A liposome containing the active ingredient celecoxib, a preparation, and uses thereof

Abstract

The present invention belongs to the technical field of specific therapeutic activities of pharmaceutical preparations, and relates to a liposome containing the active ingredient celecoxib, a preparation, and uses thereof. The liposome of the present invention is prepared by the thin film dispersion method and is composed of celecoxib, sauchinone, phospholipid, cholesterol, and a polyvinyl alcohol-sodium alginate blend; the present invention uses the liposome to encapsulate celecoxib and sauchinone, improving their water solubility, and through a specific formula and method, the drug loading amount and encapsulation efficiency of the liposome are higher; in addition, there is a synergistic effect in the combined use of celecoxib and sauchinone, and the therapeutic effect on rheumatoid arthritis is better than that of using celecoxib alone.

Description

Technical Field

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

[0002] Disclosing the information of this background art section is only intended to enhance the understanding of the overall background of the present invention, and is not necessarily to be regarded as an admission or any form of implication that this information constitutes the prior art already known to those of ordinary skill in the art.

[0003] As a non-steroidal anti-inflammatory drug (NSAIDs), celecoxib is mainly used clinically to relieve the symptoms and signs of osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, and to treat acute pain in adults, etc. It selectively inhibits the activity of cyclooxygenase-2 (COX-2), reduces the synthesis of prostaglandins, thereby achieving anti-inflammatory, analgesic, and antipyretic effects. Compared with traditional non-steroidal anti-inflammatory drugs, celecoxib has relatively fewer adverse reactions on the gastrointestinal tract. However, celecoxib still has some limitations in clinical applications. First, celecoxib has poor water solubility and is almost insoluble in water, which results in low oral bioavailability. The dissolution of the drug in the gastrointestinal tract is a prerequisite for absorption. Due to the poor solubility of celecoxib, its dissolution rate in the gastrointestinal tract is slow, and the absorption is incomplete, affecting the efficacy of the drug. Second, the half-life of celecoxib is relatively short. In order to maintain an effective blood drug concentration, frequent administration is required, which not only brings inconvenience to patients, but also may lead to large fluctuations in blood drug concentration, increasing the risk of adverse reactions, such as adverse reactions in the cardiovascular system, etc.

[0004] Saururone (CAS: 177931-17-8) is an anti-inflammatory and antioxidant compound derived from natural plants. It belongs to anthocyanin compounds and has multiple biological activities such as anti-tumor, anti-inflammatory, and antioxidant. Research shows that saururone can regulate the immune system, reduce inflammatory responses, and to a certain extent inhibit the proliferation of tumor cells, having potential clinical application value. However, saururone has poor water solubility, resulting in low oral bioavailability and difficulty in fully exerting its efficacy.

[0005] To solve the above problems, liposomes, as an advanced drug delivery system, have gradually become an important tool for improving drug solubility, increasing drug bioavailability, and prolonging the drug action time. Liposomes are tiny lipid spheres composed of phospholipid molecules, which can effectively encapsulate hydrophobic and hydrophilic drugs, thereby improving the stability and bioavailability of the drugs. In addition, liposomes have the advantages of good biocompatibility, easy preparation, high drug loading capacity, etc., and can achieve slow release or targeted delivery of drugs during the drug release process, enhancing the therapeutic effect of the drugs. However, the research on liposome preparations containing both celecoxib and saururone is relatively less. Summary of the Invention

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

[0007] The structural formula of the said celecoxib is as follows: 。

[0008] The structural formula of the said sauchinone is as follows: 。

[0009] The said polyvinyl alcohol-sodium alginate blend is formed by mixing polyvinyl alcohol and sodium alginate according to a mass ratio of 1:1.

[0010] Further, the said phospholipid is egg yolk phospholipid or / and hydrogenated soy phospholipid.

[0011] The present invention also provides a preparation method of the liposome containing the active ingredients celecoxib and sauchinone, which belongs to the thin film dispersion method and is as follows:

[0012] (1) Dissolve celecoxib and sauchinone in an organic solvent, and rotary evaporate in a water bath at 30°C to 45°C until a lipid film appears;

[0013] (2) Dissolve the polyvinyl alcohol-sodium alginate blend in a buffer solution, preheat and keep warm at 40°C to 50°C, and add it to the lipid film for hydration to form a crude liposome;

[0014] (3) Subject the crude liposome to high-pressure homogenization to obtain the liposome containing the active ingredients celecoxib and sauchinone.

[0015] Further, the said organic solvent is chloroform or / and ethanol; the said buffer solution is selected from one of citrate buffer solution, phosphate buffer solution, and acetate buffer solution; the pH of the said buffer solution is 5.5 to 6.5; the pressure of the said high-pressure homogenization is 200 to 800 bar, and it is circulated 2 to 5 times.

[0016] The present invention also provides an oral preparation, which is composed of the above-mentioned liposome containing the active ingredients celecoxib and sauchinone and pharmaceutically acceptable excipients. The oral preparation is tablets, granules, capsules, or oral liquids. The excipients are diluents, wetting agents, binders, disintegrants, lubricants, solubilizers, flavoring agents, preservatives, and colorants.

[0017] The preparation of the tablets is as follows: Take the above-mentioned liposomes containing the active ingredients celecoxib and sauchinone, freeze-dry them into a powder, add one or more of starch, mannitol, dextrin, lactose, and microcrystalline cellulose, add one or more of sodium carboxymethyl starch (CMS-Na), crospovidone (PVPP), croscarmellose sodium (CCMC-Na), and low-substituted hydroxypropyl cellulose, add one or more of magnesium stearate, colloidal silicon dioxide, talc, and polyethylene glycol, and then press into tablets. The preparation of the tablets can also be carried out by wet granulation: Take the above-mentioned liposomes containing the active ingredients celecoxib and sauchinone, concentrate them into a paste with a relative density of 1.4 to 1.6, add one or more of starch, mannitol, dextrin, lactose, and microcrystalline cellulose, add one or more of sodium carboxymethyl starch (CMS-Na), crospovidone (PVPP), croscarmellose sodium (CCMC-Na), and low-substituted hydroxypropyl cellulose, carry out wet granulation, screen the granules, add one or more of magnesium stearate, colloidal silicon dioxide, talc, and polyethylene glycol, and then press into tablets.

[0018] The preparation of the oral liquid is as follows: Take the above-mentioned liposomes containing the active ingredients celecoxib and sauchinone, add one or more of ethanol, propylene glycol, polysorbate 80, and poloxamer, add one or more of sucrose, stevioside, aspartame, and essence, add one or more of parabens (p-hydroxybenzoates), benzoic acid and its sodium salt, sorbic acid and its potassium salt, and benzalkonium bromide, and mix them to obtain the oral liquid.

[0019] The present invention also provides the use of the above-mentioned oral preparations containing the active ingredients celecoxib and sauchinone in the preparation of drugs for treating arthritis.

[0020] Compared with the prior art, the technical effects of the present invention are as follows:

[0021] (1) Improving drug solubility and bioavailability: The present invention utilizes the drug delivery system of liposomes to effectively encapsulate the hydrophobic celecoxib and sauchinone, improving the problem of their poor water solubility, thereby enhancing the oral bioavailability and enabling the drug to exert its efficacy better;

[0022] (2) High drug loading and encapsulation efficiency: Through specific formulations and preparation methods, the liposomes containing the active ingredients celecoxib and sauchinone in Examples 1 to 3 have higher drug loading and encapsulation efficiency. The addition of the polyvinyl alcohol-sodium alginate blend and the hydration temperature in the preparation method have a greater positive impact on the quality of the liposomes;

[0023] (3) Synergistic treatment effect: In the present invention, the combined use of sauchinone and celecoxib has a synergistic effect, which can more effectively reduce the arthritis index score of rats and alleviate the symptoms of rheumatoid arthritis, and has a better effect in treating rheumatoid arthritis than the single use of celecoxib. Description of the Drawings

[0024] Figure 1 : Drug loading and encapsulation efficiency of celecoxib in liposomes containing active ingredients celecoxib and sauchinone in Examples 1 to 3, Comparative Example 1, Comparative Example 3, and Comparative Example 4.

[0025] Figure 2 : Drug loading and encapsulation efficiency of sauchinone in liposomes containing active ingredients celecoxib and sauchinone in Examples 1 to 3, Comparative Example 2 to Comparative Example 4.

[0026] Figure 3 : Arthritis index scores of rats in each group.

[0027] Figure 4 : Content of tumor necrosis factor-α (TNF-α) in the serum of rats in each group.

[0028] Figure 5 : Content of interleukin-6 (IL-6) in the serum of rats in each group. Detailed Description of the Invention

[0029] In order to make the objectives and technical solutions of the present invention clearer, the following further describes the present invention with reference to the embodiments. However, the protection scope of the present invention is not limited to these embodiments, and the embodiments are only used to explain the present invention. Those skilled in the art should understand 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.

[0030] Example 1

[0031] Table 1. Formulation of Example 1:

[0032] 。

[0033] Preparation method:

[0034] (1) Dissolve celecoxib and sauchinone in 100 ml of chloroform, and rotary evaporate in a water bath at 30°C to 45°C until a lipid film appears;

[0035] (2) Dissolve the polyvinyl alcohol-sodium alginate blend in 100 ml of a buffer solution with a pH of 6.0 and preheat it at 45°C, then add it to the lipid film for hydration to form crude liposomes;

[0036] (3) Subject the crude liposomes to high-pressure homogenization at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0037] Example 2

[0038] Table 2. Formulation of Example 2:

[0039] .

[0040] Preparation method:

[0041] (1) Dissolve celecoxib and sauchinone in 200 ml of chloroform and ethanol (volume ratio 1:1), and rotary evaporate in a water bath at 30°C to 45°C until a lipid film appears;

[0042] (2) Dissolve the polyvinyl alcohol-sodium alginate blend in 200 ml of a buffer solution with pH = 5.5, preheat at 40°C, and add it to the lipid film for hydration to form crude liposomes;

[0043] (3) Subject the crude liposomes to high-pressure homogenization at a pressure of 200 bar for 5 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0044] Example 3

[0045] Table 3. Formulation of Example 3:

[0046] .

[0047] Preparation method:

[0048] (1) Dissolve celecoxib and sauchinone in 150 ml of ethanol, and rotary evaporate in a water bath at 30°C to 45°C until a lipid film appears;

[0049] (2) Dissolve the polyvinyl alcohol-sodium alginate blend in 150 ml of a buffer solution with pH = 6.5, preheat at 50°C, and add it to the lipid film for hydration to form crude liposomes;

[0050] (3) Subject the crude liposomes to high-pressure homogenization at a pressure of 800 bar for 2 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0051] Comparative Example 1

[0052] Table 4. Formulation of Comparative Example 1:

[0053] .

[0054] Preparation method:

[0055] (1) Dissolve celecoxib in 100 ml of chloroform and rotary evaporate it in a water bath at 30°C - 45°C until a lipid film appears;

[0056] (2) Dissolve the polyvinyl alcohol - sodium alginate blend in 100 ml of buffer solution with pH = 6.0, preheat it at 45°C, add it to the lipid film for hydration to form crude liposomes;

[0057] (3) Subject the crude liposomes to high - pressure homogenization at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0058] Comparative Example 2

[0059] Table 5. Formulation of Comparative Example 2:

[0060] 。

[0061] Preparation method:

[0062] (1) Dissolve sauchinone in 100 ml of chloroform and rotary evaporate it in a water bath at 30°C - 45°C until a lipid film appears;

[0063] (2) Dissolve the polyvinyl alcohol - sodium alginate blend in 100 ml of buffer solution with pH = 6.0, preheat it at 45°C, add it to the lipid film for hydration to form crude liposomes;

[0064] (3) Subject the crude liposomes to high - pressure homogenization at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0065] Comparative Example 3

[0066] Table 6. Formulation of Comparative Example 3:

[0067] 。

[0068] Preparation method:

[0069] (1) Dissolve celecoxib and sauchinone in 100 ml of chloroform and rotary evaporate it in a water bath at 30°C - 45°C until a lipid film appears;

[0070] (2) Preheat 100 ml of buffer solution with pH = 6.0 at 45°C, add it to the lipid film for hydration to form crude liposomes;

[0071] (3) Subject the crude liposomes to high - pressure homogenization at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0072] Comparative Example 4

[0073] Table 7. Formulation of Comparative Example 4:

[0074] 。

[0075] Preparation method:

[0076] (1) Dissolve celecoxib and sauchinone in 100 ml of chloroform, and rotary evaporate in a water bath at 30°C - 45°C until a lipid film appears;

[0077] (2) Dissolve the polyvinyl alcohol - sodium alginate blend in 100 ml of buffer solution with pH = 6.0 (at room temperature of 25°C), add it to the lipid film for hydration to form crude liposomes;

[0078] (3) Subject the crude liposomes to high - pressure homogenization at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0079] Comparative Example 5

[0080] Table 8. Formulation of Comparative Example 5:

[0081] 。

[0082] Preparation method:

[0083] (1) Dissolve celecoxib and sauchinone in 100 ml of chloroform, and rotary evaporate in a water bath at 30°C - 45°C until a lipid film appears;

[0084] (2) Dissolve polyvinyl alcohol in 100 ml of buffer solution with pH = 6.0 and pre - heat at 45°C, add it to the lipid film for hydration to form crude liposomes;

[0085] (3) Subject the crude liposomes to high - pressure homogenization at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0086] Comparative Example 6

[0087] Table 9. Formulation of Comparative Example 6:

[0088] 。

[0089] Preparation method:

[0090] (1) Dissolve celecoxib and sauchinone in 100 ml of chloroform, and rotary evaporate in a water bath at 30°C - 45°C until a lipid film appears;

[0091] (2) Dissolve sodium alginate in 100 ml of buffer solution with pH = 6.0 and pre - heat at 45°C, add it to the lipid film for hydration to form crude liposomes;

[0092] (3) High-pressure homogenize the crude liposomes at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0093] Comparative Example 7

[0094] Table 10. Formulation of Comparative Example 7:

[0095] .

[0096] Preparation method:

[0097] (1) Dissolve celecoxib and sauchinone in 100 ml of chloroform and rotary evaporate in a water bath at 30°C to 45°C until a lipid film appears;

[0098] (2) Dissolve the polyvinyl alcohol-sodium alginate blend in 100 ml of buffer solution with pH = 6.0 and preheat at 45°C, then add it to the lipid film for hydration to form crude liposomes;

[0099] (3) High-pressure homogenize the crude liposomes at a pressure of 500 bar for 3 cycles to obtain liposomes containing the active ingredients celecoxib and sauchinone.

[0100] Quality evaluation of liposomes containing the active ingredients celecoxib and sauchinone

[0101] Use the ultrafiltration method and high-performance liquid chromatography to detect the contents of celecoxib and sauchinone in the drug-loaded liposomes, and calculate the drug loading and encapsulation efficiency of the liposomes containing the active ingredients celecoxib and sauchinone.

[0102] Table 11 Drug loading and encapsulation efficiency of liposomes in Comparative Examples 5 to 7

[0103] .

[0104] Figure 1 For the drug loading and encapsulation efficiency of celecoxib in the liposomes containing the active ingredients celecoxib and sauchinone in Examples 1 to 3, Comparative Example 1, Comparative Example 3, and Comparative Example 4. Figure 2 For the drug loading and encapsulation efficiency of sauchinone in the liposomes containing the active ingredients celecoxib and sauchinone in Examples 1 to 3, Comparative Example 2 to Comparative Example 4. Figure 1 、 Figure 2Table 11 demonstrates that the liposomes containing the active ingredients celecoxib and sauchinone in Examples 1 to 3 of the present invention have higher drug loading and encapsulation efficiency. Experiments have shown that the addition of the polyvinyl alcohol-sodium alginate blend and the hydration temperature in the preparation method have a greater impact on the quality of the liposomes. It should be noted that in the experiment to verify the effect of the polyvinyl alcohol-sodium alginate blend ratio on the quality of the liposomes, the effects of the polyvinyl alcohol-sodium alginate blend ratios of 0.5:1, 2:1, 3:1, 4:1, 5:1, 6:1, 1:0.5, 2:1, 3:1, 4:1, 5:1, and 6:1 are all inferior to 1:1.

[0105] Pharmacodynamic experiment:

[0106] SD rats, SPF grade, 180 - 220 g, experimental animal license number: SYXK(Shandong) 20180008, were adaptively fed under standard conditions for 1 week before the experiment.

[0107] 1. Model establishment: At 2 - 3 cm from the root of the rat tail, an emulsion of type II collagen and Freund's incomplete adjuvant was injected to establish a rat model of rheumatoid arthritis. The successfully modeled rats were divided into a model group, an Example 1 group, an Example 2 group, an Example 3 group, a Comparative Example 1 group, and a Comparative Example 2 group, with 10 rats in each group. 10 rats in the blank group were given sham operation.

[0108] 2. Administration: Calculated by celecoxib, the dose for the Example 1 group to the Example 3 group and the Comparative Example 1 group was 0.018 g / kg; the administration dose of the Comparative Example 2 group was the same as the dose of sauchinone contained in the administration of the Example 1 group, and the administration was for 7 days.

[0109] 3. Arthritis index score for rats:

[0110] Record the arthritis score of each rat according to the following scoring criteria: 0 points indicates no redness or swelling; 1 point indicates redness of the toe joint without swelling; 2 points indicates mild redness and swelling of the toe joint; 3 points indicates moderate redness and swelling of the toe joint; 4 points indicates severe redness and swelling of the toe joint with movement disorder. Each time of observation, pay attention to the swelling degree and mobility of the 4 paw pads of the rat and record the score accordingly.

[0111] Figure 3 The arthritis index scores for each group of rats are as follows. The experimental results show that there is no significant difference between the Comparative Example 2 group and the model group, proving that sauchinone is ineffective in the treatment of rheumatoid arthritis; compared with the Comparative Example 1 group, there is a significant difference in the arthritis scores of the rats in the Example 1 group to the Example 3 group, P < 0.01, indicating that when sauchinone and celecoxib are used in combination, there is a synergistic effect between the two, which can more effectively reduce the arthritis index score of rats and relieve the symptoms of rheumatoid arthritis, thus showing a better effect in the treatment of rheumatoid arthritis than using celecoxib alone.

[0112] Hematological index detection:

[0113] After the rats were anesthetized, blood was collected. After the blood collection, the supernatant was collected after centrifugation at 3000 rpm at low temperature for 10 min. The levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the serum were detected by ELISA method.

[0114] Figure 4 It was the content of tumor necrosis factor-α (TNF-α) in the serum of rats in each group. Figure 5 It was the content of interleukin-6 (IL-6) in the serum of rats in each group. Compared with the control group 1, there were significant differences in the contents of TNF-α and IL-6 in the serum of rats in the experimental groups 1 to 3, P < 0.01. Compared with the model group, there were significant differences in the contents of TNF-α and IL-6 in the serum of rats in the experimental groups 1 to 3, P < 0.01.

[0115] The pharmaceutical composition of the present invention can significantly reduce the arthritis score of rats with rheumatoid arthritis, significantly reduce the levels of TNF-α and IL-6 in the serum of rats, and improve the joint inflammatory reaction of the body. The effect of the liposome containing the active ingredients celecoxib and sauchinone in treating rheumatoid arthritis in rats is significantly better than that of celecoxib liposome, and significant progress has been made.

[0116] The pharmacodynamic data of the present invention were statistically analyzed using SPSS 22.0 software, and the measurement data were expressed as ( ). One-way ANOVA was used for comparison among multiple groups, and independent sample T-test was used between two groups. P < 0.05 was considered as having statistical significance.

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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