KR100922507B1 - Novel Compound and Pharmaceutical Composition Comprising the Same for the Prevention and Treatment of Vascular Smooth Muscle Cell Hyper-Proliferative Disease - Google Patents

Abstract

The present invention relates to a novel compound, a composition for inhibiting vascular smooth muscle cell proliferation comprising the compound as an active ingredient, and a pharmaceutical composition for preventing or treating vascular smooth muscle cell abnormal proliferative disease. Since the active ingredient compound of the present invention has vascular smooth muscle cell proliferation inhibitory activity, it can be very useful for the treatment or prevention of abnormal proliferative diseases of vascular smooth muscle cell.

Description

Novel Compound and Pharmaceutical Composition Comprising the Same for the Prevention and Treatment of Vascular Smooth Muscle Cell Hyper-Proliferative Disease

The present invention relates to a novel compound, a composition for inhibiting vascular smooth muscle cell proliferation comprising the compound as an active ingredient, and a pharmaceutical composition for preventing or treating vascular smooth muscle cell abnormal proliferative disease containing the compound as an active ingredient.

Cardiovascular diseases, including heart failure, hypertensive heart disease, arrhythmia, congenital heart disease, myocardial infarction and angina, and vascular diseases such as arteriosclerosis, stroke, and peripheral vascular disease, are widely referred to as ischemic cardiovascular disease. . These diseases occur in various age groups and can lead to death with serious sequelae if not treated in a timely manner. Due to westernized dietary and lifestyle changes, the incidence and mortality rate of cardiovascular diseases continues to increase rapidly. However, effective therapeutics for treating such diseases have not been developed.

Current treatments for coronary artery disease include pharmacotherapy, gene therapy, scenic coronary angioplasty and stenting (PTCA), and surgical surgical coronary artery bypass graft (CABG). Revascularization therapy is underway. However, even after reperfusion therapy, the drug should be continued and there is a possibility of recurrence of coronary artery disease.

In particular, percutaneous coronary angioplasty (PTCA) has been widely used since 1977 as a treatment for patients with coronary atherosclerosis (Gurentzig, Lancet, 4263, 1978). There is a problem that restenosis occurs in about 40% of patients after -6 months (Ryan et al., J. Am. Coll. Cardiol., 22. 2033-2054, 1993). The mechanism of the development of restenosis after PTCA is to promote the migration and proliferation of vascular smooth muscle cells by producing and secreting various cytokines from vascular endothelial cells, activated platelets or macrophages damaged by a balloon. It is known that the production and secretion of the extracellular matrix (ECM) is promoted, leading to thickening of the vascular lining and developing into restenosis (Godfried et al, Am. Heart J., 129, 203-210, 1995).

Normal vascular smooth muscle cells do not proliferate, but endothelial cell membranes are damaged through stents, resulting in multiple stages of signal transduction resulting in the division, migration and proliferation of vascular smooth muscle cells. The mechanisms of vascular smooth muscle cell proliferation include the removal of proliferation inhibitory factor due to normal endothelial cell damage and the activation of proliferation-inducing factors, the transmission of proliferative signals through receptors on the surface of vascular smooth muscle cells, and the proliferation of vascular smooth muscle cells into the nucleus. And a change in the cell cycle caused by the trigger signal. Normal endothelial cells secrete substances that inhibit the proliferation of vascular smooth muscle cells, and when the endothelial cells are damaged, their secretion is suppressed, and platelet derived growth factor, which is secreted from activated platelets, is contained in plasma. It is known that the proliferation of vascular smooth muscle cells is caused by various cytokines.

To date, attempts have been made to prevent restenosis using antiplatelet agents, anticoagulants, cholesterol biosynthesis inhibitors, and antiallergic agents (Lefkovits et al., Progr. Cardiovas. Dis., 40, 141-158, 1997). There is research on the prevention of restenosis of natural-derived components such as fish oil, vitamin C and vitamin E. However, these drugs have not yet obtained sufficient results in clinical practice.

In addition, Taxol (Herdeg et al., Zeischrift fur Kardiologie, 89, 390-397, 1999), heparin, EPA, estrogen, etc. are under research and development as inhibitors of vascular restenosis, but this also does not have a clear effect. However, Korean Patent Registration No. 478671 discloses a pharmaceutical composition for preventing and treating vascular restenosis including clotrimazole as an active ingredient, and Korean Patent Registration No. 516026 discloses 3'-deoxyadenosine. A preventive and therapeutic composition for vascular restenosis is disclosed as an active ingredient, and Korean Unexamined Patent Publication No. 2001-110793 discloses an agent for preventing and treating vascular stenosis using an active compound having Rho kinase inhibitory activity. . In addition, Korean Patent Laid-Open Publication No. 2003-46314 discloses the prevention and treatment of vascular dysplasia such as transplanted vascular disease, restenosis, in-stent restenosis, and pulmonary hypertension with antithrombin as an active ingredient. A composition for a treatment is disclosed, and Korean Patent Laid-Open Publication No. 2005-23249 discloses prevention of vascular diseases such as stenosis and / or reclosure after percutaneous coronary angioplasty using an vascular stent using a retinoid and a retinoid action modifier. And / or a pharmaceutical composition for the treatment is disclosed, Korean Patent Publication No. 2005-43183 discloses a pharmaceutical composition for preventing and treating vascular restenosis, including curcumin (curcumin). However, in addition to preventing vascular restenosis, these vascular restenosis inhibitors have various side effects such as inhibiting wound regeneration, vascular injury, hepatotoxicity, kidney damage, and increased bleeding by platelet aggregation. Although research is being actively conducted to develop a substance capable of inhibiting vascular restenosis derived from the various proven natural products, no research results have been reported. Therefore, there is a constant need to develop materials that can safely and effectively inhibit vascular restenosis.

In addition, atherosclerosis is a disease in which the elasticity of the artery wall is lost and the wall is hardened and hardened. The arteriosclerosis exhibits various symptoms depending on the organs involved, but is most commonly caused by ischemia due to a lack of blood supply and loss of elasticity. Because of the symptoms of rupture hemorrhage. In particular, if the coronary artery causes arteriosclerosis and blood flow is reduced, angina pectoris is felt during exercise, and if it is further reduced, it becomes unstable angina, which causes pain even at rest, and if it is very blocked, myocardial infarction This can put your life at risk.

In the meantime, researches on treating such arteriosclerosis have been conducted. In the 1970s, the tube was inserted into the heart, angiography was discovered, and the lesion was found. The idea of a method of physically widening the balloon was introduced. Recently, drug-coated stents have been used to increase the success rate of the procedure, and the success rate of the procedure in a wider range of situations is increasing, leading to the treatment of heart disease. Although the surgical technique for atherosclerosis has made such a lot of progress, the academic achievements to prevent and control arteriosclerosis itself still have much to be solved, and studies on the causes and treatment of atherosclerosis, Research and application of effective antithrombotic and anticoagulant agents to inhibit thrombus formation are needed.

Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

The present inventors have made diligent efforts to develop substances that can effectively treat and prevent diseases caused by excessive proliferation of vascular smooth muscle cells. As a result, the compounds of the present invention have a proliferation inhibitory activity of vascular smooth muscle cells (vsmc). By experimentally confirming the present invention, the present invention was completed.

It is therefore an object of the present invention to provide novel compounds.

Another object of the present invention to provide a pharmaceutical composition for the prevention and treatment of vascular smooth muscle cell abnormal proliferative disease containing the compound of the present invention as an active ingredient.

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The objects and advantages of the invention will become apparent from the following detailed description, claims and drawings.

According to one aspect of the present invention, the present invention provides novel compounds represented by the following general formula (I) or (II).

[Formula I] [Formula II]

However, in formula (I), R ₁ is C ₁ -C ₄ linear or branched alkyl.

According to another aspect of the present invention, the present invention provides a pharmaceutical composition comprising (a) a pharmaceutically effective amount of any one of the compounds represented by the following formulas (I), (II) and (III); And (b) provides a pharmaceutical composition for the prevention or treatment of vascular smooth muscle cell hyper-proliferative disorder comprising a pharmaceutically acceptable carrier.

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[Formula I] [Formula II]

[Formula III]

However, in formula (I), R ₁ is C ₁ -C ₄ linear or branched alkyl.

The present inventors have made diligent efforts to develop substances that can effectively suppress diseases caused by excessive proliferation of vascular smooth muscle cells (VSMCs), and thus the compounds of the present invention have excellent vascular smooth muscle cell proliferation inhibitory activity. The present invention was completed by experimentally confirming.

The compound of the present invention is a compound represented by formula (I), or formula (II) or formula (III). In the general formula (I) defining compounds of the present invention, the term "C ₁ -C ₄ straight or branched chain alkyl" refers to a straight or branched chain saturated hydrocarbon having 1 to 4 carbon atoms, for example methyl, ethyl , n-propyl, isopropyl, isobutyl, n-butyl and t-butyl.

Preferred compounds of the formula (I) of the present invention are compounds represented by the following formula (Ia), (Ib), or (Ic).

Formula Ia Formula Ib Formula Ic

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The compound of the present invention is a novel compound synthesized by the inventors for the first time. Specific examples of the compound represented by the formula (I) of the present invention can be synthesized by the following reaction scheme.

In the above scheme, a is boronic acid / dichloromethane, b is boron tribromide / dichloromethane.

Compound 1 (3-Hydroxy-4,5-dimethoxy-benzoic acid methyl ester) used as a starting material in the scheme can be easily prepared by methylation in gallate. Compound 2 in the above scheme can be prepared by reacting compound 1 with boronic acid in dichloromethane solvent. The boronic acid is a compound in which C ₁ -C ₃ straight chain, C ₁ -C ₄ straight chain or branched alkyl is substituted. The reaction can be carried out at room temperature for 4-6 hours, and the reactor must be designed to allow oxygen to flow. The borotribromide can be used in an amount of 1.2-1.5 equivalents based on compound 2.

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Since the compounds of the present invention have a proliferation inhibitory activity of vascular smooth muscle cells, it can be very useful in various fields where excessive proliferation of vascular smooth muscle cells needs to be suppressed.

In addition, the compounds of the present invention can be used for the prevention or treatment of vascular smooth muscle cell hyper-proliferative disorder.

The compound of the present invention has a proliferation inhibitory activity of vascular smooth muscle cells. According to a specific embodiment of the present invention, the compound of the present invention exhibits the effect of inhibiting the proliferation of aortic smooth muscle cells of white rats induced by platelet-derived growth factor (PDGF-BB).

According to another specific embodiment of the present invention, the compound of the present invention inhibits the incorporation of [ ³ H] thymidine into DNA upon proliferation of aortic smooth muscle cells of white rats induced by PDGF-BB.

In the present specification, the term "smooth muscle" refers to organs such as tubular organs, bladder, abdominal cavity, uterus, reproductive tract, gastrointestinal tract, respiratory tract, vasculature, skin and ciliary muscle, eye iris, etc. non-striated muscle found in the walls of organs. The smooth muscle is spindle shaped and is capable of contraction and relaxation.

The cells constituting the smooth muscle "smooth muscle cell" is a monocyte, characterized in that it is arranged in the form of a sheet (sheet) or bundle (bundle) is connected by a gap junction (gap junction).

The term "vascular smooth muscle cell" (VSMC) in the context of the present invention refers to the cells that make up the smooth muscle of the vessel wall.

The term "VSMC hyper-proliferative disorder" in the context of the present invention means a disease or condition resulting from excessive proliferation of vascular smooth muscle cells.

In the present invention, "vascular smooth muscle cell abnormal proliferative disease" includes various diseases. For example, vascular restenosis, vascular stenosis, arteriosclerosis, atherosclerosis, which are directly caused by abnormal proliferation of vascular smooth muscle cells, as well as cardiovascular diseases secondary to stenosis or atherosclerosis, heart failure, myocardium Infarctions, angina pectoris, arrhythmia, hypertensive heart disease, congenital heart disease, stroke, peripheral vascular stenosis, and the like.

Preferably, the vascular smooth muscle cell aberrant proliferative disease treated or prevented by the pharmaceutical composition of the present invention is atherosclerosis, atherosclerosis, vascular restenosis or vascular stenosis.

Atherosclerosis is a disease in which fatty substances are deposited or fibrosis in the inner layers of arteries. On the other hand, restenosis of blood vessels is a disease in which the vascular pathway is narrowed after traumatization. Vascular restenosis occurring after atherosclerosis and stent implantation is known to be due to the proliferation, migration and secretion of extracellular matrix of vascular smooth muscle cells (Circulation, 1997, 95, 1998-2002; Clin. Invest. 1997, 99, 2814-2816; Cardiovasc. Res. 2002, 54, 499-502). In order to prevent the progression of arteriosclerosis and vascular restenosis, studies on drugs that inhibit the proliferation of vascular smooth muscle cells have been widely conducted. Currently, several drugs have been used for the treatment of patients (J. Am. Coll. Cardiol., 2002, 39, 183-193). Therefore, it can be seen that the compounds of the present invention which effectively inhibit the proliferation of vascular smooth muscle cells are very effective for the treatment of vascular smooth muscle cell abnormal proliferative diseases.

The pharmaceutical composition for preventing or treating vascular smooth muscle cell abnormality proliferative disease of the present invention comprises an active ingredient and a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and when administered parenterally, may be administered by intravenous infusion, subcutaneous infusion, intramuscular injection, intraperitoneal injection, transdermal administration, or the like. It is preferable that the route of administration of the pharmaceutical composition of the present invention is determined according to the type of the disease to be applied.

The concentration of the compound which is the active ingredient included in the composition of the present invention can be determined in consideration of the purpose of treatment, the condition of the patient, the period of time, etc., and is not limited to a specific range of concentration. However, preferably the active ingredient compound of the present invention may be added at a concentration of 0.5-3 μM. If it is contained in a concentration of less than 0.5 μM it is difficult to expect the desired effect, if it is contained in a concentration of more than 3 μM it is difficult to expect an increased effect in proportion to the increase in concentration and also increase the manufacturing cost of the product according to the invention There are disadvantages.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or an aqueous medium, or may be in the form of extracts, powders, granules, tablets or capsules, and may further include a dispersant or stabilizer.

The present invention provides a novel compound, a composition for inhibiting vascular smooth muscle cell proliferation comprising the compound as an active ingredient, and a pharmaceutical composition for preventing and treating vascular smooth muscle cell abnormal proliferative disease comprising the compound as an active ingredient.

Since the novel compounds of the present invention have excellent vascular smooth muscle cell proliferation inhibitory activity, arteriosclerosis, atherosclerosis, vascular restenosis, vascular stenosis and various cardiovascular diseases secondary to the vascular smooth muscle cell abnormality proliferative diseases It can be very useful for the prevention or treatment of

Figures 1a-1e show the aortic smooth muscle of white rats at concentrations of 0.5 μM, 1 μM, 3 μM of JY0691, JY0692, JY0693, JY0694, JY0695, which are represented by the compounds of the present invention (Ia, Ib, Ic, II, III) It is a graph which shows the proliferation inhibitory effect of this cell after processing to a cell.

Figures 2a-2e shows the aortic smooth muscle of white rats at concentrations of 0.5 μM, 1 μM, 3 μM of JY0691, JY0692, JY0693, JY0694, JY0695 represented by the compounds of the present invention (Ia, Ib, Ic, II, III) This graph shows the DNA synthesis inhibitory effect of the cells after treatment.

Figure 3a-3e is a compound of the present invention represented by the formulas Ia, Ib, Ic, II, III JY0691, JY0692, JY0693, JY0694, JY0695 in a concentration of 0.5 μM, 1 μM, 3 μM white rat aortic smooth muscle cells A graph showing the effect on the cell proliferation cycle of these cells after treatment with.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example  1: 3- (4- propylphenoxy ) -4,5- dimethoxy - benzoic acid methyl ester Synthesis of

In the scheme, a is 4-propyl boronic acid (4-propyl boronic acid) / dichloromethane (CH ₂ Cl ₂ ).

3-Hydroxy-4,5-dimethoxy-benzoic acid methyl ester (compound of formula 1) (1.0 eq), 4-propylboronic acid (4-propylboronic acid) (1.0 eq), cooperacetate (0.1 eq) and triethylamine (5.0 eq) were added and dissolved in dichloromethane. At this time, the device is allowed to get oxygen into the flask and a proper amount of 4 mo molecular sieve is added. The reaction was filtered after 2.5 hours at room temperature. The filtrate was concentrated under reduced pressure and then separated by column chromatography. The resulting compound was yellow oily form, NMR analysis of the compound was as follows. Based on the NMR analysis, the synthesized compound was identified to be a compound of 3- (4-Propylphenoxy) -4,5-dimethoxy-benzoic acid methyl ester.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.39 (s, 1H, Ar- H ), 7.29 (s, 1H, Ar- H ), 7.12 (d, 2H, J = 8.4 Hz, Ar- H ), 6.88 (d, 2H, J = 8.4 Hz, Ar- H ), 3.93 (s, 3H, -O CH ₃ ), 3.91 (s, 3H, -O CH ₃ ), 3.85 (s, 3H, -COO CH ₃ ) , 2.58 - 2.48 (m, 2H , Ar- CH 2 CH 2 CH 3), 1.68 - 1.53 (m, 2H, Ar-CH 2 CH 2 CH 3), 0.96 - 0.85 (m, 3H, Ar-CH 2 CH ₂ CH ₃ ); IR (NaCl) 2931, 1714, 1220, 1091 cm ^-1

Example 2 Synthesis of 3- (4-Butylphenoxy) -4,5-dimethoxy-benzoic acid methyl ester

In the scheme, a is 4-butyl boronic acid (4-butyl boronic acid) / dichloromethane (CH ₂ Cl ₂ ).

3-Hydroxy-4,5-dimethoxy-benzoic acid methyl ester (compound of formula 1) (1.0 eq), 4-butylboronic Acid (4-butyl boronicacid) (1.0 eq), cupperacetate (0.1 eq) and triethylamine (5.0 eq) were added and dissolved in dichloromethane. At this time, oxygen was introduced into the flask and a 4 Å molecular sieve was added. The reaction was filtered after 2.5 hours at room temperature. The filtrate was concentrated under reduced pressure and then separated by column chromatography. The resulting compound was yellow oily form, NMR analysis of the compound was as follows. Based on NMR analysis, the synthesized compound was identified to be 3- (4-Butylphenoxy) -4,5-dimethoxy-benzoic acid methyl ester.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.39 (s, 1H, Ar- H ), 7.29 (s, 1H, Ar- H ), 7.12 (d, 2H, J = 8.4 Hz, Ar- H ), 6.68 (d, 2H, J = 8.4 Hz, Ar- H ), 3.93 (s, 3H, -OC H ₃ ), 3.91 (s, 3H, -OC H ₃ ), 3.85 (s, 3H, -COOC H ₃ ) , 2.60-2.50 (m, 2H, Ar-C H ₂ CH ₂ CH ₂ CH ₃ ), 1.60-1.46 (m, 2H, Ar-CH ₂ C H ₂ CH ₂ CH ₃ ), 1.41-1.31 (m, 2H , Ar—CH ₂ CH ₂ C H ₂ CH ₃ ), 0.94-0.85 (m, 3H, Ar—CH ₂ CH ₂ CH ₂ C H ₃ ); IR (NaCl) 2944, 1720, 1220, 1099 cm ^-1

Example 3 Synthesis of 3- (4-tert-Butylphenoxy) -4,5-dimethoxy-benzoic acid methyl ester

In the scheme, a is 4-tert-butyl boronic acid / dichloromethane (CH ₂ Cl ₂ ).

3-Hydroxy-4,5-dimethoxy-benzoic acid methyl ester (compound of formula 1) (1.0 eq), 4-tertbutylboro Nickel acid (4-tert-butyl boronicacid) (1.0 eq), cooperacetate (0.1 eq) and triethylamine (5.0 eq) were added and dissolved in dichloromethane. At this time, oxygen was introduced into the flask and a 4 Å molecular sieve was added. The reaction was filtered after 2.5 hours at room temperature. The filtrate was concentrated under reduced pressure and then separated by column chromatography. The resulting compound was yellow oily form, NMR analysis of the compound was as follows. Based on the NMR analysis, the synthesized compound was identified to be a compound of 3- (4-tert-butylphenoxy) -4,5-dimethoxy-benzoic acid methyl ester.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.41 (s, 1H, Ar- H ), 7.33 (d, 2H, J = 8.8 Hz, Ar- H ), 7.27 (s, 1H, Ar- H ), 6.90 (d, 2H, J = 8.8 Hz, Ar- H ), 3.95 (s, 3H, -COOC H ₃ ), 3.93 (s, 3H, -OC H ₃ ), 3.89 (s, 3H, -OC H ₃ ) , 1.31 (s, 9H, Ar—C (C H ₃ ) ₃ ); IR (NaCl) 1722, 1425, 1222, 1099 cm ^-1

Example  4: 3- (4- Propylphenoxy ) -4,5- dihydroxy - benzoic acid methyl ester Synthesis of

In the scheme b is boron tribromide / dichloromethane (CH ₂ Cl ₂ ).

Compound 3- (4-propylphenoxy) -4,5-dimethoxy-benzoic acid methyl ester synthesized in Example 1 above (3- (4-propylphenoxy) -4,5-dimethoxy-benzoic acid methyl ester (1.0 eq) was dissolved in dichloromethane under a stream of nitrogen. BBr ₃ was added dropwise at -78 ° C to a dichloromethane solution. After stirring for 30 minutes, the mixture was reacted at room temperature for 2 hours. After the reaction was quenched with methanol (methanol) and stirred for 20 minutes. Extraction was performed three times with ethyl acetate and concentrated under reduced pressure to obtain a product by column chromatography. The NMR analysis of the compound was as follows. Based on the NMR analysis results, it was confirmed that the synthesized compound is a compound of 3- (4-propylphenoxy) -4,5-dihydroxy-benzoic acid methyl ester.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.42 (s, 1H, Ar- H ), 7.16 (d, 2H, J = 8.4 Hz, Ar- H ), 7.16 (s, 1H, Ar- H ), 6.94 (d, 2H, J = 8.4 Hz, Ar- H ), 5.99 (s, 1H, -O H ), 5.56 (s, 1H, -O H ), 3.81 (s, 3H, -OC H ₃ ), 2.57 (t, 2H, J = 7.6 Hz, Ar-C H ₂ CH ₂ CH ₃ ), 1.70-1.57 (m, 2H, Ar -CH ₂ C H ₂ CH ₃ ), 0.95 (t, 3H, J = 7.3 Hz , Ar—CH ₂ CH ₂ C H ₃ ); IR (KBr) 3340, 2929, 1689, 1253 cm ^-1

Example  5: 3- (4- Butylphenoxy ) -4,5- dihydroxy - benzoic acid methyl ester Synthesis of

In the scheme b is boron tribromide / dichloromethane (CH ₂ Cl ₂ ).

Compound 3- (4-Butylphenoxy) -4,5-dimethoxy-benzoic acid methyl ester (1.0 eq) synthesized in Example 2 was dissolved in dichloromethane under a nitrogen stream. BBr ₃ was added dropwise at -78 ° C to a dichloromethane solution. After stirring for 30 minutes, the mixture was reacted at room temperature for 2 hours. After completion of the reaction, the mixture was quenched with methanol and stirred for 20 minutes. After extraction three times with ethyl acetate and concentrated under reduced pressure to obtain a product by column chromatography (column chromatography) to obtain the product, the NMR analysis of the compound was as follows. Based on NMR analysis, the synthesized compound was identified to be a compound of 3- (4-Butylphenoxy) -4,5-dihydroxy-benzoic acid methyl ester.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.42 (s, 1H, Ar- H ), 7.16 (d, 2H, J = 8.4 Hz, Ar- H ), 7.16 (s, 1H, Ar- H ), 6.94 (d, 2H, J = 8.4 Hz, Ar- H ), 5.95 (s, 1H, -O H ), 5.49 (s, 1H, -O H ), 3.82 (s, 3H, -OC H ₃ ), 2.60 (t, 2H, J = 7.7 Hz, Ar-C H ₂ CH ₂ CH ₂ CH ₃ ), 1.65-1.55 (m, 2H, Ar-CH ₂ C H ₂ CH ₂ CH ₃ ), 1.42-1.25 (m, 2H, Ar-CH ₂ CH ₂ C H ₂ CH ₃ ), 0.85 (t, 3H, J = 8.0 Hz, Ar-CH ₂ CH ₂ CH ₂ C H ₃ )

Example  6: 3- (4- tert - Butylphenoxy ) -4,5- dihydroxy - benzoic acid methyl  Synthesis of Ester

In the above scheme b is boron tribromide / dichloromethane (CH ₂ Cl ₂ ).

Compound 3- (4-tert-Butylphenoxy) -4,5-dimethoxy-benzoic acid methyl ester (1.0 eq) synthesized in Example 3 was dissolved in dichloromethane under a nitrogen stream. BBr ₃ was added dropwise at -78 ° C to a dichloromethane solution. After stirring for 30 minutes, the mixture was reacted at room temperature for 2 hours. After completion of the reaction, the mixture was quenched with methanol and stirred for 20 minutes. Extraction was performed three times with ethyl acetate, and the residue was concentrated under reduced pressure to obtain a product by column chromatography. The NMR analysis of the compound was as follows. Based on NMR analysis, the synthesized compound was identified to be 3- (4-tert-Butylphenoxy) -4,5-dihydroxy-benzoic acid methyl ester.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.42 (s, 1H, Ar- H ), 7.37 (d, 2H, J = 8.6 Hz, Ar- H ), 7.19 (s, 1H, Ar- H ), 6.96 (d, 2H, J = 8.6 Hz, Ar- H ), 5.96 (s, 1H, -O H ), 5.52 (s, 1H, -O H ), 3.83 (s, 3H, -OC H ₃ ), 1.33 (s, 9H, Ar-C (C H ₃ ) ₃ ); IR (KBr) 3313, 1700, 1444, 1103 cm ^-1

Example 7 Synthesis of 5-Allyl-3- (4-allylphenoxy) -1,2-phenylene diacetate

In the scheme c is acetic anhydride (Ac ₂ O), triethylamine (triethyl amine, Et ₃ N), 4-dimethylaminopyridine (4-dimethylaminopyridine, 4-DMAP) / dichloromethane (CH ₂ Cl ₂ ).

To a dichloromethane solution of Obovatol (5-allyl-3- (4-allylphenoxy) benzene-1,2-diol) (1.0 eq), 4-dimethylaminopyridine (0.2 eq), triethylamine ( 5.0 eq) and acetic anhydride (2.2 eq) were added and reacted at room temperature. After stirring for 20 minutes, the reaction was terminated and neutralized with sodium bicarbonate (NaHCO ₃ ). Then extracted twice with ethyl acetate, concentrated under reduced pressure and separated by column chromatography. The resulting compound was yellow oily form, NMR analysis of the compound was as follows. Based on the NMR analysis results, the synthesized compound was identified as a compound of 5-allyl-3- (4-allylphenoxy) -1,2-phenylene diacetate.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.14 (d, 2H, J = 8.4 Hz, Ar-H), 6.95 (d, 2H, J = 8.4 Hz, Ar-H), 6.76 (s, 1H, Ar -H), 6.65 (s, 1H, Ar-H), 6.03-5.82 (m, 2H, Ar-CH ₂ CH CH ₂ ), 5.06 (d, 4H, J = 15.2 Hz, Ar- CH ₂ CHCH ₂ ) , 3.38 (d, 2H, J = 6.6 Hz, Ar-CH ₂ CH CH ₂ ), 3.29 (d, 2H, J = 6.6 Hz, Ar-CH ₂ CH CH ₂ ), 2.29 (s, 3H, -OCOCH ₃ ), 2.26 (s, 3H, -OCOCH ₃ )

Example  8: 5- Allyl -3- (4- allylphenoxy ) -1,2- phenylene diacetate Synthesis of

In the above scheme d is palladium-carbon (Pd-C), hydrogen (H ₂ ) / ethanol (Ethanol).

Obovatol (5-allyl-3- (4-allylphenoxy) benzene-1,2-diol) Palladium-carbon (Pd-C) was added to an ethanol solution of (1.0 eq), and reduced pressure (H ₂ ) was substituted. After reacting for 24 hours under hydrogen stream, the mixture was filtered using celite. The filtrate was concentrated under reduced pressure, and then separated by column chromatography. The resulting compound was oily, and the NMR analysis of the compound was as follows. Based on the NMR analysis results, it was confirmed that the synthesized compound was a compound of 5-propyl-3- (4-propylphenoxy) benzene-1,2-diol.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.14 (d, 2H, J = 8.4 Hz, Ar-H), 6.93 (d, 2H, J = 8.4 Hz, Ar-H), 6.56 (s, 1H, Ar -H), 6.28 (s, 1H, Ar-H), 2.57 (t, 2H, J = 7.6 Hz, Ar- CH ₂ CH ₂ CH ₃ ), 2.41 (t, 2H, J = 7.6 Hz, Ar- CH ₂ CH ₂ CH ₃ ), 1.70-1.42 (m, 4H, Ar-CH ₂ CH ₂ CH ₃ ), 0.95 (t, 3H, J = 7.3 Hz, Ar-CH ₂ CH ₂ CH ₃ ), 0.88 (t, 3H, J = 7.3 Hz, Ar-CH ₂ CH ₂ CH ₃ )

Example  9: aorta Smooth muscle cell  Effect on growth inhibition JY0691 , JY0692 , JY0693, JY0694 , JY0695 Effect

Smooth muscle cell proliferation inhibitory effect on the compound synthesized through the reaction process of Example 1-8 was measured.

The compounds synthesized in Examples 4, 5, 6, 7 and 8 were named JY0691, JY0692, JY0693, JY0694, and JY0695, respectively. To determine whether these compounds inhibit the growth of aortic smooth muscle cells in white rats, 4.0 × 10 ⁴ cells / ml of aortic smooth muscle cells were transferred to 12 well plates and 0.5% (V / V) fetal calf Incubated for 24 hours in DMEM medium containing serum. Subsequently, the medium was treated with compounds JY0691, JY0692, JY0693, JY0694, JY0695 at concentrations of 0.5, 1, and 3 μM, and then further incubated for 24 hours, followed by platelet-dereived growth factor (PDGF). -BB) and after 24 hours with trypsin to separate the cells from the bottom and count the number of isolated cells using a hemocytometer.

1A to 1E show the results of the proliferation inhibitory effect of the cells of the present invention after treatment with the compounds JY0691, JY0692, JY0693, JY0694, JY0695 at concentrations of 0.5 μM, 1 μM, and 3 μM of rat aortic smooth muscle cells. The graph shown. From the results of FIGS. 1A-1E, it was confirmed that the compounds JY0691, JY0692, JY0693, JY0694, and JY0695 of the Examples of the present invention effectively inhibit aberrant proliferation of aortic smooth muscle cells by platelet-derived growth factors at concentrations of 0.5-3 μM. .

Example  10: JY0691 , JY0692 , JY0693 , JY0694 , JY0695 On DNA Synthesis of Aortic Smooth Muscle Cells in Rats

In Example 10, it was confirmed that the compounds JY0691, JY0692, JY0693, JY0694, JY0695 inhibit the proliferation of aortic smooth muscle cells in white rats. Effect of Compounds JY0691, JY0692, JY0693, JY0694, JY0695 on DNA Biosynthesis during Cell Proliferation to Determine What Mechanism Prohibits Proliferation Inhibition of Aortic Smooth Muscle Cells of Compounds JY0691, JY0692, JY0693, JY0694, JY0695 Learned.

2.0 x 10 ⁴ cells / ml of aortic smooth muscle cells were transferred to 24-well plates and incubated for 24 hours in DMEM medium containing 0.5% (V / V) fetal calf serum. Subsequently, compounds JY0691, JY0692, JY0693, JY0694, and JY0695 were treated at concentrations of 0.5, 1, and 3 μM, respectively, incubated for 24 hours, and then platelet-dereived growth factor (PDGF-BB). Treated. After 20 hours, 1 μL of [ ³ H] thymidine was treated. After 4 hours incubation, washed twice with PBS and treated with TCA reagent for 30 minutes. After removing the TCA reagent, it was washed twice with ethanol / ether (1: 1 v / v). 800 μL of 1 N NaOH was dispensed into each well to completely lyse the cells, and then mixed with a 4 ml scintillation cocktail to measure radioactivity using a liquid scintillation counter. It was.

Figures 2a-2e is a graph showing the effect of inhibiting DNA synthesis of the cells after treatment of rat aortic smooth muscle cells in concentrations of 0.5 μM, 1 μM, 3 μM of compounds JY0691, JY0692, JY0693, JY0694, JY0695. From the results shown in Figures 2a to 2e, it can be seen that the compounds of the present invention JY0691, JY0692, JY0693, JY0694, JY0695 very effectively inhibit the DNA biosynthesis of aortic smooth muscle cells in the concentration range of 0.5-3 μM.

Example 11 Effect of JY0691, JY0692, JY0693, JY0694, JY0695 on Aortic Smooth Muscle Cell Proliferation Cycle

The changes in the cell cycle progression generated by treating the compounds of the present invention JY0691, JY0692, JY0693, JY0694, JY0695 to aortic smooth muscle cells induced by platelet-derived growth factor were examined. 4.0 x 10 ⁴ cells / ml of aortic smooth muscle cells were transferred to 6 well plates and incubated for 24 hours in DMEM medium containing 0.5% (V / V) fetal calf serum. Subsequently, the medium was treated with compounds JY0691, JY0692, JY0693, JY0694, JY0695 at concentrations of 0.5, 1, and 3 μM, and then further incubated for 24 hours, followed by platelet-dereived growth factor (PDGF). -BB) and cultured cells were stained using PI staining. Since the amount of the nucleus was confirmed by FACS (fluorescence activated cell sorter) method.

3A-3E show the effects of the compounds JY0691, JY0692, JY0693, JY0694, JY0695 on aortic smooth muscle cells of white rats at concentrations of 0.5 μM, 1 μM, and 3 μM, and then on the cell proliferation cycle of these cells. It is a graph. 3A to 3E show that the compounds JY0691, JY0692, JY0693, JY0694, and JY0695 of the present invention delay the inter-phase of G0 / G1 and S phase in the vascular smooth muscle cell cycle progression induced by platelet-derived growth factors. It was found that the cell proliferation was suppressed.

     Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (6)

The compound represented by following formula (I).       [Formula I] With the proviso that, in Formula I, R ₁ is C ₁ -C ₄ straight or branched alkyl. delete delete delete delete (a) a pharmaceutically effective amount of any one of the compounds represented by the following formulas (I), (II) and (III); And (b) vascular restenosis, vascular stenosis, arteriosclerosis, atherosclerosis, heart failure, myocardial infarction, angina pectoris, arrhythmia, hypertensive heart disease, congenital heart disease, stroke, including a pharmaceutically acceptable carrier, Or a pharmaceutical composition for preventing or treating peripheral vascular stenosis.       [Formula I] [Formula II]       [Formula III] Provided that in Formula (I), R ₁ is C ₁ -C ₄ straight or branched alkyl.