Use of cyclin-dependent kinase inhibitors in the treatment of heart failure with preserved ejection fraction

Abstract

The present invention relates to the use of a cyclin-dependent kinase inhibitor for the treatment of heart failure with preserved ejection fraction. The use is in particular the use of the cyclin-dependent kinase (CDK4 / 6) inhibitor palbociclib for the prevention and / or treatment of heart failure with preserved ejection fraction by oral administration.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of cyclin-dependent kinase inhibitors in the treatment of heart failure with preserved ejection fraction. Background Technology

[0002] Heart failure with preserved ejection fraction (HFpEF) is the most common type of heart failure in my country and globally, defined as heart failure with a left ventricular ejection fraction (LVEF) ≥ 50%. HFpEF is a complex cardiovascular syndrome characterized by left ventricular hypertrophy, concentric remodeling, increased extracellular matrix, abnormal calcium processing, diastolic and perfusion abnormalities, and decreased diastolic elasticity. HFpEF has a high morbidity and mortality rate, and patients are typically older, predominantly female, and often have comorbidities such as hypertension, atrial fibrillation, diabetes, kidney disease, and obesity, resulting in significant clinical heterogeneity.

[0003] The pathophysiological mechanisms of HFpEF are complex, involving multiple organ dysfunctions and interactions between multiple pathways, thus it has always been a challenge in the treatment of heart failure.

[0004] Due to the complexity of the pathophysiological mechanisms of HFpEF and its interaction with common related comorbidities, treatment of HFpEF should begin with addressing risk factors and comorbidities. Commonly used drugs for the pharmacological treatment of HFpEF include beta-blockers, aldosterone receptor antagonists, angiotensin-converting enzyme inhibitors / angiotensin II receptor antagonists, and sodium-glucose cotransporter 2 inhibitors (SGLT2i).

[0005] in,

[0006] β-blockers are commonly used to treat comorbidities such as coronary artery disease and atrial fibrillation. A meta-analysis of three medium-sized randomized clinical trials showed that β-blockers can reduce all-cause mortality in patients with heart failure with paroxysmal pulmonary embolism (HFpEF). However, while β-blocker treatment is beneficial in some patients, it can worsen the disease in others, and its efficacy in HFpEF requires further confirmation through more randomized clinical trials. The roles of renin-angiotensin-aldosterone system inhibitors and mineralocorticoid receptor antagonists in heart failure with paroxysmal pulmonary embolism (HFrEF) are well-established, but their effects on HFpEF are not significant, possibly due to the lower importance of RAAS system abnormalities in the pathophysiological mechanisms of HFpEF. Clinical trials of angiotensin-converting enzyme inhibitors and angiotensin II receptor antagonists have not significantly reduced all-cause mortality or cardiovascular mortality risk in HFpEF. In the EMPEROR-Preserved, DELIVER, and SOLOIST-WHFDELIVER clinical trials, SGLT2 inhibitor empagliflozin reduced the risk of combined cardiovascular death or hospitalization for heart failure in heart failure patients with LVEF > 40%. The 2022 ESC Heart Failure Guidelines and the 2022 AHA / ACC / HFSA Guidelines list SGLT2i as a Class 2a recommendation for treating patients with heart failure with reduced ejection fraction (HFpEF). However, in these clinical trials, only a portion of the enrolled patients had an ejection fraction ≥50%, and the mortality-reducing effect of the drug diminished with increasing left ventricular ejection fraction (LVEF), indicating that SGLT2i is only effective for a subset of HFpEF patients. Furthermore, SGLT2i also has many adverse reactions and side effects in clinical practice, such as ketoacidosis, volume depletion, and reversible mild elevation of serum creatinine. Therefore, exploring safer and more effective drugs to prevent and alleviate HFpEF is crucial.

[0007] Based on this, the present invention is proposed. Summary of the Invention

[0008] This invention relates first to the use of the cyclin-dependent kinase (CDK4 / 6) inhibitor palbociclib in the preparation of medicaments and / or pharmaceutical compositions for the treatment and / or prevention of heart failure with preserved ejection fraction.

[0009] Furthermore, the drug is: a drug administered via the gastrointestinal tract, a drug administered via intravenous injection, or a drug administered via subcutaneous implantation, preferably a drug administered via the gastrointestinal tract.

[0010] Furthermore, the drug and / or pharmaceutical composition contains a therapeutically effective amount of palbociclib, as well as necessary pharmaceutical excipients.

[0011] In normal myocardium, energy metabolism is flexible, with fatty acid β-oxidation, glucose oxidation, and glycolysis in a dynamic equilibrium to effectively maintain energy supply to cardiomyocytes. However, in heart failure, this dynamic equilibrium is disrupted, and end-stage cardiomyocytes shift from primarily relying on fatty acid metabolism to primarily relying on glucose metabolism for energy. Cyclin-dependent kinases (CDKs) and cyclins are core molecules in the entire cell cycle regulation mechanism. Cyclin D3-CDK6 kinase can phosphorylate two key enzymes in glucose metabolism, phosphofructokinase 6-phosphate and M2 pyruvate kinase, and inhibit their metabolic activity, directly activating the pentose phosphate pathway and serine pathway in glucose metabolism. Protein kinases, as important signaling messengers in cellular life activities, are key regulators for achieving cellular function and regulating cellular metabolism. Protein kinase regulators are also a relatively new class of drugs that can regulate the cell cycle, control cardiac metabolic function, and regulate transcription levels.

[0012] The beneficial effects of this invention are as follows:

[0013] By using animal models to explore the pathological mechanism of HFpEF, a new drug, the cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor Palbociclib, was discovered that can treat HFpEF.

[0014] In summary, this application reports for the first time the therapeutic effect of the cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor palbociclib on heart failure with preserved ejection fraction. Attached Figure Description

[0015] Figure 1 The therapeutic effect of CDK4 / 6 inhibitor (Palbociclib) on a mouse model of heart failure with preserved ejection fraction.

[0016] Figure 2 The structural formula of palbociclib.

[0017] Figure 3 1. Pathological staining results of cardiac adipose tissue (HE staining) in model mice before and after treatment. Detailed Implementation

[0018] Biochemical reagents and kits

[0019] Table 1. Names and Suppliers of Biochemical Reagents and Kits

[0020]

[0021]

[0022] Experimental instruments and equipment

[0023] Table 2. Names and Suppliers of Experimental Instruments and Equipment

[0024]

[0025] laboratory animals

[0026] Male wild-type mice (C57BL / 6J) were purchased from Beijing Huafukang Biotechnology Co., Ltd. All animals were bred and housed in the SPF-grade environmental animal facility of the Beijing Institute of Cardiovascular and Pulmonary Diseases. Wild-type mice were males aged 10-12 weeks and weighing approximately 25-30g. All experimental procedures were performed according to the NIH's 1996 Guidelines for the Management and Use of Laboratory Animals and the experimental procedures stipulated by the Laboratory Animal Management Committee of Capital Medical University. All experimental animals were randomly assigned to groups.

[0027] Animal model testing

[0028] Pulse-type tailband blood pressure measurement

[0029] Using a pulse-type tail-cuff non-invasive blood pressure measurement system (Shanghai Aurcot Biotechnology Co., Ltd., China), mice were placed in a standardized animal facility. After calibration, the pulse-type tail-cuff system was activated, and the mice were placed in a dedicated measurement cage on a constant-temperature blanket (37℃) for 5–10 minutes until the mice stabilized. The tail pulse was then detected using a tail pulse receptor. Once the pulse stabilized, blood pressure measurement began. During the measurement, the temperature of the constant-temperature blanket and the number of measurements could be adjusted according to the mouse's condition, and the BP values ​​were recorded. Finally, the BP values ​​were read using the instrument (≥6 times), the highest and lowest values ​​were discarded, and the average value was taken as the final BP value for the mouse.

[0030] Intraperitoneal glucose tolerance test

[0031] Mice were fasted for 12 hours before undergoing a glucose tolerance test by intraperitoneal injection of glucose (2 g / kg dissolved in sterile water). Blood glucose levels (mmol / L) were measured using a glucometer via tail vein sampling before injection and at 15, 30, 60, 90, and 120 minutes after injection.

[0032] Small animal ultrasound

[0033] At weeks 0, 10, and 5 after drug administration, cardiac ultrasound data were acquired using a Vevo2100 high-resolution imaging system (Vevo2100; Visual Sonics) for small animals during HFpEF model establishment. The procedure was as follows: Mice were first anesthetized with 10% isoflurane gas, then placed on an ultrasound plate. Anesthesia was maintained using 5% isoflurane. Hair was removed from the mouse's chest using depilatory cream, and coupling gel was applied. Left ventricular outflow tract M-mode ultrasound was acquired using an ultrasound probe. LVEF and other systolic function parameters were obtained through short-axis M-mode scanning at the ventricular level. Diastolic function was measured at the mitral valve level using pulsed wave and tissue Doppler imaging. Apical four-chamber view of the anesthetized mice was obtained. Echocardiography was acquired under controlled body temperature. Isoflurane levels were reduced to 1.0-1.5% and adjusted to maintain a heart rate within the range of 400-500 beats / min. The following parameters were then calculated using analysis software: left ventricular end-diastolic diameter (LVDd), left ventricular end-systolic diameter (LVDs), left ventricular end-diastolic interventricular septal thickness (IVSd), left ventricular end-diastolic posterior wall thickness (LVPWd), left ventricular fractional shortening (FS), left ventricular ejection fraction (LVEF), peak diastolic velocity (E wave) through the mitral valve in early diastole, peak diastolic velocity (A wave) caused by atrial contraction in late diastole, isovolumetric relaxation time (IVRT), peak diastolic velocity of the myocardium in early diastole of the mitral annulus (E' wave), and early filling deceleration time (EDT). All parameters were measured at least three times, and the average values ​​were given.

[0034] Example 1: Construction of a heart failure model with preserved ejection fraction (HFpEF "two-hit" mouse model)

[0035] Male C57BL / 6J mice aged 8-12 weeks were randomly divided into two groups and placed in an animal room for feeding, maintaining a 12-hour light / dark cycle, with free access to food and water.

[0036] One group served as the control group (n=12): they were given a normal diet.

[0037] One group was the HFpEF group (n=24): they were given a high-fat diet (D12492) and L-NAME was dissolved in drinking water at a concentration of 0.5 g / L.

[0038] The phenotypic evaluation scheme for constructing the HFpEF model is as follows:

[0039] 1. The body weight of mice in both groups was measured at weeks 0 and 10 of feeding.

[0040] 2. At weeks 0 and 10 of feeding, small animal echocardiography was performed on mice in both groups to measure cardiac ejection fraction, degree of left ventricular diastolic dysfunction, degree of myocardial hypertrophy, and other cardiac function conditions.

[0041] 3. Blood pressure was measured in both groups of mice at weeks 0 and 10 of feeding using the pulse tail cuff method to measure blood pressure changes.

[0042] 4. At weeks 0 and 10 of feeding, mice in both groups underwent intraperitoneal glucose tolerance tests to measure their glucose tolerance.

[0043] During the 10th week of feeding, the following can be observed:

[0044] ①The HFpEF group had a significantly higher body weight than the control group, exhibiting an obese state;

[0045] ② In the HFpEF group, the left ventricular ejection fraction was ≥50%, indicating that ejection fraction was preserved; in the HFpEF group, E / e' was ≥40, indicating that there was left ventricular diastolic dysfunction.

[0046] ③In the HFpEF group, a systolic blood pressure ≥130 mmHg indicates hypertension;

[0047] ④ The HFpEF group showed glucose intolerance compared to the control group. This indicates that the model was successfully constructed.

[0048] Example 2: The therapeutic effect of CDK4 / 6 inhibitor (Palbociclib) on heart failure with preserved ejection fraction.

[0049] The cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor (Palbociclib) was ordered from Abmole Bioscience (China), catalog number M6167.

[0050] Using the HFpEF mouse model successfully constructed as in Example 1, the HFpEF group mice that had been continuously fed for 10 weeks were randomly divided into two groups.

[0051] (1) HFpEF group (n=12): Continue to feed high-fat diet (D12492), and dissolve L-NAME in drinking water at a concentration of 0.5g / L.

[0052] (2) Palbociclib treatment group (n=12): continued to feed a high-fat diet (D12492), dissolved L-NAME at a concentration of 0.5 g / L in drinking water, and at the same time, palbociclib was mixed into the high-fat diet and administered orally at a dose of 100 mg / kg / d.

[0053] The treatment effect was assessed after 5 weeks.

[0054] The drug administration observation indicators include:

[0055] 1. Five weeks after administration, small animal body composition analysis was performed on mice, and changes in total weight, lean body mass (fat-free body mass), and fat content were measured.

[0056] 2. Five weeks after administration, small animal echocardiography was performed on mice to measure cardiac function, including cardiac ejection fraction and the degree of left ventricular diastolic dysfunction.

[0057] 3. Five weeks after administration, blood pressure was measured in mice using the pulse tail cuff method to observe changes in blood pressure.

[0058] 4. Five weeks after administration, mice were subjected to an intraperitoneal glucose tolerance test to measure their glucose tolerance.

[0059] In the test indicators, total weight is divided into two components: fat and non-fat. The former is called fat mass (or obese body mass), and the latter is called lean mass (or fat-free body mass). This experiment measures the lean mass index.

[0060] Observation endpoint:

[0061] Five weeks after administration, all mouse hearts were collected, their weight was measured, and the length of the mouse tibia was measured. Lung tissue was collected and weighed to determine the degree of pulmonary congestion. White adipose tissue was collected for pathological staining (HE staining) to measure changes in adipocyte size.

[0062] The results showed that palbociclib treatment significantly improved the typical phenotype of HFpEF mice, specifically:

[0063] (1) The body weight of HFpEF mice was significantly reduced after 5 weeks of treatment with palbociclib. Figure 1 A);

[0064] (2) The ratio of heart weight to tibia length (HW / TL) was significantly reduced, approaching the normal value. Figure 1 B);

[0065] (3) The ratio of lean body mass (fat-free body mass) to fat content has significantly improved and returned to normal. Figure 1 C, D);

[0066] (4) Echocardiography results showed that the ejection fraction (LVEF) of mice in the Control group, HFpEF group, and palbociclib treatment group were all at normal levels. Figure 1 Compared with the HFpEF group, the palbociclib treatment group showed that diastolic function in mice returned to normal (E / e' significantly decreased), and the end-diastolic interventricular septal thickness (IVS, d) tended to decrease. Figure 1 F, G). It was also observed that systolic blood pressure (SBP) in mice returned to normal levels after palbociclib treatment. Figure 1 H);

[0067] (5) The lung weight ratio (wet weight / dry weight) shows a decreasing trend. Figure 1 I);

[0068] (6) Significant improvement in glucose tolerance ( Figure 1 J).

[0069] (7) HE staining of fat (under 200x microscope) showed that the size of adipocytes was significantly reduced after palbociclib treatment. Figure 3 AD, where A is the Control group, B is the HFpEF group, C is the palbociclib treatment group, and D is the quantitative result.

[0070] In summary, oral palbociclib can significantly improve the clinical symptoms of the HFpEF model.

[0071] Finally, it should be noted that the above embodiments are only used to help those skilled in the art understand the essence of the technical solution of the present invention, and are not intended to limit the scope of protection of the present invention.

Claims

1. Application of palbociclib, a cyclin-dependent kinase (CDK4 / 6) inhibitor, in the preparation of drugs for treating heart failure with preserved ejection fraction.

2. The application according to claim 1, characterized in that, The drugs mentioned are: drugs administered via the gastrointestinal tract, drugs administered via intravenous injection, and drugs administered via subcutaneous implantation.

3. The application according to claim 1 or 2, characterized in that, The drug contains a therapeutically effective amount of palbociclib and necessary pharmaceutical excipients.

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