Compound external ointment for treating leukoplakia vulvae and preparation method thereof

Abstract

The invention relates to the technical field of external ointment, in particular to compound external ointment for treating leukoplakia vulvae and a preparation method of the compound external ointment. 0.03 part of tacrolimus; 2.5 parts of hypocrellin; 5 parts of a cortex phellodendri extract; 10 parts of hyaluronic acid; 3 parts of vitamin E; and 70-75 parts of a matrix. Inflammation is rapidly controlled through low-efficiency glucocorticoid, local immune abnormality is inhibited through the immunomodulator, tissue is repaired through traditional Chinese medicine components, and bacteria are resisted, so that a quick-acting and long-acting combined mode is formed, hormone side effects are neutralized by adding hyaluronic acid and vitamin E, and the feasibility of the treatment course is prolonged; the total effective rate of the combined medicine for avoiding vitamin E oxidation through low-temperature mixing is remarkably higher than that of a traditional scheme, the occurrence rate of skin atrophy is reduced to 4% and far lower than 22% of that of the traditional scheme, and meanwhile, the staged mixing process ensures that the retention rate of active ingredients is larger than 95% and is remarkably superior to that of the traditional process.

Description

Technical Field

[0001] The invention relates to the technical field of external ointments, in particular to a composite external ointment for treating vulvar leukoplakia and a preparation method thereof. Background Art

[0002] At present, the mainstream treatments for vulvar leukoplakia include glucocorticoids (such as hydrocortisone) and immunosuppressants (such as tacrolimus), which can relieve itching and inflammation in the short term, but long-term use can easily cause side effects such as skin atrophy and dependence. Although Chinese medicine preparations (such as bamboo red fungus and phellodendron extract) are relatively safe, they have problems such as single ingredients, poor transdermal absorption, and slow efficacy. The existing technology attempts to compound hormones with traditional Chinese medicine, but has not solved the defects such as unclear component synergistic mechanism and poor process stability.

[0003] Therefore, in general, long-term use of hormone drugs in the traditional mainstream treatment of vulvar leukoplakia leads to skin barrier damage, and Chinese herbal compound lacks immune regulation function and cannot block the progression of the disease. At the same time, high-temperature extraction or mixing processes lead to degradation of active ingredients.

[0004] Based on this, the present invention provides a composite external ointment for treating vulvar leukoplakia and a preparation method thereof to solve the above-mentioned technical problems. Summary of the invention

[0005] The purpose of the present invention is to provide a composite external ointment for treating vulvar leukoplakia and a preparation method thereof, so as to solve the problems mentioned in the background technology.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] The present invention provides a composite external ointment for treating vulvar leukoplakia, which is composed of the following raw materials in parts by weight:

[0008] 0.3 parts of hydrocortisone;

[0009] Tacrolimus 0.03 parts;

[0010] 2.5 parts of hypocrellin;

[0011] 5 parts of Phellodendron chinense extract;

[0012] 10 parts of hyaluronic acid;

[0013] Vitamin E 3 parts;

[0014] 70-75 parts of matrix.

[0015] Preferably, the hydrocortisone is used as the monarch drug and is prepared by microbial hydroxylation reaction, with pregnenolone as the starting material, in an Aspergillus niger fermentation system, under the conditions of pH 6.5-7.0 and temperature 28-32°C, a hydroxylation reaction is carried out for 72 hours, and after purification by silica gel column chromatography and crystallization and drying, a white crystalline powder is obtained with a purity of ≥98%.

[0016] Preferably, the tacrolimus or pimecrolimus is used as an adjuvant, prepared by a Streptomyces fermentation method, the culture medium contains glucose and soy peptone, and is fermented for 10 days at pH 7.0 and 25°C. After macroporous resin adsorption and high performance liquid chromatography purification, a white crystalline powder is obtained with a purity of ≥99%.

[0017] Preferably, the hypocrellin extract is used as an adjuvant and is heated by supercritical CO 2 Extraction method, using the fruiting body of Bamboo Red Pleurotus as raw material, under pressure of 25MPa, temperature of 45℃, CO 2 The mixture was extracted for 3 hours at a flow rate of 20 L / min, and after reduced pressure concentration and freeze drying, a brown-yellow powder was obtained, with a total triterpenoid content of ≥90%.

[0018] Preferably, the Phellodendron amurense extract is used as an adjuvant and is prepared by reflux extraction with an ethanol-water mixed solvent, using the dried bark of Phellodendron amurense as the raw material, extracting at a temperature of 80°C for 2 hours, and obtaining a light yellow powder after rotary evaporation concentration and spray drying, with a berberine content of ≥8%.

[0019] Preferably, the hyaluronic acid is used as a drug, prepared by the fermentation method of Streptococcus equi, fermented at pH 6.8 and temperature 37°C for 48 hours, and purified by ultrafiltration and ethanol precipitation to obtain a white fibrous solid with a molecular weight of 1.5×10 6 Da, endotoxin content <0.05EU / mg.

[0020] Preferably, the vitamin E is used as a drug, separated and prepared from wheat germ oil by molecular distillation, and purified by rectification at a temperature of 200° C. and a vacuum degree of 0.1 mbar. After being collected in a cold trap and packaged under nitrogen protection, a light yellow oily liquid is obtained, and the α-tocopherol content is ≥96%.

[0021] Preferably, the matrix is ​​one of vaseline or polyethylene glycol.

[0022] Based on the above-mentioned composite external ointment for treating vulvar leukoplakia, the present invention also proposes a preparation method of the composite external ointment for treating vulvar leukoplakia, comprising the following steps:

[0023] S1. Extraction of Chinese medicinal ingredients: Phellodendron chinense was crushed and extracted with an ethanol-water mixed solvent, concentrated and dried into powder, and the hypocrellin was filtered through a supercritical CO 2 Extraction method:

[0024] S2. Mix the drug matrix, heat the matrix to 60-70°C, add hydrocortisone, tacrolimus, hypocrellin and Phellodendron chinense extract in sequence, and stir until uniform;

[0025] S3. Add auxiliary ingredients: After cooling to 40°C, add hyaluronic acid and vitamin E, and homogenize;

[0026] S4. Sterilization and filling: After ultraviolet sterilization, pack into aluminum tubes and store away from light.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The present invention uses low-efficiency glucocorticoids to quickly control inflammation, immunomodulators to inhibit local immune abnormalities, and Chinese herbal ingredients to repair tissues and have antibacterial effects, forming a "fast-acting + long-acting" combination mode. Hyaluronic acid and vitamin E are added to neutralize hormone side effects and extend the feasibility of treatment. Low-temperature mixing avoids vitamin E oxidation. The total effective rate of combined medication is significantly higher than that of traditional regimens, and the incidence of skin atrophy is reduced to 4%, which is much lower than the 22% of traditional regimens. At the same time, the staged mixing process ensures that the active ingredient retention rate is >95%, which is significantly better than the traditional process. DETAILED DESCRIPTION

[0029] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] Example 1. The present invention proposes a composite external ointment for treating vulvar leukoplakia, which is composed of the following raw materials in parts by weight: 0.3 parts of hydrocortisone; 0.03 parts of tacrolimus; 2.5 parts of bamboo red fungus; 5 parts of phellodendron chinense extract; 10 parts of hyaluronic acid; 3 parts of vitamin E; and 70-75 parts of a matrix.

[0031] In this embodiment, it should be noted that hydrocortisone is prepared as the main drug by microbial hydroxylation reaction, using pregnenolone as the starting material, and undergoing hydroxylation reaction for 72 hours at pH 6.5-7.0 and temperature 28-32°C in an Aspergillus niger fermentation system. After purification by silica gel column chromatography and crystallization and drying, a white crystalline powder is obtained with a purity of ≥98%.

[0032] In this embodiment, it should also be noted that tacrolimus or pimecrolimus is used as an adjuvant drug and is prepared by a Streptomyces fermentation method. The culture medium contains glucose and soy peptone and is fermented for 10 days at pH 7.0 and 25°C. After adsorption by a macroporous resin and purification by high performance liquid chromatography, a white crystalline powder is obtained with a purity of ≥99%.

[0033] In this embodiment, it should be noted that the hypocrellin extract was used as an adjuvant and was purified by supercritical CO 2 Extraction method, using the fruiting body of Bamboo Red Pleurotus as raw material, under pressure of 25MPa, temperature of 45℃, CO 2 The mixture was extracted for 3 hours at a flow rate of 20 L / min, and after reduced pressure concentration and freeze drying, a brown-yellow powder was obtained, with a total triterpenoid content of ≥90%.

[0034] In this embodiment, it should also be noted that the Phellodendron chinense extract is used as an adjuvant and is prepared by reflux extraction with an ethanol-water mixed solvent. The dried bark of Phellodendron chinense is used as a raw material and extracted at a temperature of 80°C for 2 hours. After rotary evaporation concentration and spray drying, a light yellow powder is obtained, and the berberine content is ≥8%.

[0035] In this example, it should be noted that hyaluronic acid was used as a drug, prepared by the fermentation method of Streptococcus equi, fermented at pH 6.8 and temperature 37°C for 48 hours, and purified by ultrafiltration and ethanol precipitation to obtain a white fibrous solid with a molecular weight of 1.5×10 6 Da, endotoxin content <0.05EU / mg.

[0036] In this embodiment, it should be noted that vitamin E is used as an adjuvant, separated and prepared from wheat germ oil by molecular distillation, and purified by rectification at a temperature of 200° C. and a vacuum degree of 0.1 mbar. After being collected in a cold trap and packaged under nitrogen protection, a light yellow oily liquid is obtained, and the α-tocopherol content is ≥96%.

[0037] In this embodiment, it should also be noted that the matrix is ​​one of vaseline or polyethylene glycol.

[0038] Example 2, based on the composite external ointment for treating vulvar leukoplakia in Example 1, this example also provides a method for preparing the composite external ointment for treating vulvar leukoplakia, specifically, comprising the following steps:

[0039] S1. Extraction of Chinese medicinal ingredients: Phellodendron chinense was crushed and extracted with an ethanol-water mixed solvent, concentrated and dried into powder, and the hypocrellin was filtered through a supercritical CO 2 Extraction method:

[0040] S2. Mix the drug matrix, heat the matrix to 60-70°C, add hydrocortisone, tacrolimus, hypocrellin and Phellodendron chinense extract in sequence, and stir until uniform;

[0041] S3. Add auxiliary ingredients: After cooling to 40°C, add hyaluronic acid and vitamin E, and homogenize;

[0042] S4. Sterilization and filling: After ultraviolet sterilization, pack into aluminum tubes and store away from light.

[0043] Design experiments to verify the efficacy, safety and process stability of the above-mentioned composite external ointment for the treatment of vulvar leukoplakia:

[0044] (1) Verification of efficacy improvement, comparing the total effective rate of the compound ointment of the present invention with that of 0.1% hydrocortisone alone;

[0045] a. Grouping:

[0046] Experimental group (n=100): using the compound ointment of the present invention, twice a day, for 4 consecutive weeks;

[0047] Control group (n=100): using the traditional regimen (0.1% hydrocortisone ointment), the usage is the same as the experimental group.

[0048] b.Evaluation criteria:

[0049] Significantly effective: Itching and erythema completely disappeared, and skin texture returned to normal;

[0050] Effective: Symptoms alleviated by ≥50%;

[0051] Ineffective: Symptoms improved by less than 50% or worsened.

[0052] Total effective rate = (number of markedly effective + effective) cases / total number of cases × 100%.

[0053] c. Data statistics: Chi-square test (χ 2 ) were used to analyze the differences between the groups (α=0.05).

[0054] (2) Safety optimization and verification: the incidence of skin atrophy after long-term use (12 weeks) of the ointment of the present invention was evaluated. 200 patients were included and used the ointment of the present invention once a day for 12 consecutive weeks.

[0055] a. Monitoring indicators:

[0056] Skin atrophy: measured by ultrasound of skin thickness (baseline vs. 12 weeks);

[0057] Incidence rate = number of cases with atrophy / total number of cases × 100%.

[0058] b. Comparative data: cite historical data.

[0059] (3) Process stability verification, testing the effect of the staged mixing process on the retention rate of active ingredients;

[0060] a. Batch preparation: Produce 5 batches of ointment consecutively and record the content of key ingredients in each batch.

[0061] b. Parameter control:

[0062] Mixing temperature: 60-70℃ for hormone / immunomodulator stage, ≤40℃ for auxiliary material stage;

[0063] Sterilization conditions: UV 254nm, 20 minutes.

[0064] c. Detection method:

[0065] Hydrocortisone, tacrolimus, hypocrellin: HPLC method:

[0066] Retention rate = (actual content / theoretical content) × 100%.

[0067] After the test is completed, record the data, see Table 1:

[0068] Table 1 Efficacy, safety and process stability test data

[0069]

[0070] In summary, combined with Table 1, it can be seen that the total effective rate of the combined drug of the present invention is significantly higher than that of the traditional regimen, and the incidence of skin atrophy is reduced to 4%, which is much lower than the 22% of the traditional regimen. At the same time, the staged mixing process ensures that the active ingredient retention rate is greater than 95%, which is significantly better than the traditional process.

[0071] In the description of this specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0072] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to the specific implementation methods described. Obviously, many modifications and changes can be made according to the content of this specification. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present invention, so that those skilled in the art can understand and use the present invention well. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. A composite external ointment for treating vulvar leukoplakia, characterized in that: It is composed of the following raw materials in parts by weight: 0.3 parts of hydrocortisone; Tacrolimus 0.03 parts; 2.5 parts of hypocrellin; 5 parts of Phellodendron chinense extract; 10 parts of hyaluronic acid; Vitamin E 3 parts; 70-75 parts of matrix.

2. The composite external ointment for treating vulvar leukoplakia according to claim 1, characterized in that: The hydrocortisone is used as the main drug and is prepared through a microbial hydroxylation reaction. Pregnenolone is used as the starting material and is subjected to a hydroxylation reaction for 72 hours in an Aspergillus niger fermentation system at pH 6.5-7.0 and a temperature of 28-32° C. The hydrocortisone is purified by silica gel column chromatography and crystallized and dried to obtain a white crystalline powder with a purity of ≥98%.

3. The composite external ointment for treating vulvar leukoplakia according to claim 2, characterized in that: The tacrolimus or pimecrolimus is used as an adjuvant drug and is prepared by a streptomyces fermentation method. The culture medium contains glucose and soy peptone and is fermented for 10 days at pH 7.0 and 25° C. After macroporous resin adsorption and high performance liquid chromatography purification, a white crystalline powder is obtained with a purity of ≥99%.

4. The composite external ointment for treating vulvar leukoplakia according to claim 3, characterized in that: The bamboo red fungus extract is used as an adjuvant and is prepared by supercritical CO2 extraction. The bamboo red fungus fruiting body is used as the raw material and extracted for 3 hours under the conditions of pressure 25MPa, temperature 45°C, and CO2 flow rate 20L / min. After reduced pressure concentration and freeze-drying, a brown-yellow powder is obtained, and the total triterpenoid content is ≥90%.

5. The composite external ointment for treating vulvar leukoplakia according to claim 4, characterized in that: The Phellodendron amurense extract is used as an adjuvant and is prepared by reflux extraction with an ethanol-water mixed solvent. The dried bark of Phellodendron amurense is used as a raw material and extracted at a temperature of 80° C. for 2 hours. After rotary evaporation concentration and spray drying, a light yellow powder is obtained, and the berberine content is ≥8%.

6. The composite external ointment for treating vulvar leukoplakia according to claim 5, characterized in that: The hyaluronic acid is used as a drug, prepared by the fermentation method of Streptococcus equi, fermented at pH 6.8 and temperature 37°C for 48 hours, and purified by ultrafiltration and ethanol precipitation to obtain a white fibrous solid with a molecular weight of 1.5×10 6 Da, endotoxin content <0.05EU / mg.

7. The composite external ointment for treating vulvar leukoplakia according to claim 6, characterized in that: The vitamin E is used as a drug, separated and prepared from wheat germ oil by molecular distillation, and purified by rectification at a temperature of 200° C. and a vacuum degree of 0.1 mbar. After being collected by a cold trap and packaged under nitrogen protection, a light yellow oily liquid is obtained, and the α-tocopherol content is ≥96%.

8. The composite external ointment for treating vulvar leukoplakia according to claim 7, characterized in that: The matrix is ​​one of vaseline or polyethylene glycol.

9. The method for preparing the composite external ointment for treating vulvar leukoplakia according to any one of claims 1 to 8, characterized in that: The following steps are involved: S1. Extraction of Chinese medicinal ingredients: Phellodendron chinense was crushed and extracted with an ethanol-water mixed solvent, concentrated and dried into powder, and hypocrellin was extracted by supercritical CO2 extraction; S2. Mixing drug matrix: Heat the matrix to 60-70°C, add hydrocortisone, tacrolimus, hypocrellin and Phellodendron chinense extract in sequence, and stir until uniform; S3. Add auxiliary ingredients: After cooling to 40°C, add hyaluronic acid and vitamin E, and homogenize; S4. Sterilization and filling: After ultraviolet sterilization, pack into aluminum tubes and store away from light.

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