A composition for treating joint diseases or connective tissue diseases, comprising dextran or poloxamer

Abstract

The present invention relates to a composition for treating joint diseases or connective tissue diseases, which contains dextran, poloxamer or a mixture thereof, or dextran, a composition for cartilage regeneration, which contains poloxamer or a mixture thereof. The composition of the present invention can stay in the damaged part of a joint or connective tissue for a long time due to a buffering effect, a coating effect or an anti-inflammatory effect, can relieve shock and specifically wrap the damaged part in a damaged part-specific manner, or reduce inflammation of an adhered part, and thus can be effectively used for treating joint diseases, treating connective tissue diseases or cartilage regeneration.

Description

Technical Field

[0001] The present invention relates to a composition for treating joint diseases or connective tissue diseases, a composition for cartilage regeneration, or a composition for treating inflammatory diseases, which comprises dextran, poloxamer or a mixture thereof. Background Art

[0002] A joint is a part where bones meet, and is composed of cartilage, joint capsule, synovium, ligaments, tendons, muscles, etc., so that bones can move smoothly and play a role in absorbing the impact generated by movement. Arthritis refers to joint dysfunction caused by various reasons, such as accompanying inflammation, etc. Connective tissue is also called connective tissue, and is a tissue that forms organs by combining tissues in animals. For example, cartilage, intervertebral discs, tendons, ligaments, bones, skin, adipose tissue, blood vessels, intestinal tissue, etc. can be cited.

[0003] Drugs for treating arthritis can be roughly classified according to the main mechanisms of action such as reducing inflammation, delaying the progression of the disease, and reducing the concentration of metabolic products such as uric acid. Many drugs for treating arthritis have the effect of reducing inflammation. Inflammation is a pathological process that causes pain, edema, fever, redness, stiffness, etc. Drugs that rapidly relieve inflammation include non-steroidal anti-inflammatory agents including aspirin and steroidal anti-inflammatory agents including cortisone. However, the main effects of these anti-inflammatory and anti-inflammatory agents are to relieve pain, and various complications will occur when taking the drugs for a long time. In particular, steroidal anti-inflammatory agents have nothing to do with the treatment of the cause of the disease, and due to their strong temporary anti-inflammatory effect, there is a possibility of inducing excessive use of joints by simply temporarily relieving pain, which will become a factor in damaging joints and exacerbating disability. Therefore, they need to be used with caution.

[0004] Therefore, the effectiveness of existing treatment methods for joint damage such as arthritis is limited, accompanied by obvious toxic and side effects. Since they cannot be continuously used for a long time, their effectiveness is limited. Therefore, there is an urgent need for a new treatment method or therapeutic agent that overcomes the disadvantages of existing treatment methods.

[0005] On the other hand, dextran is one of polysaccharides, which refers to a polymer of D-glucose and has a structure similar to starch or glycogen. D-glucose is connected in a linear shape by α-1,6 bonds and branched at multiple points by α-1,4 or α-1,3 bonds. It is known that commonly used high molecular weight dextran often causes allergic reactions in the body. Dextran has been used as a plasma volume expander, a blood anticoagulant (anti-adhesion agent), a filler using cross-linked dextran, etc. However, it has not been reported that it is used as a therapeutic agent for joint diseases or connective tissue diseases. Poloxamer refers to a non-ionic surfactant, and in addition to the surfactant, it is known to be used as an emulsifier, a stabilizer, a co-solvent, etc.

[0006] At present, due to the limitations of existing treatments for joint diseases or connective tissue diseases, namely, limited effectiveness, obvious toxic side effects, and lack of persistence, new treatments or therapeutic agents are needed. There is no research on the treatment of joint diseases or connective tissue diseases using dextran or poloxamer, especially low-molecular-weight dextran or poloxamer, which does not cause allergic reactions, especially when administered in vivo. Summary of the Invention

[0007] For this reason, the present inventors conducted research on the treatment of joints or connective tissues, and as a result, confirmed that dextran, poloxamer, or a mixture thereof does not cause allergic reactions, and protects joints and connective tissues through the buffering effect, preservation effect, or anti-inflammatory effect on the damaged parts of joints and connective tissues, thereby completing the present invention.

[0008] An object of the present invention is to provide a pharmaceutical composition for treating joint diseases or connective tissue diseases, a composition for regenerating cartilage, or a pharmaceutical composition for treating inflammatory diseases, which contains dextran, poloxamer, or a mixture thereof.

[0009] To achieve the above object, the present invention provides a pharmaceutical composition for treating joint diseases or connective tissue diseases, which contains dextran, poloxamer, or a mixture thereof.

[0010] Furthermore, the present invention provides a pharmaceutical composition for preventing or treating joint diseases or connective tissue diseases, which contains a mixture of two selected from the group consisting of dextran 1, dextran 5, and poloxamer 188.

[0011] Furthermore, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases, which contains dextran, poloxamer, or a mixture thereof as an active ingredient.

[0012] Furthermore, the present invention provides a composition for regenerating cartilage, which contains dextran, poloxamer, or a mixture thereof as an active ingredient.

[0013] Furthermore, the present invention provides a composition for regenerating cartilage, which contains a mixture of two selected from the group consisting of dextran 1, dextran 5, and poloxamer 188.

[0014] The composition of the present invention can stay in the damaged parts of joints or connective tissues for a long time due to the buffering effect, coating effect, or anti-inflammatory effect, can relieve shock and specifically wrap the damaged parts in a damaged part-specific manner, or reduce the inflammation of the adhered parts, and thus can be effectively used for the treatment of joint diseases, connective tissue diseases, or cartilage regeneration. Brief Description of the Drawings

[0015] Figure 1 Figure showing the results of confirming the cell proliferation ability of chondrocytes after treating normal chondrocytes with dextran, poloxamer, or their mixture for 24 hours (* / ** / *** indicate significance of p < 0.05 / p < 0.01 / p < 0.001 relative to the negative control group (nc), respectively).

[0016] Figure 2 Figure showing the results of confirming the cell proliferation ability of chondrocytes after treating normal chondrocytes with dextran, poloxamer, or their mixture for 48 hours (* / ** / *** indicate significance of p < 0.05 / p < 0.01 / p < 0.001 relative to the negative control group (nc), respectively).

[0017] Figure 3 Figure showing the results of confirming the cell proliferation ability of chondrocytes after treating chondrocytes in an in vitro osteoarthritis model treated with recombinant human interleukin-1α (rhIL-1α) with dextran, poloxamer, or their mixture for 24 hours (* / ** / *** indicate significance of p < 0.05 / p < 0.01 / p < 0.001 relative to the inflammation-induced positive control group (pc), respectively).

[0018] Figure 4 Figure showing the results of confirming the cell proliferation ability of chondrocytes after treating chondrocytes in an in vitro osteoarthritis model treated with recombinant human interleukin-1α with dextran, poloxamer, or their mixture for 48 hours (* / ** / *** indicate significance of p < 0.05 / p < 0.01 / p < 0.001 relative to the inflammation-induced positive control group (pc), respectively).

[0019] Figure 5 Figure showing the results of confirming the interleukin-10 (IL-10) / interleukin-6 (IL-6) gene expression ratio after treating chondrocytes in an in vitro osteoarthritis model treated with recombinant human interleukin-1α with dextran, poloxamer, or their mixture (* / ** / *** indicate significance of p < 0.05 / p < 0.01 / p < 0.001 relative to the inflammation-induced positive control group (pc), respectively).

[0020] Figure 6 Figure showing the results of confirming the expression level of matrix metalloproteinase-3 (MMP-3) gene after treating chondrocytes in an in vitro osteoarthritis model treated with recombinant human interleukin-1α with dextran, poloxamer, or their mixture (* / ** / *** indicate significance of p < 0.05 / p < 0.01 / p < 0.001 relative to the inflammation-induced positive control group (pc), respectively).

[0021] Figure 7 A graph showing the results of confirming the expression level of matrix metalloproteinase-13 (MMP-13) gene after treating chondrocytes in an in vitro osteoarthritis model treated with recombinant human interleukin-1α with dextran, poloxamer, or a mixture thereof (* / ** / *** indicate significance of p < 0.05 / p < 0.01 / p < 0.001 relative to the inflammation-induced positive control group (pc), respectively).

[0022] Figure 8 A graph showing the results of confirming the production amount of type II collagen after treating chondrocytes in an in vitro osteoarthritis model treated with recombinant human interleukin-1α with dextran, poloxamer, or a mixture thereof (* / ** / *** indicate significance of p < 0.05 / p < 0.01 / p < 0.001 relative to the inflammation-induced positive control group (pc), respectively).

[0023] Figure 9 A graph showing the results of confirming the production amount of proteoglycan after treating chondrocytes in an in vitro osteoarthritis model based on recombinant human interleukin-1α with dextran, poloxamer, or a mixture thereof (* / ** / *** indicate significance of p < 0.05 / p < 0.01 / p < 0.001 relative to the inflammation-induced positive control group (pc), respectively). Detailed Description of the Invention

[0024] Best Mode for Carrying Out the Invention

[0025] Hereinafter, the present invention will be described in detail.

[0026] The present invention provides a pharmaceutical composition for treating joint diseases or connective tissue diseases, comprising dextran, poloxamer, or a mixture thereof. Further, the present invention provides a method for preventing or treating joint diseases or connective tissue diseases, comprising the step of administering dextran, poloxamer, or a mixture thereof to a subject in need thereof.

[0027] In the present invention, "dextran" refers to a polymer of D-glucose as one of the polysaccharides. Dextran has a structure similar to starch or glycogen, and D-glucose is connected in a straight-chain shape by α-1,6 bonds and branched at multiple points by α-1,4 or α-1,3 bonds. Dextran can be used as a blood volume expander and a blood anticoagulant, and depending on the purpose, it can be used by changing the molecular weight of dextran. In the present invention, the above dextran may have a molecular weight of 500 Da to 10,000 Da, preferably 500 Da to 8,000 Da, and more preferably may have a molecular weight of 1,000 Da to 5,000 Da. Dextran is named "dextran 1" when its molecular weight is 1,000 Da, "dextran 5" when its molecular weight is 5,000 Da, "dextran 10" when its molecular weight is 10,000 Da, etc.

[0028] Generally, it is known that when dextran with a molecular weight of 8,000 Da or more is administered to the human body, there is a risk of causing an allergic reaction. In particular, as the above allergic reaction, anaphylaxis, which is a rapid systemic reaction caused by an antigen-antibody immune reaction, may occur, but this is very dangerous. However, for the low-molecular-weight dextran according to the present invention, if dextran with a molecular weight of 1,000 Da to 5,000 Da is preferably used, it has the advantage of minimizing the allergic reaction, and compared with known substances (e.g., hyaluronic acid), the decomposition rate in the body is very slow. Therefore, it has the advantage of being able to maintain the therapeutic effect of joint diseases or connective tissue diseases for a long time.

[0029] Moreover, compared with dextran with a molecular weight of 8,000 Da or more, the low-molecular-weight dextran according to the present invention has the following characteristics: not only does it not cause an allergic reaction, but also the onset of the rental time is very fast and it can be distributed within a wide range.

[0030] In the present invention, examples using dextran 1 and dextran 5 are provided as preferred examples. They can be used in combination with dextran 1 and dextran 5, and can also be used in combination with poloxamer 188.

[0031] In the present invention, two selected from the group consisting of dextran 1, dextran 5, and poloxamer 188 can be mixed at a volume ratio of 1:1.

[0032] In the present invention, the concentration of the above dextran is expressed as a percentage of the mass (g) concentration (w / v) in 100 cc of solvent. For example, when dextran 1 is included as a single active ingredient in the composition, dextran 1 with a concentration of 2 (w / v)% to 40 (w / v)% can be included, and when dextran 5 is included as a single active ingredient in the composition, dextran 5 with a concentration of 2 to 30 (w / v)% can be included.

[0033] Moreover, when dextran is included in the composition as a mixture of dextran 1 or dextran 5, or a mixture of dextran and poloxamer, the concentration of dextran may vary depending on the mixing target. For example, when a mixture of dextran 5 and dextran 1 is included in the composition, the concentration of the above dextran 5 is 2 (w / v)% to 40 (w / v)%, the concentration of the above dextran 1 is 2.5 (w / v)% to 40 (w / v)%, or the concentration of dextran 5 is 4 (w / v)% to 40 (w / v)%, and the concentration of dextran 1 can be 5 (w / v)% to 40 (w / v)%.

[0034] When dextran is present in the composition at a high concentration, due to the characteristics of the water containing polysaccharides, there is a disadvantage that the viscosity increases excessively and it is difficult to form a shape. Therefore, dextran is usually used at a concentration of 5% or less. However, the dextran in the composition of the present invention contains at least 240 (w / v)% to at most 40 (w / v)% concentration according to the molecular weight, so the therapeutic effect of joint diseases or connective tissue diseases can be maximized. In order to improve or stabilize the solubility of this dextran, the composition of the present invention may further contain additives, and the above additives may include sugar alcohols, monosaccharides or divalent cations containing CaCl 2 2.

[0035] In the present invention, "poloxamer" is a non-ionic triblock copolymer in which hydrophobic propylene oxide is centered and hydrophilic ethylene oxide is bonded at both ends. Due to its temperature-sensitive characteristics, it can be transformed into sol and gel according to the concentration and temperature, and the properties of poloxamer vary according to the ratio of polyoxypropylene and polyoxyethylene. In addition to surfactants, poloxamer can also be used as an emulsifier, stabilizer, co-solvent, etc.

[0036] In the present invention, the above poloxamer may have an average molecular weight of 100 Da to 20,000 Da.

[0037] The above-mentioned poloxamer is optionally selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 105 benzoate, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 182 dibenzoate, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403 and poloxamer 407. Preferably, it includes poloxamer 188, but is not limited thereto.

[0038] In the present invention, the concentration of the above-mentioned poloxamer refers to the percentage of the mass (g) concentration (w / v) in 100 cc of solvent. It is characterized in that it can be included in the composition at a concentration of 1 (w / v)% to 50 (w / v)%, or 2 (w / v)% to 30 (w / v)%. For example, when poloxamer 188 is included as a single active ingredient in the composition, it can be included in the composition at a concentration of 1 (w / v)% to 15 (w / v)% or 1 (w / v)% to 10 (w / v)%.

[0039] In order to increase or stabilize the solubility of such poloxamer, the composition of the present invention may further contain additives such as sugar alcohols, monosaccharides, divalent cations, etc., and sterilized water or physiological saline may also be included in the composition without additional additives.

[0040] In the present invention, the above-mentioned pharmaceutical composition may contain a mixture of dextran and poloxamer. In particular, when poloxamer 188 is used in combination with dextran, it can be used in combination with dextran 1 or dextran 5. For example, when combined with dextran 5, the concentration of the above-mentioned dextran 5 is 2 (w / v)% to 40 (w / v)%, and the concentration of the above-mentioned poloxamer 188 can be 1 (w / v)% to 20 (w / v)%, or the concentration of dextran 5 is 4 (w / v)% to 40 (w / v)%, and the concentration of poloxamer 188 can be 2 (w / v)% to 10 (w / v)%.

[0041] Also, for example, when mixed with dextran 1, the concentration of the above dextran 1 is 2.5 (w / v)% to 40 (w / v)%, the concentration of the above poloxamer can be 1 (w / v)% to 5 (w / v)%, or the concentration of dextran 1 is 45 (w / v)% to 55 (w / v)%, and the concentration of poloxamer 188 can be 1 (w / v)% to 2 (w / v)%.

[0042] When poloxamer 188 is contained in the composition at a high concentration, for example, greater than 25 (w / v)%, it may be toxic and can be used by appropriately adjusting the concentration when mixed with dextran.

[0043] As an example, for instance, dextran:poloxamer can be contained in the pharmaceutical composition of the present invention at (w / v)% of 5:0.05, 5:0.5, 5:5, 5:10, 5:20, 5:30, 5:40, 5:50, 5:100, 5:150, 5:200, 10:0.1, 10:1, 10:10, 10:20, 10:30, 10:40, 10:50, 10:100, 10:150, 10:300, 10:400, 50:0.5, 50:1, 50:5, 50:10, 50:20, 50:30, 50:40, 50:50. For example, in the case of mixing 1 (w / v)% of poloxamer 188 and 40 (w / v)% of dextran 1, it reflects a ratio of 1:40, and in the case of mixing 20 (w / v)% of poloxamer 188 and 2 (w / v)% of dextran 5, it reflects a ratio of 1:0.1, and can be contained in the pharmaceutical composition of the present invention at 1:0.1 to 1:40. Among them, the average molecular weight of the above dextran can be 1000 Da or 5000 Da, and poloxamer 188 with an average molecular weight of 8500 Da of the above poloxamer can be contained. Also, as an example of the present invention, when containing a mixture of two selected from the group consisting of dextran 1, dextran 5, and poloxamer 188, each component is mixed at a volume ratio of 1:1 and can be contained in the composition.

[0044] Moreover, as an embodiment of the present invention, the pharmaceutical composition of the present invention comprises a mixture of dextran 1 and dextran 5. For example, dextran 1:dextran 5 can be included in the pharmaceutical composition in a (w / v)% ratio of 5:0.05, 5:0.5, 5:5, 5:10, 5:20, 5:30, 5:40, 5:50, 5:100, 10:0.1, 10:1, 10:10, 10:20, 10:30, 10:40, 10:50, 10:100, 10:150, 10:200, 50:0.5, 50:1, 50:5, 50:10, 50:20, 50:30, 50:40, 50:50, 50:1000. For example, in the case of mixing 2.5 (w / v)% of dextran 1 and 40 (w / v)% of dextran 5 in an exemplary combination, it reflects a ratio of 1:16, and in the case of mixing 40 (w / v)% of dextran 1 and 2 (w / v)% of dextran 5, it reflects a ratio of 1:0.05, and it can be included in the pharmaceutical composition in a ratio of 1:0.05 to 1:16.

[0045] In the case where the composition of the present invention comprises a mixture of dextran and poloxamer, the hydrophobic component of poloxamer can increase the solubility of dextran or the combined drug, and polyethylene glycol (PEG), which is the hydrophilic component of poloxamer, can preserve tissues by preserving cell membranes and preventing adhesion. However, poloxamer is easily diluted by body fluids and has the disadvantage of being easily absorbed by body fluids, so its in vivo stability can be increased by mixing with dextran. Thus, in the case of a mixture of dextran and poloxamer, compared with the case of containing dextran alone or poloxamer alone, it can wrap the damaged joint or connective tissue site by maintaining viscosity for a long time, and reduce the inflammatory phenomenon caused by friction by preventing pathological adhesion of tissues, thereby smoothly providing nutrients and having the effect of promoting tissue regeneration. The above effects can be induced by the cushion effect, coating effect, or anti-inflammatory effect of the dextran and poloxamer mixture.

[0046] In the present invention, the "cushion effect" refers to the effect of acting like a cushion to relieve the applied impact by reducing the friction at the administration site such as a joint, and the "coating effect" refers to the effect of coating by quickly covering dry sites such as damaged cartilage tissue in a joint, resulting in the effect of comfortably moving the knee joint. The inflammatory response at the damaged joint site can be reduced through these effects.

[0047] Therefore, the composition of the present invention that induces buffering effects and coating effects or anti-inflammatory effects is suitable for improving regenerated or damaged tissues, and thus can be effectively used for the treatment of joint diseases or connective tissue diseases. The above-described effects of the composition of the present invention are similar to those of stem cell therapeutics currently used for the treatment of joint diseases, and it is competitive compared to stem cell therapeutics that require high technical skills and high costs.

[0048] On the other hand, a pharmaceutical composition for preventing or treating joint diseases or connective tissue diseases, which contains the dextran, poloxamer or a mixture thereof of the present invention, may further contain stem cells in order to enhance the preventive or therapeutic effects of joint diseases or connective tissue diseases required by the present invention. Stem cells that can be included in the present invention include embryonic stem cells or adult stem cells, and the above adult stem cells may be mesenchymal stem cells, human tissue-derived mesenchymal stromal cells, human tissue-derived mesenchymal stem cells, pluripotent stem cells or amniotic epithelial cells. The above stem cells may be stem cells derived from umbilical cord, umbilical cord blood, bone marrow, fat (or adipose tissue cells), muscle, nerve, skin, amnion and placenta (or placental tissue cells), urine, etc., but are not limited thereto. The above stem cells may be stem cells or their concentrates, stem cell culture solutions or their concentrates, culture secretions of stem cells or their concentrates or complexes thereof. The dextran or poloxamer in the above composition can play a role in specifically transporting stem cells to damaged lesion sites of joints or connective tissues, and has the effect of enhancing the regeneration of damaged tissues by attaching to the lesion sites where stem cells are transported.

[0049] In the present invention, "prevention" refers to all actions of inhibiting joint diseases or connective tissue diseases or delaying the onset by administering the composition.

[0050] In the present invention, "treatment" refers to all actions of improving or making favorable the symptoms of joint diseases or connective tissue diseases by administering the composition.

[0051] The pharmaceutical composition of the present invention may further contain pharmaceutically acceptable additives, and according to the conventional methods in the pharmaceutical field, it can be formulated into a unit dosage form preparation suitable for administration in a patient's body. And, it can be formulated by combining with pharmaceutically acceptable carriers or media, such as sterile water, physiological saline, vegetable oil, emulsifier, suspending agent, surfactant, stabilizer, excipient, vehicle, preservative or binder, etc., and mixing in a pharmaceutically acceptable unit dosage form.

[0052] The above pharmaceutical composition can be in a solution state or a powder state. In the case where the above pharmaceutical composition is in a powder state, it can be used by dissolving it in a solvent before administration, but it is not limited thereto. Considering various situations that may occur during the preparation of the drug, it can be prepared in the most suitable manner.

[0053] There is no particular limitation on the administration form of the above pharmaceutical composition. However, it can be administered through conventional administration routes such as oral administration, injection, and administration by means of infusion.

[0054] In the case of oral administration, it can also be used as a composition having the above composition, or used as a preparation such as tablets, pills, capsules, gels, syrups, etc. together with pharmaceutically acceptable carriers and excipients. However, since it takes time for solid preparations such as tablets or powders to be absorbed, oral administration by a liquid preparation is preferred. In this case, preferably, it can be administered as an aqueous solution together with appropriate additives such as salts such as sodium chloride, buffers, chelating agents, etc.

[0055] Moreover, the drug can be administered through a targeted drug delivery system such as liposomes coated with specific antibodies targeting joint diseases or connective tissues. Liposomes can be selectively taken from diseased tissues.

[0056] In the case of administering the pharmaceutical composition of the present invention as a locally administered preparation, appropriate buffers, isotonic agents, etc. are added and it is dissolved in sterilized distilled water and directly injected into the joint cavity, connective tissue, vein, subcutaneous, intradermal, intra-articular or muscle of an individual, or applied to the skin or can be in the form of a patch.

[0057] The above individual is one selected from mammals including humans, dogs, cats, pigs, horses, sheep, mice, and monkeys. Preferably, it can be a human.

[0058] In the present invention, the term "intra-articular" refers to percutaneous injection of the pharmaceutical composition of the present invention into a joint.

[0059] In the present invention, the term "local administration" refers to percutaneous injection into or near an inflamed joint. Therefore, local administration injection is related to the epidermis, dermis, muscle, or any organ of the body.

[0060] The main advantages of local administration are that the analgesic effect is selectively restricted to the damaged area. Furthermore, local administration allows a high local concentration level with little or no systemic release.

[0061] The above pharmaceutical composition may also contain stabilizers, lubricants, buffers, isotonicity regulators, anesthetics, or antibacterial agents, etc.

[0062] In addition, the above pharmaceutical composition may further contain anti-inflammatory agents widely used in the art. The above anti-inflammatory agents can be non-steroidal, steroidal or their complexes. Non-limiting examples of non-steroidal anti-inflammatory agents include: oxicams, such as piroxicam, isoxicam, tenoxicam, sudoxicam; salicylates, such as aspirin, salsalate, benorilate, choline magnesium trisalicylate, safapryn, solprin, diflunisal and fenbufen; acetic acid derivatives, such as diclofenac, fenclofenac, indomethacin, sulindac, tolmetin, isoxepac, furclofenac, tiopinac, zidometacin, acemetacin, fentiazac, zomepirac, clopirac, oxepinac, biphenylacetic acid and ketorolac; fenamates; fenamates, such as mefenamic acid, meclofenamic acid, flufenamic acid, niflumic acid and tolfenamic acid; propionic acid derivatives, such as ibuprofen, naproxen, benoxaprofen, flurbiprofen, ketoprofen, fenoprofen, fenbufen, indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen, miroprofen, tiaprofenic acid, sulprofen, alminoprofen and tiaprofenic acid; pyrazoles, such as phenylbutazone, oxyphenbutazone, febuprazone, azapropazone and trimethazone. Extracts of these non-steroidal anti-inflammatory agents can also be used.

[0063] Non-limiting examples of the above steroidal anti-inflammatory agents include corticosteroids, such as hydrocortisone, hydroxy-triamcinolone, α-methyl dexamethasone, dexamethasone phosphate, beclomethasone dipropionate, clobetasol propionate, desonide, desoxymethasone, desoxycorticosterone acetate, dexamethasone, dichlorisone, diflorasone diacetate, diflucortolone valerate, fluocinolone acetonide, flunisolide, fludrocortisone, flumethasone pivalate, fluocinonide, fluocinonide acetate, fluocortin butyl, flucortolone, fluprednidene acetate, fluocinolone acetonide, halcinonide, hydrocortisone acetate, hydrocortisone butyrate, methylprednisolone, triamcinolone acetonide, cortisone, cortodoxone, flucetonide, fludrocortisone, diflorasone diacetate, fluocinolone acetonide, fludrocortisone, fludrocortisone acetate, fluprednisolone, fluradrenolone acetonide, medrysone, amcinafal, amcinafide, betamethasone and its balanced esters, chloroprednisone, chloroprednisone acetate, clocortolone, ciclesonide, dichlorisone, difluprednate, flunisolide, flunisolide acetate, fluorometholone, fluperolone, fluprednisolone, hydrocortisone valerate, hydrocortisone cypionate, hydrocortamate, methylprednisolone acetate, paramethasone, prednisolone, prednisone, beclomethasone dipropionate, triamcinolone and its extracts.

[0064] The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease, and the effective dosage level can depend on factors including the severity of the disease, the age, weight, health, gender of the patient, the sensitivity of the patient to the drug, the route of administration and the excretion rate, the treatment time, the drugs used in combination with or simultaneously with the composition of the present invention, as well as other factors well known in the medical field. The dosage of dextran, poloxamer or a mixture thereof varies within a wide range and depends on the individual requirements of each particular case.

[0065] The pharmaceutical composition of the present invention can be co-administered with known medicaments for treating joint diseases or connective tissue diseases, and can be administered simultaneously with or sequentially to the above-mentioned therapeutic agents.

[0066] The pharmaceutical composition of the present invention can be divided into one or several doses, with a preferred dosing interval. For example, it can be administered every 1 week.

[0067] According to the present invention, a pharmaceutical composition comprising dextran, poloxamer or a mixture thereof can induce alleviation of long-term pain in an individual suffering from a joint disease or a connective tissue disease. In the case of using the pharmaceutical composition according to the present invention, major drug side effects are avoided, and it is possible to treat and / or prevent acute and chronic joint diseases or connective tissue diseases without using overly invasive techniques.

[0068] In the present invention, "connective tissue" is also referred to as "connective tissue", which refers to the tissue that is widely distributed in animal tissues and is used for binding, protecting, filling cells, viscera, and organs. It is divided into fibrous connective tissue, glial tissue, and reticular tissue. However, fibrous connective tissue is in the majority. For example, the connective tissue of the present invention can include, but is not limited to, cartilage, intervertebral discs, tendons, ligaments, bones, skin, blood vessels, intestinal tissues, etc.

[0069] In the present invention, the causes of "joint diseases" or "connective tissue diseases" are roughly classified into traumatic, infectious, inflammatory, and degenerative. The composition or therapeutic agent according to the present invention can be used for joint diseases and connective tissue diseases caused by various reasons that damage joints. However, the joint diseases for which the composition of the present invention is expected to be effectively used include osteoarthritis, preferably, recurrent friction and resulting degenerative osteoarthritis, ankylosing spondylitis, psoriatic arthritis, traumatic arthritis, rheumatoid arthritis, patellofemoral pain syndrome, chronic inflammation or arthrosis. And, the connective tissue diseases for which the composition of the present invention is expected to be effectively used include connective tissue diseases in cases where degenerative changes caused by physical damage or recurrent friction to cartilage, intervertebral discs, tendons, ligaments, bones, skin, blood vessels, intestinal tissues, etc. result in loss of collagen or moisture. In particular, the composition of the present invention has excellent effects of promoting anti-inflammatory effects and synthesis of collagen, and increasing the production amount of proteoglycan. Therefore, based on these effects, it can be effectively used in the prevention or treatment of connective tissue diseases, especially connective tissue diseases related to inflammation, preferably, one or more connective tissue diseases selected from the group consisting of inflammatory osteoarticular diseases, inflammatory dermatitis, inflammatory eye diseases, inflammatory myositis, inflammatory gastrointestinal diseases, chondropathy, vasculitis, and sprains.

[0070] In the present invention, the following preferred examples are provided, so as to achieve the effects of not showing cytotoxicity and effectively showing anti-inflammatory effects, inhibiting the gene expression of matrix metalloproteinase-3 and matrix metalloproteinase-13, which are proteins that destroy the cartilage matrix, promoting the synthesis of type II collagen as a cartilage protection effect, and increasing the production amount of proteoglycan.

[0071] In the present invention, two selected from the group consisting of dextran 1, dextran 5, and poloxamer 188 can be mixed at a volume ratio of 1:1.

[0072] Furthermore, the present invention relates to a pharmaceutical composition for preventing or treating joint diseases or connective tissue diseases, which contains a mixture of two selected from the group consisting of dextran 1, dextran 5, and poloxamer 188.

[0073] The above dextran is dextran 1, which is included as a single active ingredient in the composition, and the concentration of dextran 1 can be 2 (w / v)% to 40 (w / v)%.

[0074] The above dextran is dextran 5, which is included as a single active ingredient in the composition, and the concentration of dextran 5 can be 2 (w / v)% to 30 (w / v)%.

[0075] Furthermore, the above poloxamer is included as a single active ingredient in the composition, and the concentration of poloxamer can be 1 (w / v)% to 15 (w / v)%.

[0076] In one example of the present invention, the above mixture is a mixture of dextran 5 and poloxamer, the concentration of the above dextran 5 is 2 (w / v)% to 40 (w / v)%, and the concentration of the above poloxamer can be 1 (w / v)% to 20 (w / v)%.

[0077] Moreover, the above mixture is a mixture of dextran 5 and dextran 1, the concentration of the above dextran 5 is 2 (w / v)% to 40 (w / v)%, and the concentration of the above dextran 1 can be 2.5 (w / v)% to 40 (w / v)%.

[0078] Moreover, the above mixture is a mixture of dextran 1 and poloxamer, the concentration of the above dextran 1 is 2.5 (w / v)% to 40 (w / v)%, and the concentration of the above poloxamer can be 1 (w / v)% to 5 (w / v)%.

[0079] Moreover, the above mixture is a mixture of dextran 1 and poloxamer, the concentration of the above dextran 1 is 45 (w / v)% to 55 (w / v)%, and the concentration of the above poloxamer can be 1 (w / v)% to 2 (w / v)%.

[0080] Moreover, one example of the present invention relates to a composition based on an anti-inflammatory effect, a pharmaceutical composition for preventing or treating inflammatory diseases, and a method for preventing or treating inflammatory diseases using the same.

[0081] For this reason, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases containing dextran, poloxamer or a mixture thereof as an active ingredient.

[0082] Moreover, the present invention provides a method for preventing or treating inflammatory diseases including the step of administering dextran, poloxamer or a mixture thereof to a desired individual.

[0083] For this reason, specifically, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases or a method for preventing or treating inflammatory diseases containing one or more selected from the group consisting of dextran 1, dextran 5 and poloxamer 188.

[0084] In the present invention, the inflammatory disease can be an inflammatory skin disease, an inflammatory eye disease, an inflammatory osteoarticular disease, an inflammatory myopathy or an inflammatory gastrointestinal disease, but is not limited thereto.

[0085] In the case of administering two mixtures from the group consisting of the above-mentioned dextran 1, dextran 5, and poloxamer 188, a significantly excellent effect can be shown as compared with the anti-inflammatory effects of each component. Specifically, when administering dextran 1, dextran 5, and poloxamer 188, excellent anti-inflammatory effects can be obtained in all ranges of 2 (w / v)% to 40 (w / v)% of dextran 1, 2 (w / v)% to 40 (w / v)% of dextran 5, and 1 (w / v)% to 10 (w / v)% of poloxamer 188. For example, in a preferred example, by combining 5 (w / v)% of dextran 1 and 2 (w / v)% to 10 (w / v)% of poloxamer 188, 5 (w / v)% of dextran 1 and 4 (w / v)% to 40 (w / v)% of dextran 5, 50 (w / v)% of dextran 1 and 2 (w / v)% to 10 (w / v)% of poloxamer 188, 50 (w / v)% of dextran 1 and 4 (w / v)% to 40 (w / v)% of dextran 5, 2 (w / v)% of poloxamer 188 and 5 (w / v)% to 50 (w / v)% of dextran 1, 2 (w / v)% of poloxamer 188 and 4 (w / v)% to 40 (w / v)% of dextran 5, 10 (w / v)% of poloxamer 188 and 5 (w / v)% to 50 (w / v)% of dextran 1, 10 (w / v)% of poloxamer 188 and 4 (w / v)% to 40 (w / v)% of dextran 5, 4 (w / v)% of dextran 5 and 5 (w / v)% to 50 (w / v)% of dextran 1, 40 (w / v)% of dextran 5 and 5 (w / v)% to 50 (w / v)% of dextran 1, up to 40 (w / v)%, 40 (w / v)% of dextran 5 and 2 (w / v)% to 10 (w / v)% of poloxamer 188, it can be used for the therapeutic use of anti-inflammation and inflammatory diseases.

[0086] Therefore, the features of the present invention may lie in that the mixture of the present invention is a mixture of dextran 1 and poloxamer 188, the concentration of the above-mentioned dextran 1 is 2 (w / v)% to 40 (w / v)%, and the concentration of the above-mentioned poloxamer 188 is 1 (w / v)% to 10 (w / v)%.

[0087] And, the features of the present invention may lie in that the mixture of the present invention is a mixture of dextran 1 and dextran 5, the concentration of the above-mentioned dextran 1 is 2 (w / v)% to 40 (w / v)% and the concentration of the above-mentioned dextran 5 is 2 (w / v)% to 40 (w / v)%.

[0088] And, the features of the present invention may lie in that the mixture of the present invention is a mixture of dextran 5 and poloxamer 188, the concentration of the above-mentioned dextran 5 is 2 (w / v)% to 40 (w / v)%, and the concentration of the above-mentioned poloxamer 188 is 1 to 10 (w / v)%.

[0089] In another example of the present invention, it relates to a composition for regenerating cartilage based on the effect of cartilage matrix regeneration and a method for regenerating cartilage using the same.

[0090] To this end, the present invention provides a composition for regenerating cartilage containing dextran, poloxamer or a mixture thereof as an active ingredient.

[0091] In addition, the present invention provides a method for regenerating cartilage including the step of administering dextran, poloxamer or a mixture thereof to an individual in need of cartilage regeneration.

[0092] Specifically, in the present invention, there is provided a composition for regenerating cartilage or a method for regenerating cartilage containing one or more selected from the group consisting of dextran 1, dextran 5 and poloxamer 188. These induce an increase in the production amount of collagen and an increase in the production amount of proteoglycan, and can reduce the expression amount of matrix metalloproteinase-3 or matrix metalloproteinase-13.

[0093] When administering one selected from the group consisting of the above dextran 1, dextran 5 and poloxamer 188, it may contain 2 (w / v)% to 40 (w / v)% of dextran 1, 1 (w / v)% to 15 (w / v)% of poloxamer 188, and 2 (w / v)% to 30 (w / v)% of dextran 5.

[0094] In addition, when administering a mixture of two or more selected from the group consisting of dextran 1, dextran 5 and poloxamer 188, as a specific example, 5 (w / v)% of dextran 1 and 2 (w / v)% of poloxamer 188, 5 (w / v)% of dextran 1 and 4 (w / v)% to 40 (w / v)% of dextran 5, 50 (w / v)% of dextran 1 and 2 (w / v)% of poloxamer 188, 4 (w / v)% to 40% (w / v)% of dextran 5 and 2 (w / v)% to 10 (w / v)% of poloxamer 188 can be combined. In the case of combining in this way, the production amount of collagen or proteoglycan can be increased and the expression of matrix metalloproteinase-3 or matrix metalloproteinase-13 can be inhibited, thereby achieving an excellent cartilage regeneration effect.

[0095] Therefore, the present invention may be characterized in that the mixture of the present invention is a mixture of dextran 1 and poloxamer 188, the concentration of the above dextran 1 is 5 (w / v)% to 40 (w / v)%, and the concentration of the above poloxamer 188 is 2 (w / v)%.

[0096] In addition, the present invention may be characterized in that the mixture of the present invention is a mixture of dextran 1 and dextran 5, the concentration of the above dextran 1 is 5 (w / v)%, and the concentration of the above dextran 5 is 4 (w / v)% to 40 (w / v)%.

[0097] Moreover, the characteristics of the present invention may lie in that the mixture of the present invention is a mixture of dextran 5 and poloxamer 188, the concentration of the above dextran 5 is 4 (w / v)% to 40 (w / v)%, and the concentration of the above poloxamer 188 is 2 (w / v)% to 10 (w / v)%.

[0098] Moreover, the anti-inflammatory and cartilage regeneration effects can be achieved simultaneously. Therefore, examples most suitable for the prevention or treatment of joint diseases or connective tissue diseases include, but are not limited to, in the case of using dextran 1, dextran 5, and poloxamer 188 alone, dextran 1 can be 2.5 (w / v)%, dextran 5 can be 2 (w / v)% to 30 (w / v)%, and poloxamer 188 can be 1 (w / v)% to 15 (w / v)%. In the case of using a mixture of two or more selected from the group consisting of dextran 1, dextran 5, and poloxamer 188, 5 (w / v)% of dextran 1 and 4 (w / v)% to 40 (w / v)% of dextran 5, 4 (w / v)% to 40 (w / v)% of dextran 5 and preferably 2 (w / v)% to 10 (w / v)%, 2 (w / v)% of poloxamer 188 and 5 (w / v)% to 40 (w / v)% of dextran 1 are used in combination.

[0099] Terms not otherwise defined in the present invention have the meanings commonly used in the technical field to which the present invention pertains. Detailed Description of the Invention

[0101] Materials and Methods

[0102] In this application, experiments were conducted using dextran 1 (Dextran 1 (EP grade, Pharmacosmos)), dextran 5 (Dextran5, pharmaceutical quality, Pharmacosmos), and poloxamer 188 (Poloxamer 188, cell culture grade, Sigma-Aldrich). Moreover, to induce arthritis, recombinant human interleukin-1α (recombinant human IL-1α, Lot No. 200-01A) was purchased from PeproTech, and indomethacin (Indomethacin, Sigma-Aldrich, Lot No. 53-86-1) and sodium hyaluronate (25 mg / 2.5 mL of sodium hyaluronate, Aragan Injection, Dongkwang Pharm, proprietary medicine) were used as positive control substances.

[0103] Example 1. Preparation and Experimental Preparation of Dextran, Poloxamer or Their Mixtures

[0104] 1.1 Isolation and Culture of Cells

[0105] After isolating the hind leg joints of 3-week-old male Sprague-Dawley rats under sterile conditions, only the cartilage tissue constituting the joints was collected, and it was made into single cells to isolate primary rat chondrocytes for use. When the cells in primary culture were stable, they were used in the experiment. To determine stable chondrocytes, after isolating ribonucleic acid (RNA), the expression levels of type II collagen or SOX9 gene, which are chondrocyte marker factors, were confirmed by real-time reverse transcription-polymerase chain reaction (RT-PCR) and then used in the experiment.

[0106] 1.2 Cell Culture Method

[0107] The cells isolated in 1.1 were cultured in an incubator set at a temperature of 37°C, a humidity of 95%, and 5% CO 2 . The temperature and humidity of the culture chamber were confirmed every 8 hours. The culture medium used was Minimum Essential Medium (MEM) supplemented with 10% fetal bovine serum, 2 mL of L-glutamine, 50 U / mL of penicillin, and 50 μg / mL of streptomycin. During the culture period, when the cell number proliferated to more than 90%, the cells were separated using a detached solution (0.25 (w / v)% trypsin, 0.53 mM ethylenediaminetetraacetic acid solution (EDTA solution) (3 mL)), and the cells were separated by centrifugation at 125 Xg for 10 minutes for use in the following experiments.

[0108] 1.3 Composition and Preparation of Test Groups

[0109] The test groups were formed by treating a model induced with osteoarthritis by 5 ng / mL of recombinant human interleukin-1α with a normal control group (NC), an inflammation-induced control group (PC), and a test substance (single substance or mixture) at different concentrations. The indomethacin (IM) or sodium hyaluronate (HN) treatment groups as positive control groups were formed with a single concentration. The test groups are shown in Table 1 below.

[0110] Table 1

[0111]

[0112]

[0113]

[0114] D = dextran 1, T = dextran 5, P = poloxamer 188, NC = normal group, PC = group administered with an inflammation inducer (recombinant human interleukin-1α), HN = sodium hyaluronate, IM = indomethacin, D+P = mixture of dextran 1 and poloxamer 188, D+T = mixture of dextran 1 and dextran 5, P+T = mixture of poloxamer 188 and dextran 5.

[0115] The concentrations of the components used in the present invention are labeled according to the following labeling method: D10 = 10 (w / v)% solution of dextran 1, P10 = 10 (w / v)% solution of poloxamer 188, T10 = 10 (w / v)% solution of dextran 5.

[0116] The test substances used in the present invention are prepared by the following method: The maximum concentration dissolved in cell culture medium is reflected, and single substances are prepared in the range of 1 to 50 (w / v)% for dextran 1 (D), 1 to 40 (w / v)% for dextran 5 (T), and 1 to 35 (w / v)% for poloxamer 188 (P). In the case of preparing a mixture, two single substances are mixed at a volume ratio of 1:1 (v:v).

[0117] In a complete medium (complete media) (minimum essential medium containing all 10% fetal bovine serum, 2 mL of L-glutamine, 50 U / mL of penicillin, and 50 μg / mL of streptomycin) prepared for cell culture, dextran 1, dextran 5, and poloxamer 188 are completely dissolved according to the administration concentration (w / v). The test substance is completely dissolved in the complete medium for homogenization. At low temperature, especially in the case of poloxamer 188, at a lower temperature, it is slowly stirred with a sterile spatula, stick, magnetic bar, etc., or gently shaken several times to the extent that no bubbles are formed until it is completely transparent without suspended matter. After the test substance is completely dissolved, it is filtered through a 0.22 μm pore size syringe filter, sealed, and stored refrigerated.

[0118] More specifically, the preparation of the single substance is carried out by the following method.

[0119] Prepare a 5% single substance of dextran 1 ('D5')

[0120] In the case of preparing a 5% test solution of dextran 1, 5 g of dextran 1 was taken respectively and completely dissolved in a complete medium and made up to 100 mL to prepare a 5 (w / v)% D test solution. The test substance dissolved at a concentration of 0.05 g / mL was filtered through a syringe filter with a pore size of 0.22 μm, refrigerated and stored for later use.

[0121] Preparation of a mixture of 5% dextran 1 and 10% poloxamer 188 ('D5+P10')

[0122] D5 and P10 were prepared separately, filtered through a syringe filter with a pore size of 0.22 μm, refrigerated and stored. Then, each volume was mixed at a ratio of 1:1 (v:v), shaken gently at a low speed to avoid foaming, and stored for later use.

[0123] Comparative example: Preparation of indomethacin-treated group (IM)

[0124] Indomethacin was completely dissolved in dimethyl sulfoxide (DMSO) at a concentration of 5 mM, diluted with a complete medium, filtered through a syringe filter with a pore size of 0.22 μm, and the final concentration was adjusted to 5 μM when treating cells.

[0125] Comparative example: Preparation of sodium hyaluronate-treated group (HN)

[0126] Sodium hyaluronate was aspirated into a complete medium in an Aragorn syringe (25 mg / 2.5 mL of sodium hyaluronate, in a pharmaceutical form packed in a disposable sterile syringe), diluted with 25 mg / 3 mL of sodium hyaluronate (= 8.33 mg / mL of sodium hyaluronate), and used directly.

[0127] 1.4 Set the administration method

[0128] In the case of primary cultured chondrocytes isolated from rats, the doubling time was approximately 24 hours. Therefore, the test substance was treated when the proliferation and stability were appropriate. After culturing chondrocytes in a 48-well or 24-well plate, each test substance was administered once. One hour before treatment with recombinant human interleukin-1α for inducing arthritis, the test solution was added according to the concentration of each test substance so that the concentration was 20% (v / v) of the total culture medium volume. After treatment with recombinant human interleukin-1α for 24 hours, the culture medium was taken for analysis experiments. However, when confirming the toxicity or proliferation ability of the test substance on chondrocytes, recombinant human interleukin-1α was not treated, and only the test substance was treated.

[0129] 1.5 Statistical analysis

[0130] All experimental results were analyzed by t - test (student T - test) and compared with the normal control group (NC) or the positive control group for inflammation induction (PC) at the p < 0.05 level, and significance was shown.

[0131] Example 2. Cytotoxicity analysis

[0132] Cytotoxicity analysis was performed by confirming whether the proliferative ability of chondrocytes changed according to the treatment with the test substance. The evaluation of cell proliferative ability was performed in both untreated chondrocytes without recombinant human interleukin - 1α treatment and chondrocytes induced to inflammation by treatment with recombinant human interleukin - 1α. After treating each test substance on rat chondrocytes and culturing for 24 hours and 48 hours, thiazolyl blue tetrazolium blue assay (MTT; Sigma, M5655) was then carried out. In the evaluation of the cell proliferative ability of the test substance in an in vitro model of arthritis induced by recombinant human interleukin - 1α as an inflammation - inducing substance, 1 hour before the treatment with recombinant human interleukin - 1α, each test substance was treated on rat chondrocytes and cultured for 24 hours and 48 hours, and then thiazolyl blue tetrazolium blue assay (Sigma, M5655) was carried out. The results are as Figures 1 to 4 shown.

[0133] As Figure 1 shown, in the knee joint chondrocytes of rats without induced inflammation, the results of observing the cell proliferation rate of dextran 1, dextran 5, poloxamer 188 or their mixture for 24 hours confirmed that most of dextran 1, dextran 5, poloxamer 188 and their mixture were not toxic to cells and contributed to cell proliferation. However, in the case of the poloxamer 188 treatment group in the single - treatment group, when poloxamer 188 was cultured at concentrations of 25 (w / v)% (P25) and 35 (w / v)% (P35) for 24 hours, the reduction of partial cell proliferative ability was lower than that of the sodium hyaluronate (HN) administration group as a control drug (commercially available drug), so it was not suitable for single administration. And in their mixture, when cultured for 24 hours, a reduction of partial cell proliferative ability occurred, from which it was confirmed that in the case of poloxamer 188, when treated at a high concentration, it could induce cytotoxicity. And in the mixture, in the D50 + P35, P5 + T40, P25 + T40, P35 + T40 experimental groups, the reduction of cell proliferative ability was lower than that of the sodium hyaluronate (HN) administration group as a control drug (commercially available drug).

[0134] As Figure 2 shown, the results of observing the cell proliferation rate of dextran, poloxamer or their mixture for 48 hours in the knee joint chondrocytes of rats without induced inflammation also confirmed that the results were similar to those of culturing for 24 hours.

[0135] These results confirmed that when treating normal knee joint chondrocytes, even when the concentration of dextran 1 (D) was increased to 50% or the concentration of dextran 5 (T) was increased to 40%, the proliferation of chondrocytes was not inhibited compared to commercially available drugs. On the contrary, in the case of poloxamer 188, at concentrations above 25%, it can be toxic to chondrocytes. And, in the case of being formed by a mixture, compared with poloxamer 188 alone, the cell proliferation ability was partially improved. However, in order not to induce toxicity, preferably, poloxamer 188 was mixed at a concentration of 10 (w / v)% or less.

[0136] In Figure 3 and Figure 4 showed the results of observing the cell proliferation rates of the experimental groups according to the treatment for 24 hours and 48 hours respectively in an in vitro model of osteoarthritis treated with recombinant human interleukin-1α.

[0137] As Figure 3 shown, in the osteoarthritis model, poloxamer 188 also inhibited cell proliferation when the concentration increased, while dextran 1 and dextran 5 did not inhibit cell proliferation even when the concentration increased. In the case of poloxamer 188, cell proliferation was confirmed to be inhibited in the P10 single treatment group. However, when it was used in combination with D5, D50, T4, T40, etc., it had a cell proliferation effect. When treated in combination with dextran, 1 (w / v)% to 10 (w / v)% of poloxamer 188 could be used.

[0138] In Figure 4 the same results as those for culturing for 24 hours could be confirmed, and it was confirmed that the cell proliferation ability of the P10 treatment group was increased by mixing with dextran 1 or dextran 5.

[0139] Based on the above results, it was confirmed that dextran 1 and dextran 5 did not induce cytotoxicity in normal and inflammation-induced cells. On the contrary, with the increase in the concentration of poloxamer 188, when treated with a single substance, it could induce inhibition of cell proliferation ability in both normal cells and inflammation-induced cells. When 1 (w / v)% to 10 (w / v)% of poloxamer 188 was administered in combination with dextran 1 or dextran 5, it could be used without cytotoxicity.

[0140] Example 3. Confirming the anti-inflammatory effect by the interleukin-10 / interleukin-6 expression ratio

[0141] Experiments were conducted to confirm the anti-inflammatory effects of dextran 1, dextran 5, or poloxamer 188. Specifically, single administration or combined administration was carried out by changing dextran 1 to 2.5 (w / v)% to 50 (w / v)%, dextran 5 to 2 (w / v)% to 40 (w / v)%, and poloxamer 188 to 1 (w / v)% to 20 (w / v)%, and the interleukin-10 / interleukin-6 expression ratio based thereon was confirmed. To induce inflammation, recombinant human interleukin-1α, an inflammatory inducer, was treated, and the interleukin-10 / interleukin-6 expression ratio when recombinant human interleukin-1α was treated together with the test substances (single substances, their mixtures) was confirmed. It can be judged that the higher the interleukin-10 / interleukin-6 expression ratio, the better the anti-inflammatory effect.

[0142] The composition of the primers used in the experiment is shown in Table 2 below. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a housekeeping gene, was used as a control group. The results are shown in Tables 3a to 3b and Figure 5 as shown

[0143] Table 2

[0144]

[0145]

[0146] After mixing the primer, SyBr green (SYBR Premix Ex Taq, Takara, RR420A), and the template, real-time reverse transcription-polymerase chain reaction (Real time RT-PCR) (CFX 96 touch, Bio rad, Hercules, CA, USA) was performed based on the intercalating method. The quantitative results (gene expression levels) obtained by applying the Ct value to the calibration curve were expressed as the value divided by the quantitative result value of glyceraldehyde-3-phosphate dehydrogenase, a housekeeping gene.

[0147] Table 3a

[0148]

[0149]

[0150] The interleukin-10 / interleukin-6 gene expression ratio of single substances of dextran 1 (D), dextran 5 (T), and poloxamer 188 (P) in chondrocytes treated with recombinant human interleukin-1α for 24 hours

[0151] Table 3b

[0152]

[0153]

[0154] Interleukin-10 / interleukin-6 gene expression ratio of dextran 1 (D), dextran 5 (T), or poloxamer 188 (P) mixture in chondrocytes treated with recombinant human interleukin-1α for 24 hours

[0155] As shown in Table 3a and Figure 5 As shown, in the group treated with dextran 1 (D) alone as a single-group treatment experiment, an increase in the interleukin-10 / interleukin-6 expression ratio was not clearly confirmed except for the 50 (w / v)% treatment group. However, in the 1 (w / v)% to 20 (w / v)% poloxamer 188 (P) treatment group and the 2 (w / v)% to 20 (w / v)% dextran 5 (T) group, a significant increase effect was confirmed.

[0156] As shown in Table 3b and Figure 5 As shown, in the mixture administration group of dextran 1 (D) in the mixture treatment experiment, a significant increase effect in the interleukin-10 / interleukin-6 expression ratio was observed, and a significant increase was also observed in the combined administration group of poloxamer 188 (P) and dextran 5 (T). Therefore, the anti-inflammatory efficacy in the mixture was confirmed to be excellent.

[0157] In the 50 (w / v)% dextran 1 administration group, the increase in the interleukin-10 / interleukin-6 expression ratio was significant. However, considering that the increase amount was not significant, the increase in the interleukin-10 / interleukin-6 expression ratio in the osteoarthritis model, that is, the anti-inflammatory effect was better when administered by mixing with poloxamer 188 (P) or dextran 5 (T) compared to increasing the concentration of dextran 1 (D). In particular, compared with the single administration groups of 5 (w / v)% to 50 (w / v)% dextran 1 (D), 4 (w / v)% to 40 (w / v)% dextran 5 (T), and 2 (w / v)% to 10 (w / v)% poloxamer 188 (P), the anti-inflammatory effects of their mixture administration groups were mostly significantly excellent. Thus, the excellence of the combined administration of dextran 1 (D), dextran 5 (T), and poloxamer 188 (P) was confirmed.

[0158] In particular, the combined administration of dextran 1 (D), dextran 5 (T), and poloxamer 188 (P) has excellent anti-inflammatory effects within all ranges of 5 (w / v)% to 50 (w / v)% dextran 1 (D), 4 (w / v)% to 40 (w / v)% dextran 5 (T), and 2 (w / v)% to 10 (w / v)% poloxamer 188 (P). All of these combinations have anti-inflammatory effects, and thus, it is confirmed that they are effective in the treatment of inflammatory diseases.

[0159] Example 4. Confirmation of cartilage matrix regeneration effect by changes in matrix metalloproteinase-3 and matrix metalloproteinase-13 gene expression

[0160] An experiment for confirming the therapeutic effect of osteoarthritis was conducted by gene expression of an index substance for evaluating osteoarthritis. Matrix metalloproteinase-3 and matrix metalloproteinase-13 are proteins that destroy the cartilage matrix. Therefore, if the increase in the expression level of the above genes, which can be induced by inflammatory stimuli, can be effectively inhibited, an excellent cartilage protection effect can be evaluated.

[0161] The expression changes of matrix metalloproteinase-3 and matrix metalloproteinase-13 based thereon were confirmed by single administration or combined administration by changing dextran 1 to 2.5 (w / v)% to 50 (w / v)%, dextran 5 to 2 (w / v)% to 40 (w / v)%, and poloxamer 188 to 1 (w / v)% to 20 (w / v)%. Specifically, 1 hour before the treatment with recombinant human interleukin-1α, each test substance was treated on rat chondrocytes and then cultured for 24 hours. The supernatant of the cultured cells was removed, and the cells were washed with phosphate buffer solution (PBS). Ribonucleic acid was isolated from the washed cells (GeneAll hybrid-R RNA purification kit; GeneAll, 3033522), and the isolated ribonucleic acid was quantified using a nanodrop (Take3 Multi-Volume plate, BioTeK, Instruments, VT, USA) and then diluted to the same concentration for each experimental group. The ribonucleic acid diluted to an equal amount was synthesized into complementary deoxyribonucleic acid (cDNA) by polymerase chain reaction (ReverTraAceR qPCR RT Master Mix with gDNA Remover, Toyobo, FSQ-301). Using the synthesized complementary deoxyribonucleic acid, the gene expression of matrix metalloproteinase-3 and matrix metalloproteinase-13 was confirmed in a 96-well plate by real-time reverse transcription-polymerase chain reaction.

[0162] The structures of the primers used in the experiment are shown in Table 4 below, and glyceraldehyde-3-phosphate dehydrogenase, which is a housekeeping gene, was used as a control group.

[0163] Table 4

[0164]

[0165]

[0166] After mixing the primer, SyBr green (SYBR Premix Ex Taq, Takara, RR420A), and the template, real-time reverse transcription-polymerase chain reaction based on the intercalation method was performed using a real-time reverse transcription-polymerase chain reaction (CFX 96 touch, Bio rad, Hercules, CA, USA). By applying the Ct value The quantitative result (gene expression level) obtained by applying it to the calibration curve is expressed as the value obtained by dividing this value by the quantitative result value of glyceraldehyde-3-phosphate dehydrogenase as a housekeeping gene.

[0167] 3.1 Confirmation of changes in matrix metalloproteinase-3 expression

[0168] The expression results of matrix metalloproteinase-3 treated with dextran 1, dextran 5, poloxamer 188 alone or their mixture are shown in Tables 5a to 5b and Figure 6 in.

[0169] Table 5a

[0170]

[0171]

[0172] Chondrocytes treated with recombinant human interleukin-1α for 24 hours

[0173] The matrix metalloproteinase-3 gene expression levels of dextran 1 (D), dextran 5 (T), and poloxamer 188 (P) alone in

[0174] Table 5b

[0175]

[0176]

[0177] The matrix metalloproteinase-3 gene expression levels of the mixture of dextran 1 (D), dextran 5 (T), or poloxamer 188 (P) in chondrocytes treated with recombinant human interleukin-1α for 24 hours

[0178] As shown in Table 5a above and Figure 6As shown, compared with the positive control group induced by inflammation, dextran 1 (D) has a significant effect of reducing matrix metalloproteinase-3 at concentrations from 2.5 (w / v)% to 40 (w / v)%, while dextran 5 (T) has a significant effect of reducing matrix metalloproteinase-3 at concentrations from 2 (w / v)% to 20 (w / v)%. On the other hand, poloxamer 188 (P) has a reducing effect at 1 (w / v)% to 20 (w / v)% in all groups where the experiment was conducted.

[0179] As shown in Table 5b and Figure 6 As shown, in the experimental group of the mixture, it was confirmed that there was a significant effect in all experimental groups of 5 (w / v)% dextran 1 (D) and 2 (w / v)% to 10 (w / v)% poloxamer 188 (P), 4% to 40 (w / v)% dextran (T). However, in the group treated with dextran 1 (D) at a high concentration of up to 50 (w / v)%, a significant reduction effect was only confirmed when mixed with P2. Therefore, when mixing dextran 1 with dextran 5 or poloxamer 188, preferably, the mixing is carried out at a concentration of less than 50 (w / v)%. Poloxamer 188 has a significant effect of reducing matrix metalloproteinase-3 in most of the single treatment and mixed treatment groups. However, it was confirmed in Example 2 that preferably, the concentration of poloxamer is 10 (w / v)% or less, and significant matrix metalloproteinase-3 inhibitory effects can also be induced in the combinations of corresponding P2, P5, P10 with dextran 1 and dextran 5.

[0180] 3.2 Confirmation of changes in matrix metalloproteinase-13 expression

[0181] The expression results of matrix metalloproteinase-13 treated with dextran 1, dextran 5, poloxamer 188 alone or their mixtures are shown in Tables 6a to 6b and Figure 7 in.

[0182] Table 6a

[0183]

[0184] The gene expression levels of matrix metalloproteinase-13 of dextran 1 (D), dextran 5 (T), and poloxamer 188 (P) alone in chondrocytes treated with recombinant human interleukin-1α for 24 hours

[0185] Table 6b

[0186]

[0187]

[0188] Matrix metalloproteinase-13 gene expression levels of the mixture of dextran 1 (D), dextran 5 (T), or poloxamer 188 (P) in chondrocytes treated with recombinant human interleukin-1α for 24 hours

[0189] From Tables 6a to 6b above, Figure 7 it can be confirmed that in the single-substance experimental group, dextran 1 (D) has a significant effect of inhibiting the increase in matrix metalloproteinase-13 gene at concentrations from 2.5 (w / v)% to 25 (w / v)%. D40 has no significant effect at a concentration of 40 (w / v)%, but the increase in matrix metalloproteinase-13 gene is inhibited. Instead, at a concentration of 50 (w / v)%, the matrix metalloproteinase-13 level increases for D50. A similar property also exists in the mixture of dextran 1 (D). Therefore, it is confirmed that the most preferred concentration range of dextran 1 (D) is from 2.5 (w / v)% to 40 (w / v)%, which is consistent with the results of matrix metalloproteinase-3 gene expression levels. When dextran 5 (T) is administered alone, it has a significant effect of inhibiting the increase in matrix metalloproteinase-13 gene at concentrations from 2 (w / v)% to 40 (w / v)%. Dextran 5 (T) at 4 (w / v)% and 40 (w / v)% also has a significant effect of inhibiting the increase in matrix metalloproteinase-13 gene in the mixture with 5 (w / v)% of dextran 1 (D) and 2 (w / v)% to 10 (w / v)% of poloxamer 188 (P).

[0190] In the case of poloxamer 188, it has an effect of inhibiting the increase in matrix metalloproteinase-13 gene at concentrations from 5 (w / v)% to 20 (w / v)% in the single experimental group. However, when administered as a mixture with dextran, it is confirmed that it can also have an effect at a concentration of 2 (w / v)%. Considering the cytotoxicity experimental results in Example 2, preferably, the concentration of poloxamer 188 (P) is from 2 (w / v)% to 10 (w / v)% when administered in combination. Except for the combination with 50 (w / v)% of dextran 1 (D), the corresponding combinations of P2, P10 with dextran 1 and dextran 5 also have the effect of inducing the inhibition of the increase in matrix metalloproteinase-13 gene.

[0191] Based on the above results, significant matrix metalloproteinase-13 inhibitory effects are also induced in the combinations of P2, P5, P10 with dextran 1 and dextran 5.

[0192] Example 5. Confirming the increase in the production of type II collagen and aggrecan in the osteoarthritis model

[0193] To confirm the production amounts of type II collagen and aggrecan in the indicator substances for evaluating osteoarthritis, each test substance was treated on chondrocytes and cultured for 24 hours before the treatment with recombinant human interleukin-1α for 1 hour. Since type II collagen and aggrecan are components of the cartilage matrix, if their production amounts increase, it can be determined that the cartilage matrix protection effect is excellent.

[0194] Among them, the cell supernatant (culture medium) was recovered and centrifuged at 13,000 rpm for 10 minutes, and only the supernatant was used for the analysis. Using the centrifuged cell supernatant, the analysis was carried out according to the manuals of the Aggrecan ELISA Kit (Rat Aggrecan ELISA Kit, Mybiosource, MBS261073) and the type II collagen (Type II Collagen detection Kit, Multi-Species, Chondrex, 6018) products. To perform accurate analysis, an ELISA reader (EPOCH 2 microplate reader, BioTek, Instruments, VT, USA) with the full wavelength was used to measure the absorbance at 450 nm and 490 nm respectively, and then the contents (ng / mL) of type II collagen and aggrecan were quantified according to the calibration curve.

[0195] 5.1 Confirm the production amount of type II collagen

[0196] The results of the production amounts of type II collagen according to dextran 1 (D), dextran 5 (T), poloxamer 188 (P) alone or their mixtures are shown in Tables 7a to 7b and Figure 8 in.

[0197] Table 7a

[0198]

[0199]

[0200] The production amounts of type II collagen of dextran 1 (D), dextran 5 (T), and poloxamer 188 (P) alone in chondrocytes treated with recombinant human interleukin-1α for 24 hours

[0201] Table 7b

[0202]

[0203]

[0204] Amount of type II collagen produced by chondrocytes treated with recombinant human interleukin-1α for 24 hours in a mixture of dextran 1 (D), dextran 5 (T), or poloxamer 188 (P)

[0205] As shown in Tables 7a to 7b above and Figure 8 As shown, an increase in type II collagen synthesis was observed in the groups administered dextran 1 (D) alone at 5 (w / v)% to 25 (w / v)% and dextran 5 (T) alone at 4 (w / v)% to 40 (w / v)% relative to the positive control group for inflammation induction (PC). On the other hand, relative to the positive control group for inflammation induction (PC), neither the administration of dextran 1 (D) alone at 50 (w / v)% nor the combined administration with dextran 5 (T) and poloxamer 188 (P) induced an increase in type II collagen synthesis. This indicates that, as confirmed in Example 4, 50 (w / v)% dextran 1 (D) alone does not have the effect of suppressing the increased expression of matrix metalloproteinase-3 and matrix metalloproteinase-13. Considering that only the combination with 2 (w / v)% to 10 (w / v)% poloxamer 188 (P) can suppress the increased expression of matrix metalloproteinase-3 and matrix metalloproteinase-13, preferably, dextran 1 (D) is used alone or in combination at a concentration of 40 (w / v)% or less. In particular, when 5 (w / v)% dextran 1 (D) and 4 (w / v)% to 40 (w / v)% dextran 5 (T) are administered in combination, a significant increase in type II collagen synthesis is observed relative to the positive control group for inflammation induction (PC).

[0206] In the case of poloxamer 188 (P), no increase in type II collagen synthesis was observed in the single-treatment groups at 10 (w / v)% and 20 (w / v)% except for the 2 (w / v)% treatment group, and it was confirmed that type II collagen synthesis further decreased with increasing concentration. However, when 2 (w / v)% or 10 (w / v)% poloxamer 188 (P) was administered in combination with 5 (w / v)% dextran 1 (D) or 40 (w / v)% dextran 5 (T), a significant increase in type II collagen synthesis was observed. These results indicate that in the synthesis of type II collagen in osteoarthritis, dextran 1 (D) and dextran 5 (T) can be treated alone, but except for 2 (w / v)% poloxamer 188 (P), poloxamer 188 (P) is not suitable for single treatment, and the combined administration of poloxamer 188 (P) with dextran 1 (D) or dextran 5 (T) can effectively induce type II collagen synthesis.

[0207] In Example 3, it was confirmed that compared with single administration, the combined administration of poloxamer 188 and dextran 1 or dextran 5 can significantly increase the gene expression ratio of interleukin-10 / interleukin-6. In summary, in order to alleviate the inflammatory response in osteoarthritis and promote collagen synthesis, preferably, a mixture of dextran 1, dextran 5 or poloxamer 188 is administered.

[0208] 5.2 Confirm the quantitative results of proteoglycan content

[0209] The results of proteoglycan production treated with dextran 1, dextran 5, poloxamer 188 alone or their mixtures are shown in Tables 8a to 8b and Figure 9 in.

[0210] Table 8a

[0211]

[0212] Table 8b

[0213]

[0214]

[0215] Proteoglycan production of the mixture of dextran 1 (D), dextran 5 (T), and poloxamer 188 (P) in chondrocytes treated with recombinant human interleukin-1α for 24 hours

[0216] It can be confirmed from the above Tables 6a and Figure 9 that, compared with the positive control group for inflammation induction (PC), in the groups treated with 5 (w / v)% to 25 (w / v)% dextran 1 (D), 4 (w / v)% dextran 5 (T), and 2 (w / v)% to 10 (w / v)% poloxamer 188 (P) alone, the proteoglycan production increased significantly. On the other hand, the groups administered 50 (w / v)% dextran 1 (D), 20 (w / v)% poloxamer 188 (P), and 20 (w / v)% to 40 (w / v)% dextran 5 (T) alone did not induce an increase in proteoglycan production.

[0217] From Table 8b and Figure 9It can be confirmed that, compared with the inflammation-induced positive control group (PC), in the mixed administration groups, the production amount of proteoglycan was significantly increased in the mixed administration groups of 5 (w / v)% dextran 1 (D) and 2 (w / v)% poloxamer 188 (P), or 4 (w / v)% to 40 (w / v)% dextran 5 (T). The production amount of proteoglycan was significantly increased only in the mixed administration group of 50 (w / v)% dextran 1 (D) and 2 (w / v)% poloxamer 188 (P). Compared with the inflammation-induced positive control group (PC), the production amount of proteoglycan was also significantly increased in the mixed administration groups between 4 (w / v)% to 40 (w / v)% dextran 5 (T) and 2 (w / v)% to 10 (w / v)% poloxamer 188 (P). In particular, compared with the single administration of 5 (w / v)% dextran 1 (D) and 2 (w / v)% poloxamer 188 (P), the production amount of proteoglycan was significantly increased when mixed with 4 (w / v)% to 40 (w / v)% dextran 5 (T).

[0218] Considering together the results of type II collagen synthesis confirmed in Example 5.1, in the osteoarthritis model, in order to increase type II collagen synthesis and increase the production of proteoglycan, preferably, 5 (w / v)% to 25 (w / v)% dextran 1 (D), 4 (w / v)% dextran 5 (T), or 2 (w / v)% poloxamer 188 (P) is administered alone, or 5 (w / v)% dextran 1 (D) is mixed with 4 (w / v)% to 40 (w / v)% dextran 5 (T), or 5 (w / v)% dextran 1 (D) is mixed with 2 (w / v)% poloxamer 188 (P). And in the combination of 4 (w / v)% to 40 (w / v)% dextran 5 (T) and 5 (w / v)% dextran 1 (D), in particular, in the case of 40 (w / v)% dextran 5, in the combination with 2 (w / v)% to 10 (w / v)% poloxamer 188 (P), the synthesis of type II collagen and the production of proteoglycan can be significantly increased.

[0219] In summary, in the treatment of osteoarthritis, dextran 1, dextran 5 and poloxamer 188 can be used, but not all of these alone or in all combinations are effective for the treatment of osteoarthritis. Depending on the concentration combination of dextran 1, dextran 5 and poloxamer 188, the toxicity and effect may be different. Therefore, it can be confirmed that it is important to obtain an appropriate combination thereof. Sequence Listing <110> MEDICINE PARK CO., LTD <120> Composition for treating joint diseases or connective tissue diseases containing dextran or poloxamer <130> LIH1.1 <150> KR 10-2017-0165271 <151> 2017-12-04 <160> 10 <170> PatentIn version 3.2 <210> 1 <211> 19 <212> DNA <213> Artificial sequence <220> <223> MMP-3 forward primer <400> 1 tgggaagcca gtggaaatg 19 <210> 2 <211> 21 <212> DNA <213> Artificial sequence <220> <223> MMP-3 reverse primer <400> 2 ccatgcaatg ggtaggatga g 21 <210> 3 <211> 20 <212> DNA <213> Artificial sequence <220> <223> MMP-13 forward primer <400> 3 ctgacctggg atttccaaaa 20 <210> 4 <211> 20 <212> DNA <213> Artificial sequence <220> <223> MMP-13 reverse primer <400> 4 acacgtggtt ccctgagaag 20 <210> 5 <211> 26 <212> DNA <213> Artificial sequence <220> <223> GAPDH forward primer <400> 5 ctcaactaca tggtctacat gttcca 26 <210> 6 <211> 22 <212> DNA <213> Artificial sequence <220> <223> GAPDH reverse primer <400> 6 cttcccattc tcagccttga ct 22 <210> 7 <211> 20 <212> DNA <213> Artificial sequence <220> <223> IL-6 forward primer <400> 7 tgatggatgc ttccaaactg 20 <210> 8 <211> 20 <212> DNA <213> Artificial sequence <220> <223> IL-6 reverse primer <400> 8 gagcattgga agttggggta 20 <210> 9 <211> 21 <212> DNA <213> Artificial sequence <220> <223> IL-10 forward primer <400> 9 gttgccaagc cttgtcagaa a 21 <210> 10 <211> 20 <212> DNA <213> Artificial sequence <220> <223> IL-10 reverse primer <400> 10 tttctgggcc atggttctct 20

Claims

1. Use of a composition containing dextran 5 as an active ingredient in the preparation of a medicament for treating osteoarthritis; wherein the composition contains dextran 5 as the sole dextran, and the concentration of dextran 5 is 2 w / v% to 30 w / v%.

2. Use of a composition containing dextran 5 and dextran 1 as active ingredients in the preparation of a medicament for treating osteoarthritis, wherein, i) the concentration of dextran 5 is 2 w / v% to 40 w / v%, and ii) the concentration of dextran 1 is 2.5 w / v% to 40 w / v%.

3. The use according to claim 2, wherein the w / v% concentration ratio of dextran 1 and dextran 5 in the composition is 1:0.01 to 1:

20.

4. Use of a composition containing dextran 5 and poloxamer as active ingredients in the preparation of a pharmaceutical composition for treating osteoarthritis, wherein the concentration of dextran 5 is 2 w / v% to 40 w / v%; the concentration of the poloxamer is 1 w / v% to 10 w / v%.

5. The use according to claim 4, wherein the w / v% concentration ratio of dextran 5:poloxamer in the composition is 1:0.01 to 1:

40.

6. The use according to claim 4, wherein the poloxamer has an average molecular weight of 100 Da to 20000 Da.

7. The use according to claim 4, wherein the poloxamer is selected from the group consisting of poloxamer 101, poloxamer 105, poloxamer 105 benzoate, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 182 dibenzoate, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, poloxamer 288, poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403 and poloxamer 407.

8. The use according to claim 1, 2 or 4, wherein the composition further comprises stem cells.

9. Use of a composition containing dextran 5 as an active ingredient in the preparation of a medicament for cartilage regeneration; wherein the composition contains dextran 5 as the sole dextran, and the concentration of dextran 5 is 2 w / v% to 30 w / v%.

10. Use of a composition containing dextran 5 and dextran 1 as active ingredients in the preparation of a medicament for cartilage regeneration, wherein, the concentration range of dextran 5 is 2 w / v% to 40 w / v%, and the concentration range of dextran 1 is 2.5 w / v% to 40 w / v%.

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