CN111110672A - Application of DYZ-9 in preparation of anti-myocardial hypertrophy products - Google Patents

Abstract

The invention discloses an application of DYZ-9 in preparation of a myocardial hypertrophy resistant product, and compared with the prior art, the invention has the following advantages: (1) the DYZ-9 researched by the invention can be synthesized by a known method, has a mature preparation process, can provide a reliable compound source, and has the advantages of material source reliability, low production cost, novel action mechanism, good market prospect and the like. (2) The invention can inhibit the phosphorylation of AKT and adenosine cyclophosphate effect element binding protein caused by Isoproterenol (ISO), reduce the levels of markers ANP and BNP of myocardial hypertrophy and cardiac weight index, and improve myocardial hypertrophy. (3) The invention is beneficial to the research and development of novel anti-myocardial hypertrophy medicines, can bring direct economic benefits, can reduce the possibility that the myocardial hypertrophy is further worsened to form heart failure and malignant arrhythmia, and can save a large amount of sanitary expenses in the long term.

Description

Application of DYZ-9 in preparation of anti-myocardial hypertrophy products

Technical Field

The invention belongs to the technical field of medicines, and relates to application of DYZ-9 in preparation of a product for resisting myocardial hypertrophy.

Background

The seed of the mahogany rosewood as a traditional medicine is subjected to treatment verification of local residents in the long years in south-east Asia countries, and the seed is proved to possibly have certain anti-inflammatory and other effects. However, after it is introduced into southern provinces of China, such as Hainan province, it is only used as a street tree for viewing and admiring at present, and the value of the fruit is not fully utilized. If the device can be reasonably developed and utilized, great value is created. In Indonesia, the seed of mahogany is used as a traditional herbal medicine for treating diseases such as hypertension and diabetes, but the exact effective ingredients in the seed are lack of systematic study.

DYZ-9 (swiitenine) with molecular formula C₃₂H₄₀O₉The structural formula is shown as formula I (temporarily marked as DYZ-9 to distinguish from similar products with the same name on the market, and the CAS No:4802-04-4 thereof) is separated from peach blossom seeds for the first time in 1990.

Few studies of successful isolation of DYZ-9 have demonstrated that several substances extracted have therapeutic effects on the corresponding diseases, and experimental studies in which DYZ-9 has been reported have shown that it can treat diabetes and platelet aggregation, and the effects on other diseases are not well understood.

The β adrenoceptor blockers such as betalaks, atenolol and the like have been widely used in clinical treatment of diseases such as heart failure and hypertension, but the effect of reversing ventricular hypertrophy is not good, and studies on DYZ-9 in cardiac hypertrophy are not reported.

Disclosure of Invention

The purpose of the invention is as follows: the technical problem to be solved by the invention is to provide a potentially better therapeutic drug for treating myocardial hypertrophy (especially for diabetes patients). Application of DYZ-9 in preparing products for resisting myocardial hypertrophy.

In order to solve the technical problems, the invention discloses application of DYZ-9 in preparation of a myocardial hypertrophy resistant product.

Wherein, the chemical structure of DYZ-9 is shown as formula I,

preparation method of the DYZ-9

(1) Sun drying seed of heartwood of Prunus davidiana Maxim, pulverizing (1.6kg), extracting with 95% ethanol at room temperature for 3 times (each for 7 days), and recovering ethanol under reduced pressure until no ethanol smell is present to obtain ethanol extract;

(2) dispersing the ethanol extract in water to obtain suspension, sequentially extracting with ethyl acetate and n-butanol for 3 times, respectively, and concentrating under reduced pressure to obtain ethyl acetate extract (110.0g) and n-butanol extract (19.0 g).

(3) Selecting ethyl acetate extract (110.0g), carrying out MCI column chromatography, carrying out gradient elution by 50-100% methanol water solution, finally carrying out acetone elution, carrying out thin layer chromatography detection, and combining the same fractions to obtain 4 components (Fr.1-Fr.4).

(5) And performing reduced pressure column chromatography on the component Fr.2, and repeatedly eluting by reverse phase column chromatography (methanol-water), Sephadex LH-20 gel column chromatography (methanol, chloroform-methanol) and normal phase silica gel column chromatography to obtain DYZ-9. Reference: plant activation Inhibitors from the feeds of Swietariamagic, Inhibition of Inhibition and in vivo of plasmid-activation factors Induced to Swietentine and Swietentine, Hisao Eimo. Yukio.Yukio. Iri, et al.plant mecl.57(1991):56-58.

Wherein the myocardial hypertrophy is isoproterenol induced myocardial hypertrophy.

Wherein, the anti-myocardial hypertrophy is effective prevention, alleviation and treatment of myocardial hypertrophy.

Wherein, the anti-myocardial hypertrophy is to reduce the level of Atrial Natriuretic Peptide (ANP) and Brain Natriuretic Peptide (BNP) which are markers of myocardial hypertrophy and the cardiac weight index and improve the myocardial hypertrophy by inhibiting the phosphorylation of protein kinase B (AKT) and adenosine cyclophosphate effect element binding protein (CREB) caused by Isoproterenol (ISO) by DYZ-9.

Among them, ISO can activate its downstream PI3K/AKT and PKA/CREB signaling pathways through G protein-coupled receptors to cause cardiac hypertrophy.

Wherein the product is any one of medicine, food and health care product. The dosage form of the product is preferably any one of powder, granules, tablets and capsules.

An anti-myocardial hypertrophy drug monomer comprises effective amount of DYZ-9.

An anti-myocardial hypertrophy pharmaceutical composition comprises an effective amount of DYZ-9 or pharmaceutically acceptable salts and hydrates thereof and pharmaceutically acceptable carriers, auxiliary agents or compositions thereof.

The dosage form of the drug monomer and the composition is preferably any one of powder, granules, tablets and capsules.

Has the advantages that: compared with the prior art, the invention has the following advantages:

(1) the DYZ-9 researched by the invention can be synthesized by a known method, has a mature preparation process, can provide a reliable compound source, and has the advantages of material source reliability, low production cost, novel action mechanism, good market prospect and the like.

(2) The invention can inhibit the phosphorylation of AKT and adenosine cyclophosphate effect element binding protein caused by Isoproterenol (ISO), reduce the levels of markers ANP and BNP of myocardial hypertrophy and cardiac weight index, improve myocardial hypertrophy and reverse ventricular hypertrophy.

(3) The invention is beneficial to the research and development of novel anti-myocardial hypertrophy drugs, can bring direct economic benefits, and brings advantages for the treatment of patients suffering from various related diseases by reported treatment functions of diabetes and other diseases. Secondly, the medicine can reduce the possibility that the cardiac hypertrophy is further worsened to form heart failure and malignant arrhythmia, and can save a great deal of sanitary expenses in the long term.

(4) The invention improves the clinical cure rate and the long-term survival rate of patients with cardiac hypertrophy, relieves the pain and the mental stress of patients and improves the labor productivity of patients.

Drawings

Fig. 1 shows the comparison of the super-junction fruits between groups.

In FIG. 2, A is HE staining contrast among groups; b is the comparison of the cross-sectional area of the myocardial cells among the groups.

FIG. 3 is a comparison of the expression levels of ANP and BNP mRNA in cardiac tissues among groups.

In FIG. 4, A is H9C2 α -SMA immunofluorescence among groups, and B is a comparison of the mean area of myocardium among groups.

FIG. 5 is a graph showing the comparison of the expression levels of mRNA for ANP and BNP in each group of H9C 2.

In FIG. 6, A is the western blot level in each set of H9C 2; b is the comparison of the p-AKT/AKT and p-CREB/CREB protein expression levels of each group.

In FIG. 7, A is the western blot level in heart tissue of each group of mice; b is the comparison of the p-AKT/AKT protein expression levels of each group.

Detailed Description

The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.

Example 1: establishment of myocardial hypertrophy animal model and detection of DYZ-9 drug effect on myocardial hypertrophy resistance

Materials and methods

1. Collecting and preparing a specimen: (1) male ICR mice of 8 weeks of age were selected and the drug isoproterenol (ISO, 5mg/kg/day) was continuously pumped subcutaneously, while DYZ-9(1, 10mg/kg/day) was given a single daily intraperitoneal injection for 2 weeks. (2) All mice were divided into 4 groups, i.e., a Control group (Control group, healthy mice without cardiac hypertrophy), isoproterenol group (ISO group), low dose DYZ-9 treatment group (DYZ-91 mg/kg + ISO group), and high dose DYZ-9 treatment group (DYZ-910 mg/kg + ISO group).

2. Measuring the thickness of the left ventricle and the weight index of the left ventricle by using the echocardiogram of the heart; heart and body weight mass were measured after sacrifice of the animals.

3. The morphology of the heart histology was observed with hematoxylin-eosin (H & E) staining.

4. And detecting the mRNA expression levels of the heart ANP and the BNP by a real-time quantitative PCR method.

Second, experimental results

DYZ-9 reverses ISO-induced myocardial hypertrophy. After 2 weeks of ISO treatment, there was a significant increase in HW, HW/BW, IVSd, LVPWd, LV Mass, all reversed by the high dose DYZ-9 treatment group (see figure 1, table 1).

Table 1: comparison of cardiac weight and wall thickness of the cardiac chamber between groups

Note: HW-heart weight, BW-body weight, IVSd-ventricular septal thickness, LVPWd-left ventricular free wall thickness, lvmas-left ventricular weight index.

#: in contrast to Control group, P<0.05; compares to ISO, P<0.05. For measuring data

Is represented by P<0.05 the difference was considered statistically significant.

2. The heart H & E staining also suggested that the high dose DYZ-9 treatment group significantly improved the size of the heart and the cross-sectional area of the myocardial fibers as compared to the ISO group (see fig. 2A, B).

DYZ-9 reverses the increase in mRNA expression of ANP, BNP in myocardial tissues of mice after ISO treatment (see FIG. 3).

Example 2: establishment of isolated myocardial cell hypertrophy model and research on DYZ-9 anti-myocardial hypertrophy action mechanism

Materials and methods

1. An ISO (10 mu M) in vitro stimulation rat myocardial cell line (H9C2) is selected to establish an in vitro myocardial cell hypertrophy model. Treatment group DYZ-9 (20. mu.M, 50. mu.M, 100. mu.M) was incubated with ISO for 24 h.

2, α -SMA immunofluorescence to observe H9C2 cell morphology.

3. And detecting the mRNA expression levels of ANP and BNP of the H9C2 cells by a real-time quantitative PCR method.

And 4, detecting the expression levels of p-Akt, Akt, p-CREB and CREB proteins by using Western blot.

Second, experimental results

1. Immunofluorescence α -SMA assay suggested that ISO stimulation could significantly increase H9C2 cell volume, while DYZ-9 could reverse it (FIGS. 4A, B).

2. Compared with a normal control group, ISO (10 mu M) can obviously increase the mRNA expression level of ANP and BNP in H9C 2. The DYZ-9 group significantly improved the increase in expression levels of ANP and BNP caused by ISO (FIG. 5).

Phosphorylation levels of Akt, CREB were significantly elevated following ISO stimulation, while treatment with DYZ-9 reversed the phosphorylation levels of Akt and CREB (fig. 6A, B).

Example 3: validation of DYZ-9 action targets at Whole animal level

Materials and methods

Male ICR mice of 8 weeks old are selected, continuously pumped with the drug isoproterenol subcutaneously and given with DYZ-9(1, 10mg/kg/day) once a day for intraperitoneal injection, and after 2 weeks of death, Western blot is used for detecting the p-Akt and Akt protein expression level of the mice.

Second, experimental results

DYZ-9 treatment significantly reversed ISO resulting in elevated phosphorylation levels of Akt and CREB in mouse heart tissue (FIGS. 7A, B).

The present invention provides the idea and method of the application of DYZ-9 in the preparation of anti-myocardial hypertrophy products, and the method and the way to implement the technical solution are many, and the above description is only the preferred embodiment of the present invention, it should be noted that for those skilled in the art, several modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention. All the components not specified in the present embodiment can be realized by the prior art.

Claims (9)

1. The application of DYZ-9 in preparing products for resisting myocardial hypertrophy; wherein, the chemical structure of DYZ-9 is shown as formula I,

   
2. 2. the use of claim 1, wherein the cardiac hypertrophy is isoproterenol induced cardiac hypertrophy.
3. 3. The use of claim 1, wherein the anti-myocardial hypertrophy is improvement of myocardial hypertrophy by decreasing the levels of atrial natriuretic peptide and brain natriuretic peptide, markers of myocardial hypertrophy, heart weight index, through inhibition of phosphorylation of protein kinase B and cyclic adenosine monophosphate response element binding protein.
4. 4. The use according to claim 1, wherein the product is any one of a pharmaceutical product, a food product and a health product.
5. 5. The use according to any one of claims 1 to 4, wherein the product is in the form of any one of powder, granules, tablets and capsules.
6. 6. The drug monomer for resisting myocardial hypertrophy is characterized by comprising effective dose of DYZ-9.
7. 7. The anti-myocardial hypertrophy drug monomer according to claim 6, wherein the dosage form of the drug monomer is any one of powder, granule, tablet and capsule.
8. 8. A pharmaceutical composition for resisting myocardial hypertrophy, which is characterized by comprising an effective amount of DYZ-9 or pharmaceutically acceptable salts and hydrates thereof and pharmaceutically acceptable carriers, auxiliary agents or a composition thereof.
9. 9. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition is in the form of any one of powder, granule, tablet and capsule.

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