Liposome containing active ingredient celecoxib, preparation and application

Abstract

The invention belongs to the technical field of specific therapeutic activity of pharmaceutical preparations, and relates to a liposome containing an active component celecoxib, a preparation and application. The liposome disclosed by the invention is prepared by adopting a film dispersion method and is prepared from celecoxib, sauchinone, phospholipid, cholesterol and a polyvinyl alcohol-sodium alginate blend. Celecoxib and sauchinone are wrapped by lipidosome, the water solubility of the lipidosome is improved, and the drug loading capacity and the encapsulation efficiency of the lipidosome are higher through a specific formula and a specific method; in addition, the combined use of the celecoxib and sauchinone has a synergistic effect, and the effect of treating rheumatoid arthritis is superior to that of single use of the celecoxib.

Description

Technical Field

[0001] The invention belongs to the technical field of specific therapeutic activity of pharmaceutical preparations, and specifically relates to a liposome containing the active ingredient celecoxib, a preparation and a use thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not be necessarily regarded as an admission or any form of suggestion that the information constitutes the prior art already known to those skilled in the art.

[0003] Celecoxib, as a nonsteroidal anti-inflammatory drug (NSAIDs), is mainly used clinically to relieve the symptoms and signs of osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, and to treat acute pain in adults. It achieves anti-inflammatory, analgesic and antipyretic effects by selectively inhibiting the activity of cyclooxygenase-2 (COX-2) and reducing the synthesis of prostaglandins. Compared with traditional nonsteroidal anti-inflammatory drugs, celecoxib has relatively few adverse reactions to the gastrointestinal tract. However, celecoxib still has some limitations in clinical application. First, celecoxib has poor water solubility and is almost insoluble in water, which leads to its low oral bioavailability. The dissolution of drugs in the gastrointestinal tract is a prerequisite for absorption. Because celecoxib is poorly soluble, its dissolution rate in the gastrointestinal tract is slow and its absorption is incomplete, which affects the efficacy of the drug. Secondly, the half-life of celecoxib is relatively short. In order to maintain effective blood drug concentrations, frequent administration is required, which not only brings inconvenience to patients, but may also cause large fluctuations in blood drug concentrations and increase the risk of adverse reactions, such as adverse reactions of the cardiovascular system.

[0004] Sauropodazone (CAS: 177931-17-8) is a natural plant-derived anti-inflammatory and antioxidant compound belonging to the anthocyanidin class of compounds, which has multiple biological activities such as anti-tumor, anti-inflammatory, and antioxidant. Studies have shown that Sauropodazone can regulate the immune system, reduce inflammatory responses, and inhibit the proliferation of tumor cells to a certain extent, and has potential clinical application value. However, Sauropodazone has poor water solubility, resulting in low oral bioavailability and difficulty in fully exerting its therapeutic effect.

[0005] In order to solve the above problems, liposome, as an advanced drug delivery system, has gradually become an important tool for improving drug solubility, increasing drug bioavailability and prolonging drug action time. Liposome is a tiny lipid ball composed of phospholipid molecules, which can effectively encapsulate hydrophobic and water-soluble drugs, thereby improving the stability and bioavailability of drugs. In addition, liposome has the advantages of good biocompatibility, easy preparation, high drug loading, etc., and can realize sustained release or targeted delivery of drugs in the process of drug release, and enhance drug treatment effect. However, the research on liposome preparations containing celecoxib and saururacetam is relatively less. Summary of the invention

[0006] The invention overcomes the shortcomings of the prior art and provides a liposome containing active ingredients celecoxib and sauropodazone. The liposome containing active ingredients celecoxib and sauropodazone is composed of 1 to 5 parts by weight of celecoxib, 0.3 to 0.8 parts by weight of sauropodazone, 4 to 8 parts by weight of phospholipids, 2 to 4 parts by weight of cholesterol, and 0.2 to 0.6 parts by weight of a polyvinyl alcohol-sodium alginate blend.

[0007] The celecoxib structural formula is as follows: .

[0008] The structural formula of the saururaceous ketone is as follows: .

[0009] The polyvinyl alcohol-sodium alginate blend is prepared by mixing polyvinyl alcohol and sodium alginate in a mass ratio of 1:1.

[0010] Furthermore, the phospholipid is egg yolk phospholipid and / or hydrogenated soybean lecithin.

[0011] The present invention also provides a method for preparing the liposome containing the active ingredients celecoxib and saururusin, which belongs to a thin film dispersion method and is specifically as follows: (1) Dissolve celecoxib and saururus in an organic solvent and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) dissolving the polyvinyl alcohol-sodium alginate blend in a buffer solution and preheating it at 40°C to 50°C, and adding it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes are homogenized under high pressure to obtain liposomes containing the active ingredients celecoxib and trichloroacetic acid.

[0012] Furthermore, the organic solvent is chloroform and / or ethanol; the buffer solution is selected from one of citrate buffer, phosphate buffer and acetate buffer; the pH of the buffer solution is 5.5-6.5; the pressure of the high-pressure homogenization is 200-800 bar, and the cycle is 2-5 times.

[0013] The present invention also provides an oral preparation, which is composed of the above-mentioned liposome containing active ingredients celecoxib and saururacetam and pharmaceutically acceptable excipients. The oral preparation is a tablet, a granule, a capsule, or an oral liquid. The excipients are a diluent, a wetting agent, a binder, a disintegrant, a lubricant, a solubilizing agent, a flavoring agent, a preservative, and a coloring agent.

[0014] The tablets are prepared by taking the above-mentioned liposomes containing the active ingredients celecoxib and trichloroacetic acid, freeze-drying them into powder, adding one or more of starch, mannitol, dextrin, lactose, and microcrystalline cellulose, adding one or more of sodium carboxymethyl starch (CMS-Na), cross-linked polyvinylpyrrolidone (PVPP), cross-linked sodium carboxymethyl cellulose (CCMC-Na), and low-substituted hydroxypropyl cellulose, adding one or more of magnesium stearate, micropowdered silica gel, talc, and polyethylene glycol, and pressing the tablets to obtain the tablets. The tablets can also be prepared by wet granulation: the liposomes containing the active ingredients celecoxib and saururacetam are taken, concentrated into a paste with a relative density of 1.4 to 1.6, and one or more of starch, mannitol, dextrin, lactose, and microcrystalline cellulose are added, and one or more of sodium carboxymethyl starch (CMS-Na), cross-linked polyvinylpyrrolidone (PVPP), cross-linked sodium carboxymethyl cellulose (CCMC-Na), and low-substituted hydroxypropyl cellulose are added, wet granulated, granulated, one or more of magnesium stearate, micro-powdered silica gel, talc, and polyethylene glycol are added, and tablets are obtained.

[0015] The preparation of the oral liquid is as follows: taking the above-mentioned liposomes containing the active ingredients celecoxib and sauropodium pyrrolidone, adding one or more of ethanol, propylene glycol, polysorbate 80, and poloxamer, adding one or more of sucrose, stevioside, aspartame, and flavors, adding one or more of parabens (parahydroxybenzoates), benzoic acid and its sodium salt, sorbic acid and its potassium salt, and benzalkonium bromide, and mixing to obtain the oral liquid.

[0016] The present invention also provides the use of the oral preparation containing the active ingredients celecoxib and sauropodii ketone in preparing a drug for treating arthritis.

[0017] Compared with the prior art, the technical effects of the present invention are: (1) Improving drug solubility and bioavailability: The present invention utilizes liposomes as a drug delivery system to effectively encapsulate hydrophobic celecoxib and saururusin, thereby improving their poor water solubility, thereby improving oral bioavailability and enabling the drugs to better exert their therapeutic effects; (2) High drug loading and encapsulation efficiency: The present invention adopts a specific formulation and preparation method, and the liposomes containing the active ingredients celecoxib and saururusin in Examples 1 to 3 have higher drug loading and encapsulation efficiency, wherein the addition of the polyvinyl alcohol-sodium alginate blend and the hydration temperature in the preparation method have a significant positive effect on the quality of the liposomes; (3) Synergistic therapeutic effect: The present invention uses saururaceous ketone and celecoxib in combination, and the two have a synergistic effect, which can more effectively reduce the arthritis index score of rats and alleviate the symptoms of rheumatoid arthritis. It is more effective in treating rheumatoid arthritis than using celecoxib alone. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 : Example 1 to Example 3, Comparative Example 1, Comparative Example 3, Comparative Example 4 The drug loading and encapsulation efficiency of celecoxib in liposomes containing the active ingredients celecoxib and saururusin.

[0019] Figure 2 : Example 1 to Example 3, Comparative Example 2 to Comparative Example 4 The drug loading and encapsulation efficiency of sauropodazone in liposomes containing the active ingredients celecoxib and sauropodazone.

[0020] Figure 3 : Arthritis index scores of rats in each group.

[0021] Figure 4 : The levels of tumor necrosis factor-α (TNF-α) in serum of rats in each group.

[0022] Figure 5 : Interleukin-6 (IL-6) content in serum of rats in each group. DETAILED DESCRIPTION

[0023] In order to make the purpose and technical solution of the present invention clearer, the present invention is further described below in conjunction with embodiments, but the protection scope of the present invention is not limited to these embodiments, and the embodiments are only used to explain the present invention. It should be understood by those skilled in the art that any changes or equivalent substitutions that do not deviate from the concept of the present invention are included in the protection scope of the present invention.

[0024] Example 1 Table 1. Example 1 Formulation: .

[0025] Preparation method: (1) Dissolve celecoxib and saururus in 100 ml of chloroform and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) dissolving the polyvinyl alcohol-sodium alginate blend in 100 ml of pH 6.0 buffer solution and preheating at 45°C, and adding it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and trifloxystrobin.

[0026] Example 2 Table 2. Example 2 Formulation: .

[0027] Preparation method: (1) Dissolve celecoxib and saururus in 200 ml of chloroform and ethanol (volume ratio 1:1) and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) dissolving the polyvinyl alcohol-sodium alginate blend in 200 ml of pH 5.5 buffer solution preheated at 40°C, and adding it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 200 bar for 5 cycles to obtain liposomes containing the active ingredients celecoxib and trifloxystrobin.

[0028] Example 3 Table 3. Example 3 Formulation: .

[0029] Preparation method: (1) Dissolve celecoxib and saururus in 150 ml of ethanol and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) dissolving the polyvinyl alcohol-sodium alginate blend in 150 ml of a buffer solution of pH 6.5 and preheating at 50°C, and adding the mixture to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 800 bar for two cycles to obtain liposomes containing the active ingredients celecoxib and trichloroacetic acid.

[0030] Comparative Example 1 Table 4. Comparative Example 1 Formula: .

[0031] Preparation method: (1) Dissolve celecoxib in 100 ml of chloroform and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) dissolving the polyvinyl alcohol-sodium alginate blend in 100 ml of pH 6.0 buffer solution and preheating at 45°C, and adding it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and trifloxystrobin.

[0032] Comparative Example 2 Table 5. Comparative Example 2 Formula: .

[0033] Preparation method: (1) Dissolve Saururaceae in 100 ml of chloroform and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) dissolving the polyvinyl alcohol-sodium alginate blend in 100 ml of pH 6.0 buffer solution and preheating at 45°C, and adding it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and trifloxystrobin.

[0034] Comparative Example 3 Table 6. Comparative Example 3 Formula: .

[0035] Preparation method: (1) Dissolve celecoxib and saururus in 100 ml of chloroform and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) Preheat 100 ml of pH 6.0 buffer solution at 45°C and add it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and trifloxystrobin.

[0036] Comparative Example 4 Table 7. Comparative Example 4 Formula: .

[0037] Preparation method: (1) Dissolve celecoxib and saururus in 100 ml of chloroform and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) Dissolve the polyvinyl alcohol-sodium alginate blend in 100 ml of pH 6.0 buffer solution (room temperature 25°C) and add it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and trifloxystrobin.

[0038] Comparative Example 5 Table 8. Comparative Example 5 Formula: .

[0039] Preparation method: (1) Dissolve celecoxib and saururus in 100 ml of chloroform and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) Dissolve polyvinyl alcohol in 100 ml of pH 6.0 buffer solution and preheat at 45°C, then add it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and trifloxystrobin.

[0040] Comparative Example 6 Table 9. Comparative Example 6 Formula: .

[0041] Preparation method: (1) Dissolve celecoxib and saururus in 100 ml of chloroform and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) Sodium alginate was dissolved in 100 ml of pH 6.0 buffer solution and preheated at 45°C, and then added to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and trifloxystrobin.

[0042] Comparative Example 7 Table 10. Comparative Example 7 Formulation: .

[0043] Preparation method: (1) Dissolve celecoxib and saururus in 100 ml of chloroform and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) dissolving the polyvinyl alcohol-sodium alginate blend in 100 ml of pH 6.0 buffer solution and preheating at 45°C, and adding it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes were homogenized under high pressure at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and trifloxystrobin.

[0044] Quality evaluation of liposomes containing active ingredients celecoxib and saururusin The contents of celecoxib and saururacetam in drug-loaded liposomes were detected by ultrafiltration and high performance liquid chromatography, and the drug loading and encapsulation efficiency of liposomes containing active ingredients celecoxib and saururacetam were calculated.

[0045] Table 11 Drug loading and encapsulation efficiency of liposomes in Comparative Examples 5 to 7 .

[0046] Figure 1 The drug loading amount and encapsulation efficiency of celecoxib in the liposomes containing the active ingredients celecoxib and saururusin in Example 1 to Example 3, Comparative Example 1, Comparative Example 3, and Comparative Example 4 are shown. Figure 2The drug loading amount and encapsulation efficiency of sauropodazone in the liposomes containing the active ingredients celecoxib and sauropodazone in Example 1 to Example 3 and Comparative Example 2 to Comparative Example 4 are shown. Figure 1 , Figure 2 Table 11 proves that the liposomes containing active ingredients celecoxib and saururus in Examples 1 to 3 of the present invention have higher drug loading and encapsulation efficiency. Experiments show that the addition of polyvinyl alcohol-sodium alginate blend and the temperature of hydration in the preparation method have a great influence on the quality of the liposomes. It should be noted that in the experiment to verify the effect of the ratio of polyvinyl alcohol-sodium alginate blend on the quality of liposomes, the ratio of polyvinyl alcohol-sodium alginate blend is 0.5:1, 2:1, 3:1, 4:1, 5:1, 6:1, 1:0.5, 2:1, 3:1, 4:1, 5:1, 6:1, and the effect is not as good as 1:1.

[0047] Pharmacodynamics experiments: SD rats, SPF grade, 180-220 g, experimental animal license number: SYXK (Lu) 20180008, were adaptively fed under standard conditions for 1 week before the experiment.

[0048] 1. Modeling: An emulsion of II collagen and Freund's incomplete adjuvant was injected at 2-3 cm from the base of the rat's tail to establish a rat model of rheumatoid arthritis. The rats with successful modeling were divided into a model group, Example 1 group, Example 2 group, Example 3 group, Comparative Example 1 group, and Comparative Example 2 group, with 10 rats in each group. Ten rats in the blank group were treated with sham surgery.

[0049] 2. Administration: Calculated with celecoxib, the dosage of Example 1 to Example 3 and Comparative Example 1 was 0.018 g / kg; the dosage of Comparative Example 2 was the same as that of the Example 1 group containing sauropodazone, and the dosage was administered for 7 days.

[0050] 3. Rat arthritis index score: The arthritis score of each rat was recorded according to the following scoring criteria: 0 points for no redness or swelling; 1 point for redness but no swelling of the toe joints; 2 points for mild redness and swelling of the toe joints; 3 points for moderate redness and swelling of the toe joints; 4 points for severe redness and swelling of the toe joints with impaired mobility. During each observation, the degree of swelling and mobility of the four paws of the rat should be noted and the score should be recorded accordingly.

[0051] Figure 3The arthritis index score of each group of rats was scored. The experimental results showed that there was no significant difference between the comparative example 2 group and the model group, proving that saururus trifoliatum was ineffective in treating rheumatoid arthritis; compared with the comparative example 1 group, the arthritis scores of rats in the example 1 group to the example 3 group were significantly different, P < 0.01, indicating that when saururus trifoliatum was used in combination with celecoxib, there was a synergistic effect between the two, which could more effectively reduce the arthritis index score of rats and alleviate the symptoms of rheumatoid arthritis, thereby showing a better effect in treating rheumatoid arthritis than using celecoxib alone.

[0052] Hematological index detection: Blood was collected from rats after anesthesia. The blood was centrifuged at 3000 rpm for 10 min and the supernatant was collected. ELISA was used to detect tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the serum.

[0053] Figure 4 is the content of tumor necrosis factor-α (TNF-α) in serum of rats in each group. Figure 5 is the interleukin-6 (IL-6) content in the serum of each group of rats. Compared with the comparative example 1 group, the TNF-α and IL-6 content in the serum of the rats in the example 1 group to the example 3 group had significant differences, P < 0.01, and the TNF-α and IL-6 content in the serum of the rats in the example 1 group to the example 3 group had significant differences compared with the model group, P < 0.01.

[0054] The pharmaceutical composition of the present invention can significantly reduce the arthritis score of rats with rheumatoid arthritis, significantly reduce the levels of TNF-a and IL-6 in the serum of rats, and improve the joint inflammation reaction of the body. The liposome containing the active ingredients celecoxib and saururusin is significantly better than celecoxib liposome in treating rheumatoid arthritis in rats, and has achieved remarkable progress.

[0055] The pharmacodynamic data of the present invention were statistically analyzed using SPSS 22.0 software, and the quantitative data were ( ). One-way analysis of variance was used for comparison among multiple groups, and independent sample T test was used for comparison between two groups. P < 0.05 was considered statistically significant.

Claims

1. A liposome containing the active ingredient celecoxib, characterized in that, The liposome is composed of 1 to 5 parts by weight of celecoxib, 0.3 to 0.8 parts by weight of sauropodazone, 4 to 8 parts by weight of phospholipids, 2 to 4 parts by weight of cholesterol, and 0.2 to 0.6 parts by weight of a polyvinyl alcohol-sodium alginate blend. The celecoxib structural formula is as follows: The structural formula of the saururaceous ketone is as follows: ; The mass ratio of polyvinyl alcohol to sodium alginate is 1:1; The preparation method of the liposome containing the active ingredients celecoxib and trichloroacetyl pyruvate is: (1) Dissolve celecoxib and saururus in an organic solvent and evaporate in a water bath at 30°C to 45°C until a lipid film appears; (2) dissolving the polyvinyl alcohol-sodium alginate blend in a buffer solution and preheating it at 40°C to 50°C, and adding it to the lipid film for hydration to form crude liposomes; (3) The crude liposomes are homogenized under high pressure to obtain liposomes containing the active ingredients celecoxib and trichloroacetic acid.

2. The liposome containing the active ingredient celecoxib according to claim 1, characterized in that: The phospholipids are egg yolk phospholipids and / or hydrogenated soybean phospholipids.

3. The liposome containing the active ingredient celecoxib according to claim 1, characterized in that: The organic solvent is chloroform and / or ethanol.

4. The liposome containing the active ingredient celecoxib according to claim 1, characterized in that: The buffer solution is selected from one of citrate buffer, phosphate buffer and acetate buffer.

5. The liposome containing the active ingredient celecoxib according to claim 1, characterized in that: The pH of the buffer solution is 5.5-6.

5.

6. The liposome containing the active ingredient celecoxib according to claim 1, characterized in that: The pressure of the high-pressure emulsification is 200-800 bar, and the cycle is 2-5 times.

7. An oral preparation, characterized in that The preparation consists of the liposome containing the active ingredient celecoxib as claimed in claim 1 and pharmaceutically acceptable excipients.

8. The oral preparation according to claim 7, characterized in that The oral preparations are tablets, granules, capsules and oral liquids.

9. The oral preparation according to claim 7, characterized in that The auxiliary materials are diluents, wetting agents, adhesives, disintegrants, lubricants, solubilizers, flavoring agents, preservatives, and colorants.

10. Use of the oral preparation according to claim 7 in preparing a drug for treating arthritis.

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