Inhibitor of expression of bone formation-related factors or calcification-related factors in extraskeletal tissues

Abstract

Provided is an inhibitor of the expression of osteogenesis-related factors or calcification-related factors in extraskeletal tissues. The inhibitor is an inhibitor of the expression of osteogenesis-related factors in extraskeletal tissues, characterized by containing phytic acid. The osteogenesis-related factor is any one of ALPL, RUNX2, BGLAP, etc. The inhibitor is an inhibitor of the expression of calcification-related factors in extraskeletal tissues, characterized by containing phytic acid. The calcification-related factor is any one of SLC20A1, SLC20A2, ENPP1, ALPL, SPP1, etc. Ectopic ossification and ectopic calcification can be suppressed by intravenous administration.

Description

[Technical Field]

[0001] The present invention relates to an agent for suppressing the expression of osteogenesis-related factors or calcification-related factors in extraskeletal tissues. [Background technology]

[0002] Bone-forming cells include osteoblasts, osteocytes, and osteoclasts. Osteoblasts are bone-forming cells differentiated from mesenchymal stem cells, and differentiate in stages from stem cells. In cultured osteoblastic cell lines, marker genes that indicate the cell differentiation stage show diverse patterns. For example, during the early proliferation stage, cells express CCND1 and HDAC, which are proteins that control the cell cycle. However, differentiation phenotypes are expressed in a time-dependent manner, with COL1A1 being expressed early in differentiation. Subsequently, as cells mature, the expression pattern is shown in the order of FN1, ALPL, IBSP, and BGLAP. Among these markers, COL1A1 and ALPL are expressed independently of the cell cycle, while BGLAP is expressed after the end of cell division.

[0003] Other osteoblast markers include transcription factors such as RUNX2, SPP1, MSX2, DLX5, TWIST1, and JUN. RUNX2 promotes the transcription of the BGLAP and SPP1 genes by binding to upstream cis-elements, suggesting that RUNX2 plays an important role in the production of bone matrix proteins.

[0004] However, when the bone formation-related factors described above are expressed in extraskeletal tissues, ectopic ossification, a phenomenon in which pathological bone formation occurs in areas where bone tissue does not normally form, can occur, separate from normal skeletal formation. Furthermore, calcification is a general term for calcium deposition, and the phenomenon of pathological calcification occurring in areas where calcification does not normally occur is called ectopic calcification. It has been shown that at least some of this ectopic calcification occurs through the same molecular mechanism as bone formation (Non-Patent Document 1). Tissues in which ectopic ossification or ectopic calcification occurs can cause various disorders. For example, in joint areas, it is accompanied by limited range of motion, resulting in a significant decrease in quality of life (QOL). In blood vessels, it is associated with cardiovascular events and life prognosis.

[0005] Patent Document 1 describes a regulator of osteoblast differentiation that focuses on the fact that NRF2 can suppress Bglap transcriptional activity in a Runx2-dependent manner, and that the negative regulation of bone and cartilage differentiation by nrf2 may be due to the inhibition of Runx2 by NRF2. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Re-tabled publication No. 2006 / 129881 [Non-patent literature]

[0007] [Non-Patent Document 1] Circ.Res.89,1147-1154, 2001; Semin. Nephrol. 24, 61-68, 2004 Summary of the Invention [Problem to be solved by the invention]

[0008] However, the substance described in Patent Document 1 does not sufficiently suppress the expression of bone formation-related factors or calcification-related factors.

[0009] The present invention has been made in view of the above problems, and an object of the present invention is to provide a novel agent for suppressing the expression of osteogenesis-related factors or calcification-related factors in extraskeletal tissues. [Means for solving the problem]

[0010] The agent for suppressing the expression of bone formation-related factors or calcification-related factors in extraskeletal tissues according to the present invention is characterized by containing phytic acid. [Effects of the Invention]

[0011] According to the present invention, the expression of osteogenesis-related factors or calcification-related factors in extraskeletal tissues can be appropriately suppressed. [Brief explanation of the drawings]

[0012] [Figure 1] Figure 1 shows the effect of phytic acid administration in an ectopic ossification model. (A) is a micro-CT image of an ectopic ossification model showing ossified tissue at the site of implantation of atelocollagen impregnated with BMP-2 into mouse muscle tissue. (B) shows the dose-dependent reduction in the amount of ossified tissue due to phytic acid. [Figure 2] Figure 1 shows the effect of phytic acid administration on a calcification model of mouse aortic organ cultures under 2.6 mM phosphate (Pi) loading. (A) is a photograph of a thoracic aortic organ culture excised from a mouse, (B) is a micro-CT image showing the effect of phytic acid administration on calcification of mouse aortas cultured under Pi loading, and (C) is a graph showing the effect of phytic acid administration on calcification of mouse aortic organs cultured under Pi loading. [Figure 3] FIG. 1 is a photograph of von Kossa staining showing the effect of phytic acid administration in a mouse aortic organ calcification model under a 2.6 mM phosphate (Pi) loading environment. [Figure 4] FIG. 1 is a photograph of immunostaining of the osteoblast marker ALPL in a mouse aortic organ calcification model in a 2.6 mM phosphate (Pi) loaded environment, where phytic acid was administered. [Figure 5] This figure shows the gene expression status when phytic acid was administered to a mouse aortic organ calcification model under a 2.6 mM phosphate (Pi) load environment. (A) is Alpl, (B) is Runx2, (C) is Bglap, and (D) is Slc20a1. [Figure 6]This figure shows the inhibitory effect of phytic acid on warfarin-induced arterial calcification. (A) is a photograph of a blood vessel from a negative control group administered with vitamin K1 alone. (B) is a photograph of a blood vessel that had become calcified after warfarin administration. (C) is a photograph of a state in which calcification was inhibited when phytic acid was further administered at a dose of 2 mg / 50-120 g body weight / day. [Figure 7] FIG. 1 shows serum calcium concentrations in mice administered phytic acid intravenously. [Figure 8] FIG. 1 shows serum inorganic phosphate concentrations in mice administered phytic acid intravenously. [Figure 9] Photographs of the aortic arch of Enpp1 mutant mice, in which (A) is vehicle administration, (B) is 0.4 mg / body / day sodium phytate administration, and (C) is 0.04 mg / body / day sodium phytate administration. [Figure 10] This figure shows the results of bone morphometry of the proximal tibia in Enpp1 mutant mice, where (A) is cortical bone mass, (B) is cancellous bone mass, (C) is cortical bone density, and (D) is cancellous bone density. [Figure 11] FIG. 1 shows the results of serum samples administered with 0.4 mg / body / day of sodium phytate and the vehicle, where (A) is the serum zinc concentration and (B) is the serum iron concentration. DETAILED DESCRIPTION OF THE INVENTION

[0013] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, these embodiments are intended to facilitate understanding of the principles of the present invention, and the scope of the present invention is not limited to the following embodiments. Other embodiments in which a person skilled in the art appropriately replaces the configuration of the following embodiments are also included in the scope of the present invention.

[0014] Phytic acid is a substance with the following structural formula. It is a component found in large amounts in unrefined grains and beans and exhibits a strong chelating effect on minerals. It is approved as a food additive as an acidulant and pH adjuster, and is also widely disclosed as a functional ingredient due to its potential for various physiological functions, including antioxidant effects.

[0015] [ka]

[0016] However, it has not been known that phytic acid has an effect of suppressing the expression of bone formation-related factors or calcification-related factors in extraskeletal tissues.

[0017] The agent for suppressing the expression of bone formation-related factors or calcification-related factors in extraskeletal tissues according to the present invention is characterized by containing phytic acid.

[0018] Phytic acid can also be used in the form of a phytate, and although not particularly limited, examples thereof include calcium salt, magnesium salt, and zinc salt of phytic acid.

[0019] The inhibitor of the expression of bone formation-related factors or calcification-related factors in extraskeletal tissues according to the present invention contains a pharmaceutically effective amount of phytic acid, and can be made into oral preparations such as tablets, granules, fine granules, capsules, etc., various liquid preparations suitable for oral administration, or parenteral preparations such as injections and suppositories.

[0020] Among parenteral preparations, injectable preparations are prepared in the form of, for example, a solution, emulsion, or suspension, and are made isotonic with blood. Preparations in the form of a liquid, emulsion, or suspension are prepared using, for example, an aqueous medium, ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, or polyoxyethylene sorbitan fatty acid ester. Examples of aqueous media include water or media containing water. Sterile water is used as water. Examples of media containing water include physiological saline, PBS (phosphate buffered saline), and lactated Ringer's solution.

[0021] In the injectable preparation, the content of phytic acid or a salt thereof is not particularly limited, but is, for example, 1 μg / mL to 10 mg / mL, and preferably 100 μg / mL to 1 mg / mL.

[0022] Additives commonly used in the art can be used in injectable formulations. Examples of additives include isotonicity agents, stabilizers, buffers, preservatives, chelating agents, antioxidants, and solubilizing agents. Examples of isotonicity agents include sugars such as glucose, sorbitol, and mannitol, sodium chloride, glycerin, propylene glycol, and polyethylene glycol. Examples of stabilizers include sodium sulfite. Examples of buffers include borate buffers, phosphate buffers, citrate buffers, tartrate buffers, and acetate buffers. Examples of preservatives include parahydroxybenzoates, benzyl alcohol, chlorocresol, phenethyl alcohol, and benzethonium chloride. Examples of chelating agents include sodium edetate and sodium citrate. Examples of antioxidants include sodium sulfite, sodium bisulfite, sodium ascorbate, and sodium thiosulfate. Examples of solubilizing agents include dextran, polyvinylpyrrolidone, sodium benzoate, ethylenediamine, salicylic acid amide, nicotinic acid amide, polyoxyethylene hydrogenated castor oil derivatives, and the like.

[0023] The injectable preparation may contain a pH adjuster. The pH adjuster may be an acid or a base. Specific examples of acids include ascorbic acid, hydrochloric acid, gluconic acid, acetic acid, lactic acid, boric acid, phosphoric acid, sulfuric acid, tartaric acid, and citric acid. Examples of bases include potassium hydroxide, calcium hydroxide, sodium hydroxide, magnesium hydroxide, monoethanolamine, diethanolamine, and triethanolamine.

[0024] The bone formation-related factors are not particularly limited, but examples thereof include ALPL, BGLAP, RUNX2, SP7 (Osterix), DLX5, and the like.

[0025] Bone mineralization-related factors are not particularly limited, but examples thereof include SLC20A1, SLC20A2, ENPP1, ALPL, and SPP1.

[0026] In the present invention, the expression of bone formation-related factors or calcification-related factors in extraskeletal tissues is suppressed by the action of phytic acid, thereby making it possible to suppress, for example, ectopic ossification and ectopic calcification.

[0027] In this regard, some have suggested that ectopic ossification is suppressed by the chelating action of phytic acid; however, the present inventors have discovered that the key mechanism by which phytic acid suppresses ectopic ossification is not its chelating action, but rather its suppression of the expression of bone formation-related factors or calcification-related factors.

[0028] Heterotopic ossification mainly includes posttraumatic ossification, tumor-induced ossification, and ossification in neurological conditions. Posttraumatic ossification is ossification that occurs at the attachment site of the medial collateral ligament of the medial femoral condyle after major trauma or repeated minor trauma. Tumor-induced ossification is soft tissue ossification that occurs in parosteal osteosarcoma, soft tissue osteosarcoma, intramuscular hemangioma, etc. Neurological ossification is ossification that occurs in paraplegia or quadriplegia due to spinal cord injury, or in the paralyzed limbs in hemiplegia due to stroke. Fibrodysplasia ossificans progressiva is a disease characterized by systemic skeletal deformities and soft tissue ossification.

[0029] Ectopic calcification mainly includes metastatic calcification, dystrophic calcification, and idiopathic calcinosis. Metastatic calcification is calcification that occurs due to long-term hypercalcemia and / or hyperphosphatemia. Dystrophic calcification is calcification that occurs in soft tissues even in the absence of disorders of calcium or phosphorus metabolism. Idiopathic calcinosis includes calcific tendonitis and bursitis, chondrocalcinosis, focal calcinosis, and generalized calcinosis, which occur most frequently in the greater tuberosity of the humerus, especially at the insertion of the supraspinatus muscle and deltoid muscle, and the lateral epicondyle of the knee joint.

[0030] Ectopic calcification occurs through some of the same molecular mechanisms as ectopic ossification, and is a phenomenon in which bone-like tissue forms in areas where bone tissue does not normally exist, such as muscles, fascia, ligaments, joint capsules, blood vessels, etc. Specific diseases that fall under this category include designated intractable diseases such as kidney disease, vascular calcification associated with dialysis, vascular calcification due to arteriosclerosis, generalized arterial calcification in infancy (GACI), fibrodysplasia ossificans progressiva (FOP), ossification of the posterior longitudinal ligament (OPLL), and ossification of the ligamentum flavum.

[0031] The preventive and / or therapeutic agent for ectopic ossification or ectopic calcification caused by the expression of bone formation-related factors or calcification-related factors according to this embodiment is characterized by containing phytic acid. The disease causing ectopic ossification is not particularly limited, and examples thereof include FOP, OPLL, and ossification of the ligamentum flavum, as listed in the specific examples above. FOP is a genetic disease characterized by congenital malformations of the thumb and progressive ectopic ossification. FOP may also be accompanied by a genetic mutation, such as a mutation in ACVR1. FOP causes extra-articular stiffness of the major joints in the axial and appendicular skeletons or thoracic fusion, resulting in severe disability or respiratory failure. The disease causing ectopic calcification is not particularly limited, and examples thereof include vascular calcification associated with kidney disease or dialysis, vascular calcification due to arteriosclerosis, and GACI, as listed in the specific examples above.

[0032] As used herein, "prevention" includes suppressing and delaying the onset of a disease, and includes not only prevention before the onset of the disease, but also prevention of the recurrence of the disease after treatment. On the other hand, "treatment" includes curing symptoms, ameliorating symptoms, and suppressing the progression of symptoms. The dosage and administration of the agent for preventing and / or treating ectopic ossification or ectopic calcification caused by the expression of bone formation-related factors or calcification-related factors according to this embodiment can be varied as appropriate, but for example, the amount of active ingredient is about 0.1 to about 2000 mg / kg / day, preferably about 1 to 200 mg / kg / day, and this amount can be administered once a day or in 2 to 3 divided doses. [Example]

[0033] (1) Example 1: Effect of intravenous administration of phytic acid on BMP-2-dependent intramuscular ossification model Atelocollagen impregnated with BMP-2 was implanted into the gluteus maximus of 4-week-old ddY male mice, creating a heterotopic ossification model in which ossification was observed at the implant site two weeks after implantation. As shown in Figure 1(A) (Vehicle), micro-CT images confirmed the formation of ossified tissue in the heterotopic ossification model.

[0034] Sodium phytate was continuously administered into the right jugular vein using an osmotic pump (Alzet) at 0.08 mg / body / day, 0.4 mg / body / day, or 2 mg / body / day, and the effect on heterotopic ossification was examined. As shown in Figure 1(A) and (B), phytic acid reduced the amount of ossified tissue in a dose-dependent manner. In Figure 1(B), the vertical axis represents bone mass (mm ) measured by micro-CT. 3 )

[0035] (2) Example 2: Effect of phytic acid administration on calcification of mouse aortic organ cultures during phosphate loading Thoracic aortas were excised from 4-week-old C57BL / 6 male mice and cut into 2-5 mm lengths (Figure 2(A)). These were placed on culture dishes and cultured in DMEM (high glucose) containing 15% fetal bovine serum at 37°C for 10 days. To create a phosphate-loaded environment, 2.6 mM phosphate (Pi) was added to the culture medium.

[0036] After 10 days of culture, mineralized tissue was observed in the mouse aorta. Mineralization of bone matrix and other matrices is initiated by matrix vesicles produced and secreted by osteoblasts present on the bone surface (matrix vesicular mineralization). The calcium phosphate crystal clusters formed within the matrix vesicles exhibit ribbon- or needle-like shapes. When they break through the vesicle membrane and are exposed to the outside world, they form spherical aggregates (mineralized nodules). These then come into contact with surrounding collagen fibrils, causing the mineralization of the collagen fibers. Osteoblasts are thus the cells responsible for bone formation and induce mineralization.

[0037] In this mouse aorta organ culture model with phosphate loading, 1 μM, 3 μM, or 10 μM phytic acid was added to the culture medium, and after 10 days of culture, the presence or absence of calcified tissue was observed. As shown in Figure 2(B) and (C), the amount of calcified tissue was reduced in a dose-dependent manner in micro-CT images of phytic acid. In Figure 2(C), the vertical axis represents the amount of calcified tissue (mm 3 )

[0038] (3) Example 3: Calcification tissue staining of mouse aortic organ cultures under phosphate loading For the mouse aorta organ culture model under phosphate loading conditions described in Example 2 above, after the completion of the culture, the tissue was fixed in 4% paraformaldehyde in PBS to prepare paraffin specimens. Five-micrometer sections were prepared and subjected to von Kossa staining. von Kossa staining is a staining method used to detect deposited calcium (calcium salts) in histological examinations, and uses silver nitrate to stain calcium phosphate salts, calcium carbonate salts, and other salts black.

[0039] As shown in Figure 3, the area stained black decreased in a dose-dependent manner with phytic acid, indicating a decrease in the amount of calcified tissue. The scale bar in Figure 3 is 25 μm.

[0040] (4) Example 4 Histological staining of osteoblast markers Using the paraffin sections from the mouse aorta organ culture model under phosphate loading conditions described in Example 2, immunostaining for the osteoblast (bone formation) marker ALP was attempted. ALP (alkaline phosphatase) is a glycoprotein present in the cell membrane. ALP specifically present in bone tissue is called BAP, which is present in the cell membrane and binds to the membrane via phosphatidylinositol. Blood BAP levels increase during periods of enhanced osteoblast function and bone formation activity, making ALP an osteoblast marker. Immunostaining was performed using standard methods with anti-ALP antibody and Alexa594-labeled secondary antibody, followed by observation under a fluorescence microscope. DAPI was used as a counterstain. As shown in Figure 4, the osteoblast marker ALP stained red when treated with solvent alone during phosphate loading. On the other hand, the red staining area decreased in a dose-dependent manner with phytic acid, indicating the inhibition of osteoblast-like cell formation. The scale bar in Figure 4 is 75 μm.

[0041] (5) Example 5 Expression of osteoblast marker genes The expression of osteoblast (bone formation) marker genes was examined by RT-PCR in the mouse aorta organ culture model under a phosphate-loaded environment described in Example 2 above. After completion of the culture, RNA was extracted according to standard methods, and cDNA was prepared. Gene expression was then quantified by real-time PCR. The expression of AlpI, Runx2, and Bglap, bone formation-related factors, was examined. Here, the AlpI gene encodes the tissue-nonspecific isozyme of alkaline phosphatase (TNSALP). Runx2 is a transcription factor essential for osteoblast differentiation and late chondrocyte differentiation. Bglap is expressed at high levels during the late stage of differentiation, when cultured cells calcify, and is therefore a marker for confirming the differentiation orientation of mesenchymal stem cells into osteoblasts.

[0042] We investigated the expression of Slc20a1 as a calcification-related factor. SLC20A1 is a type III phosphate transporter known to play an important role in calcification. Increased blood phosphate concentration induces calcification of vascular smooth muscle, leading to atherosclerosis, primarily involving calcification of the tunica media. This is thought to be due to increased extracellular phosphate concentration, which increases the intracellular influx of phosphate via SLC20A1 in vascular smooth muscle cells, inducing the expression of osteoblast differentiation-related proteins, leading to differentiation into osteoblast-like cells and calcification.

[0043] As shown in Figure 5(A), (B), and (C), the expression of bone formation-related factors was suppressed in the presence of phytic acid. Also, as shown in Figure 5(D), the expression of mineralization-related factors was suppressed in the presence of phytic acid.

[0044] (6) Example 6: Inhibition of warfarin-induced arterial calcification Vitamin K inhibitors, such as warfarin, inhibit the conversion of matrix GIa protein (MGP) to Gla, a calcification inhibitor, and suppress its function. In this example, we investigated the inhibitory effect of phytic acid on warfarin-induced arterial calcification.

[0045] At 15 days of age, newborn SD rats were switched from a standard diet to a high-phosphate diet containing 1.2 wt% phosphate and 0.6 wt% calcium, providing a high phosphate load to the mothers. At 20 days of age, oral administration of vitamin K1 (1.5 mg / 100 g body weight / day) was initiated. At 21 days of age, sodium phytate (0.4 and 2 mg / 50-120 g body weight / day) was continuously administered via the right jugular vein using an osmotic pump. After the osmotic pump was fitted, the rats were weaned and continued to receive the oral vitamin K1 and high-phosphate diet. From the following day, 22 days of age, warfarin (15 mg / 100 g body weight) was administered subcutaneously every 12 hours. On the 12th day after the start of warfarin administration, the arteries (from the aortic arch to the femoral artery) were isolated, fixed in 4% paraformaldehyde in PBS, and stained with Alizarin Red S, which stains calcified tissue red. As a result, no calcification was observed in the group that did not receive warfarin, as shown in Figure 6(A), but calcification was observed in the group that received warfarin, as shown in Figure 6(B). On the other hand, when phytic acid was also administered in addition to warfarin, aortic calcification was sufficiently suppressed.

[0046] (7) Example 7: Changes in serum calcium (Ca) and phosphate (inorganic phosphate, Pi) concentrations following intravenous administration of phytic acid Phytic acid was administered intravenously to 4-week-old male ddY mice using an osmotic pump for one week, and changes in blood calcium levels were measured. Phytic acid was administered at doses of 0.08 mg / body / day, 0.4 mg / body / day, and 2 mg / body / day. Calcium levels were measured using a Wako Calcium E-Test. The results are shown in Figure 7. As shown in Figure 7, serum calcium levels were unaffected by phytic acid. Similarly, serum Pi levels were measured using a Wako Phospha C-Test. The results are shown in Figure 8. As shown in Figure 8, phytic acid did not affect serum Pi levels.

[0047] (8) Example 8: Calcification inhibitory effect in Enpp1 mutant mice Enpp1 is the gene responsible for ossification of the posterior longitudinal ligament, and many infantile generalized arterial calcifications are associated with biallelic mutations of this gene. Enpp1 mutant neonatal mice (C57BL / 6J-Enpp1 asj Mother mice (GrsrJ / GrsrJ) were fed a high-phosphorus diet and, after weaning at 3 weeks of age, were directly fed the high-phosphorus diet. At 4 weeks of age, sodium phytate was continuously administered via the right jugular vein using an osmotic pump (Alzet) at 0.004 mg / body / day, 0.04 mg / body / day, or 0.4 mg / body / day. Arterial calcification was confirmed at 6 weeks of age. Scattered calcification was observed in the aortic arch and the bifurcation of the inferior aorta and femoral artery. Figure 9 shows the aortic arch. (A) Vehicle, (B) 0.4 mg / body / day, and (C) 0.04 mg / body / day. As shown in Figure 9, phytic acid almost completely inhibited vascular calcification stained with Alizarin Red at concentrations of 0.04 mg / body / day or higher.

[0048] (9) Example 9 Bone morphometry using micro-CT Bone morphometry was performed on the proximal tibia of Enpp1 mutant mice (6 weeks old) described above (8). Cortical and cancellous bone were analyzed for bone mass, bone mass / tissue mass, bone mineral density, cancellous bone count, cancellous bone thickness, etc. Bone mass and bone mineral density are shown in Figure 10. (A) represents cortical bone mass, (B) represents cancellous bone mass, (C) represents cortical bone density, and (D) represents cancellous bone density. Phytic acid administration did not affect any of the parameters. Similarly, administration of phytic acid to normal mice did not result in any abnormalities in the various bone morphometric parameters.

[0049] (10) Example 10 Serum iron ion and zinc ion concentrations at effective phytic acid concentrations To confirm the effect of phytic acid on serum iron and zinc concentrations, the following serum samples were analyzed.

[0050] a) Serum samples obtained from mice in Example 7 (1-week continuous intravenous administration) b) Serum samples from Example 8 (2-week continuous intravenous administration) administered with 0.4 mg / body / day sodium phytate and vehicle c) Serum sample from untreated mice fed normal diet at the same time as in Example 8

[0051] Figure 11 shows the results of serum samples administered 0.4 mg / body / day of sodium phytate and vehicle, where (A) shows serum zinc concentration and (B) shows serum iron concentration. Comparison of these samples revealed no effect of phytic acid on serum zinc or iron concentrations.

[0052] The results of this example suggest that phytic acid inhibits ectopic ossification or ectopic calcification, but the main mechanism of action is not the chelating effect of phytic acid, but the inhibition of the expression of bone formation-related factors or calcification-related factors. [Industrial Applicability]

[0053] It can be used to inhibit heterotopic ossification or heterotopic calcification.

Claims

[Claim 1] An agent for inhibiting differentiation of vascular smooth muscle cells into osteoblast-like cells in extraskeletal tissues, characterized by containing phytic acid.

Images