Allicin-containing pharmaceutical composition for treating eczema and preparation method of allicin-containing pharmaceutical composition

Abstract

The invention relates to an allicin-containing pharmaceutical composition for treating eczema and a preparation method of the allicin-containing pharmaceutical composition, and belongs to the technical field of pharmaceutical compositions. In order to solve the problem that an existing eczema treatment medicine is difficult to consider anti-inflammatory, antibacterial and skin barrier repair at the same time, the invention provides the allicin-containing pharmaceutical composition for treating eczema, and the allicin-containing pharmaceutical composition contains the following effective pharmaceutical ingredients: an allicin inclusion compound, ceramide, sophocarpidine, baicalin, forsythin, asiaticoside and glycyrrhizic acid. Through scientific compatibility of various active components, an antibacterial-anti-inflammatory-repair triple synergistic treatment system is constructed, and the treatment targets of rapid symptom control in the acute stage, continuous repair in the chronic stage and prevention of infection and recurrence in the whole process of eczema treatment are achieved. Aiming at the defect of strong irritation of the allicin, the pharmaceutical composition containing the allicin is prepared into an oil-in-water cream form, so that the problems of stability and irritation of active ingredients of the allicin are solved, and the treatment targets of high efficiency and low toxicity are realized.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical compositions, and particularly relates to a pharmaceutical composition containing allicin for treating eczema and a preparation method thereof. BACKGROUND

[0002] Eczema is a common chronic inflammatory skin disease, mainly manifested as dry skin, erythema, pruritus, desquamation, and severe exudation, scabbing and lichenification. Its pathogenesis is complex, involving genetics, immune disorders, skin barrier dysfunction and environmental factors. Eczema can be divided into various types, such as atopic dermatitis, contact dermatitis and seborrheic dermatitis. There is no cure for eczema at present, and the main treatment goal is to relieve symptoms, reduce recurrence and improve the quality of life of patients.

[0003] The current clinical treatment drugs for eczema mainly include the following categories: glucocorticoids, calcineurin inhibitors, antihistamines, biological substances or antibiotics / antifungal drugs. The existing drugs have obvious side effects after long-term use, and the curative effect of some patients is poor, and it is difficult to meet the multiple needs of anti-inflammatory, antibacterial and skin barrier repair.

[0004] Allicin is the main active ingredient in garlic (Allium sativum), which is produced by enzymatic decomposition of alliin. Allicin has broad-spectrum antibacterial, anti-inflammatory and immunoregulatory, antioxidant and wound healing effects. At present, the application of allicin in the medical field is mainly oral preparations and external preparations. Among them, oral preparations have low bioavailability, are easily destroyed by gastric acid, and may cause gastrointestinal irritation. External preparations have poor stability, allicin is easily oxidized and decomposed, and have strong irritation, which easily causes skin burning sensation, limiting its clinical application. SUMMARY

[0005] In order to solve the problem that the existing eczema treatment drugs are difficult to meet the multiple needs of anti-inflammatory, antibacterial and skin barrier repair, the present application provides a pharmaceutical composition containing allicin for treating eczema and a preparation method thereof.

[0006] The technical scheme of the present application:

[0007] A pharmaceutical composition containing allicin for treating eczema contains the following effective pharmaceutical ingredients in mass percentage: allicin inclusion complex 0.5-1.0%, ceramide 2.0-3.0%, sophoridine 1.0-2.0%, matrine 0.5-1.5%, forsythoside 0.5-1.5%, asiaticoside 0.2-0.5% and glycyrrhizic acid 0.3-0.8%.

[0008] Further, the pharmaceutical composition is an oil-in-water cream type, and further contains the following components in the mass percentage: jojoba oil 6.0-8.0%, squalane 3.0-5.0%, glycerol monostearate 1.5-2.0%, sodium hyaluronate glycerol dispersion 0.1-0.2%, polysorbate 80 1.0-1.5%, cetyl stearyl alcohol 2.0-3.0%, and EDTA-2Na 0.05-0.1%, and the balance is purified water.

[0009] A preparation method of a pharmaceutical composition containing allicin for treating eczema, comprising the following steps:

[0010] Step one, preparation of allicin inclusion complex: β-cyclodextrin and allicin are mixed in purified water according to the molar ratio, and after stirring for a certain time, the solid product is collected by filtration, and after freeze-drying, the allicin inclusion complex is obtained;

[0011] Step two, preparation of sodium hyaluronate glycerol dispersion: glycerol and sodium hyaluronate powder are mixed uniformly to obtain a gel-like semi-solid, and then purified water is added for swelling, and the mixture is ready for use;

[0012] Step three, preparation of oil phase: jojoba oil, squalane, ceramide, glycerol monostearate, and cetyl stearyl alcohol are mixed and stirred to completely melt under water bath heating condition, and then the mixture is ready for use after being kept warm;

[0013] Step four, preparation of water phase: the remaining purified water and EDTA-2Na are mixed, and after being heated, sodium hyaluronate glycerol dispersion, polysorbate 80, allicin inclusion complex, oxymatrine, baicalin, forsythoside, and asiaticoside are added in sequence, and then constant temperature stirring is performed until complete dissolution, and the mixture is ready for use after being kept warm;

[0014] Step five, preparation of emulsion system: the oil phase prepared in step three is slowly poured into the water phase prepared in step four under stirring condition, and after all the oil phase is poured into the water phase, stepwise temperature and speed change homogenization treatment is performed, and then the emulsion system is obtained;

[0015] Step six, cooling post-treatment: when the emulsion system obtained in step five is cooled to 40°C under stirring condition, glycyrrhizic acid is added, the pH value of the system is adjusted to 5.5-6.0, and then the system is continuously stirred and cooled, and after vacuum degassing, the system is filled, and then an oil-in-water cream is obtained.

[0016] Further, the molar ratio of β-cyclodextrin to allicin in step one is 3-4:1, the temperature of the purified water is 30-40°C, and the stirring time is 2-3h.

[0017] Further, the molecular weight of sodium hyaluronate in step two is 10kDa, the mass ratio of glycerol to sodium hyaluronate powder is 3:1, the amount of purified water added is 150-200 times the mass of the sodium hyaluronate powder, and the swelling time is 30min.

[0018] Furthermore, the water bath heating temperature in step three is 75±1℃, and the stirring time is 15~20min.

[0019] Furthermore, in step four, the heating temperature is 75±1℃, and the stirring time is 20~30min.

[0020] Furthermore, in step five, the stirring state is 500 rpm, and the first stage of the segmented temperature-speed homogenization is homogenization at 2000 rpm for 3 minutes at 75℃, followed by water bath cooling to 60℃ and homogenization at 3000 rpm for 5 minutes.

[0021] Furthermore, in step six, the stirring speed for cooling down to 40°C is 200-300 rpm, and the stirring speed for continuing to cool down is 100-150 rpm, and stirring is stopped when the temperature reaches 30°C.

[0022] Furthermore, the viscosity of the oil-in-water emulsion obtained in step six is ​​20,000~40,000 cP at 25°C.

[0023] The beneficial effects of this invention are:

[0024] The allicin-containing pharmaceutical composition provided by this invention constructs a triple synergistic treatment system of "antibacterial-anti-inflammatory-repair" through the scientific formulation of multiple active ingredients. In terms of anti-inflammatory effects, the allicin inclusion complex can rapidly inhibit acute redness, swelling, heat, and pain, reducing the frequency of scratching within 24 hours; baicalin and glycyrrhizic acid form a dual antipruritic mechanism, reducing chronic itching, while matrine enhances the antipruritic effect by inhibiting mast cell degranulation. Regarding antibacterial effects, allicin and matrine synergistically cover Staphylococcus aureus / Malassezia, increasing the bacterial clearance rate to 99.5%; forsythoside disrupts the biofilm structure, and asiaticoside inhibits bacterial adhesion, effectively preventing secondary infections. In terms of barrier repair, ceramides rebuild the stratum corneum structure, reducing transepidermal water loss by 45% within 72 hours; asiaticoside promotes collagen synthesis, accelerating wound healing; and sodium hyaluronate forms a moisturizing film, increasing skin hydration. The pharmaceutical formulation provided by this invention achieves the treatment goals of rapid symptom control in the acute phase of eczema, continuous repair in the chronic phase, and prevention of infection recurrence throughout the entire treatment process.

[0025] To address the issue of strong irritation from allicin, this invention formulates an oil-in-water emulsion dosage form of the allicin-containing pharmaceutical composition, thus resolving the problems of stability and irritation associated with the active ingredient of allicin. This invention utilizes β-cyclodextrin inclusion technology and an outer aqueous phase to form a protective barrier, reducing the contact between allicin and oxygen and improving its stability. Simultaneously, the emulsified particles control the allicin particle size to 50-100 nm, enhancing transdermal absorption efficiency. Furthermore, jojoba oil and squalane can form a lipid-like membrane structure, further reducing the direct irritation of allicin to the stratum corneum, achieving a highly effective and low-toxicity therapeutic goal. Detailed Implementation

[0026] The technical solution of the present invention will be further described below with reference to embodiments, but it is not limited thereto. Any modifications or equivalent substitutions to the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention should be covered within the protection scope of the present invention. In the following embodiments, the process equipment or apparatus not specifically specified are all conventional equipment or apparatus in the art. Unless otherwise specified, the raw materials used in the embodiments of the present invention are all commercially available; unless otherwise specified, the technical means used in the embodiments of the present invention are all conventional means well known to those skilled in the art.

[0027] Example 1

[0028] This embodiment provides a pharmaceutical composition containing allicin for treating eczema and its preparation method.

[0029] In this embodiment, the allicin-containing pharmaceutical composition for treating eczema consists of the following components in weight percentage: allicin inclusion complex 0.5%, ceramide 2.0%, matrine 0.5%, baicalin 1.0%, forsythoside 0.5%, asiaticoside 0.2%, glycyrrhizic acid 0.3%, jojoba oil 6.0%, squalane 3.0%, glyceryl monostearate 1.5%, sodium hyaluronate glyceryl dispersion 0.1%, polysorbate 80 1.0%, cetearyl alcohol 2.0%, EDTA-2Na 0.05%, and the balance being purified water.

[0030] In this embodiment, the allicin-containing pharmaceutical composition used to treat eczema is an oil-in-water emulsion cream, and the specific preparation method includes the following steps:

[0031] Step 1: Preparation of allicin inclusion complex:

[0032] β-cyclodextrin and allicin were dissolved in purified water at 30°C at a molar ratio of 3:1. After stirring for 2 hours, the solid product was collected by filtration and freeze-dried to obtain allicin inclusion complex.

[0033] Step 2: Preparation of sodium hyaluronate-glycerol dispersion:

[0034] Glycerin and sodium hyaluronate powder with a molecular weight of 10 kDa were mixed evenly at a mass ratio of 3:1 to obtain a gel-like semi-solid. The gel was then swollen with 200 times the mass of purified water containing sodium hyaluronate powder for 30 minutes and set aside.

[0035] Step 3: Preparation of the oil phase:

[0036] Add jojoba oil, squalane, ceramide, glyceryl monostearate, and cetearyl alcohol to a beaker, heat in a water bath to 75°C, stir for 15 minutes until completely melted, and keep warm for later use.

[0037] Step 4: Prepare the aqueous phase:

[0038] Mix the remaining purified water and EDTA-2Na, heat to 75°C, and then add sodium hyaluronate glycerol dispersion, polysorbate 80, allicin inclusion complex, matrine, baicalin, forsythoside and asiaticoside in sequence. Stir at constant temperature for 20 minutes until completely dissolved.

[0039] Step 5: Prepare the emulsion system:

[0040] Slowly pour the oil phase prepared in step three into the aqueous phase prepared in step four, maintaining a stirring speed of 500 rpm during the pouring process. After all the oil phase is poured in, transfer it to a homogenizer and homogenize at 2000 rpm for 3 minutes. Then, cool the water bath to 60°C and homogenize at 3000 rpm for 5 minutes to ensure that the oil droplet size is ≤10 μm, which is consistent with the size of the skin hair follicle channel, thereby enhancing transdermal absorption and obtaining an emulsified system.

[0041] Step Six: Post-cooling treatment:

[0042] In step five, the emulsion system was stirred at 300 rpm and cooled to 40°C. Glycyrrhizic acid was added, and the pH of the system was adjusted to 6.0 with citric acid / sodium citrate buffer. Stirring was continued at 150 rpm and cooled to 30°C. Stirring was then stopped, and the mixture was vacuum degassed and filled to obtain an oil-in-water cream.

[0043] The water-in-oil cream obtained in this embodiment has a pH value of 6.0, a viscosity of 30,000 cP at 25°C, and no stratification during centrifugation at 3,000 rpm for 30 min.

[0044] Example 2

[0045] This embodiment provides a pharmaceutical composition containing allicin for treating eczema and its preparation method.

[0046] In this embodiment, the allicin-containing pharmaceutical composition for treating eczema consists of the following components in weight percentage: allicin inclusion complex 0.8%, ceramide 2.5%, matrine 1.0%, baicalin 1.5%, forsythoside 1.0%, asiaticoside 0.3%, glycyrrhizic acid 0.5%, jojoba oil 7.0%, squalane 4.0%, glyceryl monostearate 1.8%, sodium hyaluronate glyceryl dispersion 0.15%, polysorbate 80 1.2%, cetearyl alcohol 2.5%, EDTA-2Na 0.08%, and the balance being purified water.

[0047] In this embodiment, the allicin-containing pharmaceutical composition used to treat eczema is an oil-in-water emulsion cream, and the specific preparation method includes the following steps:

[0048] Step 1: Preparation of allicin inclusion complex:

[0049] β-cyclodextrin and allicin were dissolved in purified water at 40°C at a molar ratio of 4:1. After stirring for 3 hours, the solid product was collected by filtration and freeze-dried to obtain allicin inclusion complex.

[0050] Step 2: Preparation of sodium hyaluronate-glycerol dispersion:

[0051] Glycerin and sodium hyaluronate powder with a molecular weight of 10 kDa were mixed evenly at a mass ratio of 3:1 to obtain a gel-like semi-solid. The gel was then swollen with 150 times the mass of purified water containing sodium hyaluronate powder for 30 minutes and set aside.

[0052] Step 3: Preparation of the oil phase:

[0053] Add jojoba oil, squalane, ceramide, glyceryl monostearate, and cetearyl alcohol to a beaker, heat in a water bath to 75°C, stir for 20 minutes until completely melted, and keep warm for later use.

[0054] Step 4: Prepare the aqueous phase:

[0055] Mix the remaining purified water and EDTA-2Na, heat to 75°C, and then add sodium hyaluronate glycerol dispersion, polysorbate 80, allicin inclusion complex, matrine, baicalin, forsythoside and asiaticoside in sequence. Stir at constant temperature for 25 minutes until completely dissolved.

[0056] Step 5: Prepare the emulsion system:

[0057] Slowly pour the oil phase prepared in step three into the aqueous phase prepared in step four, maintaining a stirring speed of 500 rpm during the pouring process. After all the oil phase is poured in, transfer it to a homogenizer and homogenize at 2000 rpm for 3 min. Then, cool it to 60°C in a water bath and homogenize it at 3000 rpm for 5 min to ensure that the oil droplet size is ≤10 μm, thus obtaining the emulsified system.

[0058] Step Six: Post-cooling treatment:

[0059] In step 5, the emulsion system was stirred at 200 rpm and cooled to 40°C. Glycyrrhizic acid was added, and the pH of the system was adjusted to 5.5 with citric acid / sodium citrate buffer. The system was then stirred at 100 rpm and cooled to 30°C. Stirring was stopped, and the mixture was vacuum degassed and then filled to obtain an oil-in-water cream.

[0060] The water-in-oil cream obtained in this embodiment has a pH value of 5.5, a viscosity of 20,000 cP at 25°C, and no stratification during centrifugation at 3,000 rpm for 30 min.

[0061] Example 3

[0062] This embodiment provides a pharmaceutical composition containing allicin for treating eczema and its preparation method.

[0063] In this embodiment, the allicin-containing pharmaceutical composition for treating eczema consists of the following components in weight percentage: allicin inclusion complex 1.0%, ceramide 3.0%, matrine 1.5%, baicalin 2.0%, forsythoside 1.5%, asiaticoside 0.5%, glycyrrhizic acid 0.8%, jojoba oil 8.0%, squalane 5.0%, glyceryl monostearate 2.0%, sodium hyaluronate glyceride dispersion 0.2%, polysorbate 80 1.5%, cetearyl alcohol 3.0%, EDTA-2Na 0.1%, and the balance being purified water.

[0064] In this embodiment, the allicin-containing pharmaceutical composition used to treat eczema is an oil-in-water emulsion cream, and the specific preparation method includes the following steps:

[0065] Step 1: Preparation of allicin inclusion complex:

[0066] β-cyclodextrin and allicin were dissolved in purified water at 30°C at a molar ratio of 3:1. After stirring for 2 hours, the solid product was collected by filtration and freeze-dried to obtain allicin inclusion complex.

[0067] Step 2: Preparation of sodium hyaluronate-glycerol dispersion:

[0068] Glycerin and sodium hyaluronate powder with a molecular weight of 10 kDa were mixed evenly at a mass ratio of 3:1 to obtain a gel-like semi-solid. The gel was then swollen with 200 times the mass of purified water containing sodium hyaluronate powder for 30 minutes and set aside.

[0069] Step 3: Preparation of the oil phase:

[0070] Add jojoba oil, squalane, ceramide, glyceryl monostearate, and cetearyl alcohol to a beaker, heat in a water bath to 75°C, stir for 15 minutes until completely melted, and keep warm for later use.

[0071] Step 4: Prepare the aqueous phase:

[0072] Mix the remaining purified water and EDTA-2Na, heat to 75°C, and then add sodium hyaluronate glycerol dispersion, polysorbate 80, allicin inclusion complex, matrine, baicalin, forsythoside and asiaticoside in sequence. Stir at constant temperature for 20 minutes until completely dissolved.

[0073] Step 5: Prepare the emulsion system:

[0074] Slowly pour the oil phase prepared in step three into the aqueous phase prepared in step four, maintaining a stirring speed of 500 rpm during the pouring process. After all the oil phase is poured in, transfer it to a homogenizer and homogenize at 2000 rpm for 3 min. Then, cool it to 60°C in a water bath and homogenize it at 3000 rpm for 5 min to ensure that the oil droplet size is ≤10 μm, thus obtaining the emulsified system.

[0075] Step Six: Post-cooling treatment:

[0076] In step five, the emulsion system was stirred at 300 rpm and cooled to 40°C. Glycyrrhizic acid was added, and the pH of the system was adjusted to 6.0 with citric acid / sodium citrate buffer. Stirring was continued at 150 rpm and cooled to 30°C. Stirring was then stopped, and the mixture was vacuum degassed and filled to obtain an oil-in-water cream.

[0077] The water-in-oil cream obtained in this embodiment has a pH value of 6.0, a viscosity of 40,000 cP at 25°C, and no stratification during centrifugation at 3,000 rpm for 30 min.

[0078] Comparative Example 1

[0079] This comparative example provides a base cream and its preparation method.

[0080] In this comparative example, the base cream is composed of the following components in weight percentage: jojoba oil 8.0%, squalane 5.0%, glyceryl monostearate 2.0%, sodium hyaluronate glyceryl dispersion 0.2%, polysorbate 80 1.5%, cetearyl alcohol 3.0%, EDTA-2Na 0.1%, and the balance being purified water.

[0081] The specific preparation method of this comparative base cream includes the following steps:

[0082] Step 1: Preparation of sodium hyaluronate-glycerol dispersion:

[0083] Glycerin and sodium hyaluronate powder with a molecular weight of 10 kDa were mixed evenly at a mass ratio of 3:1 to obtain a gel-like semi-solid. The gel was then swollen with 200 times the mass of purified water containing sodium hyaluronate powder for 30 minutes and set aside.

[0084] Step 2, Preparation of the oil phase:

[0085] Add jojoba oil, squalane, glyceryl monostearate, and cetearyl alcohol to a beaker, heat in a water bath to 75°C, stir for 15 minutes until completely melted, and keep warm for later use.

[0086] Step 3: Prepare the aqueous phase:

[0087] Mix the remaining purified water and EDTA-2Na, heat to 75°C, then add sodium hyaluronate glycerol dispersion and polysorbate 80 in sequence, and stir at constant temperature for 20 minutes until completely dissolved;

[0088] Step 4: Prepare the emulsion system:

[0089] Slowly pour the oil phase prepared in step 2 into the aqueous phase prepared in step 3, maintaining a stirring speed of 500 rpm during the pouring process. After all the oil phase is poured in, transfer it to a homogenizer and homogenize at 2000 rpm for 3 min. Then, cool the water bath to 60℃ and homogenize at 3000 rpm for 5 min to ensure that the oil droplet size is ≤10 μm, thus obtaining the emulsified system.

[0090] Step 5, Post-cooling treatment:

[0091] The emulsion system obtained in step 4 was stirred at 300 rpm and cooled to 40°C. The pH of the system was adjusted to 6.0 with citric acid / sodium citrate buffer. The system was then stirred at 150 rpm and cooled to 30°C. Stirring was stopped, and the system was degassed under vacuum and then filled to obtain an oil-in-water cream.

[0092] Comparative Example 2

[0093] This comparative example provides a cream formulation containing only allicin and its preparation method.

[0094] The allicin-containing cream formulation in this comparative example consists of the following components by weight percentage: allicin inclusion complex 1.0%, jojoba oil 8.0%, squalane 5.0%, glyceryl monostearate 2.0%, sodium hyaluronate glyceryl dispersion 0.2%, polysorbate 80 1.5%, cetearyl alcohol 3.0%, EDTA-2Na 0.1%, and the balance being purified water.

[0095] The specific preparation method of the cream formulation containing only allicin in this comparative example includes the following steps:

[0096] Step 1: Preparation of allicin inclusion complex:

[0097] β-cyclodextrin and allicin were dissolved in purified water at 30°C at a molar ratio of 3:1. After stirring for 2 hours, the solid product was collected by filtration and freeze-dried to obtain allicin inclusion complex.

[0098] Step 2: Preparation of sodium hyaluronate glycerol dispersion:

[0099] Glycerin and sodium hyaluronate powder with a molecular weight of 10 kDa were mixed evenly at a mass ratio of 3:1 to obtain a gel-like semi-solid. The gel was then swollen with 200 times the mass of purified water containing sodium hyaluronate powder for 30 minutes and set aside.

[0100] Step 3: Prepare the oil phase:

[0101] Add jojoba oil, squalane, glyceryl monostearate, and cetearyl alcohol to a beaker, heat in a water bath to 75°C, stir for 15 minutes until completely melted, and keep warm for later use.

[0102] Step 4: Prepare the aqueous phase:

[0103] Mix the remaining purified water and EDTA-2Na, heat to 75°C, and then add sodium hyaluronate glycerol dispersion, polysorbate 80, and allicin inclusion complex in sequence. Stir at constant temperature for 20 minutes until completely dissolved.

[0104] Step 5: Prepare the emulsion system:

[0105] Slowly pour the oil phase prepared in step 3 into the aqueous phase prepared in step 4, maintaining a stirring speed of 500 rpm during the pouring process. After all the oil phase is poured in, transfer it to a homogenizer and homogenize at 2000 rpm for 3 min. Then, cool the water bath to 60℃ and homogenize at 3000 rpm for 5 min to ensure that the oil droplet size is ≤10 μm, thus obtaining the emulsified system.

[0106] Step 6, Post-cooling treatment:

[0107] The emulsion system obtained in step 5 was stirred at 300 rpm and cooled to 40°C. The pH of the system was adjusted to 6.0 with citric acid / sodium citrate buffer. The system was then stirred at 150 rpm and cooled to 30°C. Stirring was stopped, and the mixture was degassed under vacuum and then filled to obtain an oil-in-water cream.

[0108] Comparative Example 3

[0109] This comparative example provides a cream formulation containing only matrine and its preparation method.

[0110] The cream formulation containing only matrine in this comparative example is composed of the following components in weight percentage: matrine 1.5%, jojoba oil 8.0%, squalane 5.0%, glyceryl monostearate 2.0%, sodium hyaluronate glyceryl dispersion 0.2%, polysorbate 80 1.5%, cetearyl alcohol 3.0%, EDTA-2Na 0.1%, and the balance being purified water.

[0111] The specific preparation method of the cream formulation containing only matrine in this comparative example includes the following steps:

[0112] Step 1: Preparation of sodium hyaluronate-glycerol dispersion:

[0113] Glycerin and sodium hyaluronate powder with a molecular weight of 10 kDa were mixed evenly at a mass ratio of 3:1 to obtain a gel-like semi-solid. The gel was then swollen with 200 times the mass of purified water containing sodium hyaluronate powder for 30 minutes and set aside.

[0114] Step 2, Preparation of the oil phase:

[0115] Add jojoba oil, squalane, glyceryl monostearate, and cetearyl alcohol to a beaker, heat in a water bath to 75°C, stir for 15 minutes until completely melted, and keep warm for later use.

[0116] Step 3: Prepare the aqueous phase:

[0117] Mix the remaining purified water and EDTA-2Na, heat to 75°C, and then add sodium hyaluronate glycerol dispersion, polysorbate 80, and matrine in sequence. Stir at constant temperature for 20 minutes until completely dissolved.

[0118] Step 4: Prepare the emulsion system:

[0119] Slowly pour the oil phase prepared in step 2 into the aqueous phase prepared in step 3, maintaining a stirring speed of 500 rpm during the pouring process. After all the oil phase is poured in, transfer it to a homogenizer and homogenize at 2000 rpm for 3 min. Then, cool the water bath to 60℃ and homogenize at 3000 rpm for 5 min to ensure that the oil droplet size is ≤10 μm, thus obtaining the emulsified system.

[0120] Step 5, Post-cooling treatment:

[0121] When the emulsion system obtained in step 4 is cooled to 40°C by stirring at 300 rpm, the pH of the system is adjusted to 6.0 with citric acid / sodium citrate buffer. Stirring is continued at 150 rpm until the temperature drops to 30°C. Stirring is then stopped, and the mixture is vacuum degassed and filled to obtain an oil-in-water cream.

[0122] Comparative Example 4

[0123] This comparative example provides a common cream-based pharmaceutical composition for the treatment of eczema and its preparation method.

[0124] The eczema treatment drug composition in the conventional cream formulation of this comparative example consists of the following components in weight percentage: free allicin 1.0%, ceramide 3.0%, matrine 1.5%, baicalin 2.0%, forsythoside 1.5%, asiaticoside 0.5%, glycyrrhizic acid 0.8%, jojoba oil 8.0%, squalane 5.0%, glyceryl monostearate 2.0%, glycerin 5.0%, cetearyl alcohol 3.0%, white petrolatum 10.0%, EDTA-2Na 0.1%, and the balance being purified water.

[0125] The specific preparation method of the ordinary cream formulation in this comparative example includes the following steps:

[0126] Step 1: Preparation of the oil phase:

[0127] Add jojoba oil, squalane, ceramide, glyceryl monostearate, cetearyl alcohol, and white petrolatum to a beaker, heat in a water bath to 75°C, stir for 15 minutes until completely melted, and keep warm for later use.

[0128] Step 2, Prepare the aqueous phase:

[0129] Mix the remaining purified water, EDTA-2Na and glycerol, heat to 75°C, and then add matrine, baicalin, forsythoside and asiaticoside in sequence. Stir at constant temperature for 20 minutes until completely dissolved.

[0130] Step 3: Prepare the emulsion system:

[0131] Slowly pour the oil phase prepared in step 1 into the aqueous phase prepared in step 2, maintaining a stirring speed of 500 rpm during the pouring process. After all the oil phase is poured in, transfer it to a homogenizer and homogenize it at 2000 rpm for 3 minutes to form a uniform paste.

[0132] Step 4: Add active ingredients:

[0133] The paste obtained in step 3 is stirred at 300 rpm and cooled to 40°C. Free allicin and glycyrrhizic acid are added, and stirring is continued until completely dispersed.

[0134] Step 5: Adjust pH and cool:

[0135] The pH of the system was adjusted to 6.0 using citric acid / sodium citrate buffer solution. The mixture was then stirred at 150 rpm and cooled to 30°C before stirring was stopped. After vacuum degassing, the mixture was filled into containers to obtain a regular cream.

[0136] Experimental Example

[0137] This experiment verified the synergistic effect of the drugs through in vitro cell experiments and mouse eczema model experiments. Combined with the influence of the water-in-oil formulation on the stability of the components, the anti-inflammatory, antibacterial and barrier repair functions of the allicin-containing pharmaceutical composition for treating eczema provided by this invention were systematically evaluated.

[0138] I. In vitro cell experiments

[0139] 1. Cell model:

[0140] (1) RAW264.7 macrophages: used to detect the release of TNF-α and IL-6 inflammatory factors, seeding concentration: 2×10 5 cells / mL, 100 μL per well in a 96-well plate, final density 2 × 10⁻⁶. 4 cells / pores;

[0141] (2) HaCaT keratinocytes: used to assess the expression of barrier repair-related FLG and LOR proteins, seeding concentration: 1.5 × 10⁻⁶ 5 cells / mL, 2 mL per well in a 6-well plate, final density 3×10⁻⁶ 5 cells / pores.

[0142] 2. Grouping and drug treatment methods:

[0143] (1) Blank control group: 100 μL (96-well plate) or 2 mL (6-well plate) of DMEM medium containing 1% DMSO.

[0144] (2) LPS induction group: 100 μL of LPS (96-well plate) or 2 mL (6-well plate), final concentration 1 μg / mL;

[0145] (3) Allicin group: 100 μL (96-well plate) or 2 mL (6-well plate) of allicin dilution, final concentration 10 μM;

[0146] (4) Baicalin group: 100 μL (96-well plate) or 2 mL (6-well plate) of baicalin dilution solution, final concentration 20 μM;

[0147] (5) Glycyrrhizic acid group: 100 μL (96-well plate) or 2 mL (6-well plate) of glycyrrhizic acid dilution solution, final concentration 5 μM;

[0148] (6) Dosage form group: The oil-in-water cream preparation prepared in Example 3 was diluted with DMEM culture medium at a volume ratio of 1:100 and added to 100 μL (96-well plate) or 2 mL (6-well plate).

[0149] Treatment method: RAW264.7 macrophages were seeded into 96-well plates, and HaCaT keratinocytes were seeded into 6-well plates. After 24 hours of culture, the cells adhered to the plates. The original culture medium was discarded, and the cells were pretreated with drug-containing culture medium or diluted cream for 2 hours. Except for the blank control group, the other groups were added with LPS (1 μg / mL) and cultured for another 24 hours.

[0150] 3. Detection indicators:

[0151] (1) Collect cell supernatant from 96-well plates and detect the concentrations of TNF-α and IL-6 by ELISA;

[0152] (2) Collect cells from 6-well plates and extract total protein using RIPA lysis buffer. Western blotting was used to detect the expression levels of FLG and LOR. The results are shown in Table 1.

[0153] Table 1

[0154]

[0155] As shown in Table 1, the water-in-oil cream formulation prepared in this invention exhibits a 2-3 fold increase in the inhibition rate of TNF-α / IL-6 compared to the single-drug group, indicating that baicalin blocks the MAPK pathway, allicin inhibits NF-κB nuclear translocation, and glycyrrhizic acid downregulates AP-1 activity, forming a multi-target inhibition. The ceramide and asiaticoside in the water-in-oil cream formulation significantly promote FLG / LOR expression compared to the single-drug group.

[0156] II. Mouse Eczema Model Experiment

[0157] 1. Animal Model Construction Methods

[0158] BALB / c mice were shaved on their backs (2cm x 2cm), and eczema was induced by applying 0.2% MC903 (calcineurin inhibitor) for 7 days. On day 8, they were vaccinated with 10... 8 CFU Staphylococcus aureus ATCC25923 suspension.

[0159] 2. Grouping and Dosing Methods

[0160] (1) Model control group (n=10): The matrix cream prepared in Comparative Example 1 was applied;

[0161] (2) Allicin cream group (n=10): The cream containing only allicin prepared in Comparative Example 2 was applied;

[0162] (3) Matrine cream group (n=10): The cream containing only matrine prepared in Comparative Example 3 was applied;

[0163] (4) Ordinary cream group (n=10): The eczema treatment drug composition in ordinary cream form prepared in Comparative Example 4 was applied;

[0164] (5) Water-in-oil cream group (n=10): Apply the eczema treatment drug composition in the form of water-in-oil cream prepared in Example 3;

[0165] Administration method: Apply 20 μL / animal daily for 14 consecutive days.

[0166] 3. Detection indicators:

[0167] (1) Clinical score: Record the number of erythema, edema and scratching each day.

[0168] Erythema score (0-3 points):

[0169] 0 points: No erythema or skin consistent with normal skin;

[0170] 1 point: Slight pinkish spots with indistinct borders;

[0171] 2 points: Obvious red patches with clear borders;

[0172] 3 points: Dark red or purplish-red spots, accompanied by elevated skin temperature.

[0173] Edema score (0-3 points):

[0174] 0 points: Smooth skin without bumps;

[0175] 1 point: Slight localized bulge, which slowly rebounds after acupressure;

[0176] 2 points: Obvious edema, indentation lasts for more than 2 seconds after finger pressure;

[0177] 3 points: Severe swelling with a feeling of tension in the skin, no pitting when pressed.

[0178] Quantification of scratching behavior (times / 10 minutes):

[0179] Video surveillance was used to record the events, with 10-minute intervals as the observation unit.

[0180] 0 times: 0 points, 1-5 times: 1 point, 6-10 times: 2 points, ≥11 times: 3 points.

[0181] Comprehensive score calculation: Daily total score = erythema score + edema score + scratching score; the results are shown in Table 2.

[0182] Table 2

[0183]

[0184] As shown in Table 2, the score of the oil-in-water cream group on day 14 was 76.9% lower than that of the model control group, and significantly better than that of the allicin cream group and the matrine cream group. However, the scores of the oil-in-water cream group and the ordinary cream group were similar, with no statistical difference.

[0185] (2) Bacterial load: On day 14, skin lesion tissue was homogenized, spread on MH agar plates and counted. The results are shown in Table 3.

[0186] Table 3

[0187]

[0188] As shown in Table 3, the bacterial load in both the regular cream group and the oil-in-water cream group decreased to 10. 3 CFU / g level, compared to the model group (10 6 The CFU / g ratio decreased by three orders of magnitude, and was significantly better than the single-drug group. This indicates that allicin, matrine, and forsythoside in the ordinary cream group and the oil-in-water cream group form a synergistic antibacterial effect, covering different survival stages of Staphylococcus aureus, and have a good antibacterial effect in preventing secondary infections. Moreover, the antibacterial effect mainly depends on the drug compatibility, and the dosage form has little impact.

[0189] (3) Tewameter® TM300 detection of transepidermal water loss (TEWL): Mice were placed in the detection environment for 30 minutes to adapt. The probe was preheated for 10 minutes before use, disinfected with alcohol and then dried. Transepidermal water loss in the back model area was detected. The higher the value, the more severe the barrier damage. The results are shown in Table 4.

[0190] Table 4

[0191]

[0192] As shown in Table 4, the water-in-oil formulation outperforms the ordinary cream group in reducing TEWL and increasing the surface moisture content of the interwoven skin, and is more conducive to the repair of the skin barrier.

Claims

1. A pharmaceutical composition containing allicin for treating eczema, characterized in that, It contains the following active pharmaceutical ingredients in the following percentages by weight: allicin inclusion complex 0.5-1.0%, ceramide 2.0-3.0%, matrine 0.5-1.5%, baicalin 1.0-2.0%, forsythoside 0.5-1.5%, asiaticoside 0.2-0.5%, and glycyrrhizic acid 0.3-0.8%.

2. The allicin-containing pharmaceutical composition for treating eczema according to claim 1, characterized in that, The pharmaceutical composition is an oil-in-water cream formulation, and also contains the following components in weight percentage: jojoba oil 6.0-8.0%, squalane 3.0-5.0%, glyceryl monostearate 1.5-2.0%, sodium hyaluronate glyceryl dispersion 0.1-0.2%, polysorbate 80 1.0-1.5%, cetearyl alcohol 2.0-3.0%, and EDTA-2Na 0.05-0.1%, with the balance being purified water.

3. A method for preparing a pharmaceutical composition containing allicin for treating eczema as described in claim 2, characterized in that, Includes the following steps: Step 1: Preparation of allicin inclusion complex: β-cyclodextrin and allicin were mixed in a molar ratio and dissolved in purified water. After stirring for a certain period of time, the solid product was collected by filtration and freeze-dried to obtain allicin inclusion complex. Step 2: Preparation of sodium hyaluronate-glycerol dispersion: Glycerol and sodium hyaluronate powder are mixed evenly to obtain a gel-like semi-solid, which is then swollen with purified water and set aside for later use; Step 3: Preparation of the oil phase: Mix jojoba oil, squalane, ceramide, glyceryl monostearate, and cetearyl alcohol, and stir until completely melted under water bath heating conditions, then keep warm for later use; Step 4: Preparation of aqueous phase: Mix the remaining purified water and EDTA-2Na, heat, and then add sodium hyaluronate glycerol dispersion, polysorbate 80, allicin inclusion complex, matrine, baicalin, forsythoside and asiaticoside in sequence. Stir at constant temperature until completely dissolved, and keep warm for later use. Step 5: Preparation of emulsion system: Under stirring, slowly pour the oil phase prepared in step 3 into the aqueous phase prepared in step 4. After all the phase is poured in, perform segmented temperature-speed homogenization to obtain the emulsion system. Step 6, Cooling and Post-processing: When the emulsion system obtained in Step 5 is cooled to 40°C under stirring, add glycyrrhizic acid to adjust the pH value of the system to 5.5~6.0, continue stirring and cooling, degas under vacuum and then fill to obtain an oil-in-water cream.

4. The method for preparing the allicin-containing pharmaceutical composition for treating eczema according to claim 3, characterized in that, In step one, the molar ratio of β-cyclodextrin to allicin is 3-4:1, the temperature of the purified water is 30-40℃, and the stirring time is 2-3 hours.

5. The method for preparing the allicin-containing pharmaceutical composition for treating eczema according to claim 3 or 4, characterized in that, The hyaluronic acid sodium in step two has a molecular weight of 10 kDa, the mass ratio of glycerol to hyaluronic acid sodium powder is 3:1, the amount of purified water added is 150 to 200 times the mass of hyaluronic acid sodium powder, and the swelling time is 30 min.

6. The method for preparing the allicin-containing pharmaceutical composition for treating eczema according to claim 5, characterized in that, The water bath heating temperature in step three is 75±1℃, and the stirring time is 15~20min.

7. The method for preparing the allicin-containing pharmaceutical composition for treating eczema according to claim 6, characterized in that, The heating temperature in step four is 75±1℃, and the stirring time is 20~30min.

8. The method for preparing the allicin-containing pharmaceutical composition for treating eczema according to claim 7, characterized in that, The stirring state described in step five is 500 rpm. The first stage of the segmented temperature-speed homogenization is to homogenize at 2000 rpm for 3 minutes at 75℃, and then cool down to 60℃ in a water bath and homogenize at 3000 rpm for 5 minutes.

9. The method for preparing the allicin-containing pharmaceutical composition for treating eczema according to claim 8, characterized in that, In step six, the stirring speed for cooling down to 40°C is 200-300 rpm, and the stirring speed for continuing to cool down is 100-150 rpm. Stirring is stopped when the temperature reaches 30°C.

10. The method for preparing the allicin-containing pharmaceutical composition for treating eczema according to claim 9, characterized in that, The viscosity of the oil-in-water emulsion obtained in step six is ​​20,000~40,000 cP at 25°C.

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