TWI680762B - Peptide for inhibiting angiotensin converting enzyme, hydrolysate of cassia seeds, food composition or medical composition including the peptide and a use of the peptide thereof - Google Patents

Abstract

A peptide for inhibiting angiotensin converting enzyme (ACE) can be used as a new solution for patients who have side effects due to conventional ACE inhibitors. The peptide has an amino acid sequence set forth as SEQ ID NO: 1. A hydrolyze of Cassia seeds, a food composition or a medical composition including the peptide, and a use of the peptide in the manufacture of an ACE inhibitor are also disclosed.

Description

A peptide for inhibiting vasopressin converting enzyme, and a decision containing the peptide Mingzi hydrolysate, food composition or pharmaceutical composition, and use of the peptide

The present invention relates to a peptide, especially a peptide for inhibiting vasopressin converting enzyme; the present invention also relates to a cassia hydrolysate containing the peptide, and the peptide for preparing vasopressin converting enzyme inhibition Use of the agent.

Angiotensin I converting enzyme (ACE) can convert angiotensin I into angiotensin II inhibitor with strong vasoconstriction effect, thus inhibiting angiogenesis Metotonin-converting enzyme can reduce the production of vasopressin II, achieve vasodilation, blood volume reduction and blood pressure drop, etc. It is often used as a medicine for hypertension and can also be used to treat acute myocardial (acute myocardial) infarction), cardiac failure, diabetes nephropathyl, etc.

Conventional ACE inhibitors include captopril, enalapril, fosinopril, lisinopril, etc. Conventional use of vasopressin-converting enzyme inhibitors may cause side effects such as dry cough, and patients with poor renal function are more likely to cause symptoms of hyperkalemia. It is still necessary to provide a peptide for inhibiting vasopressin converting enzyme to improve the above problems.

In order to solve the above problems, an object of the present invention is to provide a peptide for inhibiting vasopressin converting enzyme, which has good vasopressin converting enzyme inhibitory activity.

The next object of the present invention is to provide a cassia seed hydrolysate, which contains the peptide as described above.

Still another object of the present invention is to provide a use of a peptide for preparing an inhibitor of vasopressin converting enzyme.

The peptide for inhibiting vasopressin converting enzyme of the present invention has the amino acid sequence described in SEQ ID NO:1. The present invention has as SEQ ID NO: amino acid sequence of the peptide of FIG 1, having a good vascular vasopressin converting enzyme inhibitory activity (half-vascular vasopressin converting enzyme inhibitory concentration (IC ₅₀₎ of about 16.83±0.90μM), so it can be used as an active ingredient to inhibit vasopressin-converting enzyme, and can be used to prepare vasopressin-converting enzyme inhibitors. For patients who induce side effects, a new option for vasopressin converting enzyme inhibitors is provided for the efficacy of the present invention.

The cassia seed hydrolysate, food composition, or pharmaceutical composition of the present invention contains the peptide as described above. By using the peptide as described above, the Cassia seed hydrolysate, the food composition or the pharmaceutical composition can be used as an vasopressin converting enzyme inhibitor, which is the effect of the present invention.

The use of the peptide of the present invention as described above is to prepare an vasopressin converting enzyme inhibitor, wherein the peptide can be administered to a desired individual to inhibit the activity of vasopressin converting enzyme, In order to improve the symptoms of hypertension, atherosclerosis, thrombosis, heart failure, diabetes, diabetic kidney complications or cardiac insufficiency of the desired individual; preferably, the peptide can be administered to the institute orally Need individual. In this way, the peptide can be used in the desired individual Can inhibit the activity of vasopressin converting enzyme in the desired individual, and can improve the hypertension, atherosclerosis, thrombosis, heart failure, diabetes, diabetic kidney complications or cardiac insufficiency of the desired individual Symptoms are the efficacy of the present invention.

[Figure 1] This is the result of analyzing the amino acid sequence of the peptide of the present invention by high-resolution mass spectrometry.

[Figure 2] It is a scatter plot of the percent inhibition of vasopressin converting enzyme of the peptide of the present invention at different concentrations.

[Figure 3] This is a line chart of blood pressure changes at different time points in hypertensive rats fed with the peptide of the present invention.

In order to make the above-mentioned and other objects, features and advantages of the present invention more obvious and understandable, the preferred embodiments of the present invention are described below in conjunction with the accompanying drawings, which are described in detail as follows: A peptide that inhibits angiotensin converting enzyme has an amino acid sequence as shown in SEQ ID NO: 1, the peptide can be artificially synthesized, or extracted and purified from a cassia seed sample (eg, Cassia obtusifolia L.) seeds or Cassia tora L. seeds), which is understood by those with ordinary knowledge in the technical field to which the invention belongs, and is not limited herein.

In this embodiment, the peptide is extracted and purified from the cassia seed sample as follows: the seeds of the cassia are ground into powder, and then the protein contained therein is precipitated with trichloroacetic acid, followed by thermolysin ) Hydrolysis treatment (the reaction is carried out at 60°C, pH 8.3 for 16 hours, and heated at 95°C for 5 minutes to terminate the hydrolysis reaction), and finally the thermophilic protease is removed with a 3kDa ultrafiltration membrane, leaving only a molecular weight of less than 3kDa The cassia hydrolysate is subjected to subsequent purification Chemical conversion (for example, by reverse phase high performance liquid chromatography or strong cation exchange chromatography).

The peptide has good vasopressin-converting enzyme inhibitory activity, and its half-inhibitory concentration (IC ₅₀ ) is about 16.83±0.90 μM, so it can be used as an active ingredient for inhibiting vasopressin-converting enzyme. The peptide is used to prepare an vasopressin converting enzyme inhibitor. The peptide is combined with a pharmaceutically acceptable carrier or excipient to form a pharmaceutical composition or a food composition, and can be prepared into any convenient edible The type, such as tablets, capsules, powders, granules or liquids, etc., can be taken orally by a desired individual in a form suitable for consumption.

In addition, the peptide can be administered to the desired individual. For example, the peptide can be administered to the desired individual at a dose of more than 1 mg (>1 mg/kg (BW)) per kg of body weight per day. , So that the peptide can act in the body of the desired individual, and can achieve the effects of vasodilation, blood volume reduction and blood pressure reduction.

In order to confirm that the peptide used to inhibit angiotensin-converting enzyme does indeed inhibit the activity of angiotensin-converting enzyme and can effectively reduce the blood pressure value of the desired individual, the following test was carried out:

(A) The amino acid sequence of the peptide of the present invention

In this experiment, the amino acid sequence was analyzed by high-resolution mass spectrometry (Q Exactive ^TM Hybrid Quadrupole-Orbitrap), and the analysis results were converted to software (Mascot Distiller), and then de novo sequencing. Sequenced, the result is shown in Figure 1, and the peptide of the present invention does have the amino acid sequence shown in SEQ ID NO:1.

(B) Inhibitory activity of the peptide of the present invention on vasopressin converting enzyme

In this test, different concentrations of peptides were co-reacted with vasopressin-converting enzyme and vasopressin-converting enzyme substrate (Hippury-L-Histidyl-L-Leucine, HHL), and then the hippuric acid was detected ( The production amount of hippury acid (HA) was used to evaluate the inhibitory effect of the peptide of the present invention on vasopressin converting enzyme. The results are shown in Figure 2 and the semi-inhibitory concentration of the peptide of the present invention (IC ₅₀ ) Is about 16.83±0.90 μM, showing that the peptide of the present invention has good vasopressin converting enzyme inhibitory activity.

(C) Blood pressure lowering effect of the peptide of the present invention

In this experiment, spontaneously hypertensive rats weighing approximately 250 grams were selected, and rats in groups C1 and C2 were fed with angiotensin-converting enzyme inhibitors as shown in [Table 1], and then dosed at different time points Measure tail blood pressure. In this test, an equal volume of water was used as Group C0.

As shown in Figure 3, the blood pressure of rats in group C0 fed with water did not change significantly within 24 hours, and the blood pressure of rats in group C1 fed with the peptide of the present invention reached the lowest level at 6 hours At this point, the change in systolic blood pressure is about 17 millimeters of mercury (mmHg), showing that the peptide of the present invention can indeed reduce the blood pressure of hypertensive rats.

In summary, the peptide of the present invention having the amino acid sequence shown in SEQ ID NO: 1 has good angiotensin-converting enzyme inhibitory activity (semi-inhibitory concentration of angiotensin-converting enzyme (IC ₅₀ ) Is about 16.83±0.90μM), so it can be used as an active ingredient for inhibiting vasopressin converting enzyme, and can be used in the preparation of vasopressin converting enzyme inhibitors, for the administration of conventional vasopressin converting enzyme Patients with inhibitors that induce side effects provide a new option for vasopressin-converting enzyme inhibitors for the efficacy of the present invention.

Furthermore, since the peptide of the present invention can be applied to prepare an vasopressin converting enzyme inhibitor, it can be administered to a desired individual so that the peptide can function in the desired individual. Inhibiting the activity of the vasopressin converting enzyme in the desired individual can improve the symptoms of hypertension, atherosclerosis, thrombosis, heart failure, diabetes, diabetic kidney complications or cardiac insufficiency in the desired individual. The effect of the invention.

Although the present invention has been disclosed using the above-mentioned preferred embodiments, it is not intended to limit the present invention. Anyone who is familiar with this art without departing from the spirit and scope of the present invention still makes various changes and modifications to the above-mentioned embodiments. The technical scope of the invention is protected, so the scope of protection of the present invention shall be subject to the scope defined in the appended patent application.

<110> Daren University of Science and Technology

<120> Peptide for inhibiting vasopressin converting enzyme, cassia hydrolysate containing the peptide, food composition or pharmaceutical composition, and use of the peptide

<140> TW 107121351

<141> 2018-06-21

<160> 1

<170> PatentIn version 3.5

<210> 1

<211> 6

<212> PRT

<213> Cassia obtusifolia L. (Cassia)

<400> 1

Claims (6)

A peptide for inhibiting vasopressin converting enzyme has an amino acid sequence as described in SEQ ID NO:1. A cassia seed hydrolysate comprising the peptide described in item 1 of the patent application scope; wherein, the cassia seed hydrolysate is prepared by a method comprising the following steps: providing a cassia seed powder; precipitating the cassia with trichloroacetic acid The protein in the seed powder was subjected to a hydrolysis reaction with thermophilic protease at 60°C and pH 8.3 for 16 hours, and heated at 95°C for 5 minutes to terminate the hydrolysis reaction; and the 3kDa ultrafiltration membrane was used to remove the thermophile Protease to obtain cassia hydrolysate with molecular weight less than 3kDa. A food composition comprising the peptide described in item 1 of the patent application. A pharmaceutical composition comprising the peptide described in item 1 of the patent application. The use of a peptide as described in item 1 of the patent application is to prepare an vasopressin converting enzyme inhibitor, wherein the peptide is administered to a desired individual to inhibit vasopressin converting enzyme Activity, and thereby improve the symptoms of hypertension, atherosclerosis, thrombosis, heart failure, diabetes, diabetic kidney complications or cardiac insufficiency in the desired individual. The use of the peptide as described in item 5 of the patent application scope, wherein the peptide is administered to the desired individual orally.

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