Application of fatty acid-binding protein 4 inhibitors in the treatment of heart failure with preserved ejection fraction

Abstract

This invention relates to the application of fatty acid-binding protein 4 (FABP4) inhibitors in the treatment of heart failure with preserved ejection fraction. Specifically, the application involves the oral administration of the fatty acid-binding protein 4 (FABP4) inhibitor BMS309403 for the prevention and / or treatment of heart failure with preserved ejection fraction.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of fatty acid-binding protein 4 inhibitors in the treatment of heart failure with preserved ejection fraction. Background Technology

[0002] Heart failure with preserved ejection fraction (HFpEF) is defined as heart failure with a left ventricular ejection fraction (LVEF) ≥50%, characterized by diastolic dysfunction and preservation of ejection fraction. HFpEF accounts for half of all heart failure cases and is associated with a variety of comorbidities, including diabetes, hypertension, and restrictive cardiomyopathy. In HFpEF, chronic systemic inflammation and metabolic disturbances affect not only the myocardium but also other organs such as the kidneys, lungs, and skeletal muscles. HFpEF accounts for at least 50% of all heart failure (HF) cases. The 5-year survival rate for HFpEF is only 35%. The pathophysiological mechanisms of HFpEF are complex, making it a persistent challenge in the treatment of heart failure.

[0003] To date, the pathophysiological mechanisms of HFpEF are not fully understood, and there is no specific treatment to reduce the morbidity and mortality of HFpEF patients. It is essential to explore other drugs for the treatment of HFpEF.

[0004] Currently, commonly used drugs for the treatment of HFpEF mainly include beta-blockers, angiotensin-converting enzyme inhibitors (ACEIs) or angiotensin receptor / neprilysin inhibitors (ARNIs), and mineralocorticoid receptor antagonists (MRAs), which are effective in treating heart failure with reduced ejection fraction (HFrEF).

[0005] (1) Beta-blockers can effectively reduce the heart rate of patients. However, in a study of patients with diastolic dysfunction, the use of beta-blockers resulted in high BNP levels and unexpected worsening of heart failure symptoms. Although reducing heart rate is beneficial for preventing excessive ventricular rate in patients with atrial fibrillation, preventing tachyarrhythmias, and treating angina symptoms, its long-term efficacy is uncertain.

[0006] (2) ACEIs can reduce the production of angiotensin II, inhibit angiotensin-converting enzyme, thereby reducing the production of angiotensin II and aldosterone secretion, thus alleviating water and sodium retention. ARNIs have a dual effect of angiotensin receptor blocking and natriuretic peptide system activation, and have the effects of sodium excretion, diuresis to reduce volume overload and anti-myocardial fibrosis. However, in its clinical studies on the treatment of HFpEF, ACEI / ARNI did not significantly reduce the composite endpoint of death and total hospitalization for heart failure in HFPEF patients.

[0007] (3) MRA drugs have a clear effect in HFrEF, but the effect is poor in HFpEF, possibly because the pathophysiological effect of RAAS is not significant as LVEF increases.

[0008] The 2022 ESC Heart Failure Guidelines and the 2022 AHA / ACC / HFSA Guidelines recommend that patients with heart failure with paroxysmal ejection fraction (HFpEF) use diuretics and other medications to relieve symptoms as needed, and classify sodium-glucose cotransporter 2 inhibitors (SGLT2i) as a level 2a recommendation. In the EMPEROR-PRESERVED trial, the SGLT2i empagliflozin reduced the risk of composite CV death or total HF hospitalization in HF patients with LVEF > 40%. In the SOLOIST-WHF trial (Effect of Sopagliflozin on Heart Failure Worsening in Patients with Type 2 Diabetes), the dual SGLT2 and SGLT1 inhibitor sopagliflozin reduced the primary endpoints of CV death and HF hospitalization in diabetic patients and HF worsening (HFrEF and HFpEF). The dapagliflozin DELIVER study further elucidates the benefit of SGLT2i for patients with HFpEF. However, in these clinical trials, only a subset of patients had an ejection fraction ≥ 50%, and the mortality-reducing effect of the drugs diminished with increasing LVEF, indicating that SGLT2i are only effective in a subset of HFpEF patients. Therefore, it is crucial to explore safer and more effective drugs that can alleviate HFpEF.

[0009] Based on this, the present invention is proposed. Summary of the Invention

[0010] This invention primarily relates to the use of fatty acid-binding protein 4 (FABP4) inhibitor BMS309403 in the preparation of medicaments and / or pharmaceutical compositions for the treatment and / or prevention of heart failure with preserved ejection fraction.

[0011] 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.

[0012] Furthermore, the drug and / or pharmaceutical composition contains a therapeutically effective amount of BMS309403, as well as necessary pharmaceutical excipients.

[0013] The beneficial effects of this invention are as follows:

[0014] Fatty acid-binding protein (FABP) family members are most abundantly expressed in adipocytes, with fatty acid-binding protein 4 (FABP4) being the most prominent. FABP4 is a lipid-binding protein that participates not only in lipid transport but also regulates glucose and lipid homeostasis. Clinical studies have shown elevated FABP4 expression in the serum of patients with heart failure. BMS309403, also known as a selective FABP4 inhibitor, competitively inhibits the binding of endogenous free fatty acids to fatty acid-binding vesicles of FABP4.

[0015] In this invention, by using an animal model to explore the pathological mechanism of HFpEF, it was found that the fatty acid binding protein 4 (FABP4) inhibitor BMS309403 can significantly alleviate the related symptoms of HFpEF. Attached Figure Description

[0016] Figure 1 The therapeutic effect of FABP4 inhibitor (BMS309403) on a mouse model of heart failure with preserved ejection fraction.

[0017] Figure 1 A. After treatment with BMS309403, the body weight of HFpEF mice decreased significantly;

[0018] Figure 1 B. After treatment with BMS309403, the ratio of heart weight to tibia length (HW / TL) in HFpEF mice was significantly reduced to near normal values.

[0019] Figure 1 C. After treatment with BMS309403, the lean body mass (fat-free body mass) of HFpEF mice was significantly improved;

[0020] Figure 1 D. After treatment with BMS309403, the proportion of fat content in HFpEF mice was significantly improved;

[0021] Figure 1 E. Echocardiography results showed that the ejection fraction (LVEF) of mice in the Control group, HFpEF group, and BMS309403 treatment group were all at normal levels.

[0022] Figure 1 F. Compared with the HFpEF group, the diastolic function of mice in the BMS309403 treatment group was significantly restored (E / e' was significantly reduced);

[0023] Figure 1 After treatment with G and BMS309403, systolic blood pressure (SBP) in HFpEF mice decreased significantly.

[0024] Figure 1After treatment with H and BMS309403, glucose intolerance in HFpEF mice was significantly improved.

[0025] Figure 2 Structural formula of compound BMS309403.

[0026] Figure 3 1. Pathological staining results of cardiac adipose tissue (HE staining) in model mice before and after treatment.

[0027] Figure 3 A. Control group;

[0028] Figure 3 Group B, HFpEF;

[0029] Figure 3 C, BMS309403 treatment group;

[0030] Figure 3 D. Quantitative results. Detailed Implementation

[0031] Biochemical reagents and kits

[0032] Table 1. Names and Suppliers of Biochemical Reagents and Kits

[0033]

[0034]

[0035] Experimental instruments and equipment

[0036] Table 2. Names and Suppliers of Experimental Instruments and Equipment

[0037]

[0038] laboratory animals

[0039] 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 Department of Capital Medical University. All experimental animals were randomly assigned to groups.

[0040] Animal phenotyping

[0041] Pulse-type tailband blood pressure measurement

[0042] 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 turned on, 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 sensed using a tail pulse receptor, and blood pressure measurement began after the pulse stabilized. During the measurement, the temperature of the constant-temperature blanket and the number of blood pressure measurements could be adjusted according to the mouse's condition, and the BP values ​​were recorded. Finally, the BP values ​​were read by 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.

[0043] Intraperitoneal glucose tolerance test

[0044] 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.

[0045] Small animal ultrasound

[0046] 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.

[0047] Example 1: Construction of a heart failure model with preserved ejection fraction (HFpEF "two-hit" mouse model)

[0048] 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.

[0049] One group served as the control group (n=12): they were given a normal diet.

[0050] 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.

[0051] The phenotypic evaluation scheme for constructing the HFpEF model is as follows:

[0052] 1. The body weight of mice in both groups was measured at weeks 0 and 10 of feeding.

[0053] 2. At weeks 0 and 10 of feeding, small animal echocardiography was performed on mice in both groups to measure cardiac function, such as cardiac ejection fraction and the degree of left ventricular diastolic dysfunction.

[0054] 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.

[0055] 4. At weeks 0 and 10 of feeding, mice in both groups underwent intraperitoneal glucose tolerance tests to measure their glucose tolerance.

[0056] During the 10th week of feeding, the following can be observed:

[0057] ①The HFpEF group had a significantly higher body weight than the control group, exhibiting an obese state;

[0058] 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.

[0059] ③ The HFpEF group had a systolic blood pressure ≥130 mmHg, indicating hypertension; ④ The HFpEF group showed glucose intolerance compared to the control group. These findings indicate that the model was successfully constructed.

[0060] Example 2: The therapeutic effect of administering a FABP4 inhibitor (BMS309403) on heart failure with preserved ejection fraction.

[0061] The fatty acid-binding protein (FABP4) inhibitor BMS309403 was ordered from Abmole Bioscience (China), catalog number M6265.

[0062] 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.

[0063] (1) HFpEF group (n=8): Continue to feed high-fat diet (D12492), and dissolve L-NAME in drinking water at a concentration of 0.5g / L.

[0064] (2) BMS309403 treatment group (n=8): Continue to feed a high-fat diet (D12492), dissolve L-NAME in drinking water at a concentration of 0.5g / L, and at the same time, mix BMS309403 into the high-fat diet and administer orally at a dose of 15mg / kg / d.

[0065] The treatment effect was assessed after 5 weeks.

[0066] The drug administration observation indicators include:

[0067] 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.

[0068] 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.

[0069] 3. Five weeks after administration, blood pressure was measured in mice using the pulse tail cuff method to observe changes in blood pressure.

[0070] 4. Five weeks after administration, mice were subjected to an intraperitoneal glucose tolerance test to measure their glucose tolerance.

[0071] Observation endpoint:

[0072] Five weeks after administration, all mouse hearts were collected, their weight was measured, and the length of the mouse tibia was measured. White adipose tissue was collected for pathological staining (HE staining) to measure changes in adipocyte size.

[0073] 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.

[0074] The results showed that the typical phenotype of HFpEF mice was significantly improved after BMS309403 treatment, specifically:

[0075] (1) The body weight of HFpEF mice treated with BMS309403 for 5 weeks was significantly reduced. Figure 1 A);

[0076] (2) The ratio of heart weight to tibial length (HW / TL) was significantly reduced to near normal. Figure 1 B);

[0077] (3) The ratio of lean body mass (fat-free body mass) to fat content was significantly improved. Figure 1 C, D);

[0078] (4) Echocardiography results showed that the ejection fraction (LVEF) of mice in the Control group, HFpEF group, and BMS309403 treatment group were all at normal levels. Figure 1 Compared with the HFpEF group, the diastolic function of mice in the BMS309403 treatment group was significantly restored (E / e' was significantly reduced), approaching normal levels. Figure 1 F), and it was also observed that BMS309403 treatment significantly reduced systolic blood pressure (SBP) in mice. Figure 1 G);

[0079] (5) Glucose intolerance significantly improved and returned to normal. Figure 1 H).

[0080] (6) HE staining of fat (under 200x microscope) showed that the size of adipocytes was significantly reduced after BMS309403 treatment. Figure 3 AD, where A is the Control group, B is the HFpEF group, C is the BMS309403 treatment group, and D is the quantitative result.

[0081] In summary, oral administration of BMS309403 can significantly improve the clinical symptoms of the HFpEF model.

[0082] 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. The use of fatty acid-binding protein 4 (FABP4) inhibitor BMS309403 in the preparation of medicaments and / or pharmaceutical compositions for the treatment of heart failure with preserved ejection fraction; wherein the medicament is administered via the gastrointestinal tract.

2. The application according to claim 1, characterized in that, The drug and / or pharmaceutical composition contains a therapeutically effective amount of BMS309403 and necessary pharmaceutical excipients.

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