effect of carbomer gel can improve the permeability and absorption of berberine, thereby enhancing its efficacy.

[0057] 2. Enhance the efficacy of drugs: Berberine can enhance the efficacy of carbomer gel, making its anti-inflammatory and analgesic effects more significant.

[0058] 3. Reduce adverse reactions of drugs: Berberine can alleviate the adverse reactions that may be caused by carbomer gel, such as skin irritation, allergies, etc.

[0059] 4. Delay drug release and maintain drug stability: Carbomer gel can be used as a carrier to delay the release of berberine, and it can also improve its stability so that it can maintain better efficacy during storage and use.

[0060] In summary, the berberine gel provided by the present invention can increase the amount of berberine encapsulated in carbomer due to the control of the pH of the reaction system, and the synergistic effect of berberine and carbomer can improve the antibacterial effect, and finally the antibacterial ability is further improved under the action of positively charged proteases.

[0061] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0062] Example 1

[0063] Place 0.50g of Carbomer 940 dry powder in a beaker, measure 100mL of distilled water, add an equal amount of distilled water in batches, 25mL / time, and stir while adding until the dry powder is completely dissolved. In the solution, adjust the pH to 7.5 with c=1.0008mol / LNaOH standard solution, then add 0.10g of berberine hydrochloride, and stir for 1h at 40℃ in a constant temperature heating stirrer. Then adjust the solution pH to 5.5 with c=1.002mol / L hydrochloric acid standard solution, weigh 0.20g of lysozyme, add it to the solution and stir for 30min. Adjust the pH of the solution to 7.0 again with standard NaOH solution, add 0.50g of Carbomer 940 dry powder again, and heat and stir at 40℃ and 1400rpm to completely dissolve it to obtain a gel.

[0064] Example 2

[0065] Place 0.50g of Carbomer 940 dry powder in a beaker, measure 100mL of distilled water, add an equal amount of distilled water in batches, 25mL / time, and stir while adding until the dry powder is completely dissolved. In the solution, adjust the pH to 8.0 with c=1.0008mol / LNaOH standard solution, then add 0.10g of berberine hydrochloride, and stir for 1h at 50℃ in a constant temperature heating stirrer. Then adjust the solution pH to 6 with c=1.002mol / L hydrochloric acid standard solution, weigh 0.20g of lysozyme, add it to the solution and stir for 30min. Adjust the pH of the solution to 7.0 again with standard NaOH solution, add 0.50g of Carbomer 940 dry powder again, and heat and stir at 50℃ and 1600rpm to completely dissolve it to obtain a gel.

[0066] Example 3

[0067] Place 0.50g of Carbomer 940 dry powder in a beaker, measure 100mL of distilled water, add an equal amount of distilled water in batches, 25mL / time, and stir while adding until the dry powder is completely dissolved. In the solution, adjust the pH to 7.5 with c=1.0008mol / LNaOH standard solution, then add 0.10g of berberine hydrochloride, and stir at 60℃ for 1h in a constant temperature heating stirrer. Adjust the solution pH to 5.5 with c=1.002mol / L hydrochloric acid standard solution, weigh 0.20g of lysozyme, add it to the solution and stir for 30min. Adjust the pH of the solution to 7.0 again with standard NaOH solution, add 0.50g of Carbomer 940 dry powder again, and heat and stir at 60℃ and 1800rpm to completely dissolve it to obtain a gel.

[0068] Example 4

[0069] Sprinkle 0.50g of Carbomer 940 dry powder evenly in 100mL of distilled water, stir continuously, adjust pH=8.0, make it fully dispersed and evenly dispersed, leave overnight, and fully swell. Dissolve 0.10g of berberine hydrochloride in 50℃ distilled water, and adjust pH=5.5. After fully mixing, add it to the swollen Carbomer matrix solution, mix evenly, and stir continuously. Weigh 0.20g of lysozyme, add it to the solution and stir for 30min. At the same time, measure its pH value, adjust the pH value to 7.0, and heat and stir at a constant temperature of 50℃ and 1600rpm to completely dissolve it to obtain a gel.

[0070] Example 5

[0071] Sprinkle 0.50g of Carbomer 940 dry powder evenly in 100mL of distilled water, stir continuously, adjust pH to 8.5, make it fully dispersed, leave overnight, and fully swell. Dissolve 0.10g of berberine hydrochloride in 60℃ distilled water, and adjust pH to 6.0. After fully mixing, add it to the swollen Carbomer matrix solution, mix evenly, and stir continuously. Weigh 0.20g of lysozyme, add it to the solution and stir for 30min. At the same time, measure its pH value, adjust the pH value to 7.0, and heat and stir at a constant temperature of 60℃ and 1800rpm to completely dissolve it to obtain a gel.

[0072] Example 6

[0073] Sprinkle 0.50g of Carbomer 940 dry powder evenly in 100mL of distilled water, stir continuously, adjust pH=8.0, make it fully dispersed and evenly dispersed, leave overnight to fully swell. Dissolve 0.10g of berberine hydrochloride in 70℃ distilled water, adjust pH=6.5, and after fully mixing, add it to the swollen Carbomer matrix solution, mix evenly, and stir continuously. Weigh 0.20g of lysozyme, add it to the solution and stir for 30min. At the same time, measure its pH value, adjust pH value to 7.0, heat and stir at constant temperature at 70℃ and 2000rpm to make it completely dissolved to obtain gel.

[0074] Comparative Example 1

[0075] Place 0.50g of Carbomer 940 dry powder in a beaker, measure 100mL of distilled water, add an equal amount of distilled water in batches, 25mL / time, and stir while adding until the dry powder is completely dissolved. In the solution, measure the original pH value, pH = 3.5, add 0.10g of berberine hydrochloride, and stir at 40℃ for 1h in a constant temperature heating stirrer. Weigh 0.20g of lysozyme, add it to the solution and stir for 30min. Add 0.50g of Carbomer 940 dry powder again, heat and stir at 40℃ and 1400rpm to completely dissolve it and obtain a gel.

[0076] Experimental Example 1

[0077] This experimental example is a comparison of the antibacterial effects of the products prepared in the embodiment and the comparative example. The experimental method is as follows:

[0078] Staphylococcus aureus and Klebsiella pneumoniae were incubated with shaking in Luria–Bertani (LB) or Mueller Hinton medium at 37°C overnight. Subsequently, the bacteria were gently removed from the agar plate using a sterilized loop and then incubated in the medium at 37°C overnight. In the next step, 0.5 mL of cultured Staphylococcus aureus and Klebsiella pneumoniae were taken and incubated with the sample or LB medium in distilled water. After that, the above system was incubated at 37°C for 24 hours. The antibacterial performance was determined by counting colony forming units. Each sample was then cultured on an agar plate for 12 hours to measure the number of bacterial colonies. To test the antibacterial properties of the membrane, we placed the membrane on liquid LB agar and allowed the agar to solidify naturally. The membrane remained naturally on the agar surface, and the diameter of the membrane used for the antibacterial experiment was 6 mm. 1 mL of cultured Staphylococcus aureus and Klebsiella pneumoniae culture (1×10 7 After the whole culture was incubated overnight, the colonies were counted.

[0079] The concentrations of Staphylococcus aureus and Klebsiella pneumoniae when coating the plates are shown in Table 1:

[0080] Table 1 Concentrations of Staphylococcus aureus and Klebsiella pneumoniae

[0081]

[0082] The antibacterial effects of the embodiments and comparative examples are shown in the figure. Figure 1 The administration of Staphylococcus aureus in Example 1 is shown in FIG. Figure 2 This is the administration of Klebsiella pneumoniae in Example 1.

[0083] Experimental Example 2

[0084] The following is an explanation of a typical application example of applying the gel obtained in Example 1, as follows:

[0085] A total of 100 patients with acne, acne scars, blackheads and pustular acne were selected, aged 18 to 35 years old, including 28 patients with acne, including 20 females and 8 males; 36 patients with pustular acne, including 28 females and 8 males; 18 patients with blackheads, including 12 females and 6 males; 18 patients with acne scars, including 14 females and 4 males.

[0086] The following treatment plan was adopted for these 100 patients: the antibacterial acne gel of the present invention was used once a day, and the antibacterial acne gel obtained in Example 1 of the present invention was applied to the cleaned affected area at night each time, and massaged until absorbed, and then washed off the next morning. A course of treatment was 20 days, and the medication was used continuously for 20 days, and a follow-up visit was conducted once a week.

[0087] The efficacy evaluation criteria include: (1) Cure: clinical symptoms completely disappear, pain and discomfort completely disappear, and normal life is restored; (2) Effective: all or major symptoms are significantly eliminated, pain and discomfort obviously disappear, but mild symptoms still occur, and normal life is only partially restored; (3) Ineffective: no obvious change in symptoms.

[0088] The results of clinical application experiments are shown in Table 2:

[0089] Table 2 Clinical application effect of Example 1

[0090] cure efficient invalid Efficiency Acne 20 8 0 100％ Pustular pox 28 8 0 100％ acne 12 6 0 100％ Acne scars 14 4 0 100％

[0091] It can be seen from the application experimental results in the above table that the anti-acne gel of the present invention has a significant therapeutic effect on skin diseases such as acne, blackheads and pustular acne.

[0092] The typical application example of applying the gel obtained in Comparative Example 1 is further described as follows:

[0093] A total of 100 patients with acne, acne marks, blackheads and pustular acne were selected, aged 18 to 35 years old, including 28 acne patients, including 20 females and 8 males; 36 pustular acne patients, including 28 females and 8 males; 18 blackhead patients, including 12 females and 6 males; 18 blackhead patients, including 14 females and 4 males. The following treatment plan was adopted for these 100 patients: the antibacterial acne gel of the present invention was used once a day, and the antibacterial acne gel obtained in Example 1 of the present invention was applied to the affected area after cleaning at night each time, and massaged until absorbed, and then washed off the next morning, with 20 days as a course of treatment, continuous medication for 20 days, and a follow-up visit once a week.

[0094] The efficacy evaluation criteria include: (1) Cure: clinical symptoms completely disappear, pain and discomfort completely disappear, and normal life is restored; (2) Effective: all or major symptoms are significantly eliminated, pain and discomfort obviously disappear, but mild symptoms still occur, and normal life is only partially restored; (3) Ineffective: no obvious change in symptoms.

[0095] The results of clinical application experiments are shown in Table 3:

[0096] Table 3 Clinical application effect of comparative example 1

[0097] cure efficient invalid Efficiency Acne 0 7 21 25.0％ Pustular pox 0 10 26 27.8％ acne 0 6 12 33.3％ Acne scars 0 4 14 22.2％

[0098] From the experimental results in the above table, it can be seen that the anti-acne gel in Comparative Example 1 of the present invention has poor efficacy on skin diseases such as acne, blackheads, acne marks and pustular acne. Therefore, it can be proved that the berberine gel obtained by adjusting the pH during the reaction process of the present invention can reduce the amount of berberine coated in carbomer, thereby improving the antibacterial effect.

[0099] In order to further illustrate the efficacy of the anti-acne gel of the present invention, a large number of clinical comparative experiments were conducted in this experimental example. The following is an example of the commonly used retinoic acid cream, as follows:

[0100] A total of 100 patients with acne, acne marks, blackheads and pustular acne were selected, aged 16 to 38 years old, including 35 patients with acne, including 26 females and 9 males; 28 patients with acne, including 20 females and 8 males; 25 patients with pustular acne, including 7 females and 18 males; 12 patients with acne marks, including 6 females and 6 males.

[0101] The following treatment plan was adopted for these 100 patients: use retinoic acid cream once a day, apply it to the affected area after washing at night, massage until absorbed, and then wash it off the next morning. One course of treatment is 20 days, and the medication should be used continuously for 20 days, and the patient should be followed up once a week.

[0102] The efficacy evaluation criteria include: (1) Cure: clinical symptoms completely disappear, pain and discomfort completely disappear, and normal life is restored; (2) Effective: all or major symptoms are significantly eliminated, pain and discomfort obviously disappear, but mild symptoms still occur, and normal life is only partially restored; (3) Ineffective: no obvious change in symptoms.

[0103] The experimental results are shown in Table 4:

[0104] Table 4 Clinical application effect of the comparison group

[0105] cure efficient invalid Efficiency Acne 7 20 8 77.1％ Pustular pox 9 12 4 84.0％ acne 15 4 9 67.9％ Acne scars 2 4 6 50％

[0106] Comparing the experimental results of the above-mentioned tretinoin cream with the clinical application experimental results of the anti-acne gel of the present invention, it can be seen that the anti-acne cream provided by the present invention has a more thorough and significant therapeutic effect.

[0107] In general, it can be seen from the above application experiment results and comparative experiment results that the anti-acne cream of the present invention has a significant therapeutic effect on skin diseases such as acne, blackheads and pustular acne.

[0108] Experimental Example 3

[0109] This experimental example is a typical clinical case to demonstrate the acne-removing effect of the berberine gel of the present invention, as follows:

[0110] Clinical application example 1:

[0111] Ms. Huang, age: 22 years old, skin type: mixed dry in summer, dry skin in winter, experience of using the highly soluble antibacterial acne-removing berberine gel of Example 1 of the present invention: mature acne can be completely deflated after two days of use, and it has no effect on acne without pus. Skin feel: the face dries quickly after application, and the face feels tight.

[0112] Clinical application example 2:

[0113] Mr. Li, age: 32 years old, skin type: mixed dry skin, symptoms: allergic acne, feeling of using the highly soluble antibacterial acne-removing berberine gel of Example 2 of the present invention: for acne with pus, apply it three times a day, and it can be completely deflated after three consecutive days of use. Skin feeling: the face will feel very tight after application.

[0114] Clinical application example 3:

[0115] Ms. Zhang, 30 years old, had symptoms of closed comedones. She applied the highly soluble antibacterial acne-removing berberine gel of Example 3 of the present invention twice a day. After 7 consecutive days of use, the acne disappeared.

[0116] Clinical application example 4:

[0117] Mr. Lin, 25 years old, had symptoms of papules. He applied the highly soluble antibacterial acne-removing berberine gel of Example 4 of the present invention twice a day. After continuous use for 10 days, the papules disappeared.

[0118] Clinical application example 5:

[0119] Mr. Li, 28 years old, had symptoms of pustular acne. He used the highly soluble antibacterial acne-removing berberine gel of Example 5 of the present invention and applied it three times a day. After continuous use for 15 days, the symptoms of pustular acne were alleviated, leaving only a few acne marks.

[0120] Clinical application example 6:

[0121] Mr. Zhang, 18 years old, had symptoms of open acne. He used the highly soluble antibacterial acne-removing berberine gel of Example 6 of the present invention, applied it twice a day. After continuous use for 10 days, the acne disappeared and the melanin faded.

[0122] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A highly soluble antibacterial acne-removing berberine gel, characterized in that: The berberine gel is a nanoparticle structure, which comprises berberine, carbomer encapsulating berberine, and positively charged protease modified on the surface of carbomer from the inside to the outside.

2. The method for preparing the highly soluble antibacterial acne-removing berberine gel according to claim 1, characterized in that: include: The pH of the reaction system is adjusted, and positively charged protease is added to the reaction system, and berberine gel is obtained after it is fully dissolved.

3. The preparation method according to claim 2, characterized in that: The pH of the carbomer solution is adjusted to 7.5-10, and then berberine is added thereto to form a mixed solution, and the pH of the mixed solution is adjusted to 3.0-6.

0. After stirring evenly, positively charged protease and carbomer dry powder are added in sequence, and berberine gel is obtained after it is fully dissolved.

4. The preparation method according to claim 2, characterized in that: The pH of the carbomer solution is adjusted to 7.5-10, and the pH of the berberine solution is adjusted to 3.0-6.0, and then the carbomer solution and berberine solution after the pH adjustment are mixed, and after stirring evenly, positively charged protease is added, and berberine gel is obtained after it is fully dissolved.

5. The preparation method according to claim 3 or 4, characterized in that: The preparation method further comprises: adjusting the pH of the solution to which the protease is added to 3.0-7.0 after the protease is added.

6. The preparation method according to claim 3 or 4, characterized in that: The reaction conditions for preparing the mixed solution and the berberine solution are: heating and stirring at 40-70°C.

7. The preparation method according to claim 3 or 4, characterized in that: The conditions for sufficient dissolution are: constant temperature heating and stirring at 40-70°C and 1400-2000 rpm.

8. The preparation method according to any one of claims 5 to 7, characterized in that: The carbomer in the carbomer solution and the carbomer dry powder in the preparation method is selected from at least one of carbomer 934, carbomer 940, carbomer 941, carbomer 974P and carbomer 980; the positively charged protease includes lysozyme; the berberine includes berberine hydrochloride; Preferably, the carbomer is carbomer 940.

9. The preparation method according to claim 8, characterized in that: The mass ratio of the carbomer 940 to berberine and lysozyme is 5-10:1:

2.

10. Use of the berberine gel according to claim 1 in the preparation of antibacterial and anti-acne medicines or cosmetics.

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