KR20210093166A - Composition for regeneration of tissue - Google Patents

Abstract

The present invention induces and promotes differentiation of stem cells into cartilages and inhibits degradation of proteoglycans and glycosaminoglycans in cartilage cells in an inflammatory environment by using a cartilage-binding peptide or a nucleic acid sequence encoding the same and an adeno-associated virus (AAV) vector, together with an activity inhibitor of casein kinase 2 (CK2) protein or an expression inhibitor of a gene encoding the CK2 protein. Accordingly, cartilage regeneration is possible and also cartilage-related diseases can be effectively prevented, alleviated, or treated.

Description

Composition for regeneration of tissue

The present invention relates to a composition for tissue, particularly cartilage regeneration, and a composition for preventing, improving or treating cartilage disease using the same.

Cartilage, a tissue with elasticity, connects the two bones to each other, gives mobility to the joint area, and serves to cushion the impact. In particular, articular cartilage is a connective tissue, and such cartilage includes chondrocytes embedded in a matrix of proteoglycan and type collagen. When cartilage tissue is damaged by various causes, direct friction between bone and bone, stiffness of the joint, gradual decrease in articular cartilage movement, and frequent pain in the cartilage area are induced. According to the findings so far, it is known that the initial pathogenesis of osteoarthritis is usually caused by cartilage destruction due to changes or imbalances in anabolic and catabolic metabolic mechanisms in the articular cartilage matrix.

In general, the cartilage tissue constituting the joints of vertebrates lacks blood vessels, nerves, and lymphoid tissue, and once the cartilage tissue is damaged, it cannot be normally regenerated in vivo. If the cartilage tissue of the joint is damaged in this way, daily activities are restricted due to severe pain, and if the cartilage tissue damage becomes chronic, it causes fatal degenerative arthritis, making it impossible to perform normal life or professional activities. Accordingly, various medical methods for regenerating damaged cartilage tissue have been tried, but it is still insufficient to restore the articular cartilage, Hyaline cartilage, to a natural state.

Clinically, ① a method of inducing differentiation of stem cells into chondrocytes, ② a method of transplanting autologous or allogeneic cartilage tissue into a cartilage defect site by osteochondral transplantation, ③ a method of inducing the differentiation of stem cells into chondrocytes, ③ microfracture When the cartilage is scraped off and removed, and the subchondral bone is exposed, a hole is drilled at regular intervals to promote the regeneration of bone marrow cells that have flowed through it into cartilage tissue, and ④ Chondrocyte transplantation is used to repair the cartilage defect site. A method of inducing cartilage regeneration by transplanting chondrocytes is used as a surgical method for cartilage regeneration.

However, since fibro-cartilage is mainly generated instead of supercartilage required for the actual joint, the micro-fracture technique has a disadvantage in that it does not achieve a satisfactory effect in terms of functionality, and the osteochondral transplantation is There is a disadvantage in that a gap remains between the site and the existing tissue, and in the case of the chondrocyte transplantation, there is a disadvantage in that the patient's pain and economic burden due to multiple surgeries are large. According to these problems, there is still a need to develop a technology capable of regenerating tissues such as cartilage very effectively.

One object of the present invention is to provide a composition for tissue regeneration.

Another object of the present invention is to provide a composition for preventing, improving or treating cartilage disease.

However, the technical task to be achieved by the present invention is not limited to the tasks mentioned above, and other tasks not mentioned will be clearly understood by those of ordinary skill in the art from the following description.

One. Composition for tissue regeneration

One embodiment of the present invention provides a composition for tissue regeneration.

The composition for tissue regeneration of the present invention includes the following ingredients as active ingredients:

1) CK2 (casein kinase 2) protein activity inhibitors or CK2 protein-encoding genes (eg, CSNK2A1, CSNK2A2 and CSNK2B ) expression inhibitors;

2) a cartilage binding peptide or a nucleic acid sequence encoding the same; and

3) Adeno-associated virus (AAV) expression vector.

The present invention provides various types of stem cells by using a cartilage-binding peptide or a nucleic acid sequence encoding the same and an adeno-associated virus (AAV) vector together with the inhibitor of the activity of the CK2 protein or the expression inhibitor of the gene encoding the CK2 protein. can induce and promote effective differentiation into chondrocytes.

The "CK2 protein" of the present invention exists in various eukaryotic cells, and is composed of two molecules of catalytic subunits (α1 and α2) and two molecules of regulatory subunits (β). In the case of CK2 α1, it is encoded by CSNK2A1, in the case of CK2 α2, it is encoded by CSNK2A2, and in the case of CK2 β, it is encoded by CSNK2B. When the gene encoding the α subunit of the CK2 protein is destroyed, the yeast dies, so the CK2 protein is essential for cell growth. For the purpose of the present invention, the CK2 protein may be CK2 α1, CK2 α2 and CK2 β, for example, CK2 β, but is not limited thereto.

The CK2 α1 protein of the present invention may consist of the amino acid sequence shown in SEQ ID NO: 1, the CK2 α2 protein may consist of the amino acid sequence shown in SEQ ID NO: 2, and the CK2 β protein is SEQ ID NO: 3 It may consist of the indicated amino acid sequence, but is not limited thereto.

The amino acid sequence of the present invention can be easily modified by a person skilled in the art according to one or more amino acid substitutions, deletions, insertions, or combinations thereof. Therefore, the activity or expression level of the gene can be reduced even when a protein or polypeptide having high homology with the amino acid sequence shown in SEQ ID NOs: 1 to 3, for example, is 80% or more, 90% or more, or 95% or more. All are included in the protein of the present invention as long as it maintains the technical characteristics of inducing or promoting differentiation of bone or cartilage when inhibited.

The "homology" of the present invention is intended to indicate a degree of similarity to the amino acid sequence of a wild-type protein, and includes a sequence having the same sequence as above with the amino acid sequence of the present invention. . Such homology can be determined by visually comparing two sequences, but can be determined using a bioinformatic algorithm that analyzes the degree of homology by arranging sequences to be compared side by side. The homology between the two amino acid sequences can be expressed as a percentage. Useful automated algorithms are available in the GAP, BESTFIT, FASTA and TFASTA computer software modules of the Wisconsin Genetics Software Package (Genetics Computer Group, Madison, W, USA). Automated sequencing algorithms in the module include Needleman & Wunsch and Pearson & Lipman and Smith & Waterman sequencing algorithms. Algorithms and homology determinations for other useful sequences can be automated in software including FASTP, BLAST, BLAST2, PSIBLAST and CLUSTAL W.

The "protein activity inhibitor" of the present invention may be used without limitation as long as it is a substance capable of inhibiting the kinase activity of CK2 protein, and may include an antibody or fragment thereof capable of specifically binding to the CK2 protein; and any one or more of peptides.

The "antibody" of the present invention may be a polyclonal or monoclonal antibody. In the present invention, the antibody against the CK2 protein is prepared by methods commonly performed in the art, for example, a fusion method (Kohler et al., European Journal of Immunology, 6:511-519 (1976)), a recombinant DNA method. (U.S. Pat. No. 4,816,56) or phage antibody library methods (Clackson et al, Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:58, 1-597 ( 1991)) can be prepared. General procedures for antibody preparation are described in Harlow, E. et al., Using Antibodies: A Laboratory Manual, Cold Spring Harbor Press, New York, 1999; Zola, H., Monoclonal Antibodies: A Manual of Techniques, CRC Press, Inc., Boca Raton, Florida, 1984; and Coligan, CURRENT PROTOCOLS IN IMMUNOLOGY, Wiley/Greene, NY, 1991), which are incorporated herein by reference. For example, the production of moclonal antibody-producing hybridoma cells is accomplished by fusing an immortalized cell line with antibody-producing lymphocytes, and the techniques required for this process are well known to those skilled in the art and can be easily performed. . Polyclonal antibodies can be obtained by injecting the CK2 protein antigen into a suitable animal, collecting antisera from the animal, and then isolating the antibody from the antisera using known affinity techniques.

The "fragment" of the present invention means a fragment having at least an antigen-binding function, and may include Fab, F(ab'), F(ab')2 and Fv, but is not limited thereto.

The "gene expression inhibitor" of the present invention inhibits the transcription or translation process of the gene encoding the CK2 protein, that is, the CK2 gene ( CSNK2A1, CSNK2A2 and CSNK2B). Any substance capable of suppressing the expression of the gene can be used. and, for example, may be any one or more of antisense oligonucleotides and aptamers capable of specifically binding to the gene, but is not limited thereto.

The "antisense oligonucleotide" of the present invention means DNA or RNA or a derivative thereof containing a nucleic acid sequence complementary to a specific mRNA sequence, and binds to a complementary sequence in mRNA to convert mRNA into protein acts as an inhibitor. The antisense sequence of the present invention refers to a DNA or RNA sequence complementary to the CK2 gene and capable of binding to the gene, and translation of the mRNA, translocation into the cytoplasm, maturation or all other overall biological functions. may inhibit essential activity for For example, it may be siRNA, shRNA, etc., and may consist of the nucleotide sequence shown in SEQ ID NO: 4, but is not limited thereto.

The antisense oligonucleotides of the present invention may be modified at one or more bases, sugars or backbone positions to enhance efficacy (De Mesmaeker et al., Curr Opin Struct Biol., 5, 3, 343-55). , 1995). The backbone may be modified with phosphorothioates, phosphotriesters, methyl phosphonates, short chain alkyls, cycloalkyls, short chain heteroatomics, heterocyclic intersugar bonds, and the like. Antisense nucleic acids may also include one or more substituted sugar moieties. Antisense oligonucleotides may include modified bases. Modified bases include hypoxanthine, 6-methyladenine, 5-methylpyrimidine (particularly 5-methylcytosine), 5-hydroxymethylcytosine (HMC), glycosyl HMC, gentobiosyl HMC, 2-aminoadenine, 2 -Thiouracil, 2-thiothymine, 5-bromouracil, 5-hydroxymethyluracil, 8-azaguanine, 7-deazaguanine, N6 (6-aminohexyl) adenine, 2,6-diaminopurine, etc. There is this. In addition, the antisense oligonucleotide of the present invention may be chemically bound to one or more moieties or conjugates that enhance the activity and cell adsorption properties of the antisense oligonucleotide. Cholesterol moiety, cholesteryl moiety, cholic acid, thioether, thiocholesterol, fatty chain, phospholipid, polyamine, polyethylene glycol chain, adamantane acetic acid, palmityl moiety, octadecylamine, hexylamino-carbonyl-oxy fat soluble moieties such as cholesterol moieties, and the like. Oligonucleotides comprising a fat-soluble moiety and methods of preparation are well known in the art (US Pat. Nos. 5,138,045, 5,218,105 and 5,459,255). The modified nucleic acid can increase the stability to nucleases and increase the binding affinity of the antisense oligonucleotide with the target mRNA.

The antisense oligonucleotide of the present invention may be synthesized in vitro by a conventional method and administered in vivo, or the antisense oligonucleotide may be synthesized in vivo. An example of synthesizing antisense oligonucleotides in vitro is using RNA polymerase I. One example of allowing antisense RNA to be synthesized in vivo is to use a vector with the origin of the recognition site (MCS) in the opposite direction to allow the antisense RNA to be transcribed. Such antisense RNA preferably has a translation stop codon in the sequence so that it is not translated into the peptide sequence.

The "aptamer" of the present invention refers to an oligonucleotide molecule having binding activity to a predetermined target molecule. The aptamer can inhibit the activity of a predetermined target molecule by binding to the predetermined target molecule.

The aptamer of the present invention may be RNA, DNA, a modified (modified) oligonucleotide, or a mixture thereof. The aptamer of the present invention may also be in a linear or cyclic form. The length of the aptamer of the present invention is not particularly limited and may be usually 15 to 200 nucleotides, for example, 15 to 100 nucleotides, preferably 18 to 80 nucleotides, more preferably 20 to 60 nucleotides, and most preferably Preferably, it may be 22 to 45 nucleotides. When the total number of nucleotides is small, chemical synthesis, chemical modification, and mass production are easier, economical, and advantageous in that in vivo stability is high and toxicity is low.

The cartilage-binding peptide of the present invention may consist of an amino acid sequence represented by SEQ ID NO: 5 or 6, but is not limited thereto.

The gene encoding the cartilage-binding peptide of the present invention may consist of the nucleotide sequence represented by SEQ ID NO: 7 or 8, but is not limited thereto.

The "adeno-associated virus (AAV)" of the present invention is a kind of parvovirus, and is a non-pathogenic virus that does not induce an immune response even when injected into an individual, and the range of infected host cells is wide, and it is possible to infect not only growing cells but also cells that have stopped growing, and it not only exists in the form of an episome in the host cell, but also inserts the genome of the virus into the chromosome of the host cell. have the ability to do In particular, when the virus is infected with human cells, the genome of the virus tends to be inserted into a specific position on human chromosome 19, which is known as a region where genes responsible for special functions do not exist, so insertion mutations ( Since the possibility of proto-oncogene activation by insertional mutagenesis is very low, it is more advantageous for gene therapy. In addition, the recombinant form of the adeno-associated virus has the advantage that chromosomal integration rarely occurs.

The adeno-associated virus of the present invention may be derived from serotype 2 (AAV2) or 5 (AAV5), and preferably may be AAV derived from serotype 5, but is not limited thereto.

The "expression vector" of the present invention refers to a vector capable of expressing a desired protein in a desired host cell, and refers to a gene construct including essential regulatory elements operably linked to express a gene insert.

The expression vector of the present invention may be an adeno-associated virus (AAV5) expression vector of serotype 5, preferably composed of the nucleotide sequence shown in SEQ ID NO: 9, but is not limited thereto.

The tissue of the present invention may be cartilage tissue, but is not limited thereto.

The "cartilage tissue" of the present invention is a flexible connective tissue found in various parts of the body of animals, including humans, and is a concept including joints between bones, rib cage, ears, nose, bronchial tubes, and intervertebral discs. Unlike bone, it is hard and not rigid, but compared to muscle, it is less flexible and relatively stiff. The cartilage tissue is a collagen fiber, a proteoglycan-rich ground substance, and a chondroblast that produces a large amount of extracellular matrix composed of elastin fiber. It is made up of specialized cells called Nutrients are provided to the cartilage cells trapped in the cartilage tissue through diffusion by pumping action generated by compression of articular cartilage or bending of elastic cartilage. For this reason, the cartilage tissue, unlike other connective tissue, does not include blood vessels, so growth and recovery may be very slow.

The cartilage tissue of the present invention may be at least one selected from the group consisting of elastic cartilage, hyaline cartilage, and fibrocartilage, which can be classified according to the relative content of the three main elements. However, the present invention is not limited thereto.

The site in which the cartilage tissue of the present invention is located is hyaline cartilage (eg, hyaline cartilage; for example, hyaline cartilage located in the joint, nose, larynx or sternum), elastic cartilage (eg, elastic cartilage located in the ear) Or fibrocartilage (eg, intervertebral disc) may be, but is not limited thereto.

The "regeneration" of the present invention generally refers to a product that attempts to restore a part of a body or its function to its original state or restore its function when a part of the body or its function is lost in the present invention. For the purpose of the present invention, the regeneration may mean that the stem cells can be restored to their original state by inducing them to be differentiated into cartilage tissue, but is not limited thereto.

The composition for tissue regeneration of the present invention may be used as a food composition or a pharmaceutical composition.

The pharmaceutical composition of the present invention may be characterized in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the pharmaceutical composition may be characterized in that it is targeted to humans.

The pharmaceutical composition of the present invention is not limited thereto, but each is formulated in the form of oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterile injection solutions according to conventional methods to be used. can The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, dyes, fragrances, etc., for oral administration, and in the case of injections, buffers, preservatives, pain-free agents Agents, solubilizers, isotonic agents, stabilizers, etc. may be mixed and used, and in the case of topical administration, bases, excipients, lubricants, preservatives, etc. may be used. The dosage form of the pharmaceutical composition of the present invention can be prepared in various ways by mixing with a pharmaceutically acceptable carrier as described above. For example, in the case of oral administration, it can be prepared in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and in the case of injections, it can be prepared in the form of unit dose ampoules or multiple doses. there is. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained release formulation, and the like.

Meanwhile, examples of suitable carriers, excipients and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil may be used. In addition, fillers, anti-agglomeration agents, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like may be further included.

The route of administration of the pharmaceutical composition of the present invention is, but not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal. Oral or parenteral administration is preferred.

The parenterals of the present invention include subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

The pharmaceutical composition of the present invention depends on several factors including the activity of the specific compound used, age, weight, general health, sex, formula, administration time, administration route, excretion rate, drug formulation, and the severity of the specific disease to be prevented or treated. The dosage of the pharmaceutical composition may vary depending on the patient's condition, body weight, degree of disease, drug form, administration route and period, but may be appropriately selected by those skilled in the art, and 0.0001 to 50 mg/kg per day Or 0.001 to 50 mg/kg may be administered. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, solutions, gels, syrups, slurries, and suspensions.

The food composition comprising the active ingredient of the present invention may be prepared in the form of various foods, for example, beverages, gums, tea, vitamin complexes, powders, granules, tablets, capsules, sweets, rice cakes, breads, and the like.

When the active ingredient of the present invention is included in the food composition, the amount may be added in a proportion of 0.1 to 50% of the total weight, but is not limited thereto.

When the food composition of the present invention is prepared in the form of a beverage, there is no particular limitation except for including the food composition in the indicated ratio, and it may contain various flavoring agents or natural carbohydrates as additional ingredients, like a conventional beverage. . Specifically, as natural carbohydrates, monosaccharides such as glucose, disaccharides such as fructose, polysaccharides such as sucrose, conventional sugars such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, erythritol, etc. may include. The flavoring agent may be a natural flavoring agent (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agent (saccharin, aspartame, etc.).

The food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, It may further include a pH adjuster, a stabilizer, a preservative, glycerin, alcohol, a carbonation agent used in carbonated beverages, and the like.

The above components of the present invention may be used independently or in combination. The proportion of the additive is not a key element of the present invention, but may be selected from 0.1 to about 50 parts by weight per 100 parts by weight of the food composition of the present invention, but is not limited thereto.

2. Composition for preventing, improving or treating cartilage disease

Another embodiment of the present invention provides a composition for preventing, improving or treating cartilage disease.

The composition for preventing, improving or treating cartilage disease of the present invention comprises the following ingredients as active ingredients:

1) CK2 (casein kinase 2) protein activity inhibitors or CK2 protein-encoding genes (eg, CSNK2A1, CSNK2A2 and CSNK2B ) expression inhibitors;

2) a cartilage binding peptide or a nucleic acid sequence encoding the same; and

3) Adeno-associated virus (AAV) expression vector.

In the composition for preventing, improving or treating cartilage disease of the present invention, CK2 protein and a gene encoding the same, an activity inhibitor of CK2 protein, an expression inhibitor of a gene encoding CK2 protein, a cartilage binding peptide or a nucleic acid sequence encoding the same, adeno -Any virus (AAV) vector, tissue, cartilage, pharmaceutical composition, food composition, etc. are the same as those described in "1. Composition for tissue regeneration", and are omitted to avoid excessive complexity of the specification.

The "cartilage disease" of the present invention refers to any disease that may be caused by damage such as no cartilage or degeneration, for example, arthritis; osteoporosis; osteochondrosis; osteochondritis; incomplete osteoplasty; osteomyelitis; osteophyte; achondroplasia; chondritis; chondroma; chondrosarcoma; herniated disc; Klippel-Pyle Syndrome; osteomyelitis; cystic fibroostitis; and accidents, fractures, wounds, joint damage, autoimmune diseases, diabetes or cancer-induced cartilage diseases, but may be at least one selected from the group consisting of, but not limited to.

The "prevention" of the present invention may include, without limitation, any act of blocking or suppressing or delaying symptoms caused by cartilage disease using the composition of the present invention.

The "improvement" and "treatment" of the present invention may be included without limitation as long as the symptoms caused by cartilage disease are improved or beneficially changed using the composition of the present invention.

The composition of the present invention can induce and promote the differentiation of stem cells into cartilage. Therefore, it can be very effectively used for preventing, improving or treating diseases related to bone or cartilage by promoting cartilage differentiation.

1 is a photograph showing the expression of the peptide after treatment with cartilage-binding peptides (CSP-1, CSP-2) in mouse chondrocytes in Preparation Example 2 of the present invention observed with a fluorescence microscope.
Figure 2 is a western blot of the expression levels of COL2A1 and CK2 β after human bone marrow-derived stem cells (hBMSC) were treated with CSNK2B-specific siRNA, cartilage-binding peptide (CSP-1) and AAV5 vectors in Example 1 of the present invention. Shows the results confirmed by .
Figure 3 shows changes in the expression level of COL2A1 and CK2 β after treatment with CSNK2B-specific siRNA, cartilage-binding peptide (CSP-1) and AAV5 vector in human bone marrow-derived stem cells (hBMSC) in Example 1 of the present invention; shown graphically.
Figure 4 is a CSNK2B-specific siRNA, cartilage-binding peptide (CSP-1) and AAV5 vectors after inducing an inflammatory environment in chondrocytes in Example 2 of the present invention, proteoglycan (proteoglycan) and glycosamino It shows a photograph confirming the degree of decomposition of glycan (glycosaminoglycan).

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited by the following examples.

Example

[Preparation Example 1] Culture of Human Bone Marrow-Derived Mesenchymal Stem Cells (hBMSCs)

All of the following experiments were conducted under the approval of the Institutional Review Board (IRB) (approval number: IRB no. 4-2018-1223). Bone marrow aspirates were obtained from the posterior iliac crest of 6 adult donors. Human bone marrow-derived mesenchymal stem cells (hBMSCs) were selected based on their ability to be adsorbed from the bone marrow puncture to the plastic cell culture flask. The selected hBMSCs were cultured in a culture medium (DMEM-LG (low-glucose Dulbecco's modified Eagle's medium; Gibco) containing 10% FBS (Gibco, Grand Island, NY, USA) and 1% antibiotic-antifungal solution (Gibco)). cultured. Here, by flow cytometry analysis according to a conventional method, only the cells whose characteristics of hBMSCs were confirmed by showing that CD90 and CD105 were positive and CD34 and CD45 were negative were used in the experiment below.

[Preparation Example 2] Confirmation of chondrocyte targeting ability of peptides

Peptides (CSP-1, CSP-2) consisting of the amino acid sequences shown in SEQ ID NOs: 5 and 6 were prepared by requesting Bioneer, and then, rhodamine B was tagged at the N-terminus of each peptide. Mouse chondrocytes, ATDC cells, were inoculated into a 6-well plate at 1 x 10 ⁵ cells/well, and then treated with each peptide at an amount of 2 uM/ml the next day. After 24 hours, the medium was removed, a PBS solution was added, and the result was observed under a fluorescence microscope, and the results are shown in FIG. 1 .

As shown in FIG. 1 , it was confirmed that the peptides (CSP-1, CSP-2) according to the present invention were expressed in mouse chondrocytes.

Through this, it was found that the peptide according to the present invention can specifically bind to chondrocytes.

[Example 1] Confirmation of chondrogenic differentiation ability of bone marrow-derived stem cells

Human bone marrow-derived stem cells (hBMSCs) were inoculated into a 6-well plate at 1 x 10 ⁵ cells/well, and then siCtrl and siCK2b (siRNA) represented by SEQ ID NO: 4 were transfected. After 24 hours, ^{3D micromass culture was performed by making 1 x 10 5} cells/micromass in a 24-well plate. From the next day after micromass culture, the AAV5 vector represented by SEQ ID NO: 8 for each group (1 X 10 ⁹ /ml), treated with the peptide (CSP-1) (2uM/ml) represented by SEQ ID NO: 5. The expression level of COL2A1 protein was confirmed by Western blot on the 7th day of cartilage differentiation, and the results are shown in FIGS.

As shown in FIGS. 2 and 3, in the group treated with the AAV5 vector and the cartilage target peptide after CK2β knockdown, compared to the case where only the AAV5 vector was treated or when the AAV5 vector and the cartilage target peptide (CSP-1) were treated. It was confirmed that the expression level of COL2A1, a protein related to cartilage differentiation, was significantly increased.

[Example 2] Confirmation of protective ability of chondrocytes in inflammatory environment

Chondrocytes, TC28a2, were inoculated into a 6-well plate at 1 x 10 ⁵ cells/well, and then siCtrl and siCK2b (siRNA) represented by SEQ ID NO: 4 were transfected. After 24 hours, 1 x 10 ⁵ cells were inoculated into a 24-well plate to perform high-density culture. From the next day of high-density culture, the AAV5 vector represented by SEQ ID NO: 8 and the peptide (CSP-1) represented by SEQ ID NO: 5 were treated for each group. In order to induce an inflammatory environment, it was treated with LPS 5ug/ml for 5 days. On the 5th day of treatment, the degree of degradation of proteoglycan and glycosaminoglycan was confirmed using safranin O (safranin O) staining, and the results are shown in FIG. 4 .

As shown in Figure 4, after CK2β knockdown, the AAV5 vector and the cartilage target peptide were used together as compared to the control treated with the AAV5 vector only and the AAV5 vector, the AAV5 vector and the cartilage target peptide (CSP-1) after LPS treatment. It was confirmed that the staining remained dark in the treated group. Through this, it was found that when AAV5 vector and cartilage target peptide were treated together with suppression of expression of CK2β protein or a gene encoding it, the effect of inhibiting the degradation of proteoglycans and glycosaminoglycans in chondrocytes in an inflammatory environment was excellent. .

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

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Val Lys Pro His 145 150 155 160 Asn Val Met Ile Asp His Glu His Arg Lys Leu Arg Leu Ile Asp Trp 165 170 175 Gly Leu Ala Glu Phe Tyr His Pro Gly Gln Glu Tyr Asn Val Arg Val 180 185 190 Ala Ser Arg Tyr Phe Lys Gly Pro Glu Leu Leu Val Asp Tyr Gln Met 195 200 205 Tyr Asp Tyr Ser Leu Asp Met Trp Ser Leu Gly Cys Met Leu Ala Ser 210 215 220 Met Ile Phe Arg Lys Glu Pro Phe Phe His Gly His Asp Asn Tyr Asp 225 230 235 240 Gln Leu Val Arg Ile Ala Lys Val Leu Gly Thr Glu Asp Leu Tyr Asp 245 250 255 Tyr Ile Asp Lys Tyr Asn Ile Glu Leu Asp Pro Arg Phe Asn Asp Ile 260 265 270 Leu Gly Arg His Ser Arg Lys Arg Trp Glu Arg Phe Val His Ser Glu 275 280 285 Asn Gln His Leu Val Ser Pro Glu Ala Leu Asp Phe Leu Asp Lys Leu 290 295 300 Leu Arg Tyr Asp His Gln Ser Arg Leu Thr Ala Arg Glu Ala Met Glu 305 310 315 320 His Pro Tyr Phe Tyr Thr Val Val Lys Asp Gln Ala Arg Met Gly Ser 325 330 335 Ser Ser Met Pro Gly Gly Ser Thr Pro Val Ser Ser Ala Asn Met Met 340 345 350 Ser Gly Ile Ser Ser Val Pro Thr Pro Ser Pro Leu Gly Pro Leu Ala 355 360 365 Gly Ser Pro Val Ile Ala Ala Ala Asn Pro Leu Gly Met Pro Val Pro 370 375 380 Ala Ala Ala Gly Ala Gln Gln 385 390 <210> 2 <211> 350 <212> PRT <213> Homo sapiens <400> 2 Met Pro Gly Pro Ala Ala Gly Ser Arg Ala Arg Val Tyr Ala Glu Val 1 5 10 15 Asn Ser Leu Arg Ser Arg Glu Tyr Trp Asp Tyr Glu Ala His Val Pro 20 25 30 Ser Trp Gly Asn Gln Asp Asp Tyr Gln Leu Val Arg Lys Leu Gly Arg 35 40 45 Gly Lys Tyr Ser Glu Val Phe Glu Ala Ile Asn Ile Thr Asn Asn Glu 50 55 60 Arg Val Val Val Lys Ile Leu Lys Pro Val Lys Lys Lys Lys Ile Lys 65 70 75 80 Arg Glu Val Lys Ile Leu Glu Asn Leu Arg Gly Gly Thr Asn Ile Ile 85 90 95 Lys Leu Ile Asp Thr Val Lys Asp Pro Val Ser Lys Thr Pro Ala Leu 100 105 110 Val Phe Glu Tyr Ile Asn Asn Thr Asp Phe Lys Gln Leu Tyr Gln Ile 115 120 125 Leu Thr Asp Phe Asp Ile Arg Phe Tyr Met Tyr Glu Leu Leu Lys Ala 130 135 140 Leu Asp Tyr Cys His Ser Lys Gly Ile Met His Arg Asp Val Lys Pro 145 150 155 160 His Asn Val Met Ile Asp His Gln Gln Lys Lys Leu Arg Leu Ile Asp 165 170 175 Trp Gly Leu Ala Glu Phe Tyr His Pro Ala Gln Glu Tyr Asn Val Arg 180 185 190 Val Ala Ser Arg Tyr Phe Lys Gly Pro Glu Leu Leu Val Asp Tyr Gln 195 200 205 Met Tyr Asp Tyr Ser Leu Asp Met Trp Ser Leu Gly Cys Met Leu Ala 210 215 220 Ser Met Ile Phe Arg Arg Glu Pro Phe Phe His Gly Gln Asp Asn Tyr 225 230 235 240 Asp Gln Leu Val Arg Ile Ala Lys Val Leu Gly Thr Glu Glu Leu Tyr 245 250 255 Gly Tyr Leu Lys Lys Tyr His Ile Asp Leu Asp Pro His Phe Asn Asp 260 265 270 Ile Leu Gly Gln His Ser Arg Lys Arg Trp Glu Asn Phe Ile His Ser 275 280 285 Glu Asn Arg His Leu Val Ser Pro Glu Ala Leu Asp Leu Leu Asp Lys 290 295 300 Leu Leu Arg Tyr Asp His Gln Gln Arg Leu Thr Ala Lys Glu Ala Met 305 310 315 320 Glu His Pro Tyr Phe Tyr Pro Val Val Lys Glu Gln Ser Gln Pro Cys 325 330 335 Ala Asp Asn Ala Val Leu Ser Ser Gly Leu Thr Ala Ala Arg 340 345 350 <210> 3 <211> 215 <212> PRT <213> Homo sapiens <400> 3 Met Ser Ser Ser Glu Glu Val Ser Trp Ile Ser Trp Phe Cys Gly Leu 1 5 10 15 Arg Gly Asn Glu Phe Phe Cys Glu Val Asp Glu Asp Tyr Ile Gln Asp 20 25 30 Lys Phe Asn Leu Thr Gly Leu Asn Glu Gln Val Pro His Tyr Arg Gln 35 40 45 Ala Leu Asp Met Ile Leu Asp Leu Glu Pro Asp Glu Glu Leu Glu Asp 50 55 60 Asn Pro Asn Gln Ser Asp Leu Ile Glu Gln Ala Ala Glu Met Leu Tyr 65 70 75 80 Gly Leu Ile His Ala Arg Tyr Ile Leu Thr Asn Arg Gly Ile Ala Gln 85 90 95 Met Leu Glu Lys Tyr Gln Gln Gly Asp Phe Gly Tyr Cys Pro Arg Val 100 105 110 Tyr Cys Glu Asn Gln Pro Met Leu Pro Ile Gly Leu Ser Asp Ile Pro 115 120 125 Gly Glu Ala Met Val Lys Leu Tyr Cys Pro Lys Cys Met Asp Val Tyr 130 135 140 Thr Pro Lys Ser Ser Arg His His His Thr Asp Gly Ala Tyr Phe Gly 145 150 155 160 Thr Gly Phe Pro His Met Leu Phe Met Val His Pro Glu Tyr Arg Pro 165 170 175 Lys Arg Pro Ala Asn Gln Phe Val Pro Arg Leu Tyr Gly Phe Lys Ile 180 185 190 His Pro Met Ala Tyr Gln Leu Gln Leu Gln Ala Ala Ser Asn Phe Lys 195 200 205 Ser Pro Val Lys Thr Ile Arg 210 215 <210> 4 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> siCK2b <400> 4 gugaagugga ugaagacua 19 <210> 5 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> chondrocyte binding peptide CSP-1 <400> 5 Arg Leu Asp Pro Thr Ser Tyr Leu Arg Thr Phe Trp 1 5 10 <210> 6 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> chondrocyte binding peptide CSP-2 <400> 6 His Asp Ser Gln Leu Glu Ala Leu Ile Lys Phe Met 1 5 10 <210> 7 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> chondrocyte binding peptide CSP-1 <400> 7 cgtcttgatc ctacttctta tttacgtact ttttgg 36 <210> 8 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> chondrocyte binding peptide CSP-2 <400> 8 catgattctc aattagaagc tttaattaaa tttatg 36 <210> 9 <211> 7371 <212> DNA <213> Artificial Sequence <220> <223> AAV5 vector <400> 9 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta 60 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc 120 gccagcgctc gttcaaacct cccgctcaaa atggagaccc tgcgtgctca ctcgggctta 180 aatacccagc gtgaccacat ggtgtcgcaa aatgtcgcaa aacactcacg tgacctctaa 240 tacaggactc tccctaaccc tatgacgtaa ttcacgtcac gactccaccc ctccgcccgc 300 tgtttaaacg cccgggctgt agttaatgat taacccgcca tgctacttat ctacgtagtt 360 tattgattaa caagcattaa aggggtcggg taaggtatcg ggttccgata ggtctggtgg 420 ttctgtattc cccggtgctg tccggggcaa agtccacaaa ctgggggtcg ttgtagttgt 480 ttgtgtactg gatctctggg ttccacctct tggagttttc cttcttgagc tcccactcca 540 tctccacggt gacctgcccg gtgctgtact gggtgatgaa gctgctgacg ggcacgtccg 600 agaagctggt gatatttccg ggcacaggcg tgttcttgat gagcatcatg ggcggtgggt 660 gtttgagtcc gaatccgccc atggccggag aggggtgaaa gtgcgccccc gtctctggga 720 tcttggccca gatgggtcct tggaggtaca cgtccctctc catccacacg ctgccgggca 780 cgatttcctg gaggttgtac gtgccggtcg cgggggcagt ggtggagctc tggttgttgg 840 tggccatctg cccgccgacg ttgtacgcca cgcggttcac cggctgcgtc tcgctctcgc 900 tggtgatgag catgttgccc tcgaggtacg tggcggtggt gcccgggttc gccggctggc 960 tgttgaagat catagtgttc tccagggcat aggtgttgct gccctggagg ttgttggtca 1020 tgccgttcgg ctgcgggggc acctggtaac tcgcgccctc gagctccatc ctattggtcg 1080 tggcgaaggc gctgacactg gcgcggttga ccccggagcc caggttccag ccctgggttc 1140 ggcccatggg ccccgggaac cagtttttgt aggtgttggc gtatctcccg gccaggttct 1200 tgttgaactg gactccgcca gtgttatttg tgctcacgaa gcggtacaag tactggtcca 1260 ccagcgggtt ggccagcttg aacaggttct gactgggagc gaagctggag tggaagggca 1320 cctcctcaaa gttgtaggta aactcaaagt tgttgcccgt tctcagcatc ttgctgggaa 1380 agtactctag gcagaagaag ctgctcctct cggtgggatt ttctgtgttg tcgcggttca 1440 gcgtcgcgta accgtactgc ggcagcgtaa agacctgcgg agggaaggcc ggcaggcatc 1500 cctcggtccc gttgccgacg acgtagggca gctggtagtc gtcgtccgta aacacttgga 1560 cggtggaggt gaggttgttg gcgatggtgg tggtggagtc ctgcaccgtg acctctttga 1620 cttgaatgtt gaagattttg actctgaggg accggggtct gaagccccag tagttgttga 1680 tgagtctttg ccagtctcgg gggctccagt ggctgtggaa gcggttaaag tcaaagtacc 1740 cccagggggt gctgtatcca aagtaggcgt tggcgttgct tccgtcgacg gagccgcttt 1800 tgatctctcg gtactggtgg ttgttgtagc tgggcagcac ccaggttcgg gtggacttgg 1860 tgacgactct gtcccccatc cacgtggaat cgcaatgcca atctcccgag gcattgccca 1920 ctccatcggc accttggtta ttgtcgccca atgggccgcc acctcccgca gacattgtat 1980 cagctcccaa acttgaggct ggttgggctg ggatttgcag ctgctgggat ccgctgggtc 2040 cagcttcggc gtctgacgag gtggaaggct tggagtcctc ttcggtccga gccttctttc 2100 tttttggaaa gtggtcgtct atccgctttc cggtaggggc cgtcttagca ccctcttcaa 2160 ccaggccaaa aggttcgaga acccttttct tggcctgaaa gactgccttt ccgaggtttc 2220 ccccgaagga tgtgtcgtcg gcgagcttct cctgaaactc ggcgtccgcg tggttgtact 2280 tgaggtaggg gttgtctccc gcctcaagct gctcgttgta cgagatgtcg tgctctcgcg 2340 cgacctcgtc tgccctgttg acaggctctc ctcgatcgag accgtttccg ggtccgagat 2400 agttataacc aggcagcaca agaccacggg cttgatcttg atgctgctga ttgggttttg 2460 gtttcggtgg gcccgcttca aggcccaaaa actcgcgaag accttcacca acttcttcca 2520 accaatctgg agggtgatca acaaaagaca tacctgattt aaatcattta ttgttcaaag 2580 atgcagtcat ccaaatccac attgaccaga tcgcaggcag tgcaagcgtc tggcaccttt 2640 cccatgatat gatgaatgta gcacagtttc tgatacgcct ttttgacgac agaaacgggt 2700 tgagattctg acacgggaaa gcactctaaa cagtctttct gtccgtgagt gaagcagata 2760 tttgaattct gattcattct ctcgcattgt ctgcagggaa acagcatcag attcatgccc 2820 acgtgacgag aacatttgtt ttggtacctg tctgcgtagt tgatcgaagc ttccgcgtct 2880 gacgtcgatg gctgcgcaac tgactcgcgc acccgtttgg gctcacttat atctgcgtca 2940 ctgggggcgg gtcttttctt ggctccaccc tttttgacgt agaattcatg ctccacctca 3000 accacgtgat cctttgccca ccggaaaaag tctttgactt cctgcttggt gaccttccca 3060 aagtcatgat ccagacggcg ggtgagttca aatttgaaca tccggtcttg caacggctgc 3120 tggtgttcga aggtcgttga gttcccgtca atcacggcgc acatgttggt gttggaggtg 3180 acgatcacgg gagtcgggtc tatctgggcc gaggacttgc atttctggtc cacgcgcacc 3240 ttgcttcctc cgagaatggc tttggccgac tccacgacct tggcggtcat cttcccctcc 3300 tcccaccaga tcaccatctt gtcgacacag tcgttgaagg gaaagttctc attggtccag 3360 tttacgcacc cgtagaaggg cacagtgtgg gctatggcct ccgcgatgtt ggtcttcccg 3420 gtagttgcag gcccaaacag ccagatggtg ttcctcttgc cgaacttttt cgtggcccat 3480 cccagaaaga cggaagccgc atattgggga tcgtacccgt ttagttccaa aattttataa 3540 atccgattgc tggaaatgtc ctccacgggc tgctggccca ccaggtagtc gggggcggtt 3600 ttagtcaggc tcataatctt tcccgcattg tccaaggcag ccttgatttg ggaccgcgag 3660 ttggaggccg cattgaagga gatgtatgag gcctggtcct cctggatcca ctgcttctcc 3720 gaggtaatcc ccttgtccac gagccacccg accagctcca tgtacctggc tgaagttttt 3780 gatctgatca ccggcgcatc agaattggga ttctgattct ctttgttctg ctcctgcgtc 3840 tgcgacacgt gcgtcagatg ctgcgccacc aaccgtttac gctccgtgag attcaaacag 3900 gcgcttaaat actgttccat attagtccac gcccactgga gctcaggctg ggttttgggg 3960 agcaagtaat tggggatgta gcactcatcc accaccttgt tcccgcctcc ggcgccattt 4020 ctggtctttg tgaccgcgaa ccagtttggc aaagtcggct cgatcccgcg gtaaattctc 4080 tgaatcagtt tttcgcgaat ctgactcagg aaacgtccca aaaccatgga tttcaccccg 4140 gtggtttcca cgagcacgtg catgtggaag tagctctctc ccttctcaaa ttgcacaaag 4200 aaaagggcct ccggggcctt actcacacgg cgccattccg tcagaaagtc gcgctgcagc 4260 ttctcggcca cggtcagggg tgcctgctca atcagattca gatccatgtc agaatctggc 4320 ggcaactccc attccttctc ggccacccag ttcacaaagc tgtcagaaat gccgggcaga 4380 tgctcgtcaa ggtcgctggg gaccttaatc acaatctcgt aaaaccccgg cgtggcggct 4440 gcgcgttcaa acctcccgct tcaaaatgga gaccctgcgt gctcactcgg gcttaaatac 4500 ccagcgtgac cacatggtgt cgcaaaatgt cgcaaaacac tcacgtgacc tctaatacag 4560 gacctctagg ggtaccgagc tcgaatagat cagaagttcc tatactttct agagaatagg 4620 aacttcggaa taggaacttc tgatcttccg ggggatccac tagttctaga gcggccggcg 4680 cggggcgtta acgatatcgg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg 4740 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 4800 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 4860 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 4920 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 4980 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 5040 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 5100 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 5160 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 5220 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 5280 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 5340 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 5400 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 5460 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 5520 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 5580 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5640 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5700 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5760 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5820 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5880 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5940 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6000 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6060 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 6120 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6180 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6240 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6300 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6360 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6420 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6480 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6540 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6600 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6660 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6720 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6780 ttatgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6840 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc 6900 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 6960 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 7020 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 7080 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 7140 tttgacgttg gagtccacgt tcttaatagt ggactcttgt tccaaactgg aacaacactc 7200 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 7260 ttaaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct 7320 tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc g 7371

Claims (20)

1) CK2 (casein kinase 2) protein activity inhibitor or CK2 protein-encoding gene expression inhibitor;
2) a cartilage binding peptide or a nucleic acid sequence encoding the same; and
3) A pharmaceutical composition for tissue regeneration comprising an adeno-associated virus (AAV) expression vector as an active ingredient. The method of claim 1,
The tissue is a cartilage tissue, a pharmaceutical composition for tissue regeneration. 3. The method of claim 2,
The cartilage tissue is at least one selected from the group consisting of hyaline cartilage (Hyaline cartilage), elastic cartilage (Elastic cartilage) and fibrocartilage (Fibrocartilage), a pharmaceutical composition for tissue regeneration. The method of claim 1,
The CK2 protein activity inhibitor may include an antibody or fragment thereof that specifically binds to the CK2 protein; and peptides; One or more of those, a pharmaceutical composition for tissue regeneration. The method of claim 1,
The CK2 protein-encoding gene expression inhibitor is one or more of an antisense oligonucleotide and an aptamer capable of specifically binding to the CK2 gene, a pharmaceutical composition for tissue regeneration. The method of claim 1,
The CK2 is at least one selected from the group consisting of CK2 α 1, CK2 α 2 and CK2 β, a pharmaceutical composition for tissue regeneration. The method of claim 1,
The cartilage-binding peptide is composed of the amino acid sequence represented by SEQ ID NO: 5 or 6, a pharmaceutical composition for tissue regeneration. The method of claim 1,
The adeno-associated virus (AAV) expression vector is a serotype 5 adeno-associated virus (AAV5) vector, a pharmaceutical composition for tissue regeneration. 1) CK2 (casein kinase 2) protein activity inhibitor or CK2 protein-encoding gene expression inhibitor;
2) a cartilage binding peptide or a nucleic acid sequence encoding the same; and
3) A pharmaceutical composition for preventing or treating cartilage disease, comprising an adeno-associated virus (AAV) expression vector as an active ingredient. 10. The method of claim 9,
The cartilage disease is arthritis; osteoporosis; osteochondrosis; osteochondritis; incomplete osteoplasty; osteomyelitis; osteophyte; achondroplasia; chondritis; chondroma; chondrosarcoma; herniated disc; Klippel-Pyle Syndrome; osteomyelitis; cystic fibroostitis; And accidents, fractures, wounds, joint damage, autoimmune disease, diabetes, or at least one selected from the group consisting of cartilage disease caused by cancer, a pharmaceutical composition for the prevention or treatment of cartilage disease. 10. The method of claim 9,
The CK2 is at least one selected from the group consisting of CK2 α 1, CK2 α 2 and CK2 β, a pharmaceutical composition for preventing or treating cartilage disease. 10. The method of claim 9,
The cartilage-binding peptide is composed of the amino acid sequence represented by SEQ ID NO: 5 or 6, a pharmaceutical composition for the prevention or treatment of cartilage disease. 10. The method of claim 9,
The adeno-associated virus (AAV) expression vector is a serotype 5 adeno-associated virus (AAV5) vector, a pharmaceutical composition for preventing or treating cartilage disease. 1) CK2 (casein kinase 2) protein activity inhibitor or CK2 protein-encoding gene expression inhibitor;
2) a cartilage binding peptide or a nucleic acid sequence encoding the same; and
3) A food composition for tissue regeneration comprising an adeno-associated virus (AAV) expression vector as an active ingredient. 15. The method of claim 14,
The cartilage-binding peptide is composed of an amino acid sequence represented by SEQ ID NO: 5 or 6, a food composition for tissue regeneration. 15. The method of claim 14,
The adeno-associated virus (AAV) expression vector is a serotype 5 adeno-associated virus (AAV5) vector, a food composition for tissue regeneration. 1) CK2 (casein kinase 2) protein activity inhibitor or CK2 protein-encoding gene expression inhibitor;
2) a cartilage binding peptide or a nucleic acid sequence encoding the same; and
3) A food composition for preventing or improving cartilage disease, comprising an adeno-associated virus (AAV) expression vector as an active ingredient. 18. The method of claim 17,
The cartilage disease is arthritis; osteoporosis; osteochondrosis; osteochondritis; incomplete osteoplasty; osteomyelitis; osteophyte; achondroplasia; chondritis; chondroma; chondrosarcoma; herniated disc; Klippel-Pyle Syndrome; osteomyelitis; cystic fibroostitis; And accidents, fractures, wounds, joint damage, autoimmune diseases, diabetes, or at least one selected from the group consisting of cartilage diseases due to cancer, the prevention or improvement of cartilage disease food composition. 18. The method of claim 17,
The cartilage-binding peptide is composed of an amino acid sequence represented by SEQ ID NO: 5 or 6, a food composition for preventing or improving cartilage disease. 18. The method of claim 17,
The adeno-associated virus (AAV) expression vector is a serotype 5 adeno-associated virus (AAV5) vector, a food composition for preventing or improving cartilage disease.

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