KR20090064823A - Composition for vasorelaxant effect comprising the extract of ulmus macrocarpa - Google Patents

Abstract

A composition for relaxing blood vessel which contains an Ulmus macrocarpa extract prevent and treat various cardiovascular diseases by inducing the relaxation of vascular smooth muscle. A pharmaceutical composition for relaxing blood vessel contains an Ulmus macrocarpa extract as an active ingredient. The composition is effective in preventing and treating hypertension, coronary artery disease, cerebrovascular disease, peripheral vascular disease, main artery disease, kidney dysfunction, cardiac insufficiency or vision disorder An organic solvent for extracting the Ulmus macrocarpa is selected from ethanol, ethanol solution, methanol, methanol solution, butanol, dichloromethane, ethyl acetate, hexane and their mixture. A health food having the effect of blood vessel relaxation contains Ulmus macrocarpa extract.

Description

Vascular relaxation composition comprising the extract of baekbaekpi {COMPOSITION FOR VASORELAXANT EFFECT COMPRISING THE EXTRACT OF ULMUS MACROCARPA}

The present invention relates to a vascular relaxation composition containing the milky skin extract.

Blood pressure refers to the pressure of blood flow through the vessels, that is, arterial pressure. For some reason, systolic blood pressure is 120 mmHg (maximum blood pressure) or more and diastolic blood pressure is 80 mmHg (lowest blood pressure) or higher.

These hypertensions are primarily associated with aberrant activity of the sympathetic nervous system, dysfunction of cell membranes, impaired excretion of water and sodium ions in the kidneys, decreased secretion of antihypertensive agents such as PGE ₂ , PGI ₂ , ANP and EDRF, or catecholamines ( It is caused by abnormality of hypertension control function such as catecholamine or renin-angiotensin. That is, abnormalities of the nervous system, endocrine system, and cellular system lead to an increase in pressure of peripheral resistance blood vessels and cause hypertension. In addition, hypertension is often caused by complications such as renal failure, arteriosclerosis, cerebral hemorrhage, stroke, blindness, heart failure and heart failure.

To date, diuretics, beta blockers, calcium channel inhibitors, angiotensin converting enzyme inhibitors, and angiotensin receptor antagonists have been developed for the treatment of hypertension. However, since hypertension has been developed as a regulator for controlling hypertension, rather than a cured treatment, there is a problem of taking a drug for a lifetime while starting hypertension. Therefore, there is an urgent need for the development of new therapeutic agents that can mitigate side effects due to tissue specificity or resistance.

On the other hand, Yubaekpi (Ul 白 macro, Ulmus macrocarpa ) is a bark peeled from the root or bark of the deciduous tree, Elmus ( Ulmus macrocarpa Hance), which belongs to the elm family, and has been used as a folk medicine for a long time in Northeast Asia including Korea. It has been medicine. Before the synonym, Yubaekpi tastes sweet and flat, and acts on the parenteral, stomach, menopause, and colonoscopy, and helps to urinate well, to swell, to stool, to remove the heat of the stomach, edema, urinate, constipation, It is reported that it can be applied to cure, carbuncle, single, mastitis, and the like.

Studies on the components of milk baekpi have been studied so far in the sterols (β-sitosterol, phytosterol, stigmasterol), hydrolyzed tannin, resin, fatty oils, and catechin and catechin complexes. Proceedings have mainly been made on procyanidins and the like.

Ulmi radicis cortex extract has been reported to enhance immunity by enhancing nitric oxide (NO) production in macrophages in mouse peritoneum ( Immunopharmacol Immunotoxicol . 1999 May; 21 (2): 295-306) It has been reported that catechin polymers, called procyanidin oligomers isolated from milk extract, inhibit periodontal disease by inhibiting bacterial collagenase (J Periodontal Res. 2003 Jun; 38 (3): 282- 289). In addition, a patent application has recently been filed for the hair promoting action (Korean Patent Application No. 2005-15941) and collagen synthesis promotion and collagenase inhibitory effect (Korean Patent Application No. 2005-16084) of Yubaekpi.

The present inventors have investigated the vascular smooth muscle relaxation effect of the extracts of various herbal medicines, including baekbaekpi, as a result, it was confirmed that the extract of baekpipi showed a strong vascular smooth muscle relaxation effect, and also shows that it has an antioxidant action and antihypertensive action It confirmed and completed this invention.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for vascular relaxation, which contains milky skin extract as an active ingredient, which is useful for preventing and treating hypertension and various cardiovascular diseases caused by inducing relaxation of vascular smooth muscle.

In addition, it is an object of the present invention to provide a health functional food having a vascular relaxation effect using the milky skin extract as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for vascular relaxation containing the milk extract extract as an active ingredient.

Since the composition containing the milk extract according to the present invention as an active ingredient has an excellent blood pressure lowering effect through the smooth muscle relaxation action of blood vessels, as well as the prevention and treatment of hypertension, complications caused by hypertension, namely stroke, arteriosclerosis, ischemic It can be usefully used for the prevention and treatment of various cardiovascular diseases such as heart disease and congestive heart failure.

The pharmaceutical composition according to the present invention is characterized in that the baekbaekpi extract exhibiting a blood pressure lowering effect through the vascular smooth muscle relaxation and antioxidant action as an active ingredient.

First, the milky skin extract according to the present invention can be prepared by solvent extraction of milky skin or dried product thereof. Specifically, 2 to 200 times, preferably 10 to 30 times the organic solvent of the milky skin volume is added to the dry and fine milky skin, and 1 to 20 days at 10 to 30 ° C, preferably 5 to 10 days. The milky skin extract can be prepared by extracting, filtering and concentrating under reduced pressure. At this time, the extraction solvent may be a solvent selected from the group consisting of ethanol, ethanol aqueous solution, methanol, methanol aqueous solution, butanol, dichloromethane, ethyl acetate, hexane and mixtures thereof, 80 to 100% ethanol or ethanol aqueous solution This is preferred.

Since the milk extract of the present invention exhibits an excellent blood pressure lowering effect through vascular smooth muscle relaxation and antioxidant action, it can be used for the prevention or treatment of hypertension or high blood pressure complications.

The "hypertension" refers to a state in which the systolic blood pressure, the diastolic blood pressure or the average arterial pressure is continuously maintained higher than normal in a stable state, and the systolic blood pressure is continuously maintained at 120 mmHg or more and the diastolic blood pressure is 80 mmHg or more. Say when it will be.

"Hypertensive complications" refers to diseases caused by an abnormally elevated state of blood pressure, such as, but not limited to, coronary artery disease, cerebrovascular disease, peripheral vascular disease, aortic disease, renal dysfunction , Heart failure and visual impairment. Examples of coronary artery disease include angina pectoris and myocardial infarction, and examples of cerebrovascular diseases include stroke, cerebral hemorrhage, and cerebral infarction.

Furthermore, the pharmaceutical compositions according to the invention can be administered individually or in combination with other types of blood pressure lowering agents.

The milk extract may be mixed as an active ingredient with a suitable pharmaceutically acceptable carrier or excipient according to a conventional method or diluted with a diluent to prepare a pharmaceutical composition having the above-mentioned function. Examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl Cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition may further include fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like.

The pharmaceutical compositions of the present invention may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. The formulations may be in the form of tablets, pills, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders and the like.

The pharmaceutical compositions of the invention can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular. Typical daily dosages of the pharmaceutical compositions of the present invention range from 1 to 100 mg / kg body weight, preferably 10 to 30 mg / kg body weight, once or several, based on the active ingredient. It can be administered in divided doses. However, the actual dosage should be determined in light of several relevant factors such as the route of administration, the age, sex and weight of the patient, and the severity of the disease, and therefore the dosage does not limit the scope of the invention in any aspect. .

In addition, the present invention provides a health functional food or beverage composition containing the milk extract, which can prevent or alleviate hypertension and hypertension complications.

Foods to which the milk extract of the present invention can be added to exhibit the above effects include, for example, various foods, beverages, snacks, confections, gums, ice creams, tea bags, instant teas, granules, flavors, and vitamin complexes. , And other health supplements, but are not limited thereto.

The milk functional extract of the present invention may be added to the raw material during food preparation or appropriately mixed with the cooked food to prepare the above functional food or beverage, in which case the content of the milk extract is finally prepared in the food or beverage. May each range from 0.01 to 30% by weight.

The pharmaceutical composition, or nutraceutical or beverage of the present invention may further include other medicinal herbs or extracts thereof which are pharmaceutically acceptable to enhance or supplement the desired effect, and representative examples of such herbal medicines are bark, rhubarb , Cows and lotus roots. The herbal medicine may be used in an amount of 0.01 to 50% by weight based on the total weight of the composition.

Hereinafter, the present invention will be described in more detail based on the following examples. However, the following examples are not intended to limit the invention only.

Example 1 Preparation of Milky Skin Extract

5L ethanol was added to 1 kg of dried milky skin and chilled for 1 week, and the filtrate and the residue were separated using a filter paper (Whatman No. 2 filter paper). An equal amount of ethanol was added to the residue, and the resultant was cooled for one week, and then the filtrate and the residue were separated using a filter paper in the same manner. The extraction and filtration were repeated one more time, and the filtrates were combined and concentrated under reduced pressure at 40 ° C. to obtain 127 g of baekbaekpi ethanol extract, which was stored at 4 ° C.

Experimental Example 1 Measurement of Antioxidant Effect of Milk Extract

The antioxidant effect of the ethanol extract of milky skin obtained in Example 1 was measured as follows.

(1) Active radical scavenging ability through DPPH (1,1-diphenyl-2-picrylhydrazyl) reaction

First, the measurement of active radical scavenging ability through the DPPH reaction, each sample was prepared by concentration, 100 μl of 0.15 mM DPPH solution was added to 100 μl of each sample, mixed for 10 seconds with a plate shaker (Lab-line instruments), and then at room temperature for 30 minutes. The reaction was induced. Then, the absorbance was measured at 520 nm using a spectrophotometer. As a positive control drug, vitamin E (α-Tocopherol) was prepared by concentration. In order to compare and examine the antioxidant capacity of each sample, EC _50, which is the amount of sample required to reach an electron donating ability (EDA) of 50%, was measured.

As a result, as shown in Figure 1, the milk extract is lower than vitamin E, but showed a significant radical scavenging ability and the antioxidant effect accordingly.

(2) LDH (lactate dehydrogenase) enzyme activity and MTT 'colorimetric analysis

Cytoprotective effect of oxidative damage of the milk extract of the present invention was determined by MTT colorimetric analysis and LDH enzyme activity analysis in cells exposed to hydrogen peroxide (H9c2 cells). First, remove the medium from the cultured cells, mix lightly with PBS, remove PBS again, incubate for 5 minutes at 37 ° C with trypsin / EDTA (0.1%, 1 mM) solution, and completely remove cells from the bottom of the dish. After observation under microscope, the culture medium was added and mixed well to adjust the cell number to 1 × 10 ⁵ cells / mL, and then inoculated into 24-well plates, in advance at 37 ° C. in a 5% CO ₂ incubator for 2 days. It was used after incubation. Then, the extract obtained from milky skin was dissolved in PEG400 and then adjusted to a concentration of 1-100 μg / mL, and pretreated 30 minutes before hydrogen peroxide treatment. Plates treated with 2 mM hydrogen peroxide were left in a CO ₂ incubator for 2 hours to induce oxidative damage, and then analyzed for LDH enzyme activity released from damaged cells from the medium. Specifically, NADH solution (0.2 mg / 2.85mL) was prepared in advance, put in a tube of 0.95 mL and stored at room temperature, and made 22.7 mM sodium pyruvate solution and refrigerated and used in the experiment. 17 μl of the medium of 2 mM hydrogen peroxide and 0.95 mL of NADH solution were mixed and reacted at room temperature for 20 minutes. Then, 33 μl of sodium pyruvate was added and mixed well. The absorbance was changed at 340 nm using a spectrophotometer. Was measured and the result is shown in FIG. In this case, the non-treated group is a normal cell line without oxidative damage not treated with hydrogen peroxide, and the control group refers to a cell line that induced oxidative damage by adding 100 μl of PEG400 instead of the sample.

From FIG. 2a, it was confirmed that LDH activity, which is an apoptosis indicator enzyme, was significantly increased according to hydrogen peroxide treatment, and that LDH activity was significantly decreased when pre-treated with milk extract according to the present invention.

Meanwhile, 30 μl of MTT solution (2 mg / mL) was added to each well, and the cells were further incubated for 1 hour so that MTT was reduced by enzymatic action of viable cells. Each experimental group used three wells under the same conditions. At the end of the culture, the cell culture solution treated with MTT solution was removed, 500 μl of dimethyl sulfoxide (DMSO) was added to the lower layer to dissolve the resulting formazan crystals, and 100 μl of each DMSO was transferred to a 96-well plate. Absorbance at 540 nm was measured with a plate reader (VICTOR II, Perkin Elmer, USA). In this case, 100 μl of PEG400 was added instead of the sample. Next, cell viability (%) ((absorbance of sample treated cells / absorbance of control group) x 100) was calculated, and the results are shown in FIG. 2B.

As shown in Figure 2b, the number of viable cells increased with the treatment of the milk extract, according to the present invention, from which the cytoprotective effect from the oxidative damage of the milk extract can be confirmed.

(3) DNA protection effect

In order to reaffirm the protective effect of the milky skin extract against oxidative damage, a comet assay by Rojas et al. , Rojas E et al. , 1999, J. Chromatogr.B. Biomed. Sci. Appl. 722 , 225-254). After mixing 0.5% low-melting agarose gel and 200-300 cells per each sample in advance, place it on a microscope slide and pre-cool the cell lysis solution (2.5 M NaCl, 100 mM EDTA, 10 mM Tris pH). 10.0, 10% DMSO, 1% Triton X-100) was immersed in a cold chamber at 4 ° C. for 3 hours to induce protein dissolution. The induced slides were electrophoresed at 30 V and 250 mA for 7 minutes, and then immersed in 1 M ammonium acetate solution (70% ethanol) and 1 mg / mL spermine (70% ethanol) solution. DNA fluorescence was induced with a solution of SABER Green (Sigma-aldrich Co.), a DNA fluorescent dye. Images collected using a fluorescence microscope were measured for DNA damage by CASP program (download from www. Casp.of.pl). At this time, 30 images per slide were collected from three slides per group, and the results are shown in FIG. 3. The non-treated group is a normal cell line without oxidative damage not treated with hydrogen peroxide, and the control group refers to a cell line inducing oxidative damage by adding 100 μl of PEG400 instead of the sample and following the same procedure.

From Figure 3, it can be seen that the milky skin extract has a protective effect from oxidative DNA damage in the cell.

Experimental Example 2 Measurement of Vascular Relaxation Effect of Milky Skin Extract

The extracted vascular relaxation effect (ie, the protective effect against vasoconstriction as a blood pressure lowering effect) of the extract of the milky skin obtained in Example 1 was measured as follows.

Sprague-Dawley (SD, male) rats (Orient, Korea) were kept at room temperature (22.5 ± 1 ℃), constant humidity (55 ± 5%), and at least 2 hours in clean animal room with automatic contrast control at 12 hour intervals. It was used for the experiment after stabilizing for more than a week. After striking and stunning the back of the SD rat, the carotid artery was cut and blood was lost. After the incision of the rib cage, the descending aorta was quickly removed and the adipose tissue and surrounding tissue were carefully removed so as not to damage the endothelium. From this, the blood vessels were cut into 2-3 mm lengths (Krebs-Henseleit or Krebs' bicarbonate buffer: NaCl 118.0 mM, KCl 4.7 mM, CaCl ₂ 2.5 mM, MgSO ₄ 1.2 mM, KH ₂ PO ₄ 1.2 It was suspended in a tissue chamber containing 25.0 mM of NaHCO ₃ and 11.0 mM of glucose, Junsei Co., Japan, hereinafter referred to as "KH buffer". At this time, the temperature in the chamber was maintained at 37 ℃, saturated with O ₂ / CO ₂ mixed gas (O ₂ : CO ₂ = 95: 5) to maintain a pH of 7.4. Equilibration was carried out for 90 minutes at 2 g of initial tension and exchanged with fresh KH buffer at 15 minute intervals. Vasoconstriction / relaxation can be achieved by using a force displacement transducer (Grass FT03, Grass Co., USA) with a chart recorder (product name: Multicorder MC 6625, Hugo Sachs Co.). , German).

To adapt the bleeding vessels in the tissue chamber, it was contracted with 0.3 μM of phenylephrine (Sigma-aldrich Co., USA) and then washed three times over 45 minutes with K-H buffer. After the blood vessel was contracted with phenylephrine again to reach a stable state, 1 μM of acetylcholine chloride (Sigma-aldrich Co., USA) was added to confirm that the blood vessels were completely relaxed. . The milky skin extract used in this experiment was dissolved in DMSO and diluted with K-H buffer solution, and the DMSO concentration at the highest concentration was 0.03%. All drugs and reagents used in the experiment were prepared just before the start of the experiment.

Vascular relaxation measurement was performed by adding a sample to the blood vessels contracted by phenylephrine by concentration accumulation method (1, 3, 10 and 30 µg / ml) to measure the degree of relaxation of blood vessels. At this time, the experimental results showed the degree of relaxation based on the contraction state by phenylephrine in percentage (%), and L-NAME, which is an endothelial vascular and nitric oxide inhibitor, was used to identify the relaxation mechanism. Vascular smooth muscle by milky skin extract from pretreated blood vessels (FIG. 4A), blood vessels contracted to 45 mM potassium concentration (FIG. 4B), and blood vessels pretreated with TEA, a calcium-activated potassium channel (K _Ca ) blocker (FIG. 4C) The relaxation effect was measured.

4a to 4c, it can be seen that the milk extract has a strong vascular relaxation effect (EC ₅₀ : 1.84 ㎍ / ㎖), this relaxation effect is the mechanism of the activity of the endothelial K _Ca channel and the increase in nitric oxide secretion It is believed to depend on.

Experimental Example 3 Determination of Antihypertensive Effect of Milk Extract

The antihypertensive effect of the baekbaekpi extract obtained in Example 1 was measured as follows.

Purchased from Spontaneously hypertensive rats (SHR, male, 13-14 weeks old, Charles River Laboratories, USA) and automatically contrasted at constant temperature (22.5 ± 1 ° C), humidity (55 ± 5%), and 12 hour intervals. It was used in the experiment after stabilization in a controlled clean animal kennel. Blood pressure and heart rate were measured using a tail-cuff method, a method of measuring blood pressure from the tail of an experimental animal, using a photoplethysmograph (Model 31, IITC Life Sci., Woodland Hills). , CA, USA). For stable blood pressure measurement, the experimental animals were stabilized in a constant temperature box at 32 ° C. for about 5 to 10 minutes, and blood pressure measurement training was performed for 1 to 2 weeks before starting the experiment. SHR used only systolic blood pressure of 170 mmHg or more.

The experimental group was treated with a control group (1% Arabian gum), milky skin ethanol extract (100 mg / kg) group, control group captopril (30 mg / kg) group, and WHR rat (Wistar-Kyoto rat; Charles River). Laboratories Japan Inc .; normal blood pressure rats; 1% Arabian gum), and divided into four groups, except for WKY rats. The number of silver was seven, and the other group was eight. Each test substance was suspended using 1% Arabian gum and then orally administered once daily using sonde. All test substances used in the experiment were prepared immediately before use. Blood pressure was measured 2.5 hours after oral administration of the test substance, and the day before the start of the experiment and 1, 2, 7, 14, 21, 28, 35 and 42 days after the test substance administration. All experimental results were expressed as mean ± standard error of measurement (SEM), and statistical analysis of the experimental results was performed using the Sigma Stat (Jandel Co., USA) program, t-tested (unpaired). And one-way analysis of variance (ANOVA), and the secondary test was a dunnett multiple comparisons test.

As a result of the experiment, the amount of blood pressure change (or the blood pressure after administration of the test substance minus the blood pressure before administration of the test substance; mmHg) after orally administering the ethanol extract of milky skin, a test substance, to the SHR is shown in FIG. 5A.

In FIG. 5A, the reference blood pressures of the control group (1% Arabian gum), the 100 mg / kg ethanol extract of milky skin and the control captopril 30 mg / kg administration group were 190 ± 5.7, 195 ± 6.9 and 189 ± 3.6, respectively. There was no significant difference between groups as mm Hg, and systolic blood pressure was 142 ± 3.6 mmHg in WKY rats. Blood pressure was measured at weekly intervals for 42 days. As time passed, systolic blood pressure of WKY rats tended to decrease gradually, and the SHR control group showed a gradually rising blood pressure. When the 100 mg / kg ethanol extract of milky skin was administered, blood pressure dropped about 20% from 7 days after the administration, which continued until 42 days after the end of the experiment. When control group captopril was administered at a high dose of 30 mg / kg, it showed a significant blood pressure lowering effect than the control group.

On the other hand, the change in heart rate after administration of the test substance (heart rate after administration of the test substance minus heart rate before administration of the test substance; heart rate / minute) is shown in Figure 5b.

In FIG. 5B, the reference heart rates in the control group, 100 mg / kg ethanol extract of milky skin and 10 mg / kg control group captopril were 424 ± 10.0, 409 ± 10.5 and 406 ± 7.8 beats / min, respectively. There was no significant difference between. At this time, compared to the control group did not show a statistically significant difference in all the administration group, in the captopril administration group the heart rate increased by about 30 to 40 beats / min due to the blood pressure drop reflex action.

Experimental Example 4 Measurement of Vascular Relaxation Effect of Milky Skin Extract

After the measurement of the antihypertensive effect performed in Experimental Example 3, the vascular relaxation effect of the thoracic aorta extracted from congenital hypertensive rats treated with milky skin extract for 42 days was measured in a similar manner to Example 3. Vascular relaxation effect was measured by adding acetylcholine and sodium nitroproxide, which are vasodilators, to the blood vessels contracted by phenylephrine by concentration accumulation method (1, 3, 10 and 30 µg / ml). The experimental results are shown in Figure 6a and 6b as a percentage (%) of the degree of relaxation based on the contraction state by phenylephrine.

6A and 6B, when looking at the vascular relaxation effect by acetylcholine showing endothelial dependent vascular relaxation effect, blood vessels extracted from the control SHR (43.3 ± 3.4%) were WKY (77.7 ± 3.0%) which is a control animal of SHR. Vascular relaxation was significantly reduced compared to the blood vessels extracted from, and the blood vessels (58.9 ± 3.4%) extracted from SHR after 42 days of oral administration of ethanol extract of Yubaekpi, a test substance, showed significant vascular relaxation effect. Moreover, the vascular relaxation effect by sodium nitroproxide, which showed endothelial independent vascular relaxation effect, was 77.1 ± 2.4% in the control group and 97.1 ± 0.9% in the WKY, a control animal of the SHR, while the ethanol extract of the test substance milky skin SHR administered orally for 42 days showed a significant vascular relaxation effect of 95.2 ± 1.4%.

Experimental Example 5 Measurement of Antioxidant Effect in Congenital Hypertension Rats Treated with Milk Extract

After the measurement of the antihypertensive effect performed in Experimental Example 3, the lipid peroxidation content in the liver extracted from congenital hypertensive rats treated with milky skin extract was measured for 42 days. The extracted liver was quickly frozen in liquid nitrogen, Tris buffer (20 mM, pH 7.2) was added to the sample at a 1: 2 w: v ratio, and then ultra-turrax (IKa® Works, Model). T75 Basic, Japan) was used to make a homogeneous suspension. After centrifugation at 4000 × g for 10 minutes at 4 ° C., the content of malondialdehyde (MDA), an indicator of lipid peroxidation content, was determined from the supernatant. Specifically, MDA content was measured according to the measurement method provided by the manufacturer (Bioxytech MDA-586, OxisResearch, USA), and the amount of protein was a Bradford method using bovine serum albumin (BSA) as a standard indicator. It was measured using, and the results are shown in FIG.

In FIG. 7, MDA content in each test subject was 29.1 ± 0.9 nM / g Liver in WKY, 45.7 ± 2.6 nM / g Liver in SHR, 37.4 ± 1.9 nM / g in SHR administered orally for 42 days with ethanol extract of milky skin. It was shown as Liver, from which the significant reduction of lipid peroxidation can be confirmed according to the treatment of the milk extract.

Using the milk extract of the present invention, a pharmaceutical formulation or food was prepared as follows.

Production Example 1 Powder

The powders were prepared by mixing the following ingredients and then filling the airtight cloth according to a conventional powder preparation method:

2 g of dried milk extract

1 g lactose

Preparation Example 2 Tablet

The tablets were prepared by mixing the following ingredients and then tableting according to the conventional tablet preparation method:

Milk White Skin Dry Extract 100mg

100 mg corn starch

Lactose 100 mg

2 mg magnesium stearate

Preparation Example 3 Capsule

The capsules were prepared by mixing the following ingredients and filling the gelatin capsules according to a conventional capsule preparation method:

Milk White Skin Dry Extract 100mg

100 mg corn starch

Lactose 100 mg

2 mg magnesium stearate

<Manufacture example 4> Wire type

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to make a powder having a particle size of 60 mesh. Black beans, black sesame seeds, and perilla were also steamed and dried in a known manner, and then ground to a powder having a particle size of 60 mesh.

The milky skin extract of the present invention was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a spray hot air dryer was pulverized with a particle size of 60 mesh using a grinder to obtain a milky skin extract dry powder.

Granules were prepared by combining the dry powders of the grains, seeds and milk extracts prepared above in the following proportions.

Cereals: Brown rice 30% by weight, barley 15% by weight, barley 20% by weight, glutinous rice 9% by weight,

Seeds: perilla 7% by weight, black beans 8% by weight, black sesame 7% by weight,

3% by weight of dried milk powder, 0.5% by weight of ganoderma lucidum, 0.5% by weight of turmeric

Preparation Example 5 Chewing Gum

Chewing gum was prepared by a conventional method by combining 20% by weight of a gum base, 76.9% by weight of sugar, 1% by weight of perfume, and 2% by weight of water and 0.1% by weight of the milky skin extract of the present invention.

Production Example 6 Candy

Candy was prepared by combining 60 wt% of sugar, 39.8 wt% of starch syrup, 0.1 wt% of fragrance, and 0.1 wt% of the milk extract of the present invention.

Production Example 7 Biscuits

Force 25.59% by weight, 22.22% by weight of gravity, 4.80% by weight, white salt 0.73% by weight, 0.78% by weight, palm shortening 11.78% by weight, ammonium 1.54% by weight, 0.17% by weight sodium bisulfite 0.16% by weight , Rice flour 1.45 wt%, Vitamin B₁0.0001 wt%, Vitamin B₂0.0001 wt%, Milk flavor 0.04 wt%, Water 20.6998 wt%, Whole milk powder 1.16 wt%, Substitute milk powder 0.29 wt%, Monobasic calcium phosphate 0.03 wt% , Biscuits were prepared in a conventional manner by combining 0.29% by weight of spraying salt and 7.27% by weight of spray oil with 1% by weight of the milk extract of the present invention.

<Manufacture example 8> Healthy drink

0.26% by weight of honey, 0.0002% by weight of thioctoamide, 0.0004% by weight of nicotinic acid, 0.0001% by weight of sodium riboflavinate, 0.0001% by weight of pyridoxine hydrochloride, 0.001% by weight of inositol, 0.002% by weight of orthoic acid and 98.7362% by weight of water 1% by weight of the extract of the milky skin extract was prepared in a conventional manner for a healthy drink.

Preparation Example 9 Health Supplement

55% by weight of spirulina, 10% by weight of guar gum enzyme digestion, 0.01% by weight of vitamin B ₁ hydrochloride, 0.01% by weight of vitamin B ₆ hydrochloride, 0.23% by weight of DL-methionine, 0.7% by weight of magnesium stearate, 22.2% by weight of lactose and 1.85% by weight of corn starch % And 10 wt% of the milk extract of the present invention were formulated to prepare a tablet-type dietary supplement in a conventional manner.

1 is a result of measuring the active radical scavenging ability through the DPPH reaction of the ethanol extract of milky skin according to the present invention,

Figure 2a is a result of LDH enzyme activity analysis of ethanol extract of milky skin according to the present invention,

Figure 2b is the MTT colorimetric analysis of ethanol extract of milky skin according to the present invention,

3 is a result of measuring the DNA protection effect of ethanol extract of milky skin according to the present invention,

Figure 4a is a result of measuring the vascular smooth muscle relaxation effect in the endothelium of the ethanol extract of milky skin according to the present invention and blood vessels pretreated with L-NAME which is an inhibitor of nitric oxide synthase.

Figure 4b is a result of measuring the vascular smooth muscle relaxation effect in the blood vessels contracted to the concentration of 45 mM potassium ethanol extract of milky skin according to the present invention,

Figure 4c is a result of measuring the vascular smooth muscle relaxation effect in the blood vessels pretreated with TEA, a calcium activated potassium channel (K _Ca ) blocker of the ethanol extract of milky skin according to the present invention,

5 is an antihypertensive effect and heart rate measurement results of the ethanol extract of milky skin according to the present invention,

Figure 6 is the result of measuring the vascular relaxation effect of the ethanol extract of milky skin according to the present invention,

7 is a result of measuring the lipid peroxidation degree of ethanol extract of milky skin according to the present invention.

Claims (5)

Pharmaceutical composition for the prevention or treatment of hypertension, coronary artery disease, cerebrovascular disease, peripheral vascular disease, aortic disease, renal dysfunction, heart failure or vision disorder, containing the extract of Ulmus macrocarpa as an active ingredient. The method of claim 1, The milky skin extract is a pharmaceutical composition, characterized in that extracted with an organic solvent selected from the group consisting of ethanol, ethanol aqueous solution, methanol, methanol aqueous solution, butanol, dichloromethane, ethyl acetate, hexane and mixtures thereof. A dietary supplement for the improvement of hypertension, coronary artery disease, cerebrovascular disease, peripheral vascular disease, aortic disease, renal dysfunction, heart failure or vision disorder, containing the extract of Ulmus macrocarpa . The method of claim 3, Health functional food, characterized in that the food is in the form of a drink. The method of claim 3, Health functional food characterized in that the extract is extracted with an organic solvent selected from the group consisting of ethanol, ethanol aqueous solution, methanol, methanol aqueous solution, butanol, dichloromethane, ethyl acetate, hexane and mixtures thereof.

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