Rabdosia rubescens-based novel antibacterial and anti-inflammatory external preparation and preparation method thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine preparations, in particular to a rabdosia rubescens-based novel antibacterial and anti-inflammatory external preparation and a preparation method thereof, the rabdosia rubescens-based novel antibacterial and anti-inflammatory external preparation is prepared from the following raw materials in parts by weight: 100-200g of rabdosia rubescens, 50-100g of honeysuckle, 50-100g of fructus forsythiae, 30-80g of radix sophorae flavescentis, 30-80g of fructus cnidii and 10-30g of borneol, and the weight ratio of rabdosia rubescens to honeysuckle and fructus forsythiae is 1: (0.8-1.2). According to the novel anti-bacterial and anti-inflammatory external preparation based on the rabdosia rubescens and the preparation method, the preparation takes the rabdosia rubescens as a main material and is compatible with the traditional Chinese medicine components such as the honeysuckle, the fructus forsythiae, the radix sophorae flavescentis, the fructus cnidii and the borneol, and effective components are fully extracted by a method of combining ethanol reflux extraction and water decoction extraction; the traditional Chinese medicine composition can be prepared into various dosage forms such as lotion, gel, ointment and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine preparations, and in particular to a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens and a preparation method thereof. Background Art

[0002] Rabdosia rubescens, also known as ice grass, is a perennial herb or subshrub, or small shrub, of the genus Rabdosia in the Lamiaceae family. It is a variety of the plant Cardamine spp. in the genus Rabdosia. It gets its name from the thin, butterfly-shaped ice flakes that form on its surface, each as thin as a cicada's wing and in various shapes. The entire plant is covered with silvery-white borneol. It has anti-inflammatory, analgesic, stomach-tonifying, and blood-activating properties.

[0003] Because of its anti-inflammatory and analgesic properties, Rubescens rubescens is a promising candidate as a primary ingredient for topical antibacterial and anti-inflammatory preparations. However, existing topical anti-inflammatory preparations suffer from the following drawbacks: 1. Poor ingredient extraction selectivity: For example, the conventional decoction method employed in the previously published patent CN104888058B yields only 52.3% of the oridonin A extraction rate, and the high-temperature degradation rate of matrine exceeds 28% (accelerated test at 60°C); 2. Insufficient dosage form uniformity: For example, the gel formulation described in the previously published patent CN113546934A suffers from carbomer agglomeration and exhibits transdermal transdermal rate fluctuations exceeding 20% ​​(Franz diffusion cell method); and 3. Poor industrial adaptability: Existing processes lack equipment parameters (such as pulverization mesh size and filter press pressure), making mass production of tanks larger than 500L difficult. Therefore, we propose a novel topical antibacterial and anti-inflammatory preparation based on Rubescens rubescens and its preparation method. Summary of the Invention

[0004] The main purpose of the present invention is to provide a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens and a preparation method thereof, which can effectively solve the problems in the background technology.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] A novel antibacterial and anti-inflammatory external preparation based on rubescens is prepared from the following raw materials in parts by weight: 100-200g of rubescens, 50-100g of honeysuckle, 50-100g of forsythia, 30-80g of sophora flavescens, 30-80g of cnidium monnieri, and 10-30g of borneol, wherein the weight ratio of rubescens to honeysuckle+forsythia is 1:(0.8-1.2).

[0007] Preferably, it is prepared from the following raw materials in parts by weight: 150-180g of Rubescens herb, 60-90g of honeysuckle, 60-90g of forsythia, 40-70g of sophora flavescens, 40-70g of cnidium monnieri, and 15-25g of borneol, and the content of Rubescensine A in the extract is ≥1.5%.

[0008] Preferably, the gel is prepared from the following raw materials in parts by weight: 160g of Rabdosia rubescens, 80g of honeysuckle, 80g of forsythia, 60g of sophora flavescens, 60g of cnidium monnieri, and 20g of borneol. When the gel is prepared, the content of carbomer 940 is 1-3%.

[0009] A method for preparing a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens, characterized by comprising the following preparation steps:

[0010] Step 1, graded pulverization: pulverize Rabdosia rubescens, honeysuckle and forsythia suspensa to 20-40 mesh to obtain powder A, and pulverize Sophora flavescens and Cnidium monnieri to 10-30 mesh to obtain powder B;

[0011] Step 2, enzymatic extraction: add 70-75% ethanol (8 times the amount) to powder A, soak at 25°C for 2 hours, reflux at 80°C ± 2°C for 3 times (each time for 2 hours), and concentrate to a relative density of 1.20-1.30 to obtain extract C;

[0012] Step 3, inclusion water extraction: add water + 0.1-0.3% β-cyclodextrin (molar ratio 1:1) to powder B, decoct three times, filter under plate and frame pressure (0.2-0.4 MPa), and concentrate to a relative density of 1.15-1.25 to obtain extract D;

[0013] Step 4, extract purification: Extract C is purified by macroporous resin, and extract D is flocculated and impurities are removed by adding chitosan;

[0014] Step 5, preparation of dosage form: extract C and extract D are mixed at a ratio of 1:1-1:1.5, borneol ethanol solution is added, and a gel / ointment is prepared after ultrasonic dispersion.

[0015] Preferably, the ethanol concentration in step 2 is 72%, 0.1% cellulase is added in the first extraction, and the extraction rate of Rubescensine A is ≥92%.

[0016] Preferably, in step 3, the β-cyclodextrin inclusion makes the thermal decomposition temperature of matrine ≥210° C., the plate and frame filter pressure of the filtrate 0.3 MPa, and the pore size of the filter cloth ≤10 μm.

[0017] Preferably, in step 4, the extract C is purified by AB-8 resin (diameter-to-height ratio 1:10), the purity of Rubescensine A is ≥3.0%, the amount of chitosan added to the extract D is 0.1%, and the centrifugation speed is 4000 rpm.

[0018] Preferably, when preparing the gel in step 5, Carbomer 940 adopts a gradient swelling method (4° C., 12 h), a homogenization rate of 2000 rpm, and a pH of 5.8-6.5.

[0019] Preferably, the quality control of the preparation includes: Rubescensine A ≥ 1.5% (preparation), transdermal rate (borneol) ≥ 25 μg / cm 2 / h(3h), and the microbial limit complies with the requirements of the 2025 edition of the Chinese Pharmacopoeia.

[0020] The specific drug mechanism is as follows:

[0021] Rubescens: It tastes bitter and sweet, is slightly cold in nature, and enters the lung, stomach, and liver meridians. It has the effects of clearing away heat and detoxifying, promoting blood circulation and relieving pain, and having antibacterial and anti-inflammatory effects. Its active ingredients, Rubescensine A and B, have inhibitory effects on a variety of pathogens and can inhibit inflammatory reactions.

[0022] Honeysuckle: sweet in taste, cold in nature, enters the lung, heart and stomach meridians, clears away heat and detoxifies, and dispels wind-heat. The ingredients it contains, such as chlorogenic acid and luteolin, have broad-spectrum antibacterial, anti-inflammatory and antiviral effects.

[0023] Forsythia: bitter in taste, slightly cold in nature, enters the lung, heart, and small intestine meridians, clears away heat and detoxifies, reduces swelling and disperses nodules. Ingredients such as forsythiaside and forsythiaside have inhibitory effects on Staphylococcus aureus and Escherichia coli, and can relieve symptoms of redness, swelling, heat and pain in the inflamed area.

[0024] Sophora flavescens: bitter in taste, cold in nature, enters the heart, liver, stomach, large intestine, and bladder meridians, clears away heat and dampness, kills insects and relieves itching. Ingredients such as matrine and oxymatrine have antibacterial, anti-inflammatory, and anti-allergic effects, and are effective for skin fungal and bacterial infections.

[0025] Cnidium monnieri: It tastes pungent and bitter, is warm in nature, and enters the kidney meridian. It can dry dampness and dispel wind, kill insects and relieve itching. Its volatile oil and coumarin components have inhibitory effects on Candida albicans, Trichomonas, etc., and are often used to treat vulvar eczema, skin itching, etc.

[0026] Borneol: It tastes pungent and bitter, is slightly cold in nature, and enters the heart, spleen, and lung meridians. It can invigorate the mind, clear away heat and relieve pain. It also has the effect of promoting transdermal absorption and can enhance the skin penetration of other drug ingredients. It also has a cooling and antipruritic effect, relieving discomfort in the affected area.

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

[0028] 1. In the present invention, Rabdosia rubescens is the main drug, which clears away heat and toxins, and has antibacterial and anti-inflammatory effects; honeysuckle and forsythia are the assistant drugs, which enhance the heat-clearing and detoxifying effects; sophora flavescens and cnidium monnieri are the adjuvant drugs, which dry dampness and kill insects, and relieve itching; borneol is the guiding drug, which guides the drug to be absorbed through the skin and has a cooling and analgesic effect. The combination of these drugs has the effects of antibacterial, anti-inflammatory, detumescence and antipruritic.

[0029] 2. In the present invention, ethanol reflux is used to extract the fat-soluble components (such as oridonin and chlorogenic acid) in Rubescens, honeysuckle, and Forsythia, and water is used to extract the water-soluble components (such as matrine and osthole) in Sophora flavescens and Cnidium monnieri, to ensure that the effective ingredients are fully dissolved and the efficacy of the preparation is improved; the addition of borneol not only enhances transdermal absorption, but also reduces drug irritation and improves medication safety.

[0030] 3. The present invention can be made into various dosage forms such as lotion, gel, ointment, etc. according to clinical needs. It is suitable for various diseases such as skin infection, eczema, acne, traumatic inflammation, etc. It is easy to use and has strong adaptability. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The present invention is a flow chart of the method for improving the color fastness of fabrics. DETAILED DESCRIPTION

[0032] In order to make the technical means, creative features, objectives and effects achieved by the present invention easier to understand, the present invention is further described below in conjunction with specific implementation methods.

[0033] The raw material formula table of the present invention is as follows:

[0034]

[0035]

[0036] Example 1

[0037] A novel antibacterial and anti-inflammatory external preparation based on rubescens is prepared from the following raw materials in parts by weight (kg): 160kg of rubescens, 80kg of honeysuckle, 80kg of forsythia, 60kg of sophora flavescens, 60kg of cnidium monnieri, 20kg of borneol, and 20kg of carbomer 940.

[0038] A method for preparing a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens, comprising the following preparation steps:

[0039] Step 1, graded pulverization: pulverize Rabdosia rubescens, honeysuckle and forsythia suspensa to 30 mesh to obtain powder A, and pulverize Sophora flavescens and Cnidium monnieri to 20 mesh to obtain powder B;

[0040] Step 2, enzymatic extraction: add 1400 L of 72% ethanol to powder A, soak at 25°C for 2 h, reflux at 80°C ± 2°C for 3 times (2 h each time), and concentrate to a relative density of 1.25 to obtain extract C (28.5 kg, 1.82% of hydroxybenzoate);

[0041] Step 3, inclusion water extraction: Powder B was added with water + 2.4 kg of 0.2% β-cyclodextrin, decocted three times, and concentrated to a relative density of 1.2 by plate and frame filter pressing (0.3 MPa) to obtain extract D (22.3 kg, matrine 2.15%);

[0042] Step 4, extract purification: Extract C was purified by macroporous resin (yield 85%), and extract D was flocculated and impurities were removed by adding chitosan (turbidity ↓82%);

[0043] Step 5, preparation of dosage form: extract C and extract D were mixed at a ratio of 1:1.25, borneol ethanol solution was added, and the mixture was ultrasonically dispersed to form a gel / ointment.

[0044] Example 2

[0045] A novel antibacterial and anti-inflammatory external preparation based on Rabdosia rubescens is prepared from the following raw materials in parts by weight (kg): 100kg of Rabdosia rubescens, 50kg of honeysuckle, 50kg of forsythia, 30kg of sophora flavescens, 30kg of cnidium monnieri, and 10kg of borneol.

[0046] A method for preparing a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens, comprising the following preparation steps:

[0047] Step 1, graded pulverization: pulverize Rabdosia rubescens, honeysuckle and forsythia suspensa to 20 mesh to obtain powder A, and pulverize Sophora flavescens and Cnidium monnieri to 10 mesh to obtain powder B;

[0048] Step 2, enzymatic extraction: add 1200 L of 70% ethanol to powder A, soak at 25°C for 2 h, reflux at 80°C ± 2°C for 3 times (2 h each time), and concentrate to a relative density of 1.20 to obtain extract C;

[0049] Step 3, inclusion water extraction: add water + 2 kg of 0.1% β-cyclodextrin to powder B, decoct three times, filter by plate and frame pressure (0.2 MPa), and concentrate to a relative density of 1.15 to obtain extract D;

[0050] Step 4, extract purification: Extract C is purified by macroporous resin, and extract D is flocculated and impurities are removed by adding chitosan;

[0051] Step 5, preparation of dosage form: extract C and extract D were mixed in a ratio of 1:1, borneol ethanol solution was added, and the mixture was ultrasonically dispersed to form a gel / ointment.

[0052] Example 3

[0053] A novel antibacterial and anti-inflammatory external preparation based on Rabdosia rubescens is prepared from the following raw materials in parts by weight (kg): 200kg of Rabdosia rubescens, 100kg of honeysuckle, 100kg of forsythia, 80kg of sophora flavescens, 80kg of cnidium monnieri, and 30kg of borneol.

[0054] A method for preparing a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens, comprising the following preparation steps:

[0055] Step 1, graded pulverization: pulverize Rabdosia rubescens, honeysuckle and forsythia suspensa to 40 mesh to obtain powder A, and pulverize Sophora flavescens and Cnidium monnieri to 30 mesh to obtain powder B;

[0056] Step 2, enzymatic extraction: add 1600 L of 72% ethanol to powder A, soak at 25°C for 2 h, reflux at 80°C ± 2°C for 3 times (each time for 2 h), and concentrate to a relative density of 1.30 to obtain extract C;

[0057] Step 3, inclusion water extraction: add water + 0.3% β-cyclodextrin 3kg to powder B, decoct 3 times, filter by plate and frame pressure (0.3MPa), and concentrate to a relative density of 1.25 to obtain extract D;

[0058] Step 4, extract purification: Extract C is purified by macroporous resin, and extract D is flocculated and impurities are removed by adding chitosan;

[0059] Step 5, preparation of dosage form: extract C and extract D were mixed at a ratio of 1:1.5, borneol ethanol solution was added, and the mixture was ultrasonically dispersed to form a gel / ointment.

[0060] Experimental Example 4

[0061] Antibacterial effect test: experimental strains

[0062] Staphylococcus aureus (ATCC25923), Escherichia coli (ATCC25922), Candida albicans (ATCC10231).

[0063] Experimental methods

[0064] The lotion prepared in Example 1 was diluted to the stock solution concentration using the agar diffusion method. 10 μl of each aliquot was dropped into an Oxford cup containing a bacterial plate and cultured at 37° C. for 24 hours (48 hours for fungal culture). The diameter of the inhibition zone was measured.

[0065] Experimental results

[0066]

[0067] in conclusion

[0068] The preparation of the present invention has significant inhibitory effects on the three strains, indicating that it has broad-spectrum antibacterial activity.

[0069] Toxicology experiments

[0070] Twenty healthy mice were randomly divided into two groups (10 mice in each experimental group and control group). The experimental group was smeared with the gel of Example 2 on the back skin twice a day for 7 consecutive days; the control group was smeared with an equal amount of matrix gel. The skin reactions and systemic conditions of the mice were observed, and skin tissue was taken for pathological examination after the experiment. The results showed that there were no irritation reactions such as redness, swelling, and ulcers in the experimental group, and no abnormalities were found in histological observation; the skin condition of the control group was normal. This shows that the preparation of the present invention is highly safe and has no obvious toxic side effects.

[0071] Clinical Application

[0072] Eighty patients with skin infections were randomly divided into a treatment group (40 patients) who received the lotion of the present invention and a control group (40 patients who received a commercially available antibiotic ointment). The treatment group received the lotion two to three times daily for a seven-day course of treatment. Results showed an overall effective rate of 92.5% in the treatment group and 80.0% in the control group. The treatment group experienced significantly shorter redness and swelling resolution and pain relief than the control group (P < 0.05). No drug resistance or serious adverse reactions were observed in the treatment group.

[0073] In summary, the present invention discloses a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens and a preparation method thereof. The preparation is mainly composed of Rubescens rubescens and is combined with traditional Chinese medicine ingredients such as honeysuckle, forsythia, sophora flavescens, cnidium monnieri, and borneol. The effective ingredients are fully extracted by a method combining ethanol reflux extraction with water decoction extraction to prepare a variety of dosage forms such as lotions, gels, and ointments. The preparation of the present invention has the advantages of broad-spectrum antibacterial, significant anti-inflammatory, high safety, and good transdermal absorption. It is suitable for bacterial infections, inflammatory reactions, and other diseases of the skin and mucous membranes. The preparation has a simple preparation process and is suitable for industrial production.

[0074] The basic principles, main features, and advantages of the present invention are shown and described above. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions are merely illustrative of the principles of the present invention. Various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention. Such changes and modifications are intended to fall within the scope of the present invention. The scope of protection claimed in the present invention is defined by the appended claims and their equivalents.

Claims

1. A novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens, characterized by: The invention is prepared from the following raw materials in parts by weight: 100-200g of Rabdosia rubescens, 50-100g of honeysuckle, 50-100g of forsythia, 30-80g of sophora flavescens, 30-80g of cnidium monnieri and 10-30g of borneol, wherein the weight ratio of Rabdosia rubescens to honeysuckle+forsythia is 1:(0.8-1.2).

2. The novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens according to claim 1, characterized in that: The extract is prepared from the following raw materials in parts by weight: 150-180g of rubescens, 60-90g of honeysuckle, 60-90g of forsythia, 40-70g of sophora flavescens, 40-70g of cnidium monnieri and 15-25g of borneol, wherein the content of rubescensine A in the extract is greater than or equal to 1.5%.

3. The novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens according to claim 1, characterized in that: The gel is prepared from the following raw materials in parts by weight: 160g of Rabdosia rubescens, 80g of honeysuckle, 80g of forsythia, 60g of sophora flavescens, 60g of cnidium monnieri and 20g of borneol. When the gel is prepared, the content of carbomer 940 is 1-3%.

4. A method for preparing a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens according to any one of claims 1 to 3, characterized in that: The method comprises the following preparation steps: Step 1, graded pulverization: pulverize Rabdosia rubescens, honeysuckle and forsythia suspensa to 20-40 mesh to obtain powder A, and pulverize Sophora flavescens and Cnidium monnieri to 10-30 mesh to obtain powder B; Step 2, enzymatic extraction: add 70-75% ethanol (8 times the amount) to powder A, soak at 25°C for 2 hours, reflux at 80°C ± 2°C for 3 times (each time for 2 hours), and concentrate to a relative density of 1.20-1.30 to obtain extract C; Step 3, inclusion water extraction: add water + 0.1-0.3% β-cyclodextrin (molar ratio 1:1) to powder B, decoct three times, filter under plate and frame pressure (0.2-0.4 MPa), and concentrate to a relative density of 1.15-1.25 to obtain extract D; Step 4, extract purification: Extract C is purified by macroporous resin, and extract D is flocculated and impurities are removed by adding chitosan; Step 5, preparation of dosage form: extract C and extract D are mixed at a ratio of 1:1-1:1.5, borneol ethanol solution is added, and a gel / ointment is prepared after ultrasonic dispersion.

5. The method for preparing a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens according to claim 4, characterized in that: In the step 2, the ethanol concentration is 72%, 0.1% cellulase is added for the first extraction, and the extraction rate of Rubescensine A is ≥92%.

6. The method for preparing a novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens according to claim 4, characterized in that: In step 3, the β-cyclodextrin inclusion makes the thermal decomposition temperature of matrine ≥210° C., the plate and frame filter pressure of the filtrate 0.3 MPa, and the pore size of the filter cloth ≤10 μm.

7. The novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens and its preparation method according to claim 4, characterized in that: In step 4, the extract C is purified by AB-8 resin (diameter-to-height ratio 1:10), the purity of Rubescensine A is ≥3.0%, the amount of chitosan added to the extract D is 0.1%, and the centrifugation speed is 4000 rpm.

8. The novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens and its preparation method according to claim 4, characterized in that: When preparing the gel in step 5, Carbomer 940 adopts a gradient swelling method (4° C., 12 h), a homogenization rate of 2000 rpm, and a pH of 5.8-6.

5.

9. The novel antibacterial and anti-inflammatory external preparation based on Rubescens rubescens and its preparation method according to claim 4, characterized in that: Preparation quality control includes: Rubescensine A ≥ 1.5% (preparation), transdermal rate (borneol) ≥ 25 μg / cm 2 / h(3h), and the microbial limit complies with the requirements of the 2025 edition of the Chinese Pharmacopoeia.

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