Anti-osteoarthritis application of compound C692-0434

Abstract

The invention relates to an application of a compound C692-0434 in resisting osteoarthritis. Existing library compounds are screened, the excellent property of the compound C692-0434 in resisting osteoarthritis is found, the content of proteoglycan of OA cartilage cells can be increased, the content of cartilage extracellular matrixes of OA rats can be obviously increased, cartilage injuries are reduced, the disease condition of the OA rats is relieved, and the compound C692-0434 can be used for resisting osteoarthritis. The compound is suitable for being used for developing a new generation of anti-osteoarthritis medicine.

Description

Technical Field

[0001] The present invention belongs to the field of medicines, and in particular, the present invention relates to the anti-osteoarthritis use of compound C692-0434. Background Art

[0002] Osteoarthritis (OA) is a common chronic degenerative disease, the development of which is related to multiple factors such as acute and chronic joint injuries, age, obesity and metabolic bone disease. The clinical characteristics of OA include pain, joint stiffness, joint hypertrophy, limited movement, muscle weakness, local tenderness, deformity and loss of function, which seriously affect the quality of life of elderly patients and become the main cause of disability in this population. For early OA, drug therapy and acupuncture and physical therapy are mainly used to relieve joint pain and delay the progression of OA; for late OA, joint replacement surgery can be used to restore joint function.

[0003] Currently, drug treatments for OA mainly include nonsteroidal anti-inflammatory drugs, glucocorticoids, and glycosaminoglycans. Among them, nonsteroidal anti-inflammatory drugs are the most important treatment method in clinical practice. They inhibit the synthesis of prostaglandins and directly act on nociceptors, reducing the formation and release of pain-causing substances, thereby exerting anti-inflammatory and analgesic effects. Currently, commonly used drugs include oral celecoxib [Puljak L, et.al. Celecoxib for osteoarthritis. Cochrane Database Syst Rev. 2017 May 22; 5(5): CD009865], topical and intra-articular injection of diclofenac [Hagen M, et.al. Skin penetration and tissue permeation after topical administration of diclofenac. Curr Med Res Opin. 2017 Sep; 33(9): 1623-1634; Nishida Y, et.al. Efficacy and Safety of Diclofenac-Hyaluronate Conjugate (Diclofenac Etalhyaluronate) for Knee Osteoarthritis: A Randomized Phase III Trial in Japan.ArthritisRheumatol.2021Sep;73(9):1646-1655], etc. Diacerein controls and improves the condition of osteoarthritis by interfering with interleukin-1, and has anti-inflammatory, anti-catabolic and anabolic properties on cartilage and synovium [Pavelka K, BruyèreO, Cooper C, Kanis JA, Leeb BF, Maheu E, Martel-Pelletier J, Monfort J, PelletierJP, Rizzoli R, Reginster JY.Diacerein: Benefits, Risks and Place in the Management of Osteoarthritis. An Opinion-Based Report from the ESCEO. DrugsAging.2016Feb;33(2):75-85], Diacerein alleviates oxidative stress and osteoarthritis in mouse models by inhibiting peroxisome proliferator-activated receptor-γ [Chen X, et al.Reversal of EpigeneticPeroxisome Proliferator-Activated Receptor-γSuppression by DiacereinAlleviates Oxidative Stress and Osteoarthritis in Mice. Antioxid RedoxSignal. 2022Jul; 37(1-3):40-53]. .

[0004] However, long-term use of nonsteroidal anti-inflammatory drugs has many side effects. Gastrointestinal damage is the most common risk, manifested as gastric mucosal erosion, ulcers, and even bleeding or perforation, especially in the elderly, those with a history of gastrointestinal diseases, or those taking hormones / anticoagulants; in terms of the cardiovascular system, long-term high-dose use may increase the risk of hypertension, heart failure, myocardial infarction, and stroke. In addition, nonsteroidal anti-inflammatory drugs may reduce renal blood flow by inhibiting prostaglandins, leading to acute or chronic renal damage, water and sodium retention, or hyperkalemia. Elderly patients, patients with diabetes or kidney disease need to monitor renal function regularly. Other potential risks include elevated liver enzymes (beware of liver toxicity), platelet inhibition and bleeding tendency caused by non-selective nonsteroidal anti-inflammatory drugs, iron deficiency anemia caused by occult gastrointestinal bleeding, and headaches, dizziness, or allergic reactions.

[0005] More importantly, OA is not simply "joint wear and tear", but a multifactorial disease involving cartilage degeneration, synovial inflammation, abnormal bone remodeling and metabolic imbalance. Traditional drugs can only provide symptomatic analgesia but cannot delay disease progression or repair damaged tissues. In addition, some patients do not respond well to existing drugs or have contraindications (such as severe cardiovascular disease and gastrointestinal ulcers), and are in urgent need of safer alternatives. Therefore, there is an urgent need to develop drugs that can fundamentally improve the degenerative changes in OA cartilage and alleviate or even reverse the progression of OA. Summary of the invention

[0006] In order to solve the problems existing in the prior art, the present invention screened the existing library compounds and obtained a compound that can effectively treat osteoarthritis, namely compound C692-0434.

[0007] In a first aspect, the present invention provides a use of a compound C692-0434 or a pharmaceutically acceptable salt thereof in the preparation of an anti-osteoarthritis drug, wherein the compound has the following structure:

[0008]

[0009] The English name of the compound is 8-((4-(4-fluorophenyl)piperazin-1-yl)methyl)-3-methyl-7-(3-phenylpropyl)-3,7-dihydro-1H-purine-2,6-dione, the Chinese name is 8-((4-(4-fluorophenyl)piperazin-1-yl)methyl)-3-methyl-7-(3-phenylpropyl)-3,7-dihydro-1H-purine-2,6-dione, the Cas registration number is 847409-19-2, and the number in ChemDiv is C692-0434.

[0010] In one embodiment, the osteoarthritis is osteoarthritis induced by proinflammatory cytokines. Preferably, the proinflammatory cytokines include TNF-α, IL-1β, IL-6, IL-17, IL-18, IL-8.

[0011] In one embodiment, the osteoarthritis is anterior cruciate ligament transection-induced osteoarthritis.

[0012] In one embodiment, the pharmaceutically acceptable salt includes a salt formed by a compound and a pharmaceutically acceptable acid. Suitable acids include pharmaceutically acceptable inorganic acids and organic acids. Representative pharmaceutically acceptable acids include hydrogen chloride, hydrogen bromide, nitric acid, sulfuric acid, sulfonic acid, phosphoric acid, acetic acid, glycolic acid, phenylacetic acid, propionic acid, butyric acid, valeric acid, maleic acid, acrylic acid, fumaric acid, succinic acid, malic acid, malonic acid, tartaric acid, citric acid, salicylic acid, benzoic acid, tannic acid, formic acid, stearic acid, lactic acid, ascorbic acid, methanesulfonic acid, p-toluenesulfonic acid, oleic acid, lauric acid, etc.

[0013] In a second aspect, the present invention also provides a pharmaceutical composition for anti-osteoarthritis comprising compound C692-0434 or a pharmaceutically acceptable salt thereof.

[0014] In one embodiment, the medicament or pharmaceutical composition further comprises a pharmaceutically acceptable adjuvant.

[0015] As pharmaceutically acceptable adjuvants, any adjuvant known to be suitable for preparing a specific drug can be used. Examples include, but are not limited to, solvents, excipients, dispersants, emulsifiers, solubilizers, gel formers, ointment bases, antioxidants, preservatives, stabilizers, carriers, fillers, bonding agents, thickeners, complexing agents, disintegrants, buffers, penetration enhancers, polymerizers, lubricants, coating agents, propellants, tonicity regulators, surfactants, colorants, flavoring agents, sweeteners and dyes. Particularly adjuvants of the type suitable for the desired formulation and the desired mode of administration are used.

[0016] The medicament or pharmaceutical composition of the present invention may be in a form suitable for oral administration, a form suitable for administration by inhalation, or a form suitable for parenteral administration.

[0017] The forms suitable for oral administration include tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs.

[0018] The forms suitable for administration by insufflation include fine powders or liquid aerosols.

[0019] The forms suitable for parenteral administration include sterile aqueous or oily solutions for intravenous, subcutaneous or intramuscular administration or suppositories for rectal administration.

[0020] Suitable pharmaceutically acceptable adjuvants for tablets include, for example, inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate; granulating agents and disintegrants such as corn starch or alginic acid; binders such as starch; lubricants such as magnesium stearate, stearic acid or talc; preservatives such as ethyl or propyl paraben and antioxidants such as ascorbic acid. Tablets may be uncoated or coated to improve their disintegration in the gastrointestinal tract and subsequent absorption of the active ingredient or to improve their stability and / or appearance, in either case using conventional coating agents and methods well known in the art.

[0021] Suitable pharmaceutically acceptable excipients for hard capsules include inert solid diluents such as calcium carbonate, calcium phosphate or kaolin; suitable pharmaceutically acceptable excipients for soft capsules include water or oils such as peanut oil, liquid paraffin or olive oil.

[0022] Aqueous suspensions usually contain active ingredients in fine powder form and one or more suspending agents such as sodium carboxymethylcellulose, hydroxypropyl methylcellulose, sodium alginate, methylcellulose, polyvinyl pyrrolidone and gum arabic; dispersants or wetting agents such as lecithin or condensates of alkylene oxide and fatty acids or condensates of ethylene oxide and long-chain fatty alcohols, such as 17-decane ethyleneoxy hexadecyl alcohol, condensates of ethylene oxide and partial esters derived from fatty acids and hexitol, such as polyoxyethylene sorbitol monooleate or condensates of ethylene oxide and partial esters derived from fatty acids and hexitol anhydrides such as polyoxyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, colorants, antioxidants, flavorings and / or sweeteners.

[0023] Oil suspensions can be prepared by suspending the active ingredient in a vegetable oil (such as peanut oil, olive oil, sesame oil or coconut oil) or a mineral oil (such as liquid paraffin). Oil suspensions may also contain thickeners such as beeswax, paraffin wax or cetyl alcohol. Sweeteners and flavoring agents may also be added to obtain a palatable oral preparation. These compositions may also be preserved by adding antioxidants such as ascorbic acid.

[0024] The emulsion can be in the form of oil-in-water type. The oil phase can be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin, or any mixture thereof. Suitable emulsifiers can be, for example, natural gums such as gum arabic or gum tragacanth, natural phospholipids such as soybean, lecithin, esters or partial esters (such as sorbitan monooleate) derived from fatty acids and hexitol anhydrides, and condensates of the partial esters and ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsion can also include sweeteners, flavorings and preservatives.

[0025] Dispersible powders and granules suitable for preparing aqueous suspensions by adding water generally contain the active ingredient together with a dispersant or wetting agent, a suspending agent and one or more preservatives. Examples of suitable dispersants or wetting agents and suspending agents are those mentioned above. Other excipients such as sweeteners, flavoring agents and coloring agents may also be present.

[0026] The medicine suitable for insufflation can be in the form of a fine powder containing particles with a mean diameter of, for example, 30 μm or less, which powder itself contains only the active ingredient or contains the active ingredient diluted with one or more physiologically acceptable carriers such as lactose. The medicine suitable for insufflation can also be in the form of a conventional pressurized aerosol for dispersing the active ingredient into an aerosol containing fine solid particles or droplets. Conventional aerosol propellants such as volatile fluorocarbons or hydrocarbons can be used, and aerosol devices are conveniently used to distribute metered amounts of active ingredients.

[0027] Injectable water or oil suspension form, which can be prepared according to known methods using one or more suitable dispersants or wetting agents and suspending agents as described above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol.

[0028] For more information on formulations, see Comprehensive Medicinal Chemistry, Vol. 5, Chapter 25.2 (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990, which is incorporated herein by reference.

[0029] The pharmaceutical composition of the present invention is preferably prepared into a dosage unit type according to the formulation formula to reduce the uniformity of the dosage and dosage. It should be understood that the total daily usage of the drug or pharmaceutical composition of the present invention will be determined by the attending physician based on a reliable medical range of judgment. The specific effective dosage level for any particular patient or organism will depend on many factors including the severity of the disease and the disease being treated, the activity of the specific compound, the specific composition used, the patient's age, weight, health, sex and dietary habits, the time of administration, the route of administration and the excretion rate of the specific compound used, the duration of treatment, the drug is used in combination or in combination with a specific compound, and other factors known to the pharmaceutical field.

[0030] Beneficial Effects

[0031] The present invention relates to the use of compound C692-0434 in the treatment of osteoarthritis. The present invention screens existing library compounds and finds that compound C692-0434 has excellent properties in the treatment of osteoarthritis, which can increase the proteoglycan content of OA chondrocytes and improve the mRNA levels of cartilage-related markers Col2a1, Acan and Sox9 in OA chondrocytes; it can significantly increase the content of cartilage extracellular matrix in TNF-α-induced OA rats and the content of cartilage extracellular matrix in ACLT rats, reduce cartilage damage, and greatly alleviate the condition of OA rats; and the compound has high safety. Therefore, it is suitable for the development of a new generation of anti-osteoarthritis drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 : Compound C692-0434 increased the proteoglycan content of OA chondrocytes;

[0033] Figure 2 : Compound C692-0434 induced a significant increase in the mRNA levels of cartilage-related markers in rat OA chondrocytes; * indicates P < 0.05, ** indicates P < 0.01;

[0034] Figure 3 : Compound C692-0434 significantly increased the content of extracellular matrix in cartilage cells;

[0035] Figure 4 : Compound C692-0434 alleviates OA symptoms in rats;

[0036] Figure 5 : Compound C692-0434 reduced the OARSI score of OA rats; *** indicates P < 0.001;

[0037] Figure 6 : Micro-CT images of OA rats and treatment conditions. DETAILED DESCRIPTION

[0038] The present invention is described in more detail hereinafter to facilitate understanding of the present invention.

[0039] It should be understood that the terms or words used in the specification and claims should not be understood to have the meaning defined in the dictionary, but should be understood to have the meaning consistent with its meaning in the context of the present invention on the basis of the following principles: the concept of the term can be appropriately defined by the inventor for the best description of the present invention. Preferred methods and materials are described below, but methods and materials similar or equivalent to the methods and materials described herein can also be used in the practice or testing of the present invention. The materials, methods and examples disclosed herein are only illustrative and are not intended to be limiting.

[0040] Through various biological experiments, it has been observed that compound C692-0434 has a significant therapeutic effect on improving osteoarthritis symptoms and controlling the progression of osteoarthritis.

[0041] Example 1: In vitro anti-osteoarthritis effect of compound C692-0434

[0042] The small molecule compound C692-0434 with a final concentration of 1 μM was co-cultured with TNF-α-induced rat OA chondrocytes for 5 days and stained with Alcian blue. It was observed that the small molecule compound C692-0434 significantly increased the content of proteoglycan ( Figure 1 ). RT-PCR analysis was performed on the rat OA cells co-cultured with compound C692-0434, and it was found that the mRNA levels of cartilage-related markers Col2a1, Acan and Sox9 in rat OA chondrocytes were significantly increased ( Figure 2 , n=3 per group).

[0043] Example 2: Validation of in vivo experimental animal model (TNF-α-induced OA model mouse)

[0044] Normal rats were intra-articularly injected with TNF-α to make them OA model mice. 200μL of C692-0434 of different concentrations (100nM, 1μM, 10μM) was injected into the knee joint cavity of the model mice. After 4 weeks, the samples were fixed, decalcified, and histologically stained. Histological analysis was performed under a microscope and compared with the normal group and the negative control group. The experiment found that the content of the extracellular matrix of the cartilage cells in the experimental group was significantly increased compared with that in the negative control group, reflecting an increase in the synthesis of the extracellular matrix of the cartilage cells and a significant decrease in the degree of cartilage damage. The OA condition of the rats was alleviated and recovered. Among them, 1μM of the compound C692-0434 significantly increased the content of the extracellular matrix of the cartilage cells, which was closest to the content of the extracellular matrix of the cartilage cells of the normal group of mice ( Figure 3 ).

[0045] Example 3: Validation of in vivo experimental animal model (OA model mice induced by anterior cruciate ligament rupture)

[0046] Normal rats underwent anterior cruciate ligament rupture to make them OA model mice. 200 μL of 1 μM compound C692-0434 was injected into the knee joint cavity of ACLT model mice. After 4 weeks, the samples were fixed, decalcified, and histologically stained. Histological analysis was performed under a microscope and compared with the normal group and negative control group. The experiment found that the content of cartilage extracellular matrix in the experimental group was significantly increased compared with that in the negative control group, reflecting an increase in the synthesis of cartilage extracellular matrix and a significant decrease in the degree of cartilage damage. The OA condition of the rats was alleviated and recovered ( Figure 4 ). A quantitative analysis of the above histological staining results was performed - OARSI score. It was found that after intra-articular injection of compound C692-0434 in ACLT OA model mice, their OARSI scores were significantly reduced, and the OA condition of the rats was alleviated ( Figure 5 ). Micro-CT imaging evidence for this in vivo experiment also showed the same result ( Figure 6 ).

[0047] Example 4: Biosafety

[0048] After the in vivo experiment was completed, HE staining was performed on the liver, heart, spleen and kidney of rats treated with knee joint injection of compound C692-0434. It was found that the tissue and cell morphology of the above organs were normal, and no toxicity of compound C692-0434 was observed, which preliminarily verified the biosafety of compound C692-0434.

[0049] The above is only a preferred embodiment of the present invention. It should be pointed out that ordinary technicians in this technical field can make several improvements and supplements without departing from the method of the present invention. These improvements and supplements should also be regarded as the scope of protection of the present invention.

Claims

1. Use of a compound C692-0434 or a pharmaceutically acceptable salt thereof in the preparation of an anti-osteoarthritis drug, wherein the compound has the following structure:

2. The use according to claim 1, characterized in that: The osteoarthritis is pro-inflammatory cytokine-induced osteoarthritis.

3. The use according to claim 2, characterized in that: The proinflammatory cytokines include TNF-α, IL-1β, IL-6, IL-17, IL-18, and IL-8.

4. The use according to claim 1, characterized in that: The osteoarthritis is anterior cruciate ligament transection-induced osteoarthritis.

5. The use according to claim 1, characterized in that: The medicament further comprises a pharmaceutically acceptable adjuvant.

6. The use according to claim 5, characterized in that: The pharmaceutically acceptable adjuvants include solvents, excipients, dispersants, emulsifiers, solubilizers, gel formers, ointment bases, antioxidants, preservatives, stabilizers, carriers, fillers, binders, thickeners, complexing agents, disintegrants, buffers, penetration enhancers, polymerizers, lubricants, coating agents, propellants, tonicity regulators, surfactants, colorants, flavoring agents, sweeteners and dyes.

7. The use according to claim 1, characterized in that: The medicament is in a form suitable for oral administration, a form suitable for administration by inhalation, or a form suitable for parenteral administration.

8. The use according to claim 7, characterized in that: The form suitable for oral administration includes tablets, lozenges, hard or soft capsules, water or oil suspensions, emulsions, dispersible powders or granules, syrups or elixirs; the form suitable for insufflation administration includes fine powders or liquid aerosols; the form suitable for parenteral administration includes sterile water or oil solutions for intravenous, subcutaneous or intramuscular administration or suppositories for rectal administration.

9. A pharmaceutical composition for anti-osteoarthritis comprising compound C692-0434 or a pharmaceutically acceptable salt thereof, wherein the compound has the following structure:

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