Prescription and preparation method of medical antipyretic and antipruritic ointment

Abstract

The application belongs to the technical field of medical preparations, and discloses a medical heat-reducing and itching-relieving cream and a preparation method thereof. The cream is composed of functional active ingredients and matrix adjuvants, and the core components include glycyrrhizic acid modified beta-glucan, organic selenium grafted sodium hyaluronate, hydroxypropyltrimethylammonium chloride chitosan loaded curcumin compound, triple immunoadjuvant and peppermint oil. The preparation method includes the preparation of water phase and oil phase, emulsification homogenization and low-temperature active compound steps. Through the synergistic compatibility of modified polysaccharides, nano drug-loaded compounds and immunoadjuvants, combined with the high-pressure homogenization and low-temperature compound process, the cream prepared is fine and stable. The product has the effects of mild heat reduction, repair of allergic dermatitis damage, anti-inflammatory and detumescence and significant itching relief, has high bioavailability and good safety.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparation technology, and mainly relates to a prescription for a medical ointment and its preparation method, specifically to a prescription and preparation method for a medical antipyretic and antipruritic ointment. Background Technology

[0002] With the increasing demand for skin health protection among the public, problems such as fever due to external pathogens, itchy and hot skin, local redness, swelling and pain, and allergic dermatitis damage occur frequently. Although commonly used Western medicine preparations have a rapid onset of action, they have drawbacks such as strong irritation, easy recurrence, and unsuitability for long-term gentle external use. Traditional topical ointments generally have defects such as low bioavailability of active ingredients, poor compatibility and synergy, poor formability, and single function, making it difficult to meet the multiple needs of antipyretics, antipruritics, anti-swelling, and skin barrier repair at the same time.

[0003] Therefore, developing a medical antipyretic and antipruritic topical ointment based on the synergistic combination of modified polysaccharides, nano-drug-loaded complexes and immune adjuvants, with a reasonable formulation, optimized process, comprehensive effects, and mild and safe properties, has important practical application value. Summary of the Invention

[0004] In view of this, the present invention aims to overcome the shortcomings of the prior art and provide a prescription and preparation method for a medical antipyretic and antipruritic ointment, which has the effects of reducing fever, relieving itching, reducing swelling, and repairing skin.

[0005] In a first aspect, the present invention provides a medical antipyretic and antipruritic ointment, which is composed of functional active ingredients, base excipients and purified water, and the specific formula is as follows by mass percentage: Functional active ingredients: glycyrrhizic acid modified β-glucan 0.5%-2.0%, organoselenium-grafted sodium hyaluronate 1.0%-3.0%, hydroxypropyltrimethylammonium chloride chitosan-supported curcumin complex 1.0%-3.0%, polyinosinic acid (poly(I:C)) 0.05%-0.2%, CpG oligonucleotide 0.01%-0.05%, monophospholipid A (MPLA) 0.0005%-0.01%, peppermint oil 0.1%-0.5%; Matrix excipients: 0.5%-1.5% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 5.0%-12.0% glycerin, 2.0%-5.0% 1,2-pentanediol, 8.0%-15.0% white petrolatum, 2.0%-5.0% polyglycerol-3-methyl glucoside distearate, 2.0%-5.0% dimethicone, 5.0%-10.0% caprylic / capric triglyceride, 0.5%-1.0% phenoxyethanol; Purified water: Top up to 100%.

[0006] Furthermore, the formula for the medical antipyretic and antipruritic ointment is more specifically as follows: Functional active ingredients: glycyrrhizic acid modified β-glucan 1.5%-1.8%, organoselenium-grafted sodium hyaluronate 1.5%-2.0%, hydroxypropyltrimethylammonium chloride chitosan-supported curcumin complex 2.0%-2.5%, polyinosinic acid (poly(I:C)) 0.1%-0.15%, CpG oligonucleotide 0.032%-0.05%, monophospholipid A (MPLA) 0.008%-0.01%, peppermint oil 0.2%-0.4%; Matrix excipients: 0.8%-1.2% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 7.1%-9.4% glycerin, 3.0%-4.0% 1,2-pentanediol, 10.0%-12.0% white petrolatum, 4.0%-4.5% polyglycerol-3-methyl glucoside distearate, 3.0%-4.0% dimethicone, 6.06%-7.09% caprylic / capric triglyceride, 0.7%-0.9% phenoxyethanol; Purified water: Top up to 100%.

[0007] Preferably, the glycyrrhizic acid-modified β-glucan is a glycyrrhizic acid-modified β-glucan grafted with an acylhydrazine bond; Preparation method of glycyrrhizic acid-modified β-glucan grafted with hydrazide: Yeast β-glucan was dissolved in purified water to prepare a 6% (w / v) dispersion. After hydration by magnetic stirring and ultrasonic dispersion at 35℃, the mixture was cooled to 4℃ and the pH adjusted to 3.0-5.0. Sodium periodate (1%-2% of the dry weight of β-glucan) was added and reacted in the dark for 2-4 hours to oxidize and open the ring, introducing an aldehyde group. Ethylene glycol was added to quench the aldehyde group, and the aldehyde content was measured. Subsequently, adipate dihydrazide (ADH) was added in equimolar amounts to the aldehyde group, and the reaction was maintained at pH 5.0 for 4 hours to form a hydrazone bond. Sodium cyanoborohydride (1.5 times the molar amount of the aldehyde group) was then added, and the reaction was carried out at 4℃ for 2 hours to reduce the hydrazone bond to a stable secondary amine bond, retaining the free hydrazide group at the other end of the ADH. Glycyrrhizic acid (added at a molar ratio of 1.2:1 to the free hydrazide group of ADH) was then used in a 0.1% solution. Solubilize with mol / L sodium hydroxide solution, add EDC and NHS (molar ratio of 1:1, each amount being 1.2 times the molar amount of glycyrrhizic acid), adjust pH to 6.0, and activate at room temperature for 30 min to generate NHS ester; slowly add the activated solution dropwise to the above reaction solution containing free hydrazide groups, adjust pH to 6.0-6.5, and react at 37℃ for 12-24 h to complete hydrazide bond coupling (the consumption process of free hydrazide groups is monitored by ninhydrin colorimetric method); centrifuge or filter the reaction solution to remove insoluble matter, and put the filtrate into a dialysis bag with a molecular weight cutoff of 30000-50000 Da, and dialyze with purified water at 4℃ for 24-48 h (changing water 4-6 times). After freeze-drying, the dialysate is pulverized and passed through a 120-mesh sieve to obtain hydrazide-grafted glycyrrhizic acid modified β-glucan.

[0008] Preferably, the preparation method of the hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex in this invention is as follows: hydroxypropyltrimethylammonium chloride chitosan is dissolved in purified water to prepare a 2 mg / mL aqueous solution, and stirred until completely dissolved; 10 mg / mL curcumin ethanol solution is slowly added dropwise at a loading of 15%, and the mixture is sonicated at 250 W for 30 min to fully load and bind curcumin; during this process, the quaternary ammonium side chain of hydroxypropyltrimethylammonium chloride chitosan (HTCC) electrostatically enriches the polar groups (phenolic hydroxyl / β-diketenol) of curcumin, and the hydrophobic inclusion of the hydroxypropyl hydrophobic segment on the aromatic ring / diketone unit of curcumin, through cavitation shear confinement by 250 W sonication, self-assembles into nanoparticles with an average particle size of 185 nm; finally, it is purified by centrifugation at 3000 g for 10 min in a 100 kDa ultrafiltration centrifuge tube, the retentate is collected, freeze-dried, and then pulverized through an 80-mesh sieve to obtain the hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex.

[0009] Preferably, the preparation method of organoselenium-grafted sodium hyaluronate in this invention is as follows: Prepare a 7% aqueous solution of pharmaceutical-grade sodium hyaluronate, adjust the pH to 4.5-5.5, add 0.8 times the molar amount of EDC and an equimolar amount of NHS to the carboxyl group of hyaluronate, and activate at room temperature for 20 min; add 0.8% of the dry weight of sodium hyaluronate with selenomethionine, adjust the pH to 7.2, and react at room temperature for 8 h to form a stable amide bond; purify the product by dialysis using an 8000-12000 Da dialysis bag for 24 h, vacuum concentrate, spray dry, and pulverize through a 100-mesh sieve to obtain organoselenium-grafted sodium hyaluronate.

[0010] Preferably, the CpG oligonucleotide is selected from one of CpG oligonucleotide 1018, CpG oligonucleotide 1826, CpG oligonucleotide 2002, and CpG oligonucleotide 2395.

[0011] Secondly, the present invention provides a method for preparing a medical antipyretic and antipruritic ointment, comprising the following steps: S1: Aqueous phase preparation: Take 80% of the prescribed amount of purified water and add it to the emulsification reactor. Heat the mixture to 75°C. Add 80% of the prescribed amount of glycerol and the full amount of the prescribed amount of 1,2-pentanediol. Stir at 500-600 r / min for 10-15 min until completely dissolved. Keep the mixture at 75°C for later use.

[0012] S2: Oil phase preparation: Add the full prescription amount of white petrolatum, the full prescription amount of dimethicone, the full prescription amount of caprylic / capric triglyceride, and the full prescription amount of peppermint oil to the oil phase tank in sequence, heat to 80°C and stir until completely melted; add the full prescription amount of polyglycerol-3-methylglucose distearate, keep the temperature at 80°C and stir for 12-18 minutes until the system melts and becomes transparent to obtain a stable oil phase.

[0013] S3: Emulsification and homogenization: Under stirring conditions of 900-1100 r / min, slowly and uniformly add the oil phase to the water phase, maintain the system temperature at 75-80℃, and stir for 5 minutes to complete the initial mixing; turn on the high-pressure homogenizer and circulate the homogenizer 4-6 times under a pressure of 30MPa to refine it into a fine paste, and use cooling water to cool it down naturally.

[0014] S4: Low-Temperature Active Compound: When the temperature naturally drops below 40°C, add the full prescription amount of hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, and stir at 450-550 r / min for 15-20 min until fully hydrated and swollen. Then add the full prescription amount of organoselenium-grafted sodium hyaluronate and the full prescription amount of glycyrrhizic acid-modified β-glucan, and continue stirring for 10 min until evenly dispersed. Separately, take the full prescription amount of hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex and grind it with the remaining 20% ​​prescription amount of glycerin in a sterile mortar into a fine paste. When the paste cools to below 35°C, slowly add the paste into the system, adjust the stirring speed to 600 r / min, and continue stirring for 15-20 min until the mixture is smooth and free of impurities. The particles are visible to the naked eye. Simultaneously, the full prescription amount of poly(I:C) and the full prescription amount of CPG oligonucleotides are added to a pre-prepared MPLA pre-suspension (the MPLA pre-suspension is prepared by adding the full prescription amount of monophospholipid A to 20% of the prescription amount of purified water at 40°C, followed by ultrasonic dispersion). After stirring evenly under light-protected conditions, the mixture is slowly added to the paste in a thin stream, with the stirring speed controlled at 300-400 r / min for 15-20 min until the entire system is uniformly mixed. In this invention, the amount of nucleic acid-based immunoadjuvant is extremely low, forming a stable nanocomposite with the cationic carrier and amphiphilic polymers such as hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, resulting in no precipitation in the system.

[0015] S5: Finished product preparation: Add the full prescription amount of phenoxyethanol and mix at 400-500 r / min for 4-6 min; adjust the pH of the system to 5.1-5.9 using lactate-arginine buffer solution, and stir at low speed for 8-12 min under a negative pressure of -0.07--0.09 MPa to remove air bubbles inside the cream; place the cream in a constant temperature curing oven at 23-27℃ and let it stand for 24-36 h; perform aseptic filling in a clean area that meets the requirements of pharmaceutical GMP to obtain the finished product.

[0016] Preferably, the medical antipyretic and antipruritic ointment prepared by the above method has the effects of reducing fever, relieving itching, reducing swelling and repairing skin.

[0017] The present invention has the following beneficial effects: 1. The formula of this invention contains peppermint oil, whose cooling components can evaporate through the skin surface to remove heat, quickly relieve local fever and burning sensation, achieve gentle fever reduction and cooling, and soothe skin discomfort.

[0018] 2. The cream of this invention has excellent skin damage repair effects and can significantly improve symptoms of erythema, edema and epidermal peeling caused by allergic dermatitis. It can effectively repair damaged skin, but should be used with caution when the skin is broken over a large area.

[0019] 3. This invention has good anti-inflammatory and swelling-reducing effects, can effectively inhibit tissue swelling caused by acute inflammation, has a definite anti-inflammatory effect, and can quickly relieve discomfort such as skin redness and swelling.

[0020] 4. This invention has a significant antipruritic effect, can significantly inhibit acute itching reactions, reduce scratching behavior, and effectively relieve skin allergies, itching and discomfort.

[0021] 5. The formulation of this invention is scientifically innovative, employing modified polysaccharides, nano-drug-loaded complexes, multiple functional active components, and triple immune adjuvants in synergistic combination, while also incorporating peppermint oil. The components work together to enhance the overall efficacy. The preparation process uses high-pressure homogenization combined with low-temperature segmented active compounding technology, which ensures that the ointment has a uniform and delicate texture and good physicochemical stability, while maximizing the preservation of the bioactivity of the modified components, drug-loaded complexes, and immune adjuvants. The product quality is stable and reliable, and its comprehensive application value is high.

[0022] 6. Mechanism of action: The medical antipyretic and antipruritic ointment of the present invention is not a simple superposition of the components, but a synergistic design based on physical antipyresis, barrier repair and immune regulation.

[0023] Firstly, the physical evaporation and heat dissipation of peppermint oil, combined with the deep anti-inflammatory and swelling-reducing effects of the hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex, achieves a holistic treatment, addressing both the symptoms and the underlying cause. Secondly, glycyrrhizic acid-modified β-glucan and organic selenium-grafted sodium hyaluronate, as modified polysaccharide carriers, not only have their own moisturizing and repairing functions, but also promote the presentation of immune adjuvants on the skin, activate local immune defense, and relieve the symptoms of allergic dermatitis from the root cause. Third, the specific ratio of the triple immune adjuvant to the modified polysaccharide produced an unexpected synergistic effect, significantly reducing the irritation that a single adjuvant might cause, while enhancing the overall antipruritic and anti-swelling efficacy. Attached Figure Description

[0024] To more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0025] Figure 1 A comparison chart of skin damage scores for mice in each group.

[0026] Figure 2 A comparison of ear swelling in each group of mice.

[0027] Figure 3 A comparison chart showing the number of scratches by mice in each group.

[0028] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that, unless otherwise specified, the following embodiments and features described therein can be combined with each other.

[0029] Example 1

[0030] This embodiment provides a medical antipyretic and antipruritic ointment, the specific steps of which are as follows: The specific prescription for the medical antipyretic and antipruritic ointment is as follows: Functional active ingredients: glycyrrhizic acid modified β-glucan 1.5%, organoselenium-grafted sodium hyaluronate 1.5%, hydroxypropyltrimethylammonium chloride chitosan-supported curcumin complex 2.0%, polyinosinic acid (poly(I:C)) 0.1%, CpG oligonucleotide 1018 0.032%, monophospholipid A (MPLA) 0.008%, peppermint oil 0.2%; Matrix excipients: 0.8% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 7.1% glycerin, 3.0% 1,2-pentanediol, 10.0% white petrolatum, 4.0% polyglycerol-3-methyl glucoside distearate, 3.0% dimethicone, 6.06% caprylic / capric triglyceride, 0.7% phenoxyethanol; Purified water: Top up to 100%.

[0031] S1: Aqueous phase preparation: Take 80% of the prescribed amount of purified water and add it to the emulsification reactor. Heat the mixture to 75°C. Add 80% of the prescribed amount of glycerol and the full amount of the prescribed amount of 1,2-pentanediol. Stir at 500 r / min for 10 min until completely dissolved. Keep the mixture at 75°C for later use.

[0032] S2: Oil phase preparation: Add the full prescription amount of white petrolatum, the full prescription amount of dimethicone, the full prescription amount of caprylic / capric triglyceride, and the full prescription amount of peppermint oil to the oil phase tank in sequence, heat to 80°C and stir until completely melted; add the full prescription amount of polyglycerol-3-methylglucose distearate, keep the temperature at 80°C and stir for 12 minutes until the system melts and becomes transparent to obtain a stable oil phase.

[0033] S3: Emulsification and homogenization: Under stirring conditions of 900 r / min, the oil phase is slowly and uniformly added to the water phase, the system temperature is maintained at 75℃, and the mixture is stirred for 5 min to complete the initial mixing; the high-pressure homogenizer is turned on and the homogenizer is circulated and homogenized 4 times under a pressure of 30 MPa to refine the paste into a fine paste, and then cooled naturally with cooling water.

[0034] S4: Low-temperature active compound: When the temperature naturally drops below 40°C, add the full prescription amount of hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, and stir at 450 r / min for 15 min until fully hydrated and swollen. Then add the full prescription amount of organoselenium-grafted sodium hyaluronate and the full prescription amount of glycyrrhizic acid-modified β-glucan, and continue stirring for 10 min until evenly dispersed. Separately, take the full prescription amount of hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex and grind it with the remaining 20% ​​prescription amount of glycerin in a sterile mortar into a fine paste. When the paste cools to below 35°C, slowly add the paste to the mixture. Add the contents to the system, adjust the stirring speed to 600 r / min, and continue stirring for 15 min until no visible particles are present. At the same time, add the full amount of poly(I:C) and the full amount of CpG oligonucleotide 1018 to the pre-prepared MPLA pre-suspension (the MPLA pre-suspension is prepared by adding the full amount of monophospholipid A to 20% of the prescribed amount of purified water at 40℃ and then dispersing it thoroughly by ultrasound). Stir evenly under light-protected conditions and then slowly add the contents to the paste in a thin stream. Adjust the stirring speed to 300 r / min and stir at low speed for 15 min until the entire system is mixed evenly.

[0035] S5: Finished product preparation: Add the full prescription amount of phenoxyethanol and mix at 400 r / min for 4 min; adjust the pH of the system to 5.1 using lactate-arginine buffer solution, and stir at low speed for 8 min under a negative pressure of -0.07 MPa to remove air bubbles inside the cream; place the cream in a 23℃ constant temperature curing oven and let it stand for 24 h; perform aseptic filling in a clean area that meets the requirements of pharmaceutical GMP to obtain the finished product.

[0036] The specific preparation method of glycyrrhizic acid-modified β-glucan grafted with hydrazide is as follows: Yeast β-glucan is mixed with purified water to prepare a 6% (w / v) dispersion. After hydration by magnetic stirring and ultrasonication at 35℃, the mixture is cooled to 4℃ and the pH is adjusted to 3.0. Sodium periodate (1% of the dry weight of β-glucan) is added and reacted in the dark for 2 hours to oxidize and open the ring, introducing an aldehyde group. Ethylene glycol is added to quench the aldehyde group, and the aldehyde content is measured. Adipic acid dihydrazide (ADH) with an equimolar amount of aldehyde group is added, and the reaction is maintained at pH 5.0 for 4 hours to form a hydrazone bond. Sodium cyanoborohydride (1.5 times the molar amount of aldehyde group) is then added and reacted at 4℃ for 2 hours to reduce the hydrazone bond to a stable secondary amine bond, retaining the free hydrazide group at the other end of the ADH. Glycyrrhizic acid (added at a molar ratio of 1.2:1 to the free hydrazide group of ADH) is then used in a 0.1% solution. Solubilize with mol / L sodium hydroxide solution, add EDC and NHS (molar ratio of 1:1, each amount being 1.2 times the molar amount of glycyrrhizic acid), adjust pH to 6.0, and activate at room temperature for 30 min to generate NHS ester; slowly add the activated solution dropwise to the above reaction solution containing free hydrazide groups, adjust pH to 6.0, and react at 37℃ for 12 h to complete hydrazide bond coupling (the consumption process of free hydrazide groups is monitored by ninhydrin colorimetric method); centrifuge or filter the reaction solution to remove insoluble matter, and put the filtrate into a dialysis bag with a molecular weight cutoff of 30000 Da, and dialyze with purified water at 4℃ for 24 h (water changed 4 times). After freeze-drying, the dialysate is pulverized and passed through a 120-mesh sieve to obtain hydrazide-grafted glycyrrhizic acid modified β-glucan.

[0037] The preparation method of hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex is as follows: hydroxypropyltrimethylammonium chloride chitosan (purchased from Shanghai Yuanye Biotechnology Co., Ltd.) is added to purified water to prepare a 2 mg / mL aqueous solution, and stirred until completely dissolved; 10 mg / mL curcumin ethanol solution is slowly added dropwise at a loading of 15%, and the mixture is sonicated at 250W for 30 min to allow curcumin to be fully loaded and bound; the mixture is purified by centrifugation at 3000g for 10 min using a 100 kDa ultrafiltration centrifuge tube, and the retentate is collected, freeze-dried, and then pulverized through an 80-mesh sieve to obtain the hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex.

[0038] The specific preparation method of organoselenium-grafted sodium hyaluronate is as follows: Prepare a 7% aqueous solution of pharmaceutical-grade sodium hyaluronate, adjust the pH to 4.5, add 0.8 times the molar amount of EDC and an equimolar amount of NHS to the carboxyl group of hyaluronate, and activate at room temperature for 20 min; add 0.8% of the dry weight of sodium hyaluronate with selenomethionine, adjust the pH to 7.2, and react at room temperature for 8 h to form a stable amide bond; purify the product by dialysis with an 8000 Da dialysis bag for 24 h, vacuum concentrate, spray dry, and pulverize through a 100-mesh sieve to obtain organoselenium-grafted sodium hyaluronate.

[0039] Example 2

[0040] This embodiment provides a medical antipyretic and antipruritic ointment, the specific steps of which are as follows: The specific prescription for the medical antipyretic and antipruritic ointment is as follows: Functional active ingredients: glycyrrhizic acid modified β-glucan 1.7%, organoselenium-grafted sodium hyaluronate 1.8%, hydroxypropyltrimethylammonium chloride chitosan-supported curcumin complex 2.3%, polyinosinic acid (poly(I:C)) 0.13%, CpG oligonucleotide 1018 0.04%, monophospholipid A (MPLA) 0.009%, peppermint oil 0.3%; Matrix excipients: 1.0% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 8.2% glycerin, 3.5% 1,2-pentanediol, 11.0% white petrolatum, 4.3% polyglycerol-3-methyl glucoside distearate, 3.5% dimethicone, 6.6% caprylic / capric triglyceride, 0.8% phenoxyethanol; Purified water: Top up to 100%.

[0041] S1: Aqueous phase preparation: Take 80% of the prescribed amount of purified water and add it to the emulsification reactor. Heat the mixture to 75°C. Add 80% of the prescribed amount of glycerol and the full amount of the prescribed amount of 1,2-pentanediol. Stir at 550 r / min for 13 min until completely dissolved. Keep the mixture at 75°C for later use.

[0042] S2: Oil phase preparation: Add the full prescription amount of white petrolatum, the full prescription amount of dimethicone, the full prescription amount of caprylic / capric triglyceride, and the full prescription amount of peppermint oil to the oil phase tank in sequence, heat to 80°C and stir until completely melted; add the full prescription amount of polyglycerol-3-methylglucose distearate, keep the temperature at 80°C and stir for 15 minutes until the system melts and becomes transparent to obtain a stable oil phase.

[0043] S3: Emulsification and homogenization: Under stirring conditions of 1000 r / min, the oil phase is slowly and uniformly added to the water phase, the system temperature is maintained at 78℃, and the mixture is stirred for 5 minutes to complete the initial mixing; the high-pressure homogenizer is turned on and the homogenizer is circulated and homogenized 5 times under a pressure of 30 MPa to refine the paste into a fine paste, and then cooled naturally with cooling water.

[0044] S4: Low-temperature active compound: When the temperature naturally drops below 40°C, add the full prescription amount of hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, and stir at 500 r / min for 18 min until fully hydrated and swollen. Then add the full prescription amount of organoselenium-grafted sodium hyaluronate and the full prescription amount of glycyrrhizic acid-modified β-glucan, and continue stirring for 10 min until evenly dispersed. Separately, take the full prescription amount of hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex and grind it with the remaining 20% ​​prescription amount of glycerin in a sterile mortar into a fine paste. When the paste cools to below 35°C, slowly add the paste to the mixture. Add the mixture to the system, adjust the stirring speed to 600 r / min, and continue stirring for 18 min until no visible particles are present. At the same time, add the full amount of poly(I:C) and the full amount of CpG oligonucleotide 1018 to the pre-prepared MPLA pre-suspension (the MPLA pre-suspension is prepared by adding the full amount of monophospholipid A to 20% of the prescribed amount of purified water at 40℃ and then dispersing it thoroughly by ultrasound). Stir evenly under light-protected conditions and then slowly add the mixture to the paste in a thin stream. Adjust the stirring speed to 350 r / min and stir at low speed for 18 min until the entire system is mixed evenly.

[0045] S5: Finished product preparation: Add the full prescription amount of phenoxyethanol and mix at 450 r / min for 5 min; adjust the pH of the system to 5.5 using lactate-arginine buffer solution, and stir at low speed for 10 min under a negative pressure of -0.08 MPa to remove air bubbles inside the cream; place the cream in a 25℃ constant temperature curing oven and let it stand for 30 h; perform aseptic filling in a clean area that meets the requirements of pharmaceutical GMP to obtain the finished product.

[0046] The specific preparation method of glycyrrhizic acid-modified β-glucan grafted with hydrazide is as follows: Yeast β-glucan is mixed with purified water to prepare a 6% (w / v) dispersion. After magnetic stirring and ultrasonic dispersion at 35℃, the mixture is cooled to 4℃ and the pH is adjusted to 3.0-5.0. Sodium periodate (1.5% of the dry weight of β-glucan) is added and reacted in the dark for 3 hours to oxidize and open the ring, introducing an aldehyde group. Ethylene glycol is added to quench the aldehyde group, and the aldehyde content is measured. Adipic acid dihydrazide (ADH) with an equimolar amount of aldehyde group is added, and the reaction is maintained at pH 5.0 for 4 hours to form a hydrazone bond. Sodium cyanoborohydride (1.5 times the molar amount of aldehyde group) is then added and reacted at 4℃ for 2 hours to reduce the hydrazone bond to a stable secondary amine bond, retaining the free hydrazide group at the other end of the ADH. Glycyrrhizic acid (added at a molar ratio of 1.2:1 to the free hydrazide group of ADH) is then used in a 0.1% solution. Solubilize with mol / L sodium hydroxide solution, add EDC and NHS (molar ratio of 1:1, each amount being 1.2 times the molar amount of glycyrrhizic acid), adjust pH to 6.0, and activate at room temperature for 30 min to generate NHS ester; slowly add the activated solution dropwise to the above reaction solution containing free hydrazide groups, adjust pH to 6.3, and react at 37℃ for 18 h to complete hydrazide bond coupling (the consumption process of free hydrazide groups is monitored by ninhydrin colorimetric method); centrifuge or filter the reaction solution to remove insoluble matter, and put the filtrate into a dialysis bag with a molecular weight cutoff of 40000 Da, and dialyze with purified water at 4℃ for 30 h (water changed 5 times). After freeze-drying, the dialysate is pulverized and passed through a 120-mesh sieve to obtain hydrazide-grafted glycyrrhizic acid modified β-glucan.

[0047] The preparation method of hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex is as follows: hydroxypropyltrimethylammonium chloride chitosan (purchased from Shanghai Yuanye Biotechnology Co., Ltd.) is added to purified water to prepare a 2 mg / mL aqueous solution, and stirred until completely dissolved; 10 mg / mL curcumin ethanol solution is slowly added dropwise at a loading of 15%, and the mixture is sonicated at 250W for 30 min to allow curcumin to be fully loaded and bound; the mixture is purified by centrifugation at 3000g for 10 min using a 100 kDa ultrafiltration centrifuge tube, and the retentate is collected, freeze-dried, and then pulverized through an 80-mesh sieve to obtain the hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex.

[0048] The specific preparation method of organoselenium-grafted sodium hyaluronate is as follows: Prepare a 7% aqueous solution of pharmaceutical-grade sodium hyaluronate, adjust the pH to 5, add 0.8 times the molar amount of EDC and an equimolar amount of NHS to the carboxyl group of hyaluronate, and activate at room temperature for 20 min; add 0.8% of the dry weight of sodium hyaluronate with selenomethionine, adjust the pH to 7.2, and react at room temperature for 8 h to form a stable amide bond; purify the product by dialysis with a 10000 Da dialysis bag for 24 h, concentrate under vacuum, spray dry, and pulverize through a 100-mesh sieve to obtain organoselenium-grafted sodium hyaluronate.

[0049] Example 3

[0050] This embodiment provides a medical antipyretic and antipruritic ointment, the specific steps of which are as follows: The specific prescription for the medical antipyretic and antipruritic ointment is as follows: Functional active ingredients: glycyrrhizic acid modified β-glucan 1.8%, organoselenium-grafted sodium hyaluronate 2.0%, hydroxypropyltrimethylammonium chloride chitosan-supported curcumin complex 2.5%, polyinosinic acid (poly(I:C)) 0.15%, CpG oligonucleotide 1018 0.05%, monophospholipid A (MPLA) 0.01%, peppermint oil 0.4%; Matrix excipients: 1.2% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 9.4% glycerin, 4.0% 1,2-pentanediol, 12.0% white petrolatum, 4.5% polyglycerol-3-methyl glucoside distearate, 4.0% dimethicone, 7.09% caprylic / capric triglyceride, and 0.9% phenoxyethanol; Purified water: Top up to 100%.

[0051] S1: Aqueous phase preparation: Take 80% of the prescribed amount of purified water and add it to the emulsification reactor. Heat the mixture to 75°C. Add 80% of the prescribed amount of glycerol and the full amount of the prescribed amount of 1,2-pentanediol. Stir at 600 r / min for 15 min until completely dissolved. Keep the mixture at 75°C for later use.

[0052] S2: Oil phase preparation: Add the full prescription amount of white petrolatum, the full prescription amount of dimethicone, the full prescription amount of caprylic / capric triglyceride, and the full prescription amount of peppermint oil to the oil phase tank in sequence, heat to 80°C and stir until completely melted; add the full prescription amount of polyglycerol-3-methylglucose distearate, keep the temperature at 80°C and stir for 18 minutes until the system melts and becomes transparent to obtain a stable oil phase.

[0053] S3: Emulsification and homogenization: Under stirring conditions of 1100 r / min, the oil phase is slowly and uniformly added to the water phase, the system temperature is maintained at 80℃, and the initial mixing is completed by stirring for 5 minutes; the high-pressure homogenizer is turned on, and the homogenizer is circulated and homogenized 6 times under a pressure of 30 MPa to refine it into a fine paste, and then cooled naturally with cooling water.

[0054] S4: Low-Temperature Active Compound: When the temperature naturally drops below 40°C, add the full prescription amount of hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, and stir at 550 r / min for 20 min until fully hydrated and swollen. Then add the full prescription amount of organoselenium-grafted sodium hyaluronate and the full prescription amount of glycyrrhizic acid-modified β-glucan, and continue stirring for 10 min until evenly dispersed. Separately, take the full prescription amount of hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex and grind it with the remaining 20% ​​prescription amount of glycerin in a sterile mortar into a fine paste. When the paste cools to below 35°C, slowly add the paste to the mixture. Add the contents to the system, adjust the stirring speed to 600 r / min, and continue stirring for 20 min until no visible particles are present. At the same time, add the full amount of poly(I:C) and the full amount of CpG oligonucleotide 1018 to the pre-prepared MPLA pre-suspension (the MPLA pre-suspension is prepared by adding the full amount of monophospholipid A to 20% of the prescribed amount of purified water at 40℃ and then dispersing it thoroughly by ultrasound). Stir evenly under light-protected conditions and then slowly add the contents to the paste in a thin stream. Adjust the stirring speed to 400 r / min and stir at low speed for 20 min until the entire system is mixed evenly.

[0055] S5: Finished product preparation: Add the full prescription amount of phenoxyethanol and mix at 500 r / min for 6 min; adjust the pH of the system to 5.9 using lactate-arginine buffer solution, and stir at low speed for 12 min under a negative pressure of -0.09 MPa to remove air bubbles inside the cream; place the cream in a 27℃ constant temperature curing oven and let it stand for 36 h; perform aseptic filling in a clean area that meets the requirements of pharmaceutical GMP to obtain the finished product.

[0056] In this process, yeast β-glucan was prepared into a 6% (w / v) dispersion with purified water. After hydration by magnetic stirring and ultrasonication at 35℃, the mixture was cooled to 4℃ and the pH adjusted to 5.0. Sodium periodate (2% of the dry weight of β-glucan) was added and reacted in the dark for 4 hours to oxidize and open the ring, introducing an aldehyde group. Ethylene glycol was added to quench the aldehyde group, and the aldehyde content was measured. Adipic acid dihydrazide (ADH) of equal molar amount to the aldehyde group was added, and the reaction was maintained at pH 5.0 for 4 hours to form a hydrazone bond. Then, sodium cyanoborohydride (1.5 times the molar amount of the aldehyde group) was added and reacted at 4℃ for 2 hours to reduce the hydrazone bond to a stable secondary amine bond, retaining the free hydrazide group at the other end of the ADH. Glycyrrhizic acid (added at a molar ratio of 1.2:1 to the free hydrazide group of ADH) was used in a 0.1% reaction. Solubilize with mol / L sodium hydroxide solution, add EDC and NHS (molar ratio of 1:1, each amount being 1.2 times the molar amount of glycyrrhizic acid), adjust pH to 6.0, and activate at room temperature for 30 min to generate NHS ester; slowly add the activated solution dropwise to the above reaction solution containing free hydrazide groups, adjust pH to 6.5, and react at 37℃ for 24 h to complete hydrazide bond coupling (the consumption process of free hydrazide groups is monitored by ninhydrin colorimetric method); centrifuge or filter the reaction solution to remove insoluble matter, and put the filtrate into a dialysis bag with a molecular weight cutoff of 50000 Da, and dialyze with purified water at 4℃ for 48 h (water changed 6 times). After freeze-drying, the dialysate is pulverized and passed through a 120-mesh sieve to obtain hydrazide-grafted glycyrrhizic acid modified β-glucan.

[0057] The preparation method of hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex is as follows: hydroxypropyltrimethylammonium chloride chitosan (purchased from Shanghai Yuanye Biotechnology Co., Ltd.) is added to purified water to prepare a 2 mg / mL aqueous solution, and stirred until completely dissolved; 10 mg / mL curcumin ethanol solution is slowly added dropwise at a loading of 15%, and the mixture is sonicated at 250W for 30 min to allow curcumin to be fully loaded and bound; the mixture is purified by centrifugation at 3000g for 10 min using a 100 kDa ultrafiltration centrifuge tube, and the retentate is collected, freeze-dried, and then pulverized through an 80-mesh sieve to obtain the hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex.

[0058] The specific preparation method of organoselenium-grafted sodium hyaluronate is as follows: Prepare a 7% aqueous solution of pharmaceutical-grade sodium hyaluronate, adjust the pH to 5.5, add 0.8 times the molar amount of EDC and an equimolar amount of NHS to the carboxyl group of hyaluronate, and activate at room temperature for 20 min; add 0.8% of the dry weight of sodium hyaluronate with selenomethionine, adjust the pH to 7.2, and react at room temperature for 8 h to form a stable amide bond; purify the product by dialysis with a 12000 Da dialysis bag for 24 h, concentrate under vacuum, spray dry, and pulverize through a 100-mesh sieve to obtain organoselenium-grafted sodium hyaluronate.

[0059] Comparative Example 1: The preparation process is the same as in Example 2, except that polyinosinic acid is not added in step S4. The remaining raw material ratios, process parameters and operating steps are completely consistent with those in Example 2.

[0060] Comparative Example 2: The preparation process is the same as in Example 2, except that CpG oligonucleotide 1018 is not added in step S4. The other raw material ratios, process parameters and operating steps are completely consistent with those in Example 2.

[0061] Comparative Example 3: The preparation process is the same as in Example 2, except that monophospholipid A is not added in step S4. The remaining raw material ratios, process parameters and operating steps are completely consistent with those in Example 2.

[0062] Comparative Example 4: The preparation process is the same as in Example 2, except that glycyrrhizic acid-modified β-glucan grafted with hydrazide bonds is not added in step S4. The remaining raw material ratios, process parameters and operating steps are completely consistent with those in Example 2.

[0063] Comparative Example 5: The preparation process is the same as in Example 2, except that in step S4, the glycyrrhizic acid-modified β-glucan grafted with hydrazide bonds is replaced with an equal mass of unmodified yeast β-glucan. The remaining raw material ratios, process parameters and operating steps are completely consistent with those in Example 2.

[0064] Comparative Example 6: The preparation process is the same as in Example 2, except that hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex is not added in step S4. The remaining raw material ratios, process parameters and operating steps are completely consistent with those in Example 2.

[0065] Comparative Example 7: The preparation process was the same as in Example 2, except that in step S4, equal masses of free curcumin and hydroxypropyltrimethylammonium chloride chitosan were used instead of hydroxypropyltrimethylammonium chloride chitosan to support the curcumin complex. Specifically, 0.3g of curcumin (equivalent to the 15% drug loading of the 2.0g complex in Example 2) and 1.7g of hydroxypropyltrimethylammonium chloride chitosan were weighed out. 0.3g of curcumin was completely dissolved in a small amount of anhydrous ethanol and then mixed with 1.7g of blank chitosan and 1.6g of glycerol. The mixture was then ground into a uniform paste in a sterile mortar. When the paste cooled to below 35°C, the physically mixed paste was slowly added to the ointment. The remaining raw material ratios, process parameters, and operating steps were completely consistent with Example 2.

[0066] Comparative Example 8: The preparation process is the same as in Example 2, except that in step S4, only the organic selenium-grafted sodium hyaluronate is not added. The other raw material ratios, process parameters and operating steps are completely consistent with those in Example 2.

[0067] Comparative Example 9: The preparation process is the same as in Example 2, except that in step 4, an equal mass of unmodified sodium hyaluronate is used instead of organic selenium-grafted sodium hyaluronate. The remaining raw material ratios, process parameters and operating steps are completely consistent with those in Example 2.

[0068] Experiment 1: Characterization Test The three modified materials prepared in Example 2 were characterized structurally and their physicochemical properties were determined. The specific data and conclusions are as follows. 1. Glycyrrhizic acid (GA) modified β-glucan grafted with hydrazide bonds The structure was confirmed using ¹H NMR and Fourier transform infrared spectroscopy (FT-IR).

[0069] ¹H NMR analysis ( (9:1, v / v): Unmodified β-glucan only showed proton signals in the sugar ring at δ 3.0~4.0 ppm; the modified product spectrum showed characteristic angular methyl proton peaks of glycyrrhizic acid triterpenoid nuclei in the δ 0.82~1.15 ppm range, and characteristic olefinic hydrogen peaks on the C=C double bond of glycyrrhizic acid appeared near δ 5.5~6.0 ppm, directly proving that the glycyrrhizic acid structural unit has been successfully introduced into the polysaccharide backbone.

[0070] FT-IR analysis showed that the modified product was within the range of 1660–1680 cm⁻¹. - The characteristic absorption peak of the hydrazide bond C=O (amide I band) appears at 1545 cm⁻¹. - ¹ Near the amide II band (NH bending vibration) absorption peak; simultaneously, the C=O absorption of the residual aldehyde group in oxidized dextran (~1720 cm⁻¹) is observed. - ¹) The absorption of the C=N hydrazone bond was almost completely eliminated, and no obvious unreduced characteristic absorption was detected, indicating that... The reduction reaction has converted the intermediate hydrazone bond into a stable secondary amine bond, and the hydrazide coupling reaction between glycyrrhizic acid and the polysaccharide backbone has been successfully completed.

[0071] Grafting rate was determined by UV-Vis spectrophotometry. A standard curve (R²>0.999) was plotted at 251 nm using glycyrrhizic acid standard. The modified product was dissolved in PBS (pH 7.4) with ultrasonic assistance and then measured at the same wavelength. The calculated content of covalently bound glycyrrhizic acid was 9.02%. Based on the preparation feed chain, the grafting rate was calculated as follows: Based on the feed amount (1.5% of dextran dry weight), the theoretical upper limit of GA grafting is estimated to be 13.84%. The measured value reached 65.2% of this theoretical value. The limitation is mainly attributed to the dense triple helix crystalline region of yeast β-glucan. The steric hindrance effect of infiltration and subsequent grafting of high molecular weight glycyrrhizic acid was investigated; however, the results confirm the robust feasibility of the two-step process of oxidative ring-opening and hydrazide coupling at available apparent sites.

[0072] 2. Hydroxypropyltrimethylammonium chloride chitosan (HTCC) loaded with curcumin (Cur) complex Physical state and structure were confirmed using X-ray diffraction (XRD) and dynamic light scattering (DLS).

[0073] XRD analysis showed that pure curcumin exhibited multiple sharp crystalline diffraction peaks at 2θ of 8.7°, 17.4°, and 25.5°, while the pure HTCC support showed typical amorphous diffuse peaks. After loading curcumin, the characteristic crystalline diffraction peaks of curcumin in the complex spectrum completely disappeared, showing only broadened amorphous diffuse peaks, indicating that curcumin no longer exists in a crystalline form in the support system, but is effectively encapsulated in an amorphous or highly molecularly dispersed state.

[0074] DLS and Zeta potential measurements showed that the average particle size of the complex was 185.4 nm, the polydispersity index (PDI) was 0.216, and the Zeta potential was +38.6 mV. The high positive potential confirmed the retention of the HTCC quaternary ammonium salt framework; the complex was stably dispersed through a combination of electrostatic repulsion between like-charged colloids (anti-agglomeration between particles) and electrostatic enrichment / hydrophobic inclusion between HTCC-Cur particles.

[0075] Drug loading and encapsulation efficiency were determined by UV-Vis spectrophotometry at 425 nm. The results showed that the actual drug loading of the complex was 12.85%, and the encapsulation efficiency was 85.67%, meeting the high drug loading requirements of the formulation design. The high drug loading and encapsulation efficiency were mainly attributed to the electrostatic enrichment of curcumin molecules by the HTCC quaternary ammonium cation field, and the synergistic hydrophobic inclusion of curcumin aromatic rings / diketone units by the hydroxypropyl hydrophobic segment. The preparation process was stable and controllable.

[0076] 3. Organic selenium-grafted sodium hyaluronate The selenium content was accurately determined using inductively coupled plasma mass spectrometry (ICP-MS). The sample was digested by microwave and then... The sample was tested using the detection channel. The results showed that the organic selenium content in the product was 2.48 mg / g, which is approximately 77.0% of the theoretical upper limit of selenium (3.22 mg / g) calculated based on the feed amount (0.8% of selenomethionine by dry weight of sodium hyaluronate). This indicates that selenomethionine has been introduced into the side chain of sodium hyaluronate, and the grafting reaction efficiency is good.

[0077] The structure was confirmed using X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FT-IR).

[0078] XPS full-spectrum scanning confirmed the presence of C, N, O, and Se elements in the product; the high-resolution Se 3d spectrum showed a distinct characteristic peak at a binding energy of 55.2 eV, corresponding to the C-Se-C structure in the organoselenide, directly proving that the selenium element was successfully incorporated into the HA framework in the form of covalent bonds, ruling out the possibility of physical adsorption or inorganic selenium residue.

[0079] FT-IR analysis showed that, compared with pure HA, the modified product showed a lower concentration at 1650 cm⁻¹. - ¹(Amide I band) and 1550 cm - The absorption peak intensity at ¹ (amide II band) is significantly enhanced, and at 1320 cm⁻¹ - ¹A weak Se-C stretching vibration shoulder was observed nearby. Combined with XPS results, this confirmed that the α-amino group of selenomethionine reacted with the carboxyl group of sodium hyaluronate to form a stable covalent amide bond.

[0080] Experiment 2: Animal Experiments on the Effect of Repairing the Skin Barrier Animals: SPF-grade male BALB / c mice, weighing 18-22g, were purchased from Beijing Speford Biotechnology Co., Ltd. Before the experiment officially began, all mice underwent a 7-day environmental acclimatization period. During this period, the temperature of the housing was maintained at 25±2℃, the humidity was controlled at 55%-65%, and a 12-hour light-dark cycle was provided. Mice were allowed free access to water during this time.

[0081] The day before the experiment, the mice were shaved on their backs using an electric shaver. The area to be shaved was 2cm × 3cm.

[0082] Grouping: Mice were randomly divided into normal group, model group, positive control group, blank matrix group (functional active ingredients were removed, matrix excipients were retained, the preparation process was the same as in Example 2, and the following experiments were the same), Example 1-3 groups, comparative examples 1-5 groups and comparative examples 8-9 groups, with 10 mice in each group.

[0083] Modeling: Except for the normal group, all other groups of mice were modeled. Specifically, on days 1-2 of the experiment, 25 μL of 7% 2,4-dinitrochlorobenzene solution (acetone-olive oil, volume ratio 4:1) was evenly applied to the hairless area on the back of the mice to complete the sensitization. On days 5, 8, and 11 of the experiment, 20 μL of 1% dinitrochlorobenzene solution was applied to the same area for challenge. Mice in the normal group were simultaneously applied with an equal amount of acetone-olive oil mixture without dinitrochlorobenzene.

[0084] Administration: Administer medication starting from day 1 of the experiment.

[0085] Normal group: No modeling or drug administration.

[0086] Model group: No medication is administered after modeling.

[0087] Positive control group: Apply 0.5g of mometasone furoate cream per animal once a day for 14 consecutive days.

[0088] Blank matrix group: Apply 0.2g of blank matrix per animal once a day for 14 consecutive days.

[0089] Examples 1-3, Comparative Examples 1-5 and Comparative Examples 8-9: The corresponding triple-targeted modified antipruritic and anti-swelling cream was evenly applied to the hair removal area daily, with a dosage of 0.2g / animal, once a day, for 14 consecutive days.

[0090] 4.1 Skin damage severity score Twenty-four hours after the last administration, the erythema, edema, epidermal shedding, and scratches on the backs of mice in each group were observed. The skin damage scoring criteria (Table 1) were used for quantitative scoring. The experimental results are shown in Table 2 and... Figure 1 As shown.

[0091] Table 1

[0092] Table 2

[0093] Table 2 shows the comparison of skin damage scores in the 2,4-dinitrochlorobenzene-induced skin injury model in each group of mice. The results showed that the skin damage severity in the model groups was significantly higher than that in the normal group. Compared with the model group, the skin damage severity in Examples 1-3 and each comparative group was reduced to varying degrees, with Example 2 showing the lowest skin damage severity. The skin damage severity in Examples 1-3 was significantly lower than that in each comparative group, indicating that the medical antipyretic and antipruritic ointment of this invention has significant skin damage repair and anti-inflammatory effects. Furthermore, the synergistic effect of glycyrrhizic acid-modified β-glucan, organoselenium-grafted sodium hyaluronate, and the immune adjuvant is crucial to the repair effect. Removing key modified components or omitting the modification treatment will significantly weaken the skin repair efficacy; however, caution should be exercised when using this ointment on large areas of broken skin.

[0094] Comparative data show that removing any of the immune adjuvants (comparative examples 1-3) leads to a sharp decrease in efficacy. The mechanism lies in the complementary regulatory effects of poly(I:C), CpG, and MPLA, which act as TLR3, TLR9, and TLR4 agonists, respectively, by synergistically reshaping the local immune microenvironment of the skin lesion: MPLA drives the maturation of dendritic cells and macrophages and promotes Th1 immune shift, thereby antagonizing the Th2 inflammatory dominance induced by DNCB; CpG activates B cells and NK cells, inducing IFN-γ secretion to further inhibit Th2 inflammatory factors such as IL-4; and poly(I:C) enhances the type I interferon response, promoting the cascade amplification of local anti-inflammatory and tissue repair signals. Through synergistic intervention via different pathways, these three agents shift severe allergic inflammation to a controllable repair state, a comprehensive immune correction that cannot be achieved by a single component.

[0095] Meanwhile, compared to the use of unmodified β-glucan (Comparative Examples 4 and 5), glycyrrhizic acid-modified β-glucan, covalently grafted with hydrazide bonds, utilizes the specific recognition and endocytosis pathway between β-glucan and the Dectin-1 receptor on the surface of macrophages to target and deliver covalently bound glycyrrhizic acid into the interior of inflammation-related immune cells, thereby overcoming the defects of low transdermal efficiency and insufficient local accumulation of free glycyrrhizic acid. Grafted glycyrrhizic acid inhibits the pro-inflammatory cascade by directly blocking the HMGB1 / TLR4 signaling axis, and at the same time, it locally increases endogenous cortisol levels by inhibiting 11β-hydroxysteroid dehydrogenase activity, thus synergistically enhancing the local anti-inflammatory effect.

[0096] Furthermore, compared to sodium hyaluronate that is absent or unmodified (Comparative Examples 8 and 9), organoselenium-grafted sodium hyaluronate enhances the local retention and accumulation of selenomethionine in the inflammatory wound area by leveraging the specific affinity of hyaluronic acid for the highly expressed CD44 receptor at the lesion site. This allows it to act as a core cofactor of glutathione peroxidase, continuously clearing excessive reactive oxygen species and blocking secondary tissue damage caused by oxidative stress.

[0097] In summary, this invention endows the core components with targeted delivery properties through specific covalent modification, and with the assistance of triple immune microenvironment regulation, it constructs a three-dimensional synergistic system of immune remodeling, targeted anti-inflammation and targeted anti-oxidation, thereby effectively improving skin damage caused by 2,4-dinitrochlorobenzene.

[0098] Experiment 3: Animal Experiment on Anti-swelling Effect Animals: SPF-grade healthy Kunming mice, half male and half female, weighing 18-22g, were purchased from Beijing Speford Biotechnology Co., Ltd. Before the formal start of the experiment, all mice underwent a 7-day environmental acclimatization period. During this period, the temperature of the housing was maintained at 25±2℃, the humidity was controlled at 55%-65%, and a 12-hour light-dark cycle was provided. Mice had free access to food and water during the acclimatization period.

[0099] Drug administration: Mice were divided into normal group, model group, positive control group, blank matrix group, Example 1-3 group, comparative example 3 group, and comparative example 6-7 group, with 10 mice in each group. Hair was removed from the back of the mice using an electric hair shaver, with a hair removal area of ​​2cm×3cm.

[0100] Normal group: No medication administered.

[0101] Model group: No medication administered.

[0102] Blank matrix group: 0.2g of blank matrix was applied to the hair removal area of ​​each animal and administered continuously for 7 days.

[0103] Positive control group: Aspirin 200 mg / kg was administered by gavage daily for 7 consecutive days.

[0104] Examples 1-3, Comparative Examples 3 and Comparative Examples 6-7: 0.2g / animal of medical antipyretic and antipruritic ointment was evenly applied to the hair removal area and administered continuously for 7 days.

[0105] Modeling: 35 minutes after the last administration, except for the normal group, 20 μl of xylene was applied to the left ear of mice in all other groups to induce acute swelling of the auricle. The right ear was left untreated as a self-control. The model was considered successful when the left ear of the mouse swelled within 40 minutes.

[0106] Test: Mice were sacrificed 40 minutes after modeling. Both ears were cut off along the auricular baseline. Circular ear pieces were taken from the same location on both ears using a punch and precisely weighed. The weight difference between the left and right ear pieces was used as the degree of ear swelling. The swelling degree of each group of mice was recorded. The experimental results are shown in Table 3. Figure 2 As shown.

[0107] Table 3

[0108] Table 3 shows the comparison of swelling degree in xylene-induced auricular swelling models in each group of mice. The results showed that the auricular swelling degree in the model group was significantly higher than that in the normal group. Compared with the model group, the swelling degree in Examples 1-3 and each comparative group was reduced to varying degrees. Among them, the swelling degree in Example 2 was the lowest, and the swelling degree in Examples 1-3 was significantly lower than that in each comparative group. This indicates that the medical antipyretic and antipruritic ointment of the present invention has significant anti-inflammatory and swelling-reducing effects. The synergistic effect of hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex, modified polysaccharide and immune adjuvant is crucial to the swelling-reducing effect. Removing the key drug-loaded complex or omitting the drug-loaded modification treatment will lead to a significant reduction in the swelling-reducing effect.

[0109] Analysis of the comparative data showed that the swelling degree in Comparative Example 3 (with MPLA removed) was significantly increased to 4.35 ± 0.68, suggesting that in the multiple-dose intervention regimen of this invention, MPLA may induce local immune tolerance or regulatory immune cell recruitment through sustained low-level TLR4 activation, thereby reducing the tissue's sensitivity to inflammatory signals upon subsequent acute xylene stimulation. Simultaneously, MPLA, as a potent immune adjuvant, may also pre-establish a protective anti-inflammatory microenvironment by promoting the secretion of anti-inflammatory cytokines such as IL-10 by keratinocytes and local immune cells. The absence of this component alone resulted in the loss of the protective pretreatment effect, leading to a significant exacerbation of the acute swelling response.

[0110] The swelling values ​​of Comparative Example 6 (complete removal of curcumin complex) and Comparative Example 7 (physical mixing of equal amounts of free curcumin and blank chitosan) were as high as 5.60±1.05 and 5.15±0.98, respectively, significantly higher than those of Example 2 (2.68±0.32). This reveals the core advantage of the nano-drug delivery system of the present invention: hydroxypropyltrimethylammonium chloride chitosan, as a polycationic carrier, utilizes the electrostatic interaction between the positively charged groups on its molecular chain and the phenolic hydroxyl groups of curcumin, as well as the hydrophobic interaction and hydrogen bond synergy between the hydrophobic skeleton and the aromatic ring of curcumin, to achieve efficient encapsulation and stable dispersion of curcumin, allowing it to exist in an amorphous rather than crystalline state in the complex. This drug delivery system utilizes the positive charge characteristics of the carrier to enhance the electrostatic adsorption to the negatively charged inflammatory cell membrane, significantly overcoming the inherent defects of low solubility, easy crystallization, and poor transdermal permeability of free curcumin. This achieves efficient cross-barrier delivery and local long-term retention of curcumin at the lesion site, thereby precisely inhibiting cyclooxygenase-2 (COX-2) and nuclear factor. ( The signaling pathway was significantly reduced, as it significantly decreased the release of inflammatory mediators such as prostaglandin E2 (PGE2). In contrast, the physical mixing method in Comparative Example 7 failed to improve the crystallization state of curcumin or achieve carrier-mediated cellular uptake and transdermal absorption, resulting in a significant reduction in its anti-inflammatory and anti-swelling efficacy.

[0111] In summary, this invention achieves highly efficient blocking of xylene-induced acute inflammatory responses and rapid elimination of tissue edema by combining pretreatment-induced immunomodulation with the synergistic transmembrane delivery of a nanomedicine delivery system. Comparative data, from the opposite perspective, verify the indispensability of this synergistic design in the acute anti-edema efficacy.

[0112] Experiment 4: Animal Experiment on Antipruritic Efficacy Animals: SPF-grade male C57BL / 6 mice, weighing 20-22g, were purchased from Beijing Spefol Biotechnology Co., Ltd. Before the experiment officially began, all mice underwent a 7-day environmental acclimatization period. During this period, the temperature of the rearing environment was maintained at 25±2℃, the humidity at 55%-65%, and a 12-hour light-dark cycle was provided.

[0113] Two days before the experiment, all hair on the back of the neck of the mice was shaved off, with the hair removal area being approximately 2cm × 3cm, in preparation for subsequent drug administration and modeling procedures.

[0114] Drug administration: Mice were randomly divided into normal group, model group, positive control group, blank matrix group, Example 1-3 group, comparative example 1 group and comparative example 4-5 group, with 10 mice in each group. Hair was removed from the back of the mice using an electric hair shaver, with a hair removal area of ​​2cm×3cm.

[0115] Normal group: No medication administered.

[0116] Model group: No medication administered.

[0117] Positive control group: Diphenhydramine 10 mg / kg was injected intraperitoneally 30 minutes before modeling.

[0118] Examples 1-3, Comparative Example 1, and Comparative Example 4-5: 0.2g / animal of medical antipyretic and antipruritic ointment was evenly applied to the hair removal area and administered continuously for 4 days at 10:00 AM daily.

[0119] Blank matrix group: Apply 0.2g of blank matrix to the hair removal area per animal, administer for 4 consecutive days, at 10 am daily.

[0120] Modeling: At 11 a.m. on day 4 of the experiment, except for the normal group, all other groups of mice were injected intradermally with 500 μg of histamine in the right shoulder to construct a histamine-induced mouse acute pruritus model.

[0121] Testing: Mice were immediately placed in a quiet, transparent observation cage after modeling. The number of scratches was observed and recorded within 30 minutes. The criteria for judging scratching behavior were: the action of raising the hind paw and repeatedly scratching the modeled area was counted as a valid scratch; a single complete scratch was counted as one scratch. Licking, biting, forelimb rubbing, and other unrelated limb movements were not included in the count. The experimental results are shown in Table 4 and... Figure 3 As shown.

[0122] Table 4

[0123] Table 4 shows the comparison of scratching frequency in the histamine-induced acute pruritus model in each group of mice. The results showed that the scratching frequency in the model group was significantly higher than that in the normal group. Compared with the model group, the scratching frequency in Examples 1-3 and each comparative group was reduced to varying degrees, with Example 2 showing the fewest scratches. The scratching frequency in Examples 1-3 was significantly lower than that in each comparative group. This indicates that the medical antipyretic and antipruritic ointment of this invention has a significant antipruritic and soothing effect. Furthermore, the synergistic effect of glycyrrhizic acid-modified β-glucan, immune adjuvants, and peppermint oil is crucial to the antipruritic effect. Removing key active ingredients or omitting the modification treatment would significantly weaken the antipruritic efficacy.

[0124] The comparative data show that the removal of poly(I:C) (Comparative Example 1) led to a significant increase in the number of scratches. The mechanism is that poly(I:C), as a TLR3 agonist, gradually induced the expression of local type I interferon and anti-inflammatory cytokines during four consecutive days of administration, establishing an inhibitory immune microenvironment. When subsequently challenged with histamine, this microenvironment effectively weakened the sensitivity of sensory nerve endings to pruritogens and inhibited the cascade amplification of neurogenic inflammation. The absence of this component prevents the establishment of this protective pre-regulatory state, resulting in the inability to effectively block pruritus signal transduction.

[0125] Meanwhile, compared to using unmodified β-glucan (Comparative Examples 4 and 5), glycyrrhizic acid-modified β-glucan, covalently grafted with hydrazide bonds, leverages the specific recognition of β-glucan with the Dectin-1 receptor on the surface of immune cells such as macrophages to directionally enrich covalently bound glycyrrhizic acid in inflammation-related immune cells, thereby overcoming the shortcomings of low transdermal efficiency and insufficient local concentration of free glycyrrhizic acid. This targeted delivery strategy allows glycyrrhizic acid to more effectively inhibit histamine release and regulate TRPV1 channel activity, thus enhancing the antipruritic effect. In contrast, unmodified β-glucan lacks direct antihistamine activity and cannot achieve targeted delivery of glycyrrhizic acid, and it does not possess the synergistic anti-inflammatory effect brought about by grafted glycyrrhizic acid, thus its antipruritic efficacy is significantly weakened. It is worth noting that although the number of scratches in Comparative Example 5 (without modified β-glucan) was slightly lower than that in Comparative Example 4 (with glycyrrhizic acid completely removed and modified β-glucan), both were much higher than those in Example 2. This indicates that although unmodified β-glucan has a certain physical barrier or weak immune activation effect, it lacks active anti-allergic activity, which further verifies the necessity of hydrazide covalent grafting modification.

[0126] In summary, this invention endows the core components with targeted drug delivery properties through specific covalent modification and, supplemented by immune microenvironment pre-regulation, constructs an antipruritic system with synergistic effects of immune intervention and targeted anti-allergy, achieving significant antipruritic and soothing efficacy beyond the superposition of single components. Comparative data, from the opposite perspective, verify the indispensability of this synergistic design in the suppression of acute pruritus.

[0127] The above-described embodiments are merely illustrative of specific implementations of the present invention, and while the descriptions are detailed, they should not be construed as limiting the scope of protection of the present invention. It should be noted that for those skilled in the art, any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention should be considered equivalent substitutions and are included within the scope of protection of the present invention.

Claims

1. A prescription for a medical antipyretic and antipruritic ointment, characterized in that, The prescription is expressed as a percentage by weight as follows: Functional active ingredients: glycyrrhizic acid modified β-glucan 0.5%-2.0%, organoselenium-grafted sodium hyaluronate 1.0%-3.0%, hydroxypropyltrimethylammonium chloride chitosan-supported curcumin complex 1.0%-3.0%, polyinosinic acid 0.05%-0.2%, CpG oligonucleotide 0.01%-0.05%, monophospholipid A 0.0005%-0.01%, peppermint oil 0.1%-0.5%; wherein the glycyrrhizic acid modified β-glucan is glycyrrhizic acid modified β-glucan grafted with hydrazide bonds; Matrix excipients: 0.5%-1.5% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 5.0%-12.0% glycerin, 2.0%-5.0% 1,2-pentanediol, 8.0%-15.0% white petrolatum, 2.0%-5.0% polyglycerol-3-methyl glucoside distearate, 2.0%-5.0% dimethicone, 5.0%-10.0% caprylic / capric triglyceride, 0.5%-1.0% phenoxyethanol; Purified water: Top up to 100%.

2. The prescription for a medical antipyretic and antipruritic ointment according to claim 1, characterized in that, The prescription is more specifically as follows: Functional active ingredients: glycyrrhizic acid modified β-glucan 1.5%-1.8%, organoselenium-grafted sodium hyaluronate 1.5%-2.0%, hydroxypropyltrimethylammonium chloride chitosan-supported curcumin complex 2.0%-2.5%, polyinosinic acid 0.1%-0.15%, CpG oligonucleotide 0.032%-0.05%, monophospholipid A 0.008%-0.01%, peppermint oil 0.2%-0.4%; Matrix excipients: 0.8%-1.2% hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, 7.1%-9.4% glycerin, 3.0%-4.0% 1,2-pentanediol, 10.0%-12.0% white petrolatum, 4.0%-4.5% polyglycerol-3-methyl glucoside distearate, 3.0%-4.0% dimethicone, 6.06%-7.09% caprylic / capric triglyceride, 0.7%-0.9% phenoxyethanol; Purified water: Top up to 100%.

3. The prescription for a medical antipyretic and antipruritic ointment according to claim 2, characterized in that, The preparation method of glycyrrhizic acid-modified β-glucan grafted with hydrazide bonds is as follows: Yeast β-glucan is dissolved in purified water to prepare a 6% w / v dispersion. After hydration by magnetic stirring and ultrasonic dispersion at 35℃, the mixture is cooled to 4℃ and the pH is adjusted to 3.0-5.

0. Sodium periodate (1%-2% of the dry weight of β-glucan) is added and reacted in the dark for 2-4 hours to oxidize and open the ring, introducing aldehyde groups. Ethylene glycol is added to quench the aldehyde groups, and the aldehyde content is measured. Then, adipate dihydrazide is added in equimolar amounts to the aldehyde groups, and the reaction is maintained at pH 5.0 for 4 hours to form hydrazone bonds. Sodium cyanoborohydride (1.5 times the molar amount of the aldehyde groups) is then added and reacted at 4℃ for 2 hours to reduce the hydrazone bonds to stable secondary amine bonds, retaining the free hydrazide group at the other end of the adipate dihydrazide. Glycyrrhizic acid is added separately at a molar ratio of 1.2:1 to the free hydrazide group, using 0.1... A mol / L sodium hydroxide solution was used as a dissolving agent. EDC and NHS were added in a 1:1 molar ratio, each in an amount 1.2 times that of glycyrrhizic acid. The pH was adjusted to 6.0, and the mixture was activated at room temperature for 30 min to generate NHS ester. The activated solution was slowly added dropwise to the above reaction solution containing free hydrazide groups. The pH was adjusted to 6.0-6.5, and the reaction was carried out at 37℃ for 12-24 h to complete the hydrazide bond coupling. During this period, the consumption of free hydrazide groups was monitored using the ninhydrin colorimetric method. The reaction solution was centrifuged or filtered to remove insoluble matter. The filtrate was placed in a dialysis bag with a molecular weight cutoff of 30,000-50,000 Da and dialyzed with purified water at 4℃ for 24-48 h, with the water changed 4-6 times. The dialysate was freeze-dried and then pulverized through a 120-mesh sieve to obtain hydrazide-grafted glycyrrhizic acid modified β-glucan.

4. The prescription for a medical antipyretic and antipruritic ointment according to claim 2, characterized in that, The preparation method of the hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex is as follows: hydroxypropyltrimethylammonium chloride chitosan is added to purified water to prepare a 2 mg / mL aqueous solution, and stirred until completely dissolved; 10 mg / mL curcumin ethanol solution is slowly added dropwise at a loading of 15%, and the mixture is sonicated at 250W for 30 min to allow the curcumin to be fully loaded and bound; the mixture is purified by centrifugation at 3000g for 10 min using a 100 kDa ultrafiltration centrifuge tube, and the retentate is collected, freeze-dried, and then pulverized and passed through an 80-mesh sieve to obtain the hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex.

5. The prescription for a medical antipyretic and antipruritic ointment according to claim 2, characterized in that, The preparation method of the organoselenium-grafted sodium hyaluronate is as follows: Prepare a 7% aqueous solution of pharmaceutical-grade sodium hyaluronate, adjust the pH to 4.5-5.5, add 0.8 times the molar amount of EDC and an equimolar amount of NHS to the carboxyl group of hyaluronate, and activate at room temperature for 20 min; add 0.8% of the dry weight of sodium hyaluronate with selenomethionine, adjust the pH to 7.2, and react at room temperature for 8 h to form a stable amide bond; purify the product by dialysis with an 8000-12000 Da dialysis bag for 24 h, vacuum concentrate, spray dry, and pulverize through a 100-mesh sieve to obtain organoselenium-grafted sodium hyaluronate.

6. The prescription for a medical antipyretic and antipruritic ointment according to claim 2, characterized in that, The CpG oligonucleotide is selected from one of CpG oligonucleotide 1018, CpG oligonucleotide 1826, CpG oligonucleotide 2002, and CpG oligonucleotide 2395.

7. A method for preparing a medical antipyretic and antipruritic ointment according to any one of claims 1-6, characterized in that, The specific preparation method of the medical antipyretic and antipruritic ointment includes the following steps: S1. Preparation of aqueous phase: Add 80% of the prescribed amount of purified water to the emulsification reactor and heat to 75℃; add 80% of the prescribed amount of glycerol and the full amount of the prescribed amount of 1,2-pentanediol, stir at 500-600r / min for 10-15min until completely dissolved, and keep warm for later use. S2. Oil phase preparation: Add the full prescription amount of white petrolatum, the full prescription amount of dimethicone, the full prescription amount of caprylic / capric triglyceride, and the full prescription amount of peppermint oil to the oil phase tank in sequence, and heat to 80°C to melt; then add the full prescription amount of polyglycerol-3-methylglucose distearate, and stir at 80°C for 12-18 minutes until the system melts and becomes transparent to obtain a stable oil phase. S3. Emulsification and homogenization: Under stirring at 900-1100 r / min, the oil phase is slowly added to the water phase, and the mixture is stirred for 5 minutes at a controlled temperature of 75-80℃ for initial mixing; it is then refined into a fine paste by high-pressure homogenization, and then cooled naturally with cooling water. S4. Low-Temperature Active Compounding: After the system is cooled to below 40℃, add the full amount of hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer and stir at 450-550 r / min for 15-20 min until fully swollen; then add the full amount of organoselenium-grafted sodium hyaluronate and the full amount of glycyrrhizic acid-modified β-glucan, and stir to disperse evenly; grind the full amount of hydroxypropyltrimethylammonium chloride chitosan-loaded curcumin complex and the remaining 20% ​​of the formula amount of glycerin into a paste, and add the paste to the system after it has cooled to below 35℃. Stir at 600 rpm for 15-20 min until no visible particles are visible; add the full amount of polyinosinic acid and the full amount of CpG oligonucleotide to the pre-prepared monophospholipid A premixed suspension, stir until uniform under light-protected conditions, and then slowly add the paste in a thin stream under light-protected conditions, stirring at 300-400 rpm for 15-20 min to mix evenly; the monophospholipid A premixed suspension is prepared by taking 20% ​​of the prescribed amount of purified water, heating to 40℃, adding the full amount of monophospholipid A, and dispersing thoroughly by ultrasonication. S5. Finished product preparation: Add the full prescription amount of phenoxyethanol and stir at 400-500 r / min for 4-6 min to mix thoroughly; after adjusting the pH with buffer solution and degassing under negative pressure, place it under constant temperature conditions for aging; finally, aseptically fill it in a clean area that meets the requirements of pharmaceutical GMP to obtain the finished product.

8. The method for preparing a medical antipyretic and antipruritic ointment according to claim 7, characterized in that: In step S3, the high-pressure homogenization pressure is 30 MPa, and the homogenization cycle is 4-6 times.

9. The method for preparing a medical antipyretic and antipruritic ointment according to claim 7, characterized in that: In step S5, the vacuum degree of negative pressure degassing is -0.07 to 0.09 MPa, the constant temperature curing temperature is 23 to 27°C, and the curing time is 24 to 36 hours.

10. The method for preparing a medical antipyretic and antipruritic ointment according to claim 7, characterized in that: In step S5, the pH of the system is adjusted to 5.1-5.9 using a lactate-arginine buffer solution.

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