Compound sulfonamide metformin sodium nanometer mixed micelle solution and preparation method thereof

Abstract

The present application belongs to the field of veterinary drug preparation, and particularly relates to a compound sulfonamide metronidazole sodium nanometer mixed micelle solution and a preparation method thereof. The preparation method is as follows: omeprazole and trimethoprim are jointly coated in mixed micelles formed by hydroxypropyl methyl cellulose derivatives and polyethylene glycol 1000 vitamin E succinate to obtain a micelle solution; then the micelle solution is mixed with an aqueous solution containing sulfonamide metronidazole sodium and an antioxidant, and is uniformly mixed, and the pH is adjusted to 9.5-11.5, and after sterilization, the compound sulfonamide metronidazole sodium nanometer mixed micelle solution is obtained. The prepared compound sulfonamide metronidazole sodium nanometer mixed micelle solution has good aqueous solution stability, can be used with water at will, is suitable for use in large-scale breeding farms, does not contain organic solvents, reduces the irritation in clinical use, is used for preventing and treating chicken E. coli and Salmonella infection, can have a long-term effect, and has a good treatment effect.

Description

Technical Field

[0001] This invention belongs to the field of veterinary drug preparations, specifically relating to a compound sulfamethoxypyrimidine sodium nanomixed micelle solution and its preparation method. Background Technology

[0002] Sulfamonomethoxine (SMM) is a novel sulfonamide drug with the strongest in vitro and in vivo antibacterial activity. It interferes with bacterial metabolic processes, blocking the enzyme that synthesizes dihydrofolate, thereby inhibiting bacterial growth and reproduction. SMM has a strong inhibitory effect on most Gram-positive and Gram-negative bacteria, and has advantages such as a broad antibacterial spectrum, rapid absorption, high stability, resistance to deterioration, wide availability, and low price. It is mainly used for various diseases caused by susceptible bacteria, and has good efficacy in treating infections and coccidiosis in poultry such as chickens caused by susceptible bacteria. To enhance the efficacy of sulfonamide drugs, they are often used in combination with antibacterial synergists, which can achieve a dual blocking effect on bacterial folic acid metabolism.

[0003] Omeprazole (OMP), as a novel antibacterial potentiator, has a longer half-life, higher bioavailability, better synergistic effect with antibiotics, and lower toxicity, which can greatly reduce the dosage of antibiotics and alleviate the side effects of drug abuse. However, because omeprazole itself is almost insoluble in water and has poor absorption, uneven mixing or food can affect its efficacy. If it were made into a soluble form and administered through drinking water, it would be easier to administer and help improve drug absorption and utilization. Omeprazole can form salts with organic or inorganic acids to improve water solubility, but when this form of soluble salt is used in combination with soluble sulfonamides (usually alkaline sodium salts), the antagonistic effect can cause mutual interference, resulting in a decrease in the solubility of both, and in severe cases, precipitation, which affects the efficacy of the medication. Trimethoprim (TMP), an antibacterial synergist, also suffers from poor water solubility and low bioavailability. Most commercially available water-soluble trimethoprim is its lactate salt, which is acidic. When used in combination with alkaline sulfonamide salts, it produces a large amount of precipitation, which seriously affects the efficacy.

[0004] Currently, most commercially available compound sulfamethoxypyrimidine sodium solutions use organic solvents to dissolve trimethoprim. When the product has a high concentration in water, it will precipitate out, making it unsuitable for administration via dosing devices in intensive farms. Therefore, improving the solubility of poorly soluble antibacterial synergists will help advance the research and development of combined use of antibacterial synergists and antibacterial drugs. Summary of the Invention

[0005] The purpose of this invention is to address the shortcomings of existing technologies by providing a compound sulfamethoxypyrimidine sodium nano-mixed micelle solution and its preparation method. This preparation method is simple to operate, operates under mild conditions, and reduces the amount of organic solvents used. The resulting compound sulfamethoxypyrimidine sodium nano-mixed micelle solution exhibits good aqueous solution stability, can be easily diluted with water, and is suitable for use in large-scale poultry farms. Its in vitro release behavior is relatively slow and prolonged, allowing for sustained efficacy. Furthermore, it does not contain organic solvents, reducing irritation in clinical use. When used to prevent and treat Escherichia coli and Salmonella infections in chickens, it provides a long-lasting effect and good therapeutic efficacy.

[0006] The technical solution adopted by the present invention to achieve the above objectives is as follows:

[0007] A method for preparing a compound sulfamethoxypyrimidine sodium nanomixed micelle solution, comprising the following steps (parts by weight):

[0008] S1. Add 0.2-0.3 parts of hydroxypropyl methylcellulose derivative and compound 1 to 15-25 parts of water, mix thoroughly to obtain a mixed solution; the chemical structural formula of compound 1 is:

[0009]

[0010] S2. Add 0.5-1 parts of trimethoprim, 0.5-1 parts of omeprine and 2-3 parts of polyethylene glycol 1000 vitamin E succinate to an organic solvent, sonicate for 5-10 min, remove the organic solvent by vacuum distillation, then slowly add the mixed solution, sonicate for 20-30 min, and filter using a 0.22 μm microporous membrane to obtain a micelle solution.

[0011] S3. Dissolve 5-10 parts of sodium sulfamethoxypyrimidine and 0.2-0.4 parts of antioxidant in 15-25 parts of water, mix well, then mix well with micelle solution, adjust pH to 9.5-11.5 using alkaline adjuster, and sterilize to obtain the final product.

[0012] Further, the preparation method of the hydroxypropyl methylcellulose derivative in step S1 is as follows: vitamin E succinate, hydroxypropyl methylcellulose, and p-toluenesulfonic acid are added sequentially to a reactor, heated to 100-110℃, and reacted for 4-6 hours to obtain the hydroxypropyl methylcellulose derivative; the mass ratio of vitamin E succinate, hydroxypropyl methylcellulose, and p-toluenesulfonic acid is 1:3-5:0.5-0.6.

[0013] Further, the amount of compound 1 used in step S1 is 0.2-0.5% of the mass of the mixed solution; the preparation method of compound 1 is as follows:

[0014] (1) Imidazole, 4-chloro-1-butanol, and triethylamine were added to N,N-dimethylformamide, heated to 80-90℃, and reacted for 8-10 h. After the reaction was completed, the mixture was naturally cooled to room temperature, filtered, and saturated sodium sulfate aqueous solution and dichloromethane were added to the filtrate for extraction. The organic phase was collected, dried, and concentrated under reduced pressure to obtain compound 2. The molar ratio of imidazole, 4-chloro-1-butanol, and triethylamine was 1:1.1-1.2:1.5-2.0. The reaction process was as follows:

[0015]

[0016] (2) Maleic anhydride and n-heptanol are added to a reaction vessel, heated to 60°C to melt the maleic anhydride, and then the temperature is increased to 85-95°C. The reaction is carried out for 5-6 hours to obtain compound 3. The molar ratio of maleic anhydride to n-hexanol is 1:1-1.1. The reaction process is as follows:

[0017]

[0018] (3) Compounds 2 and 3 are added to dichloromethane, heated to 30-40℃, and reacted for 13-15 hours to obtain compound 1; the molar ratio of compounds 2 and 3 is 1:1-1.1; the reaction process is as follows:

[0019]

[0020] Furthermore, the organic solvent mentioned in step S2 is one or a mixture of several of methanol, ethanol, acetone, chloroform, and acetonitrile.

[0021] Furthermore, the antioxidant mentioned in step S3 is one or a mixture of several of sodium thiosulfate, sodium bisulfite, and vitamin C; the basic regulator is sodium hydroxide.

[0022] This invention provides a compound sulfamethoxypyrimidine sodium nanomixed micelle solution, which is prepared by the method described above for preparing the compound sulfamethoxypyrimidine sodium nanomixed micelle solution.

[0023] The present invention has the following beneficial effects:

[0024] This invention uses imidazole as a starting material to synthesize an imidazole-based surfactant compound 1. Experimental results show that compound 1 has good ability to reduce oil-water interfacial tension and good wetting and emulsifying properties. This invention uses compound 1 in the preparation of nano-mixed micelles, which enables more uniform dispersion of drugs and coating materials in water, helping to improve drug loading and encapsulation efficiency. This invention also uses hydroxypropyl methylcellulose and vitamin E succinate as raw materials to synthesize a hydroxypropyl methylcellulose derivative in a one-step solvent-free process. The molecular structure of the obtained hydroxypropyl methylcellulose derivative contains vitamin E succinate groups, which have a partially identical structure with polyethylene glycol 1000 vitamin E succinate. This improves the compatibility between the hydroxypropyl methylcellulose derivative and polyethylene glycol 1000 vitamin E succinate, facilitating stable drug coating under the wetting and emulsifying dispersion effects of surfactant compound 1, thereby obtaining a nano-mixed micelle solution with a smooth, regular surface and uniform size.

[0025] This invention encapsulates omeprine and trimethoprim, which have poor water solubility, in a mixed micelle formed by hydroxypropyl methylcellulose derivative and polyethylene glycol 1000 vitamin E succinate. This improves the solubility of omeprine and trimethoprim, increases their bioavailability, and helps enhance their synergistic effect on sulfamethoxypyrimidine sodium. Moreover, the nano-mixed micelle solution prepared by this invention has a certain sustained-release effect, with a relatively slow and prolonged in vitro release behavior, enabling it to exert its efficacy for a longer period of time.

[0026] The preparation method of compound sulfamethoxypyrimidine sodium nano-mixed micelle solution provided by this invention is simple to operate, mild under mild conditions, and reduces the amount of organic solvent used. The prepared compound sulfamethoxypyrimidine sodium nano-mixed micelle solution has good aqueous solution stability, can be diluted with water at will, and is suitable for use in large-scale farms. It does not contain organic solvents, reducing irritation in clinical use. It can be used to prevent and treat Escherichia coli and Salmonella infections in chickens, and has a long-lasting effect and good therapeutic effect. Attached Figure Description

[0027] Figure 1 The graph shows the relationship between the mass fraction of compound 1 and interfacial tension.

[0028] Figure 2 Transmission electron microscopy image of a nano-mixed micelle solution of compound sulfamethoxypyrimidine sodium;

[0029] Figure 3 The in vitro release curves of the compound sulfamethoxypyrimidine sodium nanomixed micelle solution are shown. Detailed Implementation

[0030] The technical solutions in the embodiments of this application will be clearly and completely described below with reference to the embodiments of this application. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0031] Sulfamethoxypyrimidine sodium CAS No. 1037-50-9; Trimethoprim CAS No. 738-70-5; Omeprine CAS No. 6981-18-6; Polyethylene glycol 1000 Vitamin E succinate CAS No. 9002-96-4; Sodium hydroxide CAS No. 1310-73-2; Sodium thiosulfate CAS No. 7772-98-7; Sodium bisulfite CAS No. 7631-90-5; Vitamin C CAS No. 50-81-7; Hydroxypropyl methylcellulose CAS No. 9004-65-3 (viscosity 5 mPa·s); Vitamin E succinate CAS No. 4345-03-3; All chemical reagents are commercially available.

[0032] Example 1

[0033] A method for preparing a compound sulfamethoxypyrimidine sodium nanomixed micelle solution, comprising the following steps (parts by weight):

[0034] S1. Add 0.3 parts of hydroxypropyl methylcellulose derivative and compound 1 to 25 parts of water, mix well to obtain a mixed solution; wherein the amount of compound 1 is 0.5% of the mass of the mixed solution;

[0035] S2. Add 1 part trimethoprim, 1 part omeprine and 3 parts polyethylene glycol 1000 vitamin E succinate to an organic solvent, sonicate for 10 min, remove the organic solvent by vacuum distillation, then slowly add the mixed solution, sonicate for 30 min, and filter using a 0.22 μm microporous membrane to obtain a micelle solution; wherein the organic solvent is ethanol.

[0036] S3. Dissolve 10 parts of sodium sulfamethoxypyrimidine and 0.4 parts of antioxidant in 25 parts of water, mix well, then mix well with micelle solution, adjust the pH to 9.5 using an alkaline adjuster, and sterilize to obtain the final product; wherein the antioxidant is sodium thiosulfate and the alkaline adjuster is sodium hydroxide.

[0037] The preparation method of hydroxypropyl methylcellulose derivative is as follows: vitamin E succinate, hydroxypropyl methylcellulose, and p-toluenesulfonic acid are added sequentially to a reactor, heated to 110℃, and reacted for 5 hours. After the reaction is completed, the mixture is naturally cooled to room temperature, ethyl acetate is added, and the mixture is extracted three times with saturated brine. The organic phase is collected and concentrated under reduced pressure to obtain the hydroxypropyl methylcellulose derivative. The mass ratio of vitamin E succinate, hydroxypropyl methylcellulose, and p-toluenesulfonic acid is 1:5:0.6, and the amount of ethyl acetate used is 30 times the mass of vitamin E succinate.

[0038] The preparation method of compound 1 is as follows:

[0039] (1) 15.0 g imidazole, 26.3 g 4-chloro-1-butanol, and 33.4 g triethylamine were added to 1000 mL N,N-dimethylformamide, heated to 90 °C, and reacted for 8 h. After the reaction was completed, the mixture was allowed to cool naturally to room temperature, filtered, and 1000 mL saturated sodium sulfate aqueous solution and 2000 mL dichloromethane were added to the filtrate for extraction. The organic phase was collected, dried, and concentrated under reduced pressure to obtain 28.5 g of compound 2. The molar ratio of imidazole, 4-chloro-1-butanol, and triethylamine was 1:1.1:1.5. The reaction process was as follows:

[0040]

[0041] Compound 2: ESI (m / z): 141.1 [M+H] + , 1 H-NMR (600MHz, DMSO-d6, δppm): 7.92 (s, 1H), 7.17 (d, 1H), 6.78 (d, 1H), 4.14 (s, 1H), 3.86-3.91 (m, 4H), 1.95 (m, 2H), 1.52 (m, 2H).

[0042] (2) Add 15.0 g of maleic anhydride and 17.8 g of n-heptanol to the reaction vessel, heat to 60 °C to melt the maleic anhydride, and continue heating to 90 °C. React for 6 h to obtain 30.2 g of compound 3; the molar ratio of maleic anhydride to n-hexanol is 1:1; the reaction process is as follows:

[0043]

[0044] Compound 3: ESI (m / z): 215.1 [M+H] + , 1 H-NMR (600MHz, DMSO-d6, δppm): 14.35 (s, 1H), 6.52 (d, 1H), 6.32 (d, 1H), 3.92-3.95 (m, 2H), 1.61-1.63 (m, 2H), 1.28-1.37 (m, 8H), 0.89 (t, 3H).

[0045] (3) 10.0 g of compound 2 and 15.3 g of compound 3 were added to dichloromethane, heated to 35 °C, and reacted for 14 h to obtain 22.8 g of compound 1; the molar ratio of compound 2 to compound 3 was 1:1; the reaction process was as follows:

[0046]

[0047] Compound 1: ESI (m / z): 372.2 [M+H] + , 1 H-NMR (600MHz, DMSO-d6, δppm): 8.91 (s, 1H), 7.91 (d, 1H), 7.76 (d, 1H), 4.38-4.40 (m, 2H), 4.15 (s, 1H), 3.49- 3.54(m, 4H), 3.40(m, 1H), 3.30-3.15(m, 2H), 1.48-1.53(m, 6H), 1.34(m, 2H), 1.26-1.28(m, 6H), 0.89(t, 3H).

[0048] The surface activity of the prepared compound 1 was tested as follows:

[0049] The oil-water interfacial tension test was conducted by preparing an aqueous solution of compound 1 with a mass fraction of 1×10⁻⁶. -6 2×10 -6 3×10 -6 4×10 -6 5×10 -6 6×10 -6 The interfacial tension between oil and water was measured, with physiological saline as the aqueous solvent and aviation kerosene as the oil phase solvent; the results are as follows. Figure 1 As shown, at a mass fraction of 1×10 -6 -6×10 -6 Compound 1 can reduce the oil-water interfacial tension to 0.5 × 10⁻⁶. -2 -2.6×10 -2 The mN / m indicates that compound 1 can achieve low oil-water interfacial tension over a wide concentration range, demonstrating its good ability to reduce oil-water interfacial tension and indicating good wetting properties.

[0050] The emulsifying ability test was conducted as follows: A 0.1% (w / w) aqueous solution of imidazole surfactant compound 1, sodium dodecylbenzenesulfonate, and Tween 80 was prepared. 20 mL of the prepared aqueous solution of compound 1, sodium dodecylbenzenesulfonate, and Tween 80, and 20 mL of paraffin oil or n-hexane were added to a 100 mL stoppered graduated cylinder. The cylinder was then tightly sealed with a glass stopper and vigorously shaken up and down approximately 30 times. After standing horizontally for 2 minutes, the mixture was shaken 30 times again and allowed to stand for 2 minutes. This shaking and standing process was repeated 5-6 times. It was observed that the aqueous and oil phases in the mixture gradually separated. The time required to separate 10 mL of water was recorded as a standard for measuring the emulsifying performance; a longer time indicates a stronger emulsifying ability of the surfactant. The results are shown in Table 1.

[0051] Table 1. Emulsifying performance test results of compound 1

[0052] Compound 1 462 434 Sodium dodecylbenzenesulfonate 305 336 Twain 80 371 354

[0053] As shown in Table 1, the water separation time of surfactant Compound 1 is longer than that of sodium dodecylbenzenesulfonate and Tween 80, indicating that the emulsifying performance of Compound 1 is significantly better than that of ordinary surfactants (sodium dodecylbenzenesulfonate and Tween 80). The results show that Compound 1 has good emulsifying performance.

[0054] Example 2

[0055] A method for preparing a compound sulfamethoxypyrimidine sodium nanomixed micelle solution, comprising the following steps (parts by weight):

[0056] S1. Add 0.2 parts of hydroxypropyl methylcellulose derivative and compound 1 to 15 parts of water, mix well to obtain a mixed solution; wherein the amount of compound 1 is 0.2% of the mass of the mixed solution;

[0057] S2. Add 0.5 parts of trimethoprim, 0.5 parts of omeprine and 2 parts of polyethylene glycol 1000 vitamin E succinate to an organic solvent, sonicate for 5 min, remove the organic solvent by vacuum distillation, then slowly add the mixed solution, sonicate for 20 min, and filter using a 0.22 μm microporous membrane to obtain a micelle solution; wherein the organic solvent is acetone.

[0058] S3. Dissolve 10 parts of sodium sulfamethoxypyrimidine and 0.2 parts of antioxidant in 15 parts of water, mix well, then mix well with micelle solution, adjust the pH to 10 using an alkaline adjuster, and sterilize to obtain the final product; wherein the antioxidant is sodium bisulfite and the alkaline adjuster is sodium hydroxide.

[0059] The preparation methods of the hydroxypropyl methylcellulose derivative and compound 1 are the same as those in Example 1.

[0060] Example 3

[0061] A method for preparing a compound sulfamethoxypyrimidine sodium nanomixed micelle solution, comprising the following steps (parts by weight):

[0062] S1. Add 0.25 parts of hydroxypropyl methylcellulose derivative and compound 1 to 20 parts of water, mix well to obtain a mixed solution; wherein the amount of compound 1 is 0.4% of the mass of the mixed solution;

[0063] S2. Add 0.8 parts of trimethoprim, 0.8 parts of omeprine and 2.5 parts of polyethylene glycol 1000 vitamin E succinate to an organic solvent, sonicate for 8 min, remove the organic solvent by vacuum distillation, then slowly add the mixed solution, sonicate for 25 min, and filter using a 0.22 μm microporous membrane to obtain a micelle solution; wherein the organic solvent is methanol.

[0064] S3. Dissolve 8 parts of sodium sulfamethoxypyrimidine and 0.3 parts of antioxidant in 20 parts of water, mix well, then mix well with micelle solution, adjust the pH to 11.5 using an alkaline adjuster, and sterilize to obtain the final product; wherein the antioxidant is vitamin C and the alkaline adjuster is sodium hydroxide.

[0065] The preparation methods of the hydroxypropyl methylcellulose derivative and compound 1 are the same as those in Example 1.

[0066] Example 4

[0067] A method for preparing a compound sulfamethoxypyrimidine sodium nanomixed micelle solution, comprising the following steps (parts by weight):

[0068] S1. Add 0.3 parts of hydroxypropyl methylcellulose derivative and compound 1 to 20 parts of water, mix well to obtain a mixed solution; wherein the amount of compound 1 is 0.3% of the mass of the mixed solution;

[0069] S2. Add 1 part trimethoprim, 0.5 parts omeprine and 2 parts polyethylene glycol 1000 vitamin E succinate to an organic solvent, sonicate for 8 min, remove the organic solvent by vacuum distillation, then slowly add the mixed solution, sonicate for 25 min, and filter using a 0.22 μm microporous membrane to obtain a micelle solution; wherein the organic solvent is acetonitrile.

[0070] S3. Dissolve 10 parts of sodium sulfamethoxypyrimidine and 0.2 parts of antioxidant in 20 parts of water, mix well, then mix well with micelle solution, adjust the pH to 11 using an alkaline adjuster, and sterilize to obtain the final product; wherein the antioxidant is vitamin C and the alkaline adjuster is sodium hydroxide.

[0071] The preparation methods of the hydroxypropyl methylcellulose derivative and compound 1 are the same as those in Example 1.

[0072] Comparative Example 1

[0073] The method for preparing the mixed solution differs from that in Example 1.

[0074] A method for preparing a compound sulfamethoxypyrimidine sodium nanomixed micelle solution, comprising the following steps (parts by weight):

[0075] S1. Add 0.3 parts of hydroxypropyl methylcellulose and compound 1 to 15-25 parts of water, mix well, and obtain a mixed solution;

[0076] S2, Same as step S2 in Example 1;

[0077] S3, Same as step S3 in Example 1;

[0078] The preparation method of compound 1 is the same as that in Example 1.

[0079] Comparative Example 2

[0080] The method for preparing the mixed solution differs from that in Example 1.

[0081] A method for preparing a compound sulfamethoxypyrimidine sodium nanomixed micelle solution, comprising the following steps (parts by weight):

[0082] S1. Add 0.3 parts of hydroxypropyl methylcellulose derivative to 25 parts of water, mix well to obtain a mixed solution;

[0083] S2, Same as step S2 in Example 1;

[0084] S3, Same as step S3 in Example 1;

[0085] The preparation method of the hydroxypropyl methylcellulose derivative is the same as that in Example 1.

[0086] Comparative Example 3

[0087] Compound sulfamethoxypyrimidine sodium solution was prepared using the raw material formulation and method described in Specific Embodiment 2 of Patent Document CN103006674B.

[0088] The preparation method is as follows:

[0089] The ingredients are: 10g sulfamethoxypyrimidine sodium, 2g trimethoprim, 50ml organic solvent, 0.2g antioxidant, and 0.2g metal complex; wherein the organic solvent is a solution obtained by mixing propylene glycol and benzyl alcohol in a 4:1 ratio, the antioxidant is sodium bisulfite, and the complex is disodium ethylenediaminetetraacetate; the specific steps are as follows:

[0090] (1) Place the organic solvent in a beaker and heat it in a water bath at 60-70°C. Pour the weighed sodium sulfamethoxypyrimidine and trimethoprim into the beaker and stir continuously until the mixture becomes clear.

[0091] (2) Take another beaker, add an appropriate amount of water, preheat it in a water bath at 30-70℃, add antioxidants and complexing agents, and stir constantly to make it clear;

[0092] (3) Slowly add the solution in (2) to the solution in (1) and stir continuously to mix them evenly to obtain a clear solution;

[0093] (4) After cooling, add the remaining water to the solution obtained in (3) and make up to 100 ml.

[0094] (5) Adjust the pH to 9.5-11.5 with sodium hydroxide solution.

[0095] The prepared compound sulfamethoxypyrimidine sodium solution was sterilized by steam sterilization at 120°C for 15 minutes.

[0096] Related experiments

[0097] 1. Detection of the apparent morphology, particle size, and zeta potential of nano-mixed micelle solutions

[0098] The nano-mixed micelle solutions prepared in Examples 1-4 and Comparative Examples 1-2 were subjected to morphology, particle size, and zeta potential analysis. The morphology analysis method was as follows: a drop of sample was placed on a copper grid and negatively stained with 2.0% phosphotungstic acid solution, then the morphology of the nanomicelles was observed using a transmission electron microscope. Particle size and zeta potential analysis: an appropriate amount of nano-micelle solution was taken, and the particle size and zeta potential were measured using a Malvern Nanozs 90 laser particle size analyzer. The results are shown in Table 2. Figure 2 This is a transmission electron microscope (TEM) image of the compound sulfamethoxypyrimidine sodium nanomixed micelle solution prepared in Example 1.

[0099] Table 2. Detection results of the apparent morphology, particle size, and zeta potential of the nano-mixed micelle solution.

[0100] Example 1 It is spherical in shape, with a smooth and regular surface and uniform size. 75±5 -14 Example 2 It is spherical in shape, with a smooth and regular surface and uniform size. 80±4 -15 Example 3 It is spherical in shape, with a smooth and regular surface and uniform size. 90±8 -16 Example 4 It is spherical in shape, with a smooth and regular surface and uniform size. 95±6 -16 Comparative Example 1 The particles are not round and are uneven in size. 120±15 -8 Comparative Example 2 The particles are not round and are uneven in size. 150±20 -10

[0101] As shown in Table 2, the nano-mixed micelle solutions prepared in Examples 1-4 have a consistent appearance, i.e., they are spherical with rounded and regular surfaces and uniform size; the particle size distribution is in the range of 70-110 nm, and the Zeta potential distribution is in the range of -16 to -14. Compared with Comparative Examples 1 and 2, the nano-mixed micelles prepared in Example 1 are made using hydroxypropyl methylcellulose derivatives. The molecular structure of the hydroxypropyl methylcellulose derivative contains vitamin E succinate groups, which have a partially identical structure with polyethylene glycol 1000 vitamin E succinate. This makes the compatibility between the hydroxypropyl methylcellulose derivative and polyethylene glycol 1000 vitamin E succinate better, which helps to form a stable coating of the drug under the wetting, emulsifying and dispersing effect of surfactant compound 1, thereby obtaining a nano-mixed micelle solution with rounded and regular surfaces and uniform size.

[0102] 2. Detection of drug loading and encapsulation efficiency of nano-mixed micelle solutions

[0103] Drug loading and encapsulation efficiency determination: The drug loading and encapsulation efficiency of the compound sulfamethoxypyrimidine sodium mixed nanomicelle solution were determined by ultra-high performance liquid chromatography (UPLC). A Diamonsil C18 column was used, with a detection wavelength of 280 nm. The mobile phase was a mixture of methanol and 0.02 mol / L potassium dihydrogen phosphate (30:70). The column temperature was 25℃, the flow rate was 1.0 mL / min, and the injection volume was 10 μL. The determination method is as follows:

[0104] Accurately weigh sulfamethoxazole sodium reference standard and add it to the mobile phase to prepare a sulfamethoxazole sodium reference standard solution containing 50 μg / mL per 1 mL; accurately weigh omeprine and trimethoprim reference standards respectively and add them to the mobile phase to prepare omeprine and trimethoprim reference standard solutions containing 50 μg / mL per 1 mL.

[0105] Dilute each reference solution 1:20 with the mobile phase, accurately inject 10 μL into the chromatograph, and record the chromatogram. Calculate the contents of sulfamethoxazole sodium, omeprine, and trimethoprim using the external standard method based on peak area, and convert the values ​​proportionally to the actual contents.

[0106] An appropriate amount of nanomicelle solution was weighed using an electronic balance, dissolved in methanol, and sonicated for 5 minutes to completely destroy the core-shell structure of the nanomicelles, releasing trimethoprim and omeprine from the nanomicelle solution. The actual content of trimethoprim and omeprine released from the solution was then measured using ultra-high performance liquid chromatography (UHPLC). The drug loading percentage (DL%) was calculated as: (weight of drug in micelles / weight of drug-loaded micelles) × 100%; the encapsulation efficiency (EE%) was calculated as: (weight of drug in micelles / amount of drug added) × 100%. The results are shown in Table 3.

[0107] Table 3. Results of drug loading and encapsulation efficiency of nano-mixed micelle solutions.

[0108] Example 1 16.13 97.04 Example 2 14.10 96.00 Example 3 13.08 95.03 Example 4 15.05 96.01 Comparative Example 1 10.34 79.10 Comparative Example 2 8.52 71.65

[0109] As shown in Table 3, the nano-mixed micelle solutions prepared in Examples 1-4 have high drug loading and encapsulation efficiency. Compared with Comparative Examples 1 and 2, the surfactant compound 1 used in Example 1, which has good wetting, emulsifying and dispersing effects, can make the components disperse evenly in water. This helps to encapsulate the drug with hydroxypropyl methylcellulose derivatives that have good compatibility with polyethylene glycol 1000 vitamin E succinate, thereby improving the drug loading and encapsulation efficiency.

[0110] 3. Solubility and stability tests of nano-mixed micelle solutions

[0111] To determine the solubility of the nano-mixed micelle solution in water, 1 mL of the product from Examples 1-4 and Comparative Examples 1-3 was mixed with 10 mL of water and thoroughly mixed (the concentration was 100 times the high-dose concentration used in clinical applications). The mixture was then inverted 10 times at 25℃ ± 2℃ and allowed to stand for 30 min before observing the water solubility and precipitation. The results are shown in Table 4.

[0112] Table 4. Solubility and stability test results of nano-mixed micelle solutions

[0113] Example 1 Colorless and clear Colorless and clear Colorless and clear Colorless and clear Colorless and clear Colorless and clear Example 2 Colorless and clear Colorless and clear Colorless and clear Colorless and clear Colorless and clear Colorless and clear Example 3 Colorless and clear Colorless and clear Colorless and clear Colorless and clear Colorless and clear Colorless and clear Example 4 Colorless and clear Colorless and clear Colorless and clear Colorless and clear Colorless and clear Colorless and clear Comparative Example 1 Colorless and clear Colorless and clear Colorless and clear Colorless and clear Crystal precipitation Crystal precipitation Comparative Example 2 Colorless and clear Colorless and clear Colorless and clear Crystal precipitation Crystal precipitation Crystal precipitation Comparative Example 3 Crystal precipitation Crystal precipitation Crystal precipitation Crystal precipitation Crystal precipitation Crystal precipitation

[0114] As shown in Table 4, the nano-mixed micelle solutions prepared in Examples 1-4 have good aqueous solution stability and remain colorless and clear after prolonged standing. This indicates that the present invention uses hydroxypropyl methylcellulose derivatives to encapsulate drugs under the action of surfactant compound 1, which can prepare nano-mixed micelle solutions with good stability.

[0115] 4. In vitro release experiment of nano-mixed micelle solution

[0116] In vitro release experiments of nano-mixed micelle solutions were conducted using dialysis. The total content of trimethoprim and omepramine in the release medium was measured using ultra-high performance liquid chromatography (UHPLC). 2 mL of the nano-mixed micelle solutions prepared in Examples 1-4, Comparative Examples 1-2, and Comparative Example 3 were added to 1 mL of water and placed in dialysis bags (MW = 8000-140000). After sealing, the bags were immersed in Erlenmeyer flasks containing 15 mL of release medium (PBS, pH = 7.4, containing 0.5% Tween 80) and placed in a 37°C constant temperature water bath shaker at 100 rpm. At preset time points, 1 mL samples were taken, and the same volume of fresh release medium was added simultaneously. The samples were filtered through a 0.22 μm microporous membrane, and the total content of trimethoprim and omepramine was determined. The cumulative release percentage was calculated, and a curve of the cumulative release percentage versus time was plotted, i.e., the in vitro drug release curve. Figure 3 As shown in the release curve, the conventional compound sulfamethoxypyrimidine sodium solution (Comparative Example 3) was basically completely released after 25 hours, while the drug-loaded nano-mixed micelle solution (Example 1, Comparative Examples 1-2) had a sustained-release effect. In particular, the nano-mixed micelle solution prepared by the present invention had a better sustained-release effect, with a cumulative release percentage of over 70% after 60 hours, and the release behavior was relatively slow and persistent.

[0117] 5. Animal experiments with nano-mixed micelle solutions

[0118] In a large-scale breeding center, 420 chickens infected with Escherichia coli were randomly selected as experimental subjects and divided into 7 groups of 60 chickens each. The products prepared in Examples 1-4 and Comparative Examples 1-2 were administered to the chickens in their drinking water at a concentration of 80 mg / L (based on the mass of pure sulfamethoxypyrimidine). The treatment was continued for 3 days, during which the chickens were fed normally. After 2 days of drug withdrawal, the treatment status of the chickens in each group was observed. The results are shown in Table 5.

[0119] Table 5. Treatment effects on sick chickens in each group

[0120] Example 1 100 Normal stool, good spirits, increased feed intake Example 2 100 Normal stool, good spirits, increased feed intake Example 3 100 Normal stool, good spirits, increased feed intake Example 4 100 Normal stool, good spirits, increased feed intake Comparative Example 1 86 There is mild diarrhea, but the feed intake is normal. Comparative Example 2 85 There is mild diarrhea, but the feed intake is normal. Comparative Example 3 74 There was mild diarrhea, poor appetite, and low food intake.

[0121] As shown in Table 5, the compound sulfamethoxypyrimidine sodium nano-mixed micelle solution prepared in Examples 1-4 has a significantly better therapeutic effect on chickens infected with Escherichia coli than that in Comparative Examples 1-3, with a cure rate of 100%. This indicates that the compound sulfamethoxypyrimidine sodium nano-mixed micelle solution prepared in this invention has a good therapeutic effect on common chicken diseases.

[0122] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Although embodiments of this application have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of this application, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a compound sulfamethoxypyrimidine sodium nanomixed micelle solution, characterized in that, By weight, the following steps are included: S1. Add 0.2-0.3 parts of hydroxypropyl methylcellulose derivative and compound 1 to 15-25 parts of water, mix thoroughly to obtain a mixed solution; the chemical structural formula of compound 1 is: ； S2. Add 0.5-1 parts of trimethoprim, 0.5-1 parts of omeprine and 2-3 parts of polyethylene glycol 1000 vitamin E succinate to an organic solvent, sonicate for 5-10 min, remove the organic solvent by vacuum distillation, then slowly add the mixed solution, sonicate for 20-30 min, and filter using a 0.22 μm microporous membrane to obtain a micelle solution. S3. Dissolve 5-10 parts of sodium sulfamethoxypyrimidine and 0.2-0.4 parts of antioxidant in 15-25 parts of water, mix well, then mix well with micelle solution, adjust pH to 9.5-11.5 with alkaline adjuster, and sterilize to obtain the final product. The preparation method of the hydroxypropyl methylcellulose derivative in step S1 is as follows: vitamin E succinate, hydroxypropyl methylcellulose, and p-toluenesulfonic acid are added sequentially to a reactor, heated to 100-110℃, and reacted for 4-6 hours to obtain the hydroxypropyl methylcellulose derivative; the mass ratio of vitamin E succinate, hydroxypropyl methylcellulose, and p-toluenesulfonic acid is 1:3-5:0.5-0.6; the amount of compound 1 used in step S1 is 0.2-0.5% of the mass of the mixed solution; The antioxidant mentioned in step S3 is one or a mixture of several of sodium thiosulfate, sodium bisulfite, and vitamin C; the basic regulator is sodium hydroxide.

2. The method for preparing a compound sulfamethoxypyrimidine sodium nanomixed micelle solution according to claim 1, characterized in that, The organic solvent mentioned in step S2 is one or a mixture of several of methanol, ethanol, acetone, chloroform, and acetonitrile.

3. A compound sulfamethoxypyrimidine sodium nanomixed micelle solution, characterized in that, It was prepared by the method described in any one of claims 1-2 for preparing the compound sulfamethoxypyrimidine sodium nanomixed micelle solution.

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