Application of licochalcone B as Furin enzyme inhibitor

Abstract

The invention relates to the technical field of medicines, and relates to application of licochalcone B as a Furin enzyme inhibitor. In particular to application of licochalcone B in preparation of drugs for preventing, relieving and / or treating diseases related to Furin enzyme activity increase. The application shows that the licochalcone B has a certain inhibition effect on Furin enzyme activity, has a good pharmacodynamic effect on hypoxia-induced pulmonary arterial hypertension and monocrotaline-induced pulmonary arterial hypertension, can obviously reduce right ventricular systolic pressure, improve right heart remodeling and pulmonary artery blood flow, inhibit pulmonary artery blood vessel remodeling, and can be used for treating pulmonary arterial hypertension. In addition, the Furinase signal channel in the lung tissue of the pulmonary arterial hypertension disease can be obviously reduced. Licochalcone B has good application and development prospects, and is an ideal compound for treating pulmonary arterial hypertension and complications thereof.

Description

Technical Field

[0001] The present invention relates to the field of medical technology and provides a use of licochalcone B as a furinase inhibitor. Licochalcone B has strong furinase inhibitory activity and can be used to prepare a drug for preventing, alleviating and / or treating pulmonary hypertension. Background Art

[0002] Licochalcone B (CAS number: 58749-23-8) is a licorice flavonoid extracted from licorice. Licochalcone B has a wide range of biological activities, including anti-tumor, anti-inflammatory, and antiviral effects (DOI: 10.15975 / j.cnki.gsyy.2023.10.002). However, there are no reports of its inhibitory effects on furin or its potential for the prevention and treatment of pulmonary hypertension.

[0003] Furin is a major protein convertase in the exocrine pathway, located outside the Golgi apparatus, catalyzing the cleavage of multiple protein precursors to produce mature proteins. Currently, the pharmaceutical uses of furin inhibitors are mainly focused on viral infectious diseases, especially the prevention and treatment of the new coronavirus (PMID: 37481137). Inhibiting furin activity may also have therapeutic effects on cancers such as endometrial cancer (PMID: 38018319). Furin is a key protease for the maturation and activation of TGFβ. Its activation can upregulate the TGFβ / Smad signaling pathway, which is a key signaling pathway mediating the development of pulmonary arterial vascular remodeling. Therefore, reducing furin activity can inhibit the development of pulmonary hypertension (PMID: 17673680).

[0004] Pulmonary hypertension is a rare but very serious disease. It refers to a hemodynamic and pathophysiological state in which the pulmonary artery pressure increases beyond a certain value. The current diagnostic standard is that at rest at sea level, the mean pulmonary artery pressure mPAP is ≥ 25 mmHg detected by right cardiac catheterization (Guidelines for the Diagnosis and Treatment of Pulmonary Hypertension in China (2021 Edition)). The age of the population affected is relatively young, with about 75% of patients concentrated in the 20-40 age group, and about 15% of patients are under 20 years old. At present, there is still no clear conclusion on the cause of pulmonary hypertension. Primary proliferative lesions of pulmonary arterioles leading to vascular occlusion may be the main cause of its onset.

[0005] Pulmonary hypertension and hypertension are two distinct diseases. Pulmonary hypertension primarily involves structural and functional impairment of the pulmonary arteries, while hypertension is a clinical syndrome characterized by elevated systemic arterial blood pressure. The two diseases belong to different systems, with distinct pathogenesis, pathological manifestations, and treatments. Currently available therapeutic agents for the two diseases are completely different. When pulmonary hypertension develops, patients experience symptoms such as dyspnea, palpitations, chest pain, hemoptysis, and syncope. In advanced stages, they may develop hypoxia, right ventricular hypertrophy, and even right ventricular failure, and in severe cases, death may occur. Currently, treatment options and medications for this disease are severely lacking, and the prognosis is dismal. Only 20% of patients can achieve adequate control, and most die within 2-3 years of diagnosis. The current first-line treatment for pulmonary hypertension is the endothelin receptor antagonist bosentan, but its effectiveness is unsatisfactory. The development of new, effective drugs for the treatment of pulmonary hypertension is a crucial and urgent task. Summary of the Invention

[0006] The purpose of the present invention is to provide an application of licorice chalcone B in inhibiting furin enzyme.

[0007] Another object of the present invention is to provide a use of licochalcone B in the preparation of a medicament for treating and / or preventing diseases requiring inhibition of furin enzyme activity.

[0008] Another object of the present invention is to provide a use of licochalcone B in the preparation of a drug for reducing the production of mature TGFβ and activation of the TGFβ / Smad signaling pathway in patients with pulmonary hypertension.

[0009] Another object of the present invention is to provide a use of licochalcone B in the preparation of a drug for inhibiting pulmonary artery vascular remodeling and delaying the occurrence and development of pulmonary hypertension.

[0010] Another object of the present invention is to provide a use of licochalcone B in the preparation of a medicament for alleviating, treating and / or preventing pulmonary hypertension complications and other related diseases.

[0011] To achieve the purpose of the present invention, the present invention provides the following technical solutions:

[0012] The first aspect of the technical solution of the present invention is to provide the use of licorice chalcone B as shown in formula I in the preparation of a drug for inhibiting the activity of Furin enzyme.

[0013]

[0014] The second aspect of the technical solution of the present invention is to provide the use of licochalcone B in the preparation of a drug for preventing, alleviating and / or treating diseases related to the inhibition of furin enzyme activity.

[0015]

[0016] The disease is pulmonary hypertension and its complications.

[0017] The pulmonary hypertension and its complications are selected from hypoxic pulmonary hypertension and arterial pulmonary hypertension.

[0018] The prevention, alleviation and / or treatment of pulmonary hypertension is achieved by reducing the activity or expression of furin enzyme, lowering pulmonary artery pressure, improving pulmonary artery vascular remodeling and inhibiting right heart remodeling through licochalcone B.

[0019] The pulmonary hypertension and its complications are selected from cor pulmonale and heart failure.

[0020] The third aspect of the technical solution of the present invention is to provide a pharmaceutical composition for use in preparing a drug for preventing, alleviating and / or treating diseases requiring inhibition of furin enzyme activity, characterized in that the pharmaceutical composition contains a therapeutically effective dose of a compound represented by general formula (I) and a pharmaceutically acceptable carrier or excipient.

[0021]

[0022] The pharmaceutical composition is selected from the following dosage forms: solution, suspension, freeze-dried powder injection, emulsion, pill, capsule, powder, controlled release, sustained release preparation and microsome delivery system.

[0023] The pharmaceutical composition also includes a pharmaceutical composition prepared by combining with other substances in any proportion. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 .Inhibitory effect of licorice chalcone B on furin enzyme activity.

[0025] Figure 2 Effects of licochalcone B on right ventricular systolic pressure in a mouse model of pulmonary hypertension induced by 2 weeks of simple hypoxia.

[0026] Figure 3 Pharmacological effects of licorice chalcone B on a mouse model of pulmonary hypertension induced by SU5416 and 4 weeks of hypoxia. (A is right ventricular systolic pressure, B is right ventricular index)

[0027] Figure 4 Pharmacological effects of licorice chalcone B on a rat model of monocrotaline-induced pulmonary hypertension. (A is right ventricular systolic pressure, B is right ventricular index, C is pulmonary artery maximum blood flow velocity, D is right ventricular internal diameter, E is pulmonary artery blood flow ultrasound image, and F is ventricular ultrasound image)

[0028] Figure 5 Licochalcone B protects the lungs of a rat model of monocrotaline-induced pulmonary hypertension. (A is a Masson-stained lung tissue section, B is the lung index, and C is a statistical chart of the relative thickness of the pulmonary artery wall.)

[0029] Figure 6 Effects of Licorice Chalcone B on the Furin enzyme signaling pathway in the lung tissue of a rat model of monocrotaline-induced pulmonary hypertension. (A is a representative Western blot band, B is a statistical graph of Furin enzyme protein expression, and C is a statistical graph of the ratio of mature TGFβ1 to TGFβ1 precursor expression.)

[0030] Figure 7 Effects of Licorice Chalcone B on the Furin enzyme signaling pathway in the right ventricular tissue of a rat model of monocrotaline-induced pulmonary hypertension. (A is a representative Western blot band; B is a statistical chart of Furin enzyme protein expression) DETAILED DESCRIPTION

[0031] The following examples further illustrate the inhibitory effect of Licorice Chalcone B on Furin enzyme and its pharmacological effect of alleviating the occurrence and development of pulmonary hypertension.

[0032] The following examples are only used to illustrate the present invention in detail and are not intended to limit the present invention in any way.

[0033] Example 1 Licorice Chalcone B Inhibits Furin Enzyme Activity

[0034] The experiment was conducted in a 384-well black plate. The reaction volume was 50 μl, the final concentration of furin was 0.5 μg / ml, the final concentration of the substrate pERTKR-AMC was 16.2 μM, and the final concentration of licorice chalcone B was 100, 30, 10, 3, and 1 μM. The DMSO content in the reaction system was less than 1 / 1000. In kinetic monitoring mode, fluorescence intensity was measured every 3 minutes for 60 minutes. The difference in fluorescence intensity over 60 minutes was used to calculate the reaction rate for each well, representing the approximate initial velocity of the enzymatic reaction.

[0035] The initial reaction velocity of the sample wells is represented by V, the initial reaction velocity of the negative control wells is represented by Vc, and the initial reaction velocity of the blank control wells is represented by Vb. The inhibition rate of the compound on the enzyme is calculated as follows: Inhibition rate % = (1-(V-Vc) / (Vb-Vc))*100%. The results show that the IC50 of licorice chalcone B is approximately 33.76μM. (Appendix Figure 1 )

[0036] Example 2 Pharmacological Effects of Licochalcone B on Hypoxic Pulmonary Hypertension Mouse Model

[0037] Forty male C57 mice weighing 18-22g were divided into four groups: normal, model, sildenafil, and licochalcone B, with 10 mice in each group. After acclimation, the sildenafil and licochalcone B groups were given prophylactic medication for three days at a dose of 42mg / kg. The normal and model groups were given an equal volume of blank solvent. On the fourth day, all animals except the normal group were placed in a hypoxic chamber maintained at 10% oxygen. The treated groups continued to receive medication daily. Right ventricular systolic pressure was measured 14 days later.

[0038] The results showed that after 2 weeks of hypoxia, the right ventricular systolic pressure of the model group animals increased significantly, and the administration of licorice chalcone B could reduce the right ventricular systolic pressure of the animals. (Appendix Figure 2 )

[0039] Example 3 Pharmacological Effects of Licorice Chalcone B on Hypoxia Combined with SU5416-Induced Pulmonary Hypertension in Mice

[0040] Forty male C57 mice weighing 18-22 g were divided into four groups: normal, model, sildenafil, and licochalcone B, with 10 mice in each group. After acclimation, all animals except the normal group were subcutaneously injected with 20 mg / kg SU5416 and then placed in a hypoxic chamber to maintain an oxygen concentration of 10%. SU5416 was administered subcutaneously once weekly. Two weeks later, the sildenafil and licochalcone B groups were initially treated with 42 mg / kg sildenafil or licochalcone B, while the control group received an equal volume of blank solvent. After two weeks of continuous administration, right ventricular systolic pressure and right cardiac remodeling were measured.

[0041] The results showed that licorice chalcone B could significantly reduce the right ventricular systolic pressure and right heart index of animals. (Appendix Figure 3 )

[0042] Example 4 Pharmacological Effects of Licorice Chalcone B on Monocrotaline-Induced Pulmonary Hypertension Rat Model

[0043] Crotaline is a pyrrolidine alkaloid extracted from lily seeds. It is converted into lily pyrrole by the mixed-function oxidase in the liver of rats. Lily pyrrole damages the pulmonary vascular endothelial cells, thereby causing progressive proliferation of pulmonary artery smooth muscle cells, leading to a progressive increase in pulmonary artery pressure, and creating a rat pulmonary hypertension model.

[0044] Forty SD rats were randomly divided into four groups, each with 10 rats: a normal control group, a model group, a positive drug bosentan group, and a licoricechalcone B-treated group. Except for the normal control group, all other groups received a single subcutaneous injection of 50 mg / kg monocrotaline to establish a rat pulmonary hypertension model. The normal control group received an equal volume of saline. Drug administration began the day after modeling and continued for 21 consecutive days with either bosentan or licoricechalcone B at a dose of 30 mg / kg. The normal and model groups received an equal volume of solvent. At the end of the experiment, echocardiography was used to assess pulmonary artery blood flow and heart function in each group, followed by measurement of right ventricular systolic pressure via the right external jugular vein. After sacrifice, the degree of right ventricular thickening, lung index, morphological changes in lung vascular tissue, and protein expression in lung tissue were assessed to determine the efficacy of licoricechalcone B in rat pulmonary hypertension and its potential use in the preparation of products for the prevention and treatment of pulmonary hypertension and its complications.

[0045] The results showed that licorice chalcone B could significantly reduce the right ventricular systolic pressure in rats with pulmonary hypertension, inhibit right ventricular wall hypertrophy and right ventricular dilatation, and thus inhibit right heart remodeling. Furthermore, licorice chalcone B also showed a significant improvement in pulmonary artery blood flow. (Appendix Figure 4 )

[0046] Pathological sections of lung tissue showed that the model animals had pulmonary edema and collagen deposition, as well as thickening of the pulmonary artery walls and vascular remodeling. The licorice chalcone B-treated group significantly alleviated this phenomenon. Testing of lung indexes also showed that licorice chalcone B could significantly inhibit pulmonary edema. (Appendix Figure 5 )

[0047] In order to explore the mechanism of action of licorice chalcone B based on the furin enzyme signaling pathway, the expression levels of related proteins in lung tissue were detected. The results of Western blotting experiments showed that licorice chalcone B could significantly reduce the expression level of furin enzyme in the lung tissue of model animals, reduce the maturation and activation ratio of TGFβ1, inhibit the TGFβ signaling pathway, and thus inhibit pulmonary artery vascular remodeling. In addition, at the transcriptional level, furin and TGFβ1 also showed the same trend. (Appendix Figure 6 )

[0048] Since licochalcone B can significantly improve right heart remodeling, inhibit right ventricular systolic pressure, inhibit right ventricular wall hypertrophy and right ventricular dilatation, the expression levels of related proteins in right ventricular wall tissue were detected. The results of Western blotting experiments showed that licochalcone B can significantly reduce the expression level of Furin enzyme in the lung tissue of model animals, which is consistent with the results in lung tissue. (Appendix Figure 7 ).

Claims

1. Use of licorice chalcone B as shown in formula I in the preparation of a drug for inhibiting furin enzyme activity, 2. Use of licochalcone B in the preparation of a medicament for preventing, alleviating and / or treating diseases requiring inhibition of furin enzyme activity, 3. The method according to claim 2, wherein the disease is pulmonary hypertension and its complications.

4. The method according to claim 3, wherein: The pulmonary hypertension and its complications are selected from hypoxic pulmonary hypertension and arterial pulmonary hypertension.

5. The method according to claim 4, characterized in that The prevention, alleviation and / or treatment of pulmonary hypertension is achieved by reducing the activity or expression of furin enzyme, lowering pulmonary artery pressure, improving pulmonary artery vascular remodeling and inhibiting right heart remodeling through licochalcone B.

6. The method according to claim 3, wherein: The pulmonary hypertension and its complications are selected from cor pulmonale and heart failure.

7. Use of a pharmaceutical composition in the preparation of a drug for preventing, alleviating and / or treating a disease requiring inhibition of furin enzyme activity, characterized in that: The pharmaceutical composition contains a therapeutically effective dose of the compound represented by general formula (I) and a pharmaceutically acceptable carrier or excipient.

8. Use according to claim 7, characterized in that The pharmaceutical composition is selected from the following dosage forms: solution, suspension, freeze-dried powder injection, emulsion, pill, capsule, powder, controlled release, sustained release preparation and microsome delivery system.

9. The method according to claim 7, wherein: The pharmaceutical composition also includes a pharmaceutical composition prepared by combining the pharmaceutical composition with other substances in any proportion.

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