Composition for treating pulmonary arterial hypertension and application thereof

Abstract

The invention belongs to the technical field of pulmonary arterial hypertension treatment medicines, and particularly relates to a composition for treating pulmonary arterial hypertension and application thereof. The composition comprises the following raw materials: pachymic acid B, cinnamic acid, atractylenolide I and glycyrrhizic acid. Experiments prove that for pulmonary arterial hypertension rats, the composition can obviously reverse decrease of the PAT / PET ratio, obviously relieve increase of right ventricular systolic pressure and average pulmonary arterial pressure, obviously relieve right ventricular hypertrophy index and obviously relieve PA myelization and pulmonary fibrosis of the pulmonary arterial hypertension rats. Therefore, the composition can be used for preparing medicines for treating pulmonary arterial hypertension.

Description

Technical Field

[0001] The present invention belongs to the technical field of drugs for treating pulmonary hypertension, and in particular relates to a composition for treating pulmonary hypertension and application thereof. Background Art

[0002] Pulmonary hypertension is a malignant disease characterized by persistently elevated pulmonary artery pressure, which can eventually lead to right heart failure or even death. At present, conventional treatments for pulmonary hypertension mainly include drug therapy and surgical treatment. Surgical treatment has defects such as narrow indications, high risk, and low survival rate. For example, chronic thromboembolic pulmonary hypertension (CTEPH) can undergo pulmonary artery thromboendarterectomy, but the thrombus must be completely organized and located proximally; the 5-year survival rate after lung transplantation is only 45%-50%, and there is a shortage of donors. Even if the operation is successful, long-term use of immunosuppressants (such as cyclosporine) and targeted drugs is still required to maintain hemodynamic stability. In drug treatment, Western medicine mainly uses targeted drugs, including endothelin receptor antagonists, phosphodiesterase-5 inhibitors, prostacyclins, diuretics, anticoagulants, etc. Endothelin receptor antagonists and phosphodiesterase-5 inhibitors mainly relieve symptoms by dilating pulmonary blood vessels or inhibiting vasoconstriction, but they cannot reverse the pulmonary vascular remodeling (such as intimal thickening, luminal stenosis) or endothelial cell damage that has occurred. Recent studies have shown that some Chinese herbal compound prescriptions and monomer components can relieve pulmonary hypertension through multi-target effects. However, Chinese medicine is also difficult to reverse endothelial cell damage or severe vascular remodeling that has already formed, and some patients need to rely on Western medicine to maintain basic hemodynamic stability. Summary of the invention

[0003] In view of the above technical problems, the present invention provides a composition for treating pulmonary hypertension and its application. The composition can significantly improve the right ventricular function of rats with pulmonary hypertension, reduce pulmonary circulation pressure, and effectively improve pulmonary vascular remodeling.

[0004] In order to achieve the above-mentioned object of the invention, the present invention adopts the following technical solutions: A first aspect of the present invention provides a composition for treating pulmonary arterial hypertension, wherein the raw materials include 2 to 3 parts of pachymic acid B, 10 to 15 parts of cinnamic acid, 10 to 15 parts of atractylodes lactone I and 16 to 24 parts of glycyrrhizic acid in proportion by mass.

[0005] Experiments have shown that the composition provided by the present invention has the following effects on pulmonary hypertension: (1) It can significantly reverse the decrease in the PAT / PET ratio caused by pulmonary hypertension. A decrease in the PAT / PET ratio usually indicates impaired right ventricular function, and an increase in the ratio indicates improved right ventricular function; (2) It can significantly relieve the elevation of right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) caused by pulmonary hypertension. RVSP and mPAP are the gold standards for judging pulmonary hypertension. The latter also reflects the overall pressure level of the pulmonary circulation. The decrease of both indicates that pulmonary hypertension has been effectively improved; (3) It can significantly reduce the right ventricular hypertrophy index (RV / LV+S). The decrease of this ratio indicates the alleviation of right ventricular hypertrophy and the improvement of right heart function; (4) It can significantly reduce PA muscularization and pulmonary fibrosis in rats with pulmonary hypertension. The decrease in the areas of vascular muscularization and fibrosis indicates the improvement of pulmonary vascular remodeling.

[0006] The above effects prove that the composition provided by the present invention has the potential to treat pulmonary hypertension.

[0007] In addition, the composition will not produce obvious side effects on the body and has good medication safety.

[0008] Preferably, the raw materials of the composition are proportioned by mass parts and include 2.25-2.75 parts of pachymic acid B, 11.25-13.75 parts of cinnamic acid, 11.25-13.75 parts of atractylenolide I, and 18-22 parts of glycyrrhizic acid.

[0009] Preferably, the raw materials of the composition are proportioned by mass parts and include 2.5 parts of pachymic acid B, 12.5 parts of cinnamic acid, 12.5 parts of atractylenolide I, and 20 parts of glycyrrhizic acid.

[0010] The second aspect of the present invention also provides the application of the above composition in the preparation of a drug for treating pulmonary hypertension.

[0011] Preferably, the drug is an oral preparation, such as tablets, granules, capsules, etc.

[0012] Preferably, the composition of the drug further includes excipients.

[0013] The third aspect of the present invention also provides the application of the above composition in screening compounds in vivo or in vitro that have a reduced PAT / PET ratio, elevated right ventricular systolic pressure, elevated mean pulmonary artery pressure, increased right ventricular hypertrophy index, and cause or exacerbate PA muscularization and pulmonary fibrosis. Compounds with the above effects may be impurities, intermediates or metabolites in drugs or foods. The above effects make them likely to directly trigger pulmonary hypertension. Even if they do not directly cause pulmonary hypertension, they will also cause a certain degree of damage to the heart and lungs. By using the composition of the present invention as a negative control for in vivo or in vitro screening, it will help to discover compounds that may trigger pulmonary hypertension or may damage the heart and lungs through the above effects, so as to purposefully remove such compounds from the final product or reduce their content, thereby improving the product safety.

[0014] The beneficial effects of the present invention are as follows: The present invention firstly discovers that the composition composed of pachymic acid B, cinnamic acid, atractylenolide I and glycyrrhizic acid can significantly reverse the decrease of the PAT / PET ratio, significantly relieve the increase of right ventricular systolic pressure and mean pulmonary artery pressure, significantly reduce the right ventricular hypertrophy index, and can significantly alleviate the PA muscleization and pulmonary fibrosis of rats with pulmonary hypertension. Moreover, the above effects are significantly better than those of traditional Chinese medicine extracts with therapeutic effects on pulmonary hypertension. Therefore, this composition has the potential to treat pulmonary hypertension and can be used to prepare drugs for treating pulmonary hypertension. In addition, the above effects of this composition enable it to be used for screening in vivo or in vitro compounds with a decreased PAT / PET ratio, an increased right ventricular systolic pressure, an increased mean pulmonary artery pressure, an increased right ventricular hypertrophy index, causing or aggravating PA muscleization and pulmonary fibrosis. By purposefully removing such compounds from the final product or reducing their content, the product safety can be improved. Description of the Drawings

[0015] Figure 1 For the RVSP, mPAP ( n =8) and RV / LV+S ( n =6) of rats in the Control group, MCT group, L-LGZG group, H-LGZG group and sildenafil group in Example 9 of the present invention; *, P<0.05; **, P<0.01; Figure 2 For the RVSP, mPAP ( n =8) and RV / LV+S ( n =6) of rats in the Control group, MCT group, L-PCAG group, H-PCAG group, H-LGZG group in Example 9 of the present invention; *, P<0.05; **, P<0.01; Figure 3 For the PAT / PET echocardiogram measurement results of rats in the Control group, MCT group, L-PCAG group, H-PCAG group, H-LGZG group in Example 9 of the present invention ( n =8); *, P<0.05; **, P<0.01; Figure 4 For the lung tissue analysis results of rats in the Control group, MCT group, L-PCAG group, H-PCAG group, H-LGZG group in Example 9 of the present invention; *, P<0.05; **, P<0.01; Figure 5 For the body weight measurement results of rats in the Control group, MCT group, L-PCAG group, H-PCAG group, H-LGZG group in Example 9 of the present invention; **, P<0.01; Figure 6This is the H&E staining results of the liver, spleen, and kidneys of rats in the Control group, MCT group, L-PCAG group, H-PCAG group, and H-LGZG group in Example 9 of the present invention. Detailed implementation manners

[0016] In order to make the objectives, technical solutions, and advantages of the present invention clearer and more understandable, the following further elaborates on the present invention in combination with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and not to limit the present invention. All other implementation manners obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts fall within the protection scope of the present invention.

[0017] In the following examples, Poria cocos, Cinnamon Twig, Atractylodes macrocephala, and Honey-Fried Licorice Root used all comply with the relevant regulations under the entries of each medicinal material in Part I of the Chinese Pharmacopoeia (2020 Edition).

[0018] In the following examples, raw materials, reagents, etc. used, unless otherwise specified, are obtained from commercial channels.

[0019] Example 1 The embodiment of the present invention provides a composition for treating pulmonary hypertension, and its raw materials are: 2.5 parts of pachymic acid B, 12.5 parts of cinnamic acid, 12.5 parts of atractylenolide I, and 20 parts of glycyrrhizic acid.

[0020] Example 2 The embodiment of the present invention provides a composition for treating pulmonary hypertension, and its raw materials are: 2.25 parts of pachymic acid B, 13.75 parts of cinnamic acid, 11.25 parts of atractylenolide I, and 22 parts of glycyrrhizic acid.

[0021] Example 3 The embodiment of the present invention provides a composition for treating pulmonary hypertension, and its raw materials are: 2.75 parts of pachymic acid B, 11.25 parts of cinnamic acid, 13.75 parts of atractylenolide I, and 18 parts of glycyrrhizic acid.

[0022] Example 4 The embodiment of the present invention provides a composition for treating pulmonary hypertension, and its raw materials are: 2 parts of pachymic acid B, 15 parts of cinnamic acid, 10 parts of atractylenolide I, and 24 parts of glycyrrhizic acid Example 5 The embodiment of the present invention provides a composition for treating pulmonary hypertension, and its raw materials are: 3 parts of pachymic acid B, 10 parts of cinnamic acid, 15 parts of atractylenolide I, and 16 parts of glycyrrhizic acid.

[0023] Example 6 An embodiment of the present invention provides a granule for treating pulmonary hypertension, and its preparation method is as follows: Mix the composition of any one of Embodiments 1 to 5 with pregelatinized starch, prepare soft material with 5% PVP aqueous solution, granulate through 18 - 20 meshes, screen through 20 - 22 meshes after drying at 45 - 55 °C, add 0.5% magnesium stearate, mix and then subpackage to obtain the granule.

[0024] Pregelatinized starch can be replaced by other pharmaceutically acceptable granule excipients. Flavoring agents and other excipients can also be added to the granule according to actual needs.

[0025] Example 7 An embodiment of the present invention provides a tablet for treating pulmonary hypertension, and its preparation method is as follows: Mix the composition of any one of Embodiments 1 to 5 with pregelatinized starch and microcrystalline cellulose, perform one - step granulation with 3% hydroxypropyl methylcellulose aqueous solution as the binder, add 3% sodium carboxymethylcellulose, 1% magnesium stearate and 0.5% silicon dioxide, and then press into tablets to obtain the tablets.

[0026] Pregelatinized starch and microcrystalline cellulose can be replaced by other pharmaceutically acceptable fillers, hydroxypropyl methylcellulose can be replaced by other pharmaceutically acceptable binders, sodium carboxymethylcellulose can be replaced by other pharmaceutically acceptable disintegrants, and magnesium stearate and silicon dioxide can be replaced by other pharmaceutically acceptable lubricants and glidants. The tablets can also be film - coated according to clinical needs.

[0027] Example 8 An embodiment of the present invention provides a hard capsule for treating pulmonary hypertension, and its preparation method is as follows: Mix the composition of any one of Embodiments 1 to 5 with pregelatinized starch and microcrystalline cellulose, perform one - step granulation with 3% hydroxypropyl methylcellulose aqueous solution as the binder, screen through 22 - 24 meshes, add 1% magnesium stearate, and load into hard capsule shells to obtain the hard capsules.

[0028] Pregelatinized starch and microcrystalline cellulose can be replaced by other pharmaceutically acceptable fillers, and magnesium stearate can be replaced by other pharmaceutically acceptable glidants.

[0029] Example 9 An embodiment of the present invention provides the effects of the composition of Example 1 on the right ventricular function, pulmonary circulation pressure and lung tissue of rats with pulmonary hypertension.

[0030] 1. Construction, grouping and administration method of the rat model of pulmonary hypertension (PH) 1.1 After adapting 8-week-old male SD rats to the standard feeding method for one week, the experiment was started. The rats were divided into a treatment group and a control group (Control group). Pulmonary hypertension (PH) rat models were induced on the first day of the 21-day experimental procedure by single intraperitoneal injection of monocrotaline (MCT, 60 mg / kg) or a control solvent (a mixture of anhydrous ethanol and normal saline with a volume ratio of 2:8). The rats in the treatment group were divided into a model group (MCT group), a low-dose traditional Chinese medicine extract group (L-LGZG group), a high-dose traditional Chinese medicine extract group (H-LGZG group), and a sildenafil group, with 8 rats in each group. From the 7th day to the 21st day of the experimental procedure, intragastric administration was given daily. The administration dose of the L-LGZG group was 2.97 g / kg / day (calculated as crude drug), the administration dose of the H-LGZG group was 5.94 g / kg / day (calculated as crude drug), and the administration dose of the sildenafil group was 30 mg / kg / day. The rats in the control group and the model group were given an equal amount of normal saline daily from the 7th day to the 21st day. During the experiment, all groups of mice were given ordinary feed (Beijing Speywood Biotechnology Co., Ltd., Co60 irradiated and sterilized maintenance rat feed, SPF-F02-002) and drinking water for free diet.

[0031] Among them, the preparation method of the traditional Chinese medicine extract was as follows: 12 g of Poria cocos, 9 g of Cinnamomum cassia (peeled), 6 g each of Atractylodes macrocephala and roasted Glycyrrhiza uralensis. It was reflux-extracted twice with 6 times the amount of distilled water. The first extraction was for 2.5 hours, and the second extraction was for 1.5 hours. After filtration, the filtrates were combined, vacuum-concentrated and then spray-dried to obtain the extract.

[0032] 1.2 After adapting 8-week-old male SD rats to the standard feeding method for one week, the experiment was started. The rats were divided into a treatment group and a control group (Control group). Pulmonary hypertension rat models were induced on the first day of the 21-day experimental procedure by single intraperitoneal injection of monocrotaline (MCT, 60 mg / kg) or a control solvent (a mixture of anhydrous ethanol and normal saline with a volume ratio of 2:8). The rats in the treatment group were divided into a model group (MCT group), a low-dose composition group (L-PCAG group), a high-dose composition group (H-PCAG group), and a high-dose traditional Chinese medicine extract group (H-LGZG group), with 8 rats in each group. From the 7th day to the 21st day of the experimental procedure, intragastric administration was given daily. The administration dose of the L-PCAG group was 47.5 mg / kg / day, the administration dose of the H-PCAG group was 95 mg / kg / day, and the administration dose of the H-LGZG group was 5.94 g / kg / day (calculated as crude drug). The rats in the control group and the model group were given an equal amount of normal saline daily from the 7th day to the 21st day. During the experiment, all groups of mice were given ordinary feed (Beijing Speywood Biotechnology Co., Ltd., Co60 irradiated and sterilized maintenance rat feed, SPF-F02-002) and drinking water for free diet. Among them, the preparation method of the traditional Chinese medicine extract was the same as that in "1.1".

[0033] 2. Detection Indicators 2.1 Echocardiogram Detection On the 21st day of the experimental procedure, the hemodynamic changes of the rats in each group in "1.2" were evaluated using a Vevo 2100 ultrasound imaging system (FUJIFILM Visual Sonics, Toronto, Canada). The rats were anesthetized by continuously inhaling isoflurane (concentration 1.5% - 3.0%) through a nasal mask, and the pulmonary artery acceleration time (PAT) and pulmonary artery ejection time (PET) were recorded by M-mode ultrasound.

[0034] 2.2 Hemodynamic Measurement, Tissue Collection and Morphological Analysis Hemodynamic Parameter Recording On the 21st day of the experimental procedure, before sacrificing the rats in each group after echocardiogram detection, the right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) were recorded and analyzed using a PowerLab physiological data acquisition system (AD Instruments, Australia).

[0035] Tissue Processing and Pathological Evaluation After completing the hemodynamic detection, the rats were sacrificed, and the heart, lungs, liver, spleen, and kidneys were collected. Six samples were randomly selected from the rat hearts in each group, and the degree of right ventricular hypertrophy was evaluated by calculating the ratio of the right ventricle weight to the sum of the left ventricle and interventricular septum weights (RV / LV+S).

[0036] Some lung tissues of the rats in each group in "1.2" were fixed with 4% paraformaldehyde, embedded in paraffin, and sectioned (thickness 5 μm). Van Gieson (VG) staining (evaluating elastic fibers) and Masson staining (detecting collagen deposition) were performed according to the standard procedure.

[0037] The liver, spleen, and kidney tissues of the rats in each group in "1.2" were fixed with 4% paraformaldehyde, embedded in paraffin, and sectioned (thickness 5 μm). H&E staining was performed according to the standard procedure.

[0038] 2. Results 2.1 Hemodynamic Results and Right Ventricular Hypertrophy Index As Figure 1 shown: The hemodynamic results of the rats in each group in "1.1" showed that compared with the control group, the RVSP and mPAP of the rats in the MCT group were significantly increased (both P<0.01). The low and high doses of the traditional Chinese medicine extract LGZG and sildenafil significantly decreased the RVSP and mPAP (P<0.01). The RV / LV+S results of the rats were consistent with the hemodynamic results.

[0039] As Figure 2As shown, the hemodynamic results of each group of rats in "1.2" showed that PCAG could significantly alleviate the elevation of RVSP and mPAP caused by MCT (P<0.01). It is worth noting that compared with the H-LGZG group, the improvement of RVSP in the H-PCAG group was more obvious (P<0.01). The evaluation of RV / LV+S showed that PCAG significantly alleviated right ventricular hypertrophy in PH rats (P<0.01). In addition, compared with H-LGZG, the decrease in RV / LV+S caused by H-PCAG treatment was greater (P<0.01).

[0040] Note: Since the hemodynamic results of each group of rats in "1.1" and "1.2" were completed at different times and measured by different instruments respectively, there were batch-to-batch data differences, but it did not affect the within-batch data.

[0041] 2.2 Echocardiogram detection results As Figure 3 shown, PCAG could significantly reverse the decrease in the PAT / PET ratio caused by MCT.

[0042] 2.3 Lung tissue analysis As Figure 4 shown, PCAG could significantly alleviate PA muscleization (Figure A) and pulmonary fibrosis (Figure B) in PH rats.

[0043] 2.4 Body weight and H&E staining results.

[0044] As Figure 5 shown, compared with the control group, the body weight of rats in the MCT group was significantly reduced (P<0.01), and the traditional Chinese medicine extract LGZG and the composition PCAG had no adverse effects on the body weight of pulmonary hypertension rats.

[0045] As Figure 6 shown, the traditional Chinese medicine extract LGZG and the composition PCAG had no adverse effects on the liver, spleen, and kidney tissues of pulmonary hypertension rats.

[0046] From the above results, it can be seen that the composition of Example 1 can significantly improve the right ventricular function of pulmonary hypertension rats, reduce the pulmonary circulation pressure, effectively improve pulmonary vascular remodeling, promote lung tissue rehabilitation, and have no obvious side effects.

[0047] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, or improvements made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. A composition for treating pulmonary hypertension, characterized in that, The raw materials of the composition are proportioned by mass parts and include 2-3 parts of pachymic acid B, 10-15 parts of cinnamic acid, 10-15 parts of atractylenolide I, and 16-24 parts of glycyrrhizic acid.

2. The composition according to claim 1, wherein The raw materials of the composition are proportioned by mass parts and include 2.25-2.75 parts of pachymic acid B, 11.25-13.75 parts of cinnamic acid, 11.25-13.75 parts of atractylenolide I, and 18-22 parts of glycyrrhizic acid.

3. The composition according to claim 2, wherein The raw materials of the composition are proportioned by mass parts and include 2.5 parts of pachymic acid B, 12.5 parts of cinnamic acid, 12.5 parts of atractylenolide I, and 20 parts of glycyrrhizic acid.

4. Use of the composition according to any one of claims 1-3 in the preparation of a drug for treating pulmonary hypertension.

5. The application according to claim 4, wherein The drug is an oral preparation.

6. The application according to claim 4, characterized in that The components of the drug further include a pharmaceutically acceptable excipient.

7. Use of the composition according to any one of claims 1-3 in screening for a compound having at least one of the following effects in vivo or in vitro: Reducing the PAT / PET ratio; Increasing the right ventricular systolic pressure; Increasing the mean pulmonary artery pressure; Increasing the right ventricular hypertrophy index; Causing or exacerbating PA muscularization and pulmonary fibrosis.

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