WO2015141874A1 - Pharmaceutical composition for preventing or treating vascular disease containing gemigliptin as active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating vascular disease, the pharmaceutical composition containing, as an active ingredient, gemigliptin which is known as a dipeptidyl peptidase-4 (DPP-4) inhibitor.

Description

Pharmaceutical composition for the prevention or treatment of vascular diseases containing gemigliptin as an active ingredient

The present invention relates to a pharmaceutical composition for the prevention or treatment of vascular disease (gemigliptin) containing gemigliptin known as an inhibitor of dipeptidyl peptidase-4 (DPP-4) as an active ingredient.

Atherosclerosis is a chronic inflammatory disease characterized by the presence of atherosclerotic lesions of the lining of the arteries, leading to hardening and narrowing of the major arteries and ultimately leading to cardiovascular disease ¹ . Abnormal proliferation and migration of vascular smooth muscle cells plays an essential role in the formation of atherosclerotic lesions. In the early stages of arterial vascular wall damage or atherosclerosis, vascular smooth muscle cells in the vascular intima migrate to the endometrium. To proliferate and synthesize extracellular matrix. In addition, the vascular smooth muscle cells in the resting phase are transferred from the G1 phase to the S phase of the cell cycle, and DNA replication is required for proliferation. ² , vascular cell adhesion molecule-1 (VCAM) reacts with various growth factors and cytokines. -1), secreting monocyte chemoattractant protein 1 (MCP-1), which induces inflammation and is accompanied by remodeling to accelerate the progression of atherosclerosis ³ .

Gemigliptin is an inhibitor of DPP-4, a drug for diabetes that was developed based on a mechanism that regulates the amount of insulin secreted by the pancreas by inhibiting the breakdown of glucagon-like peptide-1 (GLP-1). to be. It reported that it was the value of glycated hemoglobin decreased in diabetic patients taking the migeul riptin improve insulin resistance in previous reports ^4, but I study had yet been reported that migeul riptin is involved in the proliferation of vascular smooth muscle cells.

The technical problem to be solved in the present invention is the activity of E2F transcription factor through the inhibition of phosphorylation of Rb (retinoblastoma) of gemigliptin known as DPP-4 inhibitors, which has been disclosed as a possible treatment for diabetes Inhibits the proliferation of vascular smooth muscle cells, inhibits the expression of VCAM-1 and MCP-1 involved in inflammation and remodeling of the vascular lining, and the extracellular air necessary for the proliferation and migration of vascular smooth muscle cells. The present invention is to confirm the inhibition of the activity of matrix metalloproteinase-2 (MMP-2) involved in the degradation and modification of the vagina, and to propose the use as a pharmaceutical composition for the prevention or treatment of vascular diseases.

In order to solve the above technical problem, the present invention provides a pharmaceutical composition for the prevention or treatment of vascular diseases containing gemigliptin (gemigliptin) as an active ingredient.

The vascular disease may be selected from the group consisting of cardiovascular disease, atherosclerosis, carotid artery stenosis, cerebrlvascular stenosis and vascular restenosis.

The gemigliptin may inhibit the proliferation of vascular smooth muscle cells.

Inhibition of proliferation of the vascular smooth muscle cells may be achieved through a pathway that inhibits phosphorylation of Rb.

Inhibition of proliferation of vascular smooth muscle cells can be achieved through a pathway that inhibits the activity of E2F transcription factors.

The gemigliptin may inhibit the migration of vascular smooth muscle cells.

Inhibition of migration of vascular smooth muscle cells can be achieved through a pathway that inhibits expression of VCAM-1.

Inhibition of migration of vascular smooth muscle cells can be achieved through a pathway that inhibits the expression of MCP-1.

Inhibition of migration of vascular smooth muscle cells can be achieved through a pathway that inhibits the activity of MMP-2.

In addition, the present invention provides a method for inhibiting proliferation of vascular smooth muscle cells, characterized in that the treatment of gemigliptin.

The present invention also provides a method of preventing or treating vascular disease by administering gemigliptin.

The method may be targeted to mammals other than humans.

The vascular disease may be selected from the group consisting of cardiovascular disease, atherosclerosis, carotid artery stenosis, cerebrovascular stenosis and vascular restenosis.

According to the present invention, gecamiliptin, known as a DPP-4 inhibitor, inhibits the proliferation of vascular smooth muscle cells by inhibiting the activity of E2F transcription factors through inhibition of phosphorylation of Rb, and VCAM- is involved in inflammation development and modification of vascular inner wall. 1 and MCP-1 inhibits the expression, and acts as a pathway for inhibiting the activity of MMP-2 involved in the decomposition and modification of extracellular matrix required for the proliferation and migration of vascular smooth muscle cells to prevent or treat vascular diseases It may be developed as a suitable composition.

1 is a photograph showing the concentration-dependent decrease in the proliferative activity of vascular smooth muscle cells of rats by the gemigliptin treatment.

Figure 2 is a graph showing the concentration-dependent decrease in the proliferative activity of vascular smooth muscle cells of rats by the gemigliptin treatment.

Figure 3 shows the cell cycle progression of vascular smooth muscle cells of rats treated with gemigliptin treatment, showing that the G1 phase is stagnant and the transition to S and G2 / M phases is reduced.

4 is a result showing that the expression of phosphorylated Rb by gemigliptin treatment is reduced in a concentration-dependent manner.

5 is a result showing that the concentration of E2F by gemigliptin treatment is reduced in a concentration-dependent manner.

Figure 6 is a analysis of the movement of vascular smooth muscle cells through a wound healing assay (wound healing assay), a photograph showing that the movement of cells in the gemigliptin-treated group significantly reduced.

FIG. 7 is a result showing that the activity of MMP-2 increased by tumor necrosis factor-α (TNF-α) is concentration-dependently decreased by gemigliptin treatment.

8 is a result showing that the expression of MCP-1 and VCAM-1 increased by TNF-α is concentration-dependently reduced by gemigliptin treatment.

FIG. 9 schematically shows an experimental procedure for confirming whether gemiliptin inhibits abnormal proliferation of vascular smooth muscle cells in a rat animal model through tissue staining.

FIG. 10 is a result of confirming tissue staining that gemiliptin is administered to the atherosclerosis-induced rats at different concentrations and the degree of neovascularization of vascular smooth muscle cells is reduced.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

The inventors of the present invention, an inhibitor of DPP-4, inhibits the degradation of GLP-1, so that gemigliptin, a diabetic therapeutic agent developed based on a mechanism regulating the amount of insulin secreted from the pancreas, is a cell cycle of vascular smooth muscle cells. reducing the progression of the cycle), reducing the expression of VCAM-1 and MCP-1, and reducing the activity of MMP-2, and the use of gemigliptin as a pharmaceutical composition for the prevention or treatment of vascular diseases I would like to suggest a possibility.

Gemigliptin is a potent, selective, and reversible DPP-4 inhibitor that increases insulin secretion through active GLP-1 and decreases your life and glucose uptake, enhancing your body's ability to regulate blood glucose ⁵ . Recent phase III clinical study showed a sustained and rapid glucose-lowering effect and increase beta-cell sensitivity to the effects of GLP-1 and glucose increases, the stability in toxicity testing has been demonstrated when administered to patients with type 2 diabetes ^6,7. Persistent hyperglycemia is known as a risk factor for microvascular and macrovascular complications, and it is well known that glycemic control reduces the risk of cardiovascular disease ^{8, 9} . However, in addition to hyperglycemia, a variety of risk factors, such as inflammatory cytokines, growth factors, and vascular injuries, change the structure and function of blood vessels and cause narrowing of the blood vessel walls, leading to vascular diseases such as atherosclerosis. It has not been studied whether it is effective in preventing or treating various vascular diseases separately from glycemic control, and such effects have been confirmed by the inventors of the present invention.

Accordingly, the present invention provides a pharmaceutical composition for the prevention or treatment of vascular diseases containing gemigliptin as an active ingredient.

The vascular disease may be selected from the group consisting of cardiovascular disease, atherosclerosis, carotid artery stenosis, cerebrovascular stenosis and vascular restenosis.

When vascular endothelial damage occurs, vascular smooth muscle cells move into the vascular lining by various inflammatory cytokines or growth factors, and then break out of the resting phase and enter a new cell cycle ³ . E2F is a transcription factor that regulates the expression of genes involved in DNA replication, cell proliferation, and the transition from G1 to S phase ¹⁰ . By Rb protein, also known as tumor suppressor will dissociate and in combination with E2F inhibit the transfer ability of E2F, phosphorylated Rb is weaker Rb-E2F inhibitory complex (inhibitory complex) E2F to activate the transcription ability of E2F ^{10 , 11} . In recent years, studies are being actively conducted targeting substances that regulate the cell cycle in vascular cells ¹² . In the study results of the present inventors, the proliferation of vascular smooth muscle cells was reduced by the gemigliptin, and the vascularogenic effects such as the anti-arteriosclerosis effect by demonstrating the mechanism by which the gemigliptin reduced phosphorylated Rb lowers the activity of the E2F transcription factor. The possibility of treating the disease has been presented.

Therefore, the gemigliptin of the present invention can prevent or treat vascular diseases by inhibiting proliferation of vascular smooth muscle cells.

In addition, inhibition of proliferation of the vascular smooth muscle cells may be achieved through a pathway for inhibiting phosphorylation of Rb.

In addition, inhibition of proliferation of the vascular smooth muscle cells may be achieved through a pathway that inhibits the activity of the E2F transcription factor.

Inflammatory responses between inflammatory cells and vascular cells due to endothelial cell damage are also important pathophysiology of early atherosclerosis. Cytokines, such as TNF-α secreted from inflammatory cells, alter the protein expression patterns of vascular smooth muscle cells. Vascular smooth muscle cells activated by TNF-α increase the secretion of VCAM-1 and MCP-1, and consequently accelerate the progression of atherosclerosis with increased proliferation and migration of vascular smooth muscle cells ^{13, 14} . In addition, proliferation and migration of vascular cells requires the degradation and modification of extracellular matrix surrounding the cells, where MMP selectively degrades individual elements of the extracellular matrix ¹⁵ . In particular, MMP-2 plays an important role in the development of atherosclerosis, and previous studies have reported increased expression of MMP-2 by several cytokines including TNF-α in atherosclerotic arteries ¹⁶ . In this study, we confirmed that the increase of MMP-2 activity by TNF-α and the expression of MCP-1 and VCAM-1 were reduced by gemigliptin through cell experiments. It was confirmed that the migration was significantly reduced. Finally, the animal model demonstrated that the degree of neoplastic smooth cells decreased after injection of gemigliptin by concentration.

Therefore, the gemigliptin of the present invention can prevent or treat vascular diseases by inhibiting the migration of vascular smooth muscle cells.

Inhibition of migration of vascular smooth muscle cells can be achieved through a pathway that inhibits expression of VCAM-1.

In addition, the inhibition of migration of the vascular smooth muscle cells may be achieved through a pathway that inhibits the expression of MCP-1.

In addition, the inhibition of migration of vascular smooth muscle cells may be achieved through a pathway that inhibits the activity of MMP-2.

In this study, Gemigliptin, which is used to treat diabetes, reduces the activation of E2F through Rb, and independently inhibits the expression of MMP-2 and the expression of MCP-1 and VCAM-1 independently of blood sugar control. It was confirmed to reduce the proliferation and migration of smooth muscle cells. These findings show that gemigliptin is effective for vascular diseases caused by other causes regardless of diabetes.

The pharmaceutical composition comprising the gemigliptin of the present invention may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and the like, according to a conventional method, for the purpose of each use. It may be formulated and used in various forms, such as injections, and may be administered orally or through various routes including intravenous, intraperitoneal, subcutaneous, rectal, and topical administration.

Such pharmaceutical compositions may further include carriers, excipients or diluents, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, Starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, amorphous cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil Etc. can be mentioned.

In addition, the pharmaceutical composition of the present invention may further include a filler, an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, a preservative, and the like.

In a preferred embodiment, solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which solid preparations comprise at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like are mixed and formulated. In addition, lubricants such as magnesium stearate, talc and the like may also be used in addition to simple excipients.

As a preferred embodiment, oral liquid preparations may be exemplified by suspensions, solvents, emulsions, syrups, and the like, and various excipients, for example, wetting agents, sweeteners, Fragrances, preservatives and the like.

As a preferred embodiment, the preparation for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilizers, suppositories and the like. Non-aqueous solvents and suspending agents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Injectables may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, preservatives, and the like.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, “pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level means the type, severity, activity of the drug, Sensitivity to drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of drugs, and other factors well known in the medical arts. The pharmaceutical compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered as single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, which can be easily determined by those skilled in the art.

In a preferred embodiment, the effective amount of gemigliptin in the pharmaceutical composition of the present invention may vary depending on the age, sex, and weight of the patient, and generally 1 to 5,000 mg, preferably 100 to 3,000 mg per kg body weight. It can be administered daily or every other day or divided into 1 to 3 times a day. However, the dosage may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, etc., and the above dosage does not limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

As used herein, "administration" means providing a patient with any substance by any suitable method, wherein the route of administration of the pharmaceutical composition of the present invention is oral or parenteral via all common routes as long as the target tissue can be reached. Oral administration. In addition, the composition of the present invention may be administered using any device capable of delivering an active ingredient to a target cell.

"Subject" in the present invention is not particularly limited, but includes, for example, humans, monkeys, cattle, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits or guinea pigs. And preferably mammals, and more preferably humans.

In addition, the present invention provides a method for inhibiting proliferation of vascular smooth muscle cells, characterized in that the treatment of gemigliptin.

This method can be used in various animal experiments that meet the in vitro experiments or animal test ethical standards that are not human subjects.

Animals used in the animal experiments are preferably mammals, more preferably monkeys, cows, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, mice, rabbits or guinea pigs. I can lift it.

The present invention also provides a method of preventing or treating vascular disease by administering gemigliptin.

The subject of administration may be a mammal including a human, but may be limited within the scope of a mammal except a human.

The vascular disease may be selected from the group consisting of cardiovascular disease, atherosclerosis, carotid artery stenosis, cerebrovascular stenosis and vascular restenosis.

The method of treatment is achieved by administering gemigliptin to a subject, and the method of administration may be applied according to the method of use of the pharmaceutical composition of the present invention as described above.

Hereinafter, the present invention will be described in more detail with reference to specific examples.

Example

1. Experiment Method

In this study, cell and animal experiments were performed, and all cell experiments were performed using vascular smooth muscle cells (primary RASMC) obtained from the rat aorta. First, the effect of gemigliptin on serum-induced vascular smooth muscle cell proliferation was observed using flow cytometry (FACs). Western blot and promoter activity were used to measure Rb expression and E2F. Activity was measured. In addition, the expression patterns of VCAM1 and MCP1, which are inflammation-related factors induced by TNF-, were examined, and MMP-2 activity and cell migration were also examined. Finally, in animal experiments, induction of arteriosclerosis by binding the common carotid artery of rats was induced. It was confirmed by staining (H & E stain).

2. Experimental Results

2.1. Inhibition of Vascular Smooth Muscle Cell Proliferation by Gemigliptin Treatment

The number and proliferative activity of the cells in the gemigliptin-treated group decreased in a concentration-dependent manner compared to the non-groups (Figs. 1 and 2). It was confirmed that the transition to the / M stage is reduced (Fig. 3).

In addition, concentration-dependent phosphorylated Rb after gemigliptin treatment decreased (FIG. 4) and E2F promoter activity also decreased (FIG. 5).

2.2. Inhibition of Vascular Smooth Muscle Cells by Gemigliptin Treatment

The movement of cells was analyzed through wound healing analysis, and it was confirmed that the movement of cells was significantly reduced in the sample treated with gemigliptin (FIG. 6). As a result of examining the effect of Gemigliptin on TNF-α-induced MMP-2 activity and MCP-1 and VCAM-1 expression, Gemigliptin dose-dependently decreased MMP-2 activity (FIG. 7). In addition, mRNA expression of MCP-1 and VCAM-1 was also reduced (FIG. 8).

2.3. Effect of Gemigliptin Administration in Animal Models

Tissue staining confirmed that gemigliptin inhibited abnormal proliferation of vascular smooth muscle cells in animal models. The animal model induced atherosclerosis through the CCA ligation of rats. Gemigliptin was fed to the diet by concentration (0, 0.04, 0.09, 0.27%) and anesthetized after 2 weeks. (FIG. 9). When the neovascularization of vascular smooth muscle cells in the group administered with gemigliptin was compared after tissue staining, it was confirmed that the degree of neoplastic cells in the group administered with gemigliptin was lowered (FIG. 10).

references

The references listed below are materials for more specifically describing the present invention, and the entire contents of the listed references are incorporated as a whole into the specification of the present invention.

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Claims (12)

1. A pharmaceutical composition for preventing or treating vascular disease containing gemigliptin as an active ingredient.
2. The group of claim 1, wherein the vascular disease comprises cardiovascular disease, atherosclerosis, carotid artery stenosis, cerebrlvascular stenosis, and vascular restenosis. A pharmaceutical composition, characterized in that selected from.
3. The pharmaceutical composition of claim 1, wherein the gemigliptin inhibits proliferation of vascular smooth muscle cells.
4. The pharmaceutical composition of claim 3, wherein the inhibition of proliferation of the vascular smooth muscle cells is achieved through a pathway for inhibiting phosphorylation of retinoblastoma (Rb).
5. 4. The pharmaceutical composition of claim 3, wherein the inhibition of proliferation of vascular smooth muscle cells is achieved through a pathway that inhibits the activity of an E2F transcription factor.
6. The pharmaceutical composition of claim 1, wherein the gemigliptin inhibits the movement of vascular smooth muscle cells.
7. 7. The pharmaceutical composition of claim 6, wherein the inhibition of vascular smooth muscle cell migration is through a pathway that inhibits expression of vascular cell adhesion molecule-1 (VCAM-1).
8. The pharmaceutical composition according to claim 6, wherein the inhibition of vascular smooth muscle cell migration is through a path of inhibiting the expression of monocyte chemoattractant protein-1 (MCP-1).
9. 7. The pharmaceutical composition of claim 6, wherein the inhibition of vascular smooth muscle cell migration is achieved through a pathway that inhibits the activity of matrix metalloproteinase-2 (MMP-2).
10. A method of inhibiting proliferation of vascular smooth muscle cells, characterized by treating gemigliptin.
11. A method of preventing or treating vascular disease by administering gemigliptin to a mammal other than a human.
12. The group of claim 11, wherein the vascular disease comprises cardiovascular disease, atherosclerosis, carotid artery stenosis, cerebrlvascular stenosis, and vascular restenosis. Selected from

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