Composition containing prednisolone and preparation method and application thereof

Abstract

The present application belongs to the field of pharmaceutical preparations, and provides a composition containing prednisolone, a preparation method thereof, and an application thereof, wherein the composition containing prednisolone comprises a matrix and prednisolone dispersed in the matrix; the matrix is ​​a copolymer of citric acid and methacrylic acid; the methacrylic acid copolymer is a methacrylic acid copolymer type A and a methacrylic acid copolymer type B. The composition of the present invention can be released at a specific point in the colon, greatly improving the effect of the drug in treating lesions in the colon.

Description

Field of the Invention

[0001] The present application belongs to the field of pharmaceutical preparations. Specifically, the present application provides a composition containing prednisolone and a preparation method and application thereof. Background Art

[0002] Prednisolone is a synthetic corticosteroid drug, belonging to a potent glucocorticoid drug, which has anti-inflammatory, anti-allergic and immunosuppressive effects, can inhibit the proliferation of connective tissue, reduce the permeability of capillary walls and cell membranes, reduce inflammatory exudation, and can inhibit the formation and release of histamine and other toxic substances. When severely toxic infection occurs, it can be used in conjunction with a large amount of antibacterial drugs to have good cooling, anti-toxic, anti-inflammatory, anti-shock and symptom relief effects. For example, it can treat ulcerative colitis, which is a chronic inflammatory bowel disease, in which the large intestine (colon) is inflamed and ulcerated (with dents or erosions), resulting in multiple disease attacks characterized by bloody diarrhea, abdominal cramps and fever. The long-term risk of ulcerative colitis patients suffering from colon cancer is higher than that of people without ulcerative colitis. The local administration method for treating ulcerative colitis in the prior art is generally the method of enema and suppository, while the oral administration method can be released in the stomach or the front end of the intestine, and it is impossible or rarely released at a fixed point in the colon, and the effect is greatly reduced. Summary of the invention

[0003] In order to overcome the above problems existing in the prior art, the present application provides a prednisolone-containing composition and a preparation method and application thereof.

[0004] In a first aspect, the present application provides a composition containing prednisolone, comprising a matrix and prednisolone dispersed in the matrix; wherein the matrix is ​​a copolymer of citric acid and methacrylic acid; and the methacrylic acid copolymer is methacrylic acid copolymer type A and methacrylic acid copolymer type B.

[0005] Furthermore, the methacrylic acid copolymer type A is a copolymer in which the molar ratio of methacrylic acid to methyl methacrylate is 1:1, and the methacrylic acid copolymer type B is a copolymer in which the molar ratio of methacrylic acid to methyl methacrylate is 1:2.

[0006] Furthermore, the weight ratio of prednisolone to matrix in the composition is 1:1-10, and can be 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10 or a range formed by any two of the above values ​​and a value within the range.

[0007] Furthermore, the weight ratio of the citric acid to methacrylic acid copolymer is 1:10-100, and can be 1:10, 1:20, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, 1:100 or a range formed by any two of the above values ​​and a value within the range.

[0008] Furthermore, the weight ratio of the methacrylic acid copolymer type A to the methacrylic acid copolymer type B is 1:1-5, and can be 1:1, 1:2, 1:3, 1:4, 1:5 or a range formed by any two of the above values ​​and a value within the range.

[0009] Furthermore, the composition is a microsphere, and the particle size of the microsphere is 100-700 μm, which can be 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm or a range formed by any two of the above values ​​and a value within the range.

[0010] In a second aspect, the present application provides a method for preparing a composition containing prednisolone, the method comprising: (1) mixing prednisolone and a matrix in the presence of an organic solvent; (2) adding the mixed product of step (1) into water in the presence of an emulsifier for molding; wherein the matrix is ​​a copolymer of citric acid and methacrylic acid; and the methacrylic acid copolymer is methacrylic acid copolymer type A and methacrylic acid copolymer type B.

[0011] Furthermore, the content of prednisolone in the mixed system of step (1) is 0.5-10% by weight, and can be 0.5% by weight, 1% by weight, 2% by weight, 3% by weight, 4% by weight, 5% by weight, 6% by weight, 7% by weight, 8% by weight, 9% by weight, 10% by weight, or a range formed by any two of the above values, or a value within the range.

[0012] Furthermore, the dosage ratio of prednisolone to matrix is ​​1:1-10, and can be 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10 or a range formed by any two of the above values ​​and a value within the range.

[0013] Furthermore, the usage ratio of the citric acid to methacrylic acid copolymer is 1:10-100, which may be 1:10, 1:20, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, 1:100 or a range formed by any two of the above values ​​and a value within the range.

[0014] Furthermore, the usage ratio of the methacrylic acid copolymer type A to the methacrylic acid copolymer type B is 1:1-5, and can be 1:1, 1:2, 1:3, 1:4, 1:5 or a range formed by any two of the above values ​​and a value within the range.

[0015] Furthermore, the weight ratio of the emulsifier to water in step (2) is 1:10-1000, and can be 1:10, 1:50, 1:100, 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, 1:1000 or a range formed by any two of the above values ​​and a value within the range.

[0016] Furthermore, the mixing conditions include: a temperature of 0-30°C, which can be 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C or a range formed by any two of the above values ​​and a value within the range; a time of 0.1-3h, which can be 0.1h, 0.5h, 1h, 1.5h, 2h, 2.5h, 3h or a range formed by any two of the above values ​​and a value within the range.

[0017] Furthermore, the organic solvent is not limited and can dissolve prednisolone and the matrix. Preferably, the organic solvent is selected from at least one of dichloromethane, 1,4-dioxane, ethyl formate, ethyl acetate, methyl formate, methyl acetate, butyl acetate, isoamyl acetate, methyl isobutyl ketone, methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, tert-butanol, ethylene glycol and glycerol.

[0018] Furthermore, the emulsifier is selected from at least one of sodium dodecyl sulfate, Tween 20, Tween 80, Span 60, Span 80, polyvinyl alcohol, sucrose fatty acid ester and poloxamer, preferably at least one of Tween 80, sodium dodecyl sulfate and polyvinyl alcohol.

[0019] Furthermore, the method also includes performing solid-liquid separation on the product obtained in step (2).

[0020] Furthermore, the method also includes washing the solid obtained by solid-liquid separation.

[0021] In a third aspect, the present application provides a composition containing prednisolone prepared by the above method.

[0022] In the present invention, prednisone and each matrix used in the above method are basically all prepared into a composition, and the loss is very small and can be ignored. Therefore, the content of each component in the composition has been described above and will not be repeated here.

[0023] In a fourth aspect, the present application provides the use of the above-mentioned prednisolone-containing composition in the preparation of a pharmaceutical preparation.

[0024] The drug is prednisolone.

[0025] In the present invention, the pharmaceutical preparation is prepared into at least one of a dry suspension, a tablet, a capsule, a dispersible tablet and an orally disintegrating tablet by adding different pharmaceutical excipients.

[0026] In a fifth aspect, the present application provides the use of the above-mentioned prednisolone-containing composition in the preparation of a drug or reagent for reducing the content of sIgA (secretory immunoglobulin A) and / or IL-2 (interleukin-2) in the intestinal mucus of animals.

[0027] In a sixth aspect, the present application provides the use of citric acid and methacrylic acid copolymers in the preparation of colon-directed release drugs, wherein the methacrylic acid copolymers are methacrylic acid copolymer type A and methacrylic acid copolymer type B.

[0028] The drug is prednisolone.

[0029] The citric acid described in the present application is preferably unhydrated citric acid. Citric acid hydrates, such as citric acid monohydrate, citric acid dihydrate, etc., may also be used without affecting the formation of the oil phase. Those skilled in the art can convert the dosage according to the molecular formula.

[0030] Through the above technical scheme, the composition of the present invention can be released at a fixed point in the colon, greatly improving the effect of the drug in treating lesions in the colon. The colon fixed point release drug refers to a preparation that is basically not released in the upper gastrointestinal tract and is mostly or completely released in the colon, that is, it is not released (0%) in a specified acidic medium and is not released or is almost not released (less than 10%) in a pH 6.8 phosphate buffer within a certain period of time, but is mostly or completely released in a pH 7.5-8.0 phosphate buffer within a required time.

[0031] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 is the result of determination of inflammatory factor sIgA in intestinal mucus of mice in each group;

[0033] Figure 2 is the result of determination of inflammatory factor IL-2 in intestinal mucus of mice in each group;

[0034] Figure 3 are the pathological results of the distal colon of mice in each group (the left column is 10×10 times, and the right column is 10×40 times);

[0035] Figure 4 These are the rectal pathological results of mice in each group (the left column is 10×10 times, and the right column is 10×40 times). DETAILED DESCRIPTION

[0036] Methacrylic acid copolymer type A (copolymer of methacrylic acid and methyl methacrylate in a molar ratio of 1:1) Source: Evonik Operations GmbH;

[0037] Methacrylic acid copolymer type B (copolymer of methacrylic acid and methyl methacrylate in a molar ratio of 1:2) Source: Evonik Operations GmbH;

[0038] Prednisolone source: Henan Lihua Pharmaceutical Co., Ltd.;

[0039] Example 1

[0040] (1) dissolving prednisolone and a matrix (a weight ratio of citric acid to methacrylic acid copolymer of 1:50, a weight ratio of methacrylic acid copolymer type A to methacrylic acid copolymer type B of 1:3) in dichloromethane at a weight ratio of 1:5 to form an oil phase, wherein the content of prednisolone in the oil phase is 2%, the temperature is 20° C., and the time is 2 h;

[0041] (2) mixing polyvinyl alcohol and water in a weight ratio of 1:200 to form an aqueous phase;

[0042] (3) adding the oil phase in step (1) to the water phase in step (2) and stirring;

[0043] (4) The stirred product in step (3) is subjected to solid-liquid separation to obtain microspheres with a particle size of 100-700 μm.

[0044] Example 2

[0045] (1) dissolving prednisolone and a matrix (the weight ratio of citric acid and methacrylic acid copolymer is 1:50, and the weight ratio of methacrylic acid copolymer type A and methacrylic acid copolymer type B is 1:3) in a solution of ethyl acetate and ethanol (the weight ratio of ethyl acetate to ethanol is 15:1) at a weight ratio of 1:5 to form an oil phase, wherein the content of prednisolone in the oil phase is 1%, the temperature is 30°C, and the time is 1 hour;

[0046] (2) mixing sodium lauryl sulfate and water in a weight ratio of 1:50 to form an aqueous phase;

[0047] (3) adding the oil phase in step (1) to the water phase in step (2) and stirring;

[0048] (4) The stirred product in step (3) is subjected to solid-liquid separation to obtain microspheres with a particle size of 100-700 μm.

[0049] Example 3

[0050] (1) dissolving prednisolone and a matrix (the weight ratio of citric acid and methacrylic acid copolymer is 1:50, and the weight ratio of methacrylic acid copolymer type A and methacrylic acid copolymer type B is 1:3) in ethyl formic acid at a weight ratio of 1:5 to form an oil phase, wherein the content of prednisolone in the oil phase is 5%, the temperature is 0°C, and the time is 3 hours;

[0051] (2) mixing Tween 80 and water at a weight ratio of 1:100 to form an aqueous phase;

[0052] (3) adding the oil phase in step (1) to the water phase in step (2) and stirring;

[0053] (4) The stirred product in step (3) is subjected to solid-liquid separation to obtain microspheres with a particle size of 100-700 μm.

[0054] Example 4

[0055] The method of Example 1 was followed, except that the weight ratio of citric acid to methacrylic acid copolymer was 1:10. Microspheres with a particle size of 100-700 μm were obtained.

[0056] Example 5

[0057] The method of Example 1 was followed, except that the weight ratio of citric acid to methacrylic acid copolymer was 1:100. Microspheres with a particle size of 100-700 μm were obtained.

[0058] Example 6

[0059] The method of Example 1 was followed, except that the weight ratio of methacrylic acid copolymer type A to methacrylic acid copolymer type B was 1:1. Microspheres with a particle size of 100-700 μm were obtained.

[0060] Example 7

[0061] The method of Example 1 was followed, except that the weight ratio of methacrylic acid copolymer type A to methacrylic acid copolymer type B was 1:5. Microspheres with a particle size of 100-700 μm were obtained.

[0062] Example 8

[0063] The method of Example 1 is followed, except that the weight ratio of prednisolone to matrix is ​​1:10. Microspheres with a particle size of 100-700 μm are obtained.

[0064] Example 9

[0065] The method of Example 1 is followed, except that the weight ratio of prednisolone to matrix is ​​1:1. Microspheres with a particle size of 100-700 μm are obtained.

[0066] Comparative Example 1

[0067] The method of Example 1 was followed, except that citric acid was not added and the matrix was methacrylic acid copolymer type A and methacrylic acid copolymer type B. Microspheres with a particle size of 100-700 μm were obtained.

[0068] Comparative Example 2

[0069] The method of Example 1 was followed, except that methacrylic acid copolymer type A was not added and the matrix was citric acid and methacrylic acid copolymer type B. Microspheres with a particle size of 100-700 μm were obtained.

[0070] Comparative Example 3

[0071] The method of Example 1 was followed, except that methacrylic acid copolymer type B was not added and the matrix was citric acid and methacrylic acid copolymer type A. Microspheres with a particle size of 100-700 μm were obtained.

[0072] Comparative Example 4

[0073] The method of Example 1 was followed, except that methacrylic acid copolymer type A and methacrylic acid copolymer type B were not added, and the matrix was citric acid, to obtain microspheres with a particle size of 100-700 μm.

[0074] Comparative Example 5

[0075] The method of Example 1 was followed, except that citric acid was replaced by sodium citrate, and the matrix was sodium citrate, methacrylic acid copolymer type A, and methacrylic acid copolymer type B. Microspheres with a particle size of 100-700 μm were obtained.

[0076] Test Case

[0077] 1. In vitro dissolution

[0078] Method: The above examples and comparative examples were measured according to the dissolution and release determination method (Chinese Pharmacopoeia 2020 General Rules 0931 Second Method). The dissolution conditions are as follows:

[0079] Test method in acid: Use 500ml of 0.1mol / L hydrochloric acid solution as dissolution medium, rotate at 50 revolutions per minute, operate according to the law, take samples after 2 hours, and replenish with an equal volume of 0.1mol / L hydrochloric acid solution at the same time. The limit shall not be detected. The results are shown in Table 1.

[0080] Small Intestine Enteric Coating Test Method: Use pH 6.8 phosphate buffer as dissolution medium, rotate at 50 revolutions per minute, operate according to the law, take samples after 3 hours, and replenish with an equal volume of pH 6.8 phosphate buffer at the same time. The limit shall not exceed 10% of the feed amount. The results are shown in Table 1.

[0081] Colon dissolution test method: Use pH 7.5 phosphate buffer as dissolution medium, rotate at 50 revolutions per minute, operate according to the law, take samples after 30 minutes, and replenish with an equal volume of pH 7.5 phosphate buffer at the same time. The limit is not less than 70% of the feed amount. The results are shown in Table 1.

[0082] Table 1 Dissolution test results

[0083]

[0084] As shown in Table 1, the prednisolone in the composition of the present invention did not dissolve in acid for 2 hours (0%), dissolved less in pH 6.8 medium for 3 hours (less than 10%), and was almost completely dissolved in 7.5-8.0 medium in 120 minutes.

[0085] 2. Pharmacodynamics study

[0086] (1) Test method: Dextran Sulfate Sodium Salt (DSS, MW: 36000-50000) was used to establish the ulcerative colitis (UC) model. The acute colitis model can be established by giving animals free access to a 4% DSS (MW: 36000-50000) aqueous solution. The symptoms of this model are very similar to those of human UC, mainly manifested as diarrhea, mucous stools, fecal occult blood, gross bloody stools, weight loss, decreased activity, and poor coat color.

[0087] (2) After 1 week of adaptive feeding, mice (weight 20-30 g) were randomly divided into groups, with 10 mice in each group. Except for the blank control group, the mice to be modeled drank 1 mL / day of 4% DSS aqueous solution on days 1, 3, 5, and 8. The blank control group was given an equal amount of distilled water (fasting state).

[0088] After successful modeling (the day after the last drinking of DSS aqueous solution), the mice in the model control group were given distilled water by gavage at 1 mL / kg. The mice in the positive drug group were given the drug solution (wherein the prednisolone content in the drug solution was 13 mg / mL) by gavage at 1 mL / kg, and the mice in Example 1, Example 2, Comparative Example 1, and Comparative Example 5 were given once / day / mouse for 7 consecutive days. The blank control group was given an equal amount of distilled water throughout the experiment.

[0089] 12 hours after the last administration of the drug, the mice in each group were killed and samples were collected. The ELISA method was used to detect the content of sIgA and IL-2 in intestinal mucus. The results are shown in Table 2. Figure 1 and Figure 2 .

[0090] Table 2 Contents of sIgA and IL-2

[0091] Group sIgA (μg / ml) IL-2 (ng / ml) Blank control group 38±5 48±6 Model Group 130±11 162±12 Example 1 50±4 55±4 Example 2 55±5 60±5 Comparative Example 1 101±10 110±11 Comparative Example 5 98±6 106±7

[0092] Through Table 2, Figure 1 and Figure 2 It can be seen that compared with the control group, the content of sIgA and IL-2 in the intestinal mucus of the mice in the model group increased significantly, and the difference was statistically significant (P < 0.05). Compared with the model group, the content of the two indicators of the mice in Example 1 and Example 2 groups decreased significantly, and the difference was statistically significant (P < 0.05), and the content of the two indicators of the mice in Comparative Example 1 and Comparative Example 5 groups decreased less. This shows that the embodiments of the present invention have significant effects.

[0093] (3) Twelve hours after the last administration, the mice were killed and dissected, and the distal colon and rectal tissues were separated and fixed in 4% paraformaldehyde. HE staining was performed, and the pathological changes of the distal colon and rectal tissues were observed under a light microscope. The results are shown in the figure. Figure 3 and Figure 4 , the left side is observed at 10×10 times, and the right side is observed at 10×40 times.

[0094] The results showed that in the blank control group, the tissue structures of the rectum and distal colon of mice were complete and clear, the thickness of each layer was normal, the colon glands were neatly arranged, the number was abundant, the number of goblet cells was abundant, there was no detached mucosal tissue in the intestine, and the content of inflammatory factors was low. The colon tissue structure of mice in the model group was completely destroyed, the number of colon glands was greatly reduced, and even some colon gland structures were invisible, the number of goblet cells was small, the rectal tissue was swollen, and the mucosal tissue was often shed in the intestine, and a large number of inflammatory cytokines were gathered. After treatment with the samples of the embodiments of the present invention, the tissue structure damage was significantly improved, the colon gland structure was relatively clear, neatly arranged, abundant in number, normal in thickness, and the number of goblet cells was abundant. The mucosal tissue shed in the intestine was reduced, and the inflammatory cytokines were basically cleared, which was basically consistent with the blank control animal tissue. Among them, the colon gland goblet cells of Examples 1 and 2 were significantly more than those of Comparative Examples 1 and 5. It shows that the therapeutic effects of Examples 1 and 2 are better than those of Comparative Examples 1 and 5.

[0095] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. A composition containing prednisolone, characterized in that The composition comprises a matrix and prednisolone dispersed in the matrix, wherein the weight ratio of prednisolone to the matrix in the composition is 1:1-10; the matrix is ​​a copolymer of citric acid and methacrylic acid, wherein the weight ratio of the citric acid and methacrylic acid copolymer is 1:10-100; the methacrylic acid copolymer is a methacrylic acid copolymer type A and a methacrylic acid copolymer type B; and the weight ratio of the methacrylic acid copolymer type A to the methacrylic acid copolymer type B is 1:1-5; The composition is microspheres, and the particle size of the microspheres is 100-700 μm.

2. A method for preparing a composition containing prednisolone, characterized in that: The method comprises: (1) mixing prednisolone and a matrix in the presence of an organic solvent; the content of prednisolone in the mixed system of step (1) is 0.5-10% by weight; the matrix is ​​a copolymer of citric acid and methacrylic acid; the dosage ratio of prednisolone to the matrix is ​​1:1-10; the dosage ratio of citric acid to methyl copolymer is 1:10-100; the methacrylic acid copolymer is a methacrylic acid copolymer type A and a methacrylic acid copolymer type B; the dosage ratio of the methacrylic acid copolymer type A to the methacrylic acid copolymer type B is 1:1-5; (2) in the presence of an emulsifier, adding the mixed product of step (1) into water for molding.

3. The method according to claim 2, wherein: The weight ratio of the emulsifier to water in step (2) is 1:10-1000; And / or, the mixing conditions include: temperature of 0-30° C. and time of 0.1-3 h.

4. The method according to claim 3, wherein: The organic solvent is selected from at least one of dichloromethane, 1,4-dioxane, ethyl formate, ethyl acetate, methyl formate, methyl acetate, butyl acetate, isoamyl acetate, methyl isobutyl ketone, methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, tert-butanol, ethylene glycol and glycerol; and / or, the emulsifier is at least one selected from the group consisting of sodium dodecyl sulfate, Tween 20, Tween 80, Span 60, Span 80, polyvinyl alcohol, sucrose fatty acid esters and poloxamer; And / or, the method further comprises performing solid-liquid separation on the product obtained in step (2).

5. The prednisolone-containing composition prepared by the method according to any one of claims 2 to 4.

6. Use of the prednisolone-containing composition according to claim 1 or 5 in the preparation of a pharmaceutical preparation.

7. Use of the prednisolone-containing composition according to any one of claims 1 or 5 in reducing the content of sIgA and / or IL-2 in intestinal mucus of animals.

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