A polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft, solid dispersion of Coptis chinensis extract, preparation method and application

Abstract

The present invention relates to a polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft, a solid dispersion of Coptis chinensis extract, and a preparation method and application thereof. The structure of the polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft is shown in formulas (I) and (II). The present invention provides a preparation method of the polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft and its application as a drug carrier. The present invention also provides a solid dispersion of Coptis chinensis extract and a preparation method and application thereof. The present invention provides a novel drug carrier, which can mask the bitterness of the Coptis chinensis extract, and at the same time enable the Coptis chinensis extract to pass through the gastrointestinal tract for dissolution, solving the problem of gastric irritation caused by the dissolution of the oral Coptis chinensis extract in the stomach, and also solving the problem that the existing solid dispersion of Coptis chinensis extract is prepared at high temperature, which easily leads to the deterioration and degradation of the active ingredient berberine hydrochloride in the Coptis chinensis extract.

Description

Technical Field

[0001] The present invention relates to the technical field of veterinary drug preparations, and particularly relates to a polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft, a solid dispersion of Coptis extract, and a preparation method and application thereof. Background Art

[0002] Coptis Extract (CE) has various effects such as antibacterial, anti-inflammatory, antioxidant, hypoglycemic, and liver protection, and is commonly used for the prevention and treatment of piglet diarrhea. The active ingredient of CE is berberine hydrochloride (BBR·Hcl), and its absorption method is mainly intestinal absorption. When administered by intramuscular or intravenous injection, it will cause strong adverse reactions. Therefore, oral administration of CE is safer and more effective. However, CE has an extremely bitter taste and poor palatability, and its strong bitterness cannot even be masked by sweeteners. If directly mixed with feed, animal refusal to eat is very common, seriously affecting the drug efficacy. At the same time, CE has a certain gastric irritation after oral administration, causing an increase in gastric acid and damaging the gastric mucosa, thus causing gastric discomfort and even gastric ulcers.

[0003] Currently, the treatment technology of CE does not completely mask the bitterness and dissolves in the stomach. For example, a solid dispersion of Coptis extract, in which 10% of the CE solid dispersion dissolves in 90 minutes in oral solution and more than 80% dissolves in 2 minutes in gastric solution, does not completely mask the bitterness of CE and has strong gastric irritation. Another example is an oral preparation containing Coptis extract with bitterness masking, which treats CE with a cation exchange resin with a sulfonic group. Although it masks the bitterness of CE, it does not solve the problem of CE dissolving in the stomach.

[0004] In addition, to solve the problems of palatability, reduce gastric release, and reduce gastric irritation, some researchers have prepared CE into enteric-coated preparations with a coating layer. For example, an enteric-coated Coptis extract pellet / a berberine enteric-coated pellet, but these processes only physically coat the pellets made of CE. Although it can dissolve in the intestinal environment and solve the adverse reactions caused by CE absorption in the stomach, during the chewing process of animals, it is still possible to chew the pellets, thereby releasing the bitter taste of Coptis, affecting animal feeding. Moreover, the pellet diameter is relatively large and the mixing with feed is uneven, which will affect the drug efficacy.

[0005] A solid-intestine antidiarrheal enteric-coated traditional Chinese medicine preparation, which prepares a traditional Chinese medicine formula containing Coptis components into a solid-intestine antidiarrheal enteric-coated traditional Chinese medicine preparation. Although it has good solubility in the intestine and can increase the dissolution and release of the preparation in the intestine, avoiding the influence of the gastrointestinal tract, this enteric-coated preparation is in the form of capsules, granules, pellets, and dripping pills. These preparations have relatively large particles and are not easy to mix evenly with feed during feed mixing administration, affecting the efficacy.

[0006] Solid dispersions (SD) can mask the bitter taste and unpleasant odor of drugs and improve the compliance of animals in taking medicine. At present, however, the melting method is used to prepare CE into solid dispersions. For example, a solid dispersion of Coptis chinensis extract uses a hot melt extrusion process, which requires heating both CE and the carrier to the molten state. Although the melting temperature of the active ingredient BBR·Hcl in CE is about 200-206°C, BBR·Hcl is thermally unstable and will oxidize, deteriorate, and change color at 80°C and above. Therefore, preparing CE SD by the hot melt extrusion process is likely to cause the deterioration and degradation of the active ingredient BBR·Hcl, thereby affecting the clinical effect of the drug. Another example is the solid dispersion of Coptis chinensis detoxifying powder, which stirs and heats the CE-containing solution at 80-95°C, which will cause the thermal degradation of BBR·Hcl and affect the drug efficacy. At the same time, this process also does not solve the problem of gastric irritation caused by oral administration of CE. Summary of the Invention

[0007] The purpose of the present invention is to provide a polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft, a solid dispersion of Coptis chinensis extract, a preparation method and an application thereof, so as to provide a new drug carrier to effectively mask the bitter taste of CE, and at the same time enable CE to pass through the gastrointestinal tract and dissolve, so as to solve the problem of gastric irritation caused by the dissolution of oral CE in the stomach, and can also solve the problem that the existing solid dispersion of Coptis chinensis extract is prepared at high temperature, which is likely to cause the deterioration and degradation of the active ingredient BBR·Hcl of CE.

[0008] In order to achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0009] A polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft, the structure of which is a compound shown in formula (Ⅰ) and (Ⅱ) and its stereoisomers, tautomers or pharmaceutically acceptable salts thereof:

[0010] [[ID=1—14]] ,

[0011] ,

[0012] In formula (Ⅰ) and (Ⅱ), the value of n is a natural number greater than or equal to 1. The specific value of n is positively correlated with the weights of vitamin E polyethylene glycol 1000 succinate (TPGS) and EudragitL100 / S100.

[0013] The polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft of the present invention forms an amphiphilic structure by utilizing the hydrophilic PEG-1000 (polyethylene glycol 1000) chain of TPGS and the hydrophobic vitamin E, and thus self-assembles into micelles or jointly forms nanoparticles with Eudragit L100 / S100. When it is used as a novel drug carrier, it can effectively encapsulate the API CE, thereby masking the bitterness of CE, reducing the aggregation of CE in the gastrointestinal tract or blood, improving the delivery efficiency, and enabling CE to pass through the gastro-intestinal dissolution, solving the problem of gastric irritation caused by the dissolution of oral CE in the stomach.

[0014] Among them, CE used in the present invention is obtained by direct purchase, and the mass percentage content of berberine hydrochloride in CE is greater than or equal to 3.6%.

[0015] The present invention also provides a preparation method of the polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft, comprising the following steps:

[0016] Dissolve vitamin E polyethylene glycol 1000 succinate (TPGS) in dimethyl sulfoxide (DMSO), then add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), stir at room temperature for 6 h, then rotary evaporate to remove the unreacted EDC / NHS, and freeze-dry to obtain the activated vitamin E polyethylene glycol 1000 succinate;

[0017] Add Eudragit L100 and Eudragit S100 to a mixed solvent of absolute ethanol and acetone, heat to dissolve, then add the activated vitamin E polyethylene glycol 1000 succinate, heat and react under an inert atmosphere, and filter to obtain the polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft (i.e., Eudragit L100 / S100-TPGS graft).

[0018] Among them, the obtained polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft is a mixture of structural formulas (Ⅰ) and (Ⅱ).

[0019] According to the above technical means, by utilizing the esterification reaction between the carboxyl groups of Eudragit L100 / S100 and the hydroxyl groups of TPGS to form ester bonds, TPGS is grafted onto Eudragit L100 / S100, and an Eudragit L100 / S100-TPGS graft is innovatively prepared. The enteric-coated CE SD prepared with this graft can effectively avoid the significant solubilization effect of the surfactant TPGS on CE in a pH 1.2 hydrochloric acid solution (simulating gastric juice), and at the same time increase its cumulative dissolution rate in a pH 6.8 phosphate buffer solution (simulating intestinal juice), resulting in a low dissolution rate of CE in the stomach and rapid disintegration and dissolution in the intestine.

[0020] Preferably, the molar ratio of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to N-hydroxysuccinimide is 1.2:1.

[0021] Preferably, the mass ratio of Eudragit L100, Eudragit S100, and activated vitamin E polyethylene glycol 1000 succinate is 2:1:0.08 to 1:2:0.12.

[0022] Preferably, the volume ratio of absolute ethanol to acetone is 9:1.

[0023] Preferably, the temperature for heating and dissolving is 45-55 °C.

[0024] Preferably, the temperature for heating and reacting under an inert atmosphere is 48-52 °C, and the time is 45-50 h.

[0025] Preferably, the inert atmosphere is a nitrogen atmosphere.

[0026] The present invention also provides an application of the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft prepared by the above preparation method as a drug carrier.

[0027] Preferably, the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft is used as an enteric carrier.

[0028] The present invention also provides a solid dispersion of Coptis chinensis extract (CE SD), which includes Coptis chinensis extract and the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft prepared by the preparation method of the present invention.

[0029] The CE SD of the present invention is prepared from CE wrapped by Eudragit L100 / S100-TPGS. After the CE SD is released in the intestine, the surface-active effect of TPGS can reduce the surface tension of the mucosa, enhance the penetration of the drug through the intestinal epithelium, and thus improve the bioavailability of orally administered CE. At the same time, the antioxidant property (vitamin E) of TPGS is used to protect the drug CE from oxidative degradation, and the rigid skeleton of Eudragit L100 / S100 can also enhance the physical stability of CE, effectively prolonging the storage time. As an excipient approved by the FDA, TPGS has high biocompatibility, can neutralize the mucosal irritation that may be caused by orally administered Eudragit L100 / S100, and improve the safety of drug use.

[0030] The present invention also provides a preparation method of a solid dispersion of Coptis chinensis extract, comprising the following steps:

[0031] Add the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft copolymer into an ethanol aqueous solution, after heat treatment, add the Coptis chinensis extract in batches, then carry out heat-up treatment, low-temperature distillation, solidification, drying, pulverization, and sieving to obtain the CESD powder.

[0032] In the preparation method of the CE SD of the present invention, by first adding the graft copolymer into the ethanol aqueous solution for heat treatment, the graft copolymer is fully dissolved or dispersed to form a uniform solution or dispersion system. At this time, the temperature of the solution is relatively low and will not damage BBR·HCl in CE. Then the CE is added in batches, and then heat-up treatment is carried out. However, this heat-up treatment is carried out on the basis of the graft copolymer having formed a stable system, and the temperature is controlled within a suitable range to avoid damage to BBR·HCl caused by too high a temperature. It solves the problem that the existing solid dispersion of Coptis chinensis extract is prepared at high temperature, which easily leads to the deterioration and degradation of the active ingredient BBR·HCl of CE. And the preparation method has the advantages of simple process, convenient operation, and low cost. The required equipment is conventional equipment in veterinary drug factories, which is convenient for large-scale industrial production, has great economic and social benefits, and meets the requirements of green ecology and sustainable development.

[0033] The CE SD prepared by the present invention is in powder form, will not release bitterness due to animal chewing, has good palatability, stable properties, good homogeneity, has good fluidity, and is convenient for mixing with feed for administration.

[0034] Preferably, the temperature of the heat treatment is 48-52 °C.

[0035] Preferably, the temperature of the heat-up treatment is 58-62 °C.

[0036] Preferably, the temperature of the low-temperature distillation is less than 60 °C. Among them, the pressure of the low-temperature distillation is normal pressure.

[0037] Preferably, the mass ratio of the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft to the Coptis chinensis extract is 6-8:2-4.

[0038] Preferably, the volume ratio of absolute ethanol to water in the ethanol aqueous solution is 95:5.

[0039] Preferably, the curing is to place the residue after distillation under the condition of -25~-15°C for 20~28h for curing.

[0040] Preferably, the drying is to dry the cured block under vacuum conditions.

[0041] The present invention also provides an application of the CE SD in a drug for preventing and treating animal diarrhea.

[0042] Preferably, the administration method of the CE SD is oral administration.

[0043] The beneficial effects of the present invention:

[0044] For the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft of the present invention, by utilizing the hydrophilic PEG-1000 chain of TPGS and the hydrophobic vitamin E to form an amphiphilic structure, it self-assembles into micelles or jointly forms nanoparticles with Eudragit L100 / S100. When used as a carrier, it can effectively encapsulate the API CE, thereby masking the bitterness of CE, reducing the aggregation of CE in the gastrointestinal tract or blood, improving the delivery efficiency, and enabling CE to pass through the gastro-intestinal solution, solving the problem of gastric irritation caused by the dissolution of oral CE in the stomach.

[0045] For the preparation method of the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft of the present invention, by utilizing the carboxyl group of Eudragit L100 / S100 and the hydroxyl group of TPGS to undergo an esterification reaction to form an ester bond, TPGS is grafted onto Eudragit L100 / S100, and the Eudragit L100 / S100-TPGS graft is innovatively prepared. The enteric-coated CE SD prepared with this graft can effectively avoid the significant solubilization effect of the surfactant TPGS on CE in a pH 1.2 hydrochloric acid solution (simulating gastric juice), and at the same time increase its cumulative dissolution rate in a pH 6.8 phosphate buffer solution (simulating intestinal juice), resulting in a low dissolution rate of CE in the stomach and rapid disintegration and dissolution in the intestine.

[0046] The preparation method of CE SD of the present invention is to first add the grafted product into an ethanol aqueous solution for heat treatment to fully dissolve or disperse the grafted product, forming a uniform solution or dispersion system. At this time, the temperature of the solution is relatively low and will not damage BBR·HCl in CE. Then, CE is added in batches, and then the temperature is raised. However, this temperature raising treatment is carried out on the basis that the grafted product has formed a stable system, and the temperature is controlled within a suitable range to avoid damage to BBR·HCl caused by too high a temperature.

[0047] In addition to the raw material drug CE, the carrier of CE SD of the present invention is only a grafted product prepared from an enteric material and a surfactant. It has a high drug loading. On the basis of having no coating layer structure, it overcomes the disadvantages of poor palatability and large gastric irritation of CE at the same time. And CE SD is in powder form, will not release bitterness due to animal chewing, has good palatability, stable properties, good uniformity, has good fluidity, is convenient for mixing with feed for administration, and has the value of popularization and application in the technical field of veterinary drug preparations. Brief Description of the Drawings

[0048] Figure 1 It is the Fourier transform infrared spectrum of CE raw material;

[0049] Figure 2 It is the Fourier transform infrared spectrum of EudragitL100 / S100-TPGS grafted product;

[0050] Figure 3 It is the Fourier transform infrared spectrum of the physical mixture of EudragitL100 / S100-TPGS grafted product and CE;

[0051] Figure 4 It is the Fourier transform infrared spectrum of CE SD prepared in Example 5;

[0052] Figure 5 It is the XRD spectrum;

[0053] Figure 6 It is the scanning electron microscope image of CE raw material (300 times);

[0054] Figure 7 It is the scanning electron microscope image of EudragitL100 / S100-TPGS grafted product (300 times);

[0055] Figure 8 It is the scanning electron microscope image of the physical mixture of EudragitL100 / S100-TPGS grafted product and CE (300 times);

[0056] Figure 9 It is the scanning electron microscope image of CE SD prepared in Example 5 (300 times);

[0057] Figure 10 It is a physical appearance diagram of the CE SD prepared in Example 5. Detailed implementation manners

[0058] The following will illustrate the implementation manners of the present invention with reference to the accompanying drawings and preferred embodiments. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It should be understood that the preferred embodiments are only for illustrating the present invention and not for limiting the protection scope of the present invention.

[0059] For those specific technologies or conditions not specified in the specific embodiments, they shall be carried out according to the technologies or conditions described in the literature in this field or according to the product specifications. For the reagents or instruments not indicating the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0060] Example 1

[0061] A preparation method of a CE SD, comprising the following steps:

[0062] S1. Preparation of polyacrylic resin - vitamin E polyethylene glycol 1000 succinate graft:

[0063] S11. Dissolve vitamin E polyethylene glycol 1000 succinate in DMSO, then add EDC (1.2 molar equivalents) and NHS (1.0 molar equivalent), stir at room temperature for 6 h, then rotary evaporate to remove the unreacted EDC / NHS, and freeze-dry to obtain activated vitamin E polyethylene glycol 1000 succinate;

[0064] S12. Add EudragitL100 and EudragitS100 in a mass ratio of 2:1 to a mixed solvent of 70 mL of absolute ethanol and acetone (the volume ratio of absolute ethanol to acetone is 9:1), heat and stir to dissolve at a temperature of 50 °C. After complete dissolution, add the activated TPGS (the amount of activated TPGS used is 1 / 10 of the CE amount), protect under a nitrogen atmosphere, heat and stir to react at a temperature of 50 °C for 48 h, then filter with a 0.22 μm filter membrane, collect the precipitate, and freeze-dry the precipitate to obtain the EudragitL100 / S100-TPGS graft;

[0065] Among them, the reaction equation for preparing the EudragitL100 / S100-TPGS graft is:

[0066] and

[0067] The value of n is a natural number greater than or equal to 1;

[0068] Preparation of S2, CE SD:

[0069] S21. Add the EudragitL100 / S100-TPGS graft copolymer into a 95% anhydrous ethanol-water (v / v) solution, stir magnetically at 50 °C until completely dissolved, then slowly add CE in batches, and then raise the temperature to 60 °C and stir for 30 min. After complete dissolution, a co-solution is obtained. Among them, the mass ratio of the EudragitL100 / S100-TPGS graft copolymer to CE is 67.5:32.5;

[0070] S22. Transfer the co-solution to a preheated distillation flask, evaporate the organic solvent under reduced pressure, quickly place the distillation flask at -20 °C for 24 h to solidify, obtain a block, place the obtained block in a vacuum drying oven for drying, and then crush and sieve to obtain CE SD powder.

[0071] Example 2

[0072] In this example, except that the mass ratio of the EudragitL100 / S100-TPGS graft copolymer to CE is 70:30 and the stirring reaction time in S21 is 45 min, the rest is the same as in Example 1.

[0073] Example 3

[0074] In this example, except that the mass ratio of the EudragitL100 / S100-TPGS graft copolymer to CE is 72.5:27.5 and the stirring reaction time in S21 is 60 min, the rest is the same as in Example 1.

[0075] Example 4

[0076] In this example, except that the mass ratio of EudragitL100 and EudragitS100 is 1:1 and the stirring reaction time in S21 is 45 min, the rest is the same as in Example 1.

[0077] Example 5

[0078] In this example, except that the mass ratio of Eudragit L100 to Eudragit S100 is 1:1 (the mass of Eudragit L100 added in this example is 3.35 g and the mass of Eudragit S100 added is 3.35 g), the mass ratio of the Eudragit L100 / S100-TPGS graft to CE is 70:30 (the mass of the Eudragit L100 / S100-TPGS graft added in this example is 7.0 g, the mass of CE added is 3.0 g, and the same can be obtained for the rest of the examples), and the stirring reaction time in S21 is 60 min, the rest is the same as in Example 1.

[0079] Example 6

[0080] In this example, except that the mass ratio of Eudragit L100 to Eudragit S100 is 1:1 and the mass ratio of the Eudragit L100 / S100-TPGS graft to CE is 72.5:27.5, the rest is the same as in Example 1

[0081] Example 7

[0082] In this example, except that the mass ratio of Eudragit L100 to Eudragit S100 is 1:2 and the stirring reaction time in S21 is 60 min, the rest is the same as in Example 1.

[0083] Example 8

[0084] In this example, except that the mass ratio of Eudragit L100 to Eudragit S100 is 1:2 and the mass ratio of the Eudragit L100 / S100-TPGS graft to CE is 70:30, the rest is the same as in Example 1.

[0085] Example 9

[0086] In this example, except that the mass ratio of Eudragit L100 to Eudragit S100 is 1:2, the mass ratio of the Eudragit L100 / S100-TPGS graft to CE is 72.5:27.5, and the stirring reaction time in S21 is 45 min, the rest is the same as in Example 1.

[0087] Comparative Example 1

[0088] A preparation method of CE-Eudragit L100 / S100 SD comprises the following steps:

[0089] S1. Weigh 3.5 g of Eudragit L100 and 3.5 g of Eudragit S100, add them to an appropriate amount of 95% anhydrous ethanol - water (v / v) solution, and stir magnetically at 50 °C until completely dissolved. Then, slowly add 3.0 g of CE in batches, and continue to stir at 50 °C for 2 h until completely dissolved to obtain a co - solution.

[0090] S2. Quickly transfer the co - solution to a pre - heated distillation flask, evaporate the organic solvent under reduced pressure, and then quickly place the distillation flask at - 20 °C for 24 h to solidify and obtain a block. Place the obtained block in a vacuum drying oven for drying, and then crush and sieve it to obtain CE - Eudragit L100 / S100 SD.

[0091] Comparative Example 2

[0092] A preparation method of ungrafted CE - Eudragit L100 / S100 - TPGS SD includes the following steps:

[0093] S1. Weigh 3.35 g of Eudragit L100 and 3.35 g of Eudragit S100, add them to an appropriate amount of 95% anhydrous ethanol - water (v / v) solution, and stir magnetically at 50 °C until completely dissolved. Then, slowly add 3.0 g of CE in batches, continue to stir at 50 °C for 2 h until completely dissolved, and then add 0.3 g of TPGS and stir at 50 °C to obtain a co - solution.

[0094] S2. Quickly transfer the co - solution to a pre - heated distillation flask, evaporate the organic solvent under reduced pressure, and then quickly place the distillation flask at - 20 °C for 24 h to solidify and obtain a block. Place the obtained block in a vacuum drying oven for drying, and then crush and sieve it to obtain ungrafted CE - Eudragit L100 / S100 - TPGS SD.

[0095] Detection and Analysis

[0096] 1. Dissolution test

[0097] Using hydrochloric acid solution with a pH of 1.2 (simulating gastric juice) and phosphate buffer solution with a pH of 6.8 (simulating intestinal juice) as dissolution media respectively, the dissolution and release of CE SD prepared in Examples 1 - 9, CE-EudragitL100 / S100SD prepared in Comparative Example 1, and CE-EudragitL100 / S100-TPGSSD without grafting treatment prepared in Comparative Example 2 were studied comparatively. Three parallel tests were carried out respectively to calculate the cumulative dissolution rate. The specific operation steps refer to the "Pharmacopoeia of the People's Republic of China" (2020 Edition). The results are shown in Table 1 and Table 2.

[0098] Table 1 Results of dissolution test in acid

[0099]

[0100] Table 2 Results of cumulative dissolution test in phosphate buffer solution

[0101]

[0102] From the results of the dissolution test in acid in Table 1, it can be seen that when EudragitL100 / S100 is used as an enteric coating carrier, whether TPGS is added or not, the dissolution rate of CE SD in hydrochloric acid solution can be effectively reduced. When the drug loading is less than or equal to 30%, the cumulative dissolution rates of each group of CE SD are all less than 10%, meeting the requirement of the Pharmacopoeia of Veterinary Drugs that the dissolution amount of enteric preparations in hydrochloric acid solution within 2 hours is less than 10%. When the drug loading reaches 32.5%, the dissolution rate of CE SD increases significantly, which may be due to the fact that too much CE cannot form a solid dispersion with the carrier, resulting in the premature dissolution of some CE.

[0103] From the results of the dissolution test in phosphate buffer solution in Table 2, it can be known that: 1. The cumulative dissolution rate of Comparative Example 1 without adding the surfactant TPGS was 80.38% at 120 min, and the release of the active ingredient was incomplete; 2. The cumulative dissolution rates of all examples in phosphate buffer solution reached more than 95%, indicating that the enteric coating effect of CE SD prepared by the present invention is very ideal and the drug release is complete; 3. Compared with Comparative Example 2 without grafting treatment, the cumulative dissolution rates of Examples 1 - 9 reached more than 90% at 45 min. Thus, it is proved that CE SD prepared by the present invention can significantly improve the dissolution rate of CE in the intestine, ensure that the drug is completely released before the animal defecates (especially for diarrhea animals), and avoid the loss, waste and environmental pollution of the active ingredient.

[0104] 2. Bitter taste masking test

[0105] Using CE raw materials as the control group and CE SD prepared by the present invention as the experimental group, the bitter taste masking of CE SD prepared in Example 5, CE-EudragitL100 / S100 SD prepared in Comparative Example 1, CE-EudragitL100 / S100-TPGSSD without grafting treatment prepared in Comparative Example 2, and CE raw materials (as the control) was studied comparatively. There were 7 volunteers (4 males and 3 females), aged 26 - 39 years old, with good health, normal taste, no serious oral or gastrointestinal diseases, and none of the females were pregnant or lactating. The volunteers were trained in ascending order of bitter taste intensity. After the training was completed and the assessment was passed, they could enter the formal experiment. The 7 volunteers placed CE SD, CE-EudragitL100 / S100 SD, CE-EudragitL100 / S100-TPGS SD without grafting treatment, and CE raw materials in their mouths and gargled to fully let the tongue feel the taste of the samples, staying in the mouth for 15 s. After testing one sample, they gargled 5 times with pure water until there was no peculiar smell in the mouth. After resting for 10 min, the test of the next sample was carried out. The bitter taste masking effect of the samples was evaluated by scoring. The evaluation index was divided into 5 levels: no bitter taste (0 points), detectable (1 point), slight (2 points), moderate (3 points), and strong bitter taste (4 points). The results are shown in Table 3.

[0106] Table 3 Taste masking effects of different examples

[0107]

[0108] From the test results in Table 3, it can be seen that the bitter taste scores of each group are less than 0.5 points. Thus, it is proved that when EudragitL100 / S100 is used as a combined enteric coating carrier, whether TPGS is added or not, it can effectively mask the bitter taste of CE SD. Unexpectedly, CE SD prepared with the graft has no bitter taste at all, and the taste masking effect is the best. This may be due to the hydrophilic PEG-1000 chain of TPGS and the hydrophobic vitamin E forming an amphiphilic structure, which can self-assemble into micelles or jointly form nanoparticles with EudragitL100 / S100, effectively encapsulating the drug and causing a synergistic effect.

[0109] 3. Characterization of CE SD

[0110] 1) Fourier transform infrared spectroscopy (FTIR) analysis

[0111] The specific operation is as follows: Samples were prepared by the KBr tablet pressing method. The test sample and KBr were mixed evenly at a ratio of 1:50 - 100, and the scanning range was 400 - 4000 cm -1, the infrared spectrum analysis was respectively carried out on the CE raw material, the EudragitL100 / S100-TPGS graft obtained in S1, the physical mixture of the EudragitL100 / S100-TPGS graft and CE (wherein, the physical mixture means that the EudragitL100 / S100-TPGS graft and CE were respectively ground and passed through a 100-mesh medicinal sieve, and then directly mixed evenly, and the same hereinafter), and the CE SD obtained in Example 5. The results are as Figures 1 to 4 shown.

[0112] From Figures 1 to 4 the comparative document, it can be seen that the physical mixture of the EudragitL100 / S100-TPGS graft and CE ( Figure 3 ) shows the superposition of the corresponding infrared spectra of the raw material CE ( Figure 1 ) and the EudragitL100 / S100-TPGS graft ( Figure 2 ). The C-O absorption peak of the CE raw material ( Figure 1 ) and the physical mixture of the EudragitL100 / S100-TPGS graft and CE ( Figure 3 ) at 1140 cm -1 is very obvious, while the vibration absorption peak of the CE SD ( Figure 4 ) obtained in Example 5 at this place has disappeared and has been shielded by the peripheral chemical bonds of the graft. At the same time, the CE raw material ( Figure 1 ) and the physical mixture of the EudragitL100 / S100-TPGS graft and CE ( Figure 3 ) have a typical and strong aromatic ring skeletal vibration absorption peak of CE at 1506 cm -1 , while the vibration absorption peak of the CE SD ( Figure 4 ) obtained in Example 5 at this place has been passivated and disappeared. These results all indicate that the CE raw material and the graft have been combined by hydrogen bonding through C-H and C-O, and CE is dispersed in the graft in an amorphous state.

[0113] 2) X-ray diffraction (XRD) analysis

[0114] The specific operation is as follows: using copper-Kα ray as the radiation source, the radiation intensity is set to 40 kV and 100 mA, the step size is 0.02°, the scanning range is 5~60°, the scanning speed is 4° / min, and the powder X-ray diffraction analysis was respectively carried out on the CE raw material, the EudragitL100 / S100-TPGS graft obtained in S_{1}, the physical mixture of the EudragitL100 / S100-TPGS graft and CE, and the CE SD obtained in Example 5. The results are as Figure 5 shown.

[0115] From Figure 5 Analysis shows that there are four obvious characteristic diffraction peaks of CE raw materials at about 15°, 17°, 18° and 23°. The EudragitL100 / S100-TPGS graft is an amorphous compound without obvious characteristic diffraction peaks. In the physical mixture of EudragitL100 / S100-TPGS graft and CE, partial characteristic diffraction peaks of CE can still be observed at 18° and 23°, indicating that CE still exists in the physical mixture in a crystalline state. In the CE SD prepared in Example 5, the characteristic peaks of CE at the above four positions basically disappear, indicating that the EudragitL100 / S100-TPGS graft has an obvious crystal inhibition effect on CE, and it is once again proved that CE is dispersed in the carrier (graft) in an amorphous state.

[0116] 3) Scanning electron microscopy analysis

[0117] The specific operation is as follows: The CE raw material, the EudragitL100 / S100-TPGS graft prepared in S1, the physical mixture of EudragitL100 / S100-TPGS graft and CE, and the CE SD prepared in Example 5 are respectively collected for images by an electron microscope. Select the area to be observed to collect pictures and observe the specific changes. The results are as Figures 6 to 9 shown.

[0118] From Figures 6 to 9 analysis, it can be seen that the appearance of the CE raw material is irregular crystals with relatively small volume and smooth appearance ( Figure 6 ), while the appearance volume of the EudragitL100 / S100-TPGS graft is relatively large with obvious edges and corners ( Figure 7 ). In the physical mixture of EudragitL100 / S100-TPGS graft and CE, it can be seen that small-volume CE is unevenly distributed around the larger-volume graft ( Figure 8 ), while in the CE SD prepared in Example 5, CE and the EudragitL100 / S100-TPGS graft have fused into a whole, and there is no CE around ( Figure 9 ), and it shows an irregular shape, indicating that CE has been dispersed in the carrier graft in an amorphous state.

[0119] 4) Stability test

[0120] Under the conditions of high temperature (60 °C), high humidity (RH 75% ± 5%), light (4500 ± 500 lx), and accelerated test (placed for 6 months at 40 ± 2 °C and humidity of 75% ± 5%), the stability of CE raw material, CE SD prepared in Example 5, CE-EudragitL100 / S100 SD prepared in Comparative Example 1, and CE-EudragitL100 / S100-TPGS SD without graft treatment prepared in Comparative Example 2 was studied comparatively.

[0121] Among them, the high temperature test was as follows: Take 3 batches each of CE raw material, CE SD prepared in Example 5, CE-EudragitL100 / S100 SD prepared in Comparative Example 1, and CE-EudragitL100 / S100-TPGS SD without graft treatment prepared in Comparative Example 2. Weigh 10.0 g for each batch and spread it flat in a petri dish with a diameter of 10 cm. Place the petri dish in a sealed desiccator, and then put the desiccator into a constant temperature incubator, keep the condition at 60 °C for 10 d, sample and detect at the 0th, 5th, and 10th d respectively. The results are shown in Table 4.

[0122] Table 4 Results of high temperature test

[0123]

[0124] It can be analyzed from Table 4 that the appearance colors of each group are all yellowish-brown powders, no patterns, color spots or other color changes are seen, and no caking is seen either. The degradation rate differences among CE raw material, Comparative Example 1, and Comparative Example 2 are not significant, between 3.21% and 4.57%. However, CE SD in Example 5 prepared by the present invention can effectively reduce the influence of 60 °C high temperature on the active ingredient BBR·Hcl of CE, and the degradation rate is only 0.75%, which is an unexpected gain effect at the beginning of the experimental design.

[0125] The high humidity test was as follows: Take 3 batches each of CE raw material, CE SD prepared in Example 5, CE-EudragitL100 / S100 SD prepared in Comparative Example 1, and CE-EudragitL100 / S100-TPGS SD without graft treatment prepared in Comparative Example 2. Weigh 10.0 g for each batch and spread it flat in a petri dish with a diameter of 10 cm. Take 3 portions for each batch and place them respectively in a desiccator with supersaturated NaCl solution (RH 75% ± 5%) at the bottom, keep the condition at 25 °C for 10 d, sample and detect at the 0th, 5th, and 10th d respectively. The results are shown in Table 5.

[0126] Table 5 Results of high humidity test

[0127]

[0128] Analysis from Table 5 shows that the CE raw material degrades significantly under high humidity conditions, with the appearance showing caking but no obvious discoloration. The degradation rates of the other 3 groups have no significant difference, ranging from 0.61% to 0.75%, and there is no obvious difference in appearance. This indicates that whether TPGS is added to the formulation or not, the CE SD prepared with Eudragit L100 / S100 can well reduce the impact of high humidity (RH 75% ± 5%) on the active ingredient BBR·HCl of CE.

[0129] For the light exposure test, 3 batches of each of the CE raw material, the CE SD prepared in Example 5, the CE-Eudragit L100 / S100 SD prepared in Comparative Example 1, and the un-grafted CE-Eudragit L100 / S100-TPGS SD prepared in Comparative Example 2 were taken. Each batch weighed 10.0 g and was spread flat in a petri dish with a diameter of 10 cm. The petri dish was placed in a light box equipped with fluorescent lamps under the condition of an illuminance of 4500 ± 500 lx for 10 days. Samples were taken and tested on the 0th, 5th, and 10th days respectively, and the results are shown in Table 6.

[0130] Table 6 Results of the light exposure test

[0131]

[0132] Analysis from Table 6 shows that under light exposure conditions, the degradation rates of BBR·HCl in the 4 experimental groups have no significant difference, and there is no caking or discoloration in appearance. This indicates that whether TPGS or Eudragit L100 / S100 is added to the carrier, or whether CE is prepared into SD, the impact of the illuminance of 4500 ± 500 lx on the CE raw material is relatively small.

[0133] For the accelerated test, 3 batches of each of the CE raw material, the CE SD prepared in Example 5, the CE-Eudragit L100 / S100 SD prepared in Comparative Example 1, and the un-grafted CE-Eudragit L100 / S100-TPGS SD prepared in Comparative Example 2 were taken. Each batch weighed 10.0 g and was spread flat in a petri dish with a diameter of 10 cm. It was placed at 40 ± 2°C and a humidity of 75% ± 5% for 6 months. Samples were taken and tested on the 0th, 1st, 2nd, 3rd, and 6th months respectively, and the results are shown in Table 7.

[0134] Table 7 Results of the accelerated test

[0135]

[0136] Note: For the same batch at different time periods, compared with the 0th month, different letters indicate extremely significant differences between groups P < 0.01, and the same letters indicate no significant differences between groups P > 0.05.

[0137] Analysis from Table 7 shows that the active ingredients of the CE raw material degrade significantly, with the appearance showing severe caking, obvious color change, and the emergence of color patches; in Comparative Example 1, the degradation of the active ingredients accelerated starting from the 3rd month. Although there was no caking, there was a slight color change; in Comparative Example 2, the degradation of the active ingredients accelerated starting from the 3rd month, with slight caking and color change occurring; while Example 5 prepared using the present invention is more stable, which can effectively reduce the impact on the active ingredient BBR·Hcl of CE under the conditions of 40±2°C and humidity of 75%±5%. The effective content did not show a significant decrease, and there was no obvious change in appearance. This may be due to the antioxidant property (vitamin E) of TPGS protecting CE from oxidative degradation at high temperatures, and the rigid skeleton of EudragitL100 / S100 enhancing the physical stability of CE. The two work together to cause this, which is a gain effect not anticipated at the beginning of the experimental design.

[0138] 5) Relative bioavailability test

[0139] Taking Comparative Example 1 without added TPGS as the control group, the relative bioavailability of CE SD prepared in Examples 1 - 9 and CE-EudragitL100 / S100-TPGS SD prepared in Comparative Example 2 without graft treatment was studied.

[0140] The specific operation is as follows: Select 110 crossbred pigs of the same parity, similar age, and weight, and randomly divide them into 11 groups with 10 pigs in each group. The pre-feeding period is 7 days, during which all experimental animals are fed with powder without any antibiotics and traditional Chinese medicines. After the pre-feeding period, the corresponding drugs are added to each experimental group. The addition method is a one-time drug administration, and the addition amount is 25 mg / kg·bw (calculated based on the content of the active substance berberine hydrochloride in the Coptis chinensis extract). After drug administration, the blood drug concentration of berberine hydrochloride in plasma is detected at 0h, 0.08h, 0.17h, 0.25h, 0.33h, 0.5h, 0.75h, 1h, 1.5h, 2h, 3h, 4h, 6h, 8h, 12h, 16h, 24h, 36h, 48h, 72h (a total of 20 times). The data is input into the PKSolver pharmacokinetic and pharmacodynamic data processing software for analysis, and the results are shown in Table 8.

[0141] Table 8 Results of relative bioavailability test

[0142]

[0143] The results in Table 8 show that the relative bioavailability of all the examples in pigs is higher than that of Comparative Example 2. This proves that the CE SD treated by graft reaction has a better promoting effect on the absorption process of CE in pigs than the CE-EudragitL100 / S100-TPGS SD without adding TPGS and without graft treatment. This may be because the hydrophilic PEG-1000 chain of TPGS and the hydrophobic vitamin E form an amphiphilic structure, which can self-assemble into micelles or form nanoparticles together with EudragitL100 / S100, reducing the aggregation of CE in the gastrointestinal tract or blood and improving the delivery efficiency. At the same time, after the CE SD wrapped by EudragitL100 / S100-TPGS is released in the intestine, the surface-active effect of TPGS can reduce the surface tension of the mucosa, enhance the drug's penetration through the intestinal epithelium, and thus improve the bioavailability of orally administered CE.

[0144] 6) Clinical treatment effect test for damp-heat diarrhea in piglets (yellow and white scour in piglets)

[0145] A comparative study was conducted on the clinical treatment effects of CE raw material, the CE-EudragitL100 / S100 SD prepared in Comparative Example 1, the un-grafted CE-EudragitL100 / S100-TPGS SD prepared in Comparative Example 2, and the CE SD prepared in Examples 1-9 for damp-heat diarrhea in piglets (yellow and white scour in piglets). From April to June 2024, experiments were carried out in three pig farms in Tongnan District, Wanzhou District and Rongchang District of Chongqing. All experimental animals had natural diseases and were diagnosed after clinical symptom determination and molecular biology identification. Diseased pigs with similar day-old, body weight and disease severity in the same farm were selected as experimental subjects and randomly grouped for the experiment. The blank control group was not given any drug treatment, the control group 1 was given CE raw material treatment, the control group 2 was given the treatment of Comparative Example 1, the control group 3 was given the treatment of Comparative Example 2, and the experimental groups 1-9 were given the treatment of Examples 1-9. Calculated by CE, 5 kg was added to each ton of feed, and the control group and the experimental groups were dosed according to this dosage after converting the effective content. It was used continuously for 3 days, and the results are shown in Table 9.

[0146] Table 9 Treatment effect test for typical cases

[0147]

[0148] As can be seen from the results in Table 9, the clinical symptoms of the pig group were observed every day after medication, and they were divided into effective (including cured) and ineffective (including dead). After the medication ended, if the symptoms of the diseased pigs completely disappeared and no new diarrhea symptoms appeared within 14 days after drug withdrawal, it was judged as effective; if the diseased pigs still had obvious clinical symptoms or relapsed diarrhea symptoms within 14 days after drug withdrawal, it was judged as ineffective. The results showed (Table 9) that the CE SD prepared by the present invention had significantly better clinical treatment effects when used for damp-heat diarrhea in piglets (yellow and white scour in piglets).

[0149] 7) Physical appearance

[0150] Figure 10 It is a physical diagram of the CE SD prepared in Example 5. As can be observed from Figure 10 it, the CE SD prepared in Example 5 is a yellow powder, so it will not release bitterness due to animal chewing, has good palatability, stable properties, good uniformity, has good fluidity, is convenient for mixing with feed for administration, and has potential new drug development value in the technical field of veterinary drug preparations.

[0151] In summary, the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft provided by the present invention can form an amphiphilic structure by utilizing the hydrophilic PEG-1000 chain of TPGS and the hydrophobic vitamin E, and can self-assemble into micelles or jointly form nanoparticles with Eudragit L100 / S100, so that as a carrier, it can effectively encapsulate the active pharmaceutical ingredient CE, reduce the aggregation of CE in the gastrointestinal tract or blood, and improve the delivery efficiency. At the same time, after the Eudragit L100 / S100-TPGS encapsulated CE SD is released in the intestine, the surface-active effect of TPGS can reduce the surface tension of the mucosa, enhance the intestinal epithelial penetration of the drug, and thus improve the bioavailability of orally administered CE. At the same time, by utilizing the antioxidant property (vitamin E) of TPGS to protect the drug CE from oxidative degradation, the rigid skeleton of Eudragit L100 / S100 can also enhance the physical stability of CE and effectively extend the storage time. As an excipient approved by the FDA, TPGS has high biocompatibility, can neutralize the possible mucosal irritation caused by orally administered Eudragit L100 / S100, and improve the drug safety.

[0152] The preparation method of the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft provided by the present invention is to carry out an esterification reaction between the carboxyl groups of Eudragit L100 / S100 and the hydroxyl groups of TPGS to form an ester bond, thereby grafting TPGS onto Eudragit L100 / S100, and innovatively preparing the graft Eudragit L100 / S100-TPGS. The enteric-coated CE SD prepared with this graft avoids the significant solubilization effect of the surfactant TPGS on CE in a pH 1.2 hydrochloric acid solution, and at the same time increases its cumulative dissolution rate in a pH 6.8 phosphate buffer solution, resulting in a low dissolution rate of CE in the stomach and rapid disintegration and dissolution in the intestine. Moreover, the preparation method has a simple process, is easy to operate, has a low cost, and the required equipment is conventional equipment in veterinary drug factories, facilitating large-scale industrial production, having great economic and social benefits, and meeting the requirements of green ecology and sustainable development.

[0153] The preparation method of the CE SD of the present invention is to prepare an enteric-coated CE SD by the solvent method, without the need to heat both the drug and the carrier to the molten state, avoiding the reduction of the efficacy of the active ingredient BBR·HCl of CE due to thermal instability, and ensuring the drug efficacy. Moreover, the preparation method is simple, suitable for large-scale production, effectively masks the bitterness of CE, allows CE to pass through the stomach and intestine, solves the problem of gastric irritation caused by the dissolution of oral CE in the stomach, and the prepared CE SD is not easily degraded and deteriorated, has a longer shelf life, higher bioavailability, and better clinical treatment effect.

[0154] The CE SD provided by the present invention, in addition to the raw material drug CE, the carrier is only a graft prepared from an enteric material and a surfactant, has a high drug loading, and on the basis of no coating layer structure, simultaneously overcomes the disadvantages of poor palatability and large gastric irritation of CE. Moreover, the CE SD is in powder form, will not release bitterness due to animal chewing, has good palatability, stable properties, good uniformity, has good fluidity, is convenient for mixing with feed for administration, and has potential new drug development value in the field of veterinary drug preparation technology.

[0155] The above embodiments are only preferred embodiments given to fully illustrate the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or transformations made by those skilled in the art on the basis of the present invention are all within the protection scope of the present invention.

[0156] In addition, it should be understood that although this specification is described according to embodiments, not each embodiment only contains an independent technical solution. This narrative way of the specification is only for clarity. Those skilled in the art should regard the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A preparation method of a polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft, characterized in that, It includes the following steps: Dissolve vitamin E polyethylene glycol 1000 succinate in dimethyl sulfoxide, then add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide to obtain activated vitamin E polyethylene glycol 1000 succinate; Add Eudragit L100 and Eudragit S100 to a mixed solvent of absolute ethanol and acetone, heat to dissolve, then add the activated vitamin E polyethylene glycol 1000 succinate, heat and react under an inert atmosphere, filter to obtain a polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft; The molar ratio of the 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride to the N-hydroxysuccinimide is 1.2:1; The mass ratio of the Eudragit L100, Eudragit S100 and the activated vitamin E polyethylene glycol 1000 succinate is 2:1:0.08 to 1:2:0.

12.

2. The preparation method of the polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft according to claim 1, characterized in that, The temperature for heating and dissolving is 45-55°C; And / or, the temperature for heating and reacting under an inert atmosphere is 48-52°C, and the time is 45-50 h.

3. A polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft prepared by the preparation method according to claim 1 or claim 2.

4. Use of the polyacrylic acid resin-vitamin E polyethylene glycol 1000 succinate graft according to claim 3, characterized in that, The polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft is used as a drug carrier.

5. The application according to claim 4, wherein The polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft is used as an enteric-coated carrier.

6. A solid dispersion of Coptis chinensis extract, characterized in that, It includes the Coptis chinensis extract and the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft according to claim 3.

7. The preparation method of the solid dispersion of the Coptis chinensis extract according to claim 6, characterized in that, It includes the following steps: Add the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft to an ethanol aqueous solution, after heat treatment, add the Coptis chinensis extract in batches, then carry out heat-up treatment, low-temperature distillation, solidification, and drying to obtain a solid dispersion of the Coptis chinensis extract.

8. The preparation method of the solid dispersion of Coptis chinensis extract according to claim 7, characterized in that, The temperature for the heat treatment is 48-52°C; And / or, the temperature for the heat-up treatment is 58-62°C; And / or, the temperature for the low-temperature distillation is less than 60°C; And / or, the mass ratio of the polyacrylic resin-vitamin E polyethylene glycol 1000 succinate graft to the Coptis chinensis extract is 6-8:2-4; And / or, the solidification is to place the residue after distillation at a temperature of -25 to -15°C for solidification for 20-28 h; And / or, the drying is to dry the solidified block under vacuum conditions.

9. Use of a solid dispersion of the Coptis chinensis extract prepared by the preparation method according to claim 7 or claim 8 in a drug for preventing and treating animal diarrhea diseases.

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