Preparation method and application of nepafenac and quercetin sustained-release gel preparation

Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a preparation method and application of a nepafenac and quercetin sustained-release gel preparation, and the nepafenac and quercetin sustained-release gel preparation comprises the following components in parts by mass: 5-75 parts of nepafenac; 3 to 60 parts of quercetin; 100 to 500 parts of a biological carrier; 15 to 350 parts of a viscosity reducing agent; and 50 to 200 parts of a non-aqueous solvent. The composition can relieve pain and resist inflammation, can promote tissue repair and growth, can effectively realize a slow release effect, and is convenient and long-acting in administration. The invention also provides a preparation method and application thereof.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a preparation method and application of a nepafenac and quercetin sustained-release gel preparation. Background Art

[0002] Diabetic foot (DF) refers to foot infections, ulcers, and / or deep tissue damage associated with local nerve abnormalities and peripheral vascular disease in the distal lower limbs in patients with diabetes. According to the "Guidelines for the Diagnosis and Treatment of Diabetic Foot in China (2023)", the prevalence of foot ulcers in diabetic patients is 4%-10%, of which 70% require surgical treatment, and the amputation rate of diabetic foot is 14%-22%, with a high disability and mortality rate. Diabetic foot is difficult to cure and has a poor prognosis. It requires multidisciplinary collaborative comprehensive treatment involving wound repair, vascular surgery, orthopedics, endocrinology, rehabilitation and other disciplines. At present, there are three main clinical treatment methods: maintenance, infection prevention, and nerve nutrition, but none of them can fundamentally regenerate tissues and promote wound healing.

[0003] Nepafenac is a new type of nonsteroidal antipyretic, analgesic and anti-inflammatory drug (NSAIDs), which can be converted into aminofenac under the action of tissue hydrolases. Amfenac can inhibit COX-1 and COX-2 to inhibit angiogenesis and significantly reduce prostaglandin levels. Compared with traditional nonsteroidal antipyretic, analgesic and anti-inflammatory drugs, nepafenac has the advantages of strong penetration, strong targeting effect, and small toxic side effects. Due to its low solubility and low bioavailability, but its ability to quickly reach the target location and exert its effects, nepafenac is currently mostly used in the form of suspension in clinical practice to reduce pain and inflammation in diabetic patients after cataract surgery and reduce the risk of macular edema in diabetic patients after surgery. Studies have shown that nepafenac is also a promising drug in the treatment of skin diseases and skin ulcers with inflammatory immune-allergic components.

[0004] Quercetin, an active ingredient in traditional Chinese medicine, has the function of regulating immune cells. It inhibits the expression of inflammatory factors by balancing pro-inflammatory and anti-inflammatory cytokines, reduces the inflammatory response induced by high sugar, and plays a protective role in peripheral neuropathy. It protects cells around the wound from damage by scavenging free radicals and inhibiting oxidative stress, promotes tissue repair, and reduces scar tissue formation. It can also improve the microcirculation in the diabetic foot wound area, increase blood supply, provide sufficient oxygen and nutrients, promote the proliferation and migration of fibroblasts and keratinocytes, and accelerate wound healing.

[0005] Poly(orthoesters) are a class of biodegradable polymer materials with potential applications in the field of biology. Poly(orthoesters) have surface hydrophobicity and can be dissolved in certain organic solvents, such as tetrahydrofuran, dichloromethane and cyclohexane. The drug release kinetics and polymer degradation rate of poly(orthoesters) follow zero-order kinetics without any sudden effects. The good sustained-release effect and high safety characteristics exhibited by poly(orthoesters) make them potentially useful in drug delivery systems, especially in the development of sustained-release or controlled-release drug preparations.

[0006] The chemical structure of polyorthoester is: .

[0007] Although the treatment goals of diabetic foot are clear, namely to relieve pain, control inflammation and prevent infection, the existing therapies have significant shortcomings. The efficacy is short-lived and the medication needs to be changed frequently, which affects patient compliance. In addition, the inconvenience of treatment also makes it difficult for patients to adhere to it. Therefore, the treatment scheme of convenient and more effective new drug delivery preparations shows broad application prospects in the field of diabetic foot treatment. Summary of the invention

[0008] In order to solve the above technical problems, the present invention provides a nepafenac and quercetin sustained-release gel preparation, which not only relieves pain and inflammation but also promotes tissue repair and growth, can achieve a sustained-release effect, and is convenient and long-lasting in administration. The present invention also provides a preparation method and application thereof.

[0009] The nepafenac and quercetin sustained-release gel preparation of the present invention comprises the following components in parts by weight: Nepafenac, 5-75 parts; Quercetin, 3-60 parts; Biological carrier, 100-500 copies; Viscosity reducer, 15-350 parts; Non-aqueous solvent, 50-200 parts.

[0010] Preferably, the biocarrier is a polyorthoester.

[0011] Preferably, the viscosity reducer is one of ethyl acetate and triacetin, more preferably triacetin.

[0012] Preferably, the non-aqueous solvent is one of dimethyl sulfoxide and N-methylpyrrolidone, more preferably dimethyl sulfoxide.

[0013] The preparation method of the nepafenac and quercetin sustained-release gel preparation of the present invention comprises the following steps: (1) Heat the biocarrier in an oil bath at 55-75°C for 1-2 hours to improve fluidity; (2) mixing the heated biological carrier in step (1) with the viscosity reducer, stirring magnetically for 0.5 h, and then adding the non-aqueous solvent, nepafenac and quercetin respectively, and continuing magnetic stirring at a temperature of 50-70° C. and a stirring speed of 100-300 rpm to obtain a uniform solution; (3) sterilizing and filtering the uniform solution obtained in step (2) through a filter under aseptic conditions at a filtration temperature of 25-37° C. and a filtration accuracy of 0.45 μm to obtain a drug solution; (4) Dispense the sterilized liquid into vials.

[0014] The application of the nepafenac and quercetin sustained-release gel preparation of the present invention in preparing a drug for treating diabetic foot. The drug can be administered topically.

[0015] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention uses polyorthoester biocarrier as a bioadhesive controlled-release skeleton material, not only because it has good biocompatibility, tissue adhesion and drug release blocking properties, but also can load small molecule drugs and macromolecular drugs. Polyorthoester has a high drug loading capacity, can effectively reduce the sudden release of drugs, reduce the side effects of drug degradation, cover the time of the peak of pain, and achieve a 3-5 day sustained-release effect. In addition, the final degradation product of polyorthoester in the body is a water-soluble small molecule, which is easily metabolized by the organism; 2. The preparation of the present invention can be directly injected or applied to the wound at the debridement site. Compared with other dosage forms, it has the advantages of convenient administration, forming a drug reservoir with stable morphology and sustainable release at the administration site, prolonging the action time and improving the therapeutic effect of combined medication, and also improving the patient's medication compliance and tolerance; 3. The two active ingredients nepafenac and quercetin contained in the present invention can not only relieve pain and inflammation but also promote tissue repair and growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is a graph of the in vitro cumulative release of nepafenac from the preparations of Examples 1-4 and Comparative Examples 1-4 at different times; Figure 2 It is the in vitro cumulative release curve of quercetin of the preparations of Examples 1-4 and Comparative Examples 1-4 at different times. DETAILED DESCRIPTION

[0017] The technical solution of the present invention will be clearly and completely described below in conjunction with the accompanying drawings and embodiments.

[0018] Unless otherwise specified, all raw materials used in the examples are commercially available.

[0019] Among them, nepafenac, Guangzhou Yanlord Pharmaceutical Technology Co., Ltd.; Quercetin, Nanjing Tianrun Biological Co., Ltd.; Polyorthoesters, Shandong Qidu Pharmaceutical Co., Ltd.

[0020] Example 1 The nepafenac and quercetin sustained-release gel preparation comprises the following components in parts by weight: Nepafenac, 50 parts; quercetin, 45 parts; Biocarrier polyorthoester, 350 parts; Viscosity reducing agent triacetin, 225 parts; Non-aqueous solvent dimethyl sulfoxide, 155 parts.

[0021] The preparation method comprises the following steps: (1) Heat the biocarrier polyorthoester in an oil bath at 55°C for 1 h; (2) mixing the heated polyorthoester in step (1) with triacetin, stirring magnetically for 0.5 h, and then adding dimethyl sulfoxide, nepafenac and quercetin respectively, and continuing magnetic stirring at a temperature of 55° C. and a stirring speed of 100 rpm to obtain a uniform solution; (3) sterilizing the uniform solution obtained in step (2) through a filter under aseptic conditions at a filtration temperature of 30° C. and a filtration accuracy of 0.45 μm to obtain a drug solution; (4) Dispense the sterilized liquid into vials.

[0022] Example 2 The nepafenac and quercetin sustained-release gel preparation comprises the following components in parts by weight: Nepafenac, 50 parts; quercetin, 45 parts; Biocarrier polyorthoester, 500 parts; Viscosity reducing agent triacetin, 350 parts; Non-aqueous solvent N-methylpyrrolidone, 200 parts.

[0023] The preparation method comprises the following steps: (1) Heat the biocarrier polyorthoester in an oil bath at 60°C for 2 h; (2) mixing the heated polyorthoester in step (1) with triacetin, stirring magnetically for 0.5 h, and then adding N-methylpyrrolidone, nepafenac and quercetin respectively, continuing magnetic stirring at a temperature of 60° C. and a stirring speed of 300 rpm to obtain a uniform solution; (3) sterilizing the uniform solution obtained in step (2) through a filter under aseptic conditions at a filtration temperature of 25° C. and a filtration accuracy of 0.45 μm to obtain a drug solution; (4) Dispense the sterilized liquid into vials.

[0024] Example 3 The nepafenac and quercetin sustained-release gel preparation comprises the following components in parts by weight: Nepafenac, 5 parts; quercetin, 3 parts; Biocarrier polyorthoester, 100 parts; Viscosity reducing agent ethyl acetate, 150 parts; Non-aqueous solvent dimethyl sulfoxide, 100 parts.

[0025] The preparation method comprises the following steps: (1) Heat the polyorthoester in an oil bath at 60°C for 2 h; (2) mixing the heated polyorthoester in step (1) with ethyl acetate, stirring magnetically for 0.5 h, and then adding dimethyl sulfoxide, nepafenac and quercetin respectively, and continuing magnetic stirring at a temperature of 60° C. and a stirring speed of 200 rpm to obtain a uniform solution; (3) filtering the uniform solution obtained in step (2) through a filter under sterile conditions at a temperature of 37° C. and a filtration accuracy of 0.45 μm to obtain a drug solution; (4) Dispense the sterilized liquid into vials.

[0026] Example 4 The nepafenac and quercetin sustained-release gel preparation comprises the following components in parts by weight: Nepafenac, 75 parts; quercetin, 60 parts; Biocarrier polyorthoester, 150 parts; Viscosity reducing agent triacetin, 15 parts; Non-aqueous solvent dimethyl sulfoxide, 50 parts.

[0027] The preparation method comprises the following steps: (1) Heat the polyorthoester in an oil bath at 70°C for 2 h; (2) mixing the heated polyorthoester in step (1) with triacetin, stirring magnetically for 0.5 h, and then adding dimethyl sulfoxide, nepafenac and quercetin respectively, and continuing magnetic stirring at a temperature of 60° C. and a stirring speed of 200 rpm to obtain a uniform solution; (3) sterilizing the uniform solution obtained in step (2) through a filter under aseptic conditions at a filtration temperature of 30° C. and a filtration accuracy of 0.45 μm to obtain a drug solution; (4) Dispense the sterilized liquid into vials.

[0028] Comparative Example 1 Nepafenac and quercetin sustained-release gel preparation, comprising the following components in parts by weight: Nepafenac, 50 parts; quercetin, 45 parts; Viscosity reducing agent triacetin, 225 parts; Non-aqueous solvent dimethyl sulfoxide, 155 parts.

[0029] The preparation method comprises the following steps: (1) Dimethyl sulfoxide, nepafenac and quercetin were added to triacetin respectively, and the mixture was stirred magnetically at a temperature of 55°C and a speed of 200 rpm to obtain a uniform solution; (2) filtering the uniform solution obtained in step (2) through a filter under sterile conditions at a temperature of 30° C. and a filtration accuracy of 0.45 μm to obtain a drug solution; (3) Dispense the sterilized liquid into vials.

[0030] Comparative Example 2 Nepafenac and quercetin sustained-release gel preparation, comprising the following components in parts by weight: Nepafenac, 50 parts; quercetin, 45 parts; Hyaluronic acid, 200 parts; Phosphate buffer solution, 380 parts.

[0031] The preparation method comprises the following steps: (1) mixing hyaluronic acid and phosphate buffer solution, stirring with a magnetic stirrer at a speed of 100 rpm and a temperature of 40° C. to obtain solution 1; (2) adding nepafenac and quercetin to solution 1, stirring with magnetic force at a temperature of 55°C and a stirring speed of 200 rpm to obtain a uniform solution; (3) sterilizing the uniform solution obtained in step (2) through a filter under aseptic conditions at a filtration temperature of 30° C. and a filtration accuracy of 0.45 μm to obtain a drug solution; (4) Dispense the sterilized liquid into vials.

[0032] Comparative Example 3 Nepafenac and quercetin sustained-release gel preparation, comprising the following components in parts by weight: Nepafenac, 50 parts; quercetin, 45 parts; Chitosan, 200 parts; Acetic acid solution, 380 parts.

[0033] The preparation method comprises the following steps: (1) mixing chitosan and acetic acid solution, stirring with magnetic force at a stirring speed of 100 rpm and a temperature of 40° C. to obtain a first solution; (2) adding nepafenac and quercetin to the first solution of step (1), continuing magnetic stirring at a temperature of 55° C. and a stirring speed of 200 rpm to obtain a uniform solution; (3) sterilizing the uniform solution obtained in step (2) through a filter under aseptic conditions at a filtration temperature of 30° C. and a filtration accuracy of 0.45 μm to obtain a drug solution; (4) Dispense the sterilized liquid into vials.

[0034] Comparative Example 4 Nepafenac and quercetin sustained-release gel preparation, comprising the following components in parts by weight: Nepafenac, 50 parts; quercetin, 45 parts; Biocarrier polyorthoester, 350 parts; Non-aqueous solvent dimethyl sulfoxide, 155 parts.

[0035] The preparation method comprises the following steps: (1) Heat the biocarrier polyorthoester in an oil bath at 55°C for 1 h; (2) adding dimethyl sulfoxide, nepafenac and quercetin to the polyorthoester heated in step (1), and continuing magnetic stirring at a temperature of 55° C. and a stirring speed of 200 rpm to obtain a uniform solution; (3) sterilizing the uniform solution obtained in step (2) through a filter under aseptic conditions at a filtration temperature of 30° C. and a filtration accuracy of 0.45 μm to obtain a drug solution; (4) Dispense the sterilized liquid into vials.

[0036] Performance Testing Investigation of in vitro release: Examples 1-4 and Comparative Examples 1-4 were subjected to release determination according to the release determination method. The sustained-release gel preparation containing 50 mg of nepafenac and 45 mg of quercetin per sample was placed in a dissolution cup, and 250 mL of a medium containing 0.025 M phosphate buffer (3.9 g / L NaH2PO4·2H2O, pH adjusted to 7.4±0.05 with sodium hydroxide test solution) was added. The medium was placed in a dissolution apparatus at 37°C±0.5°C at a speed of 100 rpm. 1.0 mL of samples were taken at 0h, 12h, 24h, 48h, 72h, 96h, and 120h without rehydration.

[0037] The sample was filtered through a 0.45 μm filter membrane as the test sample solution; according to the high performance liquid chromatography method (General Rules 0512 of the Fourth Part of the 2020 Edition of the Chinese Pharmacopoeia), the Shim-Pack-GIST C18 column was used, with specifications of 4.6×250 mm, 5 μm, and 0.02 mol / L phosphate solution and acetonitrile in a volume ratio of 80:20 as mobile phase A, 0.02 mol / L phosphate solution and acetonitrile in a volume ratio of 30:70 as mobile phase B, the flow rate was 1.0 mL / min, the column temperature was 35 ° C, the detection wavelength was 278 nm, and the injection volume was 20 μL. Separately, accurately weigh an appropriate amount of nepafenac and quercetin reference substances, dissolve and dilute with 80% acetonitrile solution to make a solution containing about 0.1 mg of nepafenac and quercetin per 1 mL as the reference solution. The cumulative release at different times was calculated by the external standard method using the peak area, and a cumulative release curve was plotted. The results of the cumulative release of nepafenac are shown in Table 1, and the cumulative release curve is shown in Table 1. Figure 1 The results of the cumulative release of quercetin are shown in Table 2, and the cumulative release curve is shown in Figure 2 shown.

[0038] Table 1 Determination results of cumulative release of nepafenac in Examples 1-4 and Comparative Examples 1-4

[0039] Table 2 Determination results of cumulative release of quercetin in Examples 1-4 and Comparative Examples 1-4

[0040] From Table 1, Table 2 and Figure 1 , Figure 2It can be concluded that the nepafenac and quercetin sustained-release gel preparations prepared in Examples 1-4 have an obvious sustained-release effect on the active ingredients nepafenac and quercetin, indicating that the polyorthoester polymer biological carrier in the system can improve the stability of the drug, slowly release the drug in the preparation, prolong the action time, thereby reducing the number of administrations and greatly improving the patient's medication compliance. In Comparative Example 1, the biological carrier polyorthoester is lacking, and the preparation is released in a short time, and the sustained-release effect is not achieved; in Comparative Examples 2 and 3, different water-soluble carrier matrices are used, and the sustained-release effect cannot be achieved in a short time, proving that the water-soluble carrier system is not suitable for the present invention. In Comparative Example 4, the viscosity reducer is lacking, and although the preparation can achieve sustained release, its unit release amount is significantly reduced, which is not conducive to the effect of the active ingredient. The biological carrier polyorthoester in the present invention is combined with a non-aqueous solvent and a viscosity reducer to form an effective sustained-release preparation system, which can effectively reduce the burst release and achieve a better sustained-release effect.

Claims

1. A nepafenac and quercetin sustained-release gel preparation, characterized in that: The composition includes the following components in parts by weight: Nepafenac, 5-75 parts; Quercetin, 3-60 parts; Biological carrier, 100-500 copies; Viscosity reducer, 15-350 parts; Non-aqueous solvent, 50-200 parts.

2. The nepafenac and quercetin sustained-release gel preparation according to claim 1, characterized in that: The biological carrier is a polyorthoester.

3. The nepafenac and quercetin sustained-release gel preparation according to claim 1, characterized in that: The viscosity reducing agent is one of ethyl acetate and triacetin.

4. The nepafenac and quercetin sustained-release gel preparation according to claim 1, characterized in that: The non-aqueous solvent is one of dimethyl sulfoxide and N-methylpyrrolidone.

5. A method for preparing the nepafenac and quercetin sustained-release gel preparation according to any one of claims 1 to 4, characterized in that: The following steps are involved: (1) Heat the biocarrier in an oil bath; (2) mixing the heated biological carrier in step (1) with the viscosity reducer, and then adding a non-aqueous solvent, nepafenac and quercetin respectively to obtain a uniform solution; (3) Aseptically filter the uniform solution obtained in step (2) to obtain a drug solution.

6. The method for preparing the nepafenac and quercetin sustained-release gel preparation according to claim 5, characterized in that: In step (1), the heating temperature is 55-75°C and the heating time is 1-2h.

7. The method for preparing the nepafenac and quercetin sustained-release gel preparation according to claim 5, characterized in that: Step (2) is specifically as follows: the heated biological carrier in step (1) is mixed with the viscosity reducer, and magnetically stirred for 0.5 h, and then the non-aqueous solvent, nepafenac and quercetin are added respectively, and the magnetic stirring is continued at a stirring temperature of 50-70° C. and a stirring speed of 100-300 rpm to obtain a uniform solution.

8. The method for preparing the nepafenac and quercetin sustained-release gel preparation according to claim 5, characterized in that: In step (3), the filtration temperature is 25-37°C and the filtration accuracy is 0.45 μm.

9. Use of the nepafenac and quercetin sustained-release gel preparation according to any one of claims 1 to 4 for preparing a drug for treating diabetic foot.

Images