KR101210553B1 - Novel Glycolipid Derivative Compound and Composition For Treatment of Vascular Smooth Muscle Cell Hyper-Proliferation Comprising the Same - Google Patents

Abstract

The present invention provides a novel glycolipid derivative 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 abnormality proliferative disease comprising the compound as an active ingredient. To provide. Since the novel glycolipid derivative compounds of the present invention have excellent vascular smooth muscle cell proliferation inhibitory activity, vascular smooth muscle cell aberrant proliferative diseases such as atherosclerosis, atherosclerosis, vascular restenosis, and vascular stenosis as well as secondary to this It can be usefully used for the prevention or treatment of various cardiovascular diseases.

Description

Novel Glycolipid Derivative Compound and Composition For Treatment of Vascular Smooth Muscle Cell Hyper-Proliferation Comprising the Same

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

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 to promote the production and secretion of extracellular matrix (ECM), leading to thickening of the vascular endothelium 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 proliferative factor and activation of smooth muscle muscle factor, transmission of proliferation-induced signals through receptors on the surface of proliferative factor, and transfer into the nucleus of vascular smooth muscle cell. And cell cycle changes due to proliferation-induced signals.

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 proliferation of vascular smooth muscle cells is caused by various cytokines.

To date, antiplatelet agents, anticoagulants, cholesterol biosynthesis inhibitors and antiallergic agents have been used to prevent restenosis (Lefkovits et al., Progr. Cardiovas. Dis., 40, 141-158, 1997). Research has been conducted on the prevention of restenosis of natural oil-derived components such as fish oil, vitamin C and vitamin E. However, these drugs have not yet achieved 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. Korean Patent Registration No. 10-478671 discloses a pharmaceutical composition for preventing and treating blood restenosis comprising clotrimazole as an active ingredient, and Korean Patent Registration No. 10-516026 discloses 3'-deoxyadenosine (3 ' -Deoxyadenosine) is disclosed a composition for preventing and treating vascular restenosis as an active ingredient, Korean Patent Laid-Open No. 2001-110793 discloses an agent for preventing and treating vascular stenosis as an active ingredient a compound having Rho kinase inhibitory activity It is. In addition, Korean Patent Publication No. 2003-46314 discloses the prevention of vascular cell abnormality proliferative diseases such as transplanted vascular disease, restenosis, in-stent restenosis and pulmonary hypertension using antithrombin as an active ingredient and A therapeutic composition is disclosed, and Korean Patent Laid-Open Publication No. 2005-23249 discloses vascular diseases such as vascular restenosis and / or re-occluding after percutaneous coronary angioplasty using an intravascular stent containing retinoid and a retinoid action modifier. A pharmaceutical composition for prophylaxis and / or treatment is disclosed, and Korean Patent Laid-Open Publication No. 2005-43183 discloses a pharmaceutical composition for preventing and treating vascular restenosis including 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 suppressing vascular restenosis derived from the various proven natural products, no research results have been reported. Therefore, there is a continuing need to develop a substance that can safely and effectively inhibit vascular restenosis.

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 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.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have made diligent efforts to develop a substance that can effectively treat and prevent diseases caused by excessive proliferation of vascular smooth muscle cells. As a result, the glycolipid derivative compound synthesized by the present inventors is used for vascular smooth muscle cell (VSMC). The present invention was completed by experimentally confirming that there is a proliferation inhibitory activity.

Accordingly, an object of the present invention is to provide a novel glycolipid derivative compound.

Another object of the present invention to provide a composition for inhibiting vascular smooth muscle cell proliferation comprising a glycolipid derivative compound as an active ingredient.

Still another object of the present invention is to provide a pharmaceutical composition for preventing or treating vascular smooth muscle cell abnormal proliferative disease containing a glycolipid derivative compound as an active ingredient.

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

According to one aspect of the present invention, the present invention provides a novel glycolipid derivative compound represented by the following formula (1).

However, in the formula 1 R is E - cinnamoyl (E -cinnamoyl), C ₁ -C a C ₁ -C ₄ straight chain alkyl (alkyl) substituted by a straight-chain alkane ₁₀ Al (alkanal) or phenyl-triazol jolil (phenyl triazolyl) to be.

According to a preferred embodiment, the R in formula (I) is E - cinnamoyl _{(E -cinnamoyl), C 1 -C} 7 straight-chain alkane al (alkanal) or phenyl-triazol jolil a C ₁ substituted with a (phenyl triazolyl) -C ₃ straight alkyl.

According to a more preferred embodiment of the present invention, the compound of the present invention is a compound represented by the following formula (2), (3), (4) or (5).

The glycolipid derivative compound of Chemical Formula 1 is a novel compound synthesized by the inventors for the first time.

The compound of formula 2 (compound 2 ; 2a, 2b, 2c ), which is a specific compound of formula 1 of the present invention , may be synthesized by the following scheme 1.

[Reaction Scheme 1]

In Scheme 1, the reactant 'RCOH' is alkyl or aryl aldehyde, pyrrolidine is a base catalyst, and the solvent is anhydrous methylene chloride (M.C.).

Compounds used as starting materials in the above Reaction Scheme 1 1 (1a, 1b, 1c ), (2 R) -2- (2,4-Di- O -benzyl 3,6-dideoxy-α-L-rhamnopyranosyloxy) -octanone Silver can be easily prepared by modifying the terminal olefin group using oxygen and PdCl ₂ / CuCl in an olefinic glycolipid substituted with C ₁ -C ₄ linear alkyl. This reaction can be performed for 18 hours at room temperature from 0 ℃, using a reactor to suppress the inflow of air. Reactant aldehyde can be used in amounts of 1.0 equivalents relative to starting compound 1.

Compounds of Formulas 3 and 4 (Compounds 4 : 4a, 4b, 4c ), which are specific compounds of Formula 1 of the present invention, may be synthesized by the following Scheme 2.

Scheme 2

In the reaction scheme 2, the reactant 'R-≡' is ethynyl benzene, copper sulfate hydrate (CuSO ₄ ㆍ 5H ₂ O) and sodium ascorbate are catalysts, and the solvent is THF / H ₂ O (1: 1). Azide compounds used as starting materials in Scheme 2 (Compound 3 ; 3a, 3b, 3c ) [(2 R, 5 R) -2- (R) -6- azidohexan-2-yloxy) -6-methyl-tetrahydro-2 H -pyran-3,5-diol] is C ₁ -C ₄ straight chain alkyl ( It can be easily prepared by modifying the terminal olefin group from olefinic glycolipid substituted with alkyl). The reaction can be carried out at room temperature for 10 minutes to 4 hours, using a reactor to suppress the inflow of air. The reactant ethynylbenzene can be used in an amount of 1.0 equivalents to the starting compound 3 .

Specific Compound of Chemical Formula 1 Compound 6 of Chemical Formula 5 may be synthesized by the following Scheme 3.

Scheme 3

In order to obtain compound 6 in Scheme 3, starting compound 5 is a glycolipid aldehyde protected by two hydroxyl groups, and NaOMe is a base for removing benzoyl protecting group. Amberite IR-120 is an acid catalyst for neutralizing sodium salts, where the solvent is CH ₃ OH.

The aldehyde compound (Compound 5 ) [(2R) -2- (2,4-Di- O- benzyl-3,6-dideoxy-α-L-mannopyranosyloxy) tridecanal] used as a starting material in Scheme 3 is C ₁ It can be easily prepared by modifying the terminal olefin group from the olefinic glycolipid substituted with -C ₄ linear alkyl. The reaction can be carried out for 8 hours at room temperature, using a reactor to suppress the inflow of air. Protective group remover NaOMe can be used in an amount of 3.0 equivalents relative to starting compound 5 .

According to another aspect of the invention, the present invention provides a composition for inhibiting vascular smooth muscle cell proliferation comprising the glycolipid derivative compound represented by the formula (1).

According to a preferred embodiment, the active ingredient compound of the present invention composition is that the R in the general formula E 1 - cinnamoyl _{(E -cinnamoyl), C 1 -C} 7 straight-chain alkane al (alkanal) or phenyl-triazol jolil ( phenyltriazolyl) substituted with C ₁ -C ₃ straight alkyl.

According to a more preferred embodiment of the present invention, the active ingredient compound of the present invention is a compound represented by the formula (2), (3), (4) or (5).

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

According to another aspect of the present invention, the present invention provides a vascular smooth muscle cell abnormal proliferative disease comprising (a) a therapeutically effective amount of a glycolipid derivative compound represented by the formula (1) and (b) a pharmaceutically acceptable carrier ( Provided is a pharmaceutical composition for preventing or treating vascular smooth muscle cell hyper-proliferative disorder.

Glycolipid derivative compounds of the present invention have 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 synthesis of DNA at the time of PDGF-BB-induced aortic smooth muscle cell proliferation of white rats, thereby inhibiting the proliferation of this cell.

As used herein, the term "smooth muscle" refers to the walls of organs such as tubular organs, bladder, abdominal cavity, uterus, reproductive tract, gastrointestinal tract, respiratory tract, vasculature, skin and ciliary muscle, eye iris, etc. It is a non-striated muscle found in. 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).

As used herein, the term “vascular smooth muscle cell (VSMC)” refers to the cells that make up the smooth muscle of the vessel wall.

As used herein, the term "VSMC hyper-proliferative disorder" refers to a disease or condition resulting from excessive proliferation of vascular smooth muscle cells.

In the present invention, "vascular smooth muscle cell aberrant proliferative disease" refers to vascular stenosis, vascular stenosis, arteriosclerosis, atherosclerosis directly caused by abnormal proliferation of vascular smooth muscle cells, as well as vascular stenosis, vascular stenosis or artery Cardiovascular diseases secondary to sclerosis, including heart failure, myocardial infarction, angina pectoris, arrhythmia, hypertensive heart disease, congenital heart disease, stroke, and peripheral vascular stenosis. More 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 the arteries. On the other hand, restenosis of the blood vessel is a disease in which the vascular passage is narrowed after the vessel wall is traumatized. 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 prevent vascular restenosis, studies have been conducted on drugs that inhibit vascular smooth muscle cell proliferation, and several drugs are now used in the treatment of patients (J. Am. Coll. Cardiol., 2002, 39, 183-193). Therefore, the glycolipid derivative compounds of the present invention which effectively inhibit the proliferation of vascular smooth muscle cells can be effectively used for the treatment of vascular smooth muscle cell abnormal proliferative diseases.

The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient.

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).

The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, .

The pharmaceutical composition of the present invention may be administered orally or parenterally, and when administered parenterally, may be administered by intravenous infusion, subcutaneous injection, 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 glycolipid derivative compound which is an active ingredient included in the composition of the present invention may be determined in consideration of the purpose of treatment, the condition of the patient, the period of time required, and the like, and is not limited to a specific range of concentration. Preferably, the glycolipid derivative compound of the present invention may be added in the composition at a concentration of 5-50 μM, more preferably at a concentration of 10-50 μM, even more preferably at 20-50 μM. If it is contained in a concentration of less than 5μM it is difficult to expect the desired effect, if it is contained in a concentration of more than 50μM it is difficult to expect the effect of increasing in proportion to the increase in the concentration, the manufacturing cost of the drug prepared using the compound of the present invention This has an increasing disadvantage.

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. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The present invention provides a novel glycolipid derivative 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 abnormality proliferative disease comprising the compound as an active ingredient. To provide. Since the novel glycolipid derivative compounds of the present invention have excellent vascular smooth muscle cell proliferation inhibitory activity, vascular smooth muscle cell aberrant proliferative diseases such as atherosclerosis, atherosclerosis, vascular restenosis, and vascular stenosis as well as secondary to this It can be usefully used for the prevention or treatment of various cardiovascular diseases.

Figures 1a to 1c shows the results of measuring the inhibition of proliferation of aortic vascular smooth muscle cells by the present invention glycolipid derivatives of various concentrations.
Figure 1a is treated with the platelet-derived growth factor (PDGF-BB) after the treatment of the aortic vascular smooth muscle cells of white rats with the glycolipide derivative compound DD189 of the present invention represented by the formula (2) at a concentration of 10, 20, 40 μM Induced to show the result of measuring the extent to which the proliferation of this cell is inhibited.
Figure 1b is treated with a platelet-derived growth factor (PDGF-BB) after treating the glycolipid derivative compound DD204 of the present invention represented by the formula (3) to aortic vascular smooth muscle cells of white rats at a concentration of 10, 20, 50 μM Induced to show the result of measuring the extent of inhibition of proliferation of this cell.
Figure 1c is treated with a platelet-derived growth factor (PDGF-BB) after treating the glycolipide derivative compound DD223 of the present invention represented by the formula (4) to aortic vascular smooth muscle cells of white rats at a concentration of 5, 20, 50 μM Induced to show the result of measuring the extent to which the proliferation of this cell is inhibited.
2a to 2c shows the results of measuring the DNA synthesis inhibitory effect of aortic vascular smooth muscle cells by the present invention glycolipid derivatives of various concentrations.
Figure 2a is treated with white glycolipide derivative compound DD189 of the present invention represented by the formula (2) at a concentration of 10, 20, 40μM to white rat aortic vascular smooth muscle cells, and then platelet-derived growth factor (PDGF-BB) Induced and measured the extent to which the DNA synthesis of these cells is inhibited.
Figure 2b is treated with white glycolipid derivative compound DD204 of the present invention represented by the formula (3) at a concentration of 10, 20, 50μM to white rat aortic vascular smooth muscle cells, and then platelet-derived growth factor (PDGF-BB) Induced and measured the extent to which the DNA synthesis of these cells is inhibited.
Figure 2c is treated with a glycolipide derivative compound DD223 of the present invention represented by the formula (4) in aortic vascular smooth muscle cells of white rats at concentrations of 5, 20, 50μM, and then proliferated by platelet-derived growth factor (PDGF-BB) Induced and measured the extent to which the DNA synthesis of these cells is inhibited.
Figure 3a to 3c is a glycolipid derivative of the present invention at various concentrations, the inter-phase of the G0 / G1 and S phase of the cell growth cycle of aortic vascular smooth muscle cells is delayed, showing the effect of inhibiting cell proliferation Shows that
Figure 3a is treated with white glycolipide derivative compound DD189 of the present invention represented by the formula (2) at a concentration of 10, 20, 40μM to aortic vascular smooth muscle cells of the rat, platelet-derived growth factor (PDGF-BB) Induction and measurement of the cell proliferation cycle of these cells are shown.
FIG. 3b shows that the glycolipide derivative compound DD204 of the present invention represented by Chemical Formula 3 is treated with aortic vascular smooth muscle cells of white rats at concentrations of 10, 20, and 50 μM, and then proliferated by platelet-derived growth factor (PDGF-BB). Induction and measurement of the cell proliferation cycle of these cells are shown.
FIG. 3C shows that the glycolipide derivative compound DD223 of the present invention represented by Chemical Formula 4 is treated with aortic vascular smooth muscle cells of white rats at concentrations of 5, 20, and 50 μM, followed by proliferation by platelet-derived growth factor (PDGF-BB). Induction and measurement of the cell proliferation cycle of these cells are shown.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Example  One : ( E ) -1- (3,6- dideoxy -α-L- rhamnosyloxy ) -9- phenylnon -8- en -7- one  [ chemistry Synthesis of Compound of Formula 2]

[Formula 2]

Round bottom flask (round flask) Compound 1, (2 R) -2- ( 2,4-Di- O -benzyl 3,6-dideoxy-α-L-rhamnopyranosyl -oxy) -octanone (47mg, 0.08mmol) in the It was added and dissolved in 7 mL of MeOH. The temperature was lowered and NaOCH ₃ (6 mg, 0.25 mmol) was slowly added at 0 ° C. As the benzoyl protecting group was removed, the solution became more yellowish. After stirring for 12 hours, the reaction was terminated and added Amberlite-IR120 (H ⁺ ) little by little directly in MeOH solvent to check the neutrality of the pH paper. When neutral, the resin was filtered to remove the resin, the solvent was removed by distillation under reduced pressure, and then extracted with distilled water and EtOAc (5 mL X 3). MgSO ₄ , dried (filtered), dried (filtered) and concentrated under reduced pressure, using flash column chromatography (CH ₂ Cl ₂ / MeOH, 15: 1, v / v) Compound 2 (Formula 2 ) 2 compound) (27 mg, yield 88%) was isolated. Physical and chemical properties of the isolated compound were as follows.

C ₂₁ H ₃₀ O _{5 ,} colorless syrup, Rf = 0.56 (CH ₂ Cl ₂ / MeOH, 7: 1, v / v); [?] _D ²⁰ = -59.17 ° (c = 0.06, CHCl ₃ ); ¹ H-NMR (250 MHz, MeOD) δ 7.59-7.53 (m, 3H), 7.41-7.38 (m, 2H), 7.33-7.32 (m, 1H), 6.77-6.71 (d, J = 15 Hz, 1H) , 4.72 (s, 1H) 3.81 (m, 2H), 3.75-3.64 (m, 1H), 3.62-3.48 (m, 1H), 2.70 (t, 2H), 2.13-2.05 (m, 1H), 1.92- 1.77 (m, 1H), 1.74-1.66 (m, 2H), 1.64-1.43 (m, 4H), 1.30-1.27 (d, J = 7.5 Hz, 3H), 1.14-1.12 (d, J = 5 Hz, 3H); ¹³ C-NMR (62.9 MHz, MeOD); δ 200.88, 142.85, 134.59, 130.62, 129.07, 128.70, 128.42, 126.65, 126.32, 95.90, 71.02, 69.98, 69.42, 68.19, 40.82, 37.11, 35.57, 25.49, 24.44, 19.05, 17.87; IR (film) v _max 3417, 2963, 2929, 2872, 1656, 1609, 1450, 1408, 1384, 1242, 1124, 1098, 1028, 984 cm ⁻¹ ; LC Mass ESI probe 385 Da / e, Scan ES + 5.61e

Example  2: (3 R , 5 R , 6 R )-2- methyl -6-(( R ) -7- (4- phenyl -One H -1,2,3- triazole -One- yl ) heptan-2-yloxy) -tetrahydro-2 H Synthesis of -pyran-3,5-diol [Compound of Formula 3].

(3)

Azide compound 3 (n = 3), ( 2 R, 5 R) -2- (R) -6-azidohexan-2-yloxy) -6-methyl- tetrahydro -2 H -pyran-3,5-diol ( CuSO ₄ ㆍ 5H ₂ O (1.5 mg, 0.006 mmol) in THF / H ₂ O (1: 1) (5 mL) solution with 16 mg, 0.06 mmol) and ethynylbenzene (6 mg, 0.06 mmol, 1eq) And sodium ascorbate (2.4 mg, 0.012 mmol) were added. The mixture was stirred at room temperature for 10 minutes to confirm the reaction by TLC. When the reaction was completed, the mixture was diluted with ethyl acetate (20 ml), washed with brine, dried over MgSO ₄ , filtered, and concentrated under reduced pressure. Pure compound 4 (compound of formula 3) (19 mg, 90% yield) was isolated using flash column chromatography (CH ₂ Cl ₂ / MeOH, 15: 1, v / v). Physical and chemical properties of the isolated compound were as follows.

C ₂₁ H ₃₁ N ₃ O ₄ , white solid, R _f = 0.3 (MC / MeOH, 9: 1, v / v); [a] _D ² ° = -88.00 ° (c = 0.03, MeOH); ¹ H-NMR (250 MHz, CDCl3) δ 7.83-7.80 (d, J = 7.5 Hz, 2H), 7.76 (s, 1H), 7.44-7.40 (t, 2H), 7.34-7.31 (t, 1H), 4.69 (s, 1H), 4.44-4.37 (t, 2H), 3.79-3.75 (m, 2H), 2.17 (s, 2H), 2.1-2.04 (m, 1H), 2.00-1.93 (m, 2H), 1.86 -1.83 (m, 1H), 1.56-1.35 (m, 6H), 1.26-1.24 (d, J = 5 Hz, 3H), 1.11-1.09 (d, J = 5 Hz, 3H) ¹³ C-NMR (62.9 MHz, CDCl3) δ 147.85, 130.56, 128.97, 128.30, 125.81, 119.63, 96.17, 71.21, 70.08, 69.41, 68.09, 50.35, 37.05, 35.37, 31.03, 30.20, 26.60, 24.84, 19.04, 17.86. IR (film) vmax 3392, 2929, 1739, 1650, 1450, 1383, 1228, 1125, 1099, 1043, 1028, 982, 833, 766, 695 cm ^-1 LC Mass ESI probe 412 Da / e, Scan ES + 5.7e

Example 3: (3 R , 5 R , 6 R ) -2-methyl-6-(( R ) -8- (4-phenyl-1 H -1,2,3-triazol-1-yl) octan-2-yloxy) -tetrahydro-2 H Synthesis of -pyran-3,5-diol [Compound of Formula 4].

[Formula 4]

Azide Compound 3 (n = 4), (2 R , 5 R ) -2-(( R ) -8-azidooctan-2-yloxy) -6-methyl-tetrahydro-2 H -pyran-3,5-diol (15 mg, 0.04 mmol) and THF / H ₂ O (1: 1) (5 mL) solution with ethynylbenzene (6 mg, 0.06 mmol, 1 eq) dissolved in CuSO ₄ .5H ₂ O (1.5 mg, 0.006 mmol) and sodium ascorbate (2.4 mg, 0.012 mmol) were added. The mixture was stirred at room temperature for 10 minutes to confirm the reaction by TLC. When the reaction was completed, the mixture was diluted with ethyl acetate (20 mL), washed with brine, dried over MgSO ₄ , filtered, and concentrated under reduced pressure. Pure compound 4 (compound of formula 4) (11 mg, yield 92%) was isolated using flash column chromatography (CH ₂ Cl ₂ / MeOH, 15: 1, v / v). The physical and chemical properties of the isolated compound were as follows.

C ₂₂ H ₃₃ N ₃ O ₄ , white solid, R _f = 0.3 (MC / MeOH, 9: 1, v / v); [a] _D ² ° = -86.00 ° (c = 0.05, MeOH); ¹ H-NMR (250MHz, MeOD) δ 8.32 (s, 1H), 7.83-7.80 (d, J = 7.5Hz, 2H), 7.45-7.30 (m, 3H), 4.63 (s, 1H), 4.47-4.41 (t, 2H), 3.77-3.68 (m, 2H), 3.65-3.46 (m, 2H), 1.98-1.91 (m, 3H), 1.81-1.70 (m, 1H), 1.6-1.1.32 (m, 8H), 1.21-1.18 (d, J = 7.5 Hz, 3H), 1.11-1.09 (d, J = 5 Hz, 3H); ¹³ C-NMR (62.9 MHz, MeOD) δ 148.83, 131.75, 129.98, 129.32, 126.66, 122.177, 97.55, 72.49, 71.17, 69.94, 68.30, 51.48, 38.23, 35.95, 31.24, 30.03, 27.44, 26.66, 19.36, 18.12. IR (film) vmax 3395, 2939, 1734, 1645, 1634, 1450, 1383, 1228, 1125, 1099, 1043, 1028, 982, 833, 726, 678 cm ^-1 LC Mass ESI probe 426 Da / e, Scan ES + 4.7e

Example  4: (2 R ) -2- (3,6- dideoxy -α-L- mannopyranosyloxy ) - tridecanal  Synthesis of [Compound 5].

[Chemical Formula 5]

Compound To a round flask (2 R) -2- (2,4- Di- O -benzyl-3,6-dideoxy-α-L-mannopyranosyoxy) into the -tridecanal (40mg, 0.07mmol) was dissolved in MeOH 5㎖. Lower the temperature to NaOCH ₃ at 0 ° C. (14 mg, 0.21 mmol) was added and the temperature was raised to room temperature. As the benzoyl protecting group was removed, the solution became more yellowish. After stirring for 8 hours, the reaction was terminated, and then added Amberlite-IR120 (H ⁺ ) little by little in the MeOH solvent to check the neutrality of the pH paper (paper). When neutral, the resin is filtered to remove the resin, the solvent is removed by distillation under reduced pressure, and then extracted with distilled water and EtOAc (20 mL X 3). The organic layer was washed with brine, dried over MgSO ₄ , filtered, concentrated under reduced pressure, and subjected to flash column chromatography to compound 6 (compound 5) (19.5). mg, yield 86%) was obtained. The physical and chemical properties of the isolated compound were as follows.

C ₁₉ H ₃₆ O ₅ , a colorless syrup, Rf = 0.22 (n-Hexane / EtOAc / MeOH, 5: 2: 1, v / v / v); [a] _D ²³ = -11.0 (c = 0.05, MeOH); ¹ H NMR (250 MHz, CDCl ₃ ) δ 9.68 (s, 1H), 4.61 (s, 1H). 3.73-3.67 (m, 2H), 3.64-3.47 (m, 2H), 2.63 (bs, 2H), 2.38-2.32 (m, 2H), 2.00-1.95 (m, 1H), 1.80-1.71 (m, 1H ), 1.58-1.43 (m, 4H), 1.42-1.18 (m, 17H), 1.05 (d, 3H, J = 5.9 Hz), 1.14 (d, 3H, J = 6.1 Hz); ¹³ C NMR (62.9 MHz, CDCl ₃ ) δ 203.1, 96.1, 71.6, 69.8, 69.3, 67.9, 43.9, 37.2, 35.1, 29.6, 29.4, 29.3, 29.1, 25.8, 22.1, 19.0, 17.7; IR (film) v _max 3430, 2927, 2854, 1636, 1558, 1457, 1377, 1253, 1123, 1028 cm ^-1 .

Example  5: aorta Vascular smooth muscle cell  Effect on growth inhibition DD189 , DD204 , DD223 Effect

Whether the glycolipid derivative compounds of the present invention (hereinafter, referred to as DD189, DD204, and DD223, respectively) represented by Formulas 2, 3, and 4 synthesized in the above examples inhibit the growth of aortic vascular smooth muscle cells in white rats. Confirmed. First, 4.0 × 10 ⁴ cells / ml of aortic vascular smooth muscle cells were transferred to 12-well plates and incubated for 24 hours in DMEM medium containing 0.5% (V / V) fetal bovine serum. Subsequently, the medium was treated with compound DD189 at a concentration of 10 μM, 20 μM, 40 μM, compound DD204 at a concentration of 10 μM, 20 μM, 50 μM, and compound DD223 at a concentration of 5 μM, 20 μM, 50 μM, respectively, and then again for 24 hours. After further incubation, the cells were treated with platelet-dereived growth factor (PDGF-BB), and after 24 hours with trypsin to separate the cells from the bottom and then separated using a hemocytometer. The number of cells was counted.

1A-1C, after treatment with aortic smooth muscle cells of white rats with compounds DD189 (concentrations 10, 20, 40 μM), DD204 (concentrations 10, 20, 50 μM), and DD223 (concentrations 5, 20, 50 μM), The graph shows the result of measuring the extent to which the proliferation of these cells is inhibited. As shown in the results of FIGS. 1A to 1C, it was confirmed that the glycolipid derivative compounds of the present invention, DD189, DD204, and DD223 inhibit the proliferation of aortic vascular smooth muscle cells in a concentration-dependent manner.

Example  6: compound DD189 , DD204 , DD223 Aortic Smooth Muscle Cells DNA  Synthesis impact

In Example 5, the compounds of the present invention, DD189, DD204 and DD223 was confirmed to inhibit the proliferation of aortic vascular smooth muscle cells in white rats. In order to determine the mechanism by which DD189, DD204, and DD223 compounds inhibit the proliferation of aortic vascular smooth muscle cells, the effects of the compounds DD189, DD204, and DD223 on cell proliferation were examined. First, 2.0 × 10 ⁴ cells / ml of aortic vascular smooth muscle cells were transferred to a 24-well plate and incubated for 24 hours in DMEM medium containing 0.5% (V / V) fetal bovine serum. Subsequently, the medium was treated with compound DD189 at a concentration of 10, 20, 40 μM, compound DD204 at a concentration of 10, 20, 50 μM, and compound DD223 at a concentration of 5, 20, 50 μM, respectively, followed by another 24 hours. After incubation, it was treated with platelet-dereived growth factor (PDGF-BB). After 20 hours, 1 μL of [ ³ H] -thymidine was added. After 4 hours of incubation, the cells were 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 with a liquid scintillation counter.

2A to 2C show that the rat aortic vascular smooth muscle cells were treated with the compounds DD189 (10, 20, 40 μM), DD204 (10, 20, 50 μM) and DD223 (5, 20, 50 μM). The graph shows the results of measuring the degree of inhibition of DNA synthesis of cells. From the results shown in FIGS. 2A, 2B and 2C, the compounds DD189 (10, 20, 40 μM), DD204 (10, 20, 50 μM), and DD223 (5, 20, 50 μM) of the present invention were concentration dependent. It was found that the biosynthesis of DNA of aortic vascular smooth muscle cells was effectively inhibited.

Example 7 Effect of Compounds DD189, DD204, and DD223 on Aortic Vascular Smooth Muscle Cell Proliferation Cycle

The change in cell cycle progression generated by treatment of the compounds DD189, DD204, and DD223 of the present invention on aortic vascular smooth muscle cells inducing proliferation with platelet-derived growth factors was investigated. First, 4.0 × 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 Compound DD189 at a concentration of 10, 20, 40 μM, Compound DD204 at a concentration of 10, 20, 50 μM, and Compound DD223 at a concentration of 5, 20, 50 μM, followed by another 24 hours. The cells were further cultured, then treated with platelet-dereived growth factor (PDGF-BB), and further cultured cells were stained using PI staining. Since the amount of the nucleus was confirmed by FACS (fluorescence activated cell sorter) method.

3A, 3B, and 3C show the rat aorta by treatment with Compound DD189 (10, 20, 40 μM), Compound DD204 (10, 20, 50 μM), and Compound DD223 (5, 20, 50 μM). The results of measuring the effect on the cell proliferation cycle of vascular smooth muscle cells are shown graphically. From the results shown in FIGS. 3A, 3B, and 3C, compounds DD189, DD204, and DD223 are inter-faces of G0 / G1 and S phases in the cycle progression of aortic vascular smooth muscle cells induced by platelet-derived growth factors. retardation of cell proliferation.

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. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (6)

Glycolipid derivative compound represented by the following formula (1).
[Formula 1]

However, in the above formula 1, R is E - a cinnamoyl (E -cinnamoyl) or phenyl-triazol jolil a C ₁ -C ₄ straight chain alkyl (alkyl) substituted with (phenyltriazolyl).
A composition for inhibiting vascular smooth muscle cell proliferation, including a glycolipid derivative compound represented by Formula 1 below.
[Formula 1]

However, in the above formula 1, R is E - cinnamoyl _{(E -cinnamoyl), C 1 -C} 10 straight chain alkane al (alkanal) or phenyl-triazol jolil (phenyl triazolyl) a C ₁ -C ₄ straight chain alkyl (substituted by alkyl )to be.
(a) a therapeutically effective amount of a glycolipid derivative compound represented by Formula 1 below; And (b) a pharmaceutical composition for preventing or treating vascular smooth muscle cell aberrant proliferative disease comprising a pharmaceutically acceptable carrier.
[Formula 1]

However, in the above formula 1, R is E - cinnamoyl (E -cinnamoyl), C ₁ -C a C ₁ -C ₄ straight chain alkyl (alkyl) substituted by a straight-chain alkane ₁₀ Al (alkanal) or phenyl-triazol jolil (phenyltriazolyl) to be.
4. The composition of claim 3, wherein the glycolipid derivative compound of Formula 1 is a compound of Formula 2, Formula 3, Formula 4 or Formula 5.
(2)

(3)

[Chemical Formula 4]

[Chemical Formula 5]

The method of claim 3, wherein the vascular smooth muscle cell aberrant proliferative disease includes vascular restenosis, vascular stenosis, arteriosclerosis, atherosclerosis, heart failure, myocardial infarction, angina pectoris, arrhythmia, hypertensive heart disease, congenital heart disease, Stroke, or peripheral vascular stenosis.
The composition of claim 3, wherein the compound of Formula 1 is included at a concentration of 5-50 μM relative to the composition.

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