Application of 3 ', 5-dimethyl hesperidin and pharmaceutical composition with angiotensin converting enzyme inhibition effect

Abstract

The invention relates to the technical field of biology, in particular to application of 3 ', 5-dimethyl hesperidin and a pharmaceutical composition with an angiotensin converting enzyme inhibiting effect, and 3', 5-dimethyl hesperidin and 3 '-methyl hesperidin exert the angiotensin converting enzyme inhibiting effect which is more effective than that of single use of 3', 5-dimethyl hesperidin and 3 '-methyl hesperidin in a certain proportion. The new application of the 3 ', 5-dimethyl hesperidin and 3'-methyl hesperidin composition is found for the first time, and the 3 ', 5-dimethyl hesperidin and 3'-methyl hesperidin composition can be used for preparing the medicine for reducing blood pressure.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an application of 3',5-dimethyl hesperidin and a pharmaceutical composition having angiotensin converting enzyme inhibitory effect. Background Art

[0002] Hypertension is a common chronic disease and the most important risk factor for cardiovascular and cerebrovascular diseases. At present, clinical antihypertensive drugs can be divided into five categories according to their mechanism of action, namely diuretic antihypertensive drugs, sympathetic nerve inhibitors, renin-angiotensin system inhibitors, calcium antagonists, and vasodilators. Renin-angiotensin system inhibitors can inhibit the activity of ACE, reduce the generation of ATⅡ, dilate blood vessels, relieve or reverse cardiovascular structure, and play an effective role in lowering blood pressure. Among them, angiotensin converting enzyme inhibitors and angiotensin II receptor blockers are currently the most commonly used antihypertensive drugs. Angiotensin converting enzyme (ACE) is an important enzyme that converts angiotensin I into angiotensin II. Angiotensin II can cause vasoconstriction and increase blood pressure; it promotes the release of aldosterone from the adrenal cortex, and inhibiting the activity of ACE can directly prevent hypertension.

[0003] 3'-Methyl hesperidin (CAS11013-97-1) is a synthetic product in which the 3'-phenolic hydroxyl group of the natural dihydroflavonoid compound hesperidin is methylated. It has a vitamin P-like effect and can be used in combination with vitamin C to participate in various metabolic processes in the body. In the published patents, 3'-methyl hesperidin is mostly used as a component in a mixture or composition. The combination of potassium azelayl diglycinate, water-soluble hesperidin, vitamin C and vitamin B in "A composition for treating androgenic alopecia and its shampoo" (CN106937957B) can be used to treat androgenic alopecia; the probiotic fermentation supernatant, salt-soluble hesperidin, vitamin C and vitamin B in "A polysaccharide salt spray mixture containing probiotic fermentation liquid for protecting the respiratory tract and its application" (CN115154487B) The combination of liquid, modified lentinan, methyl hesperidin and oxidized linalool can be used to protect the respiratory tract; the combination of phenols, 2,4,5,6-tetrachlorophthalonitrile, rue leaf extract, dodecylbenzyldimethylammonium salt, glycitin, phellodendron extract, methyl hesperidin and allicin in "An antibacterial filter material and its anhydrous preparation process" (CN107497396B) can be used to prepare antibacterial materials; there is currently no public research or patent showing that it has a clear inhibitory effect on angiotensin converting enzyme.

[0004] 3',5-dimethyl hesperidin is a homologue of 3'-methyl hesperidin, and the 5-position of 3'-methyl hesperidin is phenolic hydroxyl group while that of 3',5-dimethyl hesperidin is methoxyl group. However, the 5-position substituent of flavonoids has a great influence on the pharmacological activity. Public literature shows that flavonoids and flavanones have the strongest anti-inflammatory activity when there is a methoxyl group at the 5-position, while the activity is significantly reduced when there is a hydroxyl group at the 5-position (Panthong A., et al. "Antiinflammatory activity of flavonoids." Phytomedicine (1994).); the anti-tumor effect of flavonoids is affected by their structure, and the phenolic hydroxyl substitution pattern and number and hydroxymethoxy substitution all affect their anti-cancer effect (Joyeux, M., et al. "Comparative Antilipoperoxidant, Antinecrotic and Scavanging Properties of Terpenes and Biflavones from Ginkgo and some Flavonoids." Planta Medica (1995).) There is little research on its pharmacological effects by scholars at home and abroad. There is no public research or patent showing that it can lower blood pressure, nor is it clear what pharmacological effects it has. Summary of the invention

[0005] The purpose of the present invention is to provide a pharmaceutical composition having angiotensin converting enzyme inhibitory effect. Through angiotensin converting enzyme activity influence experiments, it was determined that when the ratio of 3',5-dimethyl hesperidin to 3'-methyl hesperidin was 0.2:1 to 1.6:1, it showed a more effective angiotensin converting enzyme inhibitory effect than using 3'-methyl hesperidin alone.

[0006] The following technical solutions are adopted:

[0007] An application of 3',5-dimethyl hesperidin for preparing a medicine for treating or preventing hypertension.

[0008] Preferably, the invention is used for preparing a drug for inhibiting angiotensin converting enzyme.

[0009] The invention discloses an application of a combination of 3',5-dimethyl hesperidin and 3'-methyl hesperidin for preparing a medicine for treating or preventing hypertension.

[0010] In previous studies, the inventors prepared 3'-methyl hesperidin and 3',5-dimethyl hesperidin with high purity and characterized their structures. Later, they unexpectedly discovered that the two can synergistically produce a strong inhibitory effect on angiotensin-converting enzyme, and have good application prospects in the prevention and treatment of hypertension.

[0011] Preferably, the invention is used for preparing a drug for inhibiting angiotensin converting enzyme.

[0012] The structural formula of 3'5-dimethyl hesperidin is as follows:

[0013]

[0014] The structural formula of 3'-methyl hesperidin is as follows:

[0015]

[0016] Preferably, the molar ratio of the 3',5-dimethyl hesperidin to the 3'-methyl hesperidin is 0.2-1.6:1.

[0017] In the above pharmaceutical composition, when the ratio of 3',5-dimethyl hesperidin to 3'-methyl hesperidin is 0.2:1 to 1.6:1, the ACE inhibitory effect is significantly better than that of 3'-methyl hesperidin alone.

[0018] Preferably, the molar ratio of the 3',5-dimethyl hesperidin to the 3'-methyl hesperidin is 1.2-1.4:1.

[0019] Preferably, the molar ratio of the 3',5-dimethyl hesperidin to the 3'-methyl hesperidin is 1.2:1.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] 1. The present invention first discovered that 3',5-dimethyl hesperidin has angiotensin converting enzyme inhibitory effect;

[0022] 2. 3',5-dimethyl hesperidin and 3'-methyl hesperidin, in a certain ratio, exert a more effective angiotensin-converting enzyme inhibitory effect than using either alone;

[0023] 3. The present invention has discovered for the first time a new use of 3',5-dimethyl hesperidin: it can be used to prepare drugs for lowering blood pressure;

[0024] 4. The present invention has discovered for the first time a new use of the combination of 3',5-dimethyl hesperidin and 3'-methyl hesperidin: it can be used to prepare drugs for lowering blood pressure. DETAILED DESCRIPTION

[0025] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with specific embodiments.

[0026] Example 1

[0027] The angiotensin converting enzyme inhibition test was conducted to compare the differences in the inhibition of enzyme activity between 3'-methyl hesperidin monomer and 3',5-dimethyl hesperidin monomer alone and in combination with mixtures of different proportions. The specific steps include:

[0028] (1) Weigh 19.07 g of borax and 12.37 g of boric acid, respectively, and dissolve them in 1 L of ultrapure water. Measure 325 mL of boric acid solution and 175 mL of borax solution, mix them, adjust the pH to 8.3 with HCl or NaOH, and then add 17.532 g of sodium chloride to make a constant volume of 1 L to prepare a 0.1 mol / L borate buffer solution.

[0029] (2) Weigh a certain amount of hippuryl-histidine-leucine (HHL) and add borate buffer to prepare a solution with a concentration of 5 mmol / L;

[0030] (3) Add 10 mL of borate buffer to the bottle containing 1 U of ACE reagent and shake to completely dissolve it to make a 0.1 U / mL ACE solution.

[0031] (4) Weigh a certain amount of captopril and add borate buffer to prepare a 100 μg / mL solution for use as a positive control.

[0032] (5) Weigh 3'-methyl hesperidin reference substance and 3',5-dimethyl hesperidin reference substance, and prepare the raw material test solution, monomer test solution, and mixed test solution with different ratios (3',5-dimethyl hesperidin: 3'-methyl hesperidin is 0.2, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8);

[0033] (6) Pipette 100 μL of the test solution and 100 μL of 5 mmol / L HHL solution, stir and mix evenly, place in a 37°C water bath, and keep warm for 5 min;

[0034] (7) Add 10 μL of 0.1 U / mL ACE solution, place in a water bath at 37°C to allow it to react for 30 min, and then add 150 μL of 1 mol / L HCl aqueous solution to terminate the reaction;

[0035] (8) Add 1.5 mL of ice-cold ethyl acetate, vortex to mix well, centrifuge at 3500 rpm for 5 min, then take 1 mL of the ester layer and place it in another clean test tube, and dry it at 85°C for 1 h;

[0036] (9) Add 3 mL of ultrapure water and check the absorbance at a wavelength of 228 nm using an ultraviolet spectrophotometer. The test results are shown in Table 1.

[0037] Table 1 Test results of each group

[0038]

[0039]

[0040] Note: A is 3',5-dimethyl hesperidin; B is 3'-methyl hesperidin.

[0041] The test results were statistically processed by SPSS software, and one-way analysis of variance was used. P<0.05 was considered statistically significant. In multiple comparisons (LSD), compared with the positive drug captopril, the inhibitory effect of 3',5-dimethyl hesperidin:3'-methyl hesperidin was significantly different when the ratio was 1.2:1-1.4:1 (P<0.01); compared with 3'-methyl hesperidin alone, the inhibitory effect of 3',5-dimethyl hesperidin:3'-methyl hesperidin was significantly different when the ratio was 0.2:1-1.6:1 (P<0.01). The results calculated according to the combined drug action formula showed that there was an additive effect when the ratio of 3',5-dimethyl hesperidin:3'-methyl hesperidin was 1.2:1.

[0042] The above disclosure is only the preferred embodiment of the present invention, which certainly cannot be used to limit the scope of the present invention. Therefore, equivalent changes made according to the claims of the present invention are still within the scope of the present invention.

Claims

1. An application of 3',5-dimethyl hesperidin, characterized in that: Used in the preparation of medicines for treating or preventing hypertension.

2. The use of 3',5-dimethyl hesperidin according to claim 1, characterized in that: Used to prepare drugs that inhibit angiotensin converting enzyme.

3. The use of 3',5-dimethyl hesperidin according to claim 1, characterized in that: The 3',5-dimethyl hesperidin includes the following structural formula:

4. A pharmaceutical composition containing 3',5-dimethyl hesperidin, characterized in that: Also included is 3'-methylhesperidin.

5. The pharmaceutical composition according to claim 4, characterized in that The 3'-methyl hesperidin comprises the following structural formula:

6. The pharmaceutical composition according to claim 4, characterized in that: Used in the preparation of medicines for treating or preventing hypertension.

7. The pharmaceutical composition according to claim 4, characterized in that: Used to prepare drugs that inhibit angiotensin converting enzyme.

8. The pharmaceutical composition according to claim 4, characterized in that: The molar ratio of the 3',5-dimethyl hesperidin to the 3'-methyl hesperidin is 0.2-1.6:

1.

9. The pharmaceutical composition according to claim 4, characterized in that: The molar ratio of the 3',5-dimethyl hesperidin to the 3'-methyl hesperidin is 1.2-1.4:

1.

10. The pharmaceutical composition according to claim 4, characterized in that: The molar ratio of the 3',5-dimethyl hesperidin to the 3'-methyl hesperidin is 1.2:1.

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