CN113616776A

CN113616776A - Application of blocker C23 in preparation of medicine for treating myocardial infarction

Info

Publication number: CN113616776A
Application number: CN202110958395.6A
Authority: CN
Inventors: 李勇男
Original assignee: Lanzhou University Second Hospital
Current assignee: Lanzhou University Second Hospital
Priority date: 2021-08-20
Filing date: 2021-08-20
Publication date: 2021-11-09

Abstract

The invention discloses an application of a blocker C23 in preparing a medicine for treating myocardial infarction, wherein C23 is firstly used for preparing the medicine for treating myocardial infarction, and the effect of C23 in controlling heart disease is verified on an animal model. Particularly, C23 can reduce the content of CK-MB, cTnT and cTnI in the serum of a rat with coronary artery ligation and the myocardial infarction area, remarkably improve the 15-day cardiac function and reduce NT-pro BNP and myocardial fibrosis. In addition, the C23 of the invention has the advantages of low toxicity, small molecular weight, low immunogenicity, high drugability and the like when used as a pharmaceutical ingredient.

Description

Application of blocker C23 in preparation of medicine for treating myocardial infarction

Technical Field

The invention belongs to the technical field of protein engineering, and particularly relates to application of a blocker C23 in preparation of a medicine for treating myocardial infarction.

Background

Myocardial infarction is the partial necrosis of a portion of the myocardium due to severe, persistent ischemic hypoxia resulting from partial or complete occlusion of the coronary arteries. Epidemiological studies show that at present, more than 2.9 hundred million patients with cardiovascular diseases in China have more than 1100 million patients with coronary heart diseases, and 50% of cardiovascular disease deaths are caused by myocardial infarction. After myocardial infarction, innate immunity is rapidly activated to generate strong and transient inflammatory response, dead cells and extracellular matrix fragments in the infarct area are cleared, and then the proliferation stage is carried out. During the proliferative phase, monocyte and macrophage subpopulations secrete growth factors, recruiting and activating mesenchymal repair cells (mainly myofibroblasts and vascular cells). Myofibroblasts secrete large amounts of extracellular matrix proteins, thereby retaining the left ventricular intact structure. Most repair apoptosis means the end of the proliferative phase, collagen fiber scarring.

Tissue damage produces endogenous signal molecules that activate the innate immune system, and these signal molecules belong to the family of risk-associated pattern molecules (DAMPS). CIRBP is one of them, which mediates post-myocardial ischemic inflammatory injury through Toll-like receptors (TLRs).

Innate immunity recognizes danger signals through TLRs, and of the TLR family, TLR2 and TLR4 play important mediating roles in post-myocardial inflammatory responses.

Research shows that the interaction between CIRBP and TLR4 plays a key role in post-myocardial inflammatory reaction to aggravate ischemic injury. It is desirable to have the ability to specifically block the binding of CIRBP to TLR4 and reduce post-myocardial inflammatory injury. Currently, intervention strategies and therapeutic drugs that can effectively reduce myocardial infarction injury are very limited. Therefore, the invention can provide a candidate drug with clinical application value for the treatment of myocardial infarction injury.

Disclosure of Invention

In view of the above, the present invention provides an application of blocker C23 in the preparation of a medicament for treating myocardial infarction, which aims at the problems existing in the prior art.

In order to solve the technical problems, the technical scheme adopted by the invention is as follows:

in a first aspect of the invention, the invention provides an application of a blocker C23 in preparing a medicament for treating myocardial infarction.

The invention further consists in:

the blocking agent C23 can reduce the content of creatine kinase CK-MB and troponin cTnT and cTnI in the blood serum of a rat with a ligated coronary artery, obviously improve the cardiac function for 15 days and reduce NT-pro BNP and myocardial fibrosis.

The invention further consists in:

the blocking agent C23 can reduce myocardial infarction area of rat, and has protective effect on myocardial ischemia.

In a second aspect of the invention, a medicament for treating myocardial infarction, comprises the blocker C23.

The invention further consists in:

the amino acid sequence of the blocker C23 is as follows: GRGFSRGGGDRGYGG are provided.

The invention further consists in:

the administration route of the medicament comprises intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration or transdermal administration.

The invention further consists in:

the dosage form of the medicine is tablets, granules, pills, powder, capsules, injections, oral liquid, ophthalmic preparations or external preparations.

The invention can obtain the following technical effects:

the invention discloses an application of a specific blocker C23 of CIRBP and TLR4 in preparing a medicament for treating myocardial infarction, which is proved by experiments and comprises the following steps: the blocking agent C23 can effectively reduce the injury after myocardial infarction.

The invention firstly uses C23 in preparing the medicine for treating myocardial infarction, and the effect of C23 in controlling heart disease is verified on an animal model. Particularly, C23 can improve the activity of myocardial cells under an anoxic state, obviously inhibit the activation of apoptosis-related protein and the accumulation of active oxygen, reduce the content of CK-MB, cTnT and cTnI in the blood serum of a rat with coronary artery ligation, obviously improve the cardiac function for 15 days and reduce NT-proBNP and myocardial fibrosis. In addition, the C23 of the invention has the advantages of low toxicity, small molecular weight, low immunogenicity, high drugability and the like when used as a pharmaceutical ingredient.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention and not to limit the invention. In the drawings:

FIG. 1 shows the test results of the effect of C23 polypeptide on the content of troponin cTnT and cTnI in the serum of SD rat with myocardial infarction;

FIG. 2 shows the test results of the effect of C23 polypeptide on the content of CK-MB in serum of myocardial infarction SD rats;

FIG. 3 shows the test results of the effect of C23 polypeptide on the myocardial infarction area of groups of myocardial infarction SD rats;

FIG. 4 shows the results of a test of the effect of C23 polypeptide on the cardiac color ejection fraction (EF%) of 15 days of myocardial infarction SD rats;

FIG. 5 shows the results of a test of the effect of C23 polypeptide on day 15 serum NT-proBNP of groups of myocardial infarction SD rats;

fig. 6 is a test result of the effect of C23 polypeptide on myocardial tissue fibrosis of groups of myocardial infarction SD rats for 15 days.

Detailed Description

The following embodiments are described in detail with reference to the accompanying drawings, so that the implementation process of the present invention for solving the technical problems and achieving the technical effects by applying technical means can be fully understood and implemented.

The embodiment of the invention relates to application of polypeptide for treating myocardial infarction, in particular to application of C23 in a medicament for treating myocardial infarction, wherein the amino acid sequence of C23 is GRGFSRGGGDRGYGG.

The "polypeptide" referred to in the examples of the present invention is a compound in which a-amino acids are linked together by peptide bonds, and is an intermediate product of proteolysis. The compounds obtained by dehydration condensation of two amino acid molecules are called dipeptides, and similarly, tripeptides, tetrapeptides, pentapeptides, and the like are also included. The compound formed by dehydration condensation of 10-100 amino acid molecules is usually called polypeptide.

It is understood that the administration route of the drug in the embodiment of the present invention is not particularly limited, and includes, but is not limited to, intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration, transdermal administration, etc.

It is to be understood that the dosage form of the medicament is not particularly limited in the embodiments of the present invention, and includes, but is not limited to, tablets, granules, pills, powders, capsules, injections, oral liquids, ophthalmic preparations, external preparations, and the like.

The invention firstly uses C23 as a medicament for treating myocardial infarction, and the effect of C23 on controlling heart disease is verified on an animal model. Particularly, C23 can improve the activity of myocardial cells under an anoxic state, obviously inhibit the activation of apoptosis-related protein and the accumulation of active oxygen, and reduce the content of CKMB, cTnT and cTnI in the blood serum of a rat with coronary artery ligation and the myocardial infarction area. In addition, the C23 of the invention has the advantages of low toxicity, small molecular weight, low immunogenicity, high drugability and the like when used as a pharmaceutical ingredient.

The first embodiment is as follows: the blocking agent C23 can reduce the content of creatine kinase CK-MB, troponin cTnT and cTnI in the blood serum of a rat with a ligated coronary artery, and the myocardial infarction area of the rat.

(1) Experimental animals: male SD rats, SPF grade, body weight: 250g-350g, provided by Wintonlihua Limited liability company, and raised in laboratory animal center of Lanzhou university.

(2) Grouping experiments: SD rats were randomly divided into four groups: the preparation method comprises 6 pseudo-surgery groups, 6 modeling groups, 6 pseudo-surgery and C23 polypeptide groups, 6 modeling C23 polypeptide groups, 8mg/Kg administration of C23 polypeptide, preparation of injection by using normal saline as a solvent, and administration by tail vein injection.

(3) Coronary artery ligation of SD rat

SD rats were anesthetized with 3% sevoflurane, fixed in the supine position, the chest wall was cut along the fifth left intercostal and the ribs of the fourth and fifth costal bones were cut 2mm from the left edge of the sternum, and the pericardium was cut to expose the heart.

The left anterior descending artery is threaded through a No. 5-0 suture line to ligate the blood vessel of the left anterior descending artery, the heart is put back into the thoracic cavity, the air in the chest is exhausted by hand as much as possible, and the thoracic cavity is closed quickly. The whole process is completed within 30 seconds.

Myocardial ischemia is caused after the coronary artery of the rat is ligated, and the coronary artery of the sham operation group is not ligated. Administration was performed by tail vein injection 30 minutes after coronary artery ligation. 3 hours later, 3ml of blood is taken from femoral artery, supernatant is taken by conventional centrifugation, and the levels of serum creatine kinase CK-MB, troponin cTnT and cTnI are measured; carrying out TTC staining on heart tissues, and observing the size of a myocardial infarction area; after 15 days, performing heart color ultrasonography, checking the cardiac ejection fraction, taking 3ml of blood, conventionally centrifuging, taking supernatant, measuring the level of serum NT-proBNP, taking heart tissue, performing Masson staining, and observing the degree of myocardial fibrosis.

(4) Results of the experiment

FIG. 1 shows the test results of the effect of C23 polypeptide on the content of troponin (cTnT, cTnI) in the serum of myocardial infarction SD rats, and FIG. 2 shows the test results of the effect of C23 polypeptide on the content of creatine kinase CK-MB in the serum of myocardial infarction SD rats. As can be seen from FIG. 1 and FIG. 2, the serum creatine kinase CK-MB and troponin cTnT, cTnI content of the model-made SD rat is significantly higher than that of the sham-operated group (P <0.01), which indicates that myocardial ischemia of the rat is caused after coronary artery ligation to generate myocardial damage, so that creatine kinase CK-MB and troponin cTnT, cTnI are released into the blood; the contents of creatine kinase CK-MB, troponin cTnT and cTnI in the rat serum of the model group added with C23 polypeptide are obviously reduced compared with the model group (P is less than 0.01).

In conclusion, the C23 polypeptide can reduce the content of creatine kinase CK-MB, troponin cTnT and cTnI in the serum of rats with myocardial ischemia-reperfusion injury.

Fig. 3 is a test result of the effect of C23 polypeptide on myocardial infarction area of various groups of myocardial infarction SD rats. As can be seen from FIG. 3, the myocardial infarction area of the rats in the model group is significantly higher than that of the sham operation group (P <0.01), while the myocardial infarction area of the rats in the model group added with C23 polypeptide is significantly reduced compared with that in the model group (P < 0.01).

Fig. 4 is a test result of the effect of C23 polypeptide on 15-day cardiac color hypersecretion fraction (EF%) of various groups of myocardial infarction SD rats. As shown in FIG. 4, the 15-day cardiac color hypersecretion fraction (EF%) of the rats in the model group was significantly lower than that in the sham-operated group (P <0.01), while the 15-day cardiac color hypersecretion fraction (EF%) of the rats in the model group added with C23 polypeptide was significantly higher than that in the model group (P < 0.01).

FIG. 5 shows the results of a test of the effect of C23 polypeptide on day 15 serum NT-proBNP of groups of myocardial infarction SD rats. As shown in FIG. 5, the serum NT-proBNP of the model-made rat is significantly higher than that of the sham-operated rat (P <0.01) in 15 days, while the serum NT-proBNP of the model-made rat added with C23 polypeptide is significantly lower than that of the model-made rat (P < 0.01).

Fig. 6 is a test result of the effect of C23 polypeptide on myocardial tissue fibrosis of groups of myocardial infarction SD rats for 15 days. As shown in FIG. 6, the fibrosis of myocardial tissue of rats modeled after 15 days is significantly higher than that of the sham-operated group (P <0.01), while the fibrosis of myocardial tissue of rats modeled after adding C23 polypeptide is significantly reduced compared with that of the modeled group (P < 0.01).

In conclusion, the C23 polypeptide can reduce the myocardial infarction area of rats and has obvious protective effect on myocardial ischemia.

While the foregoing description shows and describes several preferred embodiments of the invention, it is to be understood, as noted above, that the invention is not limited to the forms disclosed herein, but is not to be construed as excluding other embodiments and is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as expressed herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. An application of blocker C23 in preparing medicine for treating myocardial infarction is disclosed.

2. The use according to claim 1, wherein the blocking agent C23 is capable of reducing the content of creatine kinase CK-MB and troponin cTnI, cTnT in the serum of coronary artery ligated rats, significantly improving the 15-day cardiac function and reducing NT-proBNP and myocardial fibrosis.

3. The use of claim 1, wherein said blocking agent C23 is capable of reducing the infarct size in rat myocardium and has protective effect on myocardial ischemia.

4. A medicine for treating myocardial infarction is characterized by comprising a blocking agent C23.

5. The medicament of claim 4, wherein the amino acid sequence of the blocker C23 is: GRGFSRGGGDRGYGG are provided.

6. The medicament of claim 5, wherein the route of administration of the medicament comprises intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration or transdermal administration.

7. The medicament of claim 5, wherein the medicament is in the form of tablets, granules, pills, powder, capsules, injections, oral liquid, ophthalmic preparations or external preparations.