Composition for inhibiting activity or differentiation of m1 macrophages and inducing differentiation into m2 macrophages, comprising mesenchymal stem cells secreting activin a

Abstract

The present invention relates to: a composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages, comprising mesenchymal stem cells secreting activin A; and a method for preventing or treating rheumatoid arthritis by using same.

Description

Composition for inhibiting the activation or differentiation of M1 macrophages and inducing differentiation into M2 macrophages, including mesenchymal stem cells secreting activin A

[0001] The present invention relates to a composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages, including mesenchymal stem cells that secrete activin A, and a method for preventing or treating rheumatoid arthritis using the same.

[0002]

[0003] Mesenchymal stem cells (MSCs) possess self-renewal and proliferation capabilities, and have relatively low MHC class I antigen expression and extremely low MHC class II antigen expression compared to other somatic cells, making them attractive as allogeneic cell therapy. Furthermore, they can be readily expanded in vitro and differentiate into a variety of tissue cells, including adipocytes, osteocytes, chondrocytes, and muscle cells. Therefore, existing stem cell research has focused on treating intractable diseases by utilizing their differentiation capabilities. Recent studies have confirmed the immunomodulatory effects of adult stem cells, which have been demonstrated to be mediated through intercellular contact and the secretion of soluble factors.

[0004] Among them, human umbilical cord blood-derived mesenchymal stem cells secrete high amounts of various factors related to immune or inflammatory responses in cytokine secretion and Gene Ontology (GO) analysis. Among them, Activin A is a TGF-β cytokine known to be involved in various physiological functions such as inflammation, immune regulation, cell growth, and differentiation, but its function or mechanism of action is not yet well known.

[0005]

[0006] Meanwhile, rheumatoid arthritis is a multi-inflammatory joint disease caused by many factors, including immunological abnormalities such as systemic inflammation, activation of T lymphocytes, abnormalities in the cytokine system, abnormal differentiation of macrophages, and abnormal secretion of inflammatory factors, as well as physiological factors and biochemical defects. To treat rheumatoid arthritis, conventional synthetic DMARDs (csDMARDs) based on MTX are used as first-line treatment. However, for patients who do not respond sufficiently to csDMARDs, biological DMARDs (DMARDs) are used. Subsequently, depending on the stage or severity of rheumatoid arthritis, combination therapy with other drugs, including csDMARDs, or targeted synthetic DMARDs (tsDMARDs) are used. However, long-term use of these drugs can cause side effects such as indigestion, osteoporosis, gastrointestinal symptoms, liver toxicity, and bone marrow suppression, and there is a high possibility of relapse when the medication is discontinued. Therefore, there is still a need for safe and effective treatments for long-term use, and the development of more fundamental and groundbreaking treatments is urgently needed.

[0007]

[0008] Against this backdrop, the inventors of the present invention have completed the present invention by confirming that a composition containing activin A or mesenchymal stem cells secreting the composition can effectively suppress pro-inflammatory immune responses in the body and promote anti-inflammatory immune responses, thereby enabling a multifaceted response to abnormal immune responses, and can fundamentally treat rheumatoid arthritis by minimizing the suppression of abnormal immune responses, by inhibiting the activation or differentiation of M1 macrophages and inducing differentiation into M2 macrophages, compared to existing therapeutic agents.

[0009]

[0010] One object of the present invention is to provide a composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages, the composition comprising activin A or a mesenchymal stem cell secreting the composition as an active ingredient.

[0011] Another object of the present invention is to provide a composition comprising activin A, or a composition comprising mesenchymal stem cells secreting the composition as an active ingredient, for use in inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages.

[0012] Another object of the present invention is to provide a method for inhibiting the activation or differentiation of M1 macrophages and inducing differentiation into M2 macrophages, comprising a step of treating a composition comprising activin A or mesenchymal stem cells secreting the composition.

[0013] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating rheumatoid arthritis, comprising a composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages.

[0014] Another object of the present invention is to provide a method for preventing or treating rheumatoid arthritis, comprising a step of administering the pharmaceutical composition to a subject.

[0015] Another object of the present invention is to provide a food composition for preventing or improving rheumatoid arthritis, comprising a composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages.

[0016] Another object of the present invention is to provide a composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages for use in preventing, improving or treating rheumatoid arthritis.

[0017] Another object of the present invention is to provide a composition for inducing and boosting differentiation into M2 macrophages in an M2 macrophage differentiation environment, comprising activin A as an active ingredient.

[0018] Another object of the present invention is to provide a composition comprising activin A as an active ingredient for use in inducing and boosting differentiation into M2 macrophages in an M2 macrophage differentiation environment.

[0019] Another object of the present invention is to provide a method for inducing and boosting differentiation into M2 macrophages in an M2 macrophage differentiation environment, comprising a step of treating a composition containing activin A as an active ingredient.

[0020]

[0021] Unlike existing drugs that simply relieve pain and control only one target factor, the composition of the present invention suppresses the activation or differentiation of M1 macrophages containing mesenchymal stem cells that secrete activin A and induces differentiation into M2 macrophages, thereby inhibiting pro-inflammatory immune responses in the body and promoting anti-inflammatory immune responses, thereby enabling a multifaceted response to abnormal immune responses, and by minimizing the suppression of abnormal immune responses, it can be expected to be a fundamental therapeutic agent for the treatment of rheumatoid arthritis.

[0022]

[0023] Figure 1 shows the results of profiling the secreted factors of each lot of umbilical cord blood-derived mesenchymal stem cells using an antibody array (A, B), and measuring the amount of Activin A secreted using an Activin A Quantikine ELISA kit (B).

[0024] Figure 2 shows the results of measuring the amount of activin A secretion following inhibition of activin A gene expression of umbilical cord blood-derived mesenchymal stem cells using siRNA (small interfering RNA) (A), measuring the amount of TNF-a secretion using a TNF-a Quantikine ELISA kit after co-culturing M1 macrophages and umbilical cord blood-derived mesenchymal stem cells (siCTL or siActivin A) (B), and measuring the amount of IL-10 secretion using an IL-10 Quantikine ELISA kit after co-culturing M0 macrophages and umbilical cord blood-derived mesenchymal stem cells (siCTL or siActivin A) (C).

[0025] Figure 3 shows the results of confirming the amount of IL-10 secreted according to the concentration of recombinant protein activin A in an environment inducing M2 macrophage differentiation using the IL-10 Quantikine ELISA kit.

[0026] Figure 4 shows the results of confirming lymphocyte proliferation and activity according to the presence or absence of co-culture with umbilical cord blood-derived mesenchymal stem cells.

[0027] Figure 5 shows the results of confirming the efficacy of umbilical cord blood-derived mesenchymal stem cells in a rheumatoid arthritis mouse model by examining the arthritis index and staining foot tissue with Hematoxylin and Eosin (H&E) and Safranin O.

[0028] Figure 6 shows the results of evaluating the efficacy of Activin A in a rheumatoid arthritis mouse model.

[0029] Figure 7 is a schematic diagram illustrating a method for culturing macrophages and measuring TNF-a and IL-10 secretion.

[0030] Figure 8 shows the results of confirming the critical point of a composition containing Activin A through TNF-a secretion inhibition efficacy.

[0031] Figure 9 shows the results of a test to confirm the effectiveness of a composition containing MSC or Activin A in inducing IL-10 secretion.

[0032]

[0033] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. In other words, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions described below.

[0034]

[0035] One aspect of the present invention is a composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages, comprising a composition comprising activin A or a mesenchymal stem cell secreting the composition as an active ingredient.

[0036] In one specific example, the composition comprising activin A or the cord blood-derived mesenchymal stem cells secreting the composition are characterized in that they regulate immunity by suppressing the activity or differentiation of M1 macrophages while simultaneously inducing differentiation of M0 macrophages into M2 macrophages.

[0037] A composition for inhibiting the activation or differentiation of M1 macrophages and inducing the differentiation of M0 macrophages into M2 macrophages according to the above specific example, wherein the composition comprising activin A is characterized by comprising activin A, THBS1, GRN, and SPARC.

[0038] A composition according to any one of the preceding specific examples, wherein the composition comprising activin A is further characterized in that it comprises at least one selected from the group consisting of VEGF, EDA, MIF, TIMP-2, and TGF-b1.

[0039] A composition according to any one of the preceding specific examples, wherein the composition comprising activin A or mesenchymal stem cells secreting the composition are co-cultured with M0 macrophages at a ratio of 0.01 to 2:1.

[0040] A composition according to any one of the preceding specific examples, wherein the composition comprising activin A is characterized in that the activin A is 0.1 pg / ml or more and less than 20,000 pg / ml.

[0041] A composition according to any one of the preceding specific examples, wherein the composition comprising activin A is characterized in that the activin A is 1,000 pg / ml or more and less than 20,000 pg / ml.

[0042] A composition according to any one of the preceding specific examples, wherein the composition comprising activin A is characterized in that the activin A is 1,000 pg / ml or more and less than 10,000 pg / ml.

[0043] A composition according to any one of the preceding specific examples, wherein the composition comprising activin A is characterized in that the activin A content is 1,000 pg / ml or more and less than 5,000 pg / ml.

[0044] A composition according to any one of the preceding specific examples, wherein the activin A is characterized in that it boosts the induction and differentiation of M0 macrophages into M2 macrophages in an M2 macrophage differentiation environment.

[0045] A composition according to any one of the preceding specific examples, wherein the M2 macrophage differentiation environment is characterized in that it is an environment containing IL-4 and IL-13, or an environment in which rheumatoid arthritis has developed.

[0046] A composition according to any one of the preceding specific examples, wherein the mesenchymal stem cells are mesenchymal stem cells derived from umbilical cord blood.

[0047]

[0048] Another aspect of the present invention is a method for inhibiting the activation or differentiation of M1 macrophages and inducing differentiation into M2 macrophages, comprising a step of treating a composition comprising activin A or mesenchymal stem cells secreting the composition.

[0049]

[0050] Another aspect of the present invention is a pharmaceutical composition for preventing or treating rheumatoid arthritis, comprising a composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages.

[0051] In one specific embodiment, the composition is characterized in that it further comprises a pharmaceutically acceptable carrier.

[0052] A pharmaceutical composition according to any one of the preceding specific examples, wherein the pharmaceutically acceptable carrier further comprises at least one selected from the group consisting of a preservative, a solubilizer, a stabilizer, an excipient, a lubricant, a preservative, and an additive.

[0053] A pharmaceutical composition according to any one of the preceding specific examples, wherein the composition comprising activin A or the cord blood-derived mesenchymal stem cells secreting the composition suppresses differentiation and activity of M1 macrophages and simultaneously promotes M2 differentiation, thereby regulating immunity.

[0054]

[0055] Another aspect of the present invention is a method for preventing or treating rheumatoid arthritis, comprising administering the pharmaceutical composition to a subject.

[0056]

[0057] Another aspect of the present invention is a composition for inducing and boosting differentiation of M0 macrophages into M2 macrophages in an M2 macrophage differentiation environment, comprising activin A as an active ingredient.

[0058] In one specific example, the M2 macrophage differentiation environment is characterized as an environment containing IL-4 and IL-13, or an environment in which rheumatoid arthritis has developed.

[0059]

[0060] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. In other words, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions described below.

[0061] Furthermore, those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments of the invention described herein. Furthermore, such equivalents are intended to be encompassed by the present invention.

[0062] Additionally, numerous papers and patents are referenced and cited throughout this specification. The disclosures of these cited papers and patents are incorporated herein by reference in their entirety to provide a clearer understanding of the state of the art and the scope of the present invention.

[0063]

[0064] In the present invention, the term "Activin A" is a member of the TGF-β superfamily and is a substance having a wide range of biological activities such as mesoderm induction, neural cell differentiation, bone remodeling, and hematopoiesis, and may be used interchangeably with INHBA (Inhibin, beta A).

[0065] The above term, "composition comprising activin A" means a composition comprising factors secreted from mesenchymal stem cells cultured in a Starvation Medium excluding serum and cytokines necessary for cell growth. Specifically, the composition comprising activin A may be a composition comprising activin A, THBS1 (Thrombospondin 1), GRN, and SPARC, and may further be a composition comprising factors secreted from mesenchymal stem cells cultured in the above stem cells, such as VEGF, EDA, MIF, TIMP-2, and / or TGF-b1, or may be a composition comprising VEGF, EDA, IGFBP7, MDK, MIF, CLU, TIMP-2, and / or TGF-b1, but may include all factors secreted from stem cells when culturing mesenchymal stem cells without limitation.

[0066] The above starvation medium is a medium excluding serum and cytokines required for stem cell growth. Here, the serum required for cell growth may include fetal bovine serum (FBS) and basal serum (BCS), and the cytokines may include basic Fibroblast growth factor (bFGF) and Fibroblast growth factor 2 (FGF2), but the serum or cytokines required for cell growth may be included without limitation.

[0067] The above term, “mesenchymal stem cells secreting the composition” means mesenchymal stem cells secreting a composition containing activin A as described above, wherein the composition containing activin A is as described above.

[0068] In the present invention, "stem cells" are cells capable of differentiating into various tissues, i.e., undifferentiated cells. The stem cells may be of human or animal origin, and may be derived from umbilical cord, umbilical cord blood, bone marrow, fat, muscle, nerves, skin, amniotic membrane, or placenta. For example, they may be mesenchymal stem cells.

[0069] "Mesenchymal stem cells" refer to a heterogeneous population of stem cells that have the ability to self-renew and differentiate into the mesodermal lineage and other embryonic lineages, such as endoderm and ectoderm. Mesenchymal stem cells can be used synonymously with multipotent undifferentiated cells, and are adult stem cells that have the ability to differentiate into various mesodermal cells, including adipocytes, osteoblasts, chondrocytes, cardiac cells, or muscle cells, or ectodermal cells, such as neural cells.

[0070] Specifically, the stem cells of the present invention may be mesenchymal stem cells derived from umbilical cord blood.

[0071]

[0072] The composition comprising the above activin A or the mesenchymal stem cells secreting the same may have a novel use of inhibiting the activity or differentiation of M1 macrophages and promoting the differentiation of M0 macrophages into M2 macrophages. Specifically, the composition comprising the above activin A or the mesenchymal stem cells derived from umbilical cord blood secreting the composition may be characterized by regulating immunity by inhibiting the activity or differentiation of M1 macrophages and promoting the differentiation of M0 macrophages into M2 macrophages.

[0073] In the present invention, the term macrophage is a cell that plays a central role in the pathophysiology of rheumatoid arthritis (RA), and plays a role in causing inflammation and tissue destruction by secreting inflammatory cytokines and tissue-degrading enzymes.

[0074] "M0 macrophages" are undifferentiated, or not yet differentiated, macrophages that differentiate or polarize into anti-inflammatory M2 macrophages and pro-inflammatory M1 macrophages. Of these, M1 macrophages are generally activated by IFN-γ or lipopolysaccharide (LPS), produce pro-inflammatory cytokines such as TNF-α, IL-1b, and IL-6, and initiate immune responses by eliminating microorganisms through phagocytosis. On the other hand, M2 macrophages are differentiated and activated by specific cytokines such as IL-4 and IL-13, and secrete several anti-inflammatory cytokines such as TGF-β and IL-10, which induce tissue remodeling in rheumatoid arthritis and promote wound healing and tissue repair (Aging (Albany NY). 2019 Jul 29;11(14):5258-5275., Biochemical Pharmacology 165 (2019) 152-169).

[0075] Whether the above-mentioned M1 macrophages are activated or differentiated, or whether differentiation is inhibited, can be confirmed by measuring the amount of production or secretion of TNF-a, an inflammatory cytokine, and whether M0 macrophages are differentiated into M2 macrophages or whether differentiation is promoted can be confirmed by measuring the amount of secretion or production of IL-10 secreted during differentiation of M2 macrophages.

[0076] As used herein, the term "differentiation" refers to the phenomenon in which cells become specialized in structure or function during their growth through division and proliferation, i.e., the change in form or function of cells, tissues, etc. of living organisms to perform their respective assigned tasks. For example, differentiation refers to the development of qualitative differences between initially homogeneous parts of a biological system during ontogeny, or the resulting division into qualitatively distinguishable subsystems.

[0077] The above terms, differentiation inhibition, refer to slowing and / or reducing the growth of cells by dividing and proliferating below their normal differentiation level, and differentiation promotion refer to enabling cells to differentiate faster and / or to a greater extent than their normal differentiation level.

[0078] The above "differentiated cell" refers to any cell that is in the process of differentiating into a somatic cell lineage or has been finally differentiated. In other words, it refers to a cell that constitutes an adult and has limited differentiation and self-reproduction capabilities.

[0079] The stem cell culture medium of the present invention can be any medium known for stem cell culture, and specifically, KSB-3 medium can be used.

[0080]

[0081] The present invention confirmed that a composition containing the above activin A is effective in inhibiting the activity or differentiation of M1 macrophages and promoting the differentiation of M0 macrophages into M2 macrophages at a certain content.

[0082] Specifically, the composition comprising the activin A may have an amount of 0.1 pg / ml or more but less than 20,000 pg / ml, 1 pg / ml or more but less than 20,000 pg / ml, 10 pg / ml or more but less than 20,000 pg / ml, 50 pg / ml or more but less than 20,000 pg / ml, 100 pg / ml or more but less than 20,000 pg / ml, 1,000 pg / ml or more but less than 20,000 pg / ml, and more specifically 0.1 pg / ml or more but less than 10,000 pg / ml, 10 pg / ml or more but less than 10,000 pg / ml, 50 pg / ml or more but less than 10,000 pg / ml, 100 pg / ml or more but less than 10,000 pg / ml, 1,000 pg / ml or more. It may be less than or equal to 10,000 pg / ml, and more specifically, it may be more than or equal to 0.1 pg / ml and less than or equal to 5,000 pg / ml, more than or equal to 1 pg / ml and less than or equal to 5,000 pg / ml, more than or equal to 10 pg / ml and less than or equal to 5,000 pg / ml, more than or equal to 100 pg / ml and less than or equal to 5,000 pg / ml, or more than or equal to 1,000 pg / ml and less than or equal to 5,000 pg / ml.

[0083] The composition containing the above activin A or the mesenchymal stem cells secreting the above composition can be mixed with M0 macrophages at a ratio of 0.01 to 2:1 during co-culture. Specifically, a mixing ratio of 0.1 to 2:1 or 1 to 2:1 can also be included.

[0084] The composition containing the above activin A or the cord blood-derived mesenchymal stem cells secreting the above composition are characterized by regulating immunity by inhibiting the activation or differentiation into M1 macrophages and promoting the differentiation of M0 macrophages into M2 macrophages.

[0085] Here, the above activin A may be characterized by boosting the induction and differentiation of M2 macrophages in an M2 macrophage differentiation environment.

[0086] The above term boosting means that activin A itself cannot induce or promote differentiation of M0 macrophages into M2 macrophages, but can only induce and promote differentiation into M2 macrophages in an M2 macrophage differentiation environment.

[0087] The above M2 macrophage differentiation environment may include an environment containing IL-4 and IL-13, or an environment in which rheumatoid arthritis has developed. More specifically, the environment in which rheumatoid arthritis has developed refers to an environment in which inflammatory cells are activated due to increased levels of inflammatory cytokines TNF-α, IL-1, IL-6, and / or IL-17.

[0088] In a specific embodiment of the present invention, it was confirmed that a composition containing activin A or mesenchymal stem cells secreting the same significantly suppresses the amount of TNF-a secretion, effectively inhibits the activity or differentiation into M1 macrophages, and increases the amount of IL-10 secretion, effectively promoting the differentiation of M0 macrophages into M2 macrophages.

[0089] In particular, it was confirmed that a composition containing activin A significantly suppressed the amount of TNF-a secretion in a composition containing all other factors except activin A, thereby effectively suppressing the activation or differentiation into M1 macrophages, and increased the amount of IL-10 secretion, thereby effectively promoting the differentiation of M0 macrophages into M2 macrophages (compare Figs. 2 and 3). In particular, it was confirmed that when the content of the composition containing activin A was 0.1 pg / ml or more and less than 20,000 pg / ml, and particularly 0.1 pg / ml or more and 10,000 pg / ml or less, the amount of TNF-a secretion was significantly suppressed, thereby significantly suppressing the activation or differentiation into M1 macrophages. On the other hand, when the content was 20,000 pg / ml or more, the amount of TNF-a secretion increased rapidly, and when compared to PC (M1), it also increased rapidly, confirming that inhibition of the activation or differentiation into M1 macrophages was not observed from that concentration.

[0090] In addition, it was confirmed that when the composition containing the activin A or the mesenchymal stem cells secreting the composition were co-cultured with M0 macrophages at a ratio of 0.01 to 2: 1, the secretion amount of IL-10 increased, thereby inducing and promoting differentiation of M0 macrophages into M2 macrophages. When this was quantified, when the content of the composition containing activin A was 0.1 pg / ml or more and less than 20,000 pg / ml, more effectively 0.1 pg / ml or more and 10,000 pg / ml or less, the secretion amount of IL-10 rapidly increased, and when it exceeded the value, the secretion amount of IL-10 decreased, thereby promoting differentiation of M0 macrophages into M2 macrophages at the content.

[0091]

[0092] The composition for inhibiting the activation or differentiation of the above M0 macrophages into M1 macrophages and inducing differentiation into M2 macrophages can be applied as a pharmaceutical composition for preventing or treating rheumatoid arthritis.

[0093] Rheumatoid arthritis, as defined by the present invention, is a chronic autoimmune disease characterized by inflammation and proliferation of synovial cells, leading to osteoporosis and bone erosion in the bones surrounding the joints. As rheumatoid arthritis progresses, the articular cartilage gradually deteriorates, narrowing the joint space and causing a loss of tension in the joint capsule and ligaments.

[0094]

[0095] In the present invention, the term “prevention” means any act of inhibiting or delaying the onset of rheumatoid arthritis by administering the composition, and “treatment” means any act of improving or benefiting the symptoms of rheumatoid arthritis by administering the composition.

[0096]

[0097] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier.

[0098] The above "pharmaceutically acceptable carrier" may refer to a carrier or diluent that does not stimulate a living organism and does not inhibit the biological activity and properties of the compound to be injected. The type of the carrier that can be used in the present invention is not particularly limited, and any pharmaceutically acceptable carrier commonly used in the relevant technical field may be used. Non-limiting examples of the carrier include saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, etc. These may be used alone or in combination of two or more. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into an injectable formulation such as an aqueous solution, suspension, or emulsion, or into a pill, capsule, granule, or tablet.

[0099] The above pharmaceutical composition may have any one dosage form selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, oral solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories, and may be in various oral or parenteral dosage forms. When formulated, it is prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and these solid preparations are prepared by mixing one or more compounds with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerogelatin.

[0100] Additionally, the pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount.

[0101] As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dosage level can be determined according to factors including the type and severity of the individual, age, sex, type of disease, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents. And it can be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects by taking all of the above factors into consideration, and it can be easily determined by those skilled in the art.

[0102] Additionally, the pharmaceutical composition may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) depending on the intended method. For example, it may be administered parenterally.

[0103]

[0104] The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent, or may be administered together with another active ingredient exhibiting a preventive or therapeutic effect on rheumatoid arthritis. When the pharmaceutical composition of the present invention is administered together with another active ingredient, this may be referred to as "combined administration" or "combined administration." In this case, the antibody or antigen-binding fragment thereof effective in treating rheumatoid arthritis of the present invention and the other active ingredient may be administered as a single mixture or in separate forms.

[0105] In the present invention, "combined administration," "combined administration," or "combining" should be understood to refer to simultaneous, separate, sequential, or reverse administration, with no limit to the order. That is, combination is not limited to simultaneous administration, but can also be a dosage form in which the two substances act together on an individual, enabling each substance to perform its respective function at a level equal to or greater than its original function. Therefore, when the term "combined administration" is used herein, whether the administration is sequential, reverse, or separate, the order of administration is not particularly limited, and the interval between administrations of the secondary components can be such that the beneficial effects of the combination are not lost.

[0106] In the present invention, the term "subject" may be used interchangeably with subject, and refers to mammals including, but not limited to, cows, dogs, pigs, chickens, sheep, horses, and humans.

[0107]

[0108] Hereinafter, the present invention will be described in more detail through examples. These examples are intended to explain the present invention more specifically, and the scope of the present invention is not limited by these examples.

[0109]

[0110] Example 1: Confirmation of Activin A secretion from umbilical cord blood-derived mesenchymal stem cells

[0111] To profile secreted factors of umbilical cord blood-derived mesenchymal stem cells, cytokine antibody analysis (Cytokine antibody array) was performed. Specifically, after culturing MSCs in Starvation Medium, the expression of secreted proteins was profiled through Antibody Array (RayBiotech, L-1000). Here, the Antibody Array (L-1000) method is a method for detecting proteins using 1000 human antibodies. Afterwards, four inflammation and immunity-related factors, INHBA (Activin A), THBS1, GRN, and SPARC, were selected, and among them, the secretion ability of INHBA (Activin A) in umbilical cord blood-derived mesenchymal stem cells (Kangstem Co., Ltd.) was tested by 2x10 of 10 lots of umbilical cord blood-derived mesenchymal stem cells. 5 The concentration of Activin A secreted per cell was analyzed.

[0112] Specifically, umbilical cord blood-derived mesenchymal stem cells were cultured in KSB-3 basal medium (Kangstem Co., Ltd.) and seeded at 200,000 cells / ㎖ in a 24-well plate, followed by incubation in a 37℃, 5% CO2 incubator for 48 hours. At the end of incubation, 0.5 mL of the culture medium was obtained, which was centrifuged for 5 minutes to obtain the supernatant. Thereafter, the concentration of Activin A in the obtained supernatant was measured using the Activin A Quantikine ELISA kit (R&D systems).

[0113] As a result, as can be confirmed in Fig. 1, the amount of Activin A secreted from umbilical cord blood-derived mesenchymal stem cells was confirmed to be an average of 4,003.6 pg / ㎖, and the amount of Activin A secreted by lot was confirmed to be a minimum of approximately 1,844 pg / ㎖ and a maximum of 6,044 pg / ㎖ (Fig. 1).

[0114]

[0115] Example 2: Confirmation of the macrophage-regulating ability of umbilical cord blood-derived mesenchymal stem cells according to Activin A secretion

[0116] To confirm the ability of cord blood-derived mesenchymal stem cells to regulate macrophages' inflammatory factors according to their Activin A secretion ability, cord blood-derived mesenchymal stem cells (siCTL) and cord blood-derived mesenchymal stem cells with suppressed Activin A gene expression (siActivin A) were compared.

[0117] Specifically, umbilical cord blood-derived mesenchymal stem cells were seeded at 500,000 cells / ㎖ in a 100Ψ dish with KSB-3 complete Medium (Kangstem Co., Ltd.) and cultured for 24 hours in a 37℃, 5% CO2 incubator. After that, INHBA gene expression was suppressed using siRNA INHBA (Bioneer Co., Ltd.), and the amount of Activin A secretion was measured using an Activin A Quantikine ELISA kit (R&D systems Co., Ltd.).

[0118] As a result, as can be confirmed in A of Fig. 2, the secretion amount of Activin A of the cord blood-derived mesenchymal stem cells (siCTL) was measured to be approximately 6950 pg / ml, whereas the secretion amount of Activin A of the cord blood-derived mesenchymal stem cells (siActivn A) in which INHBA gene expression was suppressed was confirmed to be reduced to approximately 3795 pg / ml.

[0119] Next, to confirm the macrophage regulatory ability of cord blood-derived mesenchymal stem cells (siCTL) and cord blood-derived mesenchymal stem cells with suppressed Activin A secretion (siActivin A), co-culture with M1 macrophages or M0 macrophages was performed in a 37°C, 5% CO2 incubator for 72 hours, and the secretion of TNF-α and IL-10 was measured, respectively.

[0120] Specifically, monocytes (ATCC, THP-1 (TIB-202)) were treated with 100 ng / ml of PMA to induce differentiation into M0 macrophages, then treated with 20 ng / ml IFN-γ and 100 ng / ml LPS to induce differentiation into M1 macrophages. After culturing cord blood-derived mesenchymal stem cells for 72 hours, the amount of inflammatory factor TNF-α secretion was measured using a TNF-α Quantikine ELISA kit (R&D systems).

[0121] As a result, as can be confirmed in Figure 2B, cord blood-derived mesenchymal stem cells (siCTL) were confirmed to have TNF-α inhibitory efficacy in co-culture with M1 macrophages. In contrast, cord blood-derived mesenchymal stem cells (siActivin A) in which only Activin A secretion was inhibited did not show TNF-α inhibitory efficacy.

[0122] Through this, it was confirmed that the secretion ability of Activin A is a key factor in the regulation of pro-inflammatory factors in umbilical cord blood-derived mesenchymal stem cells in macrophages.

[0123] Additionally, after co-culture of M0 macrophages and cord blood-derived mesenchymal stem cells (siCTL or siActivin A) for 72 hours, the secretion amount of IL-10, an anti-inflammatory factor, was analyzed using the IL-10 Quantikine ELISA kit (R&D systems).

[0124] As a result, as can be confirmed in C of Fig. 2, cord blood-derived mesenchymal stem cells (siCTL) significantly promoted M2 differentiation and IL-10 secretion in co-culture with M0 macrophages, but in contrast, cord blood-derived mesenchymal stem cells siActivin A, in which only Activin A secretion was suppressed, were found to have a lower ability to promote IL-10 secretion compared to siCTL.

[0125] Through this, it was confirmed that, similar to TNF-α, Activin A secretion ability is a major factor in promoting IL-10 secretion in cord blood-derived mesenchymal stem cells (Fig. 2).

[0126]

[0127] Example 3: Confirmation of Activin A's promotion of IL-10 secretion

[0128] To confirm whether Activin A itself has the effect of promoting the secretion of anti-inflammatory factors, monocytes (ATCC, THP-1 (TIB-202)) were treated with 100 ng / ml of PMA (Sigma) to differentiate into M0 macrophages, and then treated with 20 ng / ml IL-4 and 20 ng / ml IL-13 to differentiate into M2 macrophages. Human Activin A Recombinant Protein (Peprotech) was treated at various concentrations and cultured for 72 hours in a 37°C, 5% CO2 incubator, and the amount of IL-10 secreted was confirmed using an IL-10 Quantikine ELISA kit.

[0129] As a result, as can be confirmed in Fig. 3, Activin A can be confirmed to induce differentiation of anti-inflammatory macrophages depending on the concentration and increase the secretion amount of the anti-inflammatory factor IL-10 (Fig. 3).

[0130] In particular, it was confirmed that a composition containing activin A significantly suppressed the amount of TNF-a secretion compared to a composition containing all other factors except activin A, thereby effectively suppressing the activation or differentiation into M1 macrophages, and that the amount of IL-10 secretion increased, thereby effectively promoting the differentiation of M0 macrophages into M2 macrophages (compare Figs. 2 and 3).

[0131]

[0132] Example 4: Confirmation of the immunosuppressive capacity of umbilical cord blood-derived mesenchymal stem cells secreting activin A.

[0133] To confirm the immunosuppressive capacity of cord blood-derived mesenchymal stem cells secreting Activin A, a mixed lymphocyte reaction (MLR, Mixed Lymphocyte Reaction, reference to the Mixed Lymphocyte Reaction Test Information Collection for Mesenchymal Stem Cell Therapy, October 2012, Ministry of Food and Drug Safety, National Institute of Food and Drug Safety Evaluation) experiment was conducted using peripheral blood mononuclear cells (PBMC, Goma Biotech).

[0134] Specifically, to analyze the immunosuppressive ability by confirming the proliferation and activity of lymphocytes according to the presence or absence of co-culture of mesenchymal stem cells derived from umbilical cord blood, PBMCs (1 Х 10 6 cells) were cultured on the bottom of a 24-well plate. Simultaneously, umbilical cord blood-derived mesenchymal stem cells (1.0 Х 10 5 Cells) were added to the upper insert so that the ratio of PBMC to MSC was 10:1, and the same volume of culture medium was added to the control group. After culturing for 96 hours (4 days), the proliferation of PBMC was analyzed using cell proliferation ELISA and BrdU (Roche).

[0135] As a result of this verification, as can be seen in Figure 4, the immunosuppressive capacity of umbilical cord blood-derived mesenchymal stem cells that secrete Activin A was significantly suppressed by umbilical cord blood-derived mesenchymal stem cells (Figure 4). This reconfirmed that umbilical cord blood-derived mesenchymal stem cells are associated with immunosuppressive capacity.

[0136]

[0137] Example 5: Evaluation of the efficacy of umbilical cord blood-derived mesenchymal stem cells secreting activin A in a mouse model of rheumatoid arthritis.

[0138] We evaluated the efficacy of umbilical cord blood-derived mesenchymal stem cells secreting Activin A for rheumatoid arthritis, an autoimmune disease.

[0139] Specifically, when umbilical cord blood-derived mesenchymal stem cells secreting Activin A were administered three times a week to a rheumatoid arthritis mouse model (a mouse model in which rheumatoid arthritis was induced by injecting 50 μL / mouse of 4 mg / ml Type II collagen (Chondrex) into DBA / 1 mice (Orient)), the rheumatoid arthritis index was checked (Luan J, Hu Z, Cheng J, Zhang R, Yang P, Guo H, Nan G, Guo N, Gou X. Applicability and implementation of the collagen-induced arthritis mouse model, including protocols (Review). Exp Ther Med. 2021 Sep;22(3):939.), as can be seen in Fig. 5 A and B, it was confirmed that the efficacy of significantly suppressing rheumatoid arthritis was confirmed compared to the control group, and in particular, it was confirmed that the efficacy was the best when administered together with MTX, a representative drug for the treatment of rheumatoid arthritis.

[0140] In addition, analysis of foot tissues of a mouse model of rheumatoid arthritis using Hematoxylin and Eosin (H&E) and Safranin O staining confirmed the effect of suppressing inflammatory cell infiltration, bone erosion, and cartilage damage (Fig. 5C).

[0141]

[0142] Example 6: Evaluation of the efficacy of Activin A in a mouse model of rheumatoid arthritis

[0143] Next, the efficacy of Activin A on rheumatoid arthritis was evaluated. Human Activin A Recombinant Protein was administered three times a week to a rheumatoid arthritis mouse model, and the arthritis index was checked. As shown in Fig. 6, it was confirmed that there was a significant inhibitory effect compared to the control group when Activin A was treated. In addition, the results of analyzing the foot tissue of a rheumatoid arthritis mouse model through Hematoxylin and Eosin (H&E) staining confirmed that Activin A had the effect of inhibiting inflammatory cell infiltration, bone erosion, and cartilage damage (Fig. 6).

[0144]

[0145] Example 7: Confirmation of the critical point of Activin A through its TNF-a secretion inhibition effect.

[0146] To determine the concentration at which Activin A has an immunosuppressive effect in rheumatoid arthritis, the critical point of Activin A was identified through the TNF-a secretion inhibition effect.

[0147] Macrophages are well known to play a central role in the pathophysiology of rheumatoid arthritis (RA), secreting inflammatory cytokines and tissue-degrading enzymes that induce inflammation and tissue destruction. Among these, M1 macrophages are generally activated by interferon-γ (IFN-γ) or lipopolysaccharide (LPS), producing proinflammatory cytokines (e.g., tumor necrosis factor-α, interleukin-1b, and interleukin-6), and initiating immune responses by eliminating microorganisms through phagocytosis.

[0148] In consideration of this, in order to reproduce the above phenomenon in vitro, monocytes (ATCC, THP-1 (TIB-202)) were differentiated from M0 macrophages by treating with 100 ng / ml of PMA and then with 20 ng / ml of IFN-γ and 100 ng / ml of LPS, and in this environment, Activin A-secreting umbilical cord blood-derived mesenchymal stem cells were co-cultured or a composition containing Activin A secreted from umbilical cord blood-derived mesenchymal stem cells was treated at various concentrations from 0.1 pg / ml to 20,000 pg / ml to confirm the degree of inhibition of TNF-a, a representative factor secreted from M1 macrophages. The concentration was confirmed as a 10-fold increase in the range of 1,000 pg / ml or less, and a 2-fold increase in the range of 10,000 pg / ml and 20,000 pg / ml (see Fig. 7).

[0149] As a result, as can be confirmed in Fig. 8, it was confirmed that the composition containing Activin A suppressed the inflammatory factor TNF-a at a concentration of 0.1 pg / ml or more and less than 20,000 pg / ml under M1 macrophage differentiation induction conditions, and among them, the effective TNF-a inhibition ability was confirmed in the range of 10,000 pg / ml or less. In contrast, it was confirmed that the secretion of TNF-a was significantly increased and cytotoxicity was present at a concentration of 20,000 pg / ml or more, and in particular, it was confirmed that the increase was significantly greater than when compared to PC (M1), and it was confirmed that the effective concentration range for inhibiting M1 macrophage differentiation of the composition containing Activin A of the present invention was 0.1 pg / ml or more and less than 20,000 pg / ml, and among them, 0.1 pg / ml or more and 10,000 pg / ml or less was an effective concentration range.

[0150]

[0151] Example 8: Efficacy test of umbilical cord blood-derived mesenchymal stem cells secreting Activin A or a composition containing Activin A_Efficacy of promoting IL-10 secretion

[0152] M2 macrophages are differentiated and activated by specific cytokines, such as IL-4 and IL-13. M2 macrophages produce anti-inflammatory cytokines and induce tissue remodeling, promoting wound healing and tissue repair.

[0153] In consideration of this, monocytes (ATCC, THP-1 (TIB-202)) were co-cultured with cord blood-derived mesenchymal stem cells that secrete Activin A in the M0 macrophage state through PMA treatment of 100 ng / ml in vitro, or compositions containing Activin A secreted from cord blood-derived mesenchymal stem cells were treated at various concentrations from 0.1 pg / ml to 20,000 pg / ml to confirm the secretion level of IL-10, a representative factor secreted from M2 macrophages. The secretion amount of IL-10, a representative factor secreted during M2 differentiation, was measured using the previously used kit to confirm the efficacy of inducing differentiation into M2 macrophages.

[0154] As a result, as can be confirmed in Fig. 9 and Fig. 3, the effectiveness of IL-10 secretion by co-culture of M0 macrophages and umbilical cord blood-derived mesenchymal stem cells was confirmed to be in the range of 1:0.01 to 1:2 in terms of co-culture ratio, and it was confirmed that there was no effectiveness in IL-10 secretion outside that range.

[0155] In addition, when treating M0 macrophages with a composition (CM) containing Activin A, the effectiveness of IL-10 secretion was confirmed, and it was confirmed to be significant in the range of 0.1 to 10,000 pg / ml, and especially in the range of 0.1 to 5,000 pg / ml. When treating M0 macrophages with a composition containing Activin A at a concentration of 20,000 pg / ml, which is outside of 5,000 pg / ml, it was confirmed that there was no significance in IL-10 secretion.

[0156] From the perspective of cord blood-derived mesenchymal stem cells that secrete Activin A, if the amount of Activin A secreted is converted into the number of cells based on the test method and measurement results in Figure 1C, and calculated by converting it into the average number of cells of cord blood-derived mesenchymal stem cells (converting the amount secreted under general conditions without stimulation, not under specific conditions), it is 1:0.01 (1 x 10) that Activin A is secreted at 5 pg / ml or more per 1,000 cells. 4 )~1:2 (2 x 10 6 ) the secretion amount of Activin A can be calculated as a minimum of 50 to 10,000 pg / ml, and in the average measurement range, it can be calculated as 200 to 40,000 pg / ml.

[0157]

[0158] In contrast, unlike cord blood-derived mesenchymal stem cells that secrete Activin A in M0 macrophages or compositions containing Activin A, Activin A alone was found to have no effect on IL-10 secretion in M0 macrophages.

[0159] However, under conditions inducing M2 macrophage differentiation, it was confirmed that Activin A alone significantly increased IL-10 secretion in a concentration-dependent manner.

[0160] M2 macrophage differentiation was induced by adding IL-4 and IL-13 cytokines to M0 macrophages. Treatment of Activin A in an environment inducing M2 macrophage differentiation can be seen as treating IL-4, IL-13, and Activin A during M0 macrophage culture. In other words, the above results can be interpreted as indicating that Activin A alone does not have the effect of inducing M2 differentiation, and that it has the effect of boosting the differentiation of M2 macrophages in an M2 macrophage differentiation environment.

[0161] Through this, M0 macrophages are undifferentiated cells that differentiate into M1 or M2 depending on the environment and are activated according to their characteristics. Rheumatoid arthritis is a disease in which a continuous immune activation response occurs, M1 macrophage differentiation, and excessive secretion of pro-inflammatory cytokines occur, causing chronic inflammation, cartilage damage and tissue destruction, and pain responses. It can be said that the differentiation conditions environment of M1 macrophages is already similar to the onset of rheumatoid arthritis.

[0162] In this respect, it can be seen that a composition containing activin A or cord blood-derived mesenchymal stem cells secreting the composition contain various cytokines, thereby inhibiting the activity or differentiation of M1 macrophages in a rheumatoid arthritis state, thereby reducing the secretion of the inflammatory cytokine TNF-a, and at the same time effectively inducing the differentiation of M0 macrophages into M2 macrophages, thereby effectively treating rheumatoid arthritis due to the immune modulation effect and homeostasis maintenance.

[0163]

[0164] In summary, a composition containing activin A or umbilical cord blood-derived mesenchymal stem cells secreting it can be developed into a pharmaceutical agent for cell therapy for rheumatoid arthritis.

[0165]

[0166] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. A composition comprising activin A, or a composition comprising mesenchymal stem cells secreting the composition as an active ingredient, for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation into M2 macrophages.

2. A composition for suppressing the activity or differentiation of M1 macrophages and inducing differentiation into macrophages, characterized in that the composition containing activin A or the cord blood-derived mesenchymal stem cells secreting the composition in the first paragraph suppresses the differentiation of M1 macrophages while simultaneously inducing the differentiation of M0 macrophages into M2 macrophages, thereby regulating immunity.

3. In the first paragraph, the composition comprising activin A comprises activin A, THBS1, GRN, and SPARC, a composition for inhibiting the activation or differentiation of M1 macrophages and inducing the differentiation of M0 macrophages into M2 macrophages.

4. A composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation of M0 macrophages into M2 macrophages, wherein the composition comprising activin A in the first paragraph further comprises at least one selected from the group consisting of VEGF, EDA, MIF, TIMP-2, and TGF-b1.

5. A composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation of M0 macrophages into M2 macrophages, wherein the composition containing activin A or the mesenchymal stem cells secreting the composition in the first paragraph are co-cultured with M0 macrophages at a ratio of 0.01 to 2:

1.

6. A composition for inhibiting the activation or differentiation of M1 macrophages and inducing the differentiation of M0 macrophages into M2 macrophages, wherein the composition containing activin A in the first paragraph is 0.1 pg / ml or more and less than 20,000 pg / ml.

7. A composition for inhibiting the activation or differentiation of M1 macrophages and inducing the differentiation of M0 macrophages into M2 macrophages, wherein the composition containing activin A in the first paragraph is 1,000 pg / ml or more and less than 20,000 pg / ml.

8. A composition for inhibiting the activation or differentiation of M1 macrophages and inducing the differentiation of M0 macrophages into M2 macrophages, wherein the composition containing activin A in the first paragraph is 1,000 pg / ml or more and less than 10,000 pg / ml.

9. A composition for inhibiting the activation or differentiation of M1 macrophages and inducing the differentiation of M0 macrophages into M2 macrophages, wherein the composition containing activin A in the first paragraph is 1,000 pg / ml or more and less than 5,000 pg / ml.

10. A composition for inhibiting the activation or differentiation of M1 macrophages and promoting the differentiation of M0 macrophages into M2 macrophages, characterized in that the activin A in the first paragraph is an M0 macrophage or boosts the differentiation of M2 macrophages in an M2 macrophage differentiation environment.

11. A composition for inhibiting the activity or differentiation of M1 macrophages and promoting the differentiation of M0 macrophages into M2 macrophages, wherein the M2 macrophage differentiation environment in paragraph 10 is an environment containing IL-4 and IL-13, or an environment in which rheumatoid arthritis has developed.

12. A composition for inhibiting the activity or differentiation of M1 macrophages and inducing differentiation of M0 macrophages into M2 macrophages, wherein the mesenchymal stem cells in paragraph 1 are mesenchymal stem cells derived from umbilical cord blood.

13. A method for inhibiting the activation or differentiation of M1 macrophages and inducing differentiation of M0 macrophages into M2 macrophages, comprising a step of treating a composition containing activin A or mesenchymal stem cells secreting the composition.

14. A pharmaceutical composition for preventing or treating rheumatoid arthritis, comprising a composition according to any one of claims 1 to 12.

15. A pharmaceutical composition according to claim 14, wherein the composition further comprises a pharmaceutically acceptable carrier.

16. A pharmaceutical composition according to claim 15, wherein the pharmaceutically acceptable carrier further comprises at least one selected from the group consisting of a preservative, a solubilizer, a stabilizer, an excipient, a lubricant, a preservative, and an additive.

17. A pharmaceutical composition according to claim 14, characterized in that the composition containing activin A or the cord blood-derived mesenchymal stem cells secreting the composition suppresses the activation and differentiation of M1 macrophages and simultaneously induces the differentiation of M0 macrophages into M2 macrophages, thereby regulating immunity.

18. A method for preventing or treating rheumatoid arthritis, comprising administering to a subject the pharmaceutical composition of Article 14.

19. A composition for boosting the induction and differentiation of M0 macrophages into M2 macrophages in an M2 macrophage differentiation environment, comprising activin A as an active ingredient.

20. A composition for boosting M2 macrophage differentiation in claim 19, wherein the M2 macrophage differentiation environment is an environment containing IL-4 and IL-13, or an environment in which rheumatoid arthritis has developed.

Images