Composition for treating osteoarthritis

Abstract

The present invention relates to a composition for disease-modifying treatment of osteoarthritis. More specifically, the present invention relates to a composition for treating osteoarthritis and a method for treating osteoarthritis by using same, the composition comprising human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) and an acellular cartilage matrix.

Description

Composition for treating osteoarthritis

[0001] The present invention relates to a composition for treating osteoarthritis. More specifically, it relates to a composition for treating osteoarthritis comprising human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) and an acellular cartilage matrix, and a method for treating osteoarthritis using the same.

[0002]

[0003] Osteoarthritis is a representative degenerative joint disease caused by various risk factors such as increasing age, obesity, trauma, and overuse. Among these, knee osteoarthritis is characterized by progressive damage to articular cartilage, changes in the subchondral bone, synovitis, and osteophyte formation. Osteoarthritis is known as a major disease that not only significantly reduces patients' quality of life by causing chronic pain, limited joint movement, and functional decline, but also rapidly increases medical costs and socioeconomic burdens in an aging society.

[0004] Currently, primary treatments for osteoarthritis consist of non-steroidal anti-inflammatory drugs (NSAIDs), symptom relievers such as acetaminophen, intra-articular hyaluronic acid or steroid injections, physical therapy, weight loss, and exercise therapy. However, these conservative treatments are mainly limited to pain relief and temporary functional improvement, and have limitations in fundamentally regenerating already damaged cartilage tissue or inhibiting the progression of the disease in the long term. In patients with moderate to severe (Kellgren-Lawrence (K&L) grade 2-4) conditions, there are many cases where total knee arthroplasty is eventually required, and this also presents issues such as surgical invasiveness, the lifespan of the artificial joint, and the need for reoperation.

[0005] To address these limitations, surgical and cellular therapeutic approaches have been developed, such as autologous chondrocyte implantation, cell therapy using autologous bone marrow-derived or adipose-derived stem cells, and microfracture. Allogeneic cord blood-derived mesenchymal stem cell therapies for cartilage damage (e.g., allogeneic cord blood-derived stem cell therapies for knee cartilage defects) have already secured clinical application and patent rights, and these therapies work by inducing cartilage regeneration through the transplantation of cells into the cartilage defect site. However, these therapies generally require surgical procedures to expose the cartilage defect lesion and attach and fix the cells, and they have limitations in terms of invasiveness and cost for convenient application to the majority of patients with extensive arthritis lesions or K&L grades 2-3.

[0006]

[0007] Meanwhile, intra-articular cell injection therapies, involving the direct injection of mesenchymal stem cells into the joint cavity, have also been reported in animal studies and some clinical trials. While approaches involving the intra-articular injection of cord blood, bone marrow, or adipose-derived MSCs alone have shown improvements in pain and functional indicators (KOOS, WOMAC, VAS, etc.) in some studies, they have limitations in that they have failed to demonstrate consistent, significant improvements in structural indicators (K&L grade, MRI-based cartilage thickness, MOCART, WORMS scores, etc.) during long-term follow-up. Furthermore, it has been pointed out that injecting only cells makes it difficult to prove a true disease-modifying osteoarthritis drug (DMOAD) effect, as sufficient cell engraftment and long-term survival are not ensured in terms of the intra-articular mechanical environment and interaction with the subchondral bone.

[0008] Recently, the concept of convergence advanced biopharmaceuticals combining stem cells and cartilage matrix has been proposed, and it is reported that osteoarthritis treatments utilizing umbilical cord blood-derived mesenchymal stem cells and acellular cartilage matrix together are under development. Non-clinical data suggests that these technologies can increase the cartilage regeneration effect compared to cell administration alone by providing a three-dimensional scaffold that supports the differentiation and cartilage regeneration of stem cells. However, conventional technologies are based mainly on results from animal experiments or on human clinical data that quantitatively and qualitatively integratedly evaluated structural improvements (e.g., improvement in MOCART score, reduction in WORMS score, improvement in K&L grade, etc.) in clinical trials. Consequently, they do not specify the disease-modifying effect of simultaneously improving pain, function, and structure with a single intra-articular administration, linked to a specific dosage range and patient group (such as patients with knee osteoarthritis of K&L grade 2-3).

[0009]

[0010] Therefore, for patients with osteoarthritis, there is still an urgent need to develop a treatment linked to specific dosage ranges and patient groups that provides disease-modifying therapeutic effects, simultaneously improving pain, function, and structure through intra-articular administration alone.

[0011]

[0012] One objective of the present invention is to provide a pharmaceutical composition for treating osteoarthritis comprising human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) and cartilage acellular matrix (CAM).

[0013] Another objective of the present invention is to provide a formulation for intra-articular administration for the treatment of osteoarthritis comprising human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) and cartilage acellular matrix (CAM).

[0014] Another objective of the present invention is to provide a method for treating osteoarthritis comprising the step of administering the pharmaceutical composition or administration preparation to an individual.

[0015] Another objective of the present invention is to provide a method for promoting cartilage or subchondral bone remodeling, comprising the step of administering the pharmaceutical composition or administration preparation to an individual.

[0016] Another objective of the present invention is to provide a kit for preparing an intra-articular administration agent for the treatment of knee osteoarthritis, comprising: a first vial containing umbilical cord blood-derived mesenchymal stem cells; and a second vial containing a chondrocellular matrix.

[0017] Another objective of the present invention is to provide a method for preparing a pharmaceutical composition for treating osteoarthritis comprising human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) and cartilage acellular matrix (CAM).

[0018] Another objective of the present invention is to provide a pharmaceutical composition comprising human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) and cartilage acellular matrix (CAM) for the treatment of osteoarthritis.

[0019] Another objective of the present invention is to provide a kit comprising human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) and cartilage acellular matrix (CAM) for the treatment of osteoarthritis.

[0020] Another objective of the present invention is to provide a use for manufacturing an intra-articular administration agent for the treatment of knee osteoarthritis using a kit comprising: a first vial containing umbilical cord blood-derived mesenchymal stem cells; and a second vial containing a chondrocellular matrix.

[0021]

[0022] The disease-modifying therapeutic composition for osteoarthritis according to the present invention exhibits a disease-modifying effect by inducing structural recovery at the level of cartilage and subchondral bone, along with improvement of pain and function, when administered as a single dose into the joint cavity of an osteoarthritis patient. In particular, for knee osteoarthritis patients with cartilage defects of Kellgren-Lawrence (K&L) grade 2 or 3 and ICRS grade 3 or 4 on MRI, a specific range of doses, namely 5.0 × 10⁻⁶ 7 Up to 1.0×10 8 When a composition comprising hUCB-MSC and 40 to 60 mg of CAM is administered as a single intra-articular dose, clinical indicators such as VAS, WOMAC, IKDC, and KOOS are significantly improved at 24 weeks after administration, and in MRI-based MOCART and WORMS evaluation indicators, the cartilage or subchondral bone structure is stabilized and partially remodeled, such as a reduction in subchondral bone marrow edema and normalization of the subchondral bone boundary, and furthermore, the pattern of changes in biomarkers, such as a decrease in IL-1β and TNF-α, an increase in PIIANP, and the stabilization of CTX-II and COMP / MMP-3, promotes the reduction of intra-articular inflammation and cartilage synthesis, so the composition of the present invention can be usefully utilized as a disease-modifying therapeutic agent capable of inhibiting or reversing the progression of osteoarthritis with only a single intra-articular dose.

[0023]

[0024] FIG. 1 is a schematic diagram illustrating the mechanism by which cord blood-derived mesenchymal stem cells and chondrocellular matrix (OSCA), which are the disease-modifying therapeutic composition for osteoarthritis according to the present invention, act within an osteoarthritis joint to induce cartilage regeneration, subchondral bone stabilization, and inflammation control.

[0025] FIG. 2 is a schematic diagram illustrating the overall design (test group composition, administration time, and follow-up schedule) of the K0701 Phase 1 clinical trial conducted using the composition of the present invention.

[0026] FIG. 3 is a diagram showing the trial protocol (see COLINICALTRIUAL.GOV) and visit flow, including administration, safety evaluation, clinical indicator evaluation, and imaging examination at each visit time in the Phase 1 clinical trial of K0701 using the composition of the present invention.

[0027] Figure 4 is a graph showing the changes in pain (VAS) and functional indicators (WOMAC, IKDC, KOOS) by dose cohort up to 24 weeks after a single intra-articular administration of the composition of the present invention to a patient with knee osteoarthritis.

[0028] Figure 5 is a diagram showing the changes in MOCART scores and WORMS scores by cohort in patients with knee osteoarthritis between baseline and 24 weeks after a single intra-articular administration of the composition of the present invention.

[0029] Figure 6 is a figure showing a comparison of structural changes in the subchondral bone, such as filling of cartilage defects and formation of regenerative tissue, reduction of subchondral bone marrow edema, and normalization of the subchondral bone boundary, by taking MRIs of the knee joint before administering the composition of the present invention once into the joint cavity and at 24 weeks after administration.

[0030] Figure 7 is a figure showing the changes in the concentrations of biomarkers such as IL-1β, TNF-α, PIIANP, CTX-II, COMP, and MMP-3 measured in patients with knee osteoarthritis for 24 weeks after a single intra-articular administration of the composition of the present invention, by time course and dose cohort.

[0031]

[0032] The specific details for implementing the present invention are described as follows. Meanwhile, each description and embodiment disclosed herein may be applied to other descriptions and embodiments. That is, all combinations of the various elements disclosed herein fall within the scope of the present invention. Furthermore, the scope of the present invention is not to be limited by the specific descriptions provided below.

[0033] Meanwhile, unless the context otherwise requires in this specification, expressions such as “include,” “comprising,” “containing,” etc., should be understood to mean the inclusion of a specified integer or group of integers, but not to exclude other integers or sets of integers.

[0034]

[0035] Current drug and injection therapies for osteoarthritis, particularly knee osteoarthritis, are limited to symptom relief such as pain and functional improvement, and fail to sufficiently achieve fundamental recovery of already damaged articular cartilage and subchondral bone structures or inhibition of disease progression. Additionally, while the method of injecting mesenchymal stem cells of various origins, including human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC), into the joint cavity alone has reported improvements in pain and functional indicators in some clinical studies, it has limitations in that it has not demonstrated consistent significant improvement in structural indicators (e.g., MRI-based cartilage thickness, cartilage defect volume, subchondral bone changes, etc.) during long-term follow-up, and the dosage, administration method, and target patient group have not been sufficiently established.

[0036] In addition, surgical treatments involving the transplantation of allogeneic umbilical cord blood-derived mesenchymal stem cells and hyaluronic acid hydrogel or other scaffolds into cartilage defects have been reported to demonstrate cartilage regeneration and long-term safety in some knee osteoarthritis patients with localized cartilage defects. However, these methods are highly invasive as they require open or arthroscopic surgery, making them difficult to apply broadly to patients with general degenerative arthritis with extensive lesions. Furthermore, since these techniques are primarily focused on transplantation therapy for localized cartilage defects, they have not been able to clearly demonstrate the disease-modifying osteoarthritis drug (DMOAD) effect of simultaneously inducing pain and functional improvement as well as structural improvement of the joint (remodeling of cartilage and subchondral bone) through a single intra-articular administration in typical osteoarthritis patients with Kellgren-Lawrence (K&L) grade 2 to 3.

[0037] Furthermore, although hybrid formulations combining cartilage acellular matrix (CAM) and mesenchymal stem cells have recently been reported in animal models and early clinical studies, these studies also lack sufficient established technology specifically defined as a use and treatment method for knee osteoarthritis by integrating clinical characteristics such as K&L grade, age, and body mass index with MRI-based quantitative indicators (MOCART, WORMS, etc.), and (iv) the appropriate dosage range verified in clinical trials, (ii) the maximum effect achievable solely through intra-articular administration, (iii) data identifying structural improvement effects by integrating clinical characteristics such as K&L grade, age, and body mass index with MRI-based quantitative indicators (MOCART, WORMS, etc.).

[0038] In this regard, the present invention is based on the novel identification of a pharmaceutical composition comprising a composite preparation containing umbilical cord blood-derived mesenchymal stem cells and an acellular cartilage matrix, which not only exhibits excellent safety and tolerability when administered as a single intra-articular injection to patients with knee osteoarthritis, but also shows significant improvement in pain and functional indicators (KOOS, WOMAC, VAS, IKDC, etc.) as well as improvement in cartilage and subchondral bone structure through MRI and radiological indicators (MOCART, WORMS, K&L grade, etc.).

[0039]

[0040] One aspect of the present invention for solving the above problem provides a pharmaceutical composition for treating osteoarthritis comprising umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) and cartilage acellular matrix (CAM).

[0041] In one embodiment, the hUCB-MSC is 2.5 x 10 per single administration 7 Up to 1.0X10 8 It may be included in the number of cells.

[0042] In one embodiment, the hUCB-MSC is 5.0X10 7 Up to 1.0X10 8 It may be included in the number of cells.

[0043] In one embodiment, the CAM may be included in an amount of 40 to 80 mg per single administration.

[0044] In one embodiment, the CAM may be included in an amount of 50 to 70 mg per single administration.

[0045] In one embodiment, the osteoarthritis may be knee osteoarthritis.

[0046] In one embodiment, the composition may be formulated in the form of a suspension having a total dosage of 2.0 mL to 4.5 mL.

[0047] In one embodiment, the composition comprises: i) 2.5X10 7 ii) a 2.25 mL formulation containing hUCB-MSC and 60 mg of CAM, 5.0X10 7 A 3.0 mL formulation containing hUCB-MSC and 60 mg of CAM, or iii) 1.0×10 8 It may be a 4.5 mL formulation containing hUCB-MSC and 60 mg of CAM.

[0048] In one embodiment, the composition may further comprise one or more selected from the group consisting of a cryopreservation agent, an isotonic solution, and a buffer solution.

[0049] In one embodiment, the composition may be used by mixing the hUCB-MSC preparation and the CAM preparation in a single syringe immediately before administration.

[0050] In one embodiment, the composition may be used for injection into the joint cavity.

[0051] In one embodiment, the composition may be characterized by being designed to be injected into the joint cavity through a supererolateral approach in the affected knee joint of a patient with knee osteoarthritis.

[0052] In one embodiment, the knee osteoarthritis patient may be a knee osteoarthritis patient with Kellgren-Lawrence grade 2 or 3 on X-ray examination, or a knee osteoarthritis patient with localized cartilage defects of grade 3 or 4 according to International Cartilage Repair Society (ICRS) standards on magnetic resonance imaging (MRI).

[0053] In one embodiment, the knee osteoarthritis patient may be a patient who complains of pain of 50 mm or more on a 100 mm VAS scale despite conservative treatment including non-steroidal anti-inflammatory drugs, physical therapy, or exercise therapy for at least 12 weeks.

[0054] In one embodiment, the composition may result in a decrease in the VAS pain score by 50% or more, an increase in the MOCART score by 70%, a decrease in the WORMS cartilage subscore by 40%, an increase in the IKDC score by 50% or more, a decrease in the WOMAC total score by 50% or more, or an increase in the KOOS Quality of Life (QoL) subscale by 50% or more compared to baseline at 24 weeks after administration.

[0055] In one embodiment, at 24 weeks after administration of the composition, the proportion of responders meeting the OMERACT-OARSI criteria of a relative improvement rate of 50% or more and an absolute change of 20 points or more may be 50% or more, or the proportion of responders in one or more scales based on KOOS may be 30% or more, or the proportion of responders based on VAS may be 60% or more.

[0056] In one embodiment, the composition may be characterized in that, at 24 weeks after administration, the subchondral bone structure does not deteriorate compared to the baseline or the subchondral bone structure improves.

[0057] In one embodiment, the improvement of the subchondral bone structure may include normalization or reduction of bone marrow signals, restoration of continuity of the subchondral bone boundary, or cartilage or subchondral bone remodeling.

[0058] In one embodiment, at 24 weeks after administration of the composition, compared to baseline, it may have one or more features selected from the group consisting of (a) a decrease in serum IL-1β concentration, (b) a decrease in serum TNF-α concentration, (c) an increase in serum PIIANP concentration, (d) maintenance of urine CTX-II (creatinine corrected) concentration, and (e) maintenance of serum COMP and MMP-3 concentrations.

[0059]

[0060] The present invention will be described in detail below.

[0061]

[0062] The term "umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC)" used in the present invention refers to adult mesenchymal stem cells isolated and cultured from human umbilical cord blood, and typically includes cells that are positive for surface markers such as CD73, CD90, and CD105, and negative for hematopoietic lineage markers such as CD34, CD45, and HLA-DR. The mesenchymal stem cells have the ability to differentiate into three lineages, such as osteocytes, chondrocytes, and adipocytes, and can contribute to cartilage protection and regeneration, inflammation control, and pain relief by secreting immunomodulatory factors and various growth factors / cytokines.

[0063] In the pharmaceutical composition of the present invention, the hUCB-MSC is 2.5 x 10 per single administration 7 Up to 1.0X10 8 It can be included as a number of cells. Specifically, 5.0 x 10 7 Up to 1.0X10 8 It can be included as a number of cells, and more specifically, 2.5X10 7 , 5.0Х10 7 , 1.0Х10 8 It can be included in the number of cells.

[0064] The number of cells and the amount of chondrocyte-free matrix included in the above hUCB-MSC per single administration may be increased or decreased within a general range depending on the patient's condition, and may be selected and determined by a conventional clinician or a skilled expert to be within a safe and effective range.

[0065] The above hUCB-MSC is not particularly limited but can be manufactured under clinical-grade GMP conditions and can be used after thawing following cryopreservation.

[0066]

[0067] The term "chondrocellular matrix (CAM)" as used in the present invention refers to an extracellular matrix obtained by decellularizing cartilage tissue derived from animals or humans to remove cells and nucleic acids. Specifically, the chondrocellular matrix may comprise a solid or powder-form matrix obtained by removing cellular components from allogeneic or xenogeneic cartilage tissue and primarily containing cartilage matrix components such as collagen, proteoglycans, and glycosaminoglycans. The CAM may be provided in the form of freeze-dried powder or gel and can serve as a support structure to which cells can attach and maintain survival and differentiation when mixed with hUCB-MSC.

[0068] The above cartilage acellular matrix may be used in combination with cartilage acellular matrix (CAM), which may be prepared by isolating cartilage from animals and decellularizing it through enzymatic or physicochemical methods as in known processes, or may use commercially available materials, but is not limited thereto.

[0069] The above CAM may be in various forms, such as powder or sponge, and is not particularly limited thereto.

[0070] In addition, the animal capable of obtaining the above CAM is not particularly limited as long as it is an animal having cartilage tissue, but specifically, it may be a pig.

[0071] In the pharmaceutical composition of the present invention, the CAM may be included in an amount of 40 to 80 mg per single administration. Specifically, it may be included in an amount of 50 to 70 mg per single administration, and more specifically, it may be 55 to 65 mg, 56 to 65 mg, 57 to 65 mg, 59 to 65 mg, 55 to 64 mg, 55 to 63 mg, 55 to 62 mg, 55 to 61 mg, and even more specifically, 60 mg.

[0072] For example, the above-mentioned cartilage acellular matrix may be provided as an injectable formulation suspended in an aqueous carrier such as physiological saline, and may be processed so as to minimize immune responses caused by cell residues during the acellularization process.

[0073] For example, an injectable formulation containing the above-mentioned cartilage acellular matrix may be manufactured according to Korean Patent Publication No. 10-2021-0165676.

[0074]

[0075] The term "osteoarthritis" as used in this invention refers to a representative chronic degenerative joint disease characterized by the gradual wear and tear of articular cartilage, changes in subchondral bone, the formation of osteophytes, and synovial inflammation. It is primarily caused by a combination of risk factors such as aging, overweight, joint trauma, mechanical overload, and misalignment, and results in pain, reduced range of motion, functional impairment, and a decline in quality of life. Existing drugs, injections, and physical therapy focus mainly on relieving pain and inflammation, and thus have limitations in fundamentally inhibiting or reversing the progression of joint structure.

[0076] Among these, "knee osteoarthritis" refers to a degenerative disease in which osteoarthritis occurs in the knee. The above knee osteoarthritis may include primary and secondary types classified based on cause, and medial compartment type, lateral compartment type, patellofemoral type, and three-compartment type classified based on location. Specifically, depending on the degree of progression, it may refer, for example, to knee osteoarthritis classified as K&L grade 2 or 3 among degenerative joint diseases classified as Kellgren-Lawrence (K&L) grade 1 to 4 on radiographic examination; for another example, it may refer to osteoarthritis with localized cartilage defects of International Cartilage Repair Society (ICRS) grade 3 or 4 on MRI; or, for yet another example, it may include knee osteoarthritis that complains of pain ≥50 mm on the VAS 100 mm scale and does not show sufficient improvement even with conservative treatment for more than 12 weeks.

[0077] Another example is knee osteoarthritis classified as K&L grade 2 or 3 among degenerative joint diseases classified as Kellgren-Lawrence (K&L) grade 1 to 4 on radiographic examination, and knee osteoarthritis with localized cartilage defects of International Cartilage Repair Society (ICRS) grade 3 or 4 on MRI.

[0078] Another example may be knee osteoarthritis classified as K&L grade 2 or 3 among degenerative joint diseases classified as Kellgren-Lawrence (K&L) grade 1 to 4 on radiographic examination, knee osteoarthritis with localized cartilage defects of grade 3 or 4 according to International Cartilage Repair Society (ICRS) standards on MRI, and knee osteoarthritis that complains of pain ≥50 mm on the VAS 100 mm scale and does not show sufficient improvement even with conservative treatment for more than 12 weeks.

[0079] As used in the present invention, the term "drug for disease modification" refers to a drug that does not merely alleviate pain or improve function, but fundamentally alters the rate of disease progression or the mechanism of the disease to slow down or inhibit structural deterioration. Specifically, a drug for disease modification in osteoarthritis may include a drug that accompanies changes in the sub-joint structure, such as cartilage or subchondral bone remodeling manifested by an increase in MRI-based MOCART scores and / or a decrease in WORMS scores such as cartilage subscores and / or the stabilization or improvement of K&L grades, as well as preservation (delay in deterioration) and recovery.

[0080] The above "MOCART score" is the Magnetic Resonance Observation of Cartilage Repair Tissue 2.0 score, which is an MRI index that evaluates items such as cartilage defect filling, regenerated tissue integration, surface, structure, signal intensity, and subchondral bone changes, and the higher the score, the closer the cartilage condition is to normal.

[0081] The aforementioned "WORMS score" stands for Whole-Organ Magnetic Resonance Imaging Score, which refers to a semi-quantitative MRI indicator evaluating the entire structure of the knee joint, including cartilage, subchondral bone marrow lesions, osteophytes, synovitis, menisci, ligaments, and intra-articular loose bodies; a lower score indicates a structural condition closer to normal.

[0082] In each of the above “VAS, IKDC, WOMAC, KOOS” cases, they are tools for evaluating pain and function in patients; VAS (Visual Analog Scale) measures the degree of pain by marking it on a line from 0 to 100 mm, IKDC (International Knee Documentation Committee) evaluates the patient’s subjective symptoms, objective function, and sports activities, WOMAC (Western Ontario and McMaster Universities Osteoarthritis Index) is an evaluation index consisting of pain, stiffness, and function items, and KOOS (Knee injury and Osteoarthritis Outcome Score) is an OA evaluation index including five subscales of pain, symptoms, daily life, sports and leisure, and quality of life.

[0083]

[0084] The above composition may be formulated in the form of a suspension with a total dosage of 2.0 mL to 4.5 mL per single dose.

[0085] The term "administrative dose" above refers to the amount of a pharmaceutical composition injected into the knee joint cavity in a single dose; specifically, it may be formulated in the form of a suspension ranging from 2.0 mL to 4.5 mL per single administration. If the pharmaceutical composition of the present invention is less than 2.0 mL, it may be difficult to deliver a sufficient amount of cells and chondrocyte-free matrix into the joint cavity, and if it exceeds 4.5 mL, the risk of side effects such as pain, discomfort, or increased exudate due to increased intra-articular pressure may increase. Therefore, the above volume range is based on the number of cells (e.g., 5.0 x 10⁻¹⁰). 7 Up to 1.0X10 8 It may be a pharmaceutically reasonable range to stably suspend the cell-free matrix of cartilage (e.g., 60 mg) while minimizing physical burden when injected into the joint cavity.

[0086] In addition, the above suspension can be mixed and suspended immediately before administration and administered as a single intra-articular injection, which may be advantageous in terms of preparation, handling, and patient compliance in clinical settings.

[0087] As an example, the pharmaceutical composition of the present invention comprises: i) 2.5X10 7 ii) a 2.25 mL formulation containing hUCB-MSC and 60 mg of CAM, 5.0X10 7 A 3.0 mL formulation containing hUCB-MSC and 60 mg of CAM, or iii) 1.0×10 8 It may be a 4.5 mL formulation containing hUCB-MSC and 60 mg of CAM.

[0088]

[0089] The pharmaceutical composition of the present invention can be used by mixing the hUCB-MSC preparation and the CAM preparation in a single syringe immediately before administration.

[0090] The pharmaceutical composition of the present invention can be used by storing a preparation containing umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) and a preparation containing chondrocyte-free matrix (CAM) in separate containers immediately before administration, and then mixing them in a single syringe at the time of administration. This method of mixing immediately before administration has the advantage of preventing a decrease in stability during long-term storage due to physical and chemical interactions between the cells and CAM, and allowing the preparation to be administered to the patient while maintaining the cell viability and structural characteristics of the CAM in an optimal state. Furthermore, by injecting the mixture directly into the joint cavity after mixing in a single syringe, the simplification of the preparation process, the reduction of the risk of contamination, and the convenience of handling by the clinician can be simultaneously ensured.

[0091] The pharmaceutical composition of the present invention may be for intra-articular injection. Here, "intra-articular injection" refers to an injectable preparation suitable for direct injection via an injection needle into the intra-articular space within the joint capsule surrounding a joint, such as the knee. Such an intra-articular injection composition must be controlled to suit the intra-articular environment in terms of pH, osmotic pressure, viscosity, particle size, and volume, and is designed to minimize local irritation or pain upon contact with synovial fluid and joint structures. Furthermore, the composition of the present invention is characterized by being able to exert a local therapeutic effect by acting directly on tissues constituting the joint, such as cartilage, subchondral bone, and synovial membrane, upon intra-articular administration.

[0092] The above pharmaceutical composition may be characterized by being designed to be injected into the joint cavity through a superolateral approach in the affected knee joint of a patient with knee osteoarthritis. Specifically, the above pharmaceutical composition may be characterized by being designed to be injected into the joint cavity through a superolateral approach to the patella in the affected knee joint of a patient with knee osteoarthritis. Here, the superolateral approach refers to a standard intra-articular injection route in which, with the patient in a supine or seated position and the knee slightly flexed, an anatomical point based on the superior and lateral boundary of the patella is punctured to advance the injection needle into the joint cavity. This superolateral approach has the advantages of being able to secure a relatively wide space in the patellofemoral joint cavity, which is advantageous for ensuring uniform distribution of the composition within the joint cavity; it offers high safety by avoiding major neurovascular structures; and it is a route that can be repeatedly reproduced by a skilled clinician. Accordingly, the composition of the present invention can be designed with consideration of the volume, viscosity, and injectability of the formulation so as to be suitable for intra-articular injection through the upper outer region.

[0093] For example, the above pharmaceutical composition may be a single-dose formulation.

[0094] As another example, the above pharmaceutical composition may be a formulation for multiple administration or repeated administration.

[0095]

[0096] In the present invention, the knee osteoarthritis patient may be a knee osteoarthritis patient with Kellgren-Lawrence grade 2 or 3 on X-ray examination, or a knee osteoarthritis patient with cartilage defects of grade 3 or 4 according to the International Cartilage Repair Society (ICRS) criteria on magnetic resonance imaging (MRI).

[0097] In addition, the patient with knee osteoarthritis may be a patient with knee osteoarthritis with Kellgren-Lawrence grade 2 or 3 on X-ray examination, and may be a patient with knee osteoarthritis with cartilage defects of grade 3 or 4 according to International Cartilage Repair Society (ICRS) standards on magnetic resonance imaging (MRI).

[0098] In addition, the above-mentioned knee osteoarthritis patient may be a patient who complains of pain of 50 mm or more on a 100 mm VAS scale despite conservative treatment including nonsteroidal anti-inflammatory drugs, physical therapy, or exercise therapy for at least 12 weeks.

[0099] More specifically, the patient group with knee osteoarthritis defined as above in the present invention refers to patients who have moderate to severe focal cartilage defects in which cartilage damage is clearly confirmed radiographically, but who have not yet reached the stage of absolute indication for artificial joint replacement surgery. In addition, by defining patients as those with persistent pain of 50 mm or more on a 100 mm VAS scale despite receiving conventional conservative treatments such as non-steroidal anti-inflammatory drugs, physical therapy, and exercise therapy for at least 12 weeks, the patient group with intractable pain that does not respond sufficiently to existing treatments is selected.

[0100] Therefore, the pharmaceutical composition of the present invention can be used as a disease-modifying therapeutic agent to control the progression of structures such as cartilage and subchondral bone, as well as to improve pain and function, in patients with knee osteoarthritis who satisfy these clinical and radiological criteria.

[0101]

[0102] The pharmaceutical composition of the present invention may result in a decrease in the VAS pain score by 50% or more, an increase in the MOCART score by 70%, a decrease in the WORMS cartilage subscore by 40%, an increase in the IKDC score by 50% or more, a decrease in the WOMAC total score by 50% or more, or an increase in the KOOS Quality of Life (QoL) subscale by 50% or more compared to baseline at 24 weeks after administration.

[0103] The above term, "baseline point," refers to the point in time immediately before the pharmaceutical composition of the present invention is administered to a patient with knee osteoarthritis, or at the start of the trial, when the first evaluation of pain indicators (VAS), functional indicators (WOMAC, IKDC, KOOS), imaging indicators (MOCART, WORMS), biomarkers, etc., is performed. That is, the values ​​measured at the baseline point are used as reference values ​​to calculate the degree of increase or decrease (rate of change) of each indicator by comparing them with values ​​measured at follow-up points, such as at 24 weeks after administration.

[0104] For example, the pharmaceutical composition of the present invention may, at 24 weeks after administration, compared to baseline, have a VAS pain score reduced by 50% or more and / or a MOCART score increased by 70% and / or a WORMS cartilage subscore reduced by 40% and / or an IKDC score increased by 50% or more and / or a WOMAC total score reduced by 50% or more and / or a KOOS Quality of Life (QoL) subscale increased by 50% or more.

[0105] As another example, the pharmaceutical composition of the present invention may result in a decrease in the VAS pain score of 70% or more and / or an increase in the IKDC score of 80% or more and / or a decrease in the WOMAC total score of 60% or more compared to baseline at 24 weeks after administration.

[0106] The above WORMS scores, such as the VAS pain score, MOCART score, and cartilage score, IKDC score, WOMAC total score, and KOOS scores, such as the Quality of Life (QoL) subscale, may increase or decrease in one or more directions of improvement, and may include all values ​​increasing or decreasing in directions of improvement.

[0107]

[0108] At 24 weeks after administration, the above pharmaceutical composition may have a relative improvement rate of 50% or more according to OMERACT-OARSI criteria, or a proportion of responders meeting an absolute change of 20 points or more that is 50% or more of the total number of patients receiving the composition, or a proportion of responders meeting at least one scale according to KOOS that is 30% or more, or a proportion of responders according to VAS that is 60% or more of the total number of patients receiving the composition.

[0109] The term "responder" above, which simultaneously satisfies the OMERACT-OARSI criteria of a relative improvement rate of 50% or more and an absolute change of 20 points or more, refers to a subject who is judged to have shown a clinically significant level of improvement in the relevant indicator, meaning that for a specific pain or function indicator (e.g., VAS, WOMAC, IKDC, KOOS, etc.), the relative improvement rate is calculated as (24-week score - baseline score) / baseline score × 100) and this value is 50% or more, and at the same time, the absolute change (24-week score - baseline score) is at least 20 points. In other words, patients who show not merely a minor change in score, but a sufficiently large improvement (≥50%) compared to baseline and a distinct improvement (≥20 points) in absolute value can be defined as a responder according to the internationally accepted OMERACT-OARSI criteria.

[0110] The fact that the proportion of responders who simultaneously meet the OMERACT-OARSI criteria of a relative improvement rate of 50% or more and / or an absolute change of 20 points or more at 24 weeks after administration of the above pharmaceutical composition is 50% or more of the total number of patients who received the composition means that more than half of the patients show a clinically significant level of improvement with respect to the composition of the present invention when pain and functional indicators are comprehensively evaluated. Here, the OMERACT-OARSI criteria is an internationally accepted standard that defines a responder group as a case where there is a relative improvement of at least 50% or more compared to baseline and an absolute change of at least 20 points, and it is a figure that reflects significant clinical improvement felt by the patient, rather than a simple numerical change.

[0111] For example, at 24 weeks after administration of the above pharmaceutical composition, the proportion of responders meeting the OMERACT-OARSI criteria (relative improvement rate of 50% or more and absolute change of 20 points or more) may be 50% or more.

[0112] As another example, at 24 weeks after administration of the above pharmaceutical composition, the proportion of responders meeting the OMERACT-OARSI criteria (relative improvement rate of 50% or more and absolute change of 20 points or more) may be 70% or more.

[0113] In addition, the term "responder group in one or more scales based on KOOS" refers to subjects who, for one or more of the subscales of Pain, Symptoms, Activities of Daily Living (ADL), Sports and Recreation Function, and Quality of Life (QoL) evaluated by the KOOS questionnaire, have a relative improvement rate of 50% or more based on the baseline KOOS score and an absolute change of 20 points or more. That is, if the above criteria are satisfied in at least one of the five KOOS subscales, the subject can be considered as a "responder group in one or more scales based on KOOS."

[0114] For example, at 24 weeks after administration of the above pharmaceutical composition, for one or more of the subscales of Pain, Symptoms, Activities of Daily Living (ADL), Sports and Recreation Function, and Quality of Life (QoL) evaluated by the KOOS questionnaire, the relative improvement rate based on the baseline KOOS score may be 50% or more, and at the same time, the proportion of the response group with an absolute change of 20 points or more may be 30% or more.

[0115] As another example, at 24 weeks after administration of the above pharmaceutical composition, for one or more of the subscales of Pain, Symptoms, Activities of Daily Living (ADL), Sports and Recreation Function, and Quality of Life (QoL) evaluated by the KOOS questionnaire, the relative improvement rate based on the baseline KOOS score may be 50% or more, and at the same time, the proportion of the response group with an absolute change of 20 points or more may be 40% or more.

[0116] In addition, the term "responder defined by VAS" above refers to a subject whose degree of improvement, calculated using the difference between the baseline and the 24-week mark after administration regarding the VAS pain score, is greater than or equal to a predetermined standard. Specifically, a subject is defined as a responder when the relative improvement rate is -50% or more (improvement) and the absolute value changes by 20 or more. For example, when the VAS pain score at baseline is denoted as VAS_baseline and the VAS pain score at the 24-week mark as VAS_24weeks, a subject can be defined as a responder based on VAS when the value calculated by the relative improvement rate = (VAS_24weeks-VAS_baseline) / VAS_baseline×100 is -50% or more (improvement) and, at the same time, the absolute change is 20 mm or more. In other words, when the VAS pain score changes significantly in relative terms (improvement of -50% or more) and in absolute terms (20 mm or more) compared to the baseline, it can be considered as a response group based on the VAS criteria in this invention.

[0117] The fact that the proportion of the response group defined by the above VAS is 60% or more of the total number of patients who received the composition means that, even looking only at the pain intensity reported by the patients, pain was significantly reduced in a significantly high proportion of patients at 24 weeks after administration of the composition of the present invention.

[0118] For example, at 24 weeks after administration of the above pharmaceutical composition, the proportion of the response group defined by VAS may be 60% or more of the total number of patients who received the composition.

[0119] As another example, the response rate defined by VAS at 24 weeks after administration of the above pharmaceutical composition may be 80% or more of the total number of patients who received the composition.

[0120] As another example, the response rate defined by VAS at 24 weeks after administration of the above pharmaceutical composition may be 60% or more of the total number of patients who received the composition and reach 100% in the high-dose cohort.

[0121] The composition of the present invention demonstrates excellent clinical efficacy in substantially improving pain and function in patients with knee osteoarthritis by exhibiting a high response rate in both integrated indices (OMERACT-OARSI criteria) and single indices (VAS).

[0122]

[0123] For example, the above pharmaceutical composition may result in an increase in the MOCART total score compared to baseline in at least 70% of all patients at 24 weeks after administration.

[0124] For example, the pharmaceutical composition may have a proportion of patients in whom improvement compared to baseline is observed in at least one of the MOCART sub-items, such as volume fill of cartilage defect and surface of the repair tissue, at 24 weeks after administration, of 60% or more.

[0125] For example, the above pharmaceutical composition may result in a decrease in the total WORMS score compared to baseline in at least 40% of all patients at 24 weeks after administration.

[0126] For example, the above pharmaceutical composition may have a proportion of patients in whom the cartilage assessment subscore of WORMS improves compared to baseline at 24 weeks after administration of 50% or more.

[0127]

[0128] The above composition may not worsen the cartilage or subchondral bone structure compared to baseline at 24 weeks after administration, or may improve the cartilage or subchondral bone structure.

[0129] The above term, "subchondral bone," refers to bone tissue located immediately below the articular cartilage that supports the cartilage layer. Specifically, the subchondral bone is a region composed of the cartilage plate on the articular surface and the compact bone and cancellous bone beneath it, and is an important structural element that distributes and transmits body weight and mechanical loads, and reflects structural changes in osteoarthritis in the form of bone marrow edema or sclerosis.

[0130] The above subchondral bone may not deteriorate compared to baseline at 24 weeks after administering the pharmaceutical composition of the present invention, and may rather the subchondral bone structure be improved.

[0131] The improvement of the cartilage or subchondral bone structure described above refers to a change in which the morphology and signal characteristics of the subchondral bone observed on magnetic resonance imaging (MRI) are restored to a state prior to the lesion or closer to normal. Specifically, this may include the normalization or reduction of bone marrow signals, the restoration of continuity of the subchondral bone boundary, or cartilage or subchondral bone remodeling. More specifically, the normalization or reduction of bone marrow signals refers to a reduction or disappearance of high-signal regions reflecting bone marrow edema or inflammation / edema within the subchondral bone, and the restoration of continuity of the subchondral bone boundary refers to a finding in which the subchondral bone boundary line, which appeared irregular and broken, is reformed into a smooth and continuous line. Furthermore, the cartilage or subchondral bone remodeling includes a change in which the thickness, density, contour, etc., of the subchondral bone are readjusted and reorganized into a structure favorable for load distribution. These changes can serve as important structural indicators suggesting that disease-modifying effects are present not only in articular cartilage but also at the subchondral bone level in osteoarthritis.

[0132] For example, at 24 weeks after administration of the above pharmaceutical composition, the cartilage or subchondral bone structure may be maintained stably without deterioration in at least 80% of all patients, or deterioration may be delayed or the structure improved, and the cartilage or subchondral bone structure may be improved in at least 5 to 10% of all patients.

[0133] For example, the cartilage or subchondral bone remodeling may be observed in a patient receiving a high-dose composition containing 1.0 x 10 hUCB-MSCs and 60 mg of CAM.

[0134] For example, the cartilage or subchondral bone remodeling mentioned above may be derived from MOCART evaluation indicators.

[0135]

[0136] The term "biomarker" used in the present invention refers to a marker related to inflammation, cartilage synthesis, cartilage degradation, and / or other joints. Specifically, biomarkers related to inflammation are IL-1β and TNF-α, biomarkers related to cartilage synthesis are PIIANP, biomarkers related to cartilage degradation are CTX-II, and other joint-related biomarkers may be serum and urine indicators including COMP and MMP-3, but are not limited thereto.

[0137] The term IL-1β (Interleukin-1 beta) is a representative inflammatory cytokine primarily secreted by macrophages and plays a role in promoting the degradation of chondrocytes and exacerbating inflammation within the joint. In osteoarthritis, an increase in IL-1β concentration reflects increased joint inflammatory activity and cartilage destruction activity; in this invention, a decrease in IL-1β after treatment can serve as an indicator of anti-inflammatory and disease-modifying effects.

[0138] The aforementioned term, TNF-α (Tumor Necrosis Factor-alpha), is another important inflammatory cytokine that, along with IL-1β, promotes intra-articular inflammatory responses and the degradation of the cartilage matrix. In osteoarthritis, elevated TNF-α levels are associated with pain, synovial inflammation, and cartilage destruction, and a decrease in TNF-α concentration after treatment can be used as an indicator suggesting anti-inflammatory effects and pathophysiological improvement.

[0139] The above term, PIIANP (Procollagen type II N-terminal propeptide), is an N-terminal propeptide released when a precursor of type II collagen is formed, and is a biomarker that reflects cartilage matrix synthesis (regeneration). An increase in PIIANP concentration in patients with osteoarthritis (especially an increase after treatment) signifies an increase in cartilage synthesis and regeneration activity, and in the present invention, it can serve as a positive indicator for evaluating the cartilage regeneration-promoting effect.

[0140] The term CTX-II (C-terminal telopeptide of type II collagen) is a marker of cartilage degradation that is produced when type II collagen is broken down and is measured primarily in urine or blood. In osteoarthritis, an increase in CTX-II concentration indicates increased degradation of the cartilage matrix, and a decrease in CTX-II or a stable state without deterioration after treatment suggests inhibition of cartilage destruction or inhibition of disease progression.

[0141] In the above terms, serum and urine indicators, serum indicators refer to biomarkers measured in blood (serum) that reflect systemic or local pathophysiological changes, and urine indicators refer to indicators that indirectly evaluate changes in cartilage, bone, and inflammation metabolism through metabolites or degradation products measured in urine. In the present invention, serum IL-1β, TNF-α, PIIANP, CTX-II, COMP, MMP-3, etc. are utilized as serum indicators, and urine CTX-II, etc. are utilized as urine indicators, and their change patterns can be used to evaluate changes in the balance of cartilage synthesis and degradation following the administration of the composition.

[0142] The term COMP (Cartilage Oligomeric Matrix Protein) refers to a multimeric protein primarily found in the cartilage matrix, which is involved in maintaining the interaction and mechanical stability between chondrocytes and the matrix. An increase in COMP concentration in serum or synovial fluid reflects increased cartilage damage or matrix degradation, while the stabilization or decrease of COMP after treatment can serve as an indicator suggesting inhibition of the progression of cartilage structural damage.

[0143] The term MMP-3 (Matrix Metalloproteinase-3) is a type of matrix metalloproteinase that degrades the cartilage matrix (proteoglycans, collagen, etc.); its expression increases in response to inflammatory stimuli, thereby promoting cartilage destruction. In osteoarthritis, an elevation in MMP-3 concentration signifies increased cartilage-degrading activity, and a decrease or stabilization of MMP-3 concentration after treatment can be used as a biomarker suggesting inhibition of cartilage destruction mechanisms and anti-inflammatory and disease-modifying effects.

[0144] In the present invention, the composition may be one or more of the group consisting of (a) a decrease in serum IL-1β concentration, (b) a decrease in serum TNF-α concentration, (c) an increase in serum PIIANP concentration, (d) maintenance of urine CTX-II (creatinine corrected) concentration, and (e) maintenance of serum COMP and MMP-3 concentrations at 24 weeks after administration compared to baseline.

[0145] For example, a decrease in serum IL-1β and TNF-α concentrations may be predominant in the low-dose and medium-dose groups, and an increase in PIIANP concentration may be predominant in the medium-dose and high-dose groups.

[0146] For example, serum COMP and MMP-3 concentrations may remain stable without significant fluctuations from baseline for 24 weeks after administration.

[0147] The above biomarker pattern may reflect the adjustment of the intra-articular catabolic-anabolic balance toward the reduction of inflammatory cytokines and the promotion of cartilage matrix synthesis.

[0148]

[0149] The pharmaceutical composition of the present invention may further include one or more selected from the group consisting of a cryopreservative, an isotonic solution (isotonic solution), and a buffer solution.

[0150] The above term, "cryopreservative," refers to an additive used to minimize cell membrane damage, ice crystal formation, and osmotic stress during the storage and transport of living cells, such as umbilical cord blood-derived mesenchymal stem cells, under low temperature or frozen conditions. Examples of cryopreservatives may be used, such as glycerol, dimethyl sulfoxide (DMSO), or pharmaceutically acceptable combinations thereof, and are included in the cell suspension at an appropriate concentration to stably maintain the viability and function of the cells. The type and concentration of the cryopreservative may be appropriately selected and adjusted by a person skilled in the art depending on the type of cell, storage temperature, and duration.

[0151] The above term, “isotonic solution (isotonic solution),” refers to an aqueous solution formulated to have substantially the same osmotic pressure as plasma or physiological body fluids, so that the cells and chondrocyte-free matrix contained in the pharmaceutical composition of the present invention are osmotically balanced with the in vivo environment. Such an isotonic solution prevents cell swelling or shrinkage to maintain cell membrane stability and serves to stably maintain cell viability and functional activity throughout the processes of preparation, storage, transport, and administration of the composition. Examples of isotonic solutions that may be used include, but are not limited to, 0.9% sodium chloride solution (normal saline), Ringer's lactate solution, plasma electrolyte solution, and similar pharmaceutically acceptable isotonic electrolyte solutions. The isotonic solution may provide the effect of reducing local irritation, pain, and tissue damage upon administration of the composition by minimizing the osmotic pressure difference with the fluid within the joint cavity.

[0152] The term "buffer solution" above refers to an aqueous medium that enables the cells and chondrocyte-free matrix contained in the composition of the present invention to be stably dispersed and suspended, and maintains the pH and ionic strength before and after administration within a physiologically acceptable range. Examples of buffer solutions may be used, such as phosphate-buffered saline (PBS), HES solution, or other pharmaceutically acceptable electrolyte solutions, and sugars, amino acids, protein stabilizers, etc., may be additionally included as needed. The buffer solution serves to minimize the difference in osmotic pressure and pH with the intra-articular environment, thereby reducing cell viability and local irritation to the patient upon administration of the composition.

[0153]

[0154] In a specific embodiment of the present invention, through a multicenter, open-label, dose-escalation Phase 1 clinical trial involving 12 patients with K&L grade 2~3, ICRS grade 3~4 knee osteoarthritis, a low dose (2.5 x 10 7 ), medium capacity (5.0X10 7 ), high capacity (1.0X10 8 The safety and overall efficacy of a composition containing hUCB-MSC and 60 mg CAM when administered as a single intra-articular dose were confirmed.

[0155] In addition, by analyzing the dose-response relationship, it was confirmed that the improvement in pain and functional indicators such as VAS, WOMAC, IKDC, and KOOS in the intermediate-to-high dose administration groups was significantly superior compared to the low dose group; in particular, through comparative examples, even with the same composition, the low dose (2.5X10 7 It has been shown that improvements in pain, function, and structural indicators are somewhat limited in the dosage range specified in the present invention (e.g., 5.0 x 10 7 ~1.0×10 8 It was confirmed that ) is an important factor in exerting disease control effects.

[0156] Furthermore, structural improvement and disease control effects at the cartilage and subchondral bone levels were presented through the maintenance (delayed deterioration) and improvement of MOCART and WORMS scores and K&L grade, and biomarker patterns suggesting a reconfiguration of catabolic-anabolic balance in the direction of reducing inflammation and promoting cartilage matrix synthesis were presented through the analysis of biomarkers such as IL-1β, TNF-α, PIIANP, CTX-II, COMP, and MMP-3.

[0157] Through this, it can be seen that the pharmaceutical composition of the present invention possesses excellent safety and efficacy with only a single intra-articular administration, and has efficacy as a DMOAD therapeutic agent for controlling knee osteoarthritis, encompassing clinical indicators, imaging indicators, and biomarker indicators within a specific dose range compared to low doses.

[0158]

[0159] Unless otherwise indicated in this specification, a composition according to the present invention may include, in addition to the materials described in the description of the invention or the claims, "pharmaceutically acceptable carriers, excipients, or diluents" in addition to the materials described herein.

[0160] In the present invention, the term "pharmaceuticalally acceptable" means a sufficient amount to produce a therapeutic effect and not causing side effects, and can be easily determined by a person skilled in the art based on factors well known in the medical field, such as the type of disease, age, weight, health, gender, sensitivity of the individual to the drug, route of administration, method of administration, frequency of administration, duration of treatment, and drugs used in combination or concurrently.

[0161] In the present invention, the term “pharmaceuticalally acceptable carrier” typically comprises a liquid or non-liquid basis of a pharmaceutical composition. If the pharmaceutical composition is provided in liquid form, the carrier is typically pyrogen-free water; isotonic saline or a buffer (aqueous) solution, e.g., a buffer solution such as phosphate, citric acid, etc. The injection buffer may be hypertonic, isotonic, or hypotonic to a specific reference medium. That is, the buffer may have a high, equal, or low salt content to a specific reference medium, and preferably, an appropriate salt concentration may be used within a range that does not induce cell damage due to osmotic pressure or other concentration effects. The reference medium is blood, lymph, cytoplasmic liquid, or other body fluid, or a liquid occurring in an “in vivo” method, such as a general buffer or liquid that can be used as a reference medium in an “in vitro” method. Such general buffers or liquids are known to a person skilled in the art.

[0162] The above pharmaceutically acceptable carriers include, but are not limited to, those commonly used in the art, such as 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, and mineral oil.

[0163] In addition, the pharmaceutical composition of the present invention may include diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants, as well as other pharmaceutically acceptable additives.

[0164] In addition, the pharmaceutical composition of the present invention may further include a cell cryopreservation agent.

[0165]

[0166] The pharmaceutical composition of the present invention may further include one or more active ingredients exhibiting the same or similar medicinal effects in addition to the fusion formulation of the present invention.

[0167] The pharmaceutical composition according to the present invention may be included in a "pharmaceuticalally effective amount." The pharmaceutically effective amount is used in a conventional sense and refers to an amount that is sufficient to produce a preventive or therapeutic effect against osteoarthritis upon administration of the composition, while ensuring that side effects are within an acceptable range. The effective amount may be appropriately adjusted by a person skilled in the art, taking into account the patient's age, weight, severity of the disease, medical history, concomitant medications, etc.

[0168] Since the effective dosage of the above pharmaceutical composition is determined by considering various factors such as the method of formulation, route of administration, and number of treatments, as well as the patient's age, weight, health status, gender, severity of the disease, diet, and excretion rate, a person with ordinary knowledge in the art will be able to determine an appropriate effective dosage of the composition of the present invention by taking these points into account.

[0169]

[0170] The pharmaceutical composition of the present invention can be formulated in the form of a sterile injectable solution according to conventional methods. In addition, it can be prepared in the form of a liquid or a suspension, among which it can be prepared as a suspension.

[0171] Specifically, the pharmaceutical composition of the present invention may be intended for parenteral administration (e.g., intra-articular injection). In the present invention, the term "parenteral administration" refers to administration via intravitreal injection, retro-orbital injection, subretinal injection, subcutaneous, intramuscular, intravenous, or intraperitoneal via a tube, excluding oral administration; specifically, it refers to a pharmaceutical composition intended for intra-articular injection. The "intra-articular administration" refers to a method of administration in which the pharmaceutical composition is directly injected into the knee joint cavity using an injection needle, and the composition may be injected by an orthopedic specialist through a supererolateral approach of the knee joint. After administration, it is desirable to observe the patient for a certain period of time to check for adverse reactions such as injection site reactions, worsening of pain, or edema.

[0172] In parenteral administration, the composition for local administration may be anhydrous or aqueous depending on the clinical prescription. As non-aqueous solvents and suspending agents, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. may be used.

[0173] The umbilical cord blood-derived mesenchymal stem cells and chondrocyte-free matrix of the present invention can be administered via a single intra-articular injection, and in particular, can be injected into the joint cavity of the knee joint to simultaneously improve pain, function, and imaging structural indicators in patients with knee osteoarthritis. The pharmaceutical composition can provide significant clinical and structural improvements during a typical observation period with just a single administration, and may be repeated after a certain period at the clinician's discretion if necessary.

[0174]

[0175] In the present invention, the term "treatment" refers to any act of administering the above composition to a patient with already diagnosed osteoarthritis to improve pain, functional impairment, and radiological abnormalities, or to inhibit the progression of the disease.

[0176] In the present invention, "individual" refers to a mammal including humans, dogs, monkeys, cats, rabbits, etc., and specifically, may be an individual with osteoarthritis, more specifically, an adult mammal with knee osteoarthritis.

[0177]

[0178] The above description may be applied to other embodiments or other aspects of the present invention, but is not limited thereto.

[0179]

[0180] Another aspect of the present invention for solving the above problem is a formulation for intra-articular administration comprising a combination of umbilical cord blood-derived mesenchymal stem cells and chondrocyte-free matrix.

[0181] For example, the above preparation may be for single administration.

[0182] As another example, the above preparation may be for repeated administration.

[0183] The above terms, "cord-derived mesenchymal stem cells," "chondrocellular matrix," "intra-articular," "preparation," etc., are as described above.

[0184] For example, the pharmaceutical composition of the present invention is about 2.5 x 10 per dose. 7 Up to 1.0X10 8 It may contain canine umbilical cord blood-derived mesenchymal stem cells and about 30 mg to 120 mg, more specifically 60 mg, of chondrocyte-free matrix. More specifically, the number of cells is 2.5 x 10 7 , 5.0×10 7 or 1.0X10 8 It may be a dog, and the content of the above-mentioned cartilage acellular matrix may be 60 mg.

[0185] As another example, the above pharmaceutical composition may be formulated such that the total injection volume is about 2 mL to 5 mL, more specifically about 2.25 mL, 3.0 mL, or 4.5 mL. For example, a cord blood-derived mesenchymal stem cell suspension (e.g., a cell preparation suspended in CS10 medium) and a chondrocyte-free matrix suspension (e.g., a 60 mg / 1.5 mL physiological saline suspension) may be mixed and provided as a single formulation injectable into the joint cavity.

[0186]

[0187] Another aspect of the present invention for solving the above problem is a method for treating osteoarthritis comprising the step of administering the pharmaceutical composition to an individual.

[0188] The above terms, "pharmaceutical composition," "individual," "administration," "osteoarthritis," "treatment," etc., are as described above.

[0189]

[0190] Another aspect of the present invention for solving the above problem is a method for promoting cartilage or subchondral bone remodeling, comprising the step of administering the pharmaceutical composition to an individual.

[0191] The above terms, "pharmaceutical composition," "individual," "administration," "cartilage or subchondral bone remodeling," "treatment," etc., are as described above.

[0192]

[0193] Another aspect of the present invention for solving the above problem is a kit for manufacturing an intra-articular administration agent for disease control of knee osteoarthritis, comprising: a first vial containing umbilical cord blood-derived mesenchymal stem cells; and a second vial containing a chondrocellular matrix.

[0194] The above terms, "umbilical cord blood-derived mesenchymal stem cells," "chondrocellular matrix," "knee osteoarthritis," "for disease control," "intra-articular administration," etc., are as described above.

[0195] The term "kit" above refers to a set of two or more components provided together in a package for the purpose of preparing, manufacturing, and administering an intra-articular agent for the control of knee osteoarthritis. For example, the kit may include a configuration provided in a single package unit comprising a first vial containing umbilical cord blood-derived mesenchymal stem cells, a second vial containing chondrocellular matrix, a buffer solution for dilution or suspension as needed, a syringe and needle, and instructions for use. The user may use the kit to prepare an intra-articular injectable composition by mixing the contents of the first vial and the second vial immediately before administration, and then administer it to a patient.

[0196] The first vial may contain umbilical cord blood-derived mesenchymal stem cells in the form of a cell suspension containing a cryopreservation agent and a buffer solution, or in a form usable after freezing and thawing, and the second vial may contain a chondrocyte-free matrix in the form of a suspension or freeze-dried, and may be configured so that the two preparations can be mixed in a single syringe immediately before administration and injected into the joint cavity.

[0197] If necessary, the above kit may additionally include a dilution buffer solution, a syringe, a needle, instructions for use, etc., and can be administered to the affected knee joint of a patient with knee osteoarthritis via a conventional intra-articular injection route including a supererolateral approach.

[0198] For example, the above kit is i) 2.5X10 7 ii) a first vial containing hUCB-MSC and a second vial containing 60 mg of CAM, ii) 5.0X10 7 A first vial containing 1 / 2 hUCB-MSCs and a second vial containing 60 mg of CAM, or iii) 1.0×10 8 It may be a first vial containing hUCB-MSC and a second vial containing 60 mg of CAM.

[0199] For example, the above first vial and second vial are mixed immediately before administration, 5.0×10 7 Up to 1.0X10 8 It may be configured to administer a 2.0 to 4.5 mL suspension containing 2 hUCB-MSCs and 50 to 70 mg of CAM.

[0200] For example, the above kit may be characterized by being configured for a single administration into the knee joint cavity of a patient with knee osteoarthritis.

[0201] As another example, the above kit may be characterized by being configured for repeated administration into the knee joint cavity of a patient with knee osteoarthritis.

[0202]

[0203] Thus, the disease-modifying therapeutic composition for osteoarthritis and the composition for intra-articular administration, which are composed of a combination of umbilical cord blood-derived mesenchymal stem cells and a chondrocellular matrix according to the present invention, provide a disease-modifying osteoarthritis treatment strategy that is differentiated from existing osteoarthritis treatments by providing clinically significant pain and functional improvement accompanied by structural improvement at the cartilage and sub-bone levels, which was not sufficiently confirmed in conventional umbilical cord blood-derived mesenchymal stem cell monotherapy, and by possessing an acceptable safety profile.

[0204] Due to these technical features, the present invention can provide a new clinical composition, its use, and a treatment method that are clearly distinguished from the prior art regarding compositions for treating degenerative diseases using umbilical cord blood-derived mesenchymal stem cells.

[0205]

[0206] The present invention will be explained in more detail below through examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples. Meanwhile, technical matters not described in this specification can be fully understood and easily implemented by a person skilled in the art who is proficient in the technical field of this application or a similar technical field.

[0207]

[0208] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention may be applied to other descriptions and embodiments thereof. That is, 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 provided below.

[0209] Expressions such as “comprising” as used in this specification should be understood as open-ended terms implying the possibility of including other experimental examples, unless specifically stated otherwise in the phrase or sentence containing such expression.

[0210] Terms and words used in the description and claims of the present invention shall not be interpreted as being limited to their ordinary or dictionary meanings, but shall be interpreted in a meaning and concept consistent with the technical spirit of the present invention, based on the principle that the inventor can appropriately define the concept of the terms to best describe his invention.

[0211]

[0212] Example 1: Design of K0701 Phase 1 Clinical Trial and Confirmation of Overall Safety and Efficacy

[0213] In order to confirm whether the umbilical cord blood-derived mesenchymal stem cell (hUCB-MSC) and chondrocyte-free matrix (CAM) combination preparation (OSCA) of the present invention is clinically significant for osteoarthritis, and to confirm the appropriate dosage and method of administration, as well as whether it has safety and efficacy with only a single administration, a Phase 1 clinical trial of K0701 was designed for patients with knee osteoarthritis.

[0214] Specifically, the clinical trial design is explained in detail below, divided into sections, based on FIG. 1, a diagram schematically illustrating the mechanism of action of the cord blood-derived mesenchymal stem cell / chondrocyte-free matrix composite (OSCA) of the present invention; FIG. 2, a diagram schematically illustrating the Phase 1 clinical trial design of the clinical trial (K0701) of the present invention; and FIG. 3, a test protocol flowchart showing the visit schedule and evaluation items of the clinical trial (K0701) of the present invention.

[0215]

[0216] 1-1. Study Design and Subjects

[0217] To evaluate the safety, tolerability, and exploratory efficacy of OSCA, an injectable combination of a cell therapy containing umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) and cartilage acellular matrix (CAM), a multicenter, open-label, dose-escalation Phase 1 clinical trial (K0701) was conducted in patients with knee osteoarthritis.

[0218] First, subjects were selected from adults (aged 19 or older) diagnosed with knee osteoarthritis corresponding to radiographic K&L grade 2-3 or MRI ICRS grade 3 or 4 at the time of screening, limited to patients with a 100 mm VAS pain score ≥ 50 mm and whose symptoms did not improve despite conservative treatment (medication, physical therapy, etc.) for 12 weeks or more. Specifically, the subject selection and exclusion criteria applied to the clinical trial (K0701) of the present invention were carried out according to the information presented at https: / clinicaltrials.gov / study / NCT05944627.

[0219] More specifically, a total of 12 participants were enrolled in 3 dosage cohorts (low dosage 2.5×10⁻¹⁰). 7 cells + CAM 60 mg, intermediate dose 5.0 x 10 7 cells + CAM 60 mg, high dose 1.0×10 8 They were sequentially assigned to cells + CAM 60 mg). The low and medium doses each consisted of 3 subjects, and the high dose consisted of 6 subjects, following a 3+3 dose escalation design. Finally, 24 weeks of follow-up were completed for 10 subjects. Table 1 below (Demographic and baseline characteristics of the three cohorts) shows the baseline characteristics of subjects who participated in the clinical trial (K0701) of the present invention by cohort.

[0220]

[0221]

[0222] 1-2. Administration Method, Specimens, and Visit Schedule

[0223] On the day of administration, frozen osiramestrocel (hUCB-MSC formulation, Kangstem Co., Ltd.) and CAM formulation were thawed and mixed using a single syringe. The injection site was prepared aseptically, and a single intra-articular administration was performed by an orthopedic specialist via a supererolateral approach to the relevant knee joint. Adverse reactions (such as injection site reactions) were monitored for 30 minutes after administration.

[0224] Urine and blood specimens were collected at baseline and at 1, 4, 12, and 24 weeks after injection. The first morning urine in midstream form was stored at -70°C. Blood specimens were left at room temperature for 30 minutes, then centrifuged at 3000 rpm for 10 minutes, and the resulting serum was stored at -70°C or below until analysis.

[0225]

[0226] Meanwhile, study visits were conducted on Day 1 (administration day) and at weeks 1, 4, 8, 12, 16, 20, and 24 after administration. At each visit, safety (physical examination, vital signs, laboratory tests, TEAE / ADR / SAE) and pain and functional assessments (IKDC, KOOS, WOMAC, VAS) were performed. Structural evaluations via MRI and X-ray (MOCART 2.0, WORMS, K&L grade) were performed at baseline and at week 24. The specific details of the structural / evaluation schedule are illustrated in Figures 2 and 3. Additionally, each protocol can be referenced at https: / / clinicaltrials.gov / study / NCT05944627 (Ann.rheum. Dis.(1957), 16, 494., OsteoArthritis and Cartilage (2004) 12, 389-399., Original Article Knee Surg Relat Res 2013;25(3):106-111., Journal of Orthopaedic&sports Plysical Therapy March 9, 2025., Arthritis Care & Research Vol. 63, No. S11, November 2011, pp S240-S252, http: / / oncology.thelancet.com Vol 7 November 2006, Cartilage 2021, Vol. 13(Suppl 1) 571S-587S, OsteoArthritis and Cartilage (See (2004) 12, 177-190, semjnars in Arthritis and Rheumetism, Vol 18. No 4, Suppi 2 (May). 1989: pp 14-17, etc.)

[0227] For reference, celecoxib 200 mg / day (200 mg once daily or 100 mg twice daily) was permitted as a rescue drug during the study period, and administration of the rescue drug was to be discontinued for at least 5 days immediately prior to the efficacy evaluation. In addition, NSAIDs, muscle relaxants, systemic steroids, and other intra-articular injection therapies were designated as prohibited drugs to minimize confusion in the evaluation.

[0228]

[0229] 1-3. Evaluation Variables and Statistical Analysis

[0230] The primary endpoints were the occurrence of dose-limiting toxicity (DLT) and the determination of the maximum tolerated dose (MTD). Safety and tolerability were assessed across all cohorts based on the occurrence of DLT and the frequency of treatment-emergent adverse events (TEAEs), adverse drug reactions (ADRs), and serious adverse events (SAEs). The MTD was determined by monitoring the frequency and severity of DLT within the dose escalation design, and adverse events were classified and graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI-CTCAE ver. 5.0).

[0231] Exploratory efficacy was confirmed for secondary endpoints based on changes in pain and function scores over 24 weeks following OSCA administration. For this purpose, the International Knee Documentation Committee (IKDC), Knee Injury and Osteoarthritis Outcome Score (KOOS), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and Visual Analogue Scale (VAS) indices were used. Higher IKDC and KOOS scores indicate better clinical outcomes, while lower WOMAC and VAS scores indicate symptom relief.

[0232] In addition, further clinical efficacy analysis was performed by classifying the responders and non-responders based on the degree of improvement in pain or function. Here, the responders were defined as those who showed a relative improvement of 50% or more and an absolute improvement of 20 points or more compared to baseline.

[0233] In addition, to confirm the potential disease-modifying effects of OSCA, changes in joint structure were assessed using MRI and radiography. MRI evaluations were performed using the Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART) 2.0 score and the Whole-Organ Magnetic Resonance Imaging Score (WORMS), while radiographic evaluations were performed using the K&L grading system. MOCART and WORMS evaluations were conducted using a 3T MR system (Achieva and Ingenia, Philips Medical Systems, Eindhoven, The Netherlands) at baseline and at 24 weeks, respectively, prior to the single IA OSCA injection. Detailed MRI sequence parameters are presented in Figure 3.

[0234] Image evaluation MOCART, WORMS, and K&L grades were interpreted as meaning structural improvement of the joint when grades were maintained (delayed deterioration) or improved. All MRI and X-ray images were uploaded to a centralized clinical trial image management system (Trial Informatics, Seoul, South Korea), and blinded analysis was performed by two independent radiologists. In cases where the judgments of the two evaluators did not agree, a third independent judgment radiologist reviewed the results of the first and second readings and selected the most valid result without performing a separate re-reading.

[0235] Additionally, an exploratory biomarker analysis was performed over 24 weeks to investigate the biological effects of OSCA on cartilage degradation and inflammation.

[0236] For statistical analysis, continuous variables were presented as mean and standard deviation, while categorical variables were presented as frequency and percentage. All analyses of evaluation variables were based solely on observed data, and no imputation was performed for missing values. After testing for normality, the Kruskal-Wallis test or one-way ANOVA was used for inter-cohort comparisons of continuous variables, while the Mann-Whitney U test or Tukey's HSD was used for post hoc pairwise comparisons as necessary.

[0237] Categorical variables were analyzed using the Pearson chi-square test or Fisher's exact test, and changes from baseline at 24 weeks within the cohort were evaluated using the Wilcoxon signed-rank test.

[0238] A linear mixed-effect model (LMM) was used to evaluate differences between cohorts during the 24-week follow-up period. In this model, patients were included as random effects, while cohort, visit time, and interactions between them were included as fixed effects. To identify significant differences between cohorts, post-hoc pairwise comparisons were additionally performed.

[0239] Data analysis was performed using SAS Version 9.4 (SAS Institute, Cary, NC, USA), and a p-value of less than 0.05 in a two-sided test was determined to be statistically significant.

[0240]

[0241] 1-4. Safety and Overall Efficacy Results

[0242] Table 2 below summarizes the adverse reactions and drug adverse reactions observed after administration of the test drug (OSCA) of the present invention.

[0243] OutcomeLow-dose(N = 3)n (%) [Event]Mid-dose(N = 3)n (%) [Event]High-dose(N = 6)n (%) [Event]All doses(N = 12)n (%) [Event]TEAEs0 (0) [0]1 (33.33) [1]2 (33.33) [3]3 (25.00) [4]ADRs0 (0) [0]0 (0) [0]1 (16.67) [1]1 (8.33) [1]SAEs0 (0) [0]0 (0) [0]1 (16.67) [1]1 (8.33) [1]SADRS0 (0) [0]0 (0) [0]0 (0) [0]0 (0) [0]DLT occurrence0 (0) [0]0 (0) [0]0 (0) [0]0 (0) [0]TEAEs leading to death0 (0) [0]0 (0) [0]0 (0) [0]0 (0) [0]TEAEs by SeverityGrade 10 (0) [0]0 (0) [0]0 (0) [0]0 (0) [0]Grade 20 (0) [0]1 (33.33) [1]2 (33.33) [2]3 (25.00) [3]Grade 30 (0) [0]0 (0) [0]1 (16.66) [1]1 (8.33) [1]Grade 40 (0) [0]0 (0) [0]0 (0) [0]0 (0) [0]Grade 50 (0) [0]0 (0) [0]0 (0) [0]0 (0) [0]TEAEs by Relationship to IPRelated0 (0) [0]0 (0) [0]1 (16.67) [1]1 (8.33) [1]Not related0 (0) [0]1 (33.33) [1]2 (33.33) [2]3 (25.00) [3]Results of TEAEsRecovered / resolved0 (0) [0]1 (33.33) [1]2 (33.33) [2]3 (25.00) [3]Recovering / resolving0 (0) [0]0 (0) [0]1 (16.66) [1]1 (8.33) [1]Not recovered / not resolved0 (0) [0]0 (0) [0]0 (0) [0]0 (0) [0]Death0 (0) [0]0 (0) [0]0 (0) [0]0 (0) [0]Unknown0 (0) [0]0 (0) [0]0 (0) [0]0 (0) [0].

[0244] *TEAEs (Treatment-Emergent Adverse Events) -> Adverse events occurring during treatment (all adverse events newly occurring or worsening after the start of treatment) / ADRs (Adverse Drug Reactions) -> Adverse drug reactions (adverse events judged to be causally related to the administered drug) / SAEs (Serious Adverse Events) -> Serious adverse events (including serious medical events such as death, life-threatening conditions, hospitalization / prolonged hospitalization, disability / functional impairment, congenital abnormalities, etc.) / SADRs (Serious Adverse Drug Reactions) -> Serious adverse drug reactions (serious adverse events related to drug administration) / DLT (Dose Limiting Toxicity) -> Dose-limiting toxicity (serious toxicity occurring when a dose exceeding the prescribed dose is administered in a clinical trial; criteria for establishing the Maximum Tolerable Date (MTD)) / IP (Investigational Product) -> Investigational Product (experimental drugs used in clinical trials, including new drugs or comparators)

[0245] As a result, as can be seen in Table 2 above, no DLT was identified in any cohort, and no TEAEs leading to death were reported. Grade 2-3 TEAEs occurred in 3 out of 12 patients (25.0%), and the only ADR assessed as being related to the investigational drug was one case of knee edema (8.3%) observed in the high-dose cohort, which recovered within 6 weeks without sequelae. One case of acute transverse myelitis (8.3%) was reported as an SAE, but it was determined to be unrelated to the investigational drug.

[0246] More specifically, at 24 weeks after administration, the intermediate and high-dose cohorts (Cohorts 2 and 3) showed statistically significant improvement in all pain and functional indicators, including VAS, WOMAC, IKDC, and KOOS, compared to the low-dose cohort (p < 0.05, LMM and post-hoc analysis). In particular, VAS decreased by up to approximately 91.3% and IKDC increased by up to 102.7%, confirming significant improvement in joint pain and function.

[0247] In addition, radiologically, as confirmed in Figure 4, MOCART scores increased in 70% of all patients, while WORMS criteria showed a decrease (improvement) in 40%, and K&L grades were maintained stably in 80%. Furthermore, as shown in Figure 5, MOCART results indicated that volume fill in cartilage defect areas improved in 60% of patients, subchondral bone structure remained stable or delayed deterioration in 80%, and improved in 10%. Notably, subchondral bone remodeling was observed only in the high-dose cohort, confirming superior efficacy within a specific dosage range.

[0248]

[0249] Example 2: Confirmation of symptom and functional improvement following administration of low, medium, and high doses of OSCA

[0250] Next, to determine at which dosage range OSCA provides superior pain and functional improvement effects, we specified low, medium, and high dosage ranges and directly compared their effects.

[0251]

[0252] 2-1. Subjects and Administration

[0253] Under the same test design as in Example 1 above, a low dose (2.5X10 7cells), medium dose (5.0 x 10⁻⁶ 7 cells), high dose (1.0 x 10⁻⁶ 8 Three, three, and six subjects were assigned to cells, respectively. At this time, it was confirmed that there were no statistically significant differences among the three groups in baseline characteristics (age, sex, BMI, duration of disease, baseline VAS / WOMAC / IKDC / KOOS) among the subjects (see Table 1 above).

[0254] All subjects received a single intra-articular injection and followed the same visit and evaluation schedule for 24 weeks. The conditions, such as the type of rescue medication used and restrictions on simultaneous administration, were the same as those in Example 1 above.

[0255]

[0256] 2-2. Improvement of Pain and Functional Indicators and Dose-Response Analysis

[0257] First, changes in pain and functional indicators (VAS, WOMAC, IKDC, KOOS) over time and by dose are shown in Figure 4. Specifically, Figure 4 is a graph showing changes in pain and functional indicators (VAS, WOMAC, IKDC, KOOS, etc.) by dose cohort up to 24 weeks after a single intra-articular administration of the composition of the present invention to a patient with knee osteoarthritis.

[0258] As a result, as can be seen in Figure 4, low, medium, and high doses were all effective in pain and functional indicators. Among them, the medium and high dose groups showed greater reduction in pain and improvement in function and quality of life compared to baseline compared to the low dose group. In particular, it was confirmed that the IKDC score in the high dose group increased by more than 100% compared to baseline at 24 weeks, despite being a single administration. Through this, it was confirmed that there was a significant clinical improvement effect in the medium and high doses compared to the low dose.

[0259] In addition, the rate of change in major indicators for each cohort at the 24-week mark is as shown in Table 3 below. Specifically, Tables 3 to 10 below show the changes in clinical indicators such as VAS, WOMAC, IKDC, and KOOS by cohort at the 24-week mark after administration of the test drug (OSCA) of the present invention.

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269] As a result, as can be seen in Tables 3 to 10 above, regarding the rate of change in IKDC scores (compared to baseline), it was confirmed that the low dose increased by approximately 15.3%, the medium dose by approximately 89.7%, and the high dose by approximately 102.7%. Although it was confirmed that there was an increase in all three doses, among them, the medium and high doses showed an effect of increasing by approximately 74% to 95% compared to the low dose, confirming that they had a significantly superior effect compared to the low dose.

[0270] In addition, regarding the rate of change in WOMAC scores (compared to base, decrease is an improvement), it was confirmed that the low dose decreased by approximately 4.7%, the medium dose by approximately 60.5%, and the high dose by approximately 76.2%, confirming that all three doses were effective. Among them, it was confirmed that the medium and high doses showed superior improvement effects compared to the low dose.

[0271] In addition, regarding the rate of change in VAS pain scores (compared to baseline, reduction was improved), it was confirmed that all three doses showed improvement effects by showing a decrease of approximately 14.6% at the low dose, approximately 78.9% at the medium dose, and approximately 91.3% at the high dose. Among them, similar to the IKDC and WOMAC scores mentioned above, it can be seen that the medium and high doses show a significantly better pain improvement effect compared to the low dose.

[0272] Similarly, regarding all three subscales of KOOS (pain, symptoms, daily living, sports and leisure, quality of life), it was confirmed that all three doses showed improvement effects, but the intermediate and high dose cohorts consistently showed greater improvement compared to the low dose. In post-hoc pairwise comparisons, it was also confirmed that all three doses were effective, but the intermediate and high doses had superior effects compared to the low dose, thus confirming that there was a somewhat significant difference between the intermediate and high doses and the low dose in the overall results.

[0273] The above results suggest that while OSCA is effective for treating osteoarthritis at low, medium, and high doses, the medium and high doses demonstrate significantly superior disease-controlling effects compared to the low dose.

[0274]

[0275] 2-3. Response Group Analysis

[0276] Next, the responder analysis according to OMERACT-OARSI standards was verified, and the results are shown in Table 11 below.

[0277] For reference, Table 11 below is the result of a post-hoc analysis showing the number of patients who achieved significant pain or functional improvement at the 24-week mark in the clinical trial (K0701) of the present invention.

[0278] (Low-dose, N = 3)(Mid-dose, N = 3)(High-dose, N = 4)P- valueVAS0.027Responder (Yes)0 (0.0)2 (66.7)4 (100.0)Non-responder (No)3 (100.0)1 (33.3)0 (0.0)WOMAC0.115Responder (Yes)0 (0.0)2 (66.7)3 (75.0)Non-responder (No)3 (100.0)1 (33.3)1 (25.0)IKDC0.129Responder (Yes)0 (0.0)1 (33.3)3 (75.0)Non-responder (No)3 (100.0)2 (66.7)1 (25.0)KOOS (Pain)0.217Responder (Yes)0 (0.0)2 (66.7)2 (50.0)Non-responder (No)3 (100.0)1 (33.3)2 (50.0)KOOS (Symptoms)0.197Responder (Yes)0 (0.0)2 (66.7)1 (25.0)Non-responder (No)3 (100.0)1 (33.3)3 (75.0)KOOS(Activities of daily living)0.217Responder (Yes)0 (0.0)2 (66.7)2 (50.0)Non-responder (No)3 (100.0)1 (33.3)2 (50.0)KOOS(Sports and recreation function)0.356Responder (Yes)0 (0.0)1 (33.3)2 (50.0)Non-responder (No)3 (100.0)2 (66.7)2 (50.0)KOOS (Quality of life)0.115Responder (Yes)0 (0.0)2 (66.7)3 (75.0)Non-responder (No)3 (100.0)1 (33.3)1 (25.0)

[0279] * In the table above, the high improvement group (responders) is defined as a relative improvement rate of 50% or more from baseline and an absolute score change of 20 points or more. Abbreviations are defined as follows: VAS: Visual Analog Scale / WOMAC: University of Western Ontario and McMaster Osteoarthritis Index / IKDC: International Knee Documentation Committee Index / KOOS: Knee Injury and Osteoarthritis Outcome Score

[0280] As a result, as can be seen in Table 11 above, the VAS response rate at 24 weeks was confirmed to have reached 100% in Cohort 3 (high dose), and a high response rate compared to the low dose was also confirmed in the medium dose cohort (Table 11).

[0281] In addition, based on the KOOS subscales, it was confirmed that the overall figures were Pain 40%, Symptoms 30%, ADL 40%, Sports and Recreation Function 30%, and Quality of Life (QoL) 50%. When examined by dose, unlike the low dose where the response group was 0% across all KOOS subscales, it was found that in the medium and high doses, the KOOS response group was better in the medium dose than in the high dose (pain, symptoms, ADL) or in the high dose than in the medium dose (Sports and Recreation Function, Quality of Life (QoL)). Through this, it was confirmed that the proportion of the response group was also higher in the medium and high dose cohorts compared to the low dose in terms of KOOS scores.

[0282] These results demonstrate that despite being a single intra-articular administration, all three doses show significant improvement and therapeutic effects, and in particular, OSCA induces significant pain reduction and functional improvement at intermediate to high doses, and regarding the specific dose range (e.g., ≥5.0X10) in the claim of the treatment method of the present invention 7 , preferably 5.0X107 - 1.0X10 8 This result indicates that cells have a very superior effect in reducing pain and improving function compared to low doses.

[0283]

[0284] Example 3: Structural Disease Control Effect - Cartilage Defect Filling and Cartilage / Subchondral Bone Remodeling

[0285] Next, it was confirmed whether the composition of the present invention has a disease-modifying (DMOAD) effect by verifying structural changes at the level of cartilage and subchondral bone identified through MRI and radiographic evaluation after OSCA administration.

[0286]

[0287] 3-1. Video Evaluation Method

[0288] For the same patient group as in Example 1 above, MOCART 2.0 and WORMS were assessed using 3T MRI at baseline and 24 weeks, and K&L grades were interpreted using standard radiographs. Specific MRI imaging conditions (sequence, coil, position, etc.) are as shown in Tables 12 and 13 below. At this time, the interpretation was performed by two independent radiologists in a blinded manner, and the final score was determined by having a third radiologist intervene in the event of a discrepancy.

[0289]

[0290]

[0291]

[0292] Meanwhile, the MOCART score was analyzed based on defect filling, tissue integration, surface, and structure of the cartilage defect site, and WORMS was used as a semi-quantitative indicator for pre-knee structures such as cartilage, subchondral bone marrow lesions, osteophytes, subchondral cysts, and synovitis.

[0293]

[0294] 3-2. Structural Changes in Articular Cartilage and Subchondral Bone

[0295] First, an analysis of changes in articular cartilage and subchondral bone structure was performed. As can be seen in Figure 5A, MOCART scores increased in 70% (7 / 10) of the total patients at 24 weeks, which suggests defect filling and cartilage recovery in the cartilage defect area. In 60% (6 / 10) of the patients, volume fill in the cartilage defect area was analyzed to have improved.

[0296] The subchondral bone structure remained stable without deterioration or deterioration was delayed in 80% (8 / 10) of patients, and improvement was confirmed in 10% (1 / 10) of patients. In particular, unlike other doses, subchondral bone remodeling was confirmed in the high-dose cohort (Cohort 3), and improvement in bone marrow signaling and structural remodeling were observed in the patients (Fig. 5).

[0297] Next, as can be seen in Figure 5B, the total WORMS score showed no statistically significant change based on the overall average of the patient group, but showed a decrease (improvement) in 40% (4 / 10). Looking only at the cartilage evaluation item within WORMS, improvement was confirmed in 50% (5 / 10), indicating that structural recovery at the cartilage level was partially achieved. On the other hand, the osteophyte score showed an increasing trend in 70% (7 / 10), confirming the possibility of osteogenic proliferation caused by growth factors such as stem cell-derived TGF-β and BMP-2.

[0298] Finally, it was confirmed that the K&L grade was maintained at 80% (8 / 10) and deteriorated by one grade at 20% (2 / 10), but overall, the structural preservation effect was dominant.

[0299] These results support the claim that, from a clinical perspective, the OSCA of the present invention is a candidate for a disease-modifying drug (DMOAD) that not only relieves simple symptoms but also partially fills and restores cartilage defects (MOCART, defect filling), maintains or partially remodels the subchondral bone structure, and, particularly at high doses, exhibits excellent remodeling effects and ensures the stability of the joint structure for at least 24 weeks.

[0300] In particular, the fact that subchondral bone remodeling was confirmed at high doses can be interpreted as a result indicating that the aforementioned high-dose range is a therapeutically effective dose sufficient to induce subchondral bone remodeling.

[0301]

[0302] 3-3. Confirmation of Clinical and Structural Improvement Upon OSCA Administration

[0303] Clinical and structural improvements were confirmed upon administration of low to high doses of OSCA.

[0304]

[0305] 3-3-1.

[0306] Structural results of the cohort

[0307] Tables 14 to 20 below show the longitudinal changes in MOCART and WORMS scores before and after administration of the test drug (OSCA) of the present invention (baseline and 24 weeks).

[0308] As can be seen from the results in Tables 14 to 20 below, improvements in cartilage defect filling were confirmed in MOCART and WORMS analyses ranging from low to high doses. However, while some patients in the low-dose cohort showed minimal improvement in cartilage defect filling, the overall degree and frequency of structural improvement were lower compared to the intermediate and high-dose cohorts, confirming that the intermediate and high doses demonstrated superior efficacy compared to the low dose (Tables 14 to 20 / MOCART - Includes Total MOCART Score, Volume fill of cartilage defect, Integration into the adjacent cartilage, Surface of the repair tissue, Structure of the repair tissue, Signal intensity of the repair tissue, Bony defect or bony overgrowth, and Subchondral change / WORMS - Includes Total WORMS score, Cartilage Assessment, Marrow Abnormality, Bone cysts, Bone Attrition, Osteophyte, Menisci, Ligaments, Synovitis, Loose Body, and Periarticular Cyst / Bursa). Subchondral bone remodeling was not observed in the low-dose cohort and was confirmed only in the high-dose cohort.

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315]

[0316]

[0317] The above results demonstrate that while OSCA composed of the same composition exhibits therapeutic effects for osteoarthritis at various doses, the clinical symptoms and structural improvement effects can vary significantly depending on the number of administered cells (dose). This suggests that intermediate to high doses exhibit clearly superior efficacy compared to simple hUCB-MSC therapy or low-dose OSCA administration. In particular, compared to low doses, intermediate to high doses suggest that the treatment can be utilized as a disease-modifying therapeutic agent for osteoarthritis, going beyond mere symptom relief.

[0318]

[0319] Example 4: Subchondral bone remodeling and bone marrow signal changes after a single intra-articular administration of OSCA

[0320] Next, the effects of a single intra-articular administration of the cord blood-derived mesenchymal stem cell-chondrocyte-acellular matrix composite (OSCA) of the present invention on subchondral bone structure and bone marrow signaling in patients with knee osteoarthritis were confirmed.

[0321] Specifically, in the same Phase 1 clinical trial described in Example 1 above, subchondral bone structure and bone marrow abnormalities were analyzed using 3T MRI at baseline and at 24 weeks after administration. Here, the MRI evaluation focused on subchondral bone-related items (e.g., subchondral changes, marrow abnormality) in MOCART 2.0 and WORMS scores, and was interpreted in a blinded manner by two independent radiologists.

[0322] As a result, as can be seen in Figure 5, structural stability of the subchondral bone was maintained in 9 out of 10 evaluable subjects (90%); among them, 80% (8 / 10) maintained a stable state without structural progression, and 10% (1 / 10) showed improvement in the subchondral bone structure. Subchondral bone remodeling was observed in a high-dose cohort (1.0×10⁻¹⁰). 8 This was confirmed in the high-dose cohort (cells + CAM 60 mg), and in 1 out of 4 evaluable subjects (25%) in the high-dose cohort, the subchondral bone marrow signal was improved and accompanied by structural remodeling (sorting of structural boundaries, reduction of bone marrow edema, etc.) (Fig. 5).

[0323] In one high-dose subject, bone marrow edema in the medial femoral condyle region was observed on baseline MRI, but normalization of bone marrow signals and restoration of the continuity of the subchondral bone boundary were confirmed in the same region at 24 weeks after administration (Fig. 6 (C), (D)). This suggests that structural recovery or remodeling may occur at the subchondral bone level along with the regulation of the joint microenvironment after OSCA administration.

[0324] The results of the above examples show that the composition according to the present invention induces not only defect filling and cartilage recovery in cartilage defect sites, but also stabilization of the subchondral bone structure and remodeling in some patients.

[0325]

[0326] Example 5: Confirmation of joint structure improvement through MOCART and WORMS score patterns

[0327] Next, the pattern of change in MOCART and WORMS scores after a single intra-articular administration of the composition (OSCA) of the present invention was examined to quantitatively confirm the improvement in joint structure in patients with knee osteoarthritis.

[0328] Specifically, for the same patient group as in Example 1 above, MOCART 2.0 and WORMS were analyzed at baseline and at 24 weeks after administration. MOCART consists of 7 items (total 100 points), including volume fill, integration with surrounding cartilage, surface and structure of regenerated tissue, signal intensity, bone defect or bony hyperplasia, and subchondral bone changes; a higher score indicates that the cartilage condition is closer to normal. WORMS semi-quantitatively evaluates the entire knee structure, including cartilage, bone marrow abnormalities, bone cysts, bone loss, osteophytes, menisci, ligaments, synovitis, intra-articular loose bodies, and peri-cysts; a lower score indicates a structure closer to normal.

[0329] As a result, as can be seen in Figure 5, at 24 weeks, the MOCART total score increased from baseline in 70% (7 / 10) of the total patients, indicating that regeneration and filling of the cartilage defect site had progressed in a significant number of subjects (Figure 5(A)). In particular, significant improvement was frequently observed in the volume fill and regenerative tissue surface sub-items of MOCART. Improvement in volume fill of the cartilage defect site was confirmed in 60% (6 / 10) of the total patients, and MRI images showed smoothing of the cartilage surface and partial or nearly complete filling of the defect.

[0330] Although there was no statistically significant difference in the WORMS total score between baseline and 24 weeks based on the overall average, the WORMS total score decreased (improved) in 40% (4 / 10) of the subjects (Fig. 5(B)). Among the WORMS sub-items, the score corresponding to cartilage assessment showed improvement in 50% (5 / 10) of the patients, suggesting that significant structural improvement at the cartilage level was observed in some subjects. On the other hand, the osteophyte score showed a worsening trend in 70% (7 / 10) of the subjects, but this is interpreted as a phenomenon related to the activity of regenerative growth factors after OSCA administration.

[0331]

[0332] In the case of the marrow abnormality subscore, most subjects showed a tendency toward stability or slight improvement without deterioration, which supports the effect of preserving joint structure at the subchondral bone level. According to Tables 19 through 26 described earlier, the average score for the marrow abnormality item decreased or remained almost unchanged at 24 weeks, and in the LMM analysis, differences between groups were not statistically significant, but no trend of deterioration over time was observed.

[0333] This example demonstrates that the composition of the present invention can induce preservation and partial recovery of joint structure for at least 24 weeks through improvement in the MOCART score reflecting cartilage regeneration (increase from 70%) and improvement in some WORMS subscores including cartilage and bone marrow evaluation items (40–50%) after a single OSCA administration.

[0334]

[0335] Example 6: Changes in Inflammation and Cartilage Metabolism-Related Biomarkers After Single OSCA Administration

[0336] Finally, after administering the cord blood-derived mesenchymal stem cell-chondrocyte-acellular matrix composite (OSCA) of the present invention as a single intra-articular injection, changes in serum and urine biomarkers related to inflammation and cartilage metabolism were confirmed.

[0337] Specifically, in the same Phase 1 clinical trial as in Example 1 above, serum and urine samples were collected at baseline (before administration) and at 1, 4, 12, and 24 weeks after administration. CTX-I, CTX-II, TNF-α, IL-1β, COMP, MMP-3, and PIIANP were measured in serum, while CTX-I and CTX-II were measured in urine. CTX-I and CTX-II in urine were analyzed after being corrected for creatinine concentration. Each analysis was performed in the central laboratory using ELISA or multiplex immunoassay, and quality control was conducted based on calibration curve accuracy and recovery rate (% recovery).

[0338] As a result, as can be seen in Figure 7, serum IL-1β and TNF-α, inflammation-related markers, showed an overall decreasing trend in the low-dose and intermediate-dose groups during the 24 weeks after OSCA administration. PIIANP, a marker reflecting cartilage matrix synthesis, showed an increasing trend in the intermediate and high-dose groups, while a decreasing trend was observed in the low-dose group.

[0339] In the case of CTX-II, which reflects cartilage degradation, urinary CTX-II remained stable without significant change over 24 weeks across all dose groups, while serum CTX-II showed a slight increasing trend. COMP and MMP-3 also remained stable without significant fluctuation across all dose groups (Fig. 6). These results suggest that no apparent acceleration of cartilage degradation occurred during the study period (24 weeks).

[0340] To summarize the above results, after administration of the composition of the present invention to osteoarthritis patients, a decreasing trend in inflammation-related markers (IL-β, TNF-α) was observed in the low-to-medium dose groups, confirming the alleviation of intra-articular inflammatory responses. Furthermore, regarding the cartilage synthesis-related marker (PIIANP), an increasing trend was observed in the medium-to-high dose groups, whereas a decreasing trend was observed in the low dose group. This indicates the possibility that cartilage matrix synthesis is promoted at doses above a certain level, specifically medium and high doses, rather than at low doses. Moreover, in the analysis of the cartilage degradation-related marker (CTX-II), urinary CTX-II remained stable, and serum CTX-II showed only slight fluctuations without significant changes, confirming that excessive promotion of cartilage degradation was not observed in the short term. Finally, other structure and inflammation-related markers (COMP, MMP-3, etc.) generally showed a stable pattern.

[0341]

[0342] In conclusion, the above results provide biological evidence that the OSCA of the present invention inhibits inflammatory cytokines (IL-1β, TNF-α) in the intra-articular microenvironment and, above a certain dose, readjusts the catabolic-anabolic balance in a direction that promotes cartilage matrix synthesis (PIIANP), and particularly readjusts it even more at intermediate and high doses compared to low doses.

[0343]

[0344] From the foregoing description, those skilled in the art to which the present invention pertains will understand that the present invention may be implemented in other specific forms without altering its technical concept or essential features. In this regard, the embodiments described above should be understood as illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as including all modifications or variations derived from the meaning and scope of the claims set forth below and their equivalents, rather than from the detailed description above.

Claims

1. A pharmaceutical composition for treating osteoarthritis comprising human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) and cartilage acellular matrix (CAM).

2. In Paragraph 1, The above hUCB-MSC is 2.5 x 10 per single administration 7 Up to 1.0X10 8 A pharmaceutical composition for treating osteoarthritis containing a number of cells.

3. In Paragraph 2, The above hUCB-MSC is 5.0X10 7 Up to 1.0X10 8 A pharmaceutical composition for treating osteoarthritis containing a number of cells.

4. In Paragraph 1, A pharmaceutical composition for treating osteoarthritis, wherein the above CAM is included in an amount of 40 to 80 mg per single administration.

5. In Paragraph 4, A pharmaceutical composition for treating osteoarthritis, wherein the above CAM is included in an amount of 50 to 70 mg per single administration.

6. In Paragraph 1, A pharmaceutical composition for treating osteoarthritis, wherein the above-mentioned osteoarthritis is knee osteoarthritis.

7. In Paragraph 1, A pharmaceutical composition for treating osteoarthritis, wherein the above composition is formulated in the form of a suspension having a total dosage of 2.0 mL to 4.5 mL.

8. In Paragraph 7, The above composition is, i) 2.5X10 7 ii) a 2.25 mL formulation containing hUCB-MSC and 60 mg of CAM, 5.0X10 7 A 3.0 mL formulation containing hUCB-MSC and 60 mg of CAM, or iii) 1.0×10 8 A pharmaceutical composition for disease modification of osteoarthritis, comprising a 4.5 mL formulation containing 60 mg of hUCB-MSC and 60 mg of CAM.

9. In Paragraph 1, A pharmaceutical composition for treating osteoarthritis, characterized in that the above composition further comprises one or more selected from the group consisting of a cryopreservative, an isotonic solution, and a buffer solution.

10. In Paragraph 1, A pharmaceutical composition for treating osteoarthritis, characterized by mixing an hUCB-MSC preparation and a CAM preparation in a single syringe immediately before administration.

11. In Paragraph 1, The above composition is a pharmaceutical composition for treating osteoarthritis, which is for intra-articular injection.

12. In Paragraph 11, A pharmaceutical composition for treating osteoarthritis, characterized in that the above composition is designed to be injected into the joint cavity through a supererolateral approach in the affected knee joint of a patient with knee osteoarthritis.

13. In Paragraph 12, A pharmaceutical composition for treating osteoarthritis, wherein the knee osteoarthritis patient is a knee osteoarthritis patient with Kellgren-Lawrence grade 2 or 3 on X-ray examination, or a knee osteoarthritis patient having localized cartilage defects of grade 3 or 4 according to International Cartilage Repair Society (ICRS) standards on magnetic resonance imaging (MRI).

14. In Paragraph 12, A pharmaceutical composition for treating osteoarthritis, wherein the patient with knee osteoarthritis is a patient who complains of pain of 50 mm or more on a 100 mm VAS scale despite conservative treatment including non-steroidal anti-inflammatory drugs, physical therapy, or exercise therapy for at least 12 weeks.

15. In Paragraph 1, The above composition, compared to baseline at 24 weeks after administration, (a) If the VAS pain score decreases by 50% or more, or (b) If the MOCART score increases by 70% or more, or (c) If the WORMS cartilage subscore decreases by 40% or more, or (d) If the IKDC score increases by 50% or more, or (e) If the total WOMAC score decreases by 50% or more, or (f) A pharmaceutical composition for treating osteoarthritis that increases the KOOS Quality of Life (QoL) subscale by 50% or more.

16. In Paragraph 1, At 24 weeks after administration of the above composition, if the proportion of responders meeting the OMERACT-OARSI criteria of a relative improvement rate of 50% or more and an absolute change score of 20 points or more is 50% or more, or If the response rate is 30% or higher on one or more scales based on KOOS, or A pharmaceutical composition for treating osteoarthritis having a response rate of 60% or more based on VAS.

17. In Paragraph 1, The above composition is a pharmaceutical composition for treating osteoarthritis characterized by maintaining the cartilage or subchondral bone structure relative to baseline, delaying deterioration, or improving the subchondral bone structure at 24 weeks after administration.

18. In Paragraph 17, A pharmaceutical composition for treating osteoarthritis, wherein the improvement of the subchondral bone structure described above includes normalization or reduction of bone marrow signals, restoration of continuity of the subchondral bone boundary, or cartilage or subchondral bone remodeling.

19. In Paragraph 1, At 24 weeks after administration of the above composition, compared to baseline, (a) Decrease in serum IL-1β concentration, (b) Decrease in serum TNF-α concentration, (c) Increase in serum PIIANP concentration, (d) Maintain urine CTX-II (creatinine corrected) concentration, or (e) A pharmaceutical composition for treating osteoarthritis having the characteristic of maintaining serum COMP and MMP-3 concentrations.

20. A method for treating osteoarthritis comprising the step of administering a pharmaceutical composition of any one of claims 1 to 19 to an individual.

21. A method for promoting cartilage or subchondral bone remodeling comprising the step of administering a pharmaceutical composition of any one of claims 1 to 19 to an individual.

22. A kit for manufacturing an intra-articular administration agent for the treatment of knee osteoarthritis, comprising: a first vial containing umbilical cord blood-derived mesenchymal stem cells; and a second vial containing chondrocellular matrix.

23. In Paragraph 22, The above kit is i) 2.5X10 7 ii) a first vial containing hUCB-MSC and a second vial containing 60 mg of CAM, ii) 5.0X10 7 A first vial containing 1 / 2 hUCB-MSCs and a second vial containing 60 mg of CAM, or iii) 1.0×10 8 A kit comprising a first vial containing 10 hUCB-MSCs and a second vial containing 60 mg of CAM.

24. In Paragraph 22, The above first vial and second vial are mixed immediately before administration, 5.0X10 7 Up to 1.0X10 8 A kit configured to administer a 2.0 to 4.5 mL suspension containing 2 hUCB-MSCs and 50 to 70 mg of CAM.

25. In Paragraph 22, A kit characterized by being configured for a single administration into the knee joint cavity of a patient with knee osteoarthritis.

Images