KR20060037066A - A composition comprising glycerol compounds for prevention and treatment of cardiovascular disease - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of heart circulatory disorders comprising a glycerol compound as an active ingredient.

Glycerol-based compounds [(R)-(+)-1-glycerol monolinoleate and (R)-(-)-1-glycerol monostearate] extracted and separated from three hundred seconds according to the present invention are Lp-PLA ₂ Because of its excellent inhibitory effect, it can be useful for the prevention and treatment of cardiovascular diseases such as hyperlipidemia, coronary heart disease, arteriosclerosis and myocardial infarction caused by Lp-PLA ₂ .

Description

A composition comprising glycerol compounds for prevention and treatment of cardiovascular disease

The present invention relates to a composition for the prevention and treatment of heart circulatory disorders comprising a glycerol compound as an active ingredient.

In recent years, mortality from coronary heart disease (CHD) has increased significantly, and atherosclerosis is one of the main causes. Atherosclerosis is an inflammatory disease caused by the accumulation of lipids and fibrous elements in the artery walls. Hypertension, smoking, obesity, and increased plasma low-density lipoprotein (LDL) are the main causes. More than 50% of patients, however, suggest that there is a new cause of atherosclerosis from the fact that atherosclerosis develops regardless of the risk factors mentioned above. West Scotland Coronary Disease Prevention Research (West of Scotland Coronary Prevention Study; WOSCPS) a survey of 580 patients with a normal name 1,160 people with coronary heart disease outcome, Lp- of patients compared with control subjects PLA ₂ (Lipoprotein-associated Phopholipase through As levels of A ₂ ) were significantly higher ( N. Engl. J. Med . 2000, 343 , 1148-1155), Lp-PLA ₂ was found to be an independent risk factor for coronary heart disease ( Expert Rev. Mol Diagn , 2002, 2 , 17-22).

Lp-PLA ₂ has a molecular weight of 45 kDa and is mainly produced by phospholiphase A _2, which is produced mainly by monocytes, macrophages, T-lymphocytes and mast cells. Secreted calcium-independent member of the A ₂ superfamily (type VII), an enzyme such as platelet-activating factor acetylhydrolase (PAF-AH, EC 3.1.1.47). Known ( Arterioscler. Thromb. Vasc. Biol . 1996, 16 , 591-595). In addition, 80% of Lp-PLA ₂ is attached to LDL, and the rest is present in HDL and very low-density lipoprotein (VLDL) ( Arterioscler. Thromb. Vasc. Biol . 1995, 15 , 1764-1773).

Accumulation of LDL, especially ox-LDL, in the arterial wall is known as the most important early stage of arteriosclerosis, and Lp-PLA ₂ is latent attached to the LDL until it is oxidized / modified and then activated and rapidly oxidized. The degradation of oxidized phospholipids at a rate produces large amounts of lysophosphatidylcholine (lyso-PC) and free oxidized fatty acids ( Biochem. J. 1999, 338 , 479-487). LDL is oxidized in the intima and serves as a substrate for Lp-PLA ₂ , the degradation products in turn promote chronic inflammation associated with the accumulation of macrophages, and macrophages also produce positive Lp-PLA ₂ . A positive feedback mechanism accelerates the development of vascular disease. While the structure of free-oxidized fatty acids produced by Lp-PLA ₂ is not clearly defined, it is not possible to define all of the biological activities, but the micromolar concentrations of fatty acids produced from ox-LDL are biologically inert. In addition, reports of the pro-inflammatory and pro-atherogenic roles of another degradation product, lyso-PC, increased exponentially in the 1990s, leading to endothelial-dependent relaxation. Induction of vascular cells and intracellular adhesion molecules, acting as chemoattractants of monocytes and T-lymphocytes, producing endothelial-derived nitric oxide Over- release, inhibition of macrophage migration, toxicity at concentrations above 30-50 micromolar, and release stimulation of arachidonic acid from endothelial cells have been reported ( Curr. Opin. Pharmacol . 2001, 1 , 121-125).

Lp-PLA ₂ is an independent risk factor for coronary artery disease in patients with hypercholesterolemia, and has been suggested as a pro-inflammatory agent and found in macrophages of atherosclerosis lesions ( Arterioscler. Thromb. Vasc. Biol . 1999, 19 , 2909-2971), in a recent study, the administration of Lp-PLA ₂ inhibitors markedly produced fat streak in the Watanabe heritable hyperlipidemic rabbit, an atherosclerotic model animal. From the reduced facts ( Il Farmaco 2001, 56 , 45-50), deactivation of Lp-PLA ₂ has attracted attention as a target of atherosclerosis prevention and treatment. GlaxoSmithKline has isolated a new class of Lp-PLA ₂ inhibitors from the fermentation broth of Pseudomonas fluorescens ( J. Antibiotics 2000, 53 , 664-669), and from their organic synthetic derivatives Lp-PLA ₂ inhibitors have been developed and SB-480848 is currently in phase II clinical trials.

On the other hand, the three hundred herb (Sarurus chinensis Baill) is a perennial herb of over three hundred, and has been used in China since ancient times as an analgesic, diuretic and anti-inflammatory medicine. Since it was found in Korea that three hundred seconds lived on Jeju Island about ten years ago, it has been used in the private sector for the treatment of chronic adult diseases such as cancer treatment supplements, circulatory diseases, nervous system diseases, diabetes, etc.

The inventors of the present invention, while searching for active substances in natural resources to develop new hyperlipidemia and arteriosclerosis treatment agents with fewer side effects, were isolated from (R)-(+)-1-glycerol monolinoleate and (R) The present invention was completed by confirming the effect of inhibiting the Lp-PLA ₂ enzyme in-(-)-1-glycerol monostearate compounds.

The present invention is to provide a composition for the prevention and treatment of heart circulatory diseases comprising a glycerol compound as an active ingredient.

The present invention provides a composition for the prevention and treatment of heart circulatory disorders comprising the glycerol compound as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a cardiovascular system comprising (R)-(+)-1-glycerol monolinoleate represented by the following formula (1) or (R)-(-)-1-glycerol monostearate represented by the formula (2) as an active ingredient. It provides a composition for the prevention and treatment of diseases.

The glycerol-based compound represented by Formula 1 or Formula 2 of the present invention may be used in the form of a pharmaceutically acceptable salt, and includes all salts, hydrates, and solvates prepared by conventional methods.

The glycerol-based compound represented by the general formula (1) or the general formula (2) of the present invention can be obtained by any conventional method, commercially available reagents can be used, and in the present invention, extracted, separated and purified from 300 seconds.

Extraction, separation and purification method of the glycerol-based compound from the three hundred seconds according to the present invention is as follows.

The dried triticale root powder is extracted with ethyl acetate and concentrated under reduced pressure. As a result of measuring the Lp-PLA ₂ activity of the ethyl acetate extract, it was confirmed that the Lp-PLA ₂ inhibitory effect is excellent, the ethyl acetate extract is concentrated and dried under reduced pressure to obtain a dark brown oily substance.

The ethyl acetate extract obtained above was separated by silica gel column chromatography using a mixed solvent of n-hexane and ethyl acetate as a mobile phase. At this time, it is preferable to use a solvent having n-hexane: ethyl acetate = 9: 1 to 1: 9 (v / v) as the mobile phase.

The active fraction is separated again by silica gel column chromatography using a mixed solvent of chloroform and acetone as a mobile phase. At this time, it is preferable to use a solvent having chloroform: acetone = 33: 1 to 95: 5 (v / v).

The active fraction was separated and purified by C-18 reversed-phase silica gel column chromatography using 90% aqueous methanol solution as a mobile phase to obtain a pure compound.

By the same method as described above, (R)-(+)-1-glycerol monolinoleate (6.2 mg) and (R)-(-)-1-glycerol monostearate (3.7) per 1 kg of dried Triticalis roots, respectively. Mg) can be obtained.

The (R)-(+)-1-glycerol monolinoleate and the (R)-(-)-1-glycerol monostearate compounds according to the present invention have Lp- because the IC ₅₀ is 45 μM and 85 μM, respectively. It can be seen that the inhibitory activity against the PLA ₂ enzyme is excellent.

Therefore, the glycerol-based compounds according to the present invention can be usefully used for the prevention and treatment of cardiovascular diseases such as hyperlipidemia, coronary heart disease, arteriosclerosis and myocardial infarction caused by Lp-PLA ₂ .

The composition of the present invention may further contain one or more active ingredients exhibiting the same or similar functions in addition to the glycerol-based compounds.

The composition of the present invention can be used alone or in combination with methods using surgery, hormonal therapy, drug therapy and biological response modifiers for the prevention and treatment of cardiovascular diseases.

The composition of the present invention may be prepared by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients for administration. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, if necessary, as antioxidants, buffers And other conventional additives such as bacteriostatic agents can be added. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA.

The composition of the present invention may be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally or topically) according to the desired method, and the dosage is based on the weight, age, sex, health status, The range varies depending on the diet, the time of administration, the method of administration, the rate of excretion and the severity of the disease. The daily dosage of the glycerol-based compound of Formula 1 is about 0.1-100 mg / kg, preferably 0.5-10 mg / kg, and more preferably administered once to several times daily.

As a result of toxicity experiments by orally administering the glycerol-based compound of the present invention, the glycerol-based compound was found to have a 50% lethal dose (LD ₅₀ ) of at least 1,000 mg / kg or more by oral toxicity test.

The composition of the present invention can be added to health foods for the purpose of improving cardiovascular disease. When the glycerol-based compound of Formula 1 or Formula 2 according to the present invention is used as a food additive, the glycerol-based compound of Formula 1 or Formula 2 may be added as it is or may be used together with other food or food ingredients, according to a conventional method. Can be used as appropriate. The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, in the preparation of food or beverage, the glycerol compound of Formula 1 or Formula 2 according to the present invention is added in an amount of 1 to 20% by weight, preferably 5 to 10% by weight based on the raw materials. However, in the case of long-term intake for health and hygiene or health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. .

There is no particular limitation on the kind of food. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. The above-mentioned natural carbohydrates are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acids and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonic acid. Carbonating agents and the like used in beverages. In addition, the composition of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not critical but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred examples and experimental examples are presented to help understand the present invention. However, the following Examples and Experimental Examples are provided only to more easily understand the present invention, and the contents of the present invention are not limited thereto.

Example  : Separation and Purification of Glycerol Compounds from Three hundred Seconds

1. Extract from 300 seconds

After washing and drying 1kg of the three hundred seconds root purchased from Geochang, Gyeongsangnam-do, Korea, it was ground using a grinder to make a powder.

The white powder was added to 4 l of ethyl acetate and left to stand at room temperature for 3 days. Then, the ethyl acetate solution was stirred, filtered through a filter paper, and concentrated under reduced pressure. The extraction process was repeated twice, and only the liquid phase was collected and concentrated under reduced pressure to obtain an ethyl acetate extract (40 g). As a result of measuring the Lp-PLA ₂ activity of the ethyl acetate extract, it was confirmed that the Lp-PLA ₂ inhibitory effect is excellent, the ethyl acetate extract was concentrated and dried under reduced pressure to obtain a dark brown oily material (oily).

2. Separation and Purification from Ethyl Acetate Layer

Silica gel column chromatography (silica gel; Merck) was prepared by varying the ratio of n-hexane and ethyl acetate solution (n-hexane: ethyl acetate = 9: 1 to 1: 9 (v / v)) of the ethyl acetate extract obtained in step 1 above. , Art 9385, column size: Φ10 × 20 cm) was used to obtain an active fraction (3.5 g).

The active fraction was again changed to the ratio of chloroform and acetone solution (chloroform: acetone = 33: 1 to 95: 5 (v / v)), followed by silica gel column chromatography (silica gel; Merck, Art 9385, column size: Φ4. 2 x 20 cm) was used to obtain an active fraction (1.5 g).

The active fraction was subjected to C-18 reversed-phase silica gel column chromatography (Merck, Lichroprep C18, column size: Φ3.0 x 20 cm) using 90% methanol aqueous solution again as a pure phase (R)-(+)- 1-glycerol monolinoleate (6.2 mg) and (R)-(-)-1-glycerol monostearate (3.7 mg) were obtained.

3. Structure Analysis of Glycerol Compounds

The material obtained through the above example was determined using a VG high resolution GC / MS spectrometer, Election Ionization MS, Autospec-Ultima, and the molecular weight and molecular formula were determined by using a polarizer (Jasco DIP-). 181 digital polarimeter). Nuclear Magnetic Resonance (NMR) analysis (Bruker AMX 300) also provides ¹ H NMR, ¹³ C NMR, Homo-Cozy (HOMO-COSY), 1M-Detected heteronuclear Multiple-Quantum Coherence (HMQC), and Heteronuclear Multiple-Bond (HMBC). Coherence) and DEPT (Distortionless Enhancement by Polarization) spectra were obtained and molecular structure was determined.

The measurement results are as follows, and the active material isolated from the three hundred seconds according to the above embodiment has (R)-(+)-1-glycerol monolinoleate having the structure of Formula 1 and the structure of Formula 2 ( R)-(-)-1-glycerol monostearate was identified.

(R)-(+)-1-glycerol monolinoleate (Compound 1)

1) Physical property: Colorless oil

2) Linearity: [α] _D ²⁵ + 5 ° (c = 0.2, CHCl ₃ )

3) Molecular Weight: 354

4) Molecular formula: C ₂₁ H ₃₈ O ₄

5) ¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.89 (t, J = 11.0 μs, 3H, H-18 ′), 1.28 (m, 12H, H-4′-7 ′, H-15 ′, 16 '), 1.28 (m, 4H, H-3', H-17 '), 2.04 (m, 4H, H-8', H-14 '), 2.36 (t, J = 12.5 Hz, 2H, H -2 '), 2.77 (t like, J = 9.6, 9.6 ㎐, 2H, H-11'), 3.60 (dd, J = 9.6, 19.0 ㎐, Hα-3), 3.71 (dd, J = 6.5, 19.0 H, Hβ-3), 3.94 (m, 1H, H-2), 4.15 (dd, J = 10.1, 19.6 ㎐, Hα-1), 4.22 (dd, J = 7.8, 19.4 ㎐, Hβ-1), 5.37 (m, 4H, H-9 ', 10', 12 ', 13')

6) EIMS (rel. Int.) M / z [M] ⁺ = 354

(R)-(-)-1-glycerol monostearate (Compound 2)

1) Physical property: white powder

2) Luminous intensity: [α] _D ²⁵ -35 ° (c = 0.25, CHCl ₃ )

3) Molecular Weight: 358

4) Molecular formula: C ₂₁ H ₄₂ O ₄

5) ¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.88 (t, J = 10.5 Hz, 3H, H-18 '), 1.26 (m, 26H, H-4' to H-16 '), 1.63 ( m, 4H, H-3 ', 17'), 2.35 (t, J = 12.4 Hz, 2H, H-2 '), 3.60 (dd, J = 9.6, 19.1 Hz, 1H, Hα-3), 3.70 ( dd, J = 6.5, 19.0 μs, 1H, Hβ-3), 3.94 (m, 2H, H-2), 4.15 (dd, J = 10.0, 19.4 μs, 1H, Hα-1), 4.22 (dd, J = 7.9, 19.5 μs, 1H, Hβ-1).

6) EIMS (rel. Int.) M / z [M] ⁺ = 358

Experimental Example 1  : Lp-PLA of Glycerol Compounds According to the Present Invention ₂  Effect on activity

In order to determine the effect of the glycerol-based compound according to the invention on the Lp-PLA ₂ activity, the following experiment was performed.

1. Preparation of Enzyme Source

Blood from healthy humans was donated using a centrifuge (Sorvall Instruments RC5C) and centrifuged at 3,000 rpm and 4 ° C for 15 minutes to separate plasma and erythrocytes. To prevent denaturation of lipoproteins in this plasma, 0.04% EDTA, 0.05% NaN ₃ , 0.015% PMSF (phenylmethylsulfonyl fluoride) was added and 100,000 xg (55.2 Ti rotor, 45,000 rpm) using an ultracentrifuge (Beckman LB-70M Ultracentrifuge). ), Ultracentrifuged at 4 ° C. for 20 hours. The floating microlayer (chylomicron) and VLDL floating in the upper layer was separated, and the remaining lower layer was adjusted to 1.063 g / ㎖ by the following equation (1) using a heavy density solution (NaBr).

V2 = V1 x (D-D1) / (D2-D)

※ V2: volume of heavy density solution,

   V1: initial volume of solution,

   D: required density (1.063), D1: initial density,

   D2: density of heavy solution (1.40)

After ultracentrifugation at 100,000 x g, 4 ° C. for 24 hours, the LDL floating on the upper layer was separated. The remaining lower layer was adjusted to 1.21 g / ml by NaBr again, followed by ultracentrifugation at 100,000 × g, 4 ° C. for 24 hours, followed by separation of HDL floating on the upper layer.

gNaBr = volume x 0.94 x (Pt-Po) / 1- (V-Pt)

※ Pt: final density, Po: initial density,

   A plasma volume of 0.94: 6% is another protein, such as albumin.

   V: partial volume of NaBr at the last density.

   P (g / ml) V

    1.055 0.244

    1.063 0.244

    1.08 0.245

    1.10 0.247

    1.12 0.249

    1.14 0.250

    1.15 0.251

    1.19 0.253

    1.21 0.254

    1.23 0.254

    1.25 0.260

    1.27 0.264

    1.37 0.273

    1.40 0.270

LDL and HDL separated through this process were dialyzed with PBS (10 mM phosphate, pH 7.4) to remove high concentrations of NaBr. After dialysis, LDL and HDL were used within 1 month while stored at 4 ℃.

2. Lp-PLA ₂  Active measurement

Some modifications of the method of Boyd et al. ( Bioorg. Med. Chem. Lett. 2000, 10 , 2557-2561) were used. Standard conditions for measuring the activity of Lp-PLA ₂ were 2.7 mM EDTA, 10 mM PBS in 200 μl of the reaction solution. (pH 7.4) and 80 μM platelet activating factor (PAF; [ ³ H] PAF, 0.05 μCi / tube). That is, the substrate was prepared by removing 10 μl [ ³ H] PAF (250 μCi, 21.50 Ci / mmole) dissolved in organic solvent and 20 μl of 12.5 μM cold-PAF completely under nitrogen gas, and then removing 10 mM containing 3.4 mM EDTA. 3.2 ml of PBS (pH 7.4) was added to prepare a micelle (micellar) (A solution).

40 μl of LDL (0.5 to 0.8 mg / ml) and 160 μl of solution (A) containing 80 μM of PAF were added and reacted at 37 ° C. for 15 minutes, followed by 600 μl of chloroform / methanol (2: 1) solution. The reaction was stopped by addition. The organic solvent layer and the water layer were separated by centrifugation at 1,500 xg for 3 minutes by Bligh and Dyer's method. 250 μl of the supernatant (water layer) was taken and 250 μl chloroform was added to repeat the above separation experiment. 100 μl of the final supernatant was added to a scintillation vial and 3 ml of scintillation cocktail (Lummagel, Lumac Co.) was added to the liquid scintillation counter (1450 Microbeta Trilux, Wallac Oy, Turku, Finland). ) Was used to determine the amount of [ ³ H] acetate produced from [ ³ H] PAF (1- 0 -hexadecyl-acetyl-3H (N) -phosphatidylcholine).

The results are shown in Table 1.

compound IC ₅₀ (μM) (R)-(+)-1-glycerol monolinoleate 45 (R)-(-)-1-glycerol monostearate 85

As shown in Table 1, the (R)-(+)-1-glycerol monolinoleate and (R)-(-)-1-glycerol monostearate compounds according to the present invention have IC _{50 values} of 45 μM and 85, respectively. Indicated by μM, it can be seen that the inhibitory activity against the Lp-PLA ₂ enzyme is excellent.

Therefore, the glycerol-based compounds according to the present invention can be usefully used for the prevention and treatment of cardiovascular diseases such as hyperlipidemia, coronary heart disease, arteriosclerosis and myocardial infarction caused by Lp-PLA ₂ .

Experimental Example 2  : Acute Toxicity in Oral Administration in Mice

In order to determine the acute toxicity of the glycerol-based compounds according to the present invention, the following experiment was performed.

12 females and 12 males (3 males and 3 females each) were used as specific pathogens free ICR mice at 4 weeks of age in an animal room of temperature 22 ± 3 ° C, humidity 55 ± 10%, and illumination 12L / 12D. Breeding in. Mice were allowed to acclimate for about a week before being used in the experiment. Feed for experimental animals (JeilJedang Co., Ltd., mice and rats) and negative water were supplied after sterilization and free ingestion.

50 mg / ml concentration of (R)-(+)-1-glycerol monolinoleate and (R)-(-)-1-glycerol monostearate prepared in the above example, each using 0.5% Tween 80 as a solvent. After preparation, the mice were orally administered 0.04 ml (100 mg / kg), 0.2 ml (500 mg / kg), and 0.4 ml (1,000 mg / kg) per 20 g of the mouse body weight. Samples were administered orally once and observed for side effects or lethality for 7 days after administration. That is, on the day of administration, changes in general symptoms and the presence or absence of dead animals were observed at least 1 hour, 4 hours, 8 hours, 12 hours after the administration, and at least once every morning and afternoon from the day after the administration.

In addition, on the 7th day of administration, animals were killed and dissected, and the internal organs were visually examined. Changes in body weight were measured at daily intervals from the day of administration to determine the weight loss of animals caused by (R)-(+)-1-glycerol monolinoleate and (R)-(-)-1-glycerol monostearate. Observed.

As a result, all mice treated with test substance showed no clinical symptoms and no dead mice, and no toxicity change was observed in weight change, blood test, blood biochemistry test and autopsy findings.

Therefore, the glycerol-based compounds according to the present invention did not show a toxic change up to 1,000 mg / kg in all mice, was determined to be a safe substance with a minimum lethal dose (LD ₅₀ ) of at least 1,000 mg / kg or more.

Examples of preparations for the compositions of the present invention are illustrated below.

Formulation Example 1  : Preparation of Pharmaceutical Formulations

1. Preparation of powder

2 g of (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate)

1 g lactose

The above ingredients were mixed and filled in airtight cloth to prepare a powder.

2. Preparation of Tablets

(R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) 100 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. Preparation of Capsule

(R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) 100 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

4. Preparation of Injection Solution

10 μg / ml of (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate)

Dilute hydrochloric acid BP until pH 3.5

Injectable sodium chloride BP up to 1 ml

Dissolve (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) in an appropriate volume of injectable sodium chloride BP, and dilute the resulting solution in pH. The pH was adjusted to 3.5 with hydrochloric acid BP and the volume adjusted with sodium chloride BP for injection and mixed well. The solution was filled into a 5 ml Type I ampoule made of clear glass, dissolved under glass, enclosed under an upper grid of air, and sterilized by autoclaving at 120 ° C. for at least 15 minutes to prepare an injection solution.

Formulation Example 2  : Manufacture of food

Foods containing (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) were prepared as follows.

1. Preparation of Cooking Seasonings

(R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) 0.2 to 10% by weight for cooking for health promotion was prepared.

2. Preparation of Tomato Ketchup and Sauce

Tomato ketchup for health promotion by adding 0.2 to 1.0% by weight of (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) to tomato ketchup or sauce Sauce was prepared.

3. Manufacturing of Flour Foods

0.1 to 5.0% by weight of (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) is added to the flour and the mixture is used to make bread and cakes. , Cookies, crackers and noodles were prepared to prepare health promoting foods.

4. Preparation of soups and gravy

(R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) Add 0.1 to 1.0% by weight to soups and gravy to improve health products and noodles Soup and gravy were prepared.

5. Preparation of Ground Beef

Health promoting ground beef was prepared by adding 10% by weight of (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) to the ground beef.

6. Manufacture of Dairy Products

0.1 to 1.0% by weight of (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) is added to the milk, and the butter and ice cream using the milk Various dairy products such as

7. Manufacture of wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh.

Black beans, black sesame seeds, and perilla were also steamed and dried by a known method, and then ground to a powder having a particle size of 60 mesh.

(R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) under reduced pressure and concentration in a vacuum concentrator, dried by spraying and drying with a hot air dryer. The powder was pulverized to a particle size of 60 mesh to obtain a dry powder.

The dry powders of the grains, seeds and glycerol compounds prepared above were formulated in the following proportions.

Cereals (30% by weight brown rice, 15% by weight barley, 20% by weight barley),

Seeds (7% by weight perilla, 8% by weight black beans, 7% by weight black sesame),

Dry powder (1% by weight) of (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate),

Ganoderma lucidum (0.5% by weight),

Sulfur (0.5 wt%)

Formulation Example 3  : Preparation of Beverages

1. Preparation of Carbonated Drinks

5-10% of sugar, 0.05-0.3% citric acid, 0.005-0.02% caramel, 0.1-1% of vitamin C are mixed, and 79-94% purified water is mixed to make syrup, and the syrup is 85-98 Sterilize at 20 ℃ for 180 seconds, mix with cooling water at a ratio of 1: 4, and inject 0.5 ~ 0.82% of carbon dioxide to give (R)-(+)-1-glycerol monolinoleate (or (R)-(- ) -1 -glycerol monostearate) was prepared.

2. Manufacture of health drinks

Substances such as liquid fructose (0.5%), oligosaccharides (2%), sugar (2%), salt (0.5%), water (75%) and (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) was homogeneously blended for instant sterilization and then packaged in small packaging containers such as glass bottles and plastic bottles to prepare healthy drinks.

3. Preparation of Vegetable Juice

Health promotion vegetable juice was prepared by adding 0.5 g of (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) to 1,000 ml of tomato or carrot juice. .

4. Preparation of Fruit Juice

Health supplement fruit juice was prepared by adding 0.1 g of (R)-(+)-1-glycerol monolinoleate (or (R)-(-)-1-glycerol monostearate) to 1,000 ml of apple or grape juice. .

Glycerol-based compounds [(R)-(+)-1-glycerol monolinoleate and (R)-(-)-1-glycerol monostearate] extracted and separated from three hundred seconds according to the present invention are Lp-PLA ₂ Because of its excellent inhibitory effect, it can be useful for the prevention and treatment of cardiovascular diseases such as hyperlipidemia, coronary heart disease, arteriosclerosis and myocardial infarction caused by Lp-PLA ₂ .

Claims (4)

A composition for the prevention and treatment of cardiovascular diseases comprising (R)-(+)-1-glycerol monolinoleate represented by the following formula (1) as an active ingredient. <Formula 1>

A composition for the prevention and treatment of cardiovascular diseases comprising (R)-(-)-1-glycerol monostearate represented by the following formula (2) as an active ingredient. <Formula 2>

The method of claim 1 or 2, wherein the glycerol-based compound is extracted, separated and purified from three hundred seconds composition for the prevention and treatment of cardiovascular disease. 4. The composition of claim 3, wherein the cardiovascular disease is selected from hyperlipidemia, coronary heart disease, arteriosclerosis, and myocardial infarction.