CN114762692A

CN114762692A - Application of Aidinafei citrate in preparing medicine for preventing and/or treating pulmonary hypertension

Info

Publication number: CN114762692A
Application number: CN202110856061.8A
Authority: CN
Inventors: 杨磊; 杜冠华; 袁天翊; 赵晓悦; 马国栋; 燕柳艳; 宋俊科
Original assignee: Yuekang Pharmaceutical Group Co ltd
Current assignee: Yuekang Pharmaceutical Group Co ltd
Priority date: 2021-07-28
Filing date: 2021-07-28
Publication date: 2022-07-19
Also published as: WO2023005002A1

Abstract

The invention discloses an application of Aidinafil citrate in preparing a medicine for preventing and/or treating pulmonary hypertension. The invention has the advantages that: the citric acid alidenafil can reduce cardiac hypertrophy, reduce cardiac index, inhibit pulmonary edema, inhibit the thickening of the wall of pulmonary artery blood vessels with different diameters and reduce the pulmonary index by inhibiting the reconstruction of the right ventricle, thereby realizing the cardiopulmonary protection of patients with pulmonary hypertension; the compound also has a therapeutic effect on pulmonary hypertension, can improve the survival rate of a pulmonary hypertension model animal and increase the body weight of the model animal, and the therapeutic effect is realized by regulating the cGMP level in a PDE5/cGMP/PKG pathway by inhibiting the activity of PDE 5. Therefore, the citric acid Aidi has good application and development prospect in preventing and/or treating pulmonary hypertension and complications thereof.

Description

Application of Aidinafei citrate in preparing medicine for preventing and/or treating pulmonary hypertension

Technical Field

The invention belongs to the technical field of new pharmacological effects of Aidinafei citrate, and particularly relates to application of Aidinafei citrate in preparing a medicine for preventing and/or treating pulmonary hypertension.

Background

Pulmonary Arterial Hypertension (PAH) is a group of specific diseases with pulmonary hemodynamic abnormalities caused by certain diseases, mainly manifested by increased pulmonary vascular resistance, pulmonary vascular remodeling and in-situ thrombosis, which can finally lead to right ventricular hypertrophy and progressive development into right heart failure until death, and is a very serious chronic pulmonary circulation disease and also a key link of pulmonary heart disease.

The prevention and treatment of pulmonary hypertension is a major problem to be researched and solved urgently at present, an ideal medicament is still lacked so far, the problem of targeted medicament resistance exists in clinical practice for a long time, and medicaments or combined treatment must be replaced timely, so that the treatment and the prognosis rehabilitation of pulmonary hypertension are provided with serious challenges. Pulmonary hypertension is characterized by the pathological features of pulmonary arteriolar vascular remodeling and pulmonary arterial vascular smooth muscle proliferation, and is characterized by increased pulmonary circulation pressure and resistance, increased right heart load, right heart dysfunction and reduced pulmonary blood flow, thereby causing a series of clinical manifestations; pulmonary hypertension often develops progressively during the course of the disease. In particular to right heart failure caused by strengthening pulmonary artery vascular tension, which is a disease seriously threatening the life and health of human beings. Aiming at the main pathological basis in the pulmonary hypertension attack process, namely pulmonary vessel reconstruction, by researching the interaction and synergistic effect of multiple channels, the novel mechanism of pulmonary hypertension generation can be comprehensively and systematically explained, and a novel thought and a novel direction are provided for preventing and treating pulmonary hypertension.

When pulmonary hypertension occurs, patients show symptoms such as dyspnea, palpitation, chest pain, hemoptysis, syncope and the like, and in the late stage, the symptoms such as anoxia, right heart hypertrophy and even right heart failure occur, and death occurs in severe cases. At present, the first-line medicament for clinically treating pulmonary hypertension is bosentan, but the treatment effect is not satisfactory. It is a very important and urgent task to develop new drugs that can effectively treat pulmonary hypertension.

In view of the above, the present invention is particularly proposed.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a new application of Aidinafil citrate in preparing a medicament for preventing and/or treating pulmonary hypertension. Detecting the electrocardiogram, right ventricular systolic pressure and pulmonary artery pressure of the animal at the experimental end point; detecting disease markers in the serum; measuring organ index and cGMP content in lung tissue, observing pathological change of lung blood vessel, and judging the preventing and/or treating effect of Aidinafil citrate on pulmonary hypertension.

The citric acid alidenafil is generated by heating and refluxing the alidenafil and citric acid in absolute ethyl alcohol. The chemical name of the citric acid alidenafil is: 1- [ 3- (6, 7-dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo [4, 3-d)]Pyrimidine-5-yl) -4-ethoxy benzenesulfonyl ] -cis-3, 5-dimethyl piperazine citrate with molecular weight of 680.74 and molecular formula C₂₃H₃₂N₆O₄S·C₆H₈O₇The structure is as follows:

the invention realizes the purpose through the following technical scheme:

the invention provides application of Aidinafei citrate in preparing a medicament for preventing and/or treating pulmonary hypertension.

Specifically, the prevention effect is shown in the following steps:

the citric acid alidenafil can reduce the right ventricular hypertrophy and the cardiac index by inhibiting the right ventricular remodeling, thereby realizing the protection of the heart in the prevention of pulmonary hypertension.

Furthermore, the citric acid alidenafil can prevent pulmonary hypertension by inhibiting pulmonary edema and reducing pulmonary index.

Furthermore, the citric acid alidenafil can prevent pulmonary hypertension by inhibiting the thickening of the wall of the pulmonary artery blood vessel with different diameters.

Furthermore, the citric acid alidenafil can inhibit the heart damage caused by the reconstruction of the right ventricle by reducing the content of NT-proBNP in lung tissues and serum.

The treatment effect is shown in:

the therapeutic effect of the citric acid aildenafil on pulmonary hypertension includes improving the survival rate of the pulmonary hypertension model animal and increasing the weight of the pulmonary hypertension model animal.

Furthermore, the citric acid alidenafil can regulate the cGMP level in the PDE5/cGMP/PKG pathway by inhibiting the activity of PDE5, thereby realizing the treatment of pulmonary hypertension.

In the above scheme, the PDE5/cGMP/PKG pathway is the pathway through which PDE5 regulates physiological processes that are controlled by Nitric Oxide (NO) -activated water-soluble guanylate cyclase (sGC) to catalyze the conversion of Guanosine Triphosphate (GTP) to the second messenger cyclic guanosine monophosphate (cGMP), and the corresponding cGMP Protein Kinase (PKG) and cyclic nucleotide gates. These pathways directly regulate smooth muscle motility, neurotransmission (neurotransmission), vascular tension (regulation of blood-vascular tone), and immune response (immune response). The main function of PDE5 inhibitors is to allow accumulation of intracellular cGMP by inhibiting hydrolysis of cGMP by PDE 5.

The further scheme of the invention is as follows: the single-use dosage of the Aidinafil citrate for preventing and treating pulmonary hypertension for adults is 1-100 mg/kg, preferably 2.5-10 mg/kg.

The single dosage of the human in the scheme is determined according to a body surface area calculation method of the human and the animal in a pharmacological experiment. Specifically, the formula of the dosage of the clinical equivalent dose converted from animal to adult is as follows: rat dose/human dose-rat specific surface area/human specific surface area, adjusted to give: human dose-rat dose/(rat specific surface area/human specific surface area), in generalThe average body surface area of an adult weighing 60kg was 1.55m²The body surface area of a 200g rat is about 305cm²The specific surface area of human is 1.55/60-0.026, and the specific surface area of rat is 305/200/10-0.152. In the embodiment of the invention, the optimal range of the rat dose is 15-60 mg/kg, and the optimal range of the adult single-use dose can be calculated to be 2.5-10 mg/kg through the conversion of the formula.

The further scheme of the invention is as follows: the medicine for preventing and/or treating pulmonary hypertension comprises an Aidenafil citrate compound and a pharmaceutically acceptable carrier or auxiliary material, and the dosage form of the medicine is selected from: solutions, suspensions, emulsions, pills, capsules, powders, controlled release, sustained release formulations and microsomal delivery systems.

The further scheme of the invention is as follows: the medicine for preventing and/or treating pulmonary hypertension is an oral dosage form.

After adopting the technical scheme, compared with the prior art, the invention has the following beneficial effects:

the citric acid alidenafil can be used for preventing the pulmonary hypertension from protecting the heart and lungs by inhibiting the reconstruction of the right ventricle, reducing the cardiac hypertrophy, reducing the cardiac index, inhibiting pulmonary edema, inhibiting the thickening of the wall of the pulmonary artery with different diameters and reducing the pulmonary index; it also has therapeutic effect on pulmonary hypertension, and can improve survival rate of pulmonary hypertension model animal and increase body weight of model animal, and the therapeutic effect is realized by inhibiting activity of PDE5 to regulate cGMP level in PDE5/cGMP/PKG pathway

The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention, 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 without limiting the invention to the right. It is obvious that the drawings in the following description are only some embodiments, and that for a person skilled in the art, other drawings can be derived from them without inventive effort. In the drawings:

FIG. 1 is a schematic diagram showing the deepening of the electrocardiogram S wave of the rat at the end point of the preventive effect experiment in example 3 of the present invention;

FIG. 2 is a graph showing the right ventricular systolic pressure and right ventricular remodeling of the rat in example 3 of the present invention at the experimental endpoint of the preventive effect;

FIG. 3 is a schematic representation of the systemic circulation of rats at the end of the prophylactic effect test in example 4 of the present invention;

FIG. 4 is a graph showing the index of heart and lung of the rat at the end point of the experiment of preventing effect in example 5 of the present invention;

FIG. 5 is a diagram showing NT-proBNP in serum and lung tissues of rats at the end point of the experiment on the prevention effect in example 6 of the present invention;

FIG. 6 is a diagram showing the pulmonary vascular structure of a rat in example 7 of the present invention at the end of the preventive effect test;

FIG. 7 is a schematic representation of the body weight of the rat at the end of the therapeutic effect experiment in example 9 of the present invention;

FIG. 8 is a graph showing the right ventricular remodeling in the therapeutic effect experiment end point of the rat in example 11 of the present invention;

FIG. 9 is a graph showing cGMP content in lung tissue at the end of the therapeutic effect experiment in rats in example 12 of the present invention.

It should be noted that the drawings and the description are not intended to limit the scope of the inventive concept in any way, but to illustrate it by a person skilled in the art with reference to specific embodiments.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and the following embodiments are used for illustrating the present invention and are not intended to limit the scope of the present invention.

Example 1

In this example, from the viewpoint of preventing pulmonary hypertension, a rat pulmonary hypertension model is established, and a prophylactic administration regimen is performed.

In the examples, 70 SPF-grade SD rats were selected, weighed 200 to 220g, and randomly divided into 7 groups of 10 animals each, and the 7 groups were set as a normal control group, a pulmonary hypertension model group, a positive drug sildenafil group (30mg/kg, oral administration, once a day), a positive drug bosentan group (100mg/kg, oral administration, once a day), a citric acid aildenafil low dose group (15mg/kg, oral administration, once a day), a citric acid aildenafil medium dose group (30mg/kg, oral administration, once a day), and a citric acid aildenafil high dose group (60mg/kg, oral administration, once a day). In addition to normal control group, other animals are given 50mg/kg of monocrotaline by one-time subcutaneous injection at the beginning of the experiment, and the other animals are established into a rat pulmonary artery hypertension model by other groups, and are continuously given for 21 days by day of molding as day1, the weight of the animals is monitored once a day and the death condition is recorded every 4 days.

After the last dose of Day21, the animals to be tested were fasted overnight, weighed the next Day and recorded. The end point of the experiment detects electrocardiogram, right ventricular systolic pressure and systemic circulation pressure. After the experiment is finished, the abdominal aorta is subjected to blood sampling for detecting each index, and visceral organs such as heart, lung, liver and the like are weighed to calculate visceral organ indexes. Part of heart, lung and pulmonary artery are used for pathological detection.

The organ detection specifically includes:

(1) opening the chest cavity of a rat at the end point of the experiment, taking out the lung, the heart and the liver, rinsing in normal saline, removing blood stasis, sucking the residual liquid by using filter paper, weighing, and comparing with the weight of the day to obtain the index of the viscera;

(2) the lung is washed by normal saline perfusion, water is absorbed by filter paper, the upper left lung lobe is taken, the upper left lung lobe is longitudinally cut close to the branch of the left pulmonary artery, the outer lung lobe is kept, the lung is fixed by 4 percent paraformaldehyde, the horse pine is stained after paraffin embedding and slicing, the pulmonary vascular remodeling condition and the pulmonary alveolus structural change are observed, and the thicknesses of three vascular walls with different diameters are counted (n is 4).

(3) 0.2g of frozen right lung tissue is taken, 1mL of 1% PBS is added, a hand-held homogenizer is used for grinding and homogenizing, centrifugation is carried out at 1500g for 10min at 4 ℃, and the processing operation is carried out according to the kit instruction for NT-proBNP detection.

(4) Cutting off the whole ventricle along the lower edge of the auricle, then cutting off the right ventricle along the edge of the ventricular septum, weighing, taking the ratio of the Right Ventricle (RV) to the sum of the left ventricle and the ventricular septum (L + S), and calculating [ RV/(L + S) ], namely a Fulton index; the ratio of Right Ventricle (RV) to body weight is calculated as RV/BW, the index of the right ventricle, which reflects the remodeling and hypertrophy of the right ventricle.

For the concentration selection in this example, it should be noted that the concentration of the citric acid alidenafil is too high (e.g. 100mg/kg), which has a certain effect on the experimental animals, so the concentration of the citric acid alidenafil in the selected high dose group is lower than the above 100 mg/kg.

Example 2

In the embodiment, from the perspective of preventing pulmonary hypertension, the influence of the Aidenafil citrate on the survival rate of the pulmonary hypertension modeled rats is studied.

At the end of the experiment, statistics were carried out on the death of the animals. The results are shown in table 1, 2 animals died in the model group, and no animal death occurred in the blank group and each administration group, which indicates that the administration of the citric acid Aidinafei can delay the disease development process of pulmonary hypertension, play a role in preventing and reducing the death rate.

Table 1 prevention effect survival rate of rats at the end of the experiment

Example 3

In this example, the influence of citric acid alidenafil on the cardiac function, right ventricular systolic pressure and right ventricular remodeling of pulmonary hypertension modeled rats is studied from the viewpoint of preventing pulmonary hypertension.

At the end of the experiment, rats were anesthetized with chloral hydrate, their extremities and head were mounted on rat plates, and the animal body temperature was maintained by opening the heating plate. Inserting needle electrodes into front and back limbs subcutaneously, observing II-lead electrocardiogram change, recording for 3min, and observing index change related to right ventricular dysfunction, including: p wave, QRS wave and S wave, and counting the deepening condition of the S wave.

On the basis of the existing neck incision of a rat, right side neck muscles are further exposed, tissues are separated in a blunt manner, external jugular veins with the length of about 1cm are dissociated, 2 threads are buried, the far end is ligated, the near end is cut into a V-shaped incision, a catheter filled with a heparin sodium solution is slowly pushed into the superior vena cava to enter the right atrium, when the right ventricle relaxes, the catheter passes through the tricuspid valve to enter the right ventricle, and after the blood pressure is stabilized, the Right Ventricular Systolic Pressure (RVSP) is recorded for 3 min.

As shown in figure 1, the cardiac electrical S wave of the pulmonary hypertension rat is deepened, which indicates that the right heart function of the animal is damaged. High doses of idenafil tend to suppress the deepening of the S-wave, indicating that it improves cardiac function to some extent.

As shown in figure 2, the systolic pressure of the right ventricle of the rats with pulmonary hypertension of each group is detected, the systolic pressure of the right ventricle of the model animal is obviously increased compared with that of the blank group, and the systolic pressure of the right ventricle of the model animal is reduced by the positive drug bosentan and the high dose of Aidenafil. Compared with normal animals, the Fulton index, namely the right heart hypertrophy index RV/(L + S) (shown as B in figure 2), the right heart index RV/BW (shown as C in figure 2) and the ventricular index (shown as D in figure 2) of the model group are all obviously increased, and meanwhile, the right heart hypertrophy degree of the pulmonary hypertension rat can be obviously reduced by all the dose groups of the aildenafil, and the better dose dependence is presented, which indicates that the citric acid aildenafil has stronger inhibiting effect on the right heart injury of the pulmonary hypertension rat.

Example 4

In the embodiment, from the perspective of preventing pulmonary hypertension, the influence of the citric acid alidenafil on the systemic circulation of the pulmonary hypertension modeled rat is studied.

At the end of the experiment, a left carotid artery blood vessel is cannulated, a median incision is made in the neck of a rat, muscle nerves are separated, blood vessels are stripped, 3 wires are buried, the distal end of the blood vessels is tied by wires, the blood flow is temporarily blocked by wires at the proximal end of the blood vessels, a V-shaped incision is carefully made, a polyethylene plastic tube (PE50) which is filled with heparin sodium solution (100U/mL) in advance is inserted, the other end of the polyethylene plastic tube is connected with a BL420 biological function experimental instrument, the tubes and the blood vessels are fixed by wires, the wires at the proximal end of the blood vessels are loosened, the blood flow is recovered, and the pressure of the main artery including systolic pressure (SBP), diastolic pressure (DBP) and Heart Rate (HR) is recorded after the blood pressure is stabilized.

The systemic circulation of the rat with pulmonary hypertension is monitored. Firstly, observing whether the medicine influences the systemic circulation pressure and the heart rate; secondly, when the pulmonary hypertension develops to a certain degree, the cardiac function is damaged, so that the systemic circulation pressure changes, and therefore, the systemic circulation pressure and the heart rate can reflect the body state of the animal, and the severity of the pulmonary hypertension is indirectly indicated.

The circulation of each group of animals was examined using left carotid vascular intubation, and the results are shown in fig. 3, with significant reductions in systolic and diastolic blood pressure (# P <0.05, # P <0.01) for the model group, indicating that a more severe impairment of the animal's cardiac function has occurred, which also laterally explains the lower than expected RVSP for the model animals. In addition, although the systemic circulation pressure and the heart rate of the animals in each administration group are not obviously different from those in the model group, the systemic circulation pressure and the heart rate are close to those in the normal control group, and the fact that the administration of the medicine can improve the heart function of the animals is proved.

Example 5

In this example, the effect of citric acid alidenafil on the index of each organ of a pulmonary hypertension model rat is studied from the viewpoint of preventing pulmonary hypertension.

The index of each organ extracted in the organ detection step in example 1 was measured. After the administration for 21 days, the organ index of the model group and each administration group animal was changed. The results show that as shown in fig. 4, both the heart index and the lung index of the model animal are significantly increased (## P <0.001) during the development of pulmonary hypertension, which indicates that the heart and lung of the rat are damaged. The administration of positive drugs and low, medium and high doses of the elderfil can obviously reduce the cardiac index, which indicates that the elderfil has protective effect on the heart of animals; while low doses of aildenafil can significantly reduce pulmonary edema, suggesting that the protective effect of low doses of aildenafil on the lungs may be more advantageous.

Example 6

In the embodiment, the influence of the citric acid Aidinafei on the NT-proBNP in the serum and the lung tissue of the pulmonary hypertension modeling rat is researched from the aspect of preventing the pulmonary hypertension.

Detecting the content of NT-proBNP in serum and lung tissues by using an ELISA method, preparing various reagents according to the kit specification, balancing for 30min at room temperature, respectively arranging blank holes, standard sample holes and sample holes to be detected, adding 100 mu L of standard substance or sample to be detected into each hole, shaking uniformly, covering a plate for pasting, incubating for 2h at 37 ℃, removing liquid, and spin-drying; adding 100 μ L of biotin labeled antibody working solution into each well, covering with a plate, incubating at 37 deg.C for 1h, discarding the liquid in the well, spin-drying, adding 200 μ L of detergent into each well, washing for 3 times for 2 min/time; adding 100 mu L of a working solution of horseradish peroxidase marker avidin into each hole, covering a plate for sticking, incubating for 1h at 37 ℃, discarding liquid in the hole, spin-drying, and washing the plate for 5 times according to the operation mode; adding 90 μ L of substrate reaction solution into each well, developing at 37 deg.C in dark for 15-30min, adding 50 μ L of reaction terminating solution, and terminating the reaction. Within 5min, detecting the absorbance at 450nm of the microplate reader.

The level of NT-proBNP in serum and lung tissue may be an important indicator of heart failure in a patient. The serum sample test result is shown in fig. 5, the content of NT-proBNP in the serum of the rat in the model group is obviously increased, and the content of NT-proBNP in the serum of each administration group is obviously reduced, which indicates that the ednafil has obvious inhibition effect on the damage of the heart. NT-proBNP also tends to increase in lung tissue of rats in the model group, but the amount of NT-proBNP in the tissue does not vary significantly compared with serum.

Example 7

In the embodiment, from the aspect of preventing pulmonary hypertension, the influence of the citric acid aildenafil on the pulmonary vascular structure of the pulmonary hypertension model rat is researched.

The pulmonary artery vessels with different diameters extracted in the organ detection step in example 1 are taken, and a masson stain is applied, so that the shapes of pulmonary alveoli and blood vessels can be observed, the thicknesses of the blood vessel walls in three diameter ranges in the lung tissue of the PAH rat are counted, and the remodeling trends of the blood vessels with different diameters are approximately the same, as shown in FIG. 6.

The rat pulmonary alveolar morphological structure of the PAH model group is damaged, and the vascular walls in three diameter ranges are thicker than the pulmonary vascular walls with the same diameter of the rat of the control group. Compared with the PAH model group, the positive drug bosentan group can obviously reduce the thickness of the blood vessel wall of the lung tissue with the blood vessel diameter of more than 100 mu m. Moreover, bosentan also tends to reduce the thickness of the vessel wall of the lung tissue of PAH rats with a vessel diameter of less than 100 μm. The positive drug sildenafil has a tendency to lower the blood vessel wall in three diameter ranges of the lung of PAH model animals.

The biological citric acid aildenafil (15mg/kg, 30mg/kg and 60mg/kg) with three concentrations can obviously reduce the vessel wall thickness of the vessel wall thicknesses with three diameter ranges in the lung tissue of the PAH rat so as to relieve the disease state. Of these, the medium dose group (30mg/kg) had the best effect on pulmonary vessels with a vessel diameter of 50 to 100 μm, while the high dose group had the strongest inhibitory effect on the thickening of pulmonary vessel walls in the remaining diameter range.

Example 8

In this example, from the viewpoint of treating pulmonary hypertension, a rat pulmonary hypertension model is established, and a treatment dosing regimen is performed.

In the examples, 70 SPF-grade SD rats were selected, weighed 200 to 220g, and randomly divided into 7 groups of 10 animals each, and the 7 groups were set as a normal control group, a pulmonary hypertension model group, a positive drug sildenafil group (30mg/kg, oral administration, once a day), a positive drug bosentan group (100mg/kg, oral administration, once a day), a citric acid aildenafil low dose group (15mg/kg, oral administration, once a day), a citric acid aildenafil medium dose group (30mg/kg, oral administration, once a day), and a citric acid aildenafil high dose group (60mg/kg, oral administration, once a day). When the rat pulmonary hypertension model is established in other groups, and when the PAH disease of animals is stably formed on 14 th day after the model is built, day14 begins continuous administration after the model animals are regrouped until the death rate of the model group reaches 70%, the administration frequency is once a day, the weight of the animals is monitored daily and the death condition is recorded.

After the last administration when the mortality rate of the model group reached 70%, the animals to be tested were fasted overnight, weighed the next day and recorded. The experimental end-point detection indexes are as follows:

(1) and taking out the lung, performing perfusion cleaning by using normal saline, sucking water by using filter paper, taking 0.2g of frozen right lung tissue, adding 1mL of PBS (phosphate buffer solution), grinding and homogenizing by using an automatic grinder, centrifuging at 4 ℃ for 10min at 1500g, and performing treatment operation according to a specification for cGMP detection.

(2) Cutting the whole ventricle along the lower edge of the auricle, then cutting the right ventricle along the edge of the ventricular septum, weighing, making a ratio of the right ventricle (R) to the sum of the left ventricle and the ventricular septum (L + S), calculating [ R/(L + S) ], making a ratio R/BW of the right ventricle (R) to the weight, calculating the index of the right ventricle, and calculating the hypertrophy degree of the right ventricle.

Example 9

In this example, the effect of Aidenafil citrate on the body weight of rats modeled after pulmonary hypertension was studied from the viewpoint of treating pulmonary hypertension.

The animals were regrouped 14 days after molding and the administration was started while the body weight was recorded. As shown in FIG. 7, the difference in body weight between the normal group and the model group was significant after 14 days of molding, and the body weight of the molded animals was significantly low. At the end of the experiment, the citric acid alidenafil with medium dosage has a certain trend of improving the weight of the animals.

Example 10

In the embodiment, the influence of the Aidenafil citrate on the survival rate of the pulmonary hypertension molding rat is researched from the aspect of treating the pulmonary hypertension.

As shown in table 2, only 40% of rats in the model group survived after 14 days of single MCT molding and the survival rates of model rats after administration of bosentan and sildenafil were 40% and 60%, respectively, but bosentan did not increase the survival rate of animals but delayed the onset of death to some extent. Sildenafil and the sildenafil citrate of each dose group have been shown to have the effect of improving the survival rate of animals. The survival rate of the pulmonary hypertension model rat can be remarkably improved to 70% by using the high-dose citric acid alidenafil, which is far higher than that of a positive medicine.

Table 2 therapeutic effect rat survival at experimental endpoint

Example 11

In the embodiment, from the aspect of treating pulmonary hypertension, the influence of the citric acid aildenafil on the reconstruction of the right ventricle of a pulmonary hypertension model rat is researched.

And at the end of the experiment, weighing the heart weight of the rat, comparing the heart weight with the corresponding body weight to obtain a heart index, a ventricle index and a right ventricle index, and calculating the Fullton index. As shown in fig. 8, the right heart of the model animals was significantly hypertrophic, and the high dose of citric acid aildenafil showed some tendency of improvement in the cardiac index (a in fig. 8), the ventricular index (B in fig. 8), and the right ventricular index (C in fig. 8).

Example 12

In this example, the effect of Aidenafil citrate on cGMP in pulmonary hypertension modeled rat pulmonary tissue was studied from the viewpoint of treating pulmonary hypertension.

As shown in figure 9, the content of cGMP in the lung tissue of rats in the model group is slightly reduced, and the content of cGMP in the lung tissue of animals in each dose group of the citric acid alidenafil is increased to a certain extent, which shows that the citric acid alidenafil can regulate the level of cGMP in a PDE5/cGMP/PKG pathway by inhibiting the activity of PDE5 so as to realize the treatment of pulmonary hypertension.

Examples 1 to 12 of comprehensive analysis

According to the contents of the embodiments 1 to 7, it can be known that no animal dies when the tested drug citric acid aildenafil is given at different doses, and the mortality rate of the model group animals is 20%, which indicates that the citric acid aildenafil really plays a role in delaying the occurrence and development of PAH, and improves the survival time and survival rate of the model animals.

The results of the detection of the electrocardio-right heart systolic pressure show that the citric acid alidenafil has no obvious effect of reducing the right ventricular systolic pressure (namely pulmonary artery pressure), but has obvious effect of inhibiting the reconstruction phenomenon of the right ventricle, which indicates that the citric acid alidenafil is more important in protecting the heart in the aspect of treating the pulmonary artery hypertension.

In the aspect of organ indexes, the citric acid alidenafil can obviously reduce the conditions of cardiac hypertrophy and pulmonary edema and indirectly reflect the cardiopulmonary protection effect.

In the aspect of pulmonary artery vascular remodeling, pathological tissue section observation shows that the rat pulmonary artery vascular remodeling condition of the PAH model group is serious, pulmonary interstitial edema and pulmonary alveolar structure damage. Statistics on the thicknesses of blood vessel walls in three diameter ranges (<50 μm,50-100 μm and >100 μm) shows that the pulmonary artery blood vessel walls of the rat with pulmonary hypertension are thickened, and the citric acid Aidinafei can obviously inhibit the pulmonary vessel reconstruction phenomenon and has better curative effect compared with positive medicines.

The disease marker NT-proBNP is one of the current auxiliary indicators for PAH diagnosis, and the experimental results show that the content of NT-proBNP in the serum of a model animal is obviously increased, and each dosage of the citric acid alidenafil has an obvious reduction effect on the increase of the disease marker, thereby further proving the inhibition effect of the medicament on right heart injury.

Therefore, the citric acid alidenafil can obviously inhibit the pulmonary artery vascular remodeling phenomenon, has a prominent inhibiting effect on structural and functional damage of the right ventricle, has a protective effect on the remodeling inhibitor of the heart and blood vessels, is an important reason for delaying the occurrence and development of diseases and improving the survival rate of animals, and thus has a good pulmonary hypertension preventing effect.

Continuing to analyze the contents of examples 8-12, when the mortality rate of the model group reaches 70% (i.e. 16/14 days after administration), the experiment is terminated, and the positive drug sildenafil shows a better effect of improving the survival rate of animals. The survival rate of the Aidenafil citrate is obviously improved by each dosage of Aidenafil citrate, and the effect of the high dosage is more prominent than that of the positive drug sildenafil. In addition, the citric acid alidenafil with medium dosage has certain effect of improving the body weight of the rat, and the state of the rat is possibly improved to a certain degree.

The high-dose sildenafil citrate has certain improving effect on cardiac index, ventricular index and right ventricular index, and the cGMP content in the lung tissues of animals with various doses of the sildenafil citrate is increased to a certain extent, so that the effect of adjusting the cGMP level in a PDE5/cGMP/PKG pathway to treat pulmonary hypertension by inhibiting the activity of PDE5 in the sildenafil citrate is reflected.

Although the present invention has been described with reference to the preferred embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the present invention.

Claims

1. Use of Aidinafei citrate for the preparation of a medicament for the prevention and/or treatment of pulmonary hypertension.

2. The use according to claim 1, wherein the Aidinafil citrate protects the heart in preventing pulmonary hypertension by inhibiting right ventricular remodeling, reducing right ventricular hypertrophy, and reducing cardiac index.

3. The use of claim 1, wherein the Aidinafil citrate prevents pulmonary hypertension by inhibiting pulmonary edema and reducing pulmonary index.

4. The use according to claim 1, wherein the Aidenafil citrate is used for preventing pulmonary hypertension by inhibiting the thickening of the wall of pulmonary artery blood vessels with different diameters.

5. The use according to claim 2, wherein the citric acid alidenafil inhibits cardiac damage due to right ventricular remodeling by reducing the level of NT-proBNP in lung tissue and serum.

6. The use of claim 1, wherein the therapeutic effect of Aidenafil citrate on pulmonary hypertension comprises increasing survival and increasing weight in pulmonary hypertension model animals.

7. The use according to claim 6, wherein the Aidenafil citrate effects treatment of pulmonary hypertension by inhibiting the activity of PDE5 to modulate the level of cGMP in the PDE5/cGMP/PKG pathway.

8. The use according to any one of claims 1 to 7, characterized in that the single-use dose of the citric acid Aidinafil for preventing and treating pulmonary hypertension for adults is 1 to 100mg/kg, preferably 2.5 to 10 mg/kg.

9. The use according to claim 8, wherein the medicament for preventing and/or treating pulmonary hypertension comprises a preparation of the Aidinafil citrate compound and a pharmaceutically acceptable carrier or adjuvant, and the preparation is selected from the group consisting of: solutions, suspensions, emulsions, pills, capsules, powders, controlled release, sustained release formulations and microsomal delivery systems.

10. Use according to claim 9, wherein the medicament for the prevention and/or treatment of pulmonary hypertension is in an oral dosage form.