Colchicine dihydric alcohol plastid cataplasm and preparation method thereof

Abstract

The invention relates to the technical field of acute gout prevention and treatment, and discloses a colchicine dihydric alcohol plastid cataplasm which comprises colchicine dihydric alcohol plastid ingredients consisting of soybean phospholipid, cholesterol, colchicine, tween-80, ethanol and propylene glycol, the cataplasm further comprises cataplasm matrix ingredients composed of NP-700, glycerin, sodium carboxymethyl cellulose, diatomite, DL-tartaric acid, pure water and aluminum glycinate. According to the colchicine dihydric alcohol plastid cataplasm and the preparation method thereof, colchicine dihydric alcohol plastid, the A-phase solvent and the B-phase solvent are firstly prepared respectively, and then the colchicine dihydric alcohol plastid cataplasm is prepared, so that colchicine is prepared into a transdermal absorption preparation, the skin permeability is further improved, and the colchicine dihydric alcohol plastid cataplasm has a good application prospect. After local skin administration, the medicine does not pass through the gastrointestinal tract, can directly permeate into tissues from the skin surface, and reaches the affected part after being absorbed, so that the effect of quickly exerting the curative effect without causing untoward effects of the whole body is realized.

Description

Technical Field

[0001] The present invention relates to the technical field of acute gout prevention and treatment, and specifically to a colchicine diol plasmid cataplasm and a preparation method thereof. Background Art

[0002] Gout is a disease of acute or chronic inflammation and tissue damage caused by the precipitation of monosodium urate (MSU) crystals in bones, joints, kidneys, subcutaneous tissues and other various tissues. Colchicine is the earliest studied alkaloid and a classic drug for the treatment and prevention of acute gout. It also has certain anti-fibrosis and anti-tumor effects. According to the "Guidelines for the Diagnosis and Treatment of Hyperuricemia and Gout in China (2019)", the use of small doses (0.5-1.0 mg / d) of colchicine for 3 to 6 months can prevent gout attacks.

[0003] Today, after rigorous medical review, colchicine has been widely used in the clinical application. It can be used to treat familial Mediterranean fever, Behcet's disease, cardiovascular disease, cerebral stroke and some inflammatory diseases. Although colchicine is well tolerated at conventional doses, its therapeutic window is very narrow, and the effective dose and toxic dose are very close. In addition, due to the high toxicity of colchicine, it often produces certain side effects when used. Among them, the most common adverse reactions in clinical practice are nausea, vomiting, loss of appetite, diarrhea, weight loss, etc. Therefore, how to reduce adverse reactions while improving the efficacy has become the main research issue in the use of colchicine.

[0004] However, most of the current colchicine dosage forms on the market in China are oral dosage forms, which need to be digested and absorbed by the gastrointestinal tract, have a long effective time and are prone to cause adverse reactions in some people. Compared with ordinary oral administration, if colchicine is prepared into a transdermal absorption preparation, it can be administered topically without passing through the gastrointestinal tract. The drug can penetrate into the tissue through the skin surface, so that it can quickly exert its therapeutic effect without causing systemic adverse reactions. Therefore, in order to solve the above-mentioned technical problems, the present invention mainly prepares a diol plasmid that can improve the skin permeability of colchicine drugs, and prepares it into a colchicine diol plasmid patch, thereby accelerating the transdermal absorption of the drug and improving the treatment efficiency. Summary of the invention

[0005] 1. Technical issues to be resolved

[0006] In view of the deficiencies in the prior art, the present invention provides a colchicine diol plasmid patch and a preparation method thereof, which have the advantages of accelerating the transdermal absorption efficiency of the drug and being able to quickly exert therapeutic effects without causing systemic adverse reactions. It solves the problem in the above-mentioned background technology that the existing colchicine is mostly in oral dosage form, which takes a long time to treat through gastrointestinal digestion and absorption and is prone to adverse reactions due to drug side effects.

[0007] (II) Technical solution

[0008] In order to achieve the above-mentioned purpose of accelerating the efficiency of drug transdermal absorption and being able to quickly exert therapeutic effects without causing systemic adverse reactions, the present invention provides the following technical solutions: a colchicine diol plastosome cataplasm,

[0009] The invention is characterized in that it comprises a colchicine diol plasmid ingredient composed of soybean lecithin, cholesterol, colchicine, Tween-80, ethanol and propylene glycol and weighed and measured according to the prescription amount, and also comprises a papule base ingredient composed of NP-700, glycerol, sodium carboxymethyl cellulose, diatomaceous earth, DL-tartaric acid, pure water and glycolylated aluminum and weighed in sufficient amount.

[0010] Preferably, colchicine diol plastids are prepared according to the colchicine diol plastid ingredients, and the preparation steps are:

[0011] L1. Place the soybean lecithin, cholesterol, colchicine and Tween-80 weighed according to the prescription amount into a 40 mL brown vial;

[0012] L2. Then, add the ethanol and propylene glycol solutions according to the prescribed amount into a brown vial and sonicate for 20 minutes to fully dissolve them to obtain an alcohol phase solution;

[0013] L3. After adding the magnet to the alcohol phase solution, place it in a constant temperature magnetic stirrer, set the stirring temperature to 20°C-50°C and the stirring speed to 500-800r / min for uniform stirring;

[0014] L4. Add ceramide III powder to the alcohol phase solution at a concentration of 1 mg / mL and stir slowly to fully dissolve it;

[0015] L5. Measure the PBS buffer solution according to the prescription, use a syringe to draw it as the aqueous phase test solution, and slowly drip the PBS buffer solution into the alcohol phase solution while continuing to hydrate at a constant temperature for 20 minutes;

[0016] L6. After hydration, the mixture was filtered using a 0.22 μm filter membrane to prepare a colchicine diol plastid suspension having a blue opalescent luster and a light yellow color;

[0017] The matrix is ​​prepared according to the ingredients of the cataplasm, and the preparation steps are as follows:

[0018] T1. Take 8.12 g of glycerol in a beaker, and then evenly mix 1.27 g of NP-700 in glycerol to obtain a glycerol miscible solvent;

[0019] T2. 0.6 g of sodium carboxymethyl cellulose and 0.2 g of diatomaceous earth were added to the glycerol miscible solvent and placed in a magnetic stirrer and stirred at a speed of 200 r / min to prepare a phase A solvent;

[0020] T3. Take another beaker and weigh 1.56gDL-tartaric acid and 10g pure water and mix them evenly, then add 0.31g of aluminum glycolate and place the mixed solution in a magnetic stirrer;

[0021] T4. Set the stirring speed to 200 r / min, the stirring temperature to 40° C. and the stirring time to 20 min to obtain the phase B solvent after uniform mixing.

[0022] Preferably, the quality evaluation is performed according to the method of controlling the variables of the bile-egg ratio, soybean lecithin dosage, dosage, glycol ratio and alcohol phase dosage in the diol plastid ingredients, and the single factor screening of each formula dosage and process parameters is performed according to the quality evaluation method. The evaluation content includes three determination methods of particle size, Zeta potential and encapsulation efficiency. A comprehensive score is scored according to the evaluation content, and the scoring method is as follows:

[0023] Comprehensive score = particle size score × 25% + Zeta potential score × 25% + encapsulation efficiency score × 50%;

[0024] The evaluation samples were prepared according to the quality evaluation method, and the evaluation sample preparation method was as follows:

[0025] 1) Select a 25 mL volumetric flask and quantitatively measure 1 mL of the prepared colchicine diol plastid into the volumetric flask; 2) Add purified water to the colchicine diol plastid and dilute to the 25 mL mark, shake well and ultrasonicate for 20 minutes to fully disperse the aggregated diol plastids to obtain a diluted liquid sample.

[0026] Preferably, the particle size determination and evaluation steps are:

[0027] a1. Select several ultrasonically cleaned particle size test dishes, take an appropriate amount of diluent sample and transfer the diluent to each particle size test dish to form several first determination sample liquids;

[0028] a2. Measure three times in each first determination sample solution, take the average value of the obtained particle size data as the final result, and perform particle size scoring. The scoring method is as follows:

[0029]

[0030] The zeta potential determination and evaluation steps are as follows:

[0031] b1. Select several ultrasonically cleaned potential test dishes, take an appropriate amount of diluent in the diluent sample and transfer the diluent to each potential test dish to form several copies of the second measurement sample solution;

[0032] b2. Measure three times in each second determination sample solution, take the average value of the obtained Zeta potential data as the final result, and perform potential scoring, the scoring method is as follows:

[0033]

[0034] Preferably, the encapsulation efficiency determination and evaluation steps are:

[0035] c1. Select several 5mL volumetric flasks, measure 0.1mL of diol plasmid into each volumetric flask, and add methanol to mix until the 5mL mark;

[0036] c2. Each mixed sample was vortexed for 10 minutes and then filtered through a 0.22 μm microporous membrane. The total concentration of colchicine in each mixed sample was measured by high performance liquid chromatography (HPLC). 总 , each sample was measured three times;

[0037] c3. Select several dialysis bags with a molecular weight cutoff of 500, accurately measure 3 mL of colchicine diol plastids and transfer them to each dialysis bag, seal them with dialysis clips and place them in 150 mL of PBS solution;

[0038] c4. Take out 5 mL of solution from each dialysis bag every 2 hours and add the same volume of solution at the same temperature until the concentration of the solution remains unchanged after dialysis;

[0039] c5. Determine the concentration of colchicine in each dialyzed solution sample using a high performance liquid chromatograph 游 , each sample was measured three times;

[0040] Among them C 游 represents the amount of free drug, i.e., the drug not encapsulated by the diol plastids; C 总 is the total amount of drug in the colchicine diol plastid suspension; the encapsulation efficiency is expressed as The encapsulation rate scoring method is as follows:

[0041] Another technical problem to be solved by the present invention is to provide a method for preparing colchicine diol plastosome cataplasm, comprising the following preparation steps:

[0042] S1. Take an appropriate amount of colchicine diol plastid and add it to the prepared phase B solvent and stir to mix;

[0043] S2. Add the prepared phase A solvent to the mixed phase B solvent and mix again to obtain a mixed solution;

[0044] S3. Select several non-woven fabrics and evenly apply the obtained mixed solution on each non-woven fabric to prepare colchicine diol plasmid patch.

[0045] (III) Beneficial effects

[0046] Compared with the prior art, the present invention provides a colchicine diol plasmid cataplasm and a preparation method thereof, which has the following beneficial effects:

[0047] 1. The colchicine diol plasmid patch and its preparation method are as follows: colchicine diol plasmids and A phase solvent and B phase solvent are prepared respectively first, and then colchicine diol plasmid patch is prepared according to the colchicine diol plasmids and A phase solvent and B phase solvent, so that colchicine is prepared into a transdermal absorption preparation, and the skin permeability is further improved. After local skin administration, the drug can directly penetrate into the tissue from the skin surface without passing through the gastrointestinal tract, and reach the affected area after absorption, thereby achieving the effect of rapid therapeutic effect without causing systemic adverse reactions.

[0048] 2. The colchicine diol plasmid cataplasm and its preparation method have been effectively verified to have good safety after contact with the skin through multiple skin irritation tests. At the same time, combined with experimental data observations, it is known that the prepared colchicine diol plasmid cataplasm has a certain therapeutic effect on gout, and is superior to traditional oral tablets in terms of treatment methods and effects.

[0049] 3. The colchicine diol plasmid patch and its preparation method, by combining the prepared colchicine diol plasmid patch with non-woven fabric, are more convenient for patients to use in treatment in the form of a patch, avoiding the situation where the ointment is easily scratched when applied and dried, providing an isolated environment for the colchicine diol plasmid patch when it comes into contact with the skin, ensuring the purity of the drug during action, and avoiding external pollution that affects the efficacy. At the same time, a highly waterproof material layer structure can be added to the non-attached surface of the non-woven fabric, thereby further improving the environmental applicability of the colchicine diol plasmid patch. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 This is a schematic diagram of the experimental results of investigating the bile-egg ratio of the colchicine diol plastid ingredients of the present invention;

[0051] Figure 2 This is a schematic diagram of the experimental results of investigating the content of soybean lecithin in the colchicine diol plastidial ingredient of the present invention;

[0052] Figure 3 This is a schematic diagram of the experimental results of investigating the dosage of colchicine diol plastid ingredients of the present invention;

[0053] Figure 4 This is a schematic diagram of the experimental results of investigating the ethanol content in the diol of the colchicine diol plastid ingredient of the present invention;

[0054] Figure 5This is a schematic diagram of the experimental results of investigating the alcohol content of the colchicine diol plastid ingredients of the present invention;

[0055] Figure 6 This is a colchicine diol plastid sample diagram of the present invention;

[0056] Figure 7 It is a schematic diagram of the TEM measurement results in the colchicine diol plastid and the blank diol plastid of the present invention;

[0057] Figure 8 It is a schematic diagram of the particle size determination results in the colchicine diol plastid and the blank diol plastid of the present invention;

[0058] Fig. 9 This is a schematic diagram of the results of the in vitro release performance test of colchicine diol plastids of the present invention;

[0059] Fig.10 This is a schematic diagram of the experimental results of the colchicine diol plastid stability investigation of the present invention;

[0060] Fig.11 It is a schematic diagram of the in vitro transdermal test results of colchicine diol plasmids, alcohol solution and aqueous solution of the present invention;

[0061] Fig.12 This is a schematic diagram of the results of screening the amount of NP-700 in the matrix ingredients of the cataplasm of the present invention;

[0062] Fig.13 This is a schematic diagram of the screening results of the amount of aluminum glycolate in the base ingredients of the cataplasm of the present invention;

[0063] Fig.14 This is a schematic diagram of the results of screening the amount of tartaric acid in the matrix ingredients of the cataplasm of the present invention;

[0064] Fig.15 This is a schematic diagram of the results of screening the amount of sodium carboxymethyl cellulose in the base ingredients of the cataplasm of the present invention;

[0065] Fig.16 This is a schematic diagram of the results of screening the amount of glycerin in the base ingredients of the cataplasm of the present invention;

[0066] Fig.17 This is a schematic diagram of a non-woven fabric coated with colchicine diol plasmid patch of the present invention;

[0067] Fig.18 This is a schematic diagram of the skin irritation test results of a single administration of the colchicine diol plastosome cataplasm of the present invention;

[0068] Fig.19 This is a schematic diagram of the results of the skin irritation experiment of the colchicine diol plastosome cataplasm of the present invention after multiple administration;

[0069] Fig. 20 This is a schematic diagram of the experimental results of joint swelling in gout rats of the present invention. DETAILED DESCRIPTION

[0070] The following will be combined with the embodiments of the present invention and the accompanying drawings 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.

[0071] Embodiment 1:

[0072] A colchicine diol plastid patch comprises colchicine diol plastid ingredients composed of soybean lecithin, cholesterol, colchicine, Tween-80, ethanol and propylene glycol and weighed and measured according to the prescription amount. The colchicine diol plastid is prepared according to the colchicine diol plastid ingredients, and the preparation steps are as follows:

[0073] L1. Place the soybean lecithin, cholesterol, colchicine and Tween-80 weighed according to the prescription amount into a 40 mL brown vial;

[0074] L2. Then, add the ethanol and propylene glycol solutions according to the prescribed amount into a brown vial and sonicate for 20 minutes to fully dissolve them to obtain an alcohol phase solution;

[0075] L3. After adding the magnet to the alcohol phase solution, place it in a constant temperature magnetic stirrer, set the stirring temperature to 20°C-50°C and the stirring speed to 500-800r / min for uniform stirring;

[0076] L4. Add ceramide III powder to the alcohol phase solution at a concentration of 1 mg / mL and stir slowly to fully dissolve it;

[0077] L5. Measure the PBS buffer solution according to the prescription, use a syringe to draw it as the aqueous phase test solution, and slowly drip the PBS buffer solution into the alcohol phase solution while continuing to hydrate at a constant temperature for 20 minutes;

[0078] L6. After hydration, the mixture was filtered using a 0.22 μm filter membrane to prepare a colchicine diol plastid suspension with a blue opalescent luster and a light yellow color.

[0079] Specifically, in step L1, the weighed soybean lecithin, cholesterol, colchicine, Tween-80 and other ingredients are placed in a 40 mL brown vial. This is because colchicine is easily decomposed by light. In order to ensure the stability of colchicine in the entire preparation process, the brown vial is selected for light protection.

[0080] In step L6, the colchicine diol plastid suspension can be obtained by hydrating the alcohol phase solution at a constant temperature and then filtering it with a 0.22 μm filter membrane. In order to ensure that the prepared colchicine diol plastid has excellent stability, the colchicine diol plastid needs to be modified with ceramide III. The modification method is to add ceramide III powder with a mass concentration of 1 mg / mL to the alcohol phase solution in step L4, and mix it after dissolving.

[0081] Embodiment 2:

[0082] The bile-egg ratio, soybean lecithin dosage, dosage, glycol ratio and alcohol phase dosage in the diol plastid were investigated using the controlled variable method.

[0083] Specifically, the inspection method is as follows:

[0084] (I) Investigation of the bile-egg ratio

[0085] The dosage of other drug reagents and the preparation process in the prescription were kept unchanged. The quality of the colchicine binary pure plasmid was evaluated by changing the bile-egg ratio. The bile-egg ratio was set to 1:10, 2:10, 3:10, 4:10 and 5:10 as five conditional experimental groups. Three samples were prepared in parallel in each group, and the quality was evaluated according to the three determination methods of particle size, Zeta potential and encapsulation efficiency.

[0086] according to Figure 1 The experimental results in the results show that the optimal bile-egg ratio is 2:10. Different bile-egg ratios have little effect on the diol plasmid particle size and potential score, which tend to be flat, while the comprehensive score shows a downward trend after the bile-egg ratio reaches 2:10.

[0087] (II) Investigation of soybean lecithin content

[0088] The dosage of other drug reagents and the preparation process in the prescription were kept unchanged, and the quality of the obtained colchicine binary pure body was evaluated by changing the soybean lecithin content. The soybean lecithin content was set to 0.25%, 0.5%, 0.75%, 1% and 1.5% as five groups of conditional experimental groups, and three samples were prepared in parallel in each group. The quality was evaluated according to the three determination methods of particle size, Zeta potential and encapsulation efficiency.

[0089] according to Figure 2 The experimental results in the results show that the optimal soybean lecithin content is 0.75%. Different contents of soybean lecithin have a great influence on the particle size, potential and encapsulation efficiency of diol plastids. When the soybean lecithin content exceeds 0.75%, the comprehensive evaluation decreases significantly.

[0090] (III) Dosage Investigation

[0091] The dosage of other drug reagents and the preparation process in the prescription were kept unchanged, and the quality of the obtained colchicine binary pure plasmid was evaluated by changing the dosage. The dosage of binary alcohol plasmid was set to 2 mg, 4 mg, 6 mg, 8 mg and 10 mg as five conditional experimental groups, and three samples were prepared in parallel in each group. The quality was evaluated according to the three determination methods of particle size, Zeta potential and encapsulation efficiency.

[0092] according to Figure 3 The experimental results in the experiment showed that the optimal dosage was 6 mg. As the dosage increased, the potential increased and the encapsulation rate decreased. When the dosage was greater than 6 mg, the comprehensive score decreased significantly.

[0093] (IV) Investigation of ethanol content in diols

[0094] The dosage of other drug reagents and the preparation process in the prescription were kept fixed, and the quality of the obtained colchicine diol plasmid was evaluated by changing the dosage of ethanol. The ethanol content in the diols (ethanol and propylene glycol) was set to 40%, 50%, 60%, 70% and 80% respectively as five conditional experimental groups. Three samples were prepared in parallel in each group, and the quality was evaluated according to the three determination methods of particle size, Zeta potential and encapsulation efficiency.

[0095] according to Figure 4 The experimental results show that the highest quality score is when the ethanol content in the diol is 70%, and when the ethanol content in the diol is greater than 70%, the encapsulation efficiency score decreases significantly.

[0096] (V) Investigation of alcohol phase content

[0097] The dosage of other drug reagents and the preparation process in the prescription were kept fixed, and the quality of the obtained colchicine diol plasmid was evaluated by changing the hydration temperature. According to the preliminary experiment, the alcohol phase content was set to 30%, 40%, 50%, 60% and 70% as five conditional experimental groups, and three samples were prepared in parallel in each group. The quality was evaluated according to the three determination methods of particle size, Zeta potential and encapsulation efficiency.

[0098] according to Figure 5 The experimental results in the literature show that the optimal alcohol phase content is between 50% and 70%. As the amount of alcohol phase changes, the potential, particle size and encapsulation efficiency are significantly affected. When the alcohol phase content is greater than 60%, the comprehensive score decreases significantly.

[0099] Embodiment three:

[0100] The prepared colchicine diol plasmids were characterized and evaluated by appearance, TEM, particle size, stability, in vitro release and in vitro transdermal permeability. Before the characterization and evaluation, blank diol plasmids need to be prepared for comparative experiments with colchicine diol plasmids.

[0101] Specifically, the ingredients of the blank diol plasmid include soybean lecithin, cholesterol, Tween-80, ethanol and propylene glycol weighed and measured according to the prescription amount, and the preparation steps are as follows:

[0102] P1. Place the soybean lecithin, cholesterol and Tween-80 weighed according to the prescription amount into a 40 mL brown vial;

[0103] P2. Then, add ethanol and propylene glycol solution according to the prescribed amount into a brown vial and sonicate for 20 minutes to fully dissolve them to obtain an alcohol phase solution;

[0104] P3. After adding the magnetite to the alcohol phase solution, place it in a constant temperature magnetic stirrer, set the stirring temperature to 20°C-50°C and the stirring speed to 500-800r / min for uniform stirring;

[0105] P4. Add ceramide III powder to the alcohol phase solution at a concentration of 1 mg / mL and stir slowly to fully dissolve it;

[0106] P5. Measure the PBS buffer solution according to the prescription, use a syringe to draw it as the aqueous phase test solution, and slowly drip the PBS buffer solution into the alcohol phase solution while continuing to hydrate at a constant temperature for 20 minutes;

[0107] P6. After hydration, filter using a 0.22 μm filter membrane to prepare a blank diol plastid suspension.

[0108] (I) Appearance

[0109] According to the colchicine diol plastid ingredients, colchicine diol plastids can be prepared as follows: Figure 6 Shown is a light yellow suspension with a light blue opalescence.

[0110] 2. TEM

[0111] 101L of prepared colchicine diol plastids and blank diol plastids were respectively pipetted onto a copper grid, negatively stained with 2% phosphotungstic acid, and excess water was removed with filter paper and then dried under a drying lamp. After drying, the diol plastids were observed under a transmission electron microscope. The diol plastids were spherical vesicle-like structures, and there were no black particle shadows in the blank alcohol plastids (such as Figure 7As shown in the A part of the figure, the colchicine diol plastids and blank alcohol plastids have uniform particle sizes, smooth surfaces, and uniform distribution, while the colchicine diol plastids are evenly distributed with black particle shadows, which are colchicine drugs (such as Figure 7 (shown in part B).

[0112] (III) Particle size

[0113] The particle size of the diol plasmid was measured by Malvern nanoparticle size potentiometer. Two diol plasmids were prepared according to the optimal bile-egg ratio, soybean lecithin content, dosage, ethanol content in diol and alcohol phase content as the prescription. Three copies were prepared in parallel. The particle size of the blank alcohol plasmid was (102±0.97) nm, the particle size of the colchicine diol plasmid was (112±0.82) nm, and the particle size of the ceramide III-colchicine diol plasmid was (129.9±0.66) nm.

[0114] according to Figure 8 The measurement results show that its particle size is small and it is easy to be absorbed through the skin, thus achieving the purpose of drug efficacy quickly.

[0115] (IV) In vitro release performance of ethosomes

[0116] according to Fig. 9 The in vitro release curve of colchicine diol plasmids in PBS (pH=6.8) shows that the plasmids undergo a burst release in the first hour, with a drug release rate of about 60%. The drug release rate reaches a maximum and is close to 80% at 2 hours. Between 2 and 8 hours, the drug release rate is close to being flat, indicating that the drug-carrying plasmid vesicles are easy to release colchicine drugs in the in vitro simulated release experiment, and the colchicine drug has not deteriorated. If the plasmids are used in transdermal preparations, the skin can fully absorb the drug in a shorter time, and the effect is better.

[0117] (V) Stability investigation

[0118] Three groups of colchicine diol plasmids were prepared in parallel under the same conditions, and three samples were prepared in each group. The three groups of colchicine diol plasmids were placed at 4°C in the dark, 25°C in the dark, and 25°C in the light for 28 days, and the changes in their particle size and encapsulation efficiency were measured on the 0th day, the 4th day, the 8th day, the 12th day, the 16th day, the 20th day, the 24th day, and the 28th day.

[0119] according to Fig.10 The experimental results show that the particle size and encapsulation rate of colchicine diol plastids change little at 4℃ in the dark, indicating that it has good stability at 4℃ in the dark, which may be related to the reduced ethanol volatilization of diol plastids at 4℃ and the stability of soybean lecithin at low temperature.

[0120] However, under light and 25°C conditions, the particle size and encapsulation rate changed significantly, indicating that the diol plasmid was unstable under these conditions. Therefore, it was concluded that colchicine diol plasmid should be stored in a low-temperature, light-proof environment.

[0121] (VI) In vitro transdermal test

[0122] The experimental process is as follows: 1) After the rats are anesthetized with ether, the coarse hair and fine hair on the abdomen are removed, the depilated rat skin is cut with surgical scissors, and the rats are killed by cervical dislocation. The removed rat skin is firstly removed of excess subcutaneous tissue and fat, and then repeatedly rinsed with physiological saline, and the water is absorbed with filter paper, and then wrapped with tin foil and stored in a freezer at -20℃ for future use;

[0123] 2) Prepare Cer3-COL diol plasmids according to the optimal recipe, take a certain amount of the drug and dissolve it in the same volume of anhydrous ethanol to prepare colchicine alcohol solution, and then take a certain amount of the drug and dissolve it in the same volume of purified water to prepare colchicine aqueous solution, and ensure that the drug concentration (mg / mL) of the three test solution samples is the same;

[0124] 3) The in vitro transdermal performance of colchicine diol plasmids was studied using a modified Franze diffusion cell experiment. The volume of the receiving cell used in the experiment was 20 mL, and its effective diffusion area was 0.95 cm 2 , select physiological saline as the receiving liquid, take the prepared mouse skin, fix it between the receiving chamber and the supply chamber, with the stratum corneum facing the supply pool and the dermis facing the receiving pool, slowly inject physiological saline into the receiving pool, fill the receiving pool with physiological saline, carefully exhaust all bubbles, and make the mouse skin and physiological saline in close contact;

[0125] 4) Set the water bath temperature to (37±0.5)°C, add 4.0 mL of the three test solutions to the supply chamber, respectively, accurately sample 1.0 mL from the receiving pool at 2 h, 4 h, 6 h, 8 h, 10 h and 24 h, and simultaneously add an equal amount of isothermal fresh physiological saline. Filter the taken out receiving solution with a 0.22 μm filter membrane, and then determine and calculate its concentration according to the chromatographic conditions;

[0126] 5) Calculate the cumulative permeability per unit area Q at each time point n , with time t as the horizontal axis, the cumulative permeability per unit area is Q n Plot Q as the ordinate n-t The curve is regressed on t, and the calculation formula is:

[0127]

[0128] Where Q n is the cumulative permeability per unit area at the nth time point, C n is the concentration measured at the nth time point, Ci-1 is the concentration measured at the i-th (i=n-1) time point, V is the volume of the receiving pool, V i is the sampling volume, A is the effective receiving area of ​​the receiving pool;

[0129] According to the in vitro transdermal study experimental process, the measurement was repeated three times, and the drug concentrations of the three test solutions were set to be the same. The average cumulative transdermal amount of colchicine diol plasmid, alcohol solution and aqueous solution was obtained.

[0130] according to Fig.11 The experimental results showed that the cumulative transdermal doses per unit area in 24 hours were 28.91, 13.73 and 6.741 g·cm -2 The cumulative transdermal amount of colchicine diol plasmids was significantly higher than that of alcohol solution and aqueous solution, and was 2.1 times that of alcohol solution and 4 times that of aqueous solution, respectively. It can be seen that colchicine diol plasmids have good transdermal properties, making it easier for the drug to penetrate the skin.

[0131] Embodiment 4:

[0132] A colchicine diol plasmid cataplasm comprises cataplasm base ingredients consisting of NP-700, glycerol, sodium carboxymethyl cellulose, diatomaceous earth, DL-tartaric acid, pure water and aluminum glycolate weighed in sufficient amounts, and the preparation steps are as follows:

[0133] T1. Take 8.12 g of glycerol in a beaker, and then evenly mix 1.27 g of NP-700 in glycerol to obtain a glycerol miscible solvent;

[0134] T2. 0.6 g of sodium carboxymethyl cellulose and 0.2 g of diatomaceous earth were added to the glycerol miscible solvent and placed in a magnetic stirrer and stirred at a speed of 200 r / min to prepare a phase A solvent;

[0135] T3. Take another beaker and weigh 1.56gDL-tartaric acid and 10g pure water and mix them evenly, then add 0.31g of aluminum glycolate and place the mixed solution in a magnetic stirrer;

[0136] T4. Set the stirring speed to 200 r / min, the stirring temperature to 40° C. and the stirring time to 20 min to obtain the phase B solvent after uniform mixing.

[0137] Specifically, a screening test is conducted within a certain dosage range to screen a single factor for the cataplasm prescription, and then determine the optimal dosage of each component in the cataplasm matrix ingredients. The screening steps are as follows:

[0138] (1) NP-700 dosage screening

[0139] according to Fig.12The screening results show that the comprehensive score is higher when the dosage of NP-700 is in the range of 0.8g-1.4g. When the dosage exceeds 1.4g, the scores of all items of the matrix show a downward trend, resulting in a significant decrease in the comprehensive score of the matrix. During the preparation of the matrix, it was found that with the increase in the dosage of NP-700, excessive cross-linking with the cross-linking agent occurred, the cohesion was too large, the viscosity decreased, the paste was too hard and easy to clump, and it was not easy to apply. Taking all factors into consideration, it was finally determined that the appropriate dosage range of NP-700 was 0.8g-1.4g.

[0140] (2) Screening of the dosage of aluminum glycolate

[0141] according to Fig.13 The screening results show that when the dosage of aluminum glycolate is in the range of 0.2g-0.3g, the comprehensive score is higher. Fig.13 The results show that its dosage has a more obvious effect on the initial adhesion of the matrix, but has no significant effect on the lasting adhesion and sensory evaluation. When the dosage of glycyrrhizic aluminum exceeds 0.2g, the initial adhesion of the matrix shows a significant downward trend, resulting in a significant decrease in the comprehensive score of the matrix. With the increase in the dosage of glycyrrhizic aluminum, the cross-linking speed of the matrix becomes faster, the gel time is too short and the paste becomes too hard and difficult to apply. Taking all factors into consideration, it is finally determined that the appropriate dosage range of glycyrrhizic aluminum is 0.2g-0.3g.

[0142] (3) Screening of tartaric acid dosage

[0143] according to Fig.14 The screening results show that the matrix has a higher comprehensive score when the dosage of tartaric acid is in the range of 1.4g-1.8g. Fig.14 The effect of the dosage on the initial adhesion fluctuates greatly. After comprehensive consideration of various factors, the dosage range of tartaric acid was finally determined to be 1.4g-1.8g.

[0144] (4) Screening of dosage of sodium carboxymethyl cellulose

[0145] according to Fig.15 The screening results show that when the dosage of sodium carboxymethyl cellulose is in the range of 0.2g-0.6g, the comprehensive score of the matrix is ​​greater than 85 points. During the preparation of the matrix, it was found that when the dosage of sodium carboxymethyl cellulose exceeded 0.6g, the humidity of the paste became higher and higher, and the holding force and initial adhesion both decreased. Taking various factors into consideration, it was finally determined that the dosage of sodium carboxymethyl cellulose was 0.2g-0.6g.

[0146] (5) Screening of glycerol dosage

[0147] according to Fig.16 The screening results show that when the amount of glycerol is in the range of 8g-12g, the comprehensive score of the matrix is ​​higher. Fig.16It is shown that the amount of propylene glycol used has a great influence on the initial adhesion and lasting adhesion. When the amount of propylene glycol used is less than 6g, the auxiliary materials in the oil phase are too thick and difficult to disperse evenly, the paste clumps together and is not easy to apply and has low viscosity. When the amount used exceeds 8g, its initial adhesion is significantly improved. Considering various factors, the final range of propylene glycol used is determined to be 8g-12g.

[0148] The scoring standard of cataplasm was constructed by establishing a single-factor quality evaluation method, so as to select the target dosage with the best score. The quality evaluation of cataplasm includes the physicochemical properties and performance of cataplasm. The performance of cataplasm is evaluated according to the comprehensive score, which is the initial adhesion score + the lasting adhesion score + the sensory score. The evaluation process is as follows:

[0149] The physical and chemical properties of epidermal patches include color, shape and weight differences, and the performance of epidermal patches includes adhesion, fit, sustained adhesion and initial tack. Therefore, for quality evaluation, the initial adhesion and sustained adhesion of epidermal patches need to be measured to ensure that the patches have appropriate adhesion and sustained adhesion during use to meet the needs of clinical applications. At the same time, experiments are conducted to evaluate the performance of epidermal patches in combination with sensory evaluation and comprehensive scores of initial adhesion and sustained adhesion.

[0150] 1. Requirements for sensory index scoring

[0151]

[0152]

[0153] (II) Initial adhesion test

[0154] The initial adhesion is determined by the inclined ball rolling method, and the determination method is as follows: 1) Before the preparation test, the test sample should be placed at room temperature for more than 2 hours, and the surface of the inclined plate and the stainless steel ball should be wiped with a wiping material dipped in anhydrous ethanol, and then carefully wiped dry with a clean dust-free cloth. Repeat the cleaning for more than 3 times until the surface of the inclined plate and the stainless steel ball are clean after visual inspection; 2) Place the inclined ball rolling device in a horizontal position, and then adjust the inclination angle of the inclined plate to 30 degrees, take the poultice test sample and fix the backing of the poultice on the inclined plate respectively, where the upper end of the poultice should be located 100 cm below the horizontal line of the inclined plate, with the sticky side facing up and the poultice test sample should be flatly attached to the plate surface; 3) Place the steel ball specified under each variety in the groove at the top of the horizontal line and let it fall freely from the inclined surface; 4) Take 3 specimens and conduct 3 tests respectively.

[0155] Among the three pieces of the test product, two or more pieces should be able to stick to the steel ball. If one piece cannot stick, take a smaller steel ball for a test, and it should be able to stick. If one piece can stick to the steel ball and the other two pieces can stick to the smaller steel ball, take another three pieces for retest. If all three pieces can stick to the steel ball, it meets the requirements. The scoring method is: initial adhesion score = (test value / maximum value) × 30.

[0156] (III) Determination of holding strength

[0157] The determination method is as follows: 1) The test sample should be pasted on the test plate according to the prescribed method to ensure uniform pasting and no bubbles. Before pasting, the test plate and the loading plate should be cleaned to ensure the surface is clean; 2) The sample should be parallel to the longitudinal direction of the plate and pasted on the middle part of the adjacent test plate and loading plate; 3) Roll the sample with a pressure roller 3 times, then hang a steel plate on the tester and a 400g weight on the other, and record the time required for the poultice to slide until it falls off; The scoring method is: Adhesion score = (test value / maximum value) × 30.

[0158] Embodiment five:

[0159] According to the prepared colchicine diol plastid and the A phase and B phase solvents, a colchicine diol plastid cataplasm is prepared, and the preparation steps are as follows:

[0160] S1. Take an appropriate amount of colchicine diol plastid and add it to the prepared phase B solvent and stir to mix;

[0161] S2. Add the prepared phase A solvent to the mixed phase B solvent and mix again to obtain a mixed solution;

[0162] S3. Select several non-woven fabrics and evenly apply the obtained mixed solution on each non-woven fabric to prepare colchicine diol plasmid cataplasm;

[0163] according to Fig.17 The nonwoven fabric coated with colchicine diol plasmid cataplasm is shown in the figure. The prepared colchicine diol plasmid cataplasm is subjected to a skin irritation test. The skin irritation test includes:

[0164] (I) Single-dose skin irritation test

[0165] Two healthy adult New Zealand white rabbits of both sexes were selected, with a body weight of (2.03±0.22) kg. Four patches of hair were removed from both sides of the rabbits' backs with shaving scissors 24 hours before the experiment without damaging the skin. Each patch of hair removal had an area of ​​6 cm×8 cm, which was approximately 15% of the body surface area. The experimental animals were housed in separate cages, with one rabbit placed in each cage. They were fed pellet feed and had free access to drinking water.

[0166] During the experiment, the left hair removal area was treated: one hair removal area was selected for normal skin stimulation test, and the other hair removal area was selected for damaged skin stimulation test. A "#"-shaped scratch was made on the hair removal area skin with a sterilized needle to stimulate the epidermis without damaging the dermis. The degree of bleeding was observed.

[0167] During the experiment, the right side of the hair removal area was treated: one hair removal area was selected for normal skin stimulation test, and the other hair removal area was selected for damaged skin stimulation test. A "#"-shaped scratch was made on the hair removal area skin with a sterilized needle to stimulate the epidermis without damaging the dermis. The degree of bleeding was observed.

[0168] An equal amount of colchicine cataplasm was administered to the skin on the left side, and an equal amount of excipients was administered to the skin on the right side. After administration, the skin was covered with a film and two layers of sterile gauze, and fixed with adhesive tape. After 12 hours, the residual drug and excipients were washed off with gauze dipped in warm water. Then, the erythema and edema at the administration site on the 0th, 3rd, 5th, and 7th days were observed and recorded.

[0169] according to Fig.18 The single-dose skin irritation experiment showed that after discontinuation of the drug, the excipient group had no irritating effect on either intact or damaged skin of New Zealand white rabbits, and no erythema, edema or other abnormal phenomena occurred. In addition, the colchicine cataplasm group had no irritating effect on intact skin, and only very slight erythema and slight pigmentation appeared on damaged skin. However, as time went by, the erythema disappeared after the third day, indicating that a single administration of colchicine cataplasm was non-irritating to the skin.

[0170] (II) Repeated administration skin irritation test

[0171] Two healthy adult New Zealand white rabbits of both sexes were selected, with a body weight of (2.03±0.22) kg. Four patches of hair were removed from both sides of the rabbits' backs with shaving scissors 24 hours before the experiment without damaging the skin. Each patch of hair removal had an area of ​​6 cm×8 cm, which was approximately 15% of the body surface area. The experimental animals were housed in separate cages, with one rabbit placed in each cage. They were fed pellet feed and had free access to drinking water.

[0172] During the experiment, the left hair removal area was treated: one hair removal area was selected for normal skin stimulation test, and the other hair removal area was selected for damaged skin stimulation test. A "#"-shaped scratch was made on the hair removal area skin with a sterilized needle to stimulate the epidermis without damaging the dermis. The degree of bleeding was observed.

[0173] During the experiment, the right side of the hair removal area was treated: one hair removal area was selected for normal skin stimulation test, and the other hair removal area was selected for damaged skin stimulation test. A "#"-shaped scratch was made on the hair removal area skin with a sterilized needle to stimulate the epidermis without damaging the dermis. The degree of bleeding was observed.

[0174] An equal amount of colchicine cataplasm was administered to the skin on the left side, and an equal amount of vehicle was administered to the skin on the right side, and the drugs were administered once a day for one week. After stopping the drugs, the patients were observed for one week, and the occurrence of erythema, edema, pigmentation, hemorrhage spots and other abnormalities at the administration site on the 0th, 3rd, 5th and 7th days were recorded respectively;

[0175] according to Fig.19 The results of the multiple-dose skin irritation experiment show that the drug has no irritating effect on intact skin, and no edema or erythema occurs. When the excipient group is applied to damaged skin, slight erythema appears, which disappears over time. When the drug group is applied to damaged skin, erythema is barely visible, which basically disappears after the third day. This shows that the drug has no irritating effect on the skin when used for a long time, but long-term application to damaged skin should be avoided as much as possible.

[0176] Embodiment six:

[0177] In order to verify the efficacy of the prepared colchicine diol plasmid cataplasm, animal grouping and gout modeling experiments were conducted:

[0178] 1) 48 SD male rats, weighing 180-200 g, were selected and housed in non-toxic plastic boxes. The rats were fed with basic feed and tap water, and were allowed to eat freely and receive natural light. The litter in the plastic boxes was changed every two days.

[0179] 2) The rats were randomly divided into 6 groups, namely, a blank group, a model group, a colchicine oral tablet group, and three groups of low, medium, and high doses of colchicine diol plasmid cataplasm. All rats were depilated with depilatory cream on their legs, and the model was established 24 hours later;

[0180] 3) Except for the control group, rats in other groups were injected with 0.2 mL of sodium urate suspension (2.5 g / 100 mL) into the joint cavity between the middle of the ankle joint and the tibia and fibula using a sterile syringe, while rats in the control group were injected with an equal amount of normal saline at the same site;

[0181] 4) One hour after modeling, the rats in the colchicine oral tablet group were gavaged with colchicine tablet solution (0.5 mg / kg) once a day until the end of the experiment 48 hours after modeling. The rats in the colchicine diol plasmid cataplasm group were applied with a 4 cm thick patch on the ankle joint skin. 2 The best patch in vitro was fixed with medical tape and removed at the end of the experiment.

[0182] The circumference of the right hind ankle joint of the rats was observed and measured before modeling and 3h, 9h, 24h and 48h after modeling, and the joint swelling index of the rats in each group was calculated according to the following formula:

[0183]

[0184] In the above formula, L1 is the joint circumference at the measurement time point, and L0 is the initial joint circumference;

[0185] according to Fig. 20 From the data on the change in circumference value 0.5 mm below the right hind ankle joint of the rats in the model group, it can be seen that compared with the control group, after the injection of sodium urate solution, the left ankle joints of the rats in the model group were severely red and swollen, and the maximum swelling occurred after 12 hours. Subsequently, the redness and swelling of the joints of the rats in each group gradually improved. At the same time, it can be seen that the rats in the colchicine cataplasm group had a significant inhibitory effect on ankle swelling after drug intervention. Therefore, it is known that the therapeutic effect of colchicine cataplasm through transdermal absorption is similar to that of oral tablets.

[0186] The beneficial effects of the present invention are as follows: the colchicine diol plastid patch and the preparation method thereof are as follows: colchicine diol plastids and A phase solvent and B phase solvent are prepared respectively first, and then colchicine diol plastids are prepared according to the colchicine diol plastids and A phase solvent and B phase solvent, so that colchicine is prepared into a transdermal absorption preparation, and the skin permeability is further improved. After local skin administration, the drug can directly penetrate into the tissue from the skin surface without passing through the gastrointestinal tract, and reach the affected area after absorption, thereby achieving the effect of rapid therapeutic effect without causing systemic adverse reactions. By conducting multiple rat test processes, it is effectively verified that the cataplasm has good safety after contacting the skin. At the same time, combined with experimental data, it is known that the prepared colchicine diol plasmid patch has a certain therapeutic effect on gout, and is superior to traditional oral tablets in terms of treatment methods and effects. By combining the prepared colchicine diol plasmid patch with non-woven fabric, it is more convenient for patients to use it in the form of a patch, avoiding the situation where the ointment is easily scratched when applied and dried, and providing an isolated environment when the colchicine diol plasmid patch comes into contact with the skin, ensuring the purity of the drug during action and avoiding external pollution from affecting the efficacy. At the same time, a high waterproof material layer structure can be added to the non-attached surface of the non-woven fabric to further improve the environmental applicability of the colchicine diol plasmid patch.

[0187] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A colchicine diol plasmid cataplasm, characterized in that: It includes colchicine diol plasmid ingredients composed of soybean lecithin, cholesterol, colchicine, Tween-80, ethanol and propylene glycol, which are weighed and measured according to the prescription amount, and also includes a papule base ingredient composed of NP-700, glycerol, sodium carboxymethyl cellulose, diatomaceous earth, DL-tartaric acid, pure water and glycolylated aluminum, which are weighed in sufficient amount.

2. A colchicine diol plasmid according to claim 1, characterized in that: Colchicine diol plastids are prepared according to the colchicine diol plastid ingredients, and the preparation steps are as follows: L1. Place the soybean lecithin, cholesterol, colchicine and Tween-80 weighed according to the prescription amount into a 40 mL brown vial; L2. Then, add the ethanol and propylene glycol solutions according to the prescribed amount into a brown vial and sonicate for 20 minutes to fully dissolve them to obtain an alcohol phase solution; L3. After adding the magnet to the alcohol phase solution, place it in a constant temperature magnetic stirrer, set the stirring temperature to 20°C-50°C and the stirring speed to 500-800r / min for uniform stirring; L4. Add ceramide III powder to the alcohol phase solution at a concentration of 1 mg / mL and stir slowly to fully dissolve it; L5. Measure the PBS buffer solution according to the prescription, use a syringe to draw it as the aqueous phase test solution, and slowly drip the PBS buffer solution into the alcohol phase solution while continuing to hydrate at a constant temperature for 20 minutes; L6. After hydration, the mixture was filtered using a 0.22 μm filter membrane to prepare a colchicine diol plastid suspension having a blue opalescent luster and a light yellow color; The matrix is ​​prepared according to the ingredients of the cataplasm, and the preparation steps are as follows: T1. Take 8.12 g of glycerol in a beaker, and then evenly mix 1.27 g of NP-700 in glycerol to obtain a glycerol miscible solvent; T2. 0.6 g of sodium carboxymethyl cellulose and 0.2 g of diatomaceous earth were added to the glycerol miscible solvent and placed in a magnetic stirrer and stirred uniformly at a speed of 200 r / min to prepare a phase A solvent; T3. Take another beaker and weigh 1.56gDL-tartaric acid and 10g pure water and mix them evenly, then add 0.31g of aluminum glycolate and place the mixed solution in a magnetic stirrer; T4. Set the stirring speed to 200 r / min, the stirring temperature to 40° C. and the stirring time to 20 min to obtain the phase B solvent after uniform mixing.

3. A colchicine diol plasmid according to claim 2, characterized in that: The quality evaluation is carried out according to the method of controlling the variables of the bile-egg ratio, soybean lecithin dosage, dosage, diol ratio and alcohol phase dosage in the diol plastid ingredients, and the single factor screening of the dosage and process parameters of each formula is carried out according to the quality evaluation method. The evaluation content includes three determination methods of particle size, Zeta potential and encapsulation efficiency. Comprehensive scoring is carried out according to the evaluation content, and the scoring method is as follows: Comprehensive score = particle size score × 25% + Zeta potential score × 25% + encapsulation efficiency score × 50%; The evaluation samples were prepared according to the quality evaluation method, and the evaluation sample preparation method was as follows: 1) Select a 25 mL volumetric flask and quantitatively measure 1 mL of the prepared colchicine diol plasmid into the volumetric flask; 2) Add purified water to the colchicine diol plastids and dilute to the 25 mL mark. Shake well and then sonicate for 20 min to fully disperse the aggregated diol plastids to obtain a diluted solution sample.

4. A colchicine diol plasmid according to claim 3, characterized in that: The particle size determination and evaluation steps are as follows: a1. Select several ultrasonically cleaned particle size test dishes, take an appropriate amount of diluent sample and transfer the diluent to each particle size test dish to form several first determination sample liquids; a2. Measure three times in each first determination sample solution, take the average value of the obtained particle size data as the final result, and perform particle size scoring. The scoring method is as follows: The zeta potential determination and evaluation steps are as follows: b1. Select several ultrasonically cleaned potential test dishes, take an appropriate amount of diluent in the diluent sample and transfer the diluent to each potential test dish to form several copies of the second measurement sample solution; b2. Measure three times in each second determination sample solution, take the average value of the obtained Zeta potential data as the final result, and perform potential scoring, the scoring method is as follows:

5. A colchicine diol plasmid according to claim 3, characterized in that: The encapsulation efficiency determination and evaluation steps are as follows: c1. Select several 5mL volumetric flasks, measure 0.1mL of diol plasmid into each volumetric flask, and add methanol to mix until the 5mL mark; c2. Each mixed sample was vortexed for 10 minutes and then filtered through a 0.22 μm microporous membrane. At this time, the total colchicine concentration C of each mixed sample was measured by high performance liquid chromatography 总 , each sample was measured three times; c3. Select several dialysis bags with a molecular weight cutoff of 500, accurately measure 3 mL of colchicine diol plastids and transfer them to each dialysis bag, seal them with dialysis clips and place them in 150 mL of PBS solution; c4. Take out 5 mL of solution from each dialysis bag every 2 hours and add the same volume of solution at the same temperature until the concentration of the solution after dialysis remains unchanged; c5. Determine the concentration of colchicine in each dialyzed solution sample using a high performance liquid chromatograph 游 , each sample was measured three times; Among them C 游 represents the amount of free drug, i.e., the drug not encapsulated by the diol plastids; C 总 is the total amount of drug in the colchicine diol plastid suspension; The encapsulation efficiency is expressed as The encapsulation rate scoring method is as follows:

6. A method for preparing colchicine diol plasmid cataplasm, characterized in that: The invention comprises a colchicine diol plastosome cataplasm as described in claims 1 to 5, and further comprises the following preparation steps: S1. Take an appropriate amount of colchicine diol plastid and add it to the prepared phase B solvent and stir to mix; S2. Add the prepared phase A solvent to the mixed phase B solvent and mix again to obtain a mixed solution; S3. Select several non-woven fabrics and evenly apply the obtained mixed solution on each non-woven fabric to prepare colchicine diol plasmid patch.

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