KR20160083806A - Composition for treating and preventing of cholesterol related to disease - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of cholesterol-related diseases, and more particularly to a composition for preventing and treating cholesterol-related diseases, which contains a compound inhibiting the binding of PCSK9 and LDLR as an active ingredient, Or the LDLR expression level is not directly controlled, thereby lowering the side effects. It is anticipated that the combination of PCSK9 and LDLR will effectively inhibit plasma total cholesterol levels, while at the same time increasing the therapeutic efficacy of statin drugs in combination with statins.

Description

[0001] The present invention relates to a composition for preventing and treating cholesterol-related diseases,

The present invention relates to a composition for preventing and treating cholesterol-related diseases.

Cholesterol is a steroid compound that is widely used as a cellular component of higher animals and is one of the essential nutrients in our body. However, recent westernized dietary habits and changes in living conditions have raised the level of cholesterol in blood abnormally, resulting in an increase in the incidence of cholesterol-related diseases such as dyslipidemia, heart disease, diabetes and cardiovascular disease, and development of drugs to lower blood cholesterol . Currently, there are two ways to reduce blood cholesterol levels. The first is to directly reduce the amount of cholesterol produced in the liver and the second is to increase the amount of low density lipoprotein receptor (LDLR), which causes blood cholesterol to enter the cells, Cholesterol is metabolized and excreted in the liver, which ultimately reduces blood cholesterol levels.

The most commonly used drug that inhibits the production of cholesterol in the liver is the statin-based drug. It is known that statins inhibit the activity of HMG-CoA reductase involved in the synthesis of cholesterol, thereby decreasing the cholesterol concentration in the blood and secondarily enhancing the expression of the low density lipoprotein receptor in the cell membrane have. However, these statins are known to show side effects such as muscle pain due to muscle toxicity, diabetes mellitus due to increased blood sugar, increase in neurological abnormalities such as confusion and confusion, and the cholesterol concentration is not controlled even with statin administration As the number of patients is increasing, it is necessary to develop drugs that can lower new blood cholesterol levels.

On the other hand, the pro-protein converting enzyme subtilisin / kexin type 9 (PCSK9) protein binds to the EGF-A domain of the low density lipoprotein receptor present in the plasma membrane of the liver, It has been shown that the receptor is degraded by lysosomes and results in a quantitative reduction of low density lipoprotein receptors and enhances the blood cholesterol concentration. There have been active attempts to use PCSK9 inhibitors to lower blood cholesterol levels (Trends in Biochemical Sciences (2007) 32: 71-77).

In addition, statin-based drugs are known to increase the expression of PCSK9, and the reason why the LDL-cholesterol-lowering effect of statins does not increase in proportion to the administration dose is that PCSK9 . This suggests that the combination of PCSK9 inhibitor and statin drugs may increase the therapeutic efficacy. Therefore, the development of PCSK9 inhibitors is becoming increasingly popular. However, there is a continuing controversy over the development of other side effects of drugs that directly inhibit the expression of PCSK9 in the most recently developed liver.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made in order to solve the above problems of the prior art, and it is an object of the present invention to provide a method for screening for a compound that inhibits the binding of PCSK9 and LDLR, It is another object of the present invention to provide a composition for preventing and treating diseases.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions, and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and techniques of manufacture are not described in any detail, in order not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "embodiment" means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Accordingly, the appearances of the phrase " in one embodiment "or" an embodiment "in various places throughout this specification are not necessarily indicative of the same embodiment of the present invention. In addition, a particular feature, form, composition, or characteristic may be combined in any suitable manner in one or more embodiments.

The present invention provides a composition for preventing and treating cholesterol-related diseases, which comprises a compound represented by the following formula (1) as an active ingredient.

The present invention also provides a composition for inhibiting PCSK9 and LDLR binding comprising a compound represented by the following formula (1) as an active ingredient.

The present invention also provides a composition for the prevention and treatment of cholesterol-related diseases, comprising a statin compound and a compound represented by the following formula (1) as an active ingredient.

[Chemical Formula 1]

R is _{N (CH 3) 2, N} (CH 2) 4 NH, CH 2 NH (CH 2) 3 OCH 3, CH 2 NH (CH 2) 2 N (CH 3) 2, NH (CH 2) 3 or O (CH ₂ ) ₂ NH (CH ₂ ) ₂ N (CH ₃ ) ₂ .

In one embodiment of the present invention, the compound represented by the formula (1) is preferably one or more compounds represented by the following formulas (2) to (7).

(2)

(3)

[Chemical Formula 4]

[Chemical Formula 5]

[Chemical Formula 6]

(7)

The present invention also provides a composition for preventing and treating cholesterol-related diseases, which comprises a compound represented by the following formula (8) as an active ingredient.

The present invention also provides PCSK9 and a composition for inhibiting LDLR binding comprising a compound represented by the following general formula (8) as an active ingredient.

The present invention also provides a composition for the prevention and treatment of cholesterol-related diseases, which comprises a statin compound and a compound represented by the following formula (8) as an active ingredient.

[Chemical Formula 8]

R ₁ is any one of H, Cl, or CH ₃ , R ₂ is H, OCH ₃ Or _{O (CH 2) 4 NH (} CH 2) 2 N (CH 2) 4 O either and, R ₃ is _{H, CH 2 N (CH 2} ) 4 CHCON (CH 2) 2 CH 3 C (CH) of _{5, CH 2 NHCH 2 C (} CH) 3 O , or _{CH 2 N (CH 2) 4} CHCON (CH 2) and any one of _4, R ₄ is _{CH 2 N (CH 2) 3} C 2 (CH) 2 C _{2 (OCH 3) 2, CH} 2 N (CH 2) 4 CHCONH (CH 2) 2 N (CH 2 CH 3) 2, or _{CH 2 NH (CH 2) 2} N (CH 2) is any one of ₄ O.

In one embodiment of the present invention, the compound represented by the formula (8) is preferably at least one of the compounds represented by the following formulas (9) to (16).

[Chemical Formula 9]

[Chemical formula 10]

(11)

[Chemical Formula 12]

[Chemical Formula 13]

[Chemical Formula 14]

[Chemical Formula 15]

[Chemical Formula 16]

The present invention also provides a composition for preventing and treating cholesterol-related diseases, which comprises a compound represented by the following general formula (17) as an active ingredient.

The present invention also provides a composition for inhibiting PCSK9 and LDLR binding comprising a compound represented by the following formula (17) as an active ingredient.

The present invention also provides a composition for the prevention and treatment of cholesterol-related diseases, comprising a statin compound and a compound represented by the following formula (17) as an active ingredient.

[Chemical Formula 17]

R ₁ is H or O (CH ₂ ) ₂ O (CH ₂ ) ₂ NH (CH ₂ ) ₂ N (CH ₃ ) ₂ , R ₂ is H and R ₃ is H, OCH ₂ CON ₂ ) ₄ CHN (CH ₂ ) ₄ NCH ₂ CH ₃ or O (CH ₂ ) ₃ NH (CH ₂ ) ₂ N (CH ₂ ) ₄ O and R ₄ is H.

In one embodiment of the present invention, the compound represented by the general formula (17) is preferably one or more of the compounds represented by the following general formulas (19) to (21).

[Chemical Formula 19]

[Chemical Formula 20]

[Chemical Formula 21]

The present invention also provides a composition for the prevention and treatment of cholesterol-related diseases comprising as an active ingredient any one or more of the compounds represented by the following formulas (18), (22), (23), (24), (25)

In addition, the present invention provides PCSK9 and a composition for inhibiting LDLR binding comprising at least one of the compounds represented by the following formulas (18), (22), (23), (24), (25)

The present invention also relates to a composition for prevention and treatment of cholesterol-related diseases comprising as an active ingredient any one or more of a statin compound and a compound represented by the following general formula (18), (22), (23), .

[Chemical Formula 18]

[Chemical Formula 22]

(23)

≪ EMI ID =

(25)

(26)

In another embodiment of the present invention, the cholesterol-related disorder is preferably a metabolic disease, a dyslipidemia disease, or the like. The metabolic diseases may preferably be obesity, diabetes, hypertension, cardiovascular disease and the like. The dyslipidemic disease is a disease in which total cholesterol, low density lipoprotein-cholesterol LDL-cholesterol, or triglycerides, preferably hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, and the like. However, the present invention is not limited to a disease that can be caused by an increase in blood cholesterol level (including plasma cholesterol) and / or an increase in LDL-cholesterol.

In another embodiment of the present invention, the statin compound is preferably selected from the group consisting of lovastatin, pravastatin, simvastatin, fluvastatin, atorvastatin, cerivastatin, rosuvastatin, and the like, but it is not limited to the HMG-CoA reductase inhibitor.

In the present invention, the composition may be in the form of capsules, tablets, granules, injections, ointments, powders or beverages, and the composition may be a human. The composition may be formulated in the form of oral preparations such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories and sterilized injection solutions according to a conventional method. The composition of the present invention may comprise a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be a binder, a lubricant, a disintegrant, an excipient, a solubilizing agent, a dispersing agent, a stabilizer, a suspending agent, a coloring matter, a perfume or the like in the case of oral administration. A solubilizing agent, an isotonic agent, a stabilizer and the like may be mixed and used. In the case of topical administration, a base, an excipient, a lubricant, a preservative and the like may be used. Formulations of the pharmaceutical compositions of the present invention may be prepared in a variety of ways by mixing with pharmaceutically acceptable carriers as described above. For example, oral administration may be in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc. In the case of injections, they may be formulated in unit dosage ampoules or in multiple dosage forms have. Other, solutions, suspensions, tablets, capsules, sustained-release preparations and the like.

Examples of suitable carriers, excipients and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltoditol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil. Further, it may further include a filler, an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, an antiseptic, and the like.

The route of administration of the composition according to the present invention may be, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, Or workplace. Oral or parenteral administration is preferred. The term "parenteral" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical compositions of the present invention may also be administered in the form of suppositories for rectal administration.

The compositions of the present invention may be administered in a variety of ways depending on various factors including the activity, age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and severity of the particular disease to be prevented or treated, And the dosage of the composition varies depending on the condition of the patient, the body weight, the degree of disease, the type of drug, the route of administration, and the period of time, but may be suitably selected by those skilled in the art and is preferably from 0.0001 to 50 mg / 50 mg / kg < / RTI > The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way. The pharmaceutical composition according to the present invention can be formulated into pills, dragees, capsules, solutions, gels, syrups, slurries, and suspensions.

The composition for the prevention and treatment of cholesterol-related diseases according to the present invention can not directly control the expression level of PCSK9 and / or LDLR in the liver, thereby lowering side effects thereof. It is anticipated that the combination of PCSK9 and LDLR will effectively inhibit plasma total cholesterol levels, while at the same time increasing the therapeutic efficacy of statin drugs in combination with statins.

1 is a graph showing the results of in vitro experiments according to an embodiment of the present invention.
2 is a graph showing the results of an in vivo experiment according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to 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: PCSK9 - LDLR  Screening of binding inhibitors

1.1. PCSK9 - LDLR  Selection of binding inhibition compounds

Compounds that are expected to inhibit the interaction of the pro-protein translocation enzyme subtilisin / kexin type 9, PCSK9 protein with low density lipoprotein receptor (LDLR) , The tertiary crystal structure of the PCSK9 protein bound to the EGF-A domain of LDLR at neutral pH was first determined using the Protein Data Bank. The site of the PCSK9 protein binding to LDLR was set to the region from the 367th amino acid to the 381st amino acid of the PCSK9 protein (Proceedings of the National Academy of Science (2008) 105: 1820-1825). Thereafter, the EGF-A domain was removed from the crystal structure, and the crystal structure of PCSK9 in which water molecules and hydrogen atoms were removed was regenerated again. The docking score of the 450,000 compounds in the ChemBridge Express collection and the crystal structure of the regenerated PCSK9 were measured using GOLD software (version 4.0.1) (Proteins (2003) 52 ): 609-623). In the measurement, up to 10 binding sites (docked poses) per compound were measured so as to confirm binding values with 200% searching efficiency. The top 100 compounds with the highest measured binding values were selected, and the selected compounds were purchased from ChemBridge.

1.2. PCSK9 - LDLR  Binding inhibition experiment

In order to confirm that 100 compounds selected in Example 1.1 inhibited PCSK9-LDLR binding, PCSK9-LDLR binding inhibition experiment was performed using CircuLex PCSK9-LDLR in vitro binding assay kit (MBL international). 10 ug of each compound and 100 ng of His-tagged PCSK9 were reacted for 1 hour at room temperature in a volume of 100 uL, transferred to an ELISA plate coated with an EGF-AB peptide of LDLR, and reacted for 1 hour. After the unbound His-Tagged PCSK9 was washed and removed, the antibody solution against the His-tag was reacted according to the manufacturer's method, and the binding rate of the EGF-AB peptide in the His-tag and the ELISA plate was confirmed by adding a color- Relative inhibition% was measured in comparison with the control group (the group treated with DMSO used as a solvent of the compound without treating the compound). For example, relative inhibition of 40% means that the binding of PCSK9 and EGF-AB in the presence of a compound is 60% relative to the control, ie, the binding rate is reduced by 40%. The results are shown in Table 1.

1.3. Experiments on the expression levels of PCSK9 and LDLR

To determine whether the 100 compounds selected in Example 1.1 affected the expression of PCSK9 and / or LDLR in cells, the expression levels of PCSK9 and LDLR in cells were measured using HepG2 cell line (ATCC: HB-8065) . First, HepG2 cells were inoculated into 2 × 10 ⁵ cells per well of a 12-well plate in medium A (100 μl / mL penicillin, 100 μg / mL streptomycin sulfate, DMEM) supplemented with 10% fetal bovine serum Lt; / RTI > After 16 hours, the cells were washed twice with PBS (phosphate buffered saline), exchanged with medium A supplemented with 10% delipidated serum (DLPS), and each compound to be tested was added at a concentration of 10 ug / mL And then cultured with the cells for 24 hours. The cultured cells were washed twice with PBS and then recovered for immunoblot analysis. The recovered cells were washed with NUN buffer (0.33 M NaCl, 1.1 M urea, 1% Nonidet P-40, 25 mM HEPES (pH 7.6), proteinase inhibitors (1 mM dithiothreitol, 10 μg / ml leupeptin, 5 μg / ml pepstatin A, 1 mM PMSF , 2 [mu] g / ml aprotinin, and 50 [mu] g / ml ALLN). After quantifying the amount of protein in the solution in which the cells were dissolved, the buffer solution for Laemmli sample was treated, heated at 100 ° C for 5 minutes, and 25 ug of protein was electrophoresed using 8% SDS-polyacrylamide gel. lt; / RTI > membrane. The membranes were incubated in blocking buffer A (1 × PBS, 0.1% Tween 20, 5% nonfat milk) for 1 hour at room temperature, and then incubated with LDLR antibody, a polyclonal antibody diluted 1: 1,000 PCSK9 antibody, respectively, and reacted at room temperature for 1 hour. The membrane was washed once with 1 × PBS containing 0.1% Tween 20 for 15 minutes, washed twice with 5 minutes to remove unbound antibody, and washed with 1: 52,000 diluted horseradish peroxidase-linked After re-reacting in blocking buffer A containing donkey anti-rabbit IgG (Pierce, Rockford, IL USA), unbound antibody was removed by washing again. The washed membrane was developed using an ECL solution. An antibody against GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as a default. The intensity of each band corresponding to PCSK9 and LDLR on the developed film was measured using ImageJ, and the amount of each protein expressed was multiplied by the amount of increased protein as compared with the group treated with DMSO as the control group. That is, when the expression level is 1.3, it means that the expression level of the protein is increased 1.3 times as compared with the control. The results are shown in Table 1.

1.4. Experiment to measure LDL-cholesterol absorption

(ATCC: HB-8065) was used to determine whether 100 compounds selected in Example 1.1 affected the absorption of low density lipoprotein (LDL) -cholesterol in cells The amount of LDL in the cells was measured. First, HepG2 cells were inoculated into 2 × 10 ⁵ cells per well of a 12-well plate in medium A (100 μl / mL penicillin, 100 μg / mL streptomycin sulfate, DMEM) supplemented with 10% fetal bovine serum Lt; / RTI > After 16 hours, the cells were washed twice with PBS (phosphate buffered saline) and replaced with medium A supplemented with 10% delipidated serum (DLPS). Each compound was added at 10 ug / mL, Lt; / RTI > The cultured cells were washed twice with PBS and then incubated with fluorescence-labeled Dil-LDL (Biomedical Technology Inc.) for 2 hours and then observed with a fluorescence microscope. Cells after the completion of the observation were collected by treatment with trypsine-EDTA, and then the amount of intracellular light was measured using a FACScalibur flow cytometer, and compared with the control group (an untreated group), the LDL absorption rate (Dil -LDL uptake) is represented by a fold value. For example, if the Dil-LDL uptake value of the compound was 1.3, the fluorescence level (intensity of fluorescence) was 1.3 times higher than that of the control group treated with DMSO, indicating that the degree of absorption of Dil- Which is 1.3 times larger than that of the previous study. The results are shown in Table 1. Lab No. is a number arbitrarily assigned to the verification process, and ChemBridge ID is the ID of the compound provided by ChemBridge's website.

Lab No. ^* Docking score ChemBridge ID Molecular Weight Relative Inhibition Language-LDL Uptake LDLR expression level PCSK9 expression level CB_1 28.40 5323858 348.5 36.8% 1.55 1.72 0.97 CB_2 27.26 5422509 419.5 49.7% 1.97 1.58 0.97 CB_3 27.12 5423970 389.5 43.9% 1.85 1.42 0.53 CB_4 28.18 5428460 400.5 31.7% 1.35 0.93 0.60 CB_5 27.15 5573554 276.3 10.7% 1.08 1.03 0.29 CB_6 27.20 5646807 393.5 17.2% 0.85 0.91 0.68 CB_7 27.00 5679233 405.5 12.6% 1.11 0.77 0.75 CB_8 28.45 5781565 415 37.3% 1.49 1.40 1.07 CB_9 28.82 5834186 445.5 12.9% 0.90 0.84 0.57 CB_10 27.78 5851930 404.5 9.9% 0.85 0.93 0.76 CB_11 27.27 5862390 397.3 4.5% 0.94 0.81 0.49 CB_12 28.36 5865258 430.2 -2.0% 1.08 0.74 0.68 CB_13 28.17 6577494 455.6 41.4% 1.66 1.77 1.54 CB_14 27.13 6578587 442.6 46.6% 2.26 1.20 1.90 CB_15 27.10 6579857 379.5 22.9% 1.41 1.44 1.07 CB_16 27.37 7000500 388.5 14.1% 1.01 0.26 0.01 CB_17 28.26 7011653 327.5 45.0% 1.56 0.92 1.11 CB_18 27.90 7017850 299.4 49.9% 1.56 0.69 0.98 CB_19 27.50 7021996 341.5 38.3% 1.59 1.37 1.20 CB_20 29.19 7300311 380.5 -1.6% 1.05 0.87 0.86 CB_21 28.47 7319736 355.9 47.5% 1.36 1.09 0.72 CB_22 27.42 7497360 449.6 11.6% 0.97 1.19 1.23 CB_23 27.95 7596514 463.6 11.7% 0.87 1.15 0.99 CB_24 27.52 7597336 422.6 12.9% 0.80 0.80 0.75 CB_25 27.75 7682179 484 12.1% 1.06 1.18 0.66 CB_26 27.48 7732594 385.4 53.4% 0.94 0.56 0.59 CB_27 27.31 7736937 376.4 14.4% 0.95 0.88 0.86 CB_28 28.26 7796312 416.5 -4.5% 0.90 0.79 0.98 CB_29 27.12 7877851 416.5 3.3% 0.76 1.32 0.74 CB_30 27.83 7883245 408.3 20.2% 0.95 1.10 1.34 CB_31 27.05 7891362 354.4 13.0% 0.87 0.87 0.91 CB_32 27.04 7922733 385.8 19.0% 0.81 0.64 0.20 CB_33 28.85 7925242 341.5 52.7% 1.85 1.86 1.80 CB_34 27.05 7925467 382.4 -2.5% 0.73 0.87 0.52 CB_35 27.18 7926470 386.9 -12.0% 0.99 0.87 0.10 CB_36 29.63 7926604 371.5 49.3% 1.69 1.57 1.22 CB_37 27.06 7941561 416.6 11.3% 0.99 0.29 0.29 CB_38 28.99 7949973 437.5 -3.2% 0.95 1.37 0.40 CB_39 27.87 7962923 432.9 10.0% 1.05 0.81 1.01 CB_40 29.17 7966968 383.5 39.9% 1.57 1.08 1.00 CB_41 27.18 7968349 372.5 12.6% 1.09 0.76 0.55 CB_42 28.00 7969162 453.5 33.6% 1.07 0.93 0.81 CB_43 27.40 7983219 430.9 31.4% 1.49 1.04 0.80 CB_44 27.58 7984456 405.9 18.2% 1.46 1.43 1.15 CB_45 28.76 7990813 440.5 -6.5% 1.08 0.83 0.98 CB_46 27.70 7998682 384.4 30.9% 1.15 0.92 0.76 CB_47 27.38 9024196 384.5 22.9% 1.31 0.99 0.68 CB_48 27.11 9025824 346.5 25.0% 1.09 1.08 0.67 CB_49 27.63 9070071 371.5 30.5% 1.39 1.43 1.64 CB_50 27.47 9148410 415.5 38.7% 1.39 1.17 1.15 CB_51 26.99 7127155 468.5 -2.6% 0.95 0.74 0.85 CB_52 26.99 7010655 295.4 15.4% 1.80 1.41 0.60 CB_53 26.98 6707210 409.6 28.4% 1.74 1.16 0.75 CB_54 26.98 7980928 391.9 2.6% 1.74 1.09 0.87 CB_55 26.97 7059604 448.4 -5.3% 1.03 0.51 0.67 CB_56 26.95 9006787 370.4 26.1% 1.02 1.40 0.50 CB_57 26.94 6991769 318.5 12.5% 2.18 1.10 0.46 CB_58 26.89 7116267 469.6 -27.9% 1.49 1.43 0.23 CB_59 26.86 6587027 339.8 -5.8% 1.18 0.53 0.00 CB_60 26.85 5537034 332.4 4.6% 0.94 0.61 0.16 CB_61 26.84 6578162 459.6 14.9% 1.03 1.26 0.74 CB_62 26.81 9011745 356.4 4.7% 1.02 0.70 0.15 CB_63 26.81 7913488 402.5 -11.1% 0.90 0.45 0.57 CB_64 26.79 6447900 400.5 -0.6% 2.04 1.27 0.72 CB_65 26.79 7261073 353.5 -1.8% 1.17 0.98 0.26 CB_66 26.78 7943026 493.4 10.2% 1.93 0.83 0.60 CB_67 26.75 5252938 378.4 -10.6% 1.08 0.44 0.79 CB_68 26.74 5426872 389.5 10.0% 0.52 0.65 0.41 CB_69 26.74 7970741 345.5 5.8% 1.22 1.18 0.88 CB_70 26.73 9025813 348.4 9.8% 2.06 1.23 0.59 CB_71 26.69 7547620 361.4 -8.5% 1.01 0.68 0.41 CB_72 26.69 5723280 330.4 25.8% 1.21 1.06 1.28 CB_73 26.68 5425499 345.5 18.9% 1.93 1.76 1.26 CB_74 26.67 7968546 341.5 19.6% 1.94 1.48 1.49 CB_75 26.67 5665101 459.5 -1.5% 1.23 0.52 0.75 CB_76 26.66 7873145 381.5 7.0% 1.10 0.70 0.33 CB_77 26.66 9036091 337.4 16.0% 1.18 0.82 1.06 CB_78 26.64 7543524 368.4 -2.7% 1.44 0.77 0.23 CB_79 26.62 5427177 398.5 52.3% 1.76 0.98 0.62 CB_80 26.62 7924529 299.4 6.9% 1.49 1.36 1.18 CB_81 26.62 7232922 431.5 -10.0% 1.35 1.20 0.15 CB_82 26.61 6759201 397.6 12.1% 1.32 1.65 1.45 CB_83 26.61 9014232 327.8 22.5% 2.08 0.97 0.61 CB_84 26.59 7997500 377.5 11.9% 1.34 1.62 1.80 CB_85 26.59 7924880 329.5 18.6% 1.44 1.31 1.22 CB_86 26.58 9038237 348.4 8.4% 1.54 1.22 0.89 CB_87 26.55 7966644 385.5 11.2% 1.80 1.60 1.37 CB_88 26.48 7595292 373.9 3.3% 1.09 0.65 0.08 CB_89 26.48 7968661 475.5 3.2% 1.26 1.21 0.39 CB_90 26.47 7839314 389.4 6.2% 0.76 1.47 0.95 CB_91 26.44 7889540 366.4 2.5% 1.12 2.08 0.76 CB_92 26.43 9030034 422.4 1.1% 1.46 1.44 0.59 CB_93 26.41 6484579 375.5 2.3% 1.18 1.26 0.78 CB_94 26.39 7280568 366.5 -5.6% 1.11 1.03 0.91 CB_95 26.35 7934110 416.5 -12.5% 0.91 1.19 0.64 CB_96 26.34 6659178 403.4 -11.3% 1.01 0.59 0.86 CB_97 26.33 9019728 371.5 -0.2% 1.44 0.97 1.01 CB_98 26.32 7633305 397.4 6.0% 1.05 0.97 0.05 CB_99 26.31 7791626 434.6 11.5% 0.91 0.74 0.84 CB_100 26.24 7933432 444.5 -3.0% 0.85 0.62 0.68

1.5. Comparison of coupling values and experimental results

SPSS statistics software (version 20) was used to confirm the relationship between the docking score (Example 1.1) measured using the GOLD program and the effect measured in vitro (Examples 1.2 to 1.4) Spearman's rank order correlation coefficient method was used. The results are shown in Table 2.

Parameter Combined value Binding inhibition rate LDL absorption LDLR expression level PCSK9 expression level Combined value (docking score) 1,000 0.444 -0.049 0.008 0.192 Relative inhibition 0.444 1,000 0.400 0.334 0.409 LDL absorption Language -LDL uptake) -0.049 0.400 1,000 0.516 0.313 LDLR expression level (amount of LDLR) 0.008 0.334 0.516 1,000 0.478 PCSK9 expression level (amount of PCSK9) 0.192 0.409 0.313 0.478 1,000

As shown in Table 2, the association values were highly correlated with the binding inhibition rate (ρ = 0.444, P <0.01), while the correlation between LDL absorption and LDLR uptake was low. (P <0.01), LDLR expression (ρ = 0.334, P <0.01), and PCSK9 expression level (ρ = 0.409, P <0.01) in the case of the binding inhibition rate Respectively. LDL absorption was correlated with LDLR uptake (ρ = 0.516, P <0.01). The above results indicate that the combination of PCSK9 and LDLR can not effectively identify compounds capable of effectively inhibiting PCSK9-LDLR binding alone, but it is also possible that the amount of PCSK9 or LDLR expressed by immunoblot, LDL absorption, LDLR binding inhibitor (PCSK9-LDLR binding inhibitor) can be selected by the addition of additional tests such as inhibition of LDLR binding.

Example 2: Confirmation of dose-dependent inhibition of PCSK9-LDLR binding of compounds

N-dimethyl-N '- (2- {2- [4- (1-methyl-1-phenylethyl) phenoxy] ethoxy} ethyl) -1,2 N-dimethyl-N '- (2- {2- [4- (1-methyl-1-phenylethyl) phenoxy] ethoxy} -ethanediamine hydrochloride (CB_36, ChemBridge ID: # 7926604, ethyl) -1,2-ethanediamine hydrochloride and a structural analogue (3D analog) of 1- {2- [4- (1-methyl-1-phenylethyl) phenoxy] ethyl} piperazine oxalate , ChemBridge ID: # 7632817, 1- {2- [4- (1-methyl-1-phenylethyl) phenoxy] ethyl} piperazine oxalate and NN- ) Phenoxy] decanamine oxalate (Chemical Formula 4), ChemBridge ID: # 7338220, N, N-dimethyl- 2- 4- (1-methyl- 1- phenylethyl) phenoxy] ethanamine oxalate. -LDLR binding.

(2)

(3)

[Chemical Formula 4]

The inhibition of PCSK9-LDLR binding was confirmed by measuring LDLR and PCSK9 expression levels and dil-LDL absorption rate in the same manner as in Example 1. The results are shown in Fig.

As shown in Figure 1, all three compounds were found to increase LDLR and PCSK9 expression levels depending on dose (Figure 1A). In the case of # 7632817, it was confirmed that LDLR and PCSK9 expression levels were decreased at 20 ug / mL, but it is presumed that the cells were killed due to cytotoxicity rather than the reduction effect by the compound (FIG. 1A) . In the case of # 7926604, the dilution rate of dil-LDL was 1.69-fold higher than that of the control group, and in the case of # 7632817, it was 2.13-fold higher when treated with the concentration of 10 ug / mL, In the case of # 7338220, it was 1.37 times as much as that of the case of treatment at a concentration of 30 ug / mL, confirming the lowest absorption rate. These results show that both CB_36 and two similar structural compounds can increase LDL-cholesterol uptake in HepG2 cell line. Thus, all of the three compounds inhibit the blood cholesterol concentration as a PCSK9-LDLR binding inhibitor It is possible to confirm that it is usable. Also, it was confirmed from the above results that the compounds selected by the method used by the present inventors can be used as a PCSK9-LDLR binding inhibitor which can be used for the prevention and treatment of cholesterol-related diseases.

Example 3: Additional screening of PCSK9-LDLR binding inhibitors

Experiments were carried out in the same manner as in Example 2 to confirm the ability of the remaining compounds to inhibit PCSK9-LDLR binding, except for compounds showing cytotoxicity. As a result, it was found that CB_1 (# 5323858); 2- {4- [4- (9H-carbazol-9-yl) -2-butyn- 2- (4- (benzyloxy) -3-methoxybenzyl] -6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline), CB3 (# 5423970; - [4- (benzyloxy) benzyl] -6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline), CB_8 (# 5781565; 4- (benzyloxy) -3-methoxybenzyl] -N- [2- (diethylamino) ethyl] -4-chlorophenyl) -1-adamantyl] ethyl} -4-piperidinamine, CB-13 (# 6577494; ] -4-piperidinecarboxamide), CB_14 (# 6578587, N-benzyl-1- [3- (benzyloxy) benzyl] -N-ethyl-4-piperidinecarboxamide oxalate), CB_15 (# 6579857 (3-methoxypropyl) {3- [4- (1-methyl-1-methylpropyl) piperidine] phenylethyl) phenoxy] propyl} amine oxalate), CB - 33 (# 7925242; N, N-dimethyl-N ' 1,2-ethanediamine hydrochloride), CB4 N- (3-methoxy-4 - [(2-methylbenzyl) oxy] benzyl} -2- (4-morpholinyl) ethanamine dihydrochloride), CB-50 (# 9148410; [Chemical Formula 18]; 2- {4- (2-methyl-3,5-diphenylpyrazolo [1,5-a] pyrimidin-7-yl) -1-piperazinyl] ethanol, CB_52 (# 7010655; 1 - [(4-biphenylyloxy) acetyl] -2-propen-1-amine oxalate), CB_53 (# 6707210) ] -4-piperidinyl} -4-ethylpiperazine hydrochloride), CB57 (# 6991769; N, N-dimethyl-N ', N'- 1,2-ethanediamine hydrochloride, CB-73 (# 5425499), 2- [4- (1-pyrenylmethyl) -1-piperazinyl] ethanol, CB-74 (# 4-biphenylyloxy) propyl] [2- (4-morpholinyl) ethyl] amine hydrochloride), CB-83 (# 9014232) ), CB - 84 (# 7997500), 2- (2-methoxy-4 - {[(2- phenylethyl) amino] methyl} phenoxy) -1-phenylethanol hydrochloride, CB - 85 N '- {2- [2- (2-biphenylyloxy) ethoxy] ethyl} -N, N-dimethyl-1,2-ethanediamine hydrochloride), and CB-87 (Formula 79) - (benzyloxy) phenoxy] butyl} It has been confirmed that the compound [2- (4-morpholinyl) ethyl] amine hydrochloride inhibits PCSK9-LDLR binding, increases LDLR and PCSK9 expression levels, and increases LDL-cholesterol absorption in HepG2 cell line.

From these results, it can be concluded that a total of 23 compounds out of the 100 compounds selected by predicting the binding level inhibit (i) binding of PCSK9 to LDLR, (ii) increase the uptake of Dil-LDL in HepG2 cells At the same time, (iii) the expression level of LDLR was increased due to the inhibition of the binding of PCSK9 and LDLR. Thus, the above-mentioned compounds were found to exhibit low cytotoxicity as a PCSK9-LDLR binding inhibitor and to decrease blood cholesterol concentration I can confirm that it is possible.

Example  4. Cholesterol concentration in mouse by compound Reduction ability

In vivo experiments were carried out using CB_36 selected in Example 1 to confirm that it reduced blood cholesterol concentrations not only in vitro but also in vivo. For in vivo experiments, 8 male male C57B / 6J mice and Pcsk9 knockout (Pcsk9 ^{- / -} ) mice were purchased from Jackson Laboratory. CB_36 compounds stored in DMSO at 20 mg / mL were diluted with PBS And was administered to the tail vein for 2 days at a concentration of 1 mg / kg / day. After 18 hours of the second administration, the mice used in the experiment were euthanized and the blood collected from the inferior vena cava was added with 2 mM EDTA and aprotinin and centrifuged to separate the plasma. The liver of each mouse was kept at -70 ° C for immunoblot analysis before the experiment. The concentration of triacylglycerol (TG) and concentration of total cholesterol (TC) in the obtained plasma were measured using Infinity ™ Triglycerides and Infinity ™ Cholesterol reagent from Thermo Scientific Inc. The concentration of cholesterol was measured using FPLC (fast performance liquid chromatography). The amount of LDLR and PCSK9 expression was analyzed by immunoblot analysis in the same manner as in Example 1.3 using liver tissues collected. The results are shown in Table 3 and FIG.

Parameter Wild type Pcsk9 ^{- / -} Control CB_36 Control CB_36 Number of mice 6 6 5 5 Body weight (g) 26.5 ± 1.2 26.5 ± 0.3 26.1 ± 0.9 27.4 ± 1.4 Liver weight (g) 1.27 + - 0.12 1.24 ± 0.03 1.23 + - 0.11 1.31 + 0.15 Liver weight / Body weight (%) 4.76 0.31 4.68 ± 0.04 4.72 ± 0.40 4.78 ± 0.34 Triglycerides (mg / dl) 77 ± 4 63 ± 5 38 ± 11 34 ± 3 Cholesterol (mg / dl) 71 ± 9 58 ± 10 58 ± 3 68 ± 11

As shown in Table 3, CB_36 decreased the total cholesterol level by 18% (P <0.05) in the wild type, but did not show a significant difference in the Pcsk9 knockout mice. From the above results, it was confirmed that CB_36 inhibits the binding of PCSK9 and LDLR, effectively lowering blood cholesterol concentration in vivo.

In addition, as shown in FIG. 2A, CB_36 decreased the high density lipoprotein, which is the 23rd to 30th sample, as well as the low density lipoprotein, which is the 15th to 22nd sample of FPLC in wild type mice Finally, it was confirmed that plasma total cholesterol levels were decreased, and that cholesterol levels were not affected in Pcsk9 - / - mice. As shown in FIG. 2B, it was confirmed that the expression level of LDLR and Pcsk9 did not change in liver tissues by CB_36. These results indicate that CB_36 decreases total cholesterol levels in plasma but does not affect the expression levels of LDLR and PCSK9 in the liver. Thus, the inhibitor of PCSK9-LDLR binding inhibits LDLR and / or PCSK9 in vivo And the total cholesterol concentration in the plasma can be effectively reduced.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (49)

A composition for preventing and treating cholesterol-related diseases, comprising a compound represented by the following formula (1) as an active ingredient:
[Chemical Formula 1]

R is _{N (CH 3) 2, N} (CH 2) 4 NH, CH 2 NH (CH 2) 3 OCH 3, CH 2 NH (CH 2) 2 N (CH 3) 2, NH (CH 2) 3 or O (CH ₂ ) ₂ NH (CH ₂ ) ₂ N (CH ₃ ) ₂ . The method according to claim 1,
Wherein said compound is at least one of the compounds represented by the following formulas (2) to (7).
(2)

(3)

[Chemical Formula 4]

[Chemical Formula 5]

[Chemical Formula 6]

(7)

The method according to claim 1,
Wherein said cholesterol-related disorder is a metabolic disease or dyslipidemia disease. The method of claim 3,
Wherein said metabolic disorder is obesity, diabetes, hypertension, or cardiovascular disease. The method of claim 3,
Wherein the dyslipidemia is hyperlipidemia, hypertriglyceridemia, or hypercholesterolemia, wherein the total cholesterol, LDL-cholesterol, or triglyceride is increased in the blood. A composition for inhibiting PCSK9 and LDLR binding comprising a compound represented by the following formula (1) as an active ingredient:
[Chemical Formula 1]

R is _{N (CH 3) 2, N} (CH 2) 4 NH, CH 2 NH (CH 2) 3 OCH 3, CH 2 NH (CH 2) 2 N (CH 3) 2, NH (CH 2) 3 or O (CH ₂ ) ₂ NH (CH ₂ ) ₂ N (CH ₃ ) ₂ . The method according to claim 6,
Wherein said compound is at least one of the compounds represented by the following formulas (2) to (7).
(2)

(3)

[Chemical Formula 4]

[Chemical Formula 5]

[Chemical Formula 6]

(7)

A composition for preventing and treating cholesterol-related diseases, comprising as an active ingredient, a statin compound and a compound represented by the following formula (1)
[Chemical Formula 1]

R is _{N (CH 3) 2, N} (CH 2) 4 NH, CH 2 NH (CH 2) 3 OCH 3, CH 2 NH (CH 2) 2 N (CH 3) 2, NH (CH 2) 3 or O (CH ₂ ) ₂ NH (CH ₂ ) ₂ N (CH ₃ ) ₂ . 9. The method of claim 8,
Wherein the compound represented by the formula (1) is any one or more of the compounds represented by the following formulas (2) to (7).
(2)

(3)

[Chemical Formula 4]

[Chemical Formula 5]

[Chemical Formula 6]

(7)

9. The method of claim 8,
Wherein said cholesterol-related disorder is a metabolic disease or dyslipidemia disease. 11. The method of claim 10,
Wherein said metabolic disorder is obesity, diabetes, hypertension, or cardiovascular disease. 11. The method of claim 10,
Wherein the dyslipidemia is hyperlipidemia, hypertriglyceridemia, or hypercholesterolemia, wherein the total cholesterol, LDL-cholesterol, or triglyceride is increased in the blood. 9. The method of claim 8,
The statin compound may be at least one selected from the group consisting of lovastatin, pravastatin, simvastatin, fluvastatin, atorvastatin, cerivastatin, rosuvastatin, &Lt; / RTI &gt; salt. A composition for preventing and treating cholesterol-related diseases, comprising a compound represented by the following formula (8) as an active ingredient:
[Chemical Formula 8]

R ₁ is any one of H, Cl, or CH ₃ , R ₂ is H, OCH ₃ Or _{O (CH 2) 4 NH (} CH 2) 2 N (CH 2) 4 O either and, R ₃ is _{H, CH 2 N (CH 2} ) 4 CHCON (CH 2) 2 CH 3 C (CH) of _{5, CH 2 NHCH 2 C (} CH) 3 O , or _{CH 2 N (CH 2) 4} CHCON (CH 2) and any one of _4, R ₄ is _{CH 2 N (CH 2) 3} C 2 (CH) 2 C _{2 (OCH 3) 2, CH} 2 N (CH 2) 4 CHCONH (CH 2) 2 N (CH 2 CH 3) 2, or _{CH 2 NH (CH 2) 2} N (CH 2) is any one of ₄ O. 15. The method of claim 14,
Wherein said compound is at least one of the compounds represented by the following general formulas (9) to (16).
[Chemical Formula 9]

[Chemical formula 10]

(11)

[Chemical Formula 12]

[Chemical Formula 13]

[Chemical Formula 14]

[Chemical Formula 15]

[Chemical Formula 16]

15. The method of claim 14,
Wherein said cholesterol-related disorder is a metabolic disease or dyslipidemia disease. 17. The method of claim 16,
Wherein said metabolic disorder is obesity, diabetes, hypertension, or cardiovascular disease. 17. The method of claim 16,
Wherein the dyslipidemia is hyperlipidemia, hypertriglyceridemia, or hypercholesterolemia, wherein the total cholesterol, LDL-cholesterol, or triglyceride is increased in the blood. A composition for inhibiting PCSK9 and LDLR binding comprising a compound represented by the following formula (8) as an active ingredient:
[Chemical Formula 8]

R ₁ is any one of H, Cl, or CH ₃ , R ₂ is H, OCH ₃ Or _{O (CH 2) 4 NH (} CH 2) 2 N (CH 2) 4 O either and, R ₃ is _{H, CH 2 N (CH 2} ) 4 CHCON (CH 2) 2 CH 3 C (CH) of _{5, CH 2 NHCH 2 C (} CH) 3 O , or _{CH 2 N (CH 2) 4} CHCON (CH 2) and any one of _4, R ₄ is _{CH 2 N (CH 2) 3} C 2 (CH) 2 C _{2 (OCH 3) 2, CH} 2 N (CH 2) 4 CHCONH (CH 2) 2 N (CH 2 CH 3) 2, or _{CH 2 NH (CH 2) 2} N (CH 2) is any one of ₄ O. 20. The method of claim 19,
Wherein said compound is at least one of the compounds represented by the following general formulas (9) to (16).
[Chemical Formula 9]

[Chemical formula 10]

(11)

[Chemical Formula 12]

[Chemical Formula 13]

[Chemical Formula 14]

[Chemical Formula 15]

[Chemical Formula 16]

A composition for prevention and treatment of cholesterol-related diseases, comprising a statin compound and a compound represented by the following formula (8) as an active ingredient:
[Chemical Formula 8]

R ₁ is any one of H, Cl, or CH ₃ , R ₂ is H, OCH ₃ Or _{O (CH 2) 4 NH (} CH 2) 2 N (CH 2) 4 O either and, R ₃ is _{H, CH 2 N (CH 2} ) 4 CHCON (CH 2) 2 CH 3 C (CH) of _{5, CH 2 NHCH 2 C (} CH) 3 O , or _{CH 2 N (CH 2) 4} CHCON (CH 2) and any one of _4, R ₄ is _{CH 2 N (CH 2) 3} C 2 (CH) 2 C _{2 (OCH 3) 2, CH} 2 N (CH 2) 4 CHCONH (CH 2) 2 N (CH 2 CH 3) 2, or _{CH 2 NH (CH 2) 2} N (CH 2) is any one of ₄ O. 22. The method of claim 21,
Wherein the compound represented by the formula (8) is any one or more of the compounds represented by the following formulas (9) to (16).
[Chemical Formula 9]

[Chemical formula 10]

(11)

[Chemical Formula 12]

[Chemical Formula 13]

[Chemical Formula 14]

[Chemical Formula 15]

[Chemical Formula 16]

22. The method of claim 21,
Wherein said cholesterol-related disorder is a metabolic disease or dyslipidemia disease. 24. The method of claim 23,
Wherein said metabolic disorder is obesity, diabetes, hypertension, or cardiovascular disease. 24. The method of claim 23,
Wherein the dyslipidemia is hyperlipidemia, hypertriglyceridemia, or hypercholesterolemia, wherein the total cholesterol, LDL-cholesterol, or triglyceride is increased in the blood. 22. The method of claim 21,
The statin compound may be at least one selected from the group consisting of lovastatin, pravastatin, simvastatin, fluvastatin, atorvastatin, cerivastatin, rosuvastatin, &Lt; / RTI &gt; salt. A composition for preventing and treating cholesterol-related diseases, comprising a compound represented by the following formula (17) as an active ingredient:
[Chemical Formula 17]

R ₁ is H or O (CH ₂ ) ₂ O (CH ₂ ) ₂ NH (CH ₂ ) ₂ N (CH ₃ ) ₂ , R ₂ is H and R ₃ is H, OCH ₂ CON ₂ ) ₄ CHN (CH ₂ ) ₄ NCH ₂ CH ₃ or O (CH ₂ ) ₃ NH (CH ₂ ) ₂ N (CH ₂ ) ₄ O and R ₄ is H. 28. The method of claim 27,
Wherein said compound is at least one of the compounds represented by the following general formulas (19) to (21).
[Chemical Formula 19]

[Chemical Formula 20]

[Chemical Formula 21]

28. The method of claim 27,
Wherein said cholesterol-related disorder is a metabolic disease or dyslipidemia disease. 30. The method of claim 29,
Wherein said metabolic disorder is obesity, diabetes, hypertension, or cardiovascular disease. 30. The method of claim 29,
Wherein the dyslipidemia is hyperlipidemia, hypertriglyceridemia, or hypercholesterolemia, wherein the total cholesterol, LDL-cholesterol, or triglyceride is increased in the blood. A composition for inhibiting PCSK9 and LDLR binding comprising a compound represented by the following formula (17) as an active ingredient:
[Chemical Formula 17]

R ₁ is H or O (CH ₂ ) ₂ O (CH ₂ ) ₂ NH (CH ₂ ) ₂ N (CH ₃ ) ₂ , R ₂ is H and R ₃ is H, OCH ₂ CON ₂ ) ₄ CHN (CH ₂ ) ₄ NCH ₂ CH ₃ or O (CH ₂ ) ₃ NH (CH ₂ ) ₂ N (CH ₂ ) ₄ O and R ₄ is H. 33. The method of claim 32,
Wherein said compound is at least one of the compounds represented by the following general formulas (19) to (21).
[Chemical Formula 19]

[Chemical Formula 20]

[Chemical Formula 21]

A composition for preventing and treating cholesterol-related diseases, comprising a statin compound and a compound represented by the following formula (17) as an active ingredient:
[Chemical Formula 17]

R ₁ is H or O (CH ₂ ) ₂ O (CH ₂ ) ₂ NH (CH ₂ ) ₂ N (CH ₃ ) ₂ , R ₂ is H and R ₃ is H, OCH ₂ CON ₂ ) ₄ CHN (CH ₂ ) ₄ NCH ₂ CH ₃ or O (CH ₂ ) ₃ NH (CH ₂ ) ₂ N (CH ₂ ) ₄ O and R ₄ is H. 35. The method of claim 34,
Wherein the compound represented by the general formula (17) is any one or more of the compounds represented by the following general formulas (19) to (21).
[Chemical Formula 19]

[Chemical Formula 20]

[Chemical Formula 21]

35. The method of claim 34,
Wherein said cholesterol-related disorder is a metabolic disease or dyslipidemia disease. 37. The method of claim 36,
Wherein said metabolic disorder is obesity, diabetes, hypertension, or cardiovascular disease. 37. The method of claim 36,
Wherein the dyslipidemia is hyperlipidemia, hypertriglyceridemia, or hypercholesterolemia, wherein the total cholesterol, LDL-cholesterol, or triglyceride is increased in the blood. 35. The method of claim 34,
The statin compound may be at least one selected from the group consisting of lovastatin, pravastatin, simvastatin, fluvastatin, atorvastatin, cerivastatin, rosuvastatin, &Lt; / RTI &gt; salt. A composition for preventing and treating cholesterol-related diseases, which comprises as an active ingredient any one or more of compounds represented by the following formulas (18), (22), (23), (24)
[Chemical Formula 18]

[Chemical Formula 22]

(23)

&Lt; EMI ID =

(25)

(26)

41. The method of claim 40,
Wherein said cholesterol-related disorder is a metabolic disease or dyslipidemia disease. 42. The method of claim 41,
Wherein said metabolic disorder is obesity, diabetes, hypertension, or cardiovascular disease. 42. The method of claim 41,
Wherein the dyslipidemia is hyperlipidemia, hypertriglyceridemia, or hypercholesterolemia, wherein the total cholesterol, LDL-cholesterol, or triglyceride is increased in the blood. A composition for inhibiting PCSK9 and LDLR binding comprising at least one of compounds represented by the following formulas (18), (22), (23), (24), (25)
[Chemical Formula 18]

[Chemical Formula 22]

(23)

&Lt; EMI ID =

(25)

(26)

A composition for prevention and treatment of cholesterol-related diseases, which comprises, as an active ingredient, at least one compound selected from the group consisting of compounds represented by formulas (18), (22), (23), (24)
[Chemical Formula 18]

[Chemical Formula 22]

(23)

&Lt; EMI ID =

(25)

(26)

46. The method of claim 45,
Wherein said cholesterol-related disorder is a metabolic disease or dyslipidemia disease. 47. The method of claim 46,
Wherein said metabolic disorder is obesity, diabetes, hypertension, or cardiovascular disease. 47. The method of claim 46,
Wherein the dyslipidemia is hyperlipidemia, hypertriglyceridemia, or hypercholesterolemia, wherein the total cholesterol, LDL-cholesterol, or triglyceride is increased in the blood. 46. The method of claim 45,
The statin compound may be at least one selected from the group consisting of lovastatin, pravastatin, simvastatin, fluvastatin, atorvastatin, cerivastatin, rosuvastatin, &Lt; / RTI &gt; salt.

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