JP3157516B2 - Angiotensin converting enzyme inhibitor - Google Patents

Description

The present invention relates to an angiotensin converting enzyme inhibitor,
In particular, it relates to substances that are effective in treating hypertension.

(Prior art) Hypertension includes essential hypertension and secondary hypertension, and angiotensin converting enzyme (hereinafter abbreviated as ACE).
Is significantly involved in diseases such as essential hypertension and secondary hypertension such as renal hypertension or endocrine hypertension. ACE acts on Angiotensin I,
The carboxyl group-side dipeptide (His-Leu) is released to produce angiotensin II having blood pressure increasing activity. Therefore, by inhibiting this enzyme, it is possible to suppress an increase in blood pressure. In fact, inhibitors have been searched from this viewpoint, and various proline derivatives have been developed and put into practical use as pharmaceuticals. However, since they are synthetic, they are unacceptable to society except for pharmaceuticals and have poor marketability for general consumers such as health foods. Natural products have been reported to inhibit green tea, soybeans, soba, and the like. However, the ability of these natural products to inhibit ACE is much weaker than that of synthetic products, and there is a need for more potent natural inhibitors.

(Problems to be Solved by the Invention) In view of the above-mentioned situation, the present inventor searched for an ACE inhibitor in a natural herbal medicine, and as a result, found that plum bark, eye grass, flatfish, Ginger, ume, tiger cane root, mao, orange peel, Chen skin,
Uwaurushi, squash, We found that there are components that strongly inhibit ACE in betel nut, gomiko, gobiko, tachibana bark, sanshuyu, tochu, kashu crow, huolian, dragon bile, physalis, kikomi, persimmon, yawami, etc. The present invention has been completed.

Accordingly, the present invention provides an enzyme inhibitor that can be used for lowering blood pressure by inhibiting the activity of ACE that causes essential hypertension and secondary hypertension such as renal hypertension or endocrine hypertension. The purpose is to provide.

(Means for Solving the Problems) Crude drugs have been used as Chinese herbs for a long time as natural pharmacologically active substances, but there are no reports on ACE inhibition of crude drugs themselves. In addition, Kampo preparations are composed of several types of crude drugs, and various effects have been clarified. Among them, some preparations have an effect of improving hypertension, but it is not clear which crude drug constituting the preparation acts on what mechanism to exert a blood pressure lowering effect. Therefore, the present inventors have repeated studies to search for a substance exhibiting a blood pressure lowering effect by inhibiting ACE from a crude drug extract, and have found that certain crude drug extracts have an inhibitory ability, and completed.

 The present invention provides an ACE inhibitor of a crude drug component.

Crude drugs used in the present invention include: As a starting material, one or more of a mixture of betel nut, gomiko, gobiko, tachibana barki, sanshuyu, tochu, kashu crow, huo lian, dragon bile, physalis, kikomi, persimmon, yawami, etc. can be used as a starting material. .

In the present invention, water, a surfactant solution, and an organic solvent are used for extracting an active ingredient from a crude drug.

Surfactants used for active ingredient extraction include a wide variety of substances, and examples thereof include the following.

Anionic surfactants Taurocholate sodium salt, etc. Cationic surfactants, amine salts, quaternary ammonium salts, etc. Amphoteric surfactants, phosphatidylethanolamine, etc. Nonionic surfactants, Tween surfactants, Triton surfactants, etc. Organic solvents used for active ingredient extraction Contains a wide variety of objects and may be exemplified by:

Alcohols (anhydrous or water-containing) Ethanol, methanol, propanol, etc. Esters Methyl acetate, ethyl acetate, etc. Ethers diethyl ether, dipropyl ether, etc. Ketones Acetone, diethyl ketone, etc. Hydrocarbons Tetrahydrofuran, benzene, etc. These water, surface activity The agent and the organic solvent can be used alone or as a mixture of two or more. In order to perform extraction using one or more crude drugs as a starting material using the above-mentioned solvent, the crude drug is extracted as it is or a pulverized product is extracted by an ordinary method. Although the extraction conditions are not particularly limited, it is preferable to perform the extraction in the range of 0 ° C to 100 ° C for about 10 minutes to 24 hours.

The amount of water, surfactant, organic solvent, etc. used in the extraction is preferably about 1 to 50 times (weight), and preferably about 2 to 5 times, the amount of the starting material.

Next, the obtained extract is subjected to solid-liquid separation to remove solids, and then concentrated if necessary to obtain an extract.

<< Enzyme activity measurement method >> The ACE inhibitory ability is measured by adding a substrate (Hip-His-Leu) to a test sample solution and an ACE solution, shaking at 37 ° C for 1 hour, and removing the resulting dipeptide (His-Leu). -After fluorescing with phthaldialdehyde, measure the fluorescence intensity (Ex. 340nm Em. 455n
m) and the ACE enzyme activity was calculated. Specifically, an ACE solution (106 mU / 5n) was added to 50 μl of the test sample solution (aqueous solution and TCA solution).
g) 20 µm and 240 µm of a 5 mM substrate solution (pH 8.3) were added, and the reaction was carried out by a conventional method. S is the fluorescence intensity of the test sample solution, A is the fluorescence intensity when water and TCA solution are added instead of the sample, B is the fluorescence intensity when water is added instead of the enzyme, and (S−B) / ACE enzyme activity was calculated from A × 100 (%). 100% when the enzyme is not inhibited at all and 0% when completely inhibited.

Next, the effects of the present invention will be further clarified by showing examples of the present invention.

(Example) Example 1 Water and a surfactant extract of a crude drug were prepared, and the ability of a water-soluble component and a poorly water-soluble component of the crude drug to inhibit ACE was examined.

First, 5 mg of the crude drug was crushed, and water and surfactant solution 1
Extract with ml at 5 ° C overnight. ACE inhibitory ability was measured for the supernatant centrifuged at 10,000 × g, 5 ° C. for 10 minutes. A 5 mM taurocholic acid solution (hereinafter abbreviated as TCA) was used as a surfactant solution.

Table 1 shows the results of the ACE inhibition test of water and the TCA extract.
Crude drugs that are effective with the water extract, that is, containing a water-soluble inhibitory substance, were fried plum bark, crow plum, mao, uwaurushi, squash, betel nut, gomiko, gobiko, and sanshuyu. TCA extract is more effective or more effective than water extract, that is, crude drugs containing poorly water-soluble inhibitors are eyegrass, tonko, ginger, wumei, tiger cane, mako, orange peel , Chen skin,
Watermelon, They were betel nut, gomiko, gobiko, tachibana skin, sanshuyu, tochu, kashu crow, huolian, dragon bile, physalis, kikomi, persimmon, and yaw.

Example 2 Among crude drugs, water and various organic solvents (methanol, ethyl acetate, acetone, tetrahydrofuran, n-hexane, diethyl ether,
Chloroform) 2 ml, extract at 5 ° C. overnight, 0.5 μm
1 ml of the extract obtained by filtration through a filter was concentrated under reduced pressure to give TCA.
After dissolving in 1 ml, the ACE inhibitory ability was measured.

The results are shown in Table 2. Inhibition was shown not only in the water extract but also in the organic solvent extract. Among the seven organic solvents, the effects were observed in methanol, ethyl acetate, acetone, tetrahydrofuran, and diethyl ether extracts.

(Effect of the Invention) As described above, the ACE inhibitor according to the present invention exhibits a strong inhibitory effect and can be provided with a safe and highly effective antihypertensive agent by being extracted from a crude drug which is a natural product .

──────────────────────────────────────────────────続 き Continued on the front page (56) References Pharmaceutical Pharmaceutical Journal, vol. 39, no. 3, pp. 245-249, 1984, Journal of the Japanese Society of Agricultural Chemistry, vol. 57, no. 11, p1143-1146, 1983 Chem. Pharm. Bull, vol. 32, no. 9, p 3615-3819, 1984 (58) Fields investigated (Int. Cl. ⁷ , DB name) A61K 35/78 A61P 9/12 A61P 43/00 BIOSIS (DIALOG) CA (STN) MEDLINE (STN)

Claims (3)

(57) [Claims] [1] A plum bark, a tongue, a ginger, a tiger cane root, an orange bark, a skin bark, a squab, a squash, a squash, a quince, a tachibana bark, a sangyuyu, a squash, a dragon, a bonito, a kiriman, and a wicker fruit Angiotensin converting enzyme inhibitor, which is a mixture containing, as an active ingredient, one or more fractions obtained by extracting a crude drug selected from the above with water, a surfactant, or an organic solvent. 2. The organic solvent includes alcohols, esters,
The enzyme inhibitor according to claim 1, which is one or a mixture of two or more selected from the group consisting of ethers and ketones. 3. The enzyme inhibitor according to claim 1, wherein the surfactant is selected from an anionic surfactant, a cationic surfactant, an amphoteric surfactant, and a nonionic surfactant.