Phactr1 transrepressor inhibitor ccc-1423 for the treatment of hypertension

Abstract

The application belongs to the field of cardiovascular disease treatment, and provides application of a Phactr1 nuclear transfer inhibitor in hypertension treatment. Specifically, the Phactr1 nuclear transfer inhibitor CCG-1423 restores endothelial function and reduces hypertension by inhibiting Phactr1 nuclear transfer.

Description

Technical Field

[0001] This invention belongs to the field of cardiovascular disease treatment, specifically involving the Phactr1 nuclear transfer inhibitor CCG-1423 for the treatment of hypertension. Background Technology

[0002] Hypertension, also known as high blood pressure, is a condition where the pressure exerted by blood on the walls of blood vessels is consistently higher than normal. According to the "Chinese Clinical Practice Guidelines for Hypertension," the diagnostic criteria for hypertension in adults are a systolic blood pressure ≥130 mmHg and / or a diastolic blood pressure ≥80 mmHg.

[0003] The causes of hypertension are mostly genetic factors and unhealthy lifestyles, such as high-salt diets, excessive alcohol consumption, chronic stress, and insufficient physical activity. Hypertension is more prevalent in five groups: those with a family history of hypertension; those who are easily agitated; those with high salt intake; those who consume excessive alcohol; and those experiencing high work or life stress.

[0004] Typical symptoms of hypertension include headache, fatigue or restlessness, arrhythmia, palpitations, and tinnitus. However, many people with hypertension may develop other complications without any symptoms, such as stroke, blurred vision, loss of consciousness, and memory loss. Therefore, hypertension is known as the "silent killer."

[0005] For patients with hypertension, lifestyle modifications and medication are the most common treatment methods. The basic principles of medication include prioritizing long-acting antihypertensive drugs to control blood pressure over 24 hours, and adjusting the medication and dosage as needed. If a patient's blood pressure remains uncontrolled for a long period, it can lead to more serious complications, and even potentially fatal outcomes such as stroke or cerebral infarction.

[0006] A diagnosis of hypertension involves multiple assessments and examinations, primarily including observation of clinical symptoms, blood pressure measurement, and related auxiliary examinations such as electrocardiogram (ECG) and kidney function tests. However, some people are still diagnosed with hypertension and start taking antihypertensive medications simply by measuring their blood pressure. Nevertheless, existing antihypertensive drugs can lead to drug resistance and have side effects on healthy individuals.

[0007] Therefore, there is an urgent need to provide a class of therapeutic drugs that can effectively improve blood pressure problems and endothelial dysfunction in hypertensive patients, while having no obvious side effects on healthy people. Summary of the Invention

[0008] This invention aims to provide the application of Phactr1 nuclear transport inhibitors in restoring endothelial dysfunction, thereby treating or improving hypertension.

[0009] In a first aspect of the invention, there is provided the use of a Phactr1 nuclear transport inhibitor for preparing a formulation or composition for one or more of the following uses:

[0010] (Z0) Treatment of hypertension in the subjects;

[0011] (Z1) improves endothelial dysfunction;

[0012] (Z2) improves vasodilatory function;

[0013] (Z3) Increases the expression of endothelial nitric oxide synthase eNOS;

[0014] (Z4) increases NO secretion.

[0015] In another preferred embodiment, the nuclear transport inhibitor of Phactr1 is CCG-1423.

[0016] In another preferred embodiment, the treatment of hypertension in the subject includes: lowering the subject's systolic blood pressure, lowering the subject's diastolic blood pressure, maintaining the subject's blood pressure stability, or a combination thereof.

[0017] In another preferred embodiment, the subject is a person diagnosed with hypertension.

[0018] In another preferred embodiment, the subject does not have diabetes.

[0019] In another preferred embodiment, the formulation or composition does not lower the blood pressure of a normal control.

[0020] In another preferred embodiment, the subject has one or more of the following characteristics:

[0021] (a) Endothelial dysfunction;

[0022] (b) The expression level of endothelial nitric oxide synthase eNOS was significantly lower than that of the normal control;

[0023] (c) NO secretion levels were significantly lower than those of the normal control.

[0024] In another preferred embodiment, the endothelial dysfunction includes: vasodilatory dysfunction, blood flow impairment, substance transport dysfunction, or a combination thereof.

[0025] In another preferred embodiment, the method for determining vascular endothelial dysfunction includes: coronary angiography, direct Doppler wire measurement, positron emission tomography, myocardial perfusion imaging, oxygen level-dependent magnetic resonance imaging, echocardiography, brachial artery flow-mediated vasodilatory function (FMD) detection, peripheral arterial tension (PAT) detection, or a combination thereof.

[0026] In another preferred embodiment, the ratio A1 / A0 of the expression level A1 of the endothelial nitric oxide synthase (eNOS) in the test subject and the expression level A0 of eNOS in the normal control is ≤0.8, preferably ≤0.6, and most preferably ≤0.5; and / or

[0027] The ratio of the NO secretion level B1 of the test subjects to the NO secretion level B0 of the normal controls, B1 / B0, is ≤0.8, preferably ≤0.6, and most preferably ≤0.5.

[0028] In another preferred embodiment, the ratio B2 / B1 of the NO secretion level B2 of the test subject after administration of the formulation or composition and the NO secretion level B1 of the test subject before administration of the formulation or composition is ≥30%, preferably ≥40%, and most preferably ≥50%.

[0029] In another preferred embodiment, the ratio A2 / A1 of the expression level A2 of endothelial nitric oxide synthase (eNOS) in the test subject after administration of the formulation or composition to the expression level A1 of endothelial nitric oxide synthase (eNOS) in the test subject before administration of the formulation or composition is ≥30%, preferably ≥50%, and most preferably ≥60%.

[0030] In another preferred embodiment, the NO secretion includes endothelial NO secretion and / or plasma NO secretion.

[0031] In a second aspect of the invention, a pharmaceutical composition for treating hypertension is provided, the pharmaceutical composition comprising CCG-1423.

[0032] In a third aspect of the invention, a pharmaceutical combination for treating hypertension is provided, the pharmaceutical combination comprising a first pharmaceutical component and a second pharmaceutical component, the first pharmaceutical component being CCG-1423 and the second pharmaceutical component being other hypertension treatment drugs.

[0033] In another preferred embodiment, the second pharmaceutical component includes: dihydropyridine drugs, ACE inhibitors, ARBs, diuretics, beta-blockers, PP1α inhibitors, Sp1 agonists, or combinations thereof.

[0034] In a fourth aspect of the invention, the use of the pharmaceutical composition described in the second aspect of the invention or the pharmaceutical composition described in the third aspect of the invention is provided for preparing a medicine box for treating hypertension.

[0035] In another preferred embodiment, the medicine box is also used for the following purposes:

[0036] (Z1) improves endothelial dysfunction;

[0037] (Z2) improves vasodilatory function;

[0038] (Z3) Increases the expression of endothelial nitric oxide synthase eNOS; and / or

[0039] (Z4) increases NO secretion.

[0040] In another preferred embodiment, the NO secretion includes endothelial NO secretion and / or plasma NO secretion.

[0041] In a fifth aspect of the invention, a medicine box for treating hypertension is provided, the medicine box comprising the following components:

[0042] (i) the pharmaceutical composition described in the second aspect of the present invention or the pharmaceutical composition described in the third aspect of the present invention;

[0043] (ii) Optional detection reagents for detecting NO secreted in vascular endothelium and / or plasma.

[0044] In a sixth aspect of the invention, a method for screening potential therapeutic agents for treating hypertension is provided, comprising the following steps:

[0045] (S1) In the experimental group, aortic endothelial cells (HAECs) were cultured in the presence of the test substance; and in the control group, aortic endothelial cells (HAECs) were cultured under the same conditions but in the absence of the test substance.

[0046] (S2) The Phactr1 nuclear transport level of HAECs in the experimental group and the control group was measured and compared. If the Phactr1 nuclear transport level Y1 in the experimental group was significantly reduced compared with the Phactr1 nuclear transport level Y0 in the control group, it suggests that the test substance is a potential therapeutic agent for the treatment of hypertension.

[0047] In another preferred embodiment, the hypertension is endothelial dysfunction hypertension.

[0048] In another preferred embodiment, the test substance is a single compound, a mixture, an extract, or a compound preparation.

[0049] In another preferred embodiment, the method further includes: (S3) further performing activity testing or functional testing on the potential therapeutic agents selected in the previous step.

[0050] In another preferred embodiment, “significant reduction” means Y1 / Y0 ≤ 0.8, more preferably ≤ 0.7, and even more preferably ≤ 0.6.

[0051] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0052] Figure 1 The structural diagram of CCG-1423 is shown.

[0053] Figure 2 The results showed that CCG1423 treatment of HAEC significantly inhibited PHACTR1 nuclear translocation and restored eNOS expression and NO secretion.

[0054] in, Figure 2 A shows the Western blot results of CCG1423 expression in the cell nucleus and cytoplasm; Figure 2 B shows the effect of CCG1423 on the mRNA level of eNOS expression; Figure 2 C showed that CCG1423 promotes the secretion of NO by endothelial cells.

[0055] Figure 3 The study showed that CCG-1423 treatment in hypertensive mice significantly reduced blood pressure, restored eNOS levels and NO secretion, and restored endothelial diastolic function.

[0056] in, Figure 3 A shows the results of systolic blood pressure changes assessed using the tail sleeve method; Figure 3 B shows the results of systolic blood pressure changes assessed by the telemetry system; Figure 3 C shows the results of diastolic blood pressure changes assessed by the telemetry system; Figure 3 D shows the results of the vascular ring experiment; Figure 3 E shows that CCG-1423 upregulates eNOS expression; Figure 3 F shows that CCG-1423 promotes in situ NO secretion in the vascular wall; Figure 3 G indicates the secretion of NO in the plasma. Detailed Implementation

[0057] Through extensive and in-depth research, the inventors unexpectedly discovered for the first time that the Phactr1 nuclear transport inhibitor CCG-1423 can restore endothelial function in hypertensive subjects, upregulate eNOS expression, and promote nitric oxide (NO) secretion in hypertensive subjects by blocking the Phactr1 / PP1α / Sp1 signaling pathway, thereby playing a therapeutic role in hypertension. Simultaneously, the Phactr1 nuclear transport inhibitor CCG-1423 has almost no effect on blood pressure in healthy subjects. Based on this, the present invention was completed.

[0058] Specifically, this invention uses hypertensive model mice and normal C57BL / 6J mice to conduct multiple in vitro and in vivo tests, including blood pressure monitoring, vascular function testing, eNOS expression and NO secretion level measurement, to evaluate how CCG-1423 restores endothelial function, reduces blood pressure, and improves vasodilatory function by inhibiting Phactr1 nuclear transport.

[0059] the term

[0060] To facilitate a clearer understanding of this disclosure, certain terms are first defined. As used herein, unless otherwise expressly specified herein, each of the following terms shall have the meaning given below. Other definitions are set forth throughout the application.

[0061] As used herein, the term “and / or” refers to and covers any and all possible combinations of one or more of the related listed items.

[0062] As used herein, the terms “comprising,” “including,” and “containing” are used interchangeably and include not only closed definitions but also semi-closed and open definitions. In other words, the terms include “consisting of” and “substantially consisting of”.

[0063] Where a numerical range is provided, unless the context clearly indicates otherwise, it should be understood that every intermediate integer of the value 20, every tenth of every intermediate integer of the value, any other intermediate value between the upper and lower limits of the range, and any other intermediate value within the specified range are included in this invention. The upper and lower limits of these smaller ranges may be independently included within the smaller range and also covered by this invention, but are subject to any express exclusions within the specified range. For example, "1 to 50" includes "2 to 25", "5 to 20", "25 to 50", "1 to 10", etc.

[0064] hypertension

[0065] Hypertension is one of the most common chronic diseases worldwide and a significant risk factor for cardiovascular disease. Its pathogenesis is complex, involving multiple pathological processes such as vascular endothelial dysfunction, vascular remodeling, and neuroendocrine disorders. Among these, vascular endothelial dysfunction is a key pathological link in the occurrence and development of hypertension, manifested as reduced nitric oxide (NO) production, enhanced vasoconstriction, exacerbated oxidative stress, and escalated inflammatory responses.

[0066] Endothelial nitric oxide synthase (eNOS) is the main enzyme responsible for NO production in endothelial cells, and its activity is crucial for maintaining vascular function. Under normal conditions, eNOS is activated by phosphorylation through signaling pathways such as Akt and AMPK, thereby generating NO to maintain vasodilation. However, in hypertensive states, eNOS expression and activity are significantly reduced, leading to insufficient NO production, which further promotes vasoconstriction and increased blood pressure. In addition, oxidative stress in hypertensive states leads to excessive production of reactive oxygen species (ROS) mediated by NADPH oxidase, which not only consumes NO but also activates pro-inflammatory signaling pathways (such as NF-κB and MAPK), exacerbating vascular damage.

[0067] Existing antihypertensive treatment options include traditional ACE inhibitors, Ang II receptor antagonists (ARBs), and antioxidant therapy. These drugs have shown some effectiveness in improving blood pressure control, but their direct improvement on endothelial dysfunction is limited, and some treatments have issues such as poor tolerability and side effects. Furthermore, while novel drug delivery systems (such as exosomes and nanoparticles) have shown potential for strong targeting and high bioavailability, their safety and long-term efficacy require further validation.

[0068] Treatment options for hypertension include therapies targeting the endothelial nitric oxide (NO) pathway, therapies inhibiting oxidative stress, therapies blocking the renin-angiotensin system (RAS), therapies targeting endothelial inflammation, and nanoparticle and exosome therapies.

[0069] Targeting the endothelial nitric oxide (NO) pathway. NO produced by endothelial nitric oxide synthase (eNOS) is a key molecule for maintaining vasodilation. In hypertension, decreased eNOS expression and activity lead to insufficient NO production. Some studies have attempted to improve vascular function by increasing NO bioavailability (e.g., NO donors) or activating eNOS-related pathways (e.g., AMPK and Akt). However, these therapeutic strategies still face challenges such as limited NO bioavailability and risks of side effects (e.g., hypotension and oxidative stress).

[0070] Treatment aimed at inhibiting oxidative stress. Oxidative stress is one of the main causes of endothelial dysfunction in hypertension, mediated by excessive NADPH oxidase and reactive oxygen species (ROS). Oxidative stress not only reduces the bioavailability of NO but also activates pro-inflammatory signals (such as NF-κB and MAPK), exacerbating vascular damage. Antioxidants or drugs that inhibit NADPH oxidase activity have shown potential efficacy, but their clinical effectiveness is limited, mainly because the oxidative stress regulatory network is complex and global inhibition may affect normal cellular signaling.

[0071] Treatment involves blocking the renin-angiotensin system (RAS). Overactivation of the RAS through blocking the renin-angiotensin system promotes vasoconstriction, oxidative stress, and inflammatory responses via angiotensin II (Ang II). ACE inhibitors (such as captopril) and Ang II receptor antagonists (ARBs, such as losartan) are currently the most commonly used antihypertensive drugs in clinical practice. These drugs improve endothelial function by reducing Ang II production or blocking its effects. However, some patients may have poor tolerance to RAS inhibitors.

[0072] Targeted therapy for endothelial inflammation. Endothelial inflammation in hypertension is mediated by various pro-inflammatory factors (such as TNF-α and IL-6) and cell adhesion molecules (such as ICAM-1 and VCAM-1), further promoting vascular remodeling and sclerosis. Studies have shown that biologics targeting inflammatory factors (such as IL-6 antagonists) may have potential in reducing vascular inflammation and improving endothelial function, but their widespread application still requires more evidence.

[0073] Nanoparticle and exosome therapy. Nanoparticles and exosomes, as drug carriers, can improve drug targeting and bioavailability. Studies have shown that exosomes containing microRNAs or anti-inflammatory factors can improve vascular function by regulating endothelial function-related signaling pathways (such as eNOS and ROS). These novel technologies offer promising directions for the future treatment of hypertension, but their safety and long-term efficacy still need to be verified.

[0074] However, existing antihypertensive drugs often suffer from insufficient targeting, difficulty in comprehensively regulating oxidative stress and inflammation, and remaining technological bottlenecks. For example, existing antihypertensive drugs (such as ACE inhibitors and ARBs) mainly act on the renin-angiotensin system (RAS), effectively controlling hypertension overall, but their direct improvement on vascular endothelial dysfunction is limited, especially for endothelial dysfunction caused by insufficient eNOS expression and reduced NO production, where current treatments lack specific targets. Although antioxidants and inflammation inhibitors have shown potential to improve endothelial function in experimental studies, oxidative stress and inflammation are complex biological networks, and non-specific inhibition may lead to physiological signal disturbances or side effects, limiting clinical application. Novel drug delivery systems such as nanoparticles and exosomes have the advantages of strong targeting and high delivery efficiency, but their preparation costs are high, drug loading efficiency is insufficient, and the safety of long-term use requires further investigation.

[0075] Phactr1 inhibitors

[0076] Phactr1 inhibitors are a class of substances used to target and regulate the activity of protein phosphatases and actin regulatory protein 1 (Phactr1).

[0077] Phactr1 is a protein that plays a crucial role in cellular processes involving actin dynamics and signal transduction pathways. Phactr1 inhibitors are primarily used for research purposes to gain a deeper understanding of Phactr1's function in the cellular environment. The Phactr1 inhibitor of this invention aims to inhibit Phactr1 activity, ultimately altering the phosphorylation patterns of its target proteins.

[0078] The Phactr1 inhibitors of the present invention comprise reagents or compositions capable of interfering with Phactr1-related regulatory pathways. Some of these inhibitors (such as calpain inhibitors and okadaic acid) act indirectly by targeting protein phosphatases (PP1 and PP2A) that interact with Phactr1. By inhibiting these phosphatases, they prevent the dephosphorylation of Phactr1-targeting proteins, thereby altering their phosphorylation state and disrupting cellular signaling cascades. Other drugs, such as cyclosporine A and FK506, act indirectly by binding to specific proteins (cyclosporine protein and FKBP12, respectively) and inhibiting calmoneurin phosphatase (a phosphatase involved in Phactr1-mediated processes).

[0079] In specific embodiments, the dosage forms of the Phactr1 inhibitor of the present invention include: solid dosage forms, liquid dosage forms, and semi-solid dosage forms. In specific embodiments, the Phactr1 inhibitor of the present invention can be any formulation that inhibits Phactr1 nuclear transport, including (but not limited to): nucleic acids, proteins, peptides, small molecules, and complex substances.

[0080] CCG-1423

[0081] CCG-1423 is an inhibitor of the Rho / MRTF / SRF pathway. CCG-1423 is active in various cancer cells. CCG-1423 is a lead compound for developing novel bioactive molecular tools that can be used in cancer and diabetes research. Its molecular formula is as follows: Figure 1 As shown.

[0082] Uses of the present invention

[0083] This invention provides new uses for CCG-1423, mainly including one or more of the following uses:

[0084] As a small molecule inhibitor, CCG-1423 specifically inhibits the transport of Phactr1 from the cytoplasm to the nucleus, thereby regulating the Phactr1 / PP1α / Sp1 signaling pathway, restoring eNOS expression and NO production, significantly improving endothelial function, and reducing blood pressure.

[0085] Specific antihypertensive effect: Specifically, CCG-1423 showed a significant antihypertensive effect in hypertensive mice, while having no significant effect on normal C57BL / 6J mice, demonstrating its targeted and specific effect on hypertension.

[0086] Specifically, CCG-1423 significantly improved vasodilation function in hypertensive mice, particularly by enhancing the vasodilatory response of the aorta to acetylcholine and restoring endothelium-dependent vasodilation function.

[0087] By enhancing NO production through eNOS regulation, CCG-1423 specifically addresses endothelial dysfunction caused by insufficient eNOS expression and reduced NO production by upregulating eNOS expression and enhancing NO secretion.

[0088] The method of the present invention

[0089] This invention addresses the shortcomings of existing technologies in treating hypertension, such as insufficient improvement of endothelial dysfunction, lack of targeting, and inadequate intervention at the molecular level. It proposes a more promising treatment method for hypertension, specifically by targeting and inhibiting Phactr1 nuclear transport with CCG-1423, thereby solving the following technical problems:

[0090] This invention aims to improve hypertension-related endothelial dysfunction. Existing treatments are unable to effectively reverse endothelial dysfunction, particularly vascular dysfunction caused by insufficient eNOS expression and reduced NO production. This invention addresses this issue by blocking Phactr1 nuclear transport, restoring Sp1 phosphorylation levels, promoting eNOS transcription, and significantly increasing NO production, thereby fundamentally improving endothelial-dependent vasodilation.

[0091] Improving the targeting of treatment. Addressing the lack of specificity in antioxidants and anti-inflammatory drugs, this invention focuses on the Phactr1 / PP1α / Sp1 signaling pathway, a specific regulatory axis, to precisely intervene in the molecular mechanisms related to endothelial dysfunction, avoiding interference with normal physiological processes by traditional drugs.

[0092] This invention fills a gap in molecular mechanism intervention. Existing treatments have failed to address the key role of Phactr1 nuclear transport. This invention utilizes CCG-1423, a small molecule inhibitor, to directly block Phactr1 nuclear transport, achieving for the first time precise regulation of the Phactr1 / PP1α / Sp1 signaling pathway, thus filling a technological gap in the treatment of endothelial dysfunction.

[0093] The method of this invention solves the core problem of insufficient intervention in hypertension endothelial dysfunction in existing technologies, and provides an innovative and more precise treatment method for hypertension.

[0094] In addition to CCG-1423, more specific Phactr1 nuclear transport inhibitors can be designed by optimizing the molecular structure or introducing new targeting strategies to improve the precise inhibition of Phactr1 nuclear transport. Alternatively, Phactr1 nuclear transport inhibitors with higher targeting and fewer potential side effects can be obtained directly through methods such as screening compound libraries and structural optimization for effective and precise regulation of hypertension.

[0095] In addition to inhibiting Phactr1 translocation, other drugs targeting the Phactr1 / PP1α / Sp1 signaling pathway can be explored. For example, inhibitors of PP1α or agonists of Sp1 can further restore eNOS expression and improve NO production by modulating the activity of these molecules upstream or downstream. These alternative drugs can be used as complementary therapies in combination with CCG-1423 to enhance therapeutic efficacy and specificity.

[0096] The main advantages of this invention include:

[0097] (a) This invention provides the application of Phactr1 nuclear transport inhibitors in effectively reversing endothelial dysfunction, particularly vascular dysfunction caused by insufficient eNOS expression and reduced NO production. Specifically, it promotes eNOS transcription, significantly increases NO production, and fundamentally improves endothelial-dependent vasodilation.

[0098] (b) The nuclear transport inhibitor of Phactr1, CCG-1423, has high targeting specificity and has no significant effect on normotensive mice.

[0099] (c) This invention provides a method for lowering blood pressure by using CCG-1423 to block Phactr1 nuclear transport, restore the phosphorylation level of Sp1, achieve precise regulation of the Phactr1 / PP1α / Sp1 signaling pathway, improve endothelial function, restore NO production, and thus lower blood pressure.

[0100] (d) The blood pressure reduction method provided by the present invention avoids the side effects that may be caused by systemic oxidative stress and inflammatory suppression, and provides a safer and more efficient treatment strategy with significant clinical application potential.

[0101] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0102] Example 1: CCG-1423 promotes the expression of eNOS in endothelial cells.

[0103] In this embodiment, aortic endothelial (HAECs) cells (1x10⁻¹) were treated with CCG-1423 (200 nM). 6 (1), to observe its effects on Phactr1 nuclear transport, eNOS expression and NO secretion.

[0104] The effects of CCG-1423 on Phactr1 nuclear transport are as follows: Figure 2 As shown in Figure A, the results indicate that CCG-1423 treatment significantly inhibited the nuclear transport of Phactr1, with most Phactr1 remaining in the cytoplasm, suggesting that CCG-1423 can effectively inhibit the nuclear transport of Phactr1.

[0105] The effect of CCG-1423 on eNOS expression is as follows: Figure 2 As shown in Figure B, HTN represents HAECs treated with Heg1-shRNA, where Heg1 knockout leads to downregulation of eNOS, thus simulating a hypertension model; Vehicle is DMSO, representing the CCG-1423 control group. Specifically, the Control+Vehicle group represents HAECs treated with Scramble-shRNA plus DMSO, representing healthy cells without CCG-1423; the Control+CCG-1423 group represents HAECs treated with Scramble-shRNA plus CCG-1423, representing healthy cells with CCG-1423; the HTN+Vehicle group represents hypertensive HAECs without CCG-1423; and the HTN+CCG-1423 group represents hypertensive HAECs with CCG-1423. The results show that eNOS expression increased by approximately 50% after CCG-1423 treatment.

[0106] The effect of CCG-1423 on NO secretion, such as Figure 2As shown in Figure C, the results indicate that NO production levels increased by approximately 60%, confirming that CCG-1423 promotes the expression of eNOS in endothelial cells.

[0107] Example 2: CCG-1423 promotes the expression of eNOS in endothelial cells.

[0108] In this embodiment, C57 normal mice and hypertensive model mice were administered CCG-1423 (1 mg / kg) via intraperitoneal injection daily. Blood pressure was monitored using the tail sleeve method and telemetry system. Vascular dilation function was assessed using the vascular ring experiment, and eNOS expression and NO secretion were detected.

[0109] The results of systolic blood pressure monitoring using the tail sleeve method are as follows: Figure 3 As shown in Figure A, at week 20, systolic and diastolic blood pressure were monitored in live mice using a telemetry system. The results are as follows: Figure 3 B and Figure 3 As shown in C.

[0110] The results showed that, since the mouse model of hypertension was induced by endothelial-specific knockout of Heg1, the increase in blood pressure was gradual. Subsequently, in the hypertensive model mice, the blood pressure of the CCG-1423-treated hypertensive model mice (experimental group mice) was significantly reduced, with systolic blood pressure (SBP) and diastolic blood pressure (DBP) decreasing by approximately 20-40 mmHg. Specifically, the SBP of the hypertensive mice decreased from 145±5 mmHg to 110±4 mmHg, and the DBP decreased from 105±4 mmHg to 85±3 mmHg. This result indicates that CCG-1423 can effectively reduce hypertension.

[0111] Furthermore, the results showed that CCG-1423 did not have a significant antihypertensive effect when applied to normal C57BL / 6J mice. SBP and DBP in normal mice did not change significantly before and after CCG-1423 treatment. This indicates that CCG-1423 has no effect on normal blood pressure, further demonstrating its specificity in antihypertensive treatment, producing a significant effect only on hypertensive model mice.

[0112] Results of the vascular ring experiment are as follows Figure 3 As shown in Figure D, CCG-1423 treatment significantly improved vascular dilation function in hypertensive mice. Vascular ring experiments revealed a significantly enhanced vasodilatory response of the aorta to acetylcholine in the experimental group mice, with the amplitude of dilation increasing by approximately 30% compared to the control group. This indicates that CCG-1423 can restore vascular dilation capacity and improve vascular function in hypertensive mice.

[0113] eNOS expression results are as follows Figure 3 As shown in E, the NO secretion results are as follows: Figure 3 F and Figure 3 As shown in G, where... Figure 3F represents the results of in-situ NO staining of the blood vessel wall. Figure 3 G is the serum NO level detected using the Griess reaction method.

[0114] The results showed that in in vivo experiments, the expression of eNOS protein in the vascular tissue of CCG-1423-treated hypertensive mice increased significantly in approximately 50% of cases, and NO secretion levels increased significantly in approximately 60% of cases. Serum NO levels, detected by the Griess reaction method, showed a significant increase in NO concentration in the experimental group mice. This further validated that CCG-1423 increases NO synthesis by upregulating eNOS expression, thereby improving endothelial function and vasodilation.

[0115] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. The use of a Phactr1 nuclear transport inhibitor, characterized in that, For use in the preparation of a formulation or composition, said formulation or composition for one or more of the following purposes: (Z0) Treatment of hypertension in the subjects; (Z1) improves endothelial dysfunction; (Z2) improves vasodilatory function; (Z3) Increases the expression of endothelial nitric oxide synthase eNOS; (Z4) increases NO secretion.

2. The use as described in claim 1, characterized in that, The nuclear transport inhibitor of Phactr1 is CCG-1423.

3. The use as described in claim 1, characterized in that, The test subjects have one or more of the following characteristics: (a) Endothelial dysfunction; (b) The expression level of endothelial nitric oxide synthase eNOS was significantly lower than that of the normal control; (c) NO secretion levels were significantly lower than those of the normal control.

4. The use as described in claim 1, characterized in that, The ratio of the expression level A1 of endothelial nitric oxide synthase (eNOS) in the test subjects to the expression level A0 of eNOS in the normal controls, A1 / A0, is ≤0.8, preferably ≤0.6, and most preferably ≤0.5; and / or The ratio of the NO secretion level B1 of the test subjects to the NO secretion level B0 of the normal controls, B1 / B0, is ≤0.8, preferably ≤0.6, and most preferably ≤0.

5.

5. A pharmaceutical composition for treating hypertension, characterized in that, The pharmaceutical composition contains CCG-1423.

6. A pharmaceutical combination for treating hypertension, characterized in that, The drug combination includes a first drug component and a second drug component, wherein the first drug component is CCG-1423 and the second drug component is other hypertension treatment drugs.

7. The pharmaceutical combination as described in claim 6, characterized in that, The second drug component includes: dihydropyridine calcium channel blockers, ACE inhibitors, sartans, diuretics, beta-blockers, PP1α inhibitors, Sp1 agonists, or combinations thereof.

8. The use of the pharmaceutical composition of claim 5 or the pharmaceutical composition of claim 6, characterized in that, This is used to prepare a medicine box for treating hypertension.

9. A medicine box for treating hypertension, characterized in that, The medicine box contains the following components: (i) the pharmaceutical composition of claim 5 or the pharmaceutical composition of claim 6; (ii) Optional detection reagents for detecting NO secreted in vascular endothelium and / or plasma.

10. A method for screening potential therapeutic agents for treating hypertension, characterized in that, Includes the following steps: (S1) In the experimental group, aortic endothelial cells (HAECs) were cultured in the presence of the test substance; and in the control group, aortic endothelial cells (HAECs) were cultured under the same conditions but in the absence of the test substance. (S2) The Phactr1 nuclear transport level of HAECs in the experimental group and the control group was measured and compared. If the Phactr1 nuclear transport level Y1 in the experimental group was significantly reduced compared with the Phactr1 nuclear transport level Y0 in the control group, it suggests that the test substance is a potential therapeutic agent for the treatment of hypertension.

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