Application of Nrf3 inhibitor in preparation of medicine for preventing and / or treating hypertension

Abstract

The invention provides application of an Nrf3 inhibitor in preparation of a medicine for preventing and / or treating hypertension, and belongs to the field of medical treatment and medicine. The invention finds that the expression level of Nrf3 in the aortic tissue of a hypertension mouse is obviously increased, and the Nrf3 inhibitor can obviously improve the occurrence of hypertension and vascular dysfunction, and can be used for preparing the medicine for preventing and / or treating hypertension.

Description

Technical Field

[0001] The present invention belongs to the field of medical treatment and medicine, and particularly relates to the use of an Nrf3 inhibitor in the preparation of a drug for preventing and / or treating hypertension. Background Art

[0002] Hypertension refers to a clinical syndrome characterized by increased systemic arterial blood pressure (systolic and / or diastolic blood pressure) (systolic blood pressure ≥ 140 mmHg and / or diastolic blood pressure ≥ 90 mmHg), which may be accompanied by functional or organic damage to organs such as the heart, brain, and kidneys. Hypertension is the most common chronic disease and the main risk factor for cardiovascular and cerebrovascular diseases. Existing hypertension drugs can effectively control the blood pressure of most patients, but clinically there are still some patients who are resistant to existing treatments. The pathogenesis of hypertension is complex, and the existing mechanisms include neural mechanisms, renal mechanisms, hormonal mechanisms, vascular mechanisms, and insulin resistance. In recent years, more and more evidence has shown that immune inflammatory responses play an important role in the pathogenesis of hypertension. Some of the pathogenic factors of hypertension include high levels of angiotensin II and aldosterone, as well as a high-salt diet, which activate innate and adaptive immunity, activate immune cells such as macrophages, B / T cells, and dendritic cells, and promote the release of inflammatory factors such as interleukin (IL)-17, IL-6, interferon-γ, and autoantibodies, thereby damaging organs such as the kidneys and blood vessels. In addition, inflammatory factors also promote the production of reactive oxygen species by endothelial cells, reduce the level of nitric oxide, and lead to impaired endothelial cell-dependent vasodilation function. The mechanism by which inflammatory responses are involved in hypertension is quite complex. A single anti-inflammatory treatment target sometimes fails to reduce the overall probability of cardiovascular events and also poses the risk of infection. Therefore, there is still an urgent need to discover new pathogenesis of hypertension, find new treatment targets, and achieve precision treatment.

[0003] The transcription factor Nrf3 (Nuclear factor erythroid 2-related factor 3) is abundantly expressed in the cornea, skin, bladder, and placenta. While it is commonly expressed in other tissues, the expression levels are lower. Nrf3 has been implicated in cancer progression, among other factors, but its mechanism of action in hypertension has not yet been reported. Summary of the Invention

[0004] In order to solve the problems existing in the prior art, the object of the present invention is to provide the use of Nrf3 inhibitors in the preparation of drugs for preventing and / or treating hypertension.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides the use of an Nrf3 inhibitor in preparing a medicament for preventing and / or treating hypertension.

[0007] Preferably, the inhibition rate of the Nrf3 inhibitor on the biological activity of Nrf3 is 50% to 100%.

[0008] Preferably, the Nrf3 inhibitor includes an HIV-1 protease inhibitor and / or a peptide vinyl sulfone compound.

[0009] Preferably, the HIV-1 protease inhibitor includes any one or more of ritonavir, nelfinavir and saquinavir.

[0010] Preferably, the peptide vinyl sulfone compound includes WRR139.

[0011] Preferably, the Nrf3 inhibitor alleviates arterial diastolic dysfunction caused by hypertension.

[0012] Preferably, the hypertension is angiotensin II-induced hypertension.

[0013] Preferably, the active ingredient of the drug includes an Nrf3 inhibitor.

[0014] Preferably, the drug further comprises pharmaceutically acceptable excipients.

[0015] Preferably, the dosage form of the drug includes any one or more of oral preparations, intravenous injections and intramuscular injections.

[0016] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0017] The present invention discovered for the first time that the expression level of Nrf3 in the aorta tissue of hypertensive mice was significantly increased, and experimentally confirmed that Nrf3 inhibitors can significantly improve the occurrence of hypertension and vascular dysfunction, providing a basis for the preparation of drugs for the prevention and / or treatment of hypertension. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 : Blood pressure test results of the model group and the control group;

[0019] Figure 2 :Detection results of Nrf3 expression levels in aorta of angiotensin II-induced hypertensive mice and healthy control mice;

[0020] Figure 3 :Experimental results on the prevention of angiotensin II-induced hypertension in mice by injection of Nrf3 inhibitors;

[0021] Figure 4:Experimental results of preventing angiotensin II-induced vasodilation dysfunction in mice by injection of Nrf3 inhibitor. DETAILED DESCRIPTION

[0022] The present invention provides the use of Nrf3 inhibitors in the preparation of medicaments for the prevention and / or treatment of hypertension. "Nrf3 inhibitors" herein refer to any compound capable of partially or completely inhibiting the biological activity of Nrf3, preferably products that inhibit Nrf3 nuclear translocation or inhibit the expression of target genes activated by Nrf3. Preferably, the Nrf3 inhibitors described herein have an inhibition rate of 50% to 100% on the biological activity of Nrf3, and preferably an inhibition rate of at least 60%, 70%, 75%, 80%, 85%, 90%, or 95%. The present invention has found that Nrf3 expression levels are significantly elevated in the aorta tissue of hypertensive mice. Nrf3 inhibitors can significantly improve the development of hypertension and vascular dysfunction. Therefore, Nrf3 inhibitors can be used to prepare medicaments for the prevention and / or treatment of hypertension.

[0023] The Nrf3 inhibitors described herein include HIV-1 protease inhibitors and / or peptide vinyl sulfone compounds. The HIV-1 protease inhibitors preferably include any one or more of ritonavir, nelfinavir, and saquinavir, including but not limited to optional embodiment 1: ritonavir, nelfinavir, or saquinavir are used alone; optional embodiment 2: ritonavir, nelfinavir, and saquinavir are used in combination in pairs; and optional embodiment 3: ritonavir, nelfinavir, and saquinavir are used in combination. The peptide vinyl sulfone compound preferably includes WRR139.

[0024] The Nrf3 inhibitor of the present invention can inhibit the increase in blood pressure induced by angiotensin II and alleviate arterial diastolic dysfunction caused by hypertension.

[0025] The active ingredient of the drug of the present invention includes an Nrf3 inhibitor. The Nrf3 inhibitor can be used as the sole active ingredient, or the Nrf3 inhibitor can be used in combination with other products for alleviating / improving / treating hypertension.

[0026] The drug of the present invention further includes pharmaceutically acceptable excipients, the addition of which is used to ensure the convenient preparation and clinical application of the prepared pharmaceutical preparation. Further preferably, the pharmaceutical excipients of the present invention include but are not limited to at least one or more of diluents, wetting agents, adhesives, lubricants, colorants, and coating agents.

[0027] The dosage form of the drug of the present invention includes any one or more of oral preparations, intravenous injections and intramuscular injections. The drug of the present invention can be optionally administered enterally, preferably an oral preparation; the oral preparation includes but is not limited to tablets, capsules, syrups, etc., and the oral preparation of the present invention is more preferably used in the form of a controlled-release preparation. The drug of the present invention can be optionally formulated into a preparation for parenteral administration, preferably in the form of a sterile solution suitable for intravenous or intramuscular administration, including an intravenous injection or an intramuscular injection. The dosage form of the drug of the present invention is not limited thereto, and other achievable dosage forms are within the scope of protection of the present invention. The drug of the present invention can be prepared according to conventional methods.

[0028] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0029] In the following examples, unless otherwise specified, all methods are conventional.

[0030] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0031] Example 1

[0032] 1. Construction of animal model of hypertension:

[0033] Male SPF C57BL / 6 mice aged 8 to 10 weeks weighing 25 to 30 g were injected subcutaneously with angiotensin II for 2 weeks at a dose of 490 ng / (kg . min), during which blood pressure was measured every 2 days; a normal saline-treated control group was also set up: 8-10-week-old male SPF-grade C57BL / 6 mice weighing 25-30 g were subcutaneously implanted with a pump and stimulated with normal saline for 2 weeks, during which blood pressure was measured every 2 days.

[0034] Blood pressure was measured using the mouse tail method: the mouse was placed in a tube, with the tail exposed and placed in the rubber tube. The mouse was then placed on a 37°C heating blanket for 5 minutes before blood pressure testing began. After the blood pressure of the mouse stabilized, at least 5 consecutive measurements were recorded. The average of all values ​​was used as the blood pressure of the mouse. The blood pressure test results of the model group and the control group are shown below. Figure 1 As shown, the results showed that angiotensin II-induced hypertension model in mice was successfully established.

[0035] 2. Collect the mouse aorta

[0036] After the mice were anesthetized, the aortas of angiotensin II-induced hypertensive mice (n=5) and saline-treated control mice were harvested and immediately cooled in liquid nitrogen and stored at -80°C.

[0037] 3. qPCR detection

[0038] Extract mouse RNA: Add Trizol to mouse aorta tissue and lyse the tissue with magnetic beads. Add chloroform to extract RNA. After precipitation with isopropanol, wash twice with 75% ethanol, discard the supernatant, dry thoroughly, and dissolve the RNA in water.

[0039] RNA was reverse transcribed into cDNA using a reverse transcription kit (purchased from Takara, product number RR047A). TM The mRNA level of the Nrf3 gene was detected by Multicor Real-Time PCR detection system (Bio-Rad) and normalized with the mRNA level of the housekeeping gene GAPDH (Glyceraldehyde-3-phosphate dehydrogenase).

[0040] Test results such as Figure 2 As shown, the results showed that the expression level of Nrf3 in the aorta of angiotensin II-induced hypertensive mice was upregulated compared with that in healthy control mice, indicating that the expression level of Nrf3 in the aorta was increased in the animal model of hypertension.

[0041] Example 2

[0042] Efficacy of Nrf3 inhibitors in angiotensin II-induced mouse model of hypertension:

[0043] 1. Establishment and grouping of angiotensin II-induced hypertension model in mice

[0044] Male SPF C57BL / 6 mice, weighing 25-30 g and aged 8-10 weeks, were administered angiotensin II via a subcutaneous implanted pump for 2 weeks (the day of hypertension modeling was considered day 0, and simultaneous intervention began on day 1). A control group of mice was treated with normal saline. Treatments for each group were as follows:

[0045] (1) Nrf3 inhibitor-nelfinavir experimental group:

[0046] Control group 1: physiological saline + solvent control: 5 mice, raised for 2 weeks; solvent control (physiological saline containing 4% DMSO) was injected intraperitoneally once a day starting from day 1, for a total of 14 injections.

[0047] Control group 2: saline + nelfinavir: 5 mice, raised for 2 weeks; nelfinavir (purchased from Tocris Bioscience, product number 3766, 100 mg / kg, Nrf3 inhibitor dissolved in saline containing 4% DMSO for injection) was injected intraperitoneally once a day starting from day 1, for a total of 14 injections.

[0048] Modeling group 1: Angiotensin II + solvent control: 10 mice, raised for 2 weeks; solvent control (normal saline containing 4% DMSO) was injected intraperitoneally once a day starting from day 1, for a total of 14 injections.

[0049] Modeling group 2: angiotensin II + nelfinavir: 7 mice, raised for 2 weeks; nelfinavir (purchased from Tocris Bioscience, product number 3766, 100 mg / kg, Nrf3 inhibitor dissolved in saline containing 4% DMSO for injection) was injected intraperitoneally once a day starting from day 1, for a total of 14 injections.

[0050] (2) Nrf3 inhibitor-WRR139 experimental group:

[0051] Control group 1: physiological saline + solvent control: 6 mice, raised for 2 weeks; solvent control (physiological saline containing 4% DMSO) was injected intraperitoneally once a day starting from day 1, for a total of 14 injections.

[0052] Control group 2: saline + WRR139: 7 mice, raised for 2 weeks; WRR139 (purchased from MedChemExpress, product number 2138924-36-2, 100 mg / kg, WRR139 dissolved in saline containing 4% DMSO for injection) was injected intraperitoneally once a day starting from day 1, for a total of 14 injections.

[0053] Modeling group 1: Angiotensin II + solvent control: 6 mice, raised for 2 weeks; solvent control (normal saline containing 4% DMSO) was injected intraperitoneally once a day starting from day 1, for a total of 14 injections.

[0054] Modeling group 2: Angiotensin II + WRR139: 7 mice, raised for 2 weeks; WRR139 (purchased from MedChemExpress, product number 2138924-36-2, 100 mg / kg, WRR139 dissolved in saline containing 4% DMSO for injection) was injected intraperitoneally once a day starting from day 1, for a total of 14 injections.

[0055] 2. Determination of blood pressure in mice

[0056] Step 1: Measure blood pressure every two days in each group of mice during treatment. Measure blood pressure using the tail method: Place the mouse in a tube, then place the tail inside the rubber tube. Place the mouse on a 37°C heating blanket for 5 minutes before measuring blood pressure. Once blood pressure stabilizes, record at least five consecutive measurements. The average of all values ​​is used as the mouse's blood pressure.

[0057] Figure 3 Results of an experiment in which mice were injected with Nrf3 inhibitors to prevent angiotensin II-induced hypertension. Figure A shows systolic blood pressure in mice injected with nelfinavir or a vehicle control, given saline or angiotensin II via a subcutaneous implanted pump; B shows diastolic blood pressure in mice injected with nelfinavir or a vehicle control, given saline or angiotensin II via a subcutaneous implanted pump; C shows systolic blood pressure in mice injected with WRR139 or a vehicle control, given saline or angiotensin II via a subcutaneous implanted pump; and D shows diastolic blood pressure in mice injected with WRR139 or a vehicle control, given saline or angiotensin II via a subcutaneous implanted pump. The results showed that pre-injection of the Nrf3 inhibitors nelfinavir or WRR139 significantly reduced systolic blood pressure (P < 0.05) and diastolic blood pressure (P < 0.01) in mice with angiotensin II-induced hypertension, compared with the control group injected with a vehicle control.

[0058] 3. Secondary mesenteric artery diastolic function test

[0059] After the mouse was anesthetized, the secondary mesenteric artery was quickly removed and transferred to ice-cold Krebs solution. After removing the fat around the blood vessels, the blood vessels were cut into pieces of about 2 to 3 mm in length, suspended in the Multi Myograph System detection system, and Krebs-Ringer bicarbonate solution containing 5% CO2 and 95% O2 was added. After each blood vessel was balanced in the solution for 30 minutes, it was allowed to reach a stable resting tension. Then 60mmol / LKCl was added to each blood vessel to detect its contractile function to determine the activity of the blood vessel after ex vivo. After confirming that the vascular activity was normal, phenylephrine (PE, 10 -5 After inducing vascular precontraction, acetylcholine (Ach, 10 -9 , 10 -8 , 10 -7 , 10 -6 , 10 -5 mol / L) to dilate blood vessels, thereby detecting the endothelium-dependent vasodilation function of blood vessels.

[0060] The results are as follows Figure 4As shown in the figure, A shows the mesenteric artery relaxation function of mice injected with nelfinavir or solvent control in mice given saline or angiotensin II by subcutaneous implanted pump; B shows the mesenteric artery relaxation function of mice injected with WRR139 or solvent control in mice given saline or angiotensin II by subcutaneous implanted pump; the results showed that the injection of Nrf3 inhibitors nelfinavir or WRR139 can significantly inhibit the angiotensin II-induced mesenteric artery relaxation dysfunction in mice (P < 0.05).

[0061] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. Use of Nrf3 inhibitors in the preparation of drugs for preventing and / or treating hypertension.

2. The use according to claim 1, characterized in that The inhibition rate of the Nrf3 inhibitor on the biological activity of Nrf3 is 50% to 100%.

3. The use according to claim 1, characterized in that The Nrf3 inhibitors include HIV-1 protease inhibitors and / or peptide vinyl sulfone compounds.

4. The use according to claim 3, characterized in that The HIV-1 protease inhibitors include any one or more of ritonavir, nelfinavir and saquinavir.

5. The use according to claim 3, characterized in that The peptide vinyl sulfone compounds include WRR139.

6. The use according to claim 1, characterized in that The Nrf3 inhibitor alleviates arterial diastolic dysfunction caused by hypertension.

7. The use according to claim 6, characterized in that The hypertension is angiotensin II-induced hypertension.

8. The use according to claim 1, characterized in that The active ingredient of the drug includes an Nrf3 inhibitor.

9. The use according to claim 8, characterized in that The drug also includes pharmaceutically acceptable excipients.

10. The use according to claim 8, characterized in that The dosage form of the drug includes any one or more of oral preparations, intravenous injections and intramuscular injections.

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