A method for preparing an anti-inflammatory composition containing antimicrobial peptides by secondary homogenization

Abstract

The invention relates to the technical field of biopharmaceutical preparation, and is a method for preparing an anti-inflammatory composition containing an antimicrobial peptide through secondary homogenization. The invention creatively adds tachyplesin III antimicrobial peptide, which has excellent dual properties of cold resistance and high temperature resistance, meets the needs of high and low temperature secondary homogenization, and has high antimicrobial peptide activity after homogenization. The introduction of a modified gelatin solution can have a positive effect on the thermal stability of the product. The modified gelatin solution has hydrophilic groups and hydrophobic groups, and can form a stable phase with an aqueous phase, an oil phase and an emulsifier, and the phase will not be separated by the high and low temperature secondary homogenization. The high and low temperature secondary homogenization solves the problem of insufficient dispersibility after adding the antimicrobial peptide, artemisia annua extract and medicinal phytomycete extract. By arranging a homogenization auxiliary mechanism in the inner cavity of a homogenizing device, the oil and water floating on the upper layer are better mixed therein, which can better meet the actual homogenization work requirements and has a positive effect on improving product quality.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine preparation, and in particular to a method for preparing an anti-inflammatory composition containing antimicrobial peptides through secondary homogenization. Background Art

[0002] Antimicrobial peptides (AMPs) are a class of small molecule polypeptides composed of 5-100 amino acid residues with antimicrobial activity, which are encoded by specific genes of organisms and induced by external factors. They mainly come from plants, insects, amphibians, mammals, marine organisms, bacteria, viruses, etc. They are important defense systems in natural organisms. The antimicrobial activity of antimicrobial peptides is characterized by high selectivity, rapid bactericidal action, broad spectrum of action and difficulty in forming resistance, which makes them ideal candidates for the research and development of new antibiotics.

[0003] Currently, five types of horseshoe crab antimicrobial peptides have been discovered, namely tachyplesin I, tachyplesin II, tachyplesin III, Polyphemusin I and Polyphemusin II. Among them, tachyplesin I is the most widely studied and used. When testing the biological activity of genetically engineered antimicrobial peptides in a simulated human gastric and intestinal environment, it was found that tachyplesin I is stable below 100°C and pH less than 10.3. The cation concentration and solution polarity have a certain influence on the antimicrobial activity of tachyplesin I; when the tachyplesin I concentration is greater than 80 mg / L and the action time is greater than 30 minutes, it exhibits certain hemolytic activity; tachyplesin I shows high stability in simulated gastric fluid, gastric mucosal homogenate and plasma systems, the chromatographic peak shape is basically unchanged, and there is almost no degradation effect.

[0004] Therefore, tachyplesin I has strong high temperature tolerance, is stable under acidic conditions, and has a certain ability to withstand degradation by proteases in the body. However, tachyplesin I is not stable enough under low temperature conditions and is easily affected by external factors such as temperature, pH, and mechanical forces, resulting in poor activity.

[0005] However, there are relatively few studies on tachyplesinⅡ, tachyplesinⅢ, PolyphemusinI and PolyphemusinⅡ.

[0006] The anti-inflammatory cream containing antimicrobial peptides in the prior art has the following problems: 1. The absorption rate is low, resulting in waste of raw materials; 2. Since the cream is formed by an aqueous phase, an oil phase and an emulsifier, the cream is easily unstable after long-term storage and easily loses its effectiveness due to oil-water separation; the main drug ingredients are unevenly distributed in the cream, making the efficacy unstable.

[0007] The existing patent document with announcement number CN213643959U discloses a homogenizing device for the production of polypeptide drugs. The solution and the existing technology both use stirring and mixing to homogenize the drugs. However, it is difficult for the oil and water floating on the upper layer of the material to be fully mixed with the material at the bottom during mixing, making it difficult to achieve homogenization requirements. The existing mixing components lack a suitable auxiliary structure to optimize the mixing process.

[0008] To this end, the present invention provides an anti-inflammatory composition containing antimicrobial peptides and a preparation method for solving the above problems. Summary of the Invention

[0009] The object of the present invention is to provide an anti-inflammatory composition containing antimicrobial peptides and a preparation method thereof, so as to solve the problems raised in the above background technology.

[0010] To achieve the above objectives, the present invention provides the following technical solution: an anti-inflammatory composition containing an antimicrobial peptide, comprising, in parts by weight: 0.001-0.06 parts by weight of an antimicrobial peptide, 0.01-0.5 parts by weight of an Artemisia annua extract, 0.01-0.5 parts by weight of a medicinal phytomycete extract, 15-17 parts by weight of a water-based excipient, 15-18 parts by weight of an oil-based excipient, 30-60 parts by weight of a modified gelatin solution, 0.6-1 parts by weight of carbomer, and 136-140 parts by weight of water, wherein the antimicrobial peptide is tachyplesin III.

[0011] The water-based auxiliary material comprises disodium edetate, ethyl hydroxybenzoate, sorbitol and triethanolamine in a weight ratio of (0.01-0.02): (0.01-0.02): (10-15): (1.0-2.0) in that order;

[0012] The oil-based auxiliary material includes glyceryl stearate, polyethylene glycol stearate, peregal, octadecyl alcohol and hexadecanol, wherein the weight ratio is (6-8): (1-3): (1.5-3): (1.5-3): (1.5-3).

[0013] A method for preparing an anti-inflammatory composition containing an antimicrobial peptide comprises the following steps: first, mixing water, disodium edetate, and ethyl hydroxybenzoate; adding the modified gelatin solution and carbomer under stirring; stirring overnight; filtering; and then adding sorbitol and triethanolamine to the filtrate under stirring at a rate of 20-30 r / min; continuing stirring for 5 minutes; and heating to 65-75° C. to obtain an aqueous phase;

[0014] Step 2: Mix the oil-based auxiliary materials at 70-80°C and keep warm for 25-45 minutes to obtain an oil phase;

[0015] Step 3: mixing the antimicrobial peptide, the Artemisia annua extract and the medicinal phytomycetes extract in a mixer to obtain the main drug;

[0016] Step 4: After filtering the oil phase, vacuum-inhale it into an emulsification tank, mix it with the water phase at 28-35 r / min, homogenize it once at 70-80°C at 2300-2800 r / min through a homogenizer for 4-7 minutes, add the main drug and homogenize it a second time in a cold water bath at 5-10°C and 2300-2800 r / min for 2-4 minutes.

[0017] Preferably, the preparation method of modified gelatin includes: adding a surfactant sodium alkyl sulfate and deionized water, stirring and heating to 50°C, and after complete dissolution, adjusting the reaction pH to 8.0-9.0, adding gelatin, continuing to stir until the gelatin is completely dissolved, adding monoepoxy-terminated polysiloxane, reacting for 24 hours until the primary amine content no longer changes, stopping stirring and heating, and obtaining a modified gelatin solution.

[0018] Preferably, the preparation method of the modified gelatin comprises, in parts by mass: 0.1-0.5 parts of sodium alkyl sulfate, 90-105 parts of deionized water, 3-6g of gelatin, and the amount of monoepoxy-terminated polysiloxane used is 0.8 times the molar amount of primary amino groups in the gelatin molecule.

[0019] Preferably, the homogenizing device includes a homogenizing device body, a rotation control motor, a connecting shaft and a homogenizing auxiliary mechanism; a rotation control motor is provided on the homogenizing device body, one end of the rotation control motor is connected to the connecting shaft, the connecting shaft extends into the inner cavity of the homogenizing device body, and the connecting shaft is connected to the homogenizing auxiliary mechanism.

[0020] The control pump of described outer combustion gas heating unit is connected with the control pump of described outer combustion gas heating unit, and the control pump is provided with a pin on the bottom of described outer combustion gas heating unit; and the control pump is provided with a pin on the bottom of described outer combustion gas heating unit.

[0021] Preferably, the homogenizing stirring rod is hollow and is connected to the movable sleeve.

[0022] Preferably, the connecting vertical rod is hollow and connected to the homogenizing stirring rod, and the homogenizing stirring rod is perpendicular to the connecting vertical rod.

[0023] Preferably, the auxiliary component includes a cut-off ring body and a drainage auxiliary slope; the cut-off ring body is fixedly arranged on the inner side wall of the movable sleeve at equal distances, and the upper end surface of the cut-off ring body is arranged as the drainage auxiliary slope.

[0024] Preferably, the drainage auxiliary slope is arranged in a ring shape, and the drainage auxiliary slope corresponds to the position of the nozzle of the homogenizing stirring rod; the inner diameter of the intercepting ring body gradually decreases from top to bottom.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] On one hand, the present invention creatively contains tachyplesin III antimicrobial peptide, which has excellent dual properties of cold resistance and high temperature resistance, can meet the needs of high and low temperature secondary homogenization, and the antimicrobial peptide activity after homogenization is high;

[0027] On the other hand, the invention creatively introduces a modified gelatin solution, which can have a positive effect on the thermal stability of the product. The modified gelatin solution has hydrophilic and hydrophobic groups, which can form a stable phase with the water phase, oil phase and emulsifier, and the phase will not be subjected to secondary homogenization at high and low temperatures, resulting in stratification.

[0028] Furthermore, the present invention creatively adopts high and low temperature secondary homogenization to solve the problem of insufficient dispersibility after adding antimicrobial peptides, Artemisia annua extract and medicinal pore fungus extract, and utilizes a homogenizing device to further solve the problem of forming a stable phase between the water phase, the oil phase and the emulsifier, and the phase will not be stratified due to high and low temperature secondary homogenization; and by providing a corresponding homogenizing device in the preparation method and providing a homogenizing auxiliary mechanism in the inner cavity of the homogenizing device, the oil and water floating on the upper layer can be better mixed therein, which can better meet the actual homogenization work requirements and has a positive effect on improving product quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a tiling state diagram of a product according to an embodiment of the present invention;

[0030] Figure 2 This is a diagram showing the product of Comparative Example 1 of the present invention in a flattened state;

[0031] Figure 3 This is a diagram showing the product of Comparative Example 2 of the present invention in a flattened state;

[0032] Figure 4 This is a diagram showing the product of Comparative Example 3 of the present invention in a flattened state;

[0033] Figure 5 A top view of the structural connection of the homogenizing device of the present invention;

[0034] Figure 6 For the present invention Figure 5 A magnified schematic diagram of the structural connection at center A;

[0035] Figure 7 This is a schematic diagram of the front side of the homogenizing device structure connection of the present invention;

[0036] Figure 8 This is a schematic diagram of the connection between the rotating shaft, the homogenization auxiliary mechanism, and the auxiliary component structure of the present invention;

[0037] Figure 9 For the present invention Figure 8 A magnified schematic diagram of the structural connection at point B in the middle;

[0038] Figure 10 This is a schematic diagram of the connection between the homogenization auxiliary mechanism and the auxiliary components of the present invention;

[0039] Figure 11 For the present invention Figure 10 A magnified schematic diagram of the structural connection at point C in the middle;

[0040] Figure 12 For the present invention Figure 10 Enlarged schematic diagram of the structural connection at point D in the middle.

[0041] In the figure: homogenizing device body 1, rotation control motor 2, connecting shaft 3, fixed cylinder 401, delivery pipe 402, control pump body 403, rotating connector 404, movable sleeve 405, connecting bar 406, immersion pipe network 407, homogenizing stirring rod 408, first auxiliary hole 409, connecting vertical rod 410, second auxiliary hole 411, intercepting ring 501, drainage auxiliary slope 502. DETAILED DESCRIPTION

[0042] The technical solutions in the embodiments of the present invention are described clearly and completely below. The embodiments of the present invention and all other embodiments obtained by persons of ordinary skill in the art without creative work are within the scope of protection of the present invention.

[0043] In order to better understand the present invention, a specific embodiment is used for illustration. It is worth emphasizing that the effect of this embodiment is not substantially different from the various embodiments within the scope of protection of the present invention, including the respective reagents and the content ratio of the reagents. All of them can achieve the effects described in the present invention and solve the above-mentioned problems. Other combinations are not described here.

[0044] Example: An anti-inflammatory composition containing an antimicrobial peptide, comprising, by weight: 0.001-0.06 parts of an antimicrobial peptide, 0.01-0.5 parts of an Artemisia annua extract, 0.01-0.5 parts of a medicinal phytomycete extract, 15-17 parts of a water-based excipient, 15-18 parts of an oil-based excipient, 30-60 parts of a modified gelatin solution, 0.6-1 parts of carbomer, and 136-140 parts of water, wherein the antimicrobial peptide is tachyplesin III.

[0045] The water-based excipients are composed of disodium edetate, ethyl hydroxybenzoate, sorbitol and triethanolamine in the weight ratio of (0.01-0.02): (0.01-0.02): (10-15): (1.0-2.0) in sequence;

[0046] The oil-based auxiliary materials include glyceryl stearate, polyethylene glycol stearate, peregal, octadecyl alcohol and cetyl alcohol, wherein the weight ratio is (6-8): (1-3): (1.5-3): (1.5-3): (1.5-3).

[0047] A method for preparing an anti-inflammatory composition containing an antimicrobial peptide comprises the following steps: first, mixing water, disodium edetate, and ethyl hydroxybenzoate; adding a modified gelatin solution and carbomer while stirring; and filtering the mixture overnight. Then, adding sorbitol and triethanolamine to the filtrate while stirring at a rate of 20-30 r / min; continuing stirring for 5 minutes; and then heating the mixture to 65-75° C. to obtain an aqueous phase.

[0048] Step 2: Mix the oil-based auxiliary materials at 70-80°C and keep warm for 25-45 minutes to obtain the oil phase;

[0049] Step 3: mixing the antimicrobial peptide, the Artemisia annua extract and the medicinal phytomycetes extract in a mixer to obtain the main drug;

[0050] Step 4: Filter the oil phase and decompress it into the emulsification tank, mix it with the water phase at 28-35r / min, homogenize it once at 70-80℃ at 2300-2800r / min for 4-7min through a homogenizer, add the main drug and homogenize it a second time in a cold water bath at 5-10℃ and 2300-2800r / min for 2-4min.

[0051] Furthermore, the preparation method of modified gelatin includes: adding a surfactant sodium alkyl sulfate and deionized water, stirring and heating to 50°C, and after complete dissolution, adjusting the reaction pH to 8.0-9.0, adding gelatin, continuing to stir until the gelatin is completely dissolved, adding monoepoxy-terminated polysiloxane, reacting for 24 hours until the primary amine content no longer changes, stopping stirring and heating, and obtaining a modified gelatin solution.

[0052] Furthermore, the preparation method of the modified gelatin includes, in parts by mass: 0.1-0.5 parts of sodium alkyl sulfate, 90-105 parts of deionized water, 3-6 g of gelatin, and the amount of monoepoxy-terminated polysiloxane used is 0.8 times the molar amount of primary amino groups in the gelatin molecule;

[0053] Furthermore, the preparation method of Artemisia annua extract includes weighing a crushed sample, soaking the plant dry powder with 5 times the amount of ethanol, and standing at room temperature for 12 hours; ultrasonic extraction for 2 hours; vacuum filtration, continuous extraction 3 times, combining the filtrate, concentrating it with a rotary evaporator at 45°C, drying it, and storing it at 4°C for future use.

[0054] Furthermore, the preparation method of the medicinal pore fungus extract includes: crushing the medicinal pore fungus fruiting body and passing it through a 40-80 mesh sieve to obtain the medicinal pore fungus fruiting body powder, weighing the medicinal pore fungus fruiting body powder, adding 5 times the amount of 1,3-butanediol aqueous solution to the medicinal pore fungus fruiting body powder, stirring and extracting at a temperature of 60-70°C for 2-3h, filtering and collecting the filtrate, concentrating it with a rotary evaporator at 45°C, drying it, and storing it at 4°C for standby use.

[0055] Furthermore, a homogenizing device is used, and the homogenizing device is preferred. The homogenizing device includes a homogenizing device body 1, a rotation control motor 2, a connecting shaft 3 and a homogenizing auxiliary mechanism; a rotation control motor 2 is provided on the homogenizing device body 1, and one end of the rotation control motor 2 is connected to the connecting shaft 3, and the connecting shaft 3 extends into the inner cavity of the homogenizing device body 1, and the connecting shaft 3 is connected to the homogenizing auxiliary mechanism.

[0056] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0057] In this embodiment,

[0058] Step 1: First, water, disodium EDTA, and ethyl hydroxybenzoate were mixed, and modified gelatin solution and carbomer were added under stirring. The mixture was allowed to stand overnight, filtered, and sorbitol and triethanolamine were added to the filtrate under stirring at a rate of 20-30 r / min. The mixture was stirred for 5 minutes, and then heated to 65-75°C to obtain an aqueous phase.

[0059] Step 2: Mix the oil-based auxiliary materials at 70-80°C and keep warm for 25-45 minutes to obtain the oil phase;

[0060] Step 3: mixing the antimicrobial peptide, the Artemisia annua extract and the medicinal phytomycetes extract in a mixer to obtain the main drug;

[0061] Step 4: Filter the oil phase and decompress it into the emulsification tank, mix it with the water phase at 28-35r / min, homogenize it once at 70-80℃ at 2300-2800r / min for 4-7min, add the main drug and homogenize it for the second time, in a cold water bath, control it at 5-10℃ and 2300-2800r / min for 2-4min.

[0062] The examples and comparative examples were tested at the same level according to the following method:

[0063] (1) The product stability test standard is: the prepared cream is milky yellow, and under microscopic observation, the cream droplet size is less than 9 μm, uniformly dispersed in the cream matrix, and centrifuged at 3000 rpm for 30 minutes without stratification or demulsification;

[0064] (2) Stability test: Pour the same amount of the drug composition to be tested evenly into a culture dish and observe it at rest;

[0065] (2) Hot and cold cycle stability test

[0066] Test conditions: The samples were sealed in an oxygen-tight container and placed in the refrigerator's fresh-keeping compartment for 48 hours. After 48 hours, they were taken out and placed in a 40°C oven for 48 hours, with three cycles. After the test, the samples were subjected to a hot and cold cycle test using the same parameters as above, and the particle size was observed under a microscope.

[0067] Take out and place in a constant temperature box at 30℃ for long-term storage, and observe the particle size under a microscope;

[0068] In order to more intuitively demonstrate the stability, thermal stability and antioxidant process advantages of the present invention, the present invention and the same process using an equivalent replacement method are compared.

[0069] Comparative Example 1:

[0070] The preparation method is the same as that of the example, except that: in the preparation process of this comparative example, the antimicrobial peptide is replaced by tachyplesin III with tachyplesin I;

[0071] Comparative Example 2:

[0072] The preparation method is the same as that of the embodiment, except that: in the preparation process of this comparative example, no modified gelatin solution is added;

[0073] Comparative Example 3:

[0074] The preparation method is the same as that of the embodiment, except that: in the preparation process of this comparative example, gelatin solution of the same content and concentration purchased from the market is added;

[0075] Comparative Example 4:

[0076] The preparation method is the same as that of the embodiment, except that: in the preparation process of this comparative example, low-temperature one-step homogenization is adopted, and a gelatin solution of the same content and concentration purchased from the market is added;

[0077] The results are as follows:

[0078] Table 1: Experimental comparison of stability of modified gelatin and antimicrobial peptides in pharmaceutical compositions

[0079]

[0080]

[0081] According to the data analysis in Table 1:

[0082] (1) The results of the products tested in Example 1 and Comparative Example 1 show that the antimicrobial peptide is tachyplesin III antimicrobial peptide compared to other types of tachyplesin I; the anti-inflammatory composition containing tachyplesin III antimicrobial peptide has better dual properties of cold resistance and high temperature resistance, can meet the needs of high and low temperature secondary homogenization, and the antimicrobial peptide activity after homogenization is high and there is no agglomeration phenomenon, such as Figure 1 and Figure 2 ; The formation of agglomeration may be caused by the antimicrobial peptide tachyplesin Ⅰ being not resistant to high temperature.

[0083] (2) The results of the test products of Examples and Comparative Examples 1-4 show that the anti-inflammatory composition containing antimicrobial peptides prepared in the Examples of the present invention has stable properties and no crystal aggregation or agglomeration.

[0084] Among them, the products tested in Examples and Comparative Examples 1-3 were relatively stable in the short-term stability of 48 hours, and no stratification problem occurred. This may be due to the use of high and low temperature secondary homogenization to solve the problem of insufficient dispersibility after adding antimicrobial peptides, Artemisia annua extract and medicinal porphyromonas extract. High and low temperature secondary homogenization has better short-term stability than low temperature primary homogenization.

[0085] Comparative Example 4, which used low-temperature homogenization and no modified gelatin solution, also produced some stratification in the short-term stability of 48h, and did not meet the standard requirements of the product stability test.

[0086] This may be because this product is prepared by high and low temperature secondary homogenization of drug extracts, aqueous phase, oil phase and emulsifier, which meets the problem of insufficient dispersion of antimicrobial peptides, Artemisia annua extract and medicinal pore fungus extract in the main drugs. High and low temperature secondary homogenization has better short-term stability than low temperature primary homogenization; it is stable after long-term storage and is not easily separated by oil and water.

[0087] (3) After long-term stability, the anti-inflammatory compositions of Comparative Examples 2 and 3 showed obvious demulsification, oil-water separation, obvious centrifugal stratification, and crystal aggregation. Figure 3 and Figure 4 After the hot and cold cycle test, the hot and cold stability of the embodiment is stable, and there is no crystal aggregation phenomenon. Figure 1 .

[0088] Among them, the comparison between Example 3 and Comparative Example 3 shows that although the same amount of gelatin solution was added to Comparative Example 3, after the hot and cold cycle test, the cream showed obvious demulsification and oil-water separation, obvious centrifugal stratification, and crystal aggregation. The state of crystal aggregation is different from the agglomeration phenomenon of antimicrobial peptide denaturation at high temperature, and has a certain granular feel. The applicant speculates that this may be caused by the precipitation of complex active substances in the drug extract;

[0089] Therefore, the modified gelatin solution can produce a positive effect on the thermal stability of the product. The applicant infers that the modified gelatin solution has hydrophilic groups and hydrophobic groups, which can form a stable phase with the antimicrobial peptide, Artemisia annua extract, aqueous phase, oil phase and emulsifier, and this phase will not be affected by high temperature and cause stratification; while the gelatin solution has not undergone modification and therefore does not have the ability to form a stable phase with the drug extract, aqueous phase, oil phase and emulsifier, resulting in obvious demulsification and oil-water separation after the hot and cold cycle test, obvious centrifugal stratification, and especially the occurrence of crystal aggregation.

[0090] In summary:

[0091] (1) The present invention creatively adds the antimicrobial peptide tachyplesin III, which has excellent dual properties of cold resistance and high temperature resistance, can meet the needs of high and low temperature secondary homogenization, and the antimicrobial peptide activity after homogenization is high.

[0092] (2) The present invention creatively introduces high and low temperature secondary homogenization to solve the problem of insufficient dispersibility after adding antimicrobial peptides, Artemisia annua extract and medicinal porphyromonas extract to the main drug, and utilizes a homogenizing device to further solve the problem of forming a stable phase of water phase, oil phase and emulsifier, and this phase will not be separated by high and low temperature secondary homogenization, and has better stability in the short term;

[0093] (3) The invention creatively introduces modified gelatin solution, which can have a positive effect on the thermal stability of the product. The modified gelatin solution has hydrophilic groups and hydrophobic groups, and can form a stable phase containing antimicrobial peptides, Artemisia annua extract and medicinal pore fungus extract with the water phase, oil phase and emulsifier. In addition, the invention cooperates with high and low temperature secondary homogenization to solve the problem of insufficient dispersion of antimicrobial peptides, Artemisia annua extract and medicinal pore fungus extract in the main drug and maintain long-term stability. The use of a homogenizing device can further solve the problem of forming a stable phase with the water phase, oil phase and emulsifier, and the phase will not be subjected to high and low temperature secondary homogenization and cause stratification.

[0094] As a further study of the present invention, as embodiment 2 of the present invention: Based on embodiment 1, please refer to Figure 5-Figure 12, the homogenization auxiliary mechanism includes a fixed cylinder 401, a delivery pipe 402, a control pump body 403, a rotating connector 404, a movable sleeve 405, a connecting strip 406, an immersion pipe network 407, a homogenizing stirring rod 408, a first auxiliary hole 409, a connecting vertical rod 410, a second auxiliary hole 411 and an auxiliary component; one end of the fixed cylinder 401 is fixedly arranged on the top of the inner cavity of the homogenizing device body 1, and the other end of the fixed cylinder 401 is provided with a rotating connector 404, one side of the fixed cylinder 401 is connected to the delivery pipe 402, the delivery pipe 402 is connected to the control pump body 403, and the control pump body 403 is provided on the homogenizing device body 1, the other end of the rotating connector 404 is connected to one end of the movable sleeve 405, and the inner side wall of the bottom end of the movable sleeve 405 is equidistantly fixed with connecting strips 406, and the connecting strips 406 The other end is fixedly connected to the side wall of the connecting shaft 3, an immersion pipe network 407 is provided on the movable sleeve 405, a homogenizing stirring rod 408 is fixedly arranged at equal distances on the side wall of the movable sleeve 405, and a first auxiliary hole 409 is equidistantly provided on the homogenizing stirring rod 408, the other end of the homogenizing stirring rod 408 is arranged on the connecting vertical rod 410, and a second auxiliary hole 411 is equidistantly provided on the connecting vertical rod 410, and an auxiliary component is provided on the inner side wall of the movable sleeve 405; the homogenizing stirring rod 408 is hollow, and the homogenizing stirring rod 408 is connected to the movable sleeve 405; the connecting vertical rod 410 is hollow, and is hollow and connected to the homogenizing stirring rod 408, and the homogenizing stirring rod 408 is perpendicular to the connecting vertical rod 410; the auxiliary component includes a cut-off ring 501 and a drainage auxiliary slope 502.

[0095] In this embodiment, when the material is homogenized, the oil and water floating on the top layer are guided and mixed, and enter through the immersion pipe network 407 on the movable sleeve 405, and then drained downward through the movable sleeve 405, so as to enter the interior of the material faster and better. In this process, the intercepting ring body 501 is fixedly arranged on the inner wall of the movable sleeve 405 at equal distances, and the upper end face of the intercepting ring body 501 is set as a drainage auxiliary slope 502; the drainage auxiliary slope 502 is set in an annular shape, and the drainage auxiliary slope 502 corresponds to the pipe opening setting position of the homogenizing stirring rod 408; the inner diameter of the intercepting ring body 501 gradually decreases from top to bottom. Small; the oil and water flowing onto the intercepting ring 501 will enter the corresponding homogenizing stirring rod 408 and the connecting vertical rod 410 from the drainage auxiliary slope 502, and then enter the material through the first auxiliary hole 409 and the second auxiliary hole 411, which can achieve better diffusion and homogeneous mixing; a delivery pipe 402 is set on the delivery pipe 402, and the delivery pipe 402 here can convey the corresponding clean gas auxiliary material according to actual work needs, thereby improving its homogeneous mixing work efficiency; and when the material causes blockage to the first auxiliary hole 409 and the second auxiliary hole 411, it has a certain auxiliary effect on eliminating the blockage of the material.

Claims

1. A method for preparing an anti-inflammatory composition containing an antimicrobial peptide by secondary homogenization, characterized in that: The homogenizing device is used for secondary homogenization at high and low temperatures. The homogenizing device comprises a homogenizing device body (1), a rotation control motor (2), a connecting shaft (3), and a homogenizing auxiliary mechanism. The homogenizing device body (1) is provided with a rotation control motor (2), one end of the rotation control motor (2) is connected to the connecting shaft (3), the connecting shaft (3) extends into the inner cavity of the homogenizing device body (1), and the connecting shaft (3) is connected to the homogenizing auxiliary mechanism. The homogenizing auxiliary mechanism includes a fixed cylinder (401), a delivery pipe (402), a control pump body (403), a rotating connector (404), a movable sleeve (405), a connecting strip (406), an immersion pipe network (407), a homogenizing stirring rod (408), a first auxiliary hole (409), a connecting vertical rod (410), a second auxiliary hole (411) and an auxiliary component; The auxiliary component includes a flow-cutting ring body (501) and a drainage auxiliary slope (502); the flow-cutting ring body (501) is fixedly arranged on the inner side wall of the movable sleeve (405) at equal distances, and the upper end surface of the flow-cutting ring body (501) is arranged as the drainage auxiliary slope (502); The inner side wall of the bottom end of the movable sleeve (405) is fixedly provided with a connecting strip (406) at equal intervals, and the other end of the connecting strip (406) is fixedly connected to the side wall of the connecting shaft (3). The movable sleeve (405) is provided with an immersion pipe network (407). The homogenizing stirring rod (408) is fixedly provided at equal intervals on the side wall of the movable sleeve (405). The homogenizing stirring rod (408) is provided with a first auxiliary hole (409) at equal intervals. The other end of the homogenizing stirring rod (408) is provided on the connecting vertical rod (410). The connecting vertical rod (410) is provided with a second auxiliary hole (411) at equal intervals. The inner side wall of the movable sleeve (405) is provided with an auxiliary component. The homogenizing stirring rod (408) is hollow and is connected to the movable sleeve (405). The connecting vertical rod (410) is hollowed out, the drainage auxiliary slope (502) is annular, and the drainage auxiliary slope (502) corresponds to the position of the nozzle of the homogenizing stirring rod (408).

2. The method for preparing an anti-inflammatory composition containing an antimicrobial peptide by secondary homogenization according to claim 1, characterized in that: One end of the fixed cylinder (401) is fixedly arranged at the top of the inner cavity of the homogenizing device body (1), and the other end of the fixed cylinder (401) is provided with a rotating connector (404). One side of the fixed cylinder (401) is connected to a delivery pipe (402), and the delivery pipe (402) is connected to a control pump body (403), and the control pump body (403) is provided on the homogenizing device body (1). The other end of the rotating connector (404) is connected to one end of a movable sleeve (405).

3. The method for preparing an anti-inflammatory composition containing an antimicrobial peptide by secondary homogenization according to claim 1, characterized in that: The connecting vertical rod (410) is connected to the homogenizing stirring rod (408), and the homogenizing stirring rod (408) is perpendicular to the connecting vertical rod (410).

4. The method for preparing an anti-inflammatory composition containing an antimicrobial peptide by secondary homogenization according to claim 1, characterized in that: The inner diameter of the intercepting ring (501) gradually decreases from top to bottom.

5. The method for preparing an anti-inflammatory composition containing an antimicrobial peptide by secondary homogenization according to any one of claims 1 to 4, characterized in that: The specific steps of preparing the antimicrobial peptide-containing anti-inflammatory composition by secondary homogenization using a homogenizer are as follows: Step 1: first, water, disodium edetate, and ethyl hydroxybenzoate are mixed, and modified gelatin solution and carbomer are added under stirring, overnight, filtered, and sorbitol and triethanolamine are added to the filtrate under stirring speed of 20-30 r / min, and stirring is continued for 5 minutes, and then heated to 65-75°C to obtain an aqueous phase; Step 2: oil-based excipients are mixed at 70-80°C and kept warm for 25-45 minutes to obtain an oil phase; Step 3: Step 3: Mix the antimicrobial peptide, Artemisia annua extract and medicinal pore fungus extract in a mixer to obtain the main drug; Step 4: Filter the oil phase and decompress it into an emulsification tank, mix it with the water phase at 28-35 r / min, homogenize it once at 70-80°C and 2300-2800 r / min through a homogenizer for 4-7 minutes, add the main drug and homogenize it for a second time, and control it in a cold water bath at 5-10°C and 2300-2800 r / min for 2-4 minutes.

6. The method for preparing an anti-inflammatory composition containing antimicrobial peptides by secondary homogenization according to claim 5, characterized in that: The preparation method of modified gelatin includes: adding a surfactant sodium alkyl sulfate and deionized water, stirring and heating to 50°C, adjusting the reaction pH to 8.0-9.0 after complete dissolution, adding gelatin, continuing to stir until the gelatin is completely dissolved, adding monoepoxy-terminated polysiloxane, reacting for 24 hours until the primary amine content no longer changes, stopping stirring and heating, and obtaining a modified gelatin solution.

7. The method for preparing an anti-inflammatory composition containing an antimicrobial peptide by secondary homogenization according to claim 6, characterized in that: The invention comprises, by weight, 0.001-0.06 parts of antimicrobial peptide, 0.01-0.5 parts of Artemisia annua extract, 0.01-0.5 parts of medicinal pore fungus extract, 15-17 parts of water-based excipients, 15-18 parts of oil-based excipients, 30-60 parts of modified gelatin solution, 0.6-1 parts of carbomer, and 136-140 parts of water, wherein the antimicrobial peptide is tachyplesin III, water The base excipients are composed of disodium edetate, ethyl hydroxybenzoate, sorbitol and triethanolamine in a weight ratio of (0.01-0.02): (0.01-0.02): (10-15): (1.0-2.0) respectively; the oil-based excipients include glyceryl stearate, polyethylene glycol stearate, peregal, octadecanol and cetyl alcohol, in a weight ratio of (6-8): (1-3): (1.5-3): (1.5-3): (1.5-3).

8. The method for preparing an anti-inflammatory composition containing antimicrobial peptides by secondary homogenization according to claim 6, characterized in that: The preparation method of the modified gelatin comprises the following parts by mass: 0.1-0.5 parts of sodium alkyl sulfate, 90-105 parts of deionized water, 3-6g of gelatin, and the amount of monoepoxy-terminated polysiloxane used is 0.8 times the molar amount of primary amino groups in the gelatin molecule.

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