CN118001373A - Medicine for treating osteoarthritis - Google Patents

Abstract

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

Description

A drug for treating osteoarthritis

Technical Field

The invention relates to the field of medicine, and in particular to a medicine for treating osteoarthritis.

Background technique

Osteoarthritis (OA) is the most common joint disease characterized by degeneration of articular cartilage. OA primarily affects the major weight-bearing joints, including the hip and knee. Clinical manifestations of OA include joint pain, swelling, limited mobility, and dysfunction. Changes in the tissues within OA joints are manifested as cartilage matrix destruction and reduction, osteophyte formation, subchondral bone sclerosis, and aseptic inflammation of the synovium. The occurrence of OA is influenced by multiple mechanisms, among which genetics, trauma, age, and obesity are the main factors. However, despite the increase in the incidence and severity of OA, there is currently no cure or effective disease-modifying therapy due to the poor understanding of the pathogenesis of the disease. Current pharmacological treatments for OA, such as corticosteroids, hyaluronic acid, and opioids, are mainly used to relieve symptoms that can neither prevent nor reverse cartilage degeneration. In addition, these drugs have side effects and toxicity. Due to irreversible disease progression, most patients eventually require joint replacement. These limitations require the development of new therapies that, in addition to 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 do not currently exist. Therefore, there is an urgent need to find effective drugs to delay the progression of OA and investigate its underlying molecular mechanisms.

Summary of the invention

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

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

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 joints.

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.

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

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

(a) Reduce cartilage damage;

(b) Promote chondrocyte proliferation;

(c) reduce inflammation in joints (including synovium);

(d) Relieve joint pain.

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

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

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

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 liquid, an injection, a solution, a gel, an emulsion, an ointment, a cream, a paste, a powder, a patch or a lozenge.

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

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

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

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

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

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 liquid, an injection, a solution, a gel, an emulsion, an ointment, a cream, a paste, a powder, a patch or a lozenge.

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

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

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.

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

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

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 specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 The promoting effect of Annexin A2 on chondrocyte proliferation.

Fig. 2 Protective effect of Annexin A2 on chondrocytes.

Figure 3 Annexin A2 treatment alleviates joint pain symptoms.

Figure 4 Annexin A2 inhibits inflammatory response in synovial membrane.

Figure 5 Annexin A2 protects cartilage from damage.

Detailed ways

After extensive and in-depth research, a large number of screenings and tests, the inventors provided for the first time the use of annexin A2 in preventing or treating osteoarthritis, and completed the present invention on this basis.

the term

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.

As used herein, the term "comprising" or "including (comprising)" may be open, semi-closed and closed. In other words, the term also includes "consisting essentially of" or "consisting of".

Annexin A2

Annexin is a soluble protein that exists in eukaryotes but not in yeast and prokaryotes. Vertebrates have a total of 12 annexins, named annexins. Annexins have the property of binding to membranes and exposing negatively charged phosphatidylserine in a calcium-dependent manner. Annexin A2 is a 39kDa member of the annexin family, expressed in most cells and tissues and bound to many ligands. Annexin A2 is a pleiotropic protein closely related to cell transmembrane transport, cytoskeleton 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 mucosal epithelial damage. At present, there is a lack of corresponding treatment for cartilage damage in osteoarthritis. Annexin A2, as a new active ingredient, plays a role in promoting chondrocyte regeneration and has great application potential.

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 polymer of natural Annexin A2, a fusion protein, or a chemically modified variant thereof (such as PEG modification), provided that these variants have the activity of wild-type Annexin A2.

The sequence of human annexin A2 is shown in SEQ ID No: 1:

1gctcagcatt tggggacgct ctcagctctc ggcgcacggc ccagcttcct tcaaaatgtc

61tactgttcac gaaatcctgt gcaagctcag cttggagggt gatcactcta cacccccaag

121tgcatatggg tctgtcaaag cctatactaa ctttgatgct gagcgggatg ctttgaacat

181tgaaacagcc atcaagacca aaggtgtgga tgaggtcacc attgtcaaca ttttgaccaa

241ccgcagcaat gcacagagac aggatattgc cttcgcctac cagagaagga ccaaaaagga

301acttgcatca gcactgaagt cagccttatc tggccacctg gagacggtga ttttgggcct

361attgaagaca cctgctcagt atgacgcttc tgagctaaaa gcttccatga aggggctggg

421aaccgacgag gactctctca ttgagatcat ctgctccaga accaaccagg agctgcagga

481aattaacaga gtctacaagg aaatgtacaa gactgatctg gagaaggaca ttatttcgga

541cacatctggt gacttccgca agctgatggt tgccctggca aagggtagaa gagcagagga

601tggctctgtc attgattatg aactgattga ccaagatgct cgggatctct atgacgctgg

661agtgaagagg aaaggaactg atgttcccaa gtggatcagc atcatgaccg agcggagcgt

721gccccacctc cagaaagtat ttgataggta caagagttac agcccttatg acatgttgga

781aagcatcagg aaagaggtta aaggagacct ggaaaatgct ttcctgaacc tggttcagtg

841cattcagaac aagcccctgt attttgctga tcggctgtat gactccatga agggcaaggg

901gacgcgagat aaggtcctga tcagaatcat ggtctcccgc agtgaagtgg acatgttgaa

961aattaggtct gaattcaaga gaaagtacgg caagtccctg tactattata tccagcaaga

1021cactaagggc gactaccaga aagcgctgct gtacctgtgt ggtggagatg actgaagccc

1081gacacggcct gagcgtccag aaatggtgct caccatgctt ccagctaaca ggtctagaaa

1141accagcttgc gaataacagt ccccgtggcc atccctgtga gggtgacgtt agcattaccc

1201ccaacctcat tttagttgcc taagcattgc ctggccttcc tgtctagtct ctcctgtaag

1261ccaaagaaat gaacattcca aggagttgga agtgaagtct atgatgtgaa acactttgcc

1321tcctgtgtac tgtgtcataa acagatgaat aaactgaatt tgtactttag aaacacgtac

1381tttgtggccc tgctttcaac tgaattgttt gaaaattaaa cgtgcttggg gttcagctgg

1441tgaggctgtc cctgtaggaa gaaagctctg ggactgagct gtacagtatg gttgccccta

1501tccaagtgtc gctatttaag ttaaatttaa atgaaataa ataaaataaa atca

Osteoarthritis and its symptoms

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.

In the present invention, there are many inducing factors for osteoarthritis, and the cause is still completely clear, which may be related to factors such as advanced age, obesity, drugs, occupational overuse, etc.

In the present invention, the occurrence site of osteoarthritis is not particularly limited, and may be, for example, weight-bearing joints and joints with a high amount of activity, such as arthritis of the cervical vertebrae, lumbar vertebrae, knee joints, hip joints, etc.

The present invention firstly finds that Annexin A2 can be used as an active ingredient to prevent or treat osteoarthritis.

The terms "prevention", "delay", "treatment", "resistance" or "improvement" described in the present invention include delaying or alleviating one or more indicators or aspects related to osteoarthritis to a certain extent, but do not necessarily require 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 related to arthritis of the subject of use 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.

Composition and administration

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

The term "pharmaceutically or health-care acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the active ingredients of the present invention and with each other without significantly reducing the efficacy of the compound. Such carriers include (but are not limited to): saline, buffer, glucose, water, glycerol, ethanol, and combinations thereof. The dosage form of the drug should match the mode of administration.

Typically, the annexin A2 of the present invention can be prepared into a pharmaceutical composition, such as a tablet, capsule, powder, microgranule, solution, lozenge, jelly, cream preparation, spirit, suspension, tincture, poultice, liniment, lotion, and aerosol. The pharmaceutical composition can be prepared by a commonly known preparation technique, and suitable pharmaceutical additives can be added to the drug.

Compositions according to the present invention can be used for: parenteral, intravenous, intra-arterial, intraperitoneal, intramuscular or subcutaneous administration. A sterile injectable solution can be prepared by merging the required amount of active ingredient in a suitable solvent with various other ingredients (if necessary) as listed above, and then sterilizing. It is best to use the above composition in the form of a sterile aqueous solution, which may contain other substances, such as sufficient salt or glucose, to make the solution isotonic with blood. If necessary, the aqueous solution is appropriately buffered (preferably pH between 3-9). Suitable pharmaceutical formulations can be easily prepared under aseptic conditions using standard pharmaceutical techniques well known to those skilled in the art. 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 per day.

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

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, body weight, and tolerance to the drug. One of ordinary skill in the art will be able to determine the 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 composition of the present invention provides a dosage of 0.01 mg to 50 mg. In other embodiments, a dosage of 0.1 mg-25 mg or 5 mg-40 mg is provided.

The present invention also provides a method for preventing or treating osteoarthritis, which comprises 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.

Examples of subjects to which 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.).

The main advantages of the present invention include:

1. The present invention is the first to discover the use of annexin A2 in preventing or treating osteoarthritis.

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

The present invention will be further described below in conjunction with specific implementation. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

Example 1

(1) Chondrocyte experiment

1. Rat knee chondrocytes were cultured in vitro, and different concentrations of AnnexinA2 (SEQ ID No: 1) were added during the culture process. After 72 hours of culture, CCK8 was used to detect cell proliferation, and the cell proliferation rate was calculated compared with the control group without drug addition.

2. Rat knee chondrocytes were cultured in vitro. Sodium iodoacetate (final concentration was 2 μM) was added during the culture process to create a cell injury model. Different concentrations of AnnexinA2 were added at the same time. After 72 hours of culture, cell proliferation was detected by CCK8. The cell proliferation rate was calculated compared with the control group without drug addition.

Experimental Results

1. As shown in Figure 1, the proliferation ability of rat chondrocytes was enhanced after treatment with AnnexinA2, and the concentration of 0.25μg/ml had a better effect than 0.1μg/ml, showing a dose-dependent effect.

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

Example 2

(2) Arthritis animal experiments

1. SPF grade SD male rats (8 weeks old), weighing 280-320 g, with free access to water and food, and the room temperature of the animal room was (25±2)℃, relative humidity was 65%±3%, and the light/dark period was 12h per day. The animals were adaptively raised for 2 weeks.

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.

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

4. Treatment: Drug treatment was given 7 days after sodium iodoacetate injection. Annexin A2 was injected into the joint cavity at a specified dose with an injection volume of 0.1 ml, once a week for six consecutive weeks. The modeling group was injected with 0.1 ml of normal saline. The pain threshold of the affected limb was tested weekly during this period.

5. Seventh week after treatment, the animals were killed, and the knee joints of the rats were fixed with 4% paraformaldehyde for 24 h, decalcified with EDTA decalcification solution for 4 weeks, and the tissues were dehydrated, embedded in paraffin, sliced, and stained with HE.

Experimental Results

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.

2. As shown in Figure 4, histological analysis of synovial tissue showed that there were a large number of inflammatory cells infiltrating the synovium in the model group, while there was no obvious inflammatory cell infiltration in the annexin A2 treatment group, and the higher the concentration, the better the effect.

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 treatment group was intact and had no significant difference from the normal control group.

The above results prove that annexin A2 has the effect of reducing cartilage damage, promoting chondrocyte proliferation, reducing joint inflammation, alleviating joint pain and thus treating osteoarthritis.

All documents mentioned in the present invention are cited as references in this application, just as each document is cited as references individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims (10)

1. Use of annexin A2 for the preparation of a pharmaceutical composition for the prevention or treatment of osteoarthritis.

2. The use according to claim 1, wherein the osteoarthritis comprises degenerative damage of articular cartilage, joint edge and/or subchondral bone reactive hyperplasia caused by aging, obesity, strain, trauma and/or congenital abnormalities of the joint.

3. The use according to claim 1, wherein 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, wherein the prevention or treatment of arthritis is carried out by one or more means selected from the group consisting of:

(a) Reducing cartilage damage;

(b) Promoting chondrocyte proliferation;

(c) Reducing joint (including synovial) inflammatory responses; and

(D) Relieving joint pain.

5. The use of claim 1, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

6. The use according to claim 1, wherein the pharmaceutical composition is in the form of an oral preparation, an external preparation or an injectable preparation.

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

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

9. The use according to claim 1, wherein the injectable formulation is an intravenous or intramuscular injection.

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