WO2023005002A1 - Use of sildenafil citrate in preparation of medicine for preventing and/or treating pulmonary arterial hypertension - Google Patents

Abstract

The present invention provides a use of sildenafil citrate in the preparation of a medicine used for preventing and/or treating pulmonary arterial hypertension. Sildenafil citrate may achieve cardiopulmonary protection for patients that have pulmonary arterial hypertension by means of inhibiting right ventricular remodeling, reducing cardiac hypertrophy, lowering a cardiac index, inhibiting pulmonary edema, inhibiting the thickening of pulmonary artery walls of different diameters, and lowering a pulmonary index. Sildenafil citrate also has a therapeutic effect on pulmonary arterial hypertension, which may improve the survival rate of pulmonary arterial hypertension model animals and increase the body weight of model animals. The therapeutic effect thereof is achieved by means of inhibiting the activity of PDE5 to adjust the level of cGMP in a PDE5/cGMP/PKG pathway.

Description

Use of edenafil citrate in the preparation of medicaments for preventing and or treating pulmonary hypertension technical field

The invention belongs to the technical field of new pharmacological effects of ardenafil citrate, and in particular relates to the use of ardenafil citrate in the preparation of medicines for preventing and or treating pulmonary hypertension.

Background technique

Pulmonary arterial hypertension (PAH) is a group of idiopathic pulmonary hemodynamic abnormalities caused by certain diseases, mainly manifested as increased pulmonary vascular resistance, pulmonary vascular remodeling, and in situ thrombosis, which can eventually lead to right ventricular hypertrophy , and progressively develop into right heart failure until death, which is a very serious chronic pulmonary circulation disease and a key link of cor pulmonale.

The prevention and treatment of pulmonary arterial hypertension is a major problem that needs to be studied and solved urgently. So far, there is still a lack of ideal drugs, and there is still a long-term problem of drug resistance to targeted drugs in clinical practice. It is necessary to replace drugs or combined therapy in a timely manner, so as to bring great benefits to the treatment of pulmonary arterial hypertension and prognosis. Come to a serious challenge. Pulmonary arterial hypertension is characterized by pulmonary arteriolar vascular remodeling and pulmonary artery vascular smooth muscle proliferation, manifested by increased pulmonary circulation pressure and resistance, increased right heart load, right heart dysfunction, and reduced pulmonary blood flow, resulting in a series of clinical manifestations ; During the course of the disease, pulmonary hypertension often develops progressively. In particular, right heart failure caused by enhanced pulmonary artery vascular tension is a disease that seriously threatens human life and health. Aiming at the main pathological basis in the pathogenesis of pulmonary arterial hypertension—pulmonary vascular remodeling, by studying the interaction and synergistic effect of multiple pathways, it can comprehensively and systematically explain the new mechanism of pulmonary arterial hypertension, and provide new ideas and new ideas for the prevention and treatment of pulmonary arterial hypertension. direction.

When pulmonary hypertension occurs, the patient will show symptoms such as dyspnea, palpitations, chest pain, hemoptysis, and syncope. In the late stage, hypoxia, right heart hypertrophy or even right heart failure will appear, and death will occur in severe cases. At present, bosentan is the first-line drug for the clinical treatment of pulmonary hypertension, but its therapeutic effect is not satisfactory. It is a very important and urgent work to develop new drugs that can effectively treat pulmonary hypertension.

In view of this, the present invention is proposed.

Contents of the invention

The technical problem to be solved in the present invention provides a new application of edenafil citrate in the preparation of drugs for the prevention and/or treatment of pulmonary hypertension. Male PD rats are used to establish an animal pulmonary hypertension model, which aims at preventing and treating pulmonary hypertension respectively. The dosing regimen was implemented in two directions, and the body weight and death of the animals were recorded during the dosing process. At the end of the experiment, the animal's electrocardiogram, right ventricular systolic pressure, and pulmonary artery pressure were detected; disease markers in serum were detected; organ indexes and cGMP content in lung tissue were measured, and pathological changes of pulmonary blood vessels were observed and judged. Preventive and/or therapeutic effects of edenafil citrate on pulmonary arterial hypertension.

The edenafil citrate is produced by heating and refluxing ardenafil and citric acid in absolute ethanol. The chemical name of edenafil citrate is: 1-[3-(6,7-dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo[4,3- d] pyrimidin-5-yl)-4-ethoxybenzenesulfonyl]-cis-3,5-dimethylpiperazine citrate, the molecular weight is 680.74, and the molecular formula is C ₂₃ H ₃₂ N ₆ O ₄ S·C ₆ H ₈ O ₇ has the following structure:

The present invention realizes the purpose of the invention through the following technical solutions:

The invention provides the use of edenafil citrate in the preparation of medicaments for preventing and/or treating pulmonary hypertension.

Specifically, its preventive effect is reflected in:

Aldenafil citrate protects the heart in the prevention of pulmonary hypertension by inhibiting right ventricular remodeling, reducing right ventricular hypertrophy, and reducing cardiac index.

Furthermore, edenafil citrate prevents pulmonary hypertension by inhibiting pulmonary edema and reducing lung index.

Furthermore, edenafil citrate prevents pulmonary hypertension by inhibiting the thickening of the walls of pulmonary arteries with different diameters.

Furthermore, edenafil citrate inhibited the cardiac injury caused by right ventricular remodeling by reducing the NT-proBNP content in lung tissue and serum.

Its therapeutic effects are reflected in:

The therapeutic effect of edenafil citrate on pulmonary hypertension includes improving the survival rate of pulmonary hypertension model animals and increasing their body weight.

Further, edenafil citrate regulates the level of cGMP in the PDE5/cGMP/PKG pathway by inhibiting the activity of PDE5, thereby realizing the treatment of pulmonary arterial hypertension.

In the above scheme, the PDE5/cGMP/PKG pathway is regulated by PDE5. Nitric oxide (NO) activates water-soluble guanylate cyclase (sGC) to catalyze the conversion of guanosine triphosphate (GTP) into the second messenger cyclic guanosine monophosphate ( cGMP), and the corresponding cGMP protein kinase (PKG) and the channel of the physiological process controlled by the cyclic nucleotide gate. These channels directly regulate smooth muscle motility, neurotransmission, regulation of blood-vesseltone, and immune response. The main function of PDE5 inhibitors is to accumulate intracellular cGMP by inhibiting the hydrolysis of cGMP by PDE5.

A further solution of the present invention is: the adult single application dose of edenafil citrate for preventing and treating pulmonary hypertension is 1-100 mg/kg, preferably 2.5-10 mg/kg.

The determination of the adult single dose in the above scheme is based on the calculation method based on the body surface area of humans and animals in pharmacological experiments. Specifically, the dosage formula for converting the clinical equivalent dose from animals to adults is: rat dose/human dose=rat specific surface area/human specific surface area, which can be obtained after adjustment: human dose=rat dose/( Rat specific surface area/human specific surface area), generally speaking, the average body surface area of a 60kg adult is 1.55m ² , the body surface area of a 200g rat is about 305cm ² , and the human specific surface area = 1.55/60 = 0.026, Rat specific surface area = 305/200/10 = 0.152. In the embodiment of the present invention, the preferred range of the dose for rats is 15-60 mg/kg, and the preferred range of single application dose for adults is 2.5-10 mg/kg through the conversion of the above formula.

A further scheme of the present invention is: the medicine for preventing and/or pulmonary hypertension comprises edenafil citrate compound and a pharmaceutically acceptable carrier or adjuvant, and its dosage form is selected from: solution, suspension, emulsion, In the form of pills, capsules, powders, controlled release, sustained release formulations and microsomal delivery systems.

A further solution of the present invention is: the drug for preventing and/or pulmonary hypertension is in an oral dosage form.

After adopting the above technical solution, the present invention has the following beneficial effects compared with the prior art:

Aldenafil citrate prevents cardiopulmonary protection in pulmonary hypertension by inhibiting right ventricular remodeling, reducing cardiac hypertrophy, reducing cardiac index, inhibiting pulmonary edema, inhibiting pulmonary artery wall thickening of different diameters, and reducing pulmonary index; It also has a therapeutic effect on pulmonary hypertension, which can improve the survival rate of pulmonary hypertension model animals and increase the weight of model animals. Its therapeutic effect is achieved by inhibiting the activity of PDE5 and adjusting the level of cGMP in the PDE5/cGMP/PKG pathway. .

The specific implementation manners of the present invention will be further described in detail below in conjunction with the accompanying drawings.

Description of drawings

The accompanying drawings, as a part of the present invention, are used to provide a further understanding of the present invention, and the schematic embodiments of the present invention and their descriptions are used to explain the present invention, but do not constitute improper limitations to the present invention. Apparently, the drawings in the following description are only some embodiments, and those skilled in the art can also obtain other drawings according to these drawings without creative efforts. In the attached picture:

Fig. 1 is that in the embodiment of the present invention 3, the electrocardiogram S wave deepening situation of the rat at the end point of the preventive effect experiment is schematic;

Fig. 2 is a schematic representation of right ventricular systolic pressure and right ventricular remodeling at the end point of the preventive effect experiment in rats in Example 3 of the present invention;

Fig. 3 is a schematic diagram of the systemic circulation of rats at the end point of the preventive effect experiment in Example 4 of the present invention;

Fig. 4 is a schematic diagram of the heart and lung index situation of rats at the end point of the preventive effect experiment in Example 5 of the present invention;

Figure 5 is a schematic representation of the situation of NT-proBNP in the serum and lung tissue of rats at the end point of the preventive effect experiment in Example 6 of the present invention;

Fig. 6 is the schematic representation of the pulmonary vascular structure situation of rats at the end point of the preventive effect experiment in Example 7 of the present invention;

Figure 7 is a schematic diagram of the body weight of the rats at the end of the therapeutic effect experiment in Example 9 of the present invention;

Figure 8 is a schematic diagram of the right ventricular remodeling of rats at the end of the therapeutic effect experiment in Example 11 of the present invention;

Fig. 9 is a schematic representation of the cGMP content in the lung tissue of rats at the end point of the treatment effect experiment in Example 12 of the present invention.

It should be noted that these drawings and text descriptions are not intended to limit the concept scope of the present invention in any way, but illustrate the concept of the present invention for those skilled in the art by referring to specific embodiments.

Detailed ways

In order to make the purpose, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. The following embodiments are used to illustrate the present invention , but not to limit the scope of the present invention.

Example 1

In this example, from the perspective of preventing pulmonary hypertension, a model of pulmonary hypertension in rats was established, and a preventive drug regimen was implemented.

Example Animals 70 SPF grade SD rats were selected, with a body weight of 200-220 g, and were randomly divided into 7 groups, with 10 rats in each group. The above 7 groups were respectively set as normal control group, pulmonary hypertension model group, and positive drug sildenafil group (30mg/kg, oral administration, once a day), positive drug bosentan group (100mg/kg, oral administration, once a day), edenafil citrate low dose group (15mg/kg, oral administration , once a day), edenafil citrate medium dose group (30mg/kg, oral administration, once a day) and edenafil citrate high dose group (60mg/kg, oral administration, once a day) . At the beginning of the experiment, in addition to the normal control group given the same amount of normal saline, the other animals were given 50 mg/kg monocrotaline subcutaneously at one time, and the rat pulmonary hypertension model was established in other groups, and the continuous administration was started on the day after the model was established as day1. For 21 days, animal body weight was monitored and mortality was recorded every 4 days, once a day.

After the last administration on Day 21, the animals to be tested were fasted overnight, and their body weight was weighed and recorded the next day. The electrocardiogram, right ventricular systolic pressure, and systemic circulation pressure were detected at the end point of the experiment. After the experiment, blood was taken from the abdominal aorta for the detection of various indicators, and the heart, lung, liver and other organs were weighed to calculate the organ index. Parts of the heart, lungs, and pulmonary arteries were used for pathological examination.

The organ detection specifically includes:

(1) Open the thorax of the rat at the end of the experiment, take out the lungs, heart, and liver, rinse in normal saline to remove congestion, use filter paper to dry up the residual liquid, weigh it, and compare it with the body weight of the day to obtain the organ index;

(2) The lungs were perfused with normal saline, dried with filter paper, and the upper lobe of the left lung was taken, cut longitudinally close to the branch of the left pulmonary artery, leaving the outer lobe, fixed with 4% paraformaldehyde, embedded in paraffin and sectioned, then stained with Masson , observed pulmonary vascular remodeling and changes in alveolar structure, and counted the thickness of blood vessel walls with three different diameters (n=4).

(3) Take 0.2 g of frozen right lung tissue, add 1 mL of 1% PBS, grind and homogenate with a hand-held homogenizer, centrifuge at 1500 g for 10 min at 4°C, and process according to the kit instructions for NT-proBNP detection.

(4) Cut off the entire ventricle along the lower edge of the atrial appendage, then cut off the right ventricle along the edge of the interventricular septum, weigh it, and calculate the ratio of the right ventricle (RV) to the sum of the left ventricle and interventricular septum (L+S), and calculate [RV/ (L+S)], namely the Fulton index; the ratio of right ventricle (RV) and body weight is calculated to calculate RV/BW, namely the right ventricle index, both of which can reflect the remodeling situation and degree of hypertrophy of the right ventricle.

For the selection of administration concentration in the present embodiment, it should be noted that when the concentration of aldenafil citrate is too high (such as 100mg/kg), it will have a certain impact on experimental animals, so the present invention is in the selected high-dose group. When it is lower than the above-mentioned concentration of 100mg/kg.

Example 2

This example is to study the effect of edenafil citrate on the survival rate of pulmonary hypertension model rats from the perspective of preventing pulmonary hypertension.

At the end of the experiment, the death of animals was counted. The results are shown in Table 1, 2 animals died in the model group, and no animals died in the blank group and each administration group, indicating that the administration of Aldenafil citrate can delay the development of pulmonary hypertension and play a preventive role. while reducing mortality.

Rat survival rate at the end point of the preventive effect experiment in table 1

Example 3

This example is to study the influence of edenafil citrate on cardiac function, right ventricular systolic pressure and right ventricular remodeling of pulmonary hypertension model rats from the perspective of preventing pulmonary hypertension.

At the end of the experiment, the rats were anesthetized with chloral hydrate, their limbs and heads were fixed on the mouse board, and the heating plate was turned on to maintain the animal's body temperature. Needle-shaped electrodes were inserted subcutaneously in the front and rear limbs, and the changes in lead II ECG were observed, recorded for 3 minutes, and the changes in indicators related to right ventricular dysfunction were observed, including: P wave, QRS wave, and S wave, and the deepening of the S wave was counted.

On the basis of the existing neck incision in the rat, the right neck muscle was further exposed, the tissue was bluntly dissected, and the external jugular vein with a length of about 1 cm was freed, two sutures were embedded, the distal end was ligated, and the proximal end was scissored. With a V”-shaped incision, slowly push the catheter filled with heparin sodium solution into the superior vena cava and enter the right atrium. When the right ventricle relaxes, pass the catheter through the tricuspid valve and enter the right ventricle. After the blood pressure is stable, record the right ventricular systolic pressure (RVSP )3min.

As shown in Figure 1, the electrocardiographic S wave of rats with pulmonary hypertension deepened, suggesting that the right heart function of the animals was damaged. High-dose edenafil has a tendency to inhibit the deepening of S wave, indicating that it improves cardiac function to a certain extent.

As shown in Figure 2, the systolic pressure of the right ventricle of the rats with pulmonary arterial hypertension in each group was detected, and the systolic pressure of the right ventricle of the model group was significantly higher than that of the blank group. There is a downward trend. Compared with normal animals, the Fulton index of the model group is right heart hypertrophy index RV/(L+S) (as shown in Figure 2 B), right heart index RV/BW (as shown in Figure 2 C) and ventricular index (as shown in D in Figure 2) were all significantly increased, and at the same time, each dose group edenafil could significantly reduce the right heart hypertrophy degree of pulmonary hypertension rats, and showed a better dose dependence, indicating that Aldenafil citrate has a strong inhibitory effect on right heart injury in pulmonary arterial hypertension rats.

Example 4

This example is to study the effect of edenafil citrate on the systemic circulation of pulmonary hypertension model rats from the perspective of preventing pulmonary hypertension.

At the end of the experiment, the left carotid artery was intubated, a median incision was made in the rat's neck, the muscle nerves were separated, the blood vessels were stripped out, 3 sutures were embedded, the distal end was ligated with a thread, and the proximal end was temporarily blocked with a thread. Make a "V"-shaped incision, insert a polyethylene plastic tube (PE50) pre-filled with heparin sodium solution (100U/mL), connect the other end of the polyethylene plastic tube to the BL420 biological function experiment instrument, and fix the tube and blood vessel with a thread. Loosen the proximal line, restore blood flow, and record aortic pressure, including systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) after the blood pressure is stabilized.

Systemic circulation in rats with pulmonary hypertension was monitored. One is to observe whether the drug has an effect on the systemic circulation pressure and heart rate; the other is to observe whether the drug has an effect on the systemic circulation pressure and heart rate; the other is that when the pulmonary hypertension develops to a certain extent, the heart function is damaged and the systemic circulation pressure changes. Severity of pulmonary hypertension.

The systemic circulation of animals in each group was detected by catheterization of the left carotid artery. As shown in Figure 3, the systolic blood pressure and diastolic blood pressure of the model group were significantly reduced (#P<0.05, ##P<0.01), indicating that the animal's heart The function has been seriously damaged, and this result can also explain the lower-than-expected RVSP of the model animal from the side. In addition, although the systemic circulation pressure and heart rate of the animals in each administration group were not significantly different from those in the model group, they were close to those in the normal control group, indicating that the administration of drugs can improve the heart function of the animals.

Example 5

This example is to study the influence of edenafil citrate on the indexes of various organs in rats with pulmonary arterial hypertension from the perspective of preventing pulmonary arterial hypertension.

Various organs extracted in the organ detection step in Example 1 were taken for index determination. After continuous administration for 21 days, the organ indexes of animals in the model group and each administration group changed. The results showed that as shown in Figure 4, during the development of pulmonary hypertension, the heart index and lung index of the model animals were significantly increased (###P<0.001), suggesting that the heart and lungs of the rats were damaged. Giving the positive drug and three doses of low, middle and high doses of edenafil can significantly reduce the cardiac index, indicating that ardenafil has a protective effect on the heart of animals; and low doses of edenafil can significantly reduce pulmonary edema, suggesting that low doses of edenafil can significantly reduce the cardiac index Dose edenafil may have an advantage in protecting the lungs.

Example 6

This example is to study the effect of edenafil citrate on NT-proBNP in serum and lung tissue of rats with pulmonary hypertension from the perspective of preventing pulmonary hypertension.

Use the ELISA method to detect the content of NT-proBNP in serum and lung tissue. Prepare various reagents according to the kit instructions, equilibrate at room temperature for 30 minutes, and set blank wells, standard sample wells, and test sample wells for each well. Add 100 μL of standard substance or sample to be tested, shake well, cover the plate, incubate at 37°C for 2 hours, discard the liquid, and shake dry; add 100 μL of biotin-labeled antibody working solution to each well, cover the plate, and incubate at 37°C 1h, discard the liquid in the well, spin dry, add 200μL of detergent to each well, wash 3 times for 2min/time; add 100μL of the working solution of horseradish peroxidase-labeled avidin to each well, and cover the plate , incubate at 37°C for 1 hour, discard the liquid in the well, spin dry, and wash the plate 5 times according to the above operation method; add 90 μL of substrate reaction solution to each well, develop color at 37°C for 15-30 minutes in the dark, add 50 μL of stop reaction solution, and stop reaction. Within 5 minutes, the absorbance was detected at 450nm with a microplate reader.

The level of NT-proBNP in serum and lung tissue can be used as an important indicator of heart failure in patients. As shown in Figure 5 by the serum sample detection results, the NT-proBNP content in the serum of the rats in the model group increased significantly, and the NT-proBNP content in the serum of each administration group decreased significantly, indicating that ardenafil has a significant effect on the damage of the heart. significant inhibition. The NT-proBNP in the lung tissue of rats in the model group also tended to increase, but compared with the serum, there was no significant difference in the change of NT-proBNP content in the tissue.

Example 7

This example is to study the effect of edenafil citrate on the pulmonary vascular structure of pulmonary hypertension model rats from the perspective of preventing pulmonary hypertension.

Get the different diameters of pulmonary artery blood vessels extracted in the organ detection step in Example 1, apply Masson staining, can observe the shape of lung alveoli and blood vessels, count the blood vessel wall thickness of three kinds of diameter ranges in the PAH rat lung tissue, The remodeling trends of blood vessels with different diameters were roughly the same, as shown in Figure 6.

The alveolar morphological structure of the rats in the PAH model group was damaged, and the walls of blood vessels in three diameter ranges were thicker than those of the same diameter in the control group. Compared with the PAH model group, the positive drug bosentan group can significantly reduce the thickness of the blood vessel wall in the lung tissue with a blood vessel diameter greater than 100 μm. In addition, bosentan also has a tendency to reduce the thickness of blood vessel wall in PAH rats with blood vessel diameter less than 100 μm. The positive drug, sildenafil, has a tendency to reduce the blood vessel walls in three diameter ranges in the lungs of PAH model animals.

And three concentrations of biological edenafil citrate (15mg/kg, 30mg/kg, 60mg/kg) can significantly reduce the blood vessel wall thickness of the three diameter ranges of the blood vessel wall thickness in the PAH rat lung tissue and alleviate disease state. Among them, the middle dose group (30 mg/kg) has the best effect on the blood vessels in the lung tissue with a blood vessel diameter of 50 to 100 μm, while the high dose group has the strongest inhibitory effect on the thickening of the pulmonary blood vessel walls in the rest of the diameter range.

Example 8

In this example, from the perspective of treating pulmonary hypertension, a model of pulmonary hypertension in rats was established, and a therapeutic dosing regimen was implemented.

Example Animals 70 SPF grade SD rats were selected, with a body weight of 200-220 g, and were randomly divided into 7 groups, with 10 rats in each group. The above 7 groups were respectively set as normal control group, pulmonary hypertension model group, and positive drug sildenafil group (30mg/kg, oral administration, once a day), positive drug bosentan group (100mg/kg, oral administration, once a day), edenafil citrate low dose group (15mg/kg, oral administration , once a day), edenafil citrate medium dose group (30mg/kg, oral administration, once a day) and edenafil citrate high dose group (60mg/kg, oral administration, once a day) . At the beginning of the experiment, in addition to the normal control group given the same amount of normal saline, the other animals were given 60 mg/kg monocrotaline by subcutaneous injection at one time, and the rat pulmonary hypertension model was established with other groups. The PAH disease of the animals had disappeared on the 14th day after the model was established. When the stability was formed, the model animals were regrouped on day 14 and continued to be administered until the mortality rate of the model group reached 70%.

When the mortality rate of the model group reached 70%, after the last administration, the animals to be tested were fasted overnight, and their body weight was weighed and recorded the next day. The experimental endpoint detection indicators are as follows:

(1) Take out the lung and wash it with normal saline, blot dry with filter paper, take 0.2g of frozen right lung tissue, add 1mL PBS, grind and homogenate with an automatic grinder, centrifuge at 1500g for 10min at 4°C, and process according to the instructions , for cGMP testing.

(2) Cut off the entire ventricle along the lower edge of the atrial appendage, then cut off the right ventricle along the edge of the interventricular septum, weigh it, and calculate the ratio of the right ventricle (R) to the sum of the left ventricle and interventricular septum (L+S), and calculate [R/ (L+S)], the ratio of right ventricle (R) to body weight R/BW, calculate the right ventricle index, and calculate the degree of hypertrophy of the right ventricle.

Example 9

This example is to study the effect of edenafil citrate on the body weight of pulmonary hypertension model rats from the perspective of treating pulmonary hypertension.

After 14 days of modeling, the model animals were regrouped, and the administration was started, and the body weight was recorded at the same time. As shown in Figure 7, the body weight difference between the normal group and the model group was very obvious after 14 days of modeling, and the body weight of the modeled animals was significantly lower. At the end of the experiment, the middle dose of edenafil citrate has a tendency to improve the body weight of the animals.

Example 10

This example is to study the influence of edenafil citrate on the survival rate of pulmonary hypertension model rats from the perspective of treating pulmonary hypertension.

As shown in Table 2, after 14 days of single-administration of MCT for modeling, continuous administration for 16 days began, and only 40% of the rats in the model group survived. The survival rates of the mice were 40% and 60% respectively. Bosentan did not improve the survival rate of the animals, but it had a certain effect of delaying the death. Sildenafil and aldenafil citrate in each dose group showed the effect of improving the survival rate of animals. However, high-dose edenafil citrate can significantly improve the survival rate of pulmonary hypertension model rats to 70%, much higher than positive drugs.

Rat survival rate at the end point of the treatment effect experiment in table 2

Example 11

This example is to study the effect of edenafil citrate on the right ventricle remodeling of pulmonary hypertension model rats from the perspective of treating pulmonary hypertension.

At the end of the experiment, the heart weight of the rat was weighed and compared with the corresponding body weight to obtain the cardiac index, ventricular index, and right ventricular index, and to calculate the Fulton index. As shown in Figure 8, the right heart of the animals in the model group was significantly hypertrophic, and high doses of edenafil citrate had significant effects on cardiac index (shown in Figure 8 in A), ventricular index (shown in Figure 8 in B), right ventricular The index (shown as C in Figure 8) has a certain improvement trend.

Example 12

This example is to study the effect of edenafil citrate on cGMP in lung tissue of rats with pulmonary hypertension from the perspective of treating pulmonary hypertension.

As shown in Figure 9, the cGMP content in the lung tissue of the rats in the model group decreased slightly, and the cGMP content in the lung tissue of the animals in each dose group of edenafil citrate had a certain increase, reflecting that edenafil citrate By inhibiting the activity of PDE5, the level of cGMP in the PDE5/cGMP/PKG pathway can be adjusted to achieve the therapeutic effect on pulmonary hypertension.

Comprehensive analysis embodiment 1～12

Wherein according to the content of embodiment 1～7, it can be seen that given different doses of the test drug edenafil citrate, no death occurred in the animals, while the death rate of the model group animals was 20%, which shows that edenafil citrate Non-true can delay the occurrence and development of PAH, and improve the survival time and survival rate of model animals.

The ECG and right ventricular systolic pressure were detected, and the results showed that edenafil citrate had no significant effect on reducing right ventricular systolic pressure (ie, pulmonary artery pressure), but inhibited the remodeling phenomenon of the right ventricle very significantly, indicating that edenafil citrate Acid aldenafil is more focused on the protection of the heart in the treatment of pulmonary hypertension.

In terms of organ indexes, edenafil citrate can significantly reduce cardiac hypertrophy and pulmonary edema, which indirectly reflects its cardiopulmonary protective effect.

In terms of pulmonary artery remodeling, the observation of pathological tissue sections showed that the PAH model group had severe pulmonary artery remodeling, pulmonary interstitial edema, and alveolar structure damage. Statistics on the thickness of blood vessel walls in three diameter ranges (<50 μm, 50-100 μm, >100 μm) found that the pulmonary artery wall of rats with pulmonary arterial hypertension was thickened, and edenafil citrate could significantly inhibit pulmonary vascular weight. Structural phenomenon, and the curative effect is better than positive drugs.

The disease marker NT-proBNP is currently one of the auxiliary indicators for the diagnosis of PAH. From the experimental results, the content of NT-proBNP in the serum of the model animals was significantly increased, and each dose of edenafil citrate had no effect on the effect of the disease markers. The increase has a significant decrease effect, which further proves the inhibitory effect of the drug on the right heart injury.

It can be seen that edenafil citrate can significantly inhibit the phenomenon of pulmonary artery vascular remodeling, and has a prominent inhibitory effect on the structure and function of the right ventricle. The protective effect of remodeling inhibitors on the heart and blood vessels is that it delays the development of the disease , An important reason to improve the survival rate of animals, and thereby play a good preventive effect on pulmonary hypertension.

Continue to analyze the content of embodiment 8～12, when the death rate of model group reaches 70% (being administered 16/14 days), experiment terminates, and positive drug sildenafil shows the effect of raising animal survival rate preferably now. effect. Each dose of edenafil citrate has a significant effect on improving the survival rate, and the effect of high dose is more prominent and stronger than that of the positive drug sildenafil. In addition, the middle dose of edenafil citrate can increase the body weight of the rats, which may improve the state of the rats to a certain extent.

High doses of edenafil citrate can improve cardiac index, ventricular index, and right ventricular index, while the cGMP content in the lung tissue of animals in each dose group of edenafil citrate has a certain increase , showing that edenafil citrate regulates the level of cGMP in the PDE5/cGMP/PKG pathway by inhibiting the activity of PDE5 to achieve the therapeutic effect on pulmonary hypertension.

The above description is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as above with preferred embodiments, it is not intended to limit the present invention. Anyone familiar with the technology of this patent Without departing from the scope of the technical solution of the present invention, personnel can use the technical content of the above prompts to make some changes or modify them into equivalent embodiments with equivalent changes. In essence, any simple modifications, equivalent changes and modifications made to the above embodiments still fall within the scope of the solutions of the present invention.

Claims (10)

Use of edenafil citrate in preparing a medicament for preventing and/or treating pulmonary hypertension. The use according to claim 1, characterized in that edenafil citrate protects the heart in the prevention of pulmonary hypertension by inhibiting right ventricular remodeling, reducing right ventricular hypertrophy, and reducing cardiac index. The use according to claim 1, characterized in that edenafil citrate prevents pulmonary hypertension by inhibiting pulmonary edema and reducing pulmonary index. The use according to claim 1, characterized in that the edenafil citrate prevents pulmonary hypertension by inhibiting the thickening of the walls of pulmonary arteries with different diameters. The use according to claim 2, characterized in that edenafil citrate inhibits cardiac damage caused by right ventricular remodeling by reducing the NT-proBNP content in lung tissue and serum. The use according to claim 1, characterized in that the therapeutic effect of edenafil citrate on pulmonary hypertension includes improving the survival rate of pulmonary hypertension model animals and increasing their body weight. The use according to claim 6, characterized in that edenafil citrate achieves the treatment of pulmonary arterial hypertension by inhibiting the activity of PDE5 to regulate the level of cGMP in the PDE5/cGMP/PKG pathway. The use according to any one of claims 1-7, characterized in that the adult single application dose of edenafil citrate for preventing and treating pulmonary hypertension is 1-100 mg/kg, preferably 2.5-10 mg/kg . The use according to claim 8, characterized in that, the medicine for preventing and/or pulmonary hypertension comprises a preparation made of edenafil citrate compound and a pharmaceutically acceptable carrier or adjuvant, and the dosage form of the preparation is selected from From: solutions, suspensions, emulsions, pills, capsules, powders, controlled release, sustained release formulations and microsomal drug delivery systems. The use according to claim 9, characterized in that the medicament for preventing and/or pulmonary hypertension is an oral dosage form.

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