Use of carnitine palmitoyltransferase 1 inhibitors in the treatment of heart failure with preserved ejection fraction

Abstract

The present invention relates to the use of carnitine palmitoyltransferase 1 inhibitors in the treatment of heart failure with preserved ejection fraction. The use in particular is the use of the carnitine palmitoyltransferase (CPT1) inhibitor Etomoxir for the prevention and / or treatment of heart failure with preserved ejection fraction by oral administration.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine, and in particular relates to an application of carnitine palmitoyltransferase 1 inhibitors in treating heart failure with preserved ejection fraction. BACKGROUND

[0002] Heart failure with preserved ejection fraction (HFpEF) refers to heart failure with left ventricular ejection fraction (LVEF) ≥ 50%, which is a multi-system disorder syndrome involving the heart, lungs, kidneys, skeletal muscles, adipose tissue, vascular system, immune and inflammatory signaling, and is characterized by hemodynamic disorders such as hypertension, myocardial hypertrophy and diastolic dysfunction. Among all heart failure (HF), HFpEF accounts for at least 50%. The 5-year all-cause mortality rate of HFpEF is > 50%, and the 1-year re-hospitalization rate is > 50%. Nearly 50% of HFpEF patients have 5 or more comorbidities, such as obesity, diabetes, pulmonary hypertension, etc., and thus have heterogeneity and complexity. So far, the pathophysiological mechanism of HFpEF is not fully understood, and thus it has always been a therapeutic difficulty in heart failure.

[0003] In large-scale randomized controlled trials of HFpEF, angiotensin-converting enzyme inhibitors (ACEI) or angiotensin receptor / brain peptide enzyme inhibitors (ARNI), beta receptor blockers, mineralocorticoid receptor antagonists (MRA), and other clinical drugs effective in treating heart failure with reduced ejection fraction (HFrEF) have not effectively reduced clinical endpoints. Trials conducted on other types of drugs (such as drugs targeting the nitric oxide-cyclic guanosine monophosphate pathway) have failed to improve clinical conditions or are neutral in terms of primary endpoints. The 2022 ESC Heart Failure Guidelines and the 2022 AHA / ACC / HFSA Guidelines suggest that HFpEF patients use diuretics and other symptomatic relief as needed, pay attention to the identification and treatment of comorbidities, and use sodium-glucose cotransporter 2 (SGLT2) inhibitors as a 2a-level recommendation. In the EMPEROR-Preserved, DELIVER, SOLOIST-WHF DELIVER three clinical trials, SGLT2 inhibitors effectively reduced the composite endpoint of cardiovascular death or heart failure hospitalization, becoming the only drug proven to improve the prognosis of HFpEF patients. However, in these clinical trials, only part of the heart failure patients with ejection fraction ≥ 50% were included, and as the LVEF increased, the drug's effect on reducing mortality weakened, indicating that SGLT2 inhibitors only benefit part of HFpEF patients.

[0004] So far, there is no specific treatment for reducing the morbidity and mortality of HFpEF patients, and it is necessary to explore other drugs for treating HFpEF.

[0005] Based on this, the present application is proposed. SUMMARY

[0006] The present application relates to the use of the carnitine palmitoyltransferase 1 (CPT1) inhibitor Etomoxir in the preparation of a medicament and / or a pharmaceutical composition for the treatment and / or prevention of heart failure with preserved ejection fraction.

[0007] Further, the medicament is a medicament for gastrointestinal administration, a medicament for intravenous injection, a medicament for subcutaneous embedding administration, preferably a medicament for gastrointestinal administration.

[0008] Further, the medicament and / or the pharmaceutical composition comprises a therapeutically effective amount of Etomoxir and necessary pharmaceutical excipients.

[0009] In a healthy heart, the main source of myocardial energy (about 60% to 80%) comes from fatty acid oxidation, and the rest comes from the oxidation of glucose or other substrates. Carnitine palmitoyltransferase 1 (CPT1) is a key enzyme in fatty acid oxidation, which is located on the outer side of the inner mitochondrial membrane, and its function is to transfer long-chain fatty acyl coenzyme A outside the mitochondria to the mitochondrial matrix for beta oxidation. Metabolic modulators are a new class of drugs that regulate cardiac metabolism without changing hemodynamics.

[0010] The present application has the beneficial effects of,

[0011] By using animal models to explore the pathological mechanism of HFpEF, a new drug, the carnitine palmitoyltransferase 1 (CPT1) inhibitor Etomoxir, which can treat HFpEF, is found.

[0012] In summary, the present application first reports the therapeutic effect of the carnitine palmitoyltransferase 1 (CPT1) inhibitor Etomoxir on heart failure with preserved ejection fraction. BRIEF DESCRIPTION OF DRAWINGS

[0013] Figure 1 , the therapeutic effect of CPT1 inhibitor (Etomoxir, Etomoxir) on the heart failure with preserved ejection fraction model of mice.

[0014] Figure 2 , the results of heart fat tissue pathological staining (HE staining) of the model mice before and after treatment. DETAILED DESCRIPTION

[0015] Biochemical reagents and kits

[0016] Table 1, biochemical reagents and kit names and suppliers

[0017]

[0018] Experimental instruments and equipment

[0019] Table 2, names of experimental instruments and equipment and suppliers

[0020]

[0021]

[0022] Experimental animals

[0023] Male wild-type mice (C57BL / 6J) were purchased from Beijing Huafukang Bioscience Co., Ltd. All animals were bred and raised in the SPF-level environment animal room of the Beijing Institute of Cardiovascular Disease. Wild mice should be male mice that are 10-12 weeks old and weigh about 25-30 g. All experimental operations were performed according to the NIH Guide for the Care and Use of Laboratory Animals established in 1996 and the experimental procedures specified by the Capital Medical University Experimental Animal Management Committee. All experimental animals were randomly grouped.

[0024] Animal model detection

[0025] Pulse tail-cuff method blood pressure measurement

[0026] The non-invasive blood pressure measurement system (Shanghai Alcor Bio Co., Ltd., China) was used. The mice were placed in a standardized animal room, the pulse tail-cuff pressure measurement system was turned on, and after calibration, the mice were placed in a special cage for pressure measurement on a constant temperature blanket (37°C) for 5-10 min. After the mouse's condition was stable, the mouse's tail pulse was sensed by the tail pulse sensor, and after the pulse was stable, the pressure measurement was started. During the measurement process, the temperature of the constant temperature blanket and the number of pressure measurements could be adjusted according to the mouse's condition, and the BP value was recorded. Finally, the BP value (≥6 times) was read by the instrument, the highest and lowest values were removed, and the average value was taken as the final BP value of the mouse.

[0027] Intraperitoneal glucose tolerance test

[0028] After the mice were fasted for 12 h, a glucose tolerance test was performed by intraperitoneal injection of glucose (2 g / kg dissolved in sterile water). Before injection and at 15, 30, 60, 90, and 120 minutes after injection, blood glucose levels (mmol / L) were measured by tail vein blood sampling and a blood glucose meter.

[0029] Small animal ultrasound

[0030] At the 0th, 10th week of HFpEF model construction and 5 weeks after drug administration, cardiac ultrasound data collection was performed using a small animal ultrasound instrument Vevo2100 high-resolution imaging instrument (Vevo2100; Visual Sonics). The process was as follows: first, the mice were anesthetized with isoflurane gas (10%), then placed on an ultrasonic metal plate, maintained in an anesthetized state using 5% isoflurane, the mouse chest hair was removed using depilatory cream, and the ultrasonic probe was used to collect the left ventricular outflow tract M-mode of the mouse heart. The LVEF and other systolic function indicators were obtained by scanning the short-axis M-mode mode of the ventricular level; the diastolic function was measured at the level of the mitral valve using pulse wave and tissue Doppler imaging, and the apical four-chamber view of the anesthetized mouse was obtained. The echocardiogram was collected under the condition of controlling the body temperature, and the isoflurane was reduced to 1.0-1.5%, and adjusted to keep the heart rate in the range of 400-500 times / min. Then the left ventricular end-diastolic diameter (LVDd), left ventricular end-systolic diameter (LVDs), interventricular septal thickness at end-diastole (IVSd), left ventricular posterior wall thickness at end-diastole (LVPWd), left ventricular fractional shortening (FS), left ventricular ejection fraction (LVEF), peak blood flow velocity through the mitral valve in early diastole (E wave), peak blood flow velocity caused by atrial contraction in late diastole (A wave), isovolumic relaxation time (IVRT), peak myocardial relaxation velocity in early diastole of the mitral annulus (E' wave), and early filling deceleration time (EDT) were calculated by analysis software. All parameters were measured at least 3 times, and the average value was given.

[0031] Example 1, Construction of a model of heart failure with preserved ejection fraction (HFpEF “two-hit” mouse model)

[0032] 8-12-week-old male C57BL / 6J mice were randomly divided into two groups and fed in an animal room, maintaining a 12-hour light / dark cycle, with free access to food and water.

[0033] One group was the control group (n=12): given a normal diet.

[0034] One group was the HFpEF group (n=24): given a high-fat diet (D12492), and L-NAME was dissolved in drinking water at a concentration of 0.5 g / L.

[0035] The phenotype evaluation scheme for the construction of the HFpEF model was as follows:

[0036] 1. At the 0th and 10th weeks of feeding, the body weight of the mice in both groups was measured.

[0037] 2. At the 0th and 10th weeks of feeding, small animal cardiac ultrasound was performed on the mice in both groups to measure the cardiac ejection fraction and the degree of left ventricular diastolic dysfunction.

[0038] 3. At the 0th and 10th weeks of feeding, the blood pressure of the two groups of mice was measured by pulse tail-cuff method to measure the changes in blood pressure.

[0039] 4. At the 0th and 10th weeks of feeding, the mice in the two groups were subjected to intraperitoneal glucose tolerance test to measure the glucose tolerance.

[0040] At the 10th week of feeding, it can be observed that:

[0041] ① The body weight of the HFpEF group increased significantly compared with the control group, showing obesity;

[0042] ② The left ventricular ejection fraction of the HFpEF group was ≥50%, i.e. there was ejection fraction preservation; the E / e' of the HFpEF group was ≥40, indicating left ventricular diastolic dysfunction;

[0043] ③ The systolic blood pressure of the HFpEF group was ≥130 mmHg, i.e. there was hypertension;

[0044] ④ The HFpEF group showed glucose intolerance compared with the control group. The above indicates that the model is successfully constructed.

[0045] Example 2, Therapeutic effect of CPT1 inhibitor (Etomoxir) on ejection fraction preserved heart failure

[0046] Carnitine palmitoyltransferase (CPT1) inhibitor Etomoxir (Ethomoxir) was ordered from Abmole Bioscience (China) with the item number M9048.

[0047] Sodium-glucose cotransporter 2 (SGLT2) inhibitor Empagliflozin (Empagliflozin) was ordered from Abmole Bioscience (China) with the item number M2667.

[0048] Using the HFpEF mouse model successfully constructed as in Example 1, the HFpEF group of mice fed for 10 weeks above were randomly divided into two groups,

[0049] (1) HFpEF group (n = 8): continue to feed high-fat diet (D12492), and dissolve L-NAME in drinking water at a concentration of 0.5 g / L.

[0050] (2) Ethomoxir treatment group (n = 8): continue to feed high-fat diet (D12492), dissolve L-NAME in drinking water at a concentration of 0.5 g / L, and orally administer ethomoxir at a dose of 20 mg / kg / d by mixing it in the high-fat diet feed.

[0051] (3) Empagliflozin treatment group (n = 8): continue to feed high-fat diet (D12492), dissolve L-NAME in drinking water at a concentration of 0.5 g / L, and orally administer Empagliflozin at a dose of 10 mg / kg / d by mixing it in the high-fat diet feed. This group serves as a positive control.

[0052] The treatment effect is detected after 5 weeks.

[0053] The observation indicators of administration include:

[0054] 1. After 5 weeks of administration, small animal body composition analysis is performed on the mice to measure changes in total body weight, lean body weight (lean body weight), and fat content.

[0055] 2. After 5 weeks of administration, small animal echocardiography is performed on the mice to measure cardiac ejection fraction, left ventricular diastolic dysfunction, and other cardiac function.

[0056] 3. After 5 weeks of administration, pulse tail cuff blood pressure measurement is performed on the mice to measure changes in blood pressure.

[0057] 4. After 5 weeks of administration, intraperitoneal glucose tolerance test is performed on the mice to measure glucose tolerance.

[0058] In the detection indicators, total body weight is divided into fat and non-fat components. The former is called fat weight (or fat weight), and the latter is called lean body weight (or lean body weight). In this experiment, the lean body weight indicator is detected.

[0059] Observation endpoints:

[0060] After 5 weeks of administration, the hearts of all mice are collected, the heart weight is measured, the tibia length of the mice is measured, the lung tissue is collected to measure the dry and wet weights to detect the degree of pulmonary congestion, and the white adipose tissue is collected for pathological staining (HE staining) to measure the size changes of adipocytes.

[0061] The results show that CPT1 inhibitor Etomoxir significantly improves the phenotype of HFpEF mice after treatment, and Etomoxir is superior to Empagliflozin in treating HFpEF mice, and Etomoxir is superior to Empagliflozin in improving the thickness of the interventricular septum, the degree of pulmonary congestion, and the size of adipocytes. Specifically:

[0062] (1) The body weight of HFpEF mice treated with Etomoxir and Empagliflozin for 5 weeks decreased significantly ( Figure 1 A);

[0063] (2) The ratio of heart weight to tibia length (HW / TL) decreased significantly, close to the normal value ( Figure 1 B);

[0064] (3) The ratio of lean body mass (lean body mass) and fat content was significantly improved, and returned to normal( Figure 1 C, D);

[0065] (4) The results of cardiac ultrasound showed that the left ventricular ejection fraction (LVEF) of the mice in the Control group, the HFpEF group, the ethmoxifer treatment group and the empagliflozin treatment group were all at a normal level( Figure 1 E), compared with the HFpEF group, the diastolic function of the mice in the ethmoxifer treatment group and the empagliflozin treatment group returned to normal (E / e' was significantly reduced) Figure 1 F), End diastolic chamber volume after empagliflozin treatment The interventricular septum thickness (IVS, d) has a decreasing trend, while it is significantly reduced after ethmozine treatment Figure 1 G) The systolic blood pressure (SBP) of the mice in the ethmoxifer treatment group and the empagliflozin treatment group returned to normal level( Figure 1 H);

[0066] (5) Glucose tolerance disorder was significantly improved( Figure 1 J).

[0067] (6) Ethmoxane significantly reduced the lung weight ratio (wet / dry) after treatment Figure 1 I), lung congestion was significantly alleviated While empagliflozin treatment was associated with a significant improvement in LV diastolic function, no significant improvement was observed after treatment with placebo .

[0068] (7) Fat HE staining (200x) showed that the size of adipocytes was reduced after empagliflozin treatment, while the size of adipocytes returned to normal after ethmoxifer treatment( Figure 2 A-E, where A is the Control group, B is the HFpEF group, C is the ethmoxifer treatment group, D is the empagliflozin treatment group, and E is the quantitative result).

[0069] In summary, oral administration of ethmoxifer can significantly improve the clinical symptoms of the HFpEF model, and the therapeutic effect is significantly better than that of the SGLT2 inhibitor empagliflozin.

[0070] Finally, it should be noted that the above examples are only used to help those skilled in the art to understand the essence of the technical solutions of the present application, and are not used to limit the protection scope of the present application.

Claims

1. Use of Etomoxir in the manufacture of a medicament for the treatment of heart failure with preserved ejection fraction.

2. Use according to claim 1, characterized in that, The medicament is a medicament for administration via the gastrointestinal tract.

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