Gel ointment for treating osteoarthritis and preparation method thereof

Abstract

The invention discloses a gel ointment for treating osteoarthritis and a preparation method thereof, and relates to the technical field of pharmaceutical preparations, and the key point of the technical scheme is that the gel ointment comprises the following preparation raw materials in parts by mass: 1 part of cepharanthine, 25 parts of a gel skeleton component, 0.4 part of a cross-linking agent, 0.01 part of an acidic pH regulator, 0.3 part of an alkaline pH regulator and 60-85 parts of a solvent; the pH (Potential of Hydrogen) value of the gel ointment is 6.5 to 8.2. Aiming at the clinical characteristics and the current treatment situation of osteoarthritis, the traditional Chinese medicine and the modern medicine dosage form are combined, and the articular cartilage protection effect of cepharanthine and the administration characteristic of the gel ointment are combined, so that the traditional Chinese medicine gel ointment for treating osteoarthritis through transdermal administration is researched and developed. And a new treatment choice is provided for patients with osteoarthritis.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and more particularly to a gel ointment for treating osteoarthritis and a preparation method thereof. Background Art

[0002] Osteoarthritis (OA) is the most common chronic musculoskeletal disease characterized by articular cartilage degradation, subchondral bone changes, osteophyte formation, low-grade synovial inflammation, and hypertrophic bone changes, leading to pain and deteriorating function. OA involves cartilage degradation and periarticular bone reactions, especially in the knee, and despite OA being one of the most common joint diseases with an increasing prevalence, the disease is difficult to treat using current therapies. Given the complexity of this pathology, there are no pharmacological treatments that can slow disease progression due to limited understanding of the pathogenesis of this disease.

[0003] In joints, tissues containing pain receptors mainly include the joint capsule, ligaments, synovium, bone, and the outer edge of the meniscus (knee joint). Inflammation lowers the threshold for pain perception; although cytokines have been evaluated as possible candidates for biochemical markers, inflammation is increasingly recognized as an important component of the pathophysiology of osteoarthritis. According to the 2000 guidelines of the American College of Rheumatology, knee osteoarthritis is a disease characterized by cartilage degeneration, and patients are usually treated with steroids, nonsteroidal anti-inflammatory drugs (NSAIDs), and cyclooxygenase-2 selective NSAIDs (such as celecoxib), which can relieve pain and inflammation but cannot restore tissue once osteoarthritis has begun.

[0004] Osteoarthritis is a chronic, progressive disease with complex mechanisms, including inflammation, periarticular bone reaction, and cartilage degradation. To date, available pain treatments have limited efficacy and are known to have associated toxicities, none of which can prevent disease progression or regenerate damaged cartilage or bone. Currently available treatment strategies aim to improve pain, provide cartilage protection or regeneration, and increase mobility. It is worth noting that some traditional Chinese medicines and Chinese herbal extracts have powerful anti-inflammatory, anti-apoptotic, and anti-catabolic activities, protecting tissues such as cartilage and synovium, providing a new option for osteoarthritis disease and its symptoms.

[0005] Gel paste refers to a patch made by mixing medicinal extracts, medicinal materials and chemical drugs with a suitable hydrophilic matrix and then spreading it on a backing material. The main components of the matrix are various water-soluble polymer materials. Gel paste has the following main characteristics:

[0006] (1) The gel ointment matrix has a large drug loading capacity, which is suitable for the use of traditional Chinese medicine and natural medicine, large doses of medication, and no side effects;

[0007] (2) Gel ointments usually contain more than 50% water, have good water retention, and can easily soften the stratum corneum of the skin, which is beneficial to the transdermal absorption of drugs;

[0008] (3) The air permeability, skin adhesion, and heat preservation of gel ointments are better than those of traditional patches and are comfortable to use;

[0009] (4) Compared with oral dosage forms, it has no first-pass effect in the liver, is not affected by gastrointestinal degradation, and has high bioavailability;

[0010] (5) Compared with injections, it is easy to use, painless, and has mild systemic side effects.

[0011] Stephania quinoline alkaloids are derived from the plant Stephania quinoline and have anti-inflammatory and analgesic activities. It is known that stephania quinoline has good anti-inflammatory activity, but there is no relevant report on whether it has activity in osteoarthritis; therefore, the present invention aims to provide a gel ointment for treating osteoarthritis and a preparation method thereof to solve the above problems. Summary of the invention

[0012] The purpose of the present invention is to provide a gel ointment for treating osteoarthritis and a preparation method thereof. The present invention combines traditional Chinese medicine with modern drug dosage forms in view of the clinical characteristics and treatment status of osteoarthritis, combines the articular cartilage protective effect of cepharanthine with the administration characteristics of the gel ointment, and develops a percutaneous Chinese medicine gel ointment for treating osteoarthritis, providing a new treatment option for patients with osteoarthritis.

[0013] The invention provides an application of stephania pine in preparing a medicine for treating osteoarthritis.

[0014] The invention also provides a pharmaceutical composition containing stephanothine for treating osteoarthritis.

[0015] The present invention is further configured as follows: the pharmaceutical composition is a gel ointment.

[0016] The present invention is further configured as follows: the pharmaceutical composition comprises the following raw materials in parts by weight:

[0017] 1 part of cepharanthine, 25 parts of gel skeleton components, 0.4 parts of cross-linking agent, 0.01 parts of acidic pH regulator, 0.3 parts of alkaline pH regulator, and 60-85 parts of solvent; the pH value of the gel ointment is 6.5-8.2.

[0018] The present invention also provides a method for preparing a pharmaceutical composition for treating osteoarthritis, comprising the following steps:

[0019] S1, mixing cepharanthine and gel skeleton components uniformly;

[0020] S2, adding a cross-linking agent and adjusting the mixture to a certain viscosity;

[0021] S3, adding an acidic pH regulator and an alkaline pH regulator to adjust the pH value;

[0022] S4, adding a solvent to make the mixture into a gel;

[0023] S5. Apply the gel-like mixture on a backing material to form a gel paste.

[0024] In summary, the present invention has the following beneficial effects:

[0025] 1. The present invention provides a novel cepharanthine gel ointment, which combines the advantages of traditional Chinese medicine and modern drug dosage forms, and provides a new treatment option for osteoarthritis patients through transdermal administration. At the same time, the cepharanthine gel ointment has anti-inflammatory and analgesic activities, and can effectively prevent the decrease in chondrocyte survival rate, lactate dehydrogenase and inflammatory cytokine release and cell apoptosis caused by IL-1β, and enhance the expression of type II collagen, thereby protecting articular cartilage and slowing down the progression of osteoarthritis;

[0026] 2. Compared with traditional oral and injection dosage forms, the gel ointment of the present invention has the advantages of being easy to use, painless, and having mild systemic side effects, which improves the patient's compliance and treatment experience. In addition, the gel ointment has no first-pass effect in the liver and is not affected by gastrointestinal degradation, so it has higher bioavailability, ensuring that the active ingredients of the drug can be better absorbed and utilized by the human body;

[0027] 3. The gel ointment matrix has a large drug loading capacity, contains more than 50% water, has good water retention, can easily soften the skin stratum corneum, and is beneficial to the transdermal absorption of drugs. At the same time, the air permeability, skin adhesion, and heat preservation of the gel ointment are better than traditional plasters, and it is more comfortable to use. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is a schematic diagram of the results of cepharanthine inhibiting the reduction of cell survival rate, LDH and inflammatory cytokine release induced by IL-1β in the examples of the present invention;

[0029] Figure 2 This is a schematic diagram of the results of stephania cerebroside attenuating IL-1β-induced chondrocyte apoptosis in an embodiment of the present invention;

[0030] Figure 3 This is a schematic diagram of the results of stephania quinata inhibiting the increase of ECM protein expression mediated by IL-1β in chondrocytes in an embodiment of the present invention;

[0031] Figure 4It is a schematic diagram of the results of stephania cerebroside regulating the SIRT1-NF-κB pathway of chondrocytes in an embodiment of the present invention. DETAILED DESCRIPTION

[0032] The following is combined with Figure 1-4 The present invention is described in further detail.

[0033] Example 1: A pharmaceutical composition containing cepharanthine for treating osteoarthritis, the pharmaceutical composition is a gel ointment, specifically comprising the following raw materials in parts by weight:

[0034] 1 part of cepharanthine, 25 parts of gel skeleton component, 0.4 parts of cross-linking agent, 0.01 parts of acidic pH regulator, 0.3 parts of alkaline pH regulator, 60-85 parts of solvent, wherein the pH value of the gel ointment is 6.5-8.2.

[0035] This embodiment preferably also includes verification that cepharanthine inhibits IL-1β-induced decrease in cell survival rate, LDH and inflammatory cytokine release, such as Figure 1 As shown, (A) is a schematic diagram of the results of MTT analysis of cell viability of chondrocytes treated with different concentrations of cepharanthine (0-100 μM) for 24 hours; (B) is a schematic diagram of the results of MTT analysis of cell viability of chondrocytes treated with 20 and 50 μM cepharanthine for 2 hours and then incubated with IL-1β (10 ng / mL) for 24 hours; (C) is a schematic diagram of the measurement of LDH in the culture medium in chondrocytes incubated with cepharanthine and IL-1β; (D) and (E) are schematic diagrams of the concentrations of inflammatory cytokines TNF-α and IL-6 in the culture medium determined by ELISA, respectively; all data are expressed as mean ± standard deviation, compared with the control group, *P<0.05, **P<0.01, ***P<0.001; compared with the IL-1β group, ##P<0.01, LDH: lactate dehydrogenase.

[0036] Figure 1 It was shown in the study that chondrocyte damage is an important pathological feature of osteoarthritis. Treatment with IL-1β leads to chondrocyte damage, which is manifested as a decrease in the number of viable cells. It is a commonly used cell model of osteoarthritis. Treatment with chelidonine can increase the number of IL-1β-treated chondrocytes, indicating that it has a potential protective effect on chondrocyte damage in osteoarthritis.

[0037] This embodiment preferably also includes verification that cepharanthine attenuates IL-1β-induced chondrocyte apoptosis, such as Figure 2As shown, (A) is a schematic diagram of the results of double staining of TUNEL and DAPI (200×); (B) is a schematic diagram of the results of calculating the percentage of TUNEL+ cells; (C) is a schematic diagram of the results of measuring the activity of Caspase-3 in chondrocytes; (D) is a schematic diagram of the results of Western blot detection of apoptosis-related proteins; (E) is a schematic diagram of the results of quantitative analysis of Bax and BCL-2; compared with the control group, **P<0.01, ***P<0.001; compared with the IL-1β group, #P<0.05.

[0038] Figure 2 It was shown in the study that chondrocyte damage is an important pathological feature of osteoarthritis. In vitro treatment of chondrocytes with IL-1β to induce apoptosis is a commonly used cell model of osteoarthritis. Treatment with chelidonine can reduce the apoptosis of IL-1β-treated chondrocytes, indicating that it has a potential protective effect on chondrocyte damage in osteoarthritis.

[0039] This embodiment preferably also includes verification that cepharanthine inhibits the increase of ECM protein expression mediated by IL-1β in chondrocytes, such as Figure 3 As shown, (A) is a schematic diagram of the results of immunofluorescence analysis to detect type II collagen (green) and staining the cell nucleus (blue) with DAPI; (B) and (C) are schematic diagrams of the results of RT-qPCR to determine the relative mRNA levels of type II collagen and aggreacan, respectively; compared with the control group, ***P<0.001; compared with the IL-1β group, ##P<0.01.

[0040] Figure 3 It is shown in the literature that cartilage is composed of chondrocytes and extracartilage matrix; type II collagen is the main collagen component in the matrix of normal articular cartilage, accounting for more than 90% of the total collagen. Therefore, the normal physiological function of type II collagen plays a decisive role in the stability of the cartilage matrix, so that the articular cartilage can obtain a certain mechanical strength, maintain uprightness, and have a certain toughness to bear the load-bearing function; in osteoarthritis, due to the degeneration and wear of cartilage, type II collagen degrades, destroying the collagen fiber framework structure, chondrocytes lose protection, and chondrocyte synthesis and metabolism are disordered, thereby causing damage to the articular cartilage; aggrecan is the main proteoglycan in the cartilage matrix. In joint diseases such as osteoarthritis, the degradation and loss of aggrecan is one of the main reasons for cartilage tissue destruction and loss of joint function. This experiment showed that IL-1β reduced the expression of type II collagen and aggrecan in chondrocytes treated with IL-1β in vitro, and chelidonine increased the expression of type II collagen and aggrecan in apoptotic chondrocytes treated with IL-1β, indicating that it has a restorative effect on collagen and proteoglycan in the cartilage matrix in osteoarthritis.

[0041] This embodiment preferably also includes stephanotis alkaloids regulating the SIRT1-NF-κB pathway in chondrocytes, such as Figure 4 As shown, (A) is a schematic diagram of the results of Western blot detection of representative protein blotting bands of SIRT1 pathway proteins; (B), (C) and (D) are schematic diagrams of the results of quantitative analysis of type II collagen, ADAMTS5 and SIRT1 proteins by Western blot, respectively; (E) is a schematic diagram of the results of Western blot detection of NF-κB p65 protein, using nuclear PCNA as an internal control; (F) is a schematic diagram of the results of the quantitative results of nuclear NF-κB p65; compared with the control group, **P<0.01, ***P<0.001; compared with the IL-1β group, #P<0.05, ##P<0.01, ###P<0.001.

[0042] Figure 4 SIRT1 is a nicotinamide adenine dinucleotide (NAD + )-dependent lysine deacetylase, which can regulate gene expression, differentiation, development and lifespan of organisms; SIRT1 expression is decreased in human and mouse knee OA cartilage as well as in the knee cartilage of aged mice, and inhibition of SIRT1 in chondrocytes leads to increased cell apoptosis; SIRT1 protein expression is downregulated in IL-1β-induced chondrocytes, as are type II collagen and ADAMTS5 expressions, which is reversed after treatment with cepharanthine, with increased SIRT1 protein expression.

[0043] This specific embodiment is merely an explanation of the present invention and is not a limitation of the present invention. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed. However, as long as they are within the scope of the claims of the present invention, they are protected by the patent law.

Claims

1. A use of cepharanthine in the preparation of a medicine for treating osteoarthritis.

2. A pharmaceutical composition containing the stephanothine for treating osteoarthritis.

3. A pharmaceutical composition for treating osteoarthritis according to claim 2, characterized in that: The pharmaceutical composition is a gel ointment.

4. A pharmaceutical composition for treating osteoarthritis according to claim 3, characterized in that: The pharmaceutical composition comprises the following raw materials in parts by weight: 1 part of cepharanthine, 25 parts of gel skeleton components, 0.4 parts of cross-linking agent, 0.01 parts of acidic pH regulator, 0.3 parts of alkaline pH regulator, and 60-85 parts of solvent; the pH value of the gel ointment is 6.5-8.

2.

5. The method for preparing a pharmaceutical composition for treating osteoarthritis according to claim 4, characterized in that: The following steps are involved: S1, mixing cepharanthine and gel skeleton components uniformly; S2, adding a cross-linking agent and adjusting the mixture to a certain viscosity; S3, adding an acidic pH regulator and an alkaline pH regulator to adjust the pH value; S4, adding a solvent to make the mixture into a gel; S5. Apply the gel-like mixture on a backing material to form a gel paste.

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