Use of Compound C692-0434 in Treating Osteoarthritis

Abstract

The present invention relates to the use of compound C692-0434 in the treatment of osteoarthritis. By screening existing library compounds, the present invention has discovered the excellent properties of compound C692-0434 in the treatment of osteoarthritis. It can not only increase the proteoglycan content of OA chondrocytes, but also significantly increase the content of the extracellular matrix of chondrocytes in OA rats, reduce cartilage damage, and alleviate the condition of OA rats, making it suitable for development as a new generation of anti-osteoarthritis drugs.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceuticals. Specifically, the present invention relates to the use of compound C692-0434 for treating osteoarthritis. Background Art

[0002] Osteoarthritis (OA) is a common chronic degenerative disease, and its occurrence and development are related to various factors such as acute and chronic joint injuries, age, obesity, and metabolic bone diseases. The clinical features 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, the main methods are drug treatment and acupuncture physiotherapy 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, the drug treatments for OA mainly include non-steroidal anti-inflammatory drugs, glucocorticoids, and glycosaminoglycans. Among them, non-steroidal anti-inflammatory drugs are the most common treatment method in clinical practice. By inhibiting the synthesis of prostaglandins and directly acting on nociceptors, they reduce the formation and release of pain-causing substances, thereby exerting anti-inflammatory and analgesic effects. Currently, commonly used drugs include the oral drug celecoxib [Puljak L, et.al. Celecoxib for osteoarthritis. Cochrane Database SystRev. 2017May22;5(5):CD009865], the topical and intra-articular injection of diclofenac [Hagen M, et.al. Skinpenetration and tissue permeation after topical administration ofdiclofenac. Curr Med Res Opin. 2017Sep;33(9):1623-1634; Nishida Y, et.Al. Efficacyand Safety of Diclofenac-Hyaluronate Conjugate(Diclofenac Etalhyaluronate)forKnee 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 theManagement of Osteoarthritis. An Opinion-Based Report from the ESCEO. DrugsAging. 2016Feb;33(2):75-85]. Diacerein alleviates oxidative stress and osteoarthritis in a mouse model 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 application of non-steroidal 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 digestive tract diseases, or those taking hormones / anticoagulants in combination; in terms of the cardiovascular system, long-term high-dose use may increase the risks of hypertension, heart failure, myocardial infarction, and stroke. In addition, non-steroidal anti-inflammatory drugs may reduce renal blood flow by inhibiting prostaglandins, leading to acute or chronic kidney injury, water and sodium retention, or hyperkalemia. Elderly, diabetic, or nephropathy patients need to regularly monitor renal function. Other potential risks include elevated liver enzymes (alert for hepatotoxicity), platelet inhibition and bleeding tendency caused by non-selective non-steroidal anti-inflammatory drugs, iron-deficiency anemia caused by occult gastrointestinal bleeding, as well as headache, dizziness, or allergic reactions, etc.

[0005] More importantly, OA is not simply "joint wear", but a multi-factorial disease involving cartilage degeneration, synovial inflammation, abnormal bone remodeling, and metabolic imbalance. Traditional drugs can only relieve pain symptomatically and cannot delay the disease progression or repair damaged tissues. In addition, some patients have poor responses to existing drugs or have contraindications (such as severe cardiovascular diseases, peptic ulcers), and there is an urgent need for a safer alternative. Therefore, there is an urgent need to develop drugs that can fundamentally improve the cartilage degeneration of OA and relieve or even reverse the disease 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 the use of compound C692-0434 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating osteoarthritis, and the compound has the following structure:

[0008]

[0009] The English name of the said 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 proinflammatory cytokine-induced osteoarthritis. 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 salts include salts formed by the compound and pharmaceutically acceptable acids. 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 its pharmaceutically acceptable salts.

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

[0015] As a pharmaceutically acceptable adjuvant, any adjuvant known to be suitable for preparing a specific drug can be used. Examples thereof include, but are not limited to, 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. Adjuvants of the type suitable for the desired formulation and the desired mode of administration are particularly 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 form suitable for oral administration includes tablets, lozenges, hard or soft gelatin capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs.

[0018] The form suitable for administration by insufflation includes fine powders or liquid aerosols.

[0019] The form suitable for parenteral administration includes 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 and disintegrating agents 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 p-hydroxybenzoate 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 adjuvants for hard gelatin capsules include inert solid diluents such as calcium carbonate, calcium phosphate or kaolin; suitable pharmaceutically acceptable adjuvants for soft gelatin capsules include water or oils such as peanut oil, liquid paraffin or olive oil.

[0022] Aqueous suspensions generally contain the active ingredient in finely powdered form and one or more suspending agents such as sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, methyl cellulose, polyvinylpyrrolidone and gum arabic; dispersing or wetting agents such as lecithin or condensates of alkylene oxides with fatty acids or condensates of ethylene oxide with long-chain fatty alcohols, such as heptadecaethyleneoxycetanol, condensates of ethylene oxide with partial esters derived from fatty acids and hexitol, such as polyoxyethylene sorbitan monooleate or condensates of ethylene oxide with partial esters derived from fatty acids and hexitanhydrides such as polyoxyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, coloring agents, antioxidants, flavoring agents and / or sweetening agents.

[0023] Oil suspensions may 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 thickening agents such as beeswax, solid paraffin or cetyl alcohol. Sweetening agents and flavoring agents may also be added to obtain a palatable oral preparation. Antioxidants such as ascorbic acid may also be added to preserve these compositions.

[0024] The emulsion may be in the form of an oil-in-water type. The oil phase may 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 may be, for example, natural gums such as gum arabic or tragacanth, natural phospholipids such as soy lecithin, esters or partial esters derived from fatty acids and sorbitan (such as sorbitan monooleate) and condensates of said partial esters and ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners, flavoring agents and preservatives.

[0025] Dispersible powders and granules suitable for the preparation of aqueous suspensions by the addition of water generally contain the active ingredient as well as a dispersing or wetting agent, a suspending agent and one or more preservatives. Examples of suitable dispersing 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 medicament suitable for administration by insufflation may be in the form of a fine powder containing particles with an average 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 medicament suitable for administration by insufflation may also be in the form of a pressurized aerosol conventionally used to disperse the active ingredient into an aerosol containing fine solid particles or droplets. Conventional aerosol propellants such as volatile fluorinated hydrocarbons or hydrocarbons may be used, and an aerosol device is conveniently used to dispense a metered amount of the active ingredient.

[0027] In the form of an aqueous or oily suspension for injection, which may be formulated according to known methods using one or more suitable dispersing or wetting agents and suspending agents as described above. The sterile injectable preparation may 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 Chapter 25.2, Volume 5 of Comprehensive Medicinal Chemistry (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 a formulation to reduce the dosage and the uniformity of the 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 judgment. The specific effective dose level for any particular patient or organism will depend on many factors including the disorder being treated and the severity of the disorder, the activity of the specific compound, the specific composition used, the patient's age, weight, health status, gender and eating habits, the time of administration, the route of administration and the excretion rate of the specific compound used, the duration of treatment, the drug application in combination therapy or in combination with a compound with special effects, and other factors well known in the pharmaceutical field.

[0030] Beneficial effects

[0031] The present invention relates to the use of compound C692-0434 in the treatment of osteoarthritis. By screening the existing library compounds, the present invention discovers the excellent properties of compound C692-0434 in the treatment of osteoarthritis. It can increase the proteoglycan content of OA chondrocytes and improve the mRNA levels of cartilage-related marker factors Col2a1, Acan and Sox9 in OA chondrocytes; it can significantly increase the content of the extracellular matrix of chondrocytes in OA rats induced by TNF-α and the content of the extracellular matrix of chondrocytes in ACLT rats, reduce cartilage damage and greatly relieve the condition of OA rats; moreover, the compound has high safety. Therefore, it is suitable for developing as a new generation of drugs for the treatment of osteoarthritis. Description of the drawings

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

[0033] Figure 2 : Compound C692-0434 significantly increases the mRNA levels of cartilage-related marker factors in rat OA chondrocytes; * indicates P<0.05, ** indicates P<0.01;

[0034] Figure 3 : Compound C692-0434 significantly increases the content of the extracellular matrix of chondrocytes;

[0035] Figure 4 : Compound C692-0434 alleviates the condition of rat OA;

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

[0037] Figure 6 : Micro-CT images of OA rats and the treatment situation. Detailed implementation manners

[0038] The present invention is described in more detail below 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 construed as having the meanings defined in the dictionary, but should be understood as having meanings consistent with their meanings in the context of the present invention based on the following principles: the concept of the terms 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 those described herein can also be used for the practice or testing of the present invention. The materials, methods, and examples disclosed herein are illustrative only and are not intended to be limiting.

[0040] Through various biological experiments, it has been observed that compound C692-0434 has a significant effect on improving the condition of osteoarthritis 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 Alcian blue staining was performed. It was observed that the small molecule compound C692-0434 significantly increased the content of proteoglycan ( Figure 1 ). The above-mentioned rat OA cells co-cultured with compound C692-0434 were subjected to RT-PCR analysis, and it was found that the mRNA levels of cartilage-related marker factors Col2a1, Acan, and Sox9 in rat OA chondrocytes were significantly increased ( Figure 2 , n = 3 for each group).

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

[0044] TNF-α was injected into the joint cavity of normal rats to make them OA model mice. Different concentrations (100 nM, 1 μM, 10 μM) of C692-0434 were injected into the knee joint cavity of the model mice. After 4 weeks, the samples were taken, fixed, decalcified, histologically stained, and 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 chondrocytes in the experimental group was significantly increased compared with the negative control group, indicating an increase in the synthesis of the extracellular matrix of chondrocytes and a significant reduction in the degree of cartilage damage. The condition of rat OA was alleviated and recovered. Among them, 1 μM of compound C692-0434 significantly increased the content of the extracellular matrix of chondrocytes, which was closest to the content of the extracellular matrix of chondrocytes in the normal group mice (Figure 3 )。

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

[0046] Anterior cruciate ligament transection was performed on normal rats to make them OA model rats. 200 μL of compound C692-0434 with a concentration of 1 μM was injected into the knee joint cavity of ACLT model rats. After 4 weeks, the samples were taken, fixed, decalcified, and histologically stained, and 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 chondrocytes in the experimental group was significantly increased compared with the negative control group, indicating an increase in the synthesis of the extracellular matrix of chondrocytes and a significant reduction in the degree of cartilage damage. The OA condition of the rats was alleviated and recovered ( Figure 4 ). And a quantitative analysis - OARSI score was made for the results of the above histological staining. It was found that after intra-articular injection of compound C692-0434 to the OA model rats of ACLT, their OARSI scores were significantly decreased and the OA condition of the rats was alleviated ( Figure 5 ). The 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 kidneys of rats treated with compound C692-0434 by knee injection. It was found that the tissue and cell morphological structures of the above organs were all normal, and no toxicity of compound C692-0434 was observed, preliminarily verifying the biosafety of compound C692-0434.

[0049] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the method of the present invention, several improvements and supplements can be made, and these improvements and supplements should also be regarded as the protection scope of the present invention.

Claims

1. Use of a compound C692-0434 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating osteoarthritis, said compound having the following structure: 。 2. The application according to claim 1, characterized in that The osteoarthritis is proinflammatory cytokine-induced osteoarthritis.

3. The application according to claim 2, wherein The proinflammatory cytokines include TNF-α, IL-1β, IL-6, IL-17, IL-18, IL-8.

4. The application according to claim 1, wherein The osteoarthritis is anterior cruciate ligament transection-induced osteoarthritis.

5. The application according to claim 1, characterized in that, The drug further comprises a pharmaceutically acceptable adjuvant.

6. The application according to claim 5, wherein The pharmaceutically acceptable adjuvants include solvents, excipients, dispersants, emulsifiers, solubilizers, gel formers, ointment bases, antioxidants, preservatives, stabilizers, fillers, binders, thickeners, complexing agents, disintegrants, buffers, penetration enhancers, polymers, lubricants, coating agents, propellants, tonicity regulators, surfactants, colorants and flavoring agents.

7. The application according to claim 5, characterized in that, The pharmaceutically acceptable adjuvants include sweeteners and dyes.

8. The application according to claim 1, characterized in that, The drug is in a form suitable for oral administration, a form suitable for inhalation administration, or a form suitable for parenteral administration.

9. The application according to claim 8, wherein The form suitable for oral administration includes tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs; the form suitable for inhalation administration includes fine powders or liquid aerosols; the form suitable for parenteral administration includes sterile aqueous or oily solutions for intravenous, subcutaneous or intramuscular administration or suppositories for rectal administration.

Images