Drug for treating osteoarthritis

Abstract

Provided is a drug for treating osteoarthritis. Specifically, the present invention provides for the first time a use of annexin A2 in the preparation of a pharmaceutical composition for preventing or treating osteoarthritis.

Description

A drug for treating osteoarthritis Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a medicine for treating osteoarthritis. Background Art

[0002] Osteoarthritis (OA) is the most common joint disease characterized by degeneration of articular cartilage. OA primarily affects the primary weight-bearing joints, including the hip and knee. Clinical manifestations of OA include joint pain, swelling, limited mobility, and functional impairment. Tissue changes within OA joints manifest as cartilage matrix destruction and loss, osteophyte formation, subchondral bone sclerosis, and sterile synovial inflammation. The development of OA is influenced by multiple mechanisms, with genetics, trauma, age, and obesity being the primary factors. From 2005 to 2015, the prevalence of knee OA increased by 32.7%, making OA one of the leading causes of years of disability worldwide. Furthermore, OA contributes to an annual economic burden of at least US$89.1 billion, representing 1% to 2.5% of GDP in high-income countries. However, despite the increasing incidence and severity of OA, there is currently no cure or effective disease-modifying therapy due to a lack of understanding of the disease's pathogenesis. Current pharmacological treatments for OA, such as corticosteroids, hyaluronic acid, and opioids, primarily alleviate symptoms but fail to halt or reverse cartilage degeneration. Furthermore, these drugs have side effects and toxicity. Due to irreversible disease progression, most patients ultimately require joint replacement, which imposes a significant societal and economic burden. These limitations require the development of novel therapies that, in addition to their anti-inflammatory and analgesic effects, can improve cartilage structure and reverse cartilage destruction to improve overall joint function while reducing side effects. However, such treatments currently do not exist. Therefore, there is an urgent need to identify effective drugs to slow the progression of OA and to investigate the underlying molecular mechanisms.

[0003] Summary of the Invention

[0004] The purpose of the present invention is to provide a medicine for treating osteoarthritis.

[0005] In a first aspect, the present invention provides use of Annexin A2 in preparing a pharmaceutical composition for preventing or treating osteoarthritis.

[0006] In another preferred embodiment, the osteoarthritis includes articular cartilage degeneration damage, reactive hyperplasia of joint margin and / or subchondral bone caused by aging, obesity, strain, trauma and / or congenital abnormalities of the joints.

[0007] In another preferred embodiment, the osteoarthritis is selected from the group consisting of cervical osteoarthritis, lumbar osteoarthritis, knee osteoarthritis, hip osteoarthritis, or a combination thereof.

[0008] In another preferred embodiment, the osteoarthritis includes knee osteoarthritis, which is a chronic osteoarthritis characterized by degenerative changes in the knee joint cartilage.

[0009] In another preferred embodiment, the prevention or treatment of arthritis is carried out by one or more methods selected from the following groups:

[0010] (a) Reduce cartilage damage;

[0011] (b) promote chondrocyte proliferation;

[0012] (c) Reduce inflammation in joints (including synovium);

[0013] (d) Relieve joint pain.

[0014] In another preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0015] In another preferred embodiment, the pharmaceutical composition is in the form of an oral preparation, an external preparation or an injection preparation.

[0016] In another preferred embodiment, the injection preparation is an intravenous injection or an intramuscular injection.

[0017] In another preferred embodiment, the dosage form of the composition or preparation is a solid dosage form, a semisolid dosage form, or a liquid dosage form, such as a tablet, a capsule, a microcapsule, a powder injection, an oral solution, an injection, a solution, a gel, an emulsion, an ointment, a cream, a paste, a powder, a patch or a lozenge.

[0018] In another preferred embodiment, the composition or preparation is administered externally, topically, or by subcutaneous injection.

[0019] In a second aspect, the present invention provides a pharmaceutical composition comprising (a) Annexin A2; and (b) a pharmaceutically acceptable carrier.

[0020] In another preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0021] In another preferred embodiment, the pharmaceutical composition is in the form of an oral preparation, an external preparation or an injection preparation.

[0022] In another preferred embodiment, the injection preparation is an intravenous injection or an intramuscular injection.

[0023] In another preferred embodiment, the dosage form of the composition or preparation is a solid dosage form, a semisolid dosage form, or a liquid dosage form, such as a tablet, a capsule, a microcapsule, a powder injection, an oral solution, an injection, a solution, a gel, an emulsion, an ointment, a cream, a paste, a powder, a patch or a lozenge.

[0024] In another preferred embodiment, the composition or preparation is administered externally, topically, or by subcutaneous injection.

[0025] In another preferred embodiment, in the pharmaceutical composition, the mass percentage of annexin A2 is 0.01 wt%-99 wt%, preferably 0.1-50 wt%, 1-20 wt%, based on the total weight of the composition.

[0026] In a third aspect, the present invention provides a method for preventing or treating osteoarthritis, comprising administering Annexin A2 or the composition of the second aspect of the present invention to a subject in need thereof.

[0027] In another preferred embodiment, the subject is a human or non-human mammal.

[0028] In another preferred embodiment, the non-human mammals include rodents, such as rats and mice.

[0029] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1. Promotional effect of Annexin A2 on chondrocyte proliferation.

[0031] Figure 2 Protective effect of Annexin A2 on chondrocytes.

[0032] Figure 3 Annexin A2 treatment alleviates joint pain symptoms.

[0033] Figure 4 Annexin A2 inhibits inflammatory response in joint synovium.

[0034] Figure 5 Annexin A2 protects cartilage from damage. DETAILED DESCRIPTION

[0035] After extensive and in-depth research, numerous screenings, and tests, the present inventors have provided, for the first time, the use of Annexin A2 in preventing or treating osteoarthritis, on which basis the present invention was completed.

[0036] the term

[0037] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0038] As used herein, the terms "comprising" or "including" may be open, semi-closed, or closed. In other words, the terms also include "consisting essentially of" or "consisting of."

[0039] Annexin A2

[0040] Annexins are soluble proteins found in eukaryotes but absent in yeast and prokaryotes. Vertebrates have a total of 12 annexins, collectively known as annexins. Annexins bind to membranes, exposing negatively charged phosphatidylserine residues in a calcium-dependent manner. Annexin A2, a 39 kDa member of the annexin family, is expressed in most cells and tissues and binds to numerous ligands. Annexin A2 is a pleiotropic protein closely associated with transmembrane trafficking, cytoskeletal dynamics, extracellular receptor activity, and signal transduction. It promotes the invasion and migration of various tumor cells, the proliferation and survival of vascular endothelial cells, the migration of skin fibroblasts, and the repair of intestinal epithelial cells after injury. Currently, there is a lack of treatments for cartilage damage associated with osteoarthritis. Annexin A2, as a novel active ingredient, has enormous potential for promoting chondrocyte regeneration.

[0041] Annexin A2 that can be used in the present invention can be Annexin A2 derived from any organism, preferably from mammals (such as primates), more preferably from humans, monkeys, rats or mice. Annexin A2 of the present invention can also be a functional analog thereof, such as a protein having 50%, 60%, 70%, 75% or more, such as 80%, 85% or more, 90% or more, or even more preferably 95% or 99% or more identity with human Annexin A2; it should be understood that "Annexin A2" includes wild-type or mutant (including truncated) Annexin A2, as long as the mutant Annexin A2 retains or maintains the activity of wild-type Annexin A2, it can also be used in the present invention. In addition, Annexin A2 can also be a multimer, fusion protein, or chemically modified variant (such as PEG modification) of natural Annexin A2, provided that these variants possess the activity of wild-type Annexin A2.

[0042] The human annexin A2 sequence is shown in SEQ ID No: 1:

[0043] Osteoarthritis and its symptoms

[0044] Osteoarthritis (OA) is a cartilage degenerative disease that originates in the articular cartilage and gradually erodes the subchondral bone and surrounding tissues, leading to focal, erosive joint lesions, which in turn cause symptoms such as joint pain, joint stiffness, joint swelling, movement disorders, and deformities.

[0045] In the present invention, there are many factors that induce osteoarthritis, and the cause is not yet fully understood. It may be related to factors such as advanced age, obesity, drugs, and occupational overuse.

[0046] In the present invention, the occurrence site of osteoarthritis is not particularly limited. For example, it can be arthritis in weight-bearing joints and joints with high mobility, such as cervical vertebrae, lumbar vertebrae, knee joints, hip joints, etc.

[0047] The present invention is the first to discover that Annexin A2 can be used as an active ingredient to prevent or treat osteoarthritis.

[0048] The terms "prevent," "delay," "treat," "resist," or "improve" as used herein include delaying or alleviating one or more indicators or aspects associated with osteoarthritis to a certain extent, but do not necessarily require a 100% cure. In some embodiments, the active ingredient of the present invention or a composition comprising the same delays or alleviates one or more indicators associated with arthritis in a subject by, for example, at least about 5%, at least about 10%, at least about 15%, at least about 20%, or at least about 30%, compared to the absence of the active ingredient of the present invention.

[0049] Composition and administration

[0050] The composition of the present invention includes (but is not limited to): Annexin A2 and a pharmaceutically acceptable carrier.

[0051] The term "pharmaceutically or nutraceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gels suitable for human use, of sufficient purity, and with sufficiently low toxicity. "Compatibility" in this context refers to the ability of the components of the composition to blend with the active ingredient of the invention, and with each other, without significantly reducing the compound's efficacy. Such carriers include, but are not limited to, saline, buffered solutions, dextrose, water, glycerol, ethanol, and combinations thereof. The dosage form should be compatible with the route of administration.

[0052] Typically, the annexin A2 of the present invention can be prepared into pharmaceutical compositions in the form of tablets, capsules, powders, microgranules, solutions, lozenges, jellies, creams, spirits, suspensions, tinctures, poultices, liniments, lotions, and aerosols. Pharmaceutical compositions can be prepared using commonly known preparation techniques, and suitable pharmaceutical additives can be added to the composition.

[0053] The composition according to the present invention can be used for: parenteral, intravenous, intra-arterial, intraperitoneal, intramuscular or subcutaneous administration. Can be by the active ingredient of the required amount in a suitable solvent and various other ingredients (if necessary) as listed above merged, then by sterilization to make a sterile injectable solution. It is best to use the above composition in the form of a sterile aqueous solution, the sterile aqueous solution can contain other substances, such as enough salt or glucose, so that the solution and blood are isotonic. If necessary, the aqueous solution is appropriately buffered (preferably pH between 3-9). Standard pharmaceutical techniques well known to those skilled in the art can be utilized to easily prepare suitable pharmaceutical formulations under aseptic conditions. The dosage of the active ingredient is a therapeutically effective amount, for example, about 10 micrograms / kg body weight-about 50 mg / kg body weight every day.

[0054] The active ingredients of the present invention may be used alone or in combination with other drugs, such as anti-inflammatory and analgesic drugs.

[0055] The precise amount of active ingredient provided to an individual for a therapeutically effective amount will depend on the mode of administration, the type and severity of the disease and / or condition, and the characteristics of the individual, such as general health, age, sex, weight, and tolerance to the drug. One of ordinary skill in the art will be able to determine an appropriate dosage based on these and other factors. When administered in combination with other therapeutic agents, the "therapeutically effective amount" of any other therapeutic agent will depend on the type of drug used. Suitable dosages are known for approved therapeutic agents and can be adjusted by one of ordinary skill in the art based on the individual's condition, the type of condition being treated, and the amount of the compound of the present invention used below, for example, as reported in the literature and recommended in the Physician's Desk Reference (57th Edition, 2003). Preferably, the composition should be formulated so that an inhibitor dose of 0.01-100 mg / kg body weight / day can be administered to patients receiving these compositions. In certain embodiments, the compositions of the present invention provide a dosage of 0.01 mg to 50 mg. In other embodiments, a dosage of 0.1 mg to 25 mg or 5 mg to 40 mg is provided.

[0056] The present invention also provides a method for preventing or treating osteoarthritis, comprising administering Annexin A2 or the composition of the second aspect of the present invention to a subject in need thereof, thereby preventing or treating osteoarthritis.

[0057] Examples of subjects to whom the active ingredient of the present invention or a pharmaceutical composition comprising the same can be administered include mammals (eg, humans, mice, rats, hamsters, rabbits, cats, dogs, cows, sheep, monkeys, etc.).

[0058] The main advantages of the present invention include:

[0059] 1. The present invention is the first to discover the use of Annexin A2 in preventing or treating osteoarthritis.

[0060] 2. The present invention provides a new drug for preventing or treating osteoarthritis.

[0061] The present invention will be further described below in conjunction with specific implementation. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0062] Example 1

[0063] (1) Chondrocyte experiment

[0064] 1. Rat knee chondrocytes were cultured in vitro. Annexin A2 (SEQ ID No: 1) was added at varying concentrations during the culture process. Cell proliferation was detected using CCK8 assay after 72 hours of culture. The cell proliferation rate was calculated compared with that of the control group without drug addition.

[0065] 2. Rat knee chondrocytes were cultured in vitro. Sodium iodoacetate (final concentration 2 μM) was added during the culture process to create a cell injury model. Annexin A2 at different concentrations was also added. Cell proliferation was detected by CCK8 after 72 hours of culture. The cell proliferation rate was calculated compared with the control group without drug addition.

[0066] Experimental results

[0067] 1. As shown in Figure 1, the proliferation of rat chondrocytes was enhanced after treatment with AnnexinA2, with 0.25μg / ml having a better effect than 0.1μg / ml, showing a dose-dependent effect.

[0068] 2. As shown in Figure 2, the proliferation of rat chondrocytes was inhibited in the presence of iodoacetic acid, while the proliferation ability was protected in the presence of AnnexinA2.

[0069] Example 2

[0070] (2) Arthritis animal experiments

[0071] 1. SPF male Sprague-Dawley rats (8 weeks old), weighing 280-320 g, were acclimated for 2 weeks with free access to water and food in an animal room maintained at (25±2)°C, relative humidity 65%±3%, and a 12-h light / dark cycle.

[0072] 2. Animal grouping: control group, model group, and model + Annexin A2 low (0.1ug / ml) and high (1ug / ml) dose groups, with 5 animals in each group.

[0073] 3. Rats were anesthetized by inhalation of isoflurane (concentration: 2-3% for induction, 1.5-2% for maintenance). A rat knee osteoarthritis model was established by intra-articular injection of sodium iodoacetate. The injection dose was 1 mg of sodium iodoacetate (20 mg / ml) per joint in a 0.05 ml injection volume.

[0074] 4. Treatment: Seven days after the sodium iodoacetate injection, drug treatment was administered. Annexin A2 was injected intra-articularly at the designated dose (0.1 ml) once weekly for six weeks. The modeling group was injected with 0.1 ml of normal saline. Pain thresholds in the affected limb were monitored weekly during this period.

[0075] 6. Seven weeks after treatment, the animals were killed, and the knee joints of the rats were removed and fixed with 4% paraformaldehyde for 24 hours. The tissues were decalcified with EDTA decalcification solution for 4 weeks, dehydrated, paraffin-embedded, sectioned, and stained with HE.

[0076] Experimental results

[0077] 1. As shown in Figure 3, intra-articular injection of sodium iodoacetate can cause joint pain and shorten the pain threshold time. In the annexin A2 treatment group, the pain improved and gradually returned to normal with the increase in the number of treatments, and the higher the concentration, the better the effect; while the model group recovered slowly.

[0078] 2. As shown in Figure 4, histological analysis of the synovial tissue of the joints showed that there was a large amount of inflammatory cell infiltration in the synovium of the model group, while the inflammatory cell infiltration in the annexin A2 treatment group was not obvious, and the higher the concentration, the better the effect.

[0079] 3. As shown in Figure 5, histological analysis of articular cartilage showed degeneration and destruction of articular cartilage in the model group, while the articular cartilage in the annexin A2-treated group was intact, with no significant difference from the normal control group.

[0080] The above results show that Annexin A2 has the effect of reducing cartilage damage, promoting chondrocyte proliferation, reducing joint inflammation, alleviating joint pain and thus treating osteoarthritis.

[0081] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. Use of annexin A2 in the preparation of a pharmaceutical composition for preventing or treating osteoarthritis.

2. The use according to claim 1, characterized in that The osteoarthritis includes articular cartilage degeneration damage, and reactive hyperplasia of joint margin and / or subchondral bone caused by aging, obesity, strain, trauma and / or congenital abnormalities of joints.

3. The use according to claim 1, characterized in that The osteoarthritis is selected from the group consisting of cervical osteoarthritis, lumbar osteoarthritis, knee osteoarthritis, hip osteoarthritis, or a combination thereof.

4. The use according to claim 1, characterized in that The prevention or treatment of arthritis is carried out by one or more methods selected from the following groups: (a) Reduce cartilage damage; (b) Promote chondrocyte proliferation; (c) reduce inflammation in joints (including synovium); and (d) Relieve joint pain.

5. The use according to claim 1, characterized in that The pharmaceutical composition also includes a pharmaceutically acceptable carrier.

6. The use according to claim 1, characterized in that The dosage form of the pharmaceutical composition is an oral preparation, an external preparation or an injection preparation.

7. The use according to claim 1, characterized in that The composition is in the form of a solid dosage form, a semisolid dosage form, or a liquid dosage form, such as a tablet, a capsule, a microcapsule, a powder injection, an oral solution, an injection, a solution, a gel, an emulsion, an ointment, a cream, a paste, a powder, a patch, or a lozenge.

8. A pharmaceutical composition comprising (a) Annexin A2; and (b) a pharmaceutically acceptable carrier.

9. The use according to claim 1, characterized in that The injection preparation is an intravenous injection or an intramuscular injection.

10. The use according to claim 1, characterized in that The composition is in the form of a solid dosage form, a semisolid dosage form, or a liquid dosage form, such as a tablet, a capsule, a microcapsule, a powder injection, an oral solution, an injection, a solution, a gel, an emulsion, an ointment, a cream, a paste, a powder, a patch, or a lozenge.

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