Effects induced by sodium-glucose cotransporter inhibitors (SGLTi) in subjects suffering from cardiovascular disease

Abstract

The present invention relates to a method of detecting effects induced by a sodium-glucose cotransporter inhibitor (SGLTi) in a subject suffering from a cardiovascular disease, the method comprising a) determining the amount of fatty acid binding protein 3 (FABP3) in a sample of the subject treated with SGLTi; and b) comparing the amount of FABP3 to a reference amount. Computer-implemented methods, computer program products, devices and kits for performing the methods of the invention are also encompassed. The invention further relates to the use of FABP3, the use of FABP3 and a marker of cardiac injury, and / or the use of a detection agent of FABP3 or the use of a detection agent of FABP3 and a marker of cardiac injury for detecting effects induced by a sodium-glucose cotransporter inhibitor (SGLTi) in a subject suffering from cardiovascular disease.

Description

Technical Field

[0001] The present invention relates to a method for detecting an effect induced by a sodium-glucose cotransporter inhibitor (SGLTi) in a subject with cardiovascular disease, the method comprising a) determining the amount of fatty acid binding protein 3 (FABP3) in a sample from a subject treated with SGLTi; and b) comparing the amount of FABP3 to a reference amount. Also encompassed are computer-implemented methods, computer program products, apparatus, and kits for performing the methods of the invention. The present invention further relates to (i) the use of FABP3 or FABP3 and a marker of cardiac damage, and (ii) the use of FABP3 or a detection agent for FABP3 and a marker of cardiac damage, for detecting an effect induced by a sodium-glucose cotransporter inhibitor (SGLTi) in a subject with cardiovascular disease. Background Art

[0002] Cardiovascular disease seriously affects cardiac function and usually leads to the progression of heart failure, which is the main cause of morbidity and mortality in the Western Hemisphere. Heart failure (HF) is a complex clinical syndrome that can be caused by any structural or functional heart disease, which damages the ability of ventricular filling or ejection and the ability to protect the body's metabolic demand for blood / oxygen supply. HF affects approximately 64 million patients worldwide, and due to the increasing burden of aging population, HF comorbidities and risk factors and the extension of survival after myocardial infarction, its prevalence is rising (see, e.g., Castiglione V, Aimo A, Vergaro G, Saccaro L, Passino C and Emdin M. Biomarkers for the diagnosis and management of heart failure. Heart Failure Reviews (2022) 27: 625-643).

[0003] The goals of heart failure treatment are to improve survival, prevent hospitalization, and relieve symptoms and signs of heart failure (HF), such as congestion, dyspnea, and fatigue (McMurray JJ, Adamopoulos S, Anker SD, Auricchio A, Bohm M, Dickstein K, et al. ESC guidelines for the diagnosis and treatment of acute and chronic heart failure 2012: The Task Force for the Diagnosis and Treatment of Acute and Chronic Heart Failure 2012 of the European Society of Cardiology. Developed in collaboration with the Heart Failure Association (HFA) of the ESC. European journal of heart failure. 2012; 14(8):803-69). Current standard treatments for patients with heart failure include beta-blockers, RAAS inhibitors, angiotensin receptor-neprilysin inhibitors, and diuretics to suppress neurohormones, reduce volume overload, and thereby improve cardiac contractility (Ponikowski P, Voors AA, Anker SD, Bueno H, Cleland JGF, Coats AJS et al. 2016 ESC Guidelines for the diagnosis and treatment of acute and chronic heart failure: The Task Force for the diagnosis and treatment of acute and chronic heart failure of the European Society of Cardiology (ESC) Developed with the special contribution of the Heart Failure Association (HFA) of the ESC. European heart journal. 2016; 37(27): 2129-200). Despite the important cardiovascular benefits, there is still a risk of acute decompensation, hospitalization, and death, and further treatment options are needed.Thus, although available treatment options can reduce morbidity and mortality in patients with HF, the relative number of eligible patients receiving these treatments remains unsatisfactorily low (O'Donoghue M. & Braunwald E., Nat. Rev. Cardiol. 2010;7:13-20). Therefore, a goal of modern medicine is to provide personalized or individualized treatment guidance that takes into account the individual needs or risks of the patient. However, particularly for recently available treatment options, the mode of action may not yet be fully understood, and therefore a better understanding of such treatment options is first needed.

[0004] Sodium-glucose cotransporter (SGLT) inhibitors, primarily inhibitors such as sodium-glucose cotransporter 2 inhibitors (SGLT2i) and sodium-glucose cotransporter 1 inhibitors (SGLT1i), have attracted attention as therapeutic targets. Sodium-glucose cotransporters (SGLTs) constitute a large family of membrane proteins with at least six different isoforms in humans, and mediate glucose transport across the cell membrane, particularly across the apical membrane on the lumen side (see, for example, Sabino-Silva R et al., The Na(+) / glucose cotransporters: from genes to therapy. Brazilian Journal of Medical and Biological Research 2010. 43(11): 1019–26). SGLT inhibitors have been developed as antidiabetic treatments by inhibiting glucose reabsorption and increasing glucose excretion into the urine (see, e.g., Ferrannini E. Sodium-glucose co-transporters and their inhibition: clinical physiology. Cell Metab 2017; 26: 27–38). In addition, SGLT2i have been found to protect against cardiovascular outcomes, leading to changes in clinical recommendations and practices (see, e.g., Professional Practice Committee. Standards of medical care in diabetes-2019. Diabetes Care 2019; 42 (Suppl 1): S3). The mechanism of action of SGLTi in patients with cardiovascular disease is not fully understood. Therefore, there is a need to better understand SGLTi treatment in patients with cardiovascular disease. Markers that allow assessment / visualization / detection of the effects induced by sodium-glucose co-transporter inhibitors (SGLTi) in subjects with cardiovascular disease can also help to enhance such understanding.

[0005] The technical problem underlying the present invention can therefore be seen as providing means and methods for meeting the aforementioned needs. This technical problem is solved by the claims and the embodiments characterized in the following. Summary of the Invention

[0006] The present invention provides a method for detecting an effect induced by a sodium-glucose cotransporter inhibitor (SGLTi) in a subject suffering from cardiovascular disease, the method comprising a) determining the amount of fatty acid binding protein 3 (FABP3) in a sample from a subject treated with SGLTi; and b) comparing the amount of FABP3 to a reference amount. Furthermore, the present invention provides a computer-implemented method for performing the method of the present invention, the computer-implemented method comprising: a) means for determining the amount of FABP3 or the amount of FABP3 and a cardiac injury marker in a sample from a subject treated with SGLTi, wherein the subject suffers from cardiovascular disease; and b) means for comparing the amount of FABP3 to a reference amount, comparing the amount of FABP3 and the amount of the cardiac injury marker to a reference amount, or generating a ratio between the amount of FABP3 and the amount of the cardiac injury marker and comparing the ratio to a reference ratio.

[0007] The present invention also provides a computer program product comprising program code for executing the computer-implemented method of the present invention when run on at least one computer.

[0008] The present invention also provides a device for performing the method of the present invention, comprising: a) means for determining the amount of FABP3 or the amount of FABP3 and a cardiac damage marker in a sample of a subject treated with SGLTi, wherein the subject suffers from cardiovascular disease; and b) means for comparing the amount of FABP3 with a reference amount, comparing the amount of FABP3 and the amount of the cardiac damage marker with a reference amount, or generating a ratio between the amount of FABP3 and the amount of the cardiac damage marker and comparing the ratio with a reference ratio.

[0009] The present invention also provides a kit for performing the method of the present invention, comprising: a) a means for determining the amount of FABP3 or the amount of FABP3 and a cardiac marker in a sample of a subject treated with SGLTi, wherein the subject suffers from cardiovascular disease; and b) a means for comparing the amount of FABP3 with a reference amount, comparing the amount of FABP3 and the amount of the cardiac marker with a reference amount, or generating a ratio between the amount of FABP3 and the amount of the cardiac marker and comparing the ratio with a reference ratio.

[0010] The present invention also provides (i) uses of FABP3, uses of FABP3 and cardiac damage markers, and (ii) uses of detection agents for FABP3 or FABP3 and cardiac damage markers for detecting effects induced by sodium-glucose cotransporter inhibitors (SGLTi) in subjects with cardiovascular disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The invention will be better understood with reference to the detailed description when considered in conjunction with the non-limiting examples and accompanying drawings, in which:

[0012] Figure 1 The initiation of SGLT2i therapy was associated with an increase in FABP3 plasma concentrations. FABP3 concentrations are expressed in ng / mL; boxplots show median concentrations, lower quartile boundaries, and upper quartile boundaries. V0: baseline (median FABP3 = 37.82), V1: 4 weeks after initiation of SGLT2i therapy (median FABP3 = 40.86), V2: 3 months (median FABP3 = 45.14), V3: 6 months (median FABP3 = 44.36), and V4: 12 months (FABP3 = 44.47).

[0013] Figure 2 Figure : Changes in FABP3 concentrations over time after initiation of SGLT2i therapy in patients with high baseline FABP3 concentrations (above the median concentration of 37.8 ng / mL). FABP3 concentrations are expressed in ng / mL; boxplots show the median concentration, lower quartile, and upper quartile boundaries. V0: Baseline (median FABP3 = 47.97), V1: Week 4 after initiation of SGLT2i therapy (median FABP3 = 53.56), V2: 3 months (median FABP3 = 54.73), V3: 6 months (median FABP3 = 54.51), V4: 12 months (FABP3 = 53.23).

[0014] Figure 3 Figure : Changes in FABP3 concentrations over time after initiation of SGLT2i therapy in patients with low baseline FABP3 concentrations (below the median concentration of 37.8 ng / mL). FABP3 concentrations are expressed in ng / mL; boxplots show the median concentration, lower quartile, and upper quartile boundaries. V0: Baseline (median FABP3 = 29.05), V1: Week 4 after initiation of SGLT2i therapy (median FABP3 = 31.41), V2: 3 months (median FABP3 = 38.86), V3: 6 months (median FABP3 = 36.41), V4: 12 months (FABP3 = 36.76).

[0015] Figure 4 Figure : Changes in the FABP3 / NT-proBNP ratio over time after initiation of SGLT2i therapy. FABP3 concentrations are expressed in ng / mL and NT-proBNP concentrations in pg / mL; boxplots show the median FABP3 / NT-proBNP ratio, lower quartile boundaries, and upper quartile boundaries. V0: Baseline (median FABP3 / NT-proBNP ratio = 0.026), V1: Week 4 after initiation of SGLT2i therapy (median FABP3 / NT-proBNP ratio = 0.032), V2: Month 3 (median FABP3 / NT-proBNP ratio = 0.039), V3: Month 6 (median FABP3 / NT-proBNP ratio = 0.046), and V4: Month 12 (median FABP3 / NT-proBNP ratio = 0.033).

[0016] Figure 5 Figure : Changes in circulating NT-proBNP concentrations over time after initiation of SGLT2i therapy. NT-proBNP concentrations are expressed in pg / mL; boxplots show median concentrations, lower quartile boundaries, and upper quartile boundaries. V0: baseline (median NT-proBNP = 1396.30), V1: 4 weeks after initiation of SGLT2i therapy (median NT-proBNP = 1115.68), V2: 3 months (median NT-proBNP = 1191.95), V3: 6 months (median NT-proBNP = 889.21), and V4: 12 months (median NT-proBNP = 1172.84).

[0017] Figure 6 Figure : Changes in the circulating FABP3 / GDF-15 ratio over time after initiation of SGLT2i therapy. FABP3 concentrations are expressed in ng / mL and GDF-15 concentrations in pg / mL; boxplots show the median FABP3 / GDF-15 ratio, lower quartile boundaries, and upper quartile boundaries. V0: Baseline (median FABP3 / GDF-15 ratio = 0.017), V1: Week 4 after initiation of SGLT2i therapy (median FABP3 / GDF-15 ratio = 0.016), V2: Month 3 (median FABP3 / GDF-15 ratio = 0.016), V3: Month 6 (median FABP3 / GDF-15 ratio = 0.016), and V4: Month 12 (median FABP3 / GDF-15 ratio = 0.019).

[0018] Figure 7Figure : Changes in circulating GDF-15 concentrations over time after initiation of SGLT2i therapy. GDF-15 concentrations are expressed in pg / mL; boxplots show median concentrations, lower and upper quartile boundaries. V0: baseline (median GDF-15 = 2439.34), V1: 4 weeks after initiation of SGLT2i therapy (median GDF-15 = 2657.16), V2: 3 months (median GDF-15 = 3036.18), V3: 6 months (median GDF-15 = 2414.01), V4: 12 months (median GDF-15 = 2039.85). DETAILED DESCRIPTION

[0019] The present invention relates to a method for detecting an effect induced by a sodium-glucose co-transporter inhibitor (SGLTi) in a subject suffering from a cardiovascular disease, the method comprising (a) determining the amount of fatty acid binding protein 3 (FABP3) in a sample of a subject treated with SGLTi; (b) comparing said amount of FABP3 with a reference amount. Thus, the present invention is based on the finding that the effect induced by SGLTi in a subject suffering from a cardiovascular disease is associated with the amount of FABP3. In particular, as demonstrated in the experimental part herein, large-scale analysis of samples showed that in patients suffering from heart failure, SGLT2i treatment was associated with the amount of FABP3 in said patients; see Figure 1 . FABP3 can therefore be used to detect / visualize / assess the effects induced by SGLTi in subjects with cardiovascular disease. In particular, as demonstrated in the experimental part of this article, the concentration or amount of FABP3 increases with treatment with SGLTi (and thereby visualizes or assesses SGLTi treatment), wherein the increase is evident for at least up to 12 months. Therefore, the effects induced by SGLTi can be reliably detected (visualized). "Detection" can therefore be regarded as "visualization" or "assessment". In addition, the method of the present invention allows monitoring the effects induced by SGLTi in subjects with cardiovascular disease by: (a) determining the amount of fatty acid binding protein 3 (FABP3) in a sample of a subject treated with SGLTi; (b) comparing the amount of FABP3 with a reference amount. Monitoring and / or detecting (such as visualizing or assessing) the effects induced by SGLTi based on the amount of FABP3 can, for example, be used to develop personalized treatment plans, such as for guiding, adjusting or changing SGLTi therapy in patients with cardiovascular disease, and can therefore support doctors in making clinical decisions about SGLTi therapy.

[0020] Sodium-glucose cotransporter (SGLT) is located in the renal proximal tubular epithelium and functions mainly by reabsorb- ing filtered glucose (see, e.g., Bakris GL, Fonseca VA, Sharma K, Wright EM. Renalsodium-glucose transport:role in diabetes mellitus and potential clinical implications. Kidney international. 2009; 75(12): 1272-7; Ferrannini E, Murthy AC, Lee Y, Muscelli E, Weiss S, Ostroff RM, Sattar N, Williams SA and Ganz P. Mechanisms of Sodium-Glucose Cotransporter 2 Inhibition: Insights From Large-Scale Proteomics. Diabetes Care (2020); 43: 2183-2189). It is reported that SGLT inhibitors can lead to urinary excretion of glucose, sodium and water, thereby leading to hemodynamic changes and glycemic improvement in patients with type 2 diabetes. In addition, it is reported that SGLT2i can reduce the risk of cardiovascular events. Although there is a diuretic effect, it is suggested that SGLT inhibitors may also have further cardioprotective local and systemic mechanisms of action that are not yet clear. It is not desirable to be bound by theory, an explanation of the beneficial therapeutic effects of SGLT2i in cardiovascular diseases (such as heart failure) may be due to the main effect of SGLT2i on proximal renal tubules in several body function domains Inducing a series of physiological consequences (see, for example, Ferrannini E, Murthy AC, Lee Y, Muscelli E, Weiss S, Ostroff RM, Sattar N, Williams SA and Ganz P.Mechanisms of Sodium-Glucose Cotransporter 2 Inhibition:Insights From Large-Scale Proteomics.Diabetes Care (2020); 43:2183-2189).In addition, it has been reported that SGLT2i may increase natriuresis and osmotic diuresis, accompanied by volume contraction and increased hematocrit, which may be responsible for the reduction in blood pressure and arterial stiffness, which can improve cardiac function by reducing preload and postload (see, for example, Verma S, McMurray JJV. SGLT2 inhibitors and mechanisms of cardiovascular benefit: a state of-the-art review. Diabetologia 2018; 61: 2108–2117). In summary of this review, the mechanism by which SGLT2i exert its effects in subjects with cardiovascular disease is not yet fully understood. It has been reported that SGLTi can induce a fasting-like metabolic paradigm involving a metabolic switch from carbohydrate to lipid utilization and ketogenesis, which activates nutrient deprivation pathways and helps maintain energy homeostasis and cardiorenal protection (see, e.g., Gao YM, Feng ST, Wen Y, Tang T, Wang B, Liu BC. Cardiorenal protection of SGLT2 inhibitors-Perspectives from metabolic reprogramming. EBioMedicine. 2022; 83: 104215. doi: 10.1016 / j.ebiom.2022.104215), such as evidence of changes in FABP3 levels in diabetic non-HF patients (Ferrannini E, Murthy AC, Lee Y, Muscelli E, Weiss S, Ostroff RM, Sattar N, Williams SA and Ganz P. Mechanisms of Sodium-Glucose Cotransporter 2 Inhibition: Insights From Large-Scale Proteomics. Diabetes Care (2020); 43: 2183-2189). Therefore, without wishing to be bound by theory, since the functions of FABP3 are reported to include the transport of free fatty acids (see, e.g., Storch et al., Biochem. Biophys. Acta. 1486 (2000), 28-44), it can be assumed that the relevance of FABP3 to the effects induced by SGLTi is caused by the metabolic switch of SGLTi from carbohydrate to lipid utilization and ketogenesis.

[0021] Other markers commonly known to those skilled in the art related to cardiovascular disease, particularly heart failure, fail to detect the effect induced by SGLT2i. For example, growth differentiation factor-15 (GDF-15) is expressed by a variety of cell types, including cardiomyocytes, smooth muscle and endothelial muscle cells, and it is reported that the increase of plasma GDF-15 has a prognostic effect in heart failure, and it has also been found that it can be used to identify patients with heart failure who are responsive to the therapy comprising statins (see, e.g., Castiglione V, Aimo A, Vergaro G, Saccaro L, Passino C and Emdin M. Biomarkers for the diagnosis and management of heart failure. Heart Failure Reviews (2022) 27: 625-643). However, as demonstrated in the experimental section herein, in subjects treated with SGLT2i drugs, the amount of GDF-15 is substantially unchanged. In addition, the role of B-type NP (BNP) and N-terminal pro-B-type NP (NT-proBNP) in the diagnosis and risk stratification of heart failure has been reported (see, e.g., Castiglione V, Aimo A, Vergaro G, Saccaro L, Passino C and Emdin M. Biomarkers for the diagnosis and management of heart failure. Heart Failure Reviews (2022) 27: 625-643). However, as also demonstrated in the experimental section herein, NT-proBNP alone does not significantly change with SGLTi treatment, but as demonstrated in the experimental section, FABP3 used in combination with NT-proBNP can be reliably used to detect effects induced by SGLTi, wherein the ratio between the amount of FABP3 and the amount of NT-proBNP correlates with the effects induced by SGLTi. Thus, as demonstrated in the experimental section herein, markers known to be associated with cardiovascular disease (such as GDF-15 or NT-proBNP alone) do not indicate effects induced by SGLTi in subjects with cardiovascular disease. Surprisingly, the effect was detected by the amount of FABP3 or the association of FABP3 with NT-proBNP. Fatty acid binding proteins (FABPs) belong to a family of carrier proteins for fatty acids and other lipophilic substances. They are membrane-bound, facilitating the uptake of long-chain fatty acids into the cell, and are cytoplasmic, where they are crucial for the intracellular transport of fatty acids to sites of metabolic conversion. Consequently, FABPs are ubiquitous, and all nine FABP family members are distributed across various tissues.The FABP family includes liver (L-FABP / FABP1) isoforms, intestinal (I-FABP / FABP2) isoforms, cardiac (H-FABP / FABP3) isoforms, adipocyte (A-FABP / FABP4 / aP2) isoforms, epidermal (E-FABP / FABP5 / mal1) isoforms, ileal (Il-FABP / FABP6) isoforms, brain (B-FABP / FABP7) isoforms, myelin (M-FABP / FABP8) isoforms, and testicular (T-FABP / FABP9) isoforms, and due to their structural differences, each FABP member has different ligand selectivity and binding affinity for fatty acids (see, e.g., Furuhashi M, Hotamisligil GS: Fatty acid binding proteins: role in metabolic diseases and potential as drug targets. Nat Rev Drug Discov, 2008; 7: 489-503). For example, FABP4, also known as adipocyte FABP (A-FABP), is mainly expressed in adipocytes and macrophages and plays an important role in the development of insulin resistance and atherosclerosis, and its expression is reported to be associated with the pathogenesis of several diseases, mainly diabetes (see, for example, Furuhashi F. Fatty Acid-Binding Protein 4 in Cardiovascular and Metabolic Diseases. The official journal of the Japan Atherosclerosis Society and the Asian Pacific Society of Atherosclerosis and Vascular Diseases. 2019; 26: 216-232). In this regard, it has been reported that FABP4, but not FABP3, is positively correlated with the risk of diabetes, suggesting that the roles and functions of different FABPs in the FABP family are different (see, for example, Djousse L and Gaziano JM. Plasma levels of FABP4, but not of FABP3, are associated with increased risk of diabetes. Lipids (2012); 47 (8): 757-762).FABP3 is mainly expressed in cardiomyocytes and is also expressed in skeletal muscle, kidney, breast, testis, lung and stomach (see, e.g., Watanabe K, Wakabayashi H, Veerkamp JH et al., Immunohistochemical distribution of heart-type fatty acid-binding protein immunoreactivity in normal human tissues and in acute myocardial infarct. J Pathol. 1993; 170(1): 59–65; Zschiesche-Appie Kleine WH, Spitzer Jacques Veerkamp-Jan FC, Glatz EH et al., Histochemical localization of heart-type fatty-acidbinding protein in human and murine tissues. Histochem Cell Biol. 1995; 103(2): 147–156.). It has been reported that normal plasma levels of FABP3 in healthy human individuals are very low compared to levels of FABP3 in human plasma after acute cardiac injury and infarction (see, e.g., Kleine AH, Glatz JFC, Frans A, Van Nieuwenhoven, and Van der Vusse GJ. Release of heart fatty acid-binding protein into plasma after acute myocardial infarction in man. Molecular and Cellular Biochemistry 1992; 116: 155-162). Cardiac injury can be assessed based on the subject's general health and includes methods known in the art such as electrocardiogram, echocardiogram, angiography, or blood tests. In addition, cardiac injury can be assessed by determining the amount of cardiac injury markers such as NT-proBNP, GDF-15, or troponin. In some embodiments, it is envisioned that subjects with cardiovascular disease as disclosed herein do not suffer from cardiac injury, particularly acute cardiac injury, which would result in higher amounts of FABP3 compared to healthy individuals. Those skilled in the art know that acute cardiac injury results in higher FABP3 levels in plasma compared to healthy subjects and are able to determine acute cardiac injury in a subject.Thus, in subjects with cardiovascular disease and acute cardiac injury, one skilled in the art recognizes that the increased levels of FABP3 are caused by cardiac injury and can distinguish between the amount of FABP3 caused by damaged cells (i.e., cardiac injury) and the effects induced by SGLTi. In particular, one skilled in the art knows how to distinguish between the amount of FABP3 caused by damaged cells (i.e., cardiac injury) and the effects induced by SGLTi because one skilled in the art recognizes that the amount of FABP3 caused by the effects induced by SGLTi is associated with, for example, increased metabolic function, higher lipid turnover, better microcirculation as compared to a second marker of cardiac injury that is not involved in metabolic pathways, but only with structural proteins (such as troponin). For example, in a subject treated with SGLTi but who does not develop signs of acute cardiac injury (such as elevated troponin), an increase in the amount of FABP3 indicates that the increase in FABP3 is not caused by cardiac injury, but rather is caused by additional effects induced by SGLTi (such as altered metabolic function, lipid turnover, and / or better microcirculation). In addition, the time course of clinical signs and markers in a subject can be determined by methods known in the art and as disclosed herein, such as by an increase or decrease in the amount of a cardiac injury marker (such as NT-proBNP, GDF-15, or troponin), wherein an increase in the amount of a cardiac injury marker (such as NT-proBNP, GDF-15, or troponin) indicates worsening. To this end, a determination of worsening cardiac damage, for example, by an increase in a cardiac injury marker, can be distinguished by comparison with a change in the amount of FABP3. If the amount of FABP3 determined in the methods of the present invention exceeds the increase in the amount corresponding to worsening of the heart disease, the excess amount of FABP3 indicates an effect induced by SGLTi.

[0022] Thus, the amount of FABP3 and the worsening or improvement of cardiac damage can be monitored over time to track the relationship between the amount of FABP3 corresponding to the worsening or improvement of cardiac damage. Thus, in some embodiments of the methods of the present invention, the amount of FABP3 and, optionally, the amount of a cardiac damage marker is monitored over time to detect the effect induced by SGLTi. Preferably, the amount of FABP3 and, optionally, the amount of a cardiac damage marker is monitored for at least 1 hour, at least 1 day, at least 1 week, at least 2 weeks, or at least 4 weeks, more preferably 1 hour, 1 day, 1 week, 2 weeks, or 4 weeks. For example, in a subject suffering from a cardiovascular disease according to the present invention, the amount of FABP3 is monitored, wherein an amount of FABP3 greater than the amount of FABP3 corresponding to the worsening of cardiac damage (as determined by methods known in the art or by cardiac damage markers) indicates an effect induced by SGLTi. In this regard, it is understood by those skilled in the art that if the amount of FABP3 monitored is equal to or not greater than the amount of FABP3 corresponding to the worsening of cardiac damage, then this indicates that the amount of FABP3 is caused by cardiac damage. Thus, as disclosed above, determining and preferably monitoring the amount of FABP3 and cardiac damage, for example by determining markers for cardiac damage, simultaneously allows distinguishing the amount of FABP3 caused by SGLTi-induced effects from the amount of FABP3 caused by cardiac damage.

[0023] As used herein, the term "detecting" generally refers to detecting an effect induced by SGLTi using information or data related to the amount of FABP3 referred to herein in a sample from a subject. As used herein, the term "effect induced by SGLTi" refers to an effect caused by the action of SGLTi in a subject suffering from cardiovascular disease. In particular, as disclosed above, the effect is related to the amount of FABP3 in the subject, for example, the effect can be induced by SGLTi through a fasting-like metabolic paradigm involving a metabolic switch from carbohydrate to lipid utilization and ketogenesis. As also disclosed herein, the term "detecting" can also be viewed as "visualizing" or "assessing" an effect induced by SGLTi in a subject suffering from cardiovascular disease. As previously described - for example, FABP3 can be used to assess / visualize whether SGLTi is already active in a subject suffering from cardiovascular disease; or is still active in a subject suffering from cardiovascular disease; or is fully active in the subject suffering from cardiovascular disease, etc.

[0024] According to the above method of the present invention, it is envisaged that the subject should be a subject suffering from cardiovascular disease. The terms "subject" or "patient" as used interchangeably herein may relate to animals, preferably mammals, and more preferably humans. In particular, the terms "patient" or "subject" as used herein may be any single human subject who is experiencing or has experienced one or more signs, symptoms or other indicators of cardiovascular disease, particularly heart failure, that meet treatment conditions. The subject should exhibit symptoms associated with this, i.e., at least be suspected of having cardiovascular disease. As used herein and according to the present invention, " cardiovascular disease " relates to any clinical manifestation of the disease state relevant to the heart, heart valve and vascular system (for example, vein and artery) of health, and covers disease and illness, including but not limited to heart failure, particularly chronic heart failure, arteriosclerosis, atherosclerosis, coronary heart disease, cardiomyopathy, myocardial infarction, acute coronary syndrome, angina pectoris, aortic aneurysm, aortic dissection, iliac artery aneurysm or femoral artery aneurysm, pulmonary embolism, essential hypertension, atrial fibrillation, stroke, transient ischemic attack, systolic dysfunction, diastolic dysfunction, myocarditis, atrial tachycardia, ventricular fibrillation, endocarditis, peripheral vascular disease, coronary artery disease (CAD), peripheral artery disease (PAD) and cerebrovascular disease.In some instances, the subject suffering from cardiovascular disease also may suffer from diabetes, particularly type 2 diabetes.In certain embodiments, the subject suffering from cardiovascular disease in the context of the present invention is a subject suffering from heart failure. As used herein, the term "heart failure" is a disease that can be caused by any structural or functional heart disease, which impairs the ability of the heart to fill or pump a sufficient amount of blood throughout the body. The signs and symptoms of heart failure are well known in the art. In particular, as known to those skilled in the art, heart failure relates to diastolic dysfunction, or in particular, to systolic dysfunction of the heart with obvious signs of heart failure. Preferably, the heart failure mentioned herein is chronic heart failure (which is preferably caused by systolic dysfunction). As used herein, the term "chronic heart failure" refers to chronic, i.e. permanent heart failure. Heart failure is characterized by impaired diastolic or systolic blood flow rate, and therefore impaired cardiac function. However, the chronic heart failure mentioned herein is not a sudden ischemia with severe necrosis of myocardial cells, but is preferably accompanied by persistent necrotic events in myocardial cells, thereby causing the continued development of cardiac function to be impaired. In certain embodiments, the subject suffers from systolic or diastolic heart failure. Heart failure according to the present invention includes obvious and / or late heart failure. In obvious heart failure, the patient shows heart failure symptoms known to those skilled in the art.Heart failure can be classified into different degrees of severity.According to the NYHA (New York Heart Association) classification, heart failure patients are classified as belonging to NYHA Class I, Class II, Class III and Class IV.Patients with NYHA class I have no overt symptoms of cardiovascular disease but have objective evidence of impaired function. Patients with NYHA class II have mild limitations in physical activity. Patients with NYHA class III exhibit significant limitations in physical activity. Patients with NYHA class IV are unable to perform any physical activity without discomfort. They exhibit symptoms of cardiac dysfunction at rest. This functional classification is supplemented by the latest classification from the American College of Cardiology and the American Heart Association (see J.Am.Coll.Cardiol. 2001;38;2101-2113, updated in 2005, see J.Am.Coll.Cardiol. 2005;46;el-e82). Four stages, A, B, C, and D, are defined. Stage A and stage B patients are best defined as those with risk factors for developing heart failure. For example, patients with coronary artery disease, hypertension, or diabetes who do not yet demonstrate impaired left ventricular (LV) function, hypertrophy, or geometric malformation would be considered stage A, while patients who are asymptomatic but demonstrate LV hypertrophy and / or impaired LV function would be designated stage B. Stage C indicates patients with current or past symptoms of heart failure associated with underlying structural heart disease, while stage D indicates patients with truly refractory heart failure. In some embodiments, the subject with cardiovascular disease has heart failure classified as stage B, C, or D according to the ACC / AHA classification, and / or wherein the subject has heart failure of class II to IV according to the NYHA classification. In some embodiments, the subject with cardiovascular disease has heart failure with impaired left ventricular ejection fraction (LVEF). Heart failure can be divided into the following categories based on left ventricular ejection fraction (LVEF): heart failure with preserved ejection fraction (HFpEF; LVEF ≥ 50%), heart failure with intermediate ejection fraction (HFmrEF; LVEF 40 to 49%), and heart failure with reduced ejection fraction (HFrEF; LVEF < 40%). HFpEF is characterized by diastolic dysfunction and is usually caused by myocardial damage caused by comorbidities (e.g., obesity, chronic kidney disease (CKD), chronic obstructive pulmonary disease) or accumulation diseases (e.g., cardiac amyloidosis). HFrEF, on the other hand, is characterized by systolic dysfunction, secondary to direct cardiac insult (such as acute coronary syndrome), cardiomyopathy, or valvular disease. Thus, in some instances, a subject may have heart failure with reduced ejection fraction (HFrEF), mildly reduced ejection fraction (HRMEF), or preserved ejection fraction (HFpEF).

[0025] As used herein, the term "sample" refers to a body fluid sample, a separated cell sample, or a tissue or organ sample. The sample of body fluid can be obtained by well-known technology. Tissue or organ samples can be obtained from any tissue or organ by, for example, biopsy. Separated cells can be obtained from body fluids, tissues, or organs by separation techniques such as centrifugation or cell sorting. Preferably, cells, tissues, or organ samples are obtained from those cells, tissues, or organs that express or produce the polypeptides mentioned herein. The sample can be a frozen sample, a fresh sample, a fixed (e.g., formalin fixed) sample, a centrifugal and / or embedded (e.g., paraffin embedded) sample, etc. Before assessing the amount of one or more markers in the sample, the cell sample can certainly be subjected to various well-known post-collection preparation and storage techniques (e.g., nucleic acid and / or protein extraction, fixation, storage, freezing, ultrafiltration, concentration, evaporation, centrifugation, etc.). Similarly, biopsy specimens can also be processed by post-collection preparation and storage techniques (e.g., fixation). In certain embodiments, the sample is a blood, serum, plasma, or urine sample, and more preferably, the sample is a blood, serum, or plasma sample. In particular, it is contemplated to measure the levels of biomarkers in plasma samples.

[0026] In the context of the present invention, the level or amount of FABP3 is determined in a sample from a subject suffering from cardiovascular disease. Additionally, in some embodiments, the amount of a cardiac injury marker is determined. Thus, the amount of a single marker can be measured, or a combination of markers comprising the marker FABP3. In all embodiments of the present invention, the amount / level of the corresponding marker (e.g., biomarker) or polypeptide (FABP3, cardiac injury marker) used therein is determined by methods known to those skilled in the art. Preferably, the amount of a human marker or polypeptide is determined. Determining the amount of a corresponding marker or polypeptide as referred to herein involves measuring the amount or concentration, preferably in a semi-quantitative or quantitative manner. The terms "measuring," "detecting," and "determining" are used interchangeably herein and relate to the quantification of a marker or polypeptide, for example, to determining the level of a marker or polypeptide in a sample using an appropriate detection method as described herein. Measurement can be performed directly or indirectly. Direct measurement involves measuring the amount or concentration of a marker or polypeptide based on a signal obtained from the marker or polypeptide itself, and the intensity of the signal is directly related to the number of molecules of the marker or polypeptide present in the sample. Such a signal, sometimes referred to herein as an intensity signal, can be obtained, for example, by measuring the intensity value of a specific physical or chemical property of the marker or polypeptide. Indirect measurements include measuring a signal obtained from a secondary component (i.e., a component other than the marker or polypeptide itself) or a biological readout system (e.g., a measurable cellular response, ligand, label, or enzymatic reaction product).

[0027] According to the present invention, determining the amount of a marker or polypeptide can be achieved by any suitable tool and method for determining the amount of a marker or polypeptide in a sample. Said tool and method include immunoassay devices and methods, which can utilize various sandwich, competitive or other assay formats of labeled molecules. Those skilled in the art generally know which of the following methods are suitable for qualitative and / or quantitative detection of biomarkers. Commercially available Western and immunoassays such as ELISA, RI-As, and fluorescence-based immunoassays can be used to conveniently measure, for example, proteins in a sample. Other suitable methods include measuring the unique physical or chemical properties of a marker or polypeptide, such as its accurate molecular mass or NMR spectrum. Said method includes, preferably, a biosensor, an optical device coupled to an immunoassay, a biochip, an analytical device (such as a mass spectrometer, an NMR analyzer, or a chromatographic device). Further, the method includes a method based on microplate ELISA, fully automatic or robotic immunoassays (e.g., available on an Elecsys analyzer), CBA (e.g., available on a Roche-Hitachi TM Enzyme cobalt binding assays available on analyzers) and latex agglutination assays (e.g., on Roche-Hitachi TM Analyzer available). Preferably, determining the amount of a marker or polypeptide comprises the following steps: (a) contacting a cell capable of eliciting a cellular response with the marker or polypeptide for a sufficient period of time, the intensity of the cellular response being indicative of the amount of the marker or polypeptide, and (b) measuring the cellular response. To measure the cellular response, the sample or treated sample is preferably added to a cell culture and an internal or external cellular response is measured. The cellular response may include measurable expression of a reporter gene or secretion of a substance (e.g., a peptide, polypeptide, or small molecule). The expression or substance should produce an intensity signal that is correlated with the amount of the marker or polypeptide. Also preferably, determining the amount of a marker or polypeptide comprises the step of measuring a specific intensity signal that can be obtained from the marker or polypeptide in the sample. As described above, such a signal may be the signal intensity observed at an m / z variable specific to the peptide or polypeptide observed in a mass spectrometer or an NMR spectrum specific to the peptide or polypeptide. Determining the amount of a marker or polypeptide may preferably comprise the following steps: (a) contacting the marker or polypeptide with a specific ligand, (b) (optionally) removing unbound ligand, and (c) measuring the amount of bound ligand. The bound ligand will generate an intensity signal. Binding according to the present invention includes covalent and non-covalent binding. The ligand according to the present invention can be any compound that is bound to a peptide or polypeptide as described herein, for example, a peptide, polypeptide, nucleic acid or small molecule. Methods for preparing such ligands are well known in the art.

[0028] The amount of the marker or polypeptide can also be preferably determined as follows: (a) contacting a ligand for the marker or polypeptide as specified above with a sample comprising the marker or polypeptide and (b) measuring the amount of the marker or polypeptide bound to the support. The ligand preferably selected from the group consisting of the following is preferably present on a solid support in an immobilized form: nucleic acids, peptides, polypeptides, antibodies and aptamers. The materials used to make solid supports are well known in the art and include, in particular, commercially available column materials, polystyrene beads, latex beads, magnetic beads, colloidal metal particles, glass and / or silicon wafers and surfaces, nitrocellulose tapes, membranes, sheets, duracytes, holes and walls of reaction trays, plastic tubing, etc. The ligand or agent can be bound to many different carriers. Examples of well-known carriers include glass, polystyrene, polyvinyl chloride, polypropylene, polyethylene, polycarbonate, dextran, nylon, amylose, natural and modified cellulose, polyacrylamide, agarose and magnetite. For the purposes of the present invention, the nature of the carrier can be soluble or insoluble. Suitable methods for fixing / immobilizing the ligand are well known and include, but are not limited to, ionic, hydrophobic, covalent interactions, and the like.

[0029] As used herein, the term "amount" encompasses the absolute amount of a marker (such as FABP3 or NT-proBNP) mentioned herein, the relative amount or concentration of the biomarker, and any value or parameter related thereto or derivable therefrom. Such values ​​or parameters include intensity signal values ​​from all specific physical or chemical properties obtained from the marker by direct measurement, such as intensity values ​​from a mass spectrometer or NMR spectrum. In addition, included are all values ​​or parameters obtained by indirect measurement as described herein, for example, the amount of reaction determined from a biological readout system in response to an intensity signal obtained from a peptide or a specifically bound ligand. It should be understood that values ​​related to the above-mentioned amounts or parameters can also be obtained by all standard mathematical operations.

[0030] As used herein, the term "comparison" refers to comparing the amount of a marker (such as FABP3 or NT-proBNP) in a sample from a subject or patient with a reference amount of a biomarker described herein. It should be understood that comparison, as used herein, generally refers to the comparison of corresponding parameters or values, for example, comparing an absolute amount with an absolute reference amount, comparing a concentration with a reference concentration, or comparing an intensity signal obtained from a marker in a sample with an intensity signal of the same type obtained from a reference sample. Comparisons can be performed manually or with computer assistance. Thus, comparisons can be performed by a computing device. For example, the value of the measured or detected amount of a biomarker in a sample from an individual or patient can be compared to a reference level, and the comparison can be performed automatically by a computer program executing a comparison algorithm. The computer program performing the evaluation will provide the desired assessment in an appropriate output format. For computer-assisted comparisons, the value of the determined amount can be compared to a value corresponding to an appropriate reference stored in a database by the computer program. The computer program can further evaluate the results of the comparison, i.e., automatically provide the desired assessment in an appropriate output format. For computer-assisted comparisons, the value of the determined amount can be compared to a value corresponding to an appropriate reference stored in a database by the computer program. The computer program can further evaluate the results of the comparison, i.e., automatically provide the desired evaluation in a suitable output format. Based on the comparison of the amount determined in step a) with a reference amount, the effect induced by SGLTi can be detected. Therefore, the reference amount is selected such that the difference or similarity of the comparison amount allows the detection of the effect induced by SGLTi according to the present invention.

[0031] As used herein and in the context of the present invention, a "reference amount" preferably refers to a determined or predetermined value obtained from a sample of a subject or from a sample of a group of subjects suffering from cardiovascular disease. Preferably, the reference amount is determined or predetermined in a reference sample from the disease entity to which the patient belongs. Thus, the reference amount will typically be from a subject known to suffer from cardiovascular disease. For example, if the subject suffers from heart failure, the reference amount is determined or predetermined in a sample from a subject who is also expected to suffer from heart failure. It is expected that the extent of cardiovascular disease (such as heart failure) in the subject is substantially the same as that in the reference subject. The extent or severity of cardiovascular disease (such as heart failure) can be determined without further elaboration by one skilled in the art and is also disclosed above. Such a reference amount can be, for example, a threshold amount, wherein an amount greater than the threshold value is indicative of an effect induced by SGLTi. In some instances, the reference amount refers to the amount of FABP3 before the subject is treated with SGLTi. In particular, in some instances, the reference amount of FABP3 before the subject is treated with SGL of FABP3 is approximately 37.8 ng / mL. Preferably, the term "about" as used herein includes a range of + and -20% relative to a specific amount, more preferably a range of + and -10%, even more preferably a range of + and -5%, and most preferably a range of + and -2%, for example, an amount representing "about 100" means an amount within a range of 80 to 120. In addition, the term "about" refers to an exact amount. Preferably, the amount or level is measured as described in the examples. In some embodiments, the reference amount is the upper limit of normal (ULN) of the amount in a sample from a subject with cardiovascular disease. The ULN for a sample from a subject, i.e., a given subject population, can be determined by various well-known techniques. In this case, those skilled in the art know how to set the reference amount to meet conventional requirements. The reference amount can, for example, be set to any percentage between 25% and 75%, or to a median, tertile, or quartile as determined from the overall distribution of values ​​in the reference sample to be determined. Typically, the reference sample is derived from substantially the same type of cell, tissue, organ or bodily fluid source as the sample from the subject or patient subjected to the method of the invention, e.g., if blood is used as a sample to determine the level of a biomarker in an individual according to the invention, the reference amount is also determined in the blood or a fraction thereof.

[0032] In some examples, the reference amount can include a reference amount of FABP3 and a reference amount of a cardiac injury marker. In particular, in some embodiments, the method comprises generating a ratio between the amount of FABP3 and the amount of a cardiac injury marker and comparing the ratio to a reference ratio. For example, the cardiac injury marker is a cardiac function marker, a myocardial necrosis marker, or a cardiac inflammation marker. The threshold reference amount or reference ratio can be calculated without further elaboration by statistical testing in samples from subjects with cardiovascular disease and / or from a group of subjects with cardiovascular disease. For example, the reference amount can be determined based on clinical studies and from a receiver operating characteristic (ROC) curve as described herein. For example, the reference amount can be determined in reference samples from a subject or a group of subjects with cardiovascular disease (such as heart failure) who were treated or not treated with SGLTi. For example, the reference amount can be determined in a subject treated with SGLTi for monitoring an effect induced by SGLTi. As another example, the reference amount can be determined in a subject not treated with SGLTi, for example, to quantify an effect induced by SGLTi in the subject. In some embodiments, the reference amount is the amount in a sample from a subject with cardiovascular disease or the ULN of the amount in a sample from a subject with cardiovascular disease prior to treatment with SGLTi. In particular, it is contemplated that the subject and a reference subject, or a group thereof, have substantially the same reference amount of FABP3. One skilled in the art can readily assess whether two amounts are substantially the same. For example, amounts that differ by less than 5% or less than 3% are considered substantially the same.

[0033] In some embodiments, the sodium-glucose co-transporter inhibitor (SGLTi) is a sodium-glucose co-transporter 2 inhibitor (SGLT2i) or a sodium-glucose co-transporter 1 inhibitor (SGLT1i), preferably an SGLT2i, more preferably an SGLT2i, more preferably a gliflozin, more preferably the SGLTi comprises canagliflozin, dapagliflozin and empagliflozin, ertugliflozin, ipragliflozin, remogliflozin, sergliflozin, sotagliflozin or tofogliflozin. Currently available SGLT inhibitors are developed based on the natural product phlorizin and include, for example, but not limited to, SGLT2 inhibitors such as empagliflozin, dapagliflozin, and canagliflozin; SGLT1 inhibitors such as KGA-2727 and mizagliflozin; and dual inhibitors that inhibit SGLT1 and SGLT2 such as sogliflozin and LX2761. In particular, sodium-glucose cotransporter 2 (SGLT2) is a high-capacity, low-affinity cotransporter located in the proximal tubular epithelium of the kidney and primarily reabsorbs filtered glucose (Bakris GL, Fonseca VA, Sharma K, Wright EM. Renalsodium-glucose transport: role in diabetes mellitus and potential clinical implications. Kidney international. 2009; 75(12): 1272-7). In addition to its renal-specific effects on glycemic control, SGLT2 is normally responsible for ≈5% of sodium reabsorption in the proximal tubule. In the setting of chronic hyperglycemia, the capacity of SGLT2 in the kidney is increased, resulting in a more pronounced effect on sodium homeostasis (Bakris GL, Fonseca VA, Sharma K, Wright EM. Renal sodium-glucose transport: role in diabetes mellitus and potential clinical implications. Kidney international. 2009; 75(12): 1272-7).Subsequently, inhibition of SGLT2 increases diuresis and reduces plasma volume and blood pressure (Baker WL, Smyth LR, Riche DM, Bourret EM, Chamberlin KW, White WB. Effects of sodium-glucose co-transporter2inhibitors on blood pressure: a systematic review and metaanalysis. Journal of the American Society of Hypertension: JASH. 2014; 8 (4): 262-75.e9). SGLT2i has been shown to exert cardioprotective and renoprotective effects by promoting autophagic flux and alleviating apoptosis, inhibiting oxidative and other cellular stresses, inhibiting proinflammatory and fibrotic pathways, and enhancing cellular energy storage and metabolism. Thus, it has been reported that SGLT2i improves the cardiovascular outcomes of patients, including patients with heart failure, such as heart failure with reduced or preserved ejection fraction. The methods of the present invention contemplate administering SGLT2i to subjects with cardiovascular disease. The term "administer" as used herein is used in the broadest sense and particularly includes oral, enteral, topical, and "parenteral administration." As used herein, "parenteral administration" and "administered parenterally" refer to administration other than enteral and topical administration (usually by injection), and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, intrasternal injection, and infusion.

[0034] As used herein, the term "FABP3" refers to cardiac fatty acid binding protein, particularly human cardiac fatty acid binding protein. Preferably, the term also includes variants of cardiac fatty acid binding protein. FABP3 is often also referred to as cardiac fatty acid binding protein. FABP3 is also known as H-FABP. The cDNA sequence and protein sequence of human FABP3 are well known in the art and were originally described by Peters et al. (Biochem. J. 276 (Pt 1), 203-207 (1991)). In addition, the sequence of human H-FABP can preferably be found in Genebank entries U57623.1 (cDNA sequence) and AAB02555.1 (protein sequence). The main physiological function of FABP3 is believed to be the transport of free fatty acids, see, for example, Storch et al., Biochem. Biophys. Acta. 1486 (2000), 28-44. As used herein, FABP3 also encompasses variants of the aforementioned specific FABP3 polypeptides. Such variants have at least the same basic biological properties as the specific FABP3 polypeptide. In particular, they share the same basic biological properties if they can be detected by the same specific assays mentioned in this specification, such as by an ELISA assay using a polyclonal or monoclonal antibody that specifically recognizes the FABP3 polypeptide. Preferred assays are described in the accompanying Examples. Furthermore, it is understood that variants according to the present invention have amino acid sequences that differ by at least one amino acid substitution, deletion, and / or addition, wherein the amino acid sequence of the variant is still preferably at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 92%, at least about 95%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence of the specific FABP3 polypeptide, preferably to the amino acid sequence of human FABP3, more preferably over the entire length of the specific FABP3 (e.g., human FABP3). The degree of identity between two amino acid sequences can be determined as described above. The variants mentioned above may be allelic variants or any other species-specific homologs, paralogs, or orthologs. In addition, the variants mentioned herein include fragments of specific FABP3 polypeptides or variants of the aforementioned types, as long as these fragments have the basic biological properties mentioned above. Such fragments may be, for example, degradation products of FABP3 polypeptides. Variants that differ due to post-translational modifications (such as phosphorylation or myristylation) are also included.

[0035] In some embodiments, the present invention further comprises additionally determining the amount of a cardiac injury marker. As used herein, a cardiac injury marker indicates myocardial damage, injury, or necrosis, for example, where muscle mass is reduced, meaning the heart's contractile ability is reduced. In some instances, the cardiac injury marker is preferably a marker of cardiac function, cardiac inflammation, or myocardial necrosis. Preferred cardiac injury markers are cardiac function markers such as NT-proBNP or myocardial necrosis markers such as cardiac troponin. In some instances, an increase in the amount of FABP3 compared to a reference amount and no increase in the amount of a cardiac injury marker indicate an effect induced by SGLTi. One skilled in the art can readily assess whether a cardiac injury marker is increased. For example, an amount that differs by less than 5% or less than 3% is considered not increased. As used herein, a cardiac function marker indicates myocardial dysfunction, i.e., muscle tissue in the myocardium is weaker than normal tissue and cannot contract as well as healthy tissue, meaning the heart must work harder than normal tissue to ensure adequate blood supply to the body. As used herein, a myocardial necrosis marker indicates cell death that has occurred in the subject's myocardium, which may occur after a prolonged ischemic state or as a result of apoptosis. As used herein, an inflammatory marker indicates an inflammatory process occurring in an individual, particularly in the myocardium. In one embodiment, the cardiac marker is a cardiac function marker, preferably wherein the cardiac function marker is a natriuretic peptide, more preferably BNP or NT-proBNP, more preferably NT-proBNP. The term "natriuretic peptide" includes atrial natriuretic peptide (ANP)-type and brain natriuretic peptide (BNP)-type peptides and their variants. BNP-type peptides include pre-proBNP, proBNP, NT-proBNP and BNP (see, e.g., Bonow, 1996, Circulation 93: 1946-1950; WO 02 / 089657, WO 02 / 083913). The pre-pro peptide of the aforementioned brain natriuretic peptide (134 amino acids in length) includes a short signal peptide that is enzymatically cleaved to release the pro peptide (108 amino acids). The leader peptide is further cleaved into an N-terminal leader peptide (NT-pro peptide, 76 amino acids) and an active hormone (32 amino acids). Preferably, the brain natriuretic peptide according to the present invention is BNP, NT-proBNP, and variants thereof. The most preferred BNP-type peptide mentioned herein is human NT-proBNP. Conventional diagnostic techniques for heart failure are based on the well-known vascular volume stress marker NT-proBNP. As briefly discussed above, human NT-proBNP according to the present invention is a polypeptide preferably comprising 76 amino acids corresponding to the N-terminal portion of the human NT-proBNP molecule.The structures of human BNP and NT-proBNP are described in detail in the prior art, for example, in WO 02 / 089657 and WO 02 / 083913. Preferably, human NT-proBNP as used herein is human NT-proBNP as disclosed in EP 0 648 228 B1. Reference to NT-proBNP according to the present invention further encompasses alleles and other variants of the specific sequence of human NT-proBNP discussed above. Specifically, variant polypeptides that are at least 60%, more preferably at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or at least 99% identical to human NT-proBNP at the amino acid level are contemplated. Proteolytic degradation products are generally similar and are also contemplated, and are still recognized by identification means or by ligands directed against the corresponding full-length peptide. Variant polypeptides having amino acid deletions, substitutions, and / or additions compared to the amino acid sequence of human NT-proBNP are also contemplated, provided that the polypeptide possesses the properties of NT-proBNP. The NT-proBNP properties referred to herein are immunological and / or biological properties. Preferably, NT-proBNP variants have immunological properties (i.e., epitope composition) comparable to those of NT-proBNP. Thus, the variants should be identifiable by the aforementioned means or ligands for determining the amount of natriuretic peptides. Biological and / or immunological NT-proBNP properties can be detected by the assays described by Karl et al. (Karl 1999, Scand J Clin Invest 59:177-181) and Yeo et al. (Yeo 2003, Clinica Chimica Acta 338:107-115). Variants also include post-translationally modified peptides, such as glycosylated peptides. Furthermore, variants according to the present invention are peptides or polypeptides that have been modified after sample collection, for example, by covalently or non-covalently attaching a label, particularly a radioactive or fluorescent label, to the peptide. For example, a marker of myocardial necrosis can be cardiac troponin. As used herein, the term "cardiac troponin" refers to all troponin isoforms expressed in cardiac cells. These subtypes are well characterized in the prior art as described, for example, in Anderson 1995, Circulation Research, Vol. 76, No. 4: 681-686 and Ferrieres 1998, Clinical Chemistry, 44: 487-493. Patients suffering from myocardial infarction can be diagnosed using cardiac troponins, preferably troponin T or I. Myocardial infarction is believed to be caused by a necrotic state (i.e., cell death) of the myocardium. Cardiac troponins are released after cell death and can therefore be used to diagnose myocardial infarction. If the amount of troponin T in the blood is elevated, i.e., above 0.1 ng / ml, an acute cardiovascular event is assumed and the patient is treated accordingly.Preferably, cardiac troponin refers to troponin T or troponin I. It should be understood that the subtypes of troponin can be determined together in the method of the present invention, i.e., simultaneously or sequentially, or individually, i.e., the other subtypes are not determined at all. The amino acid sequences for human troponin T and human troponin I are disclosed in Anderson, Loc Cit and Ferrieres 1998, Clinical Chemistry, 44: 487-493. The term "cardiac troponin" also encompasses variants of the aforementioned specific troponins. Such variants have at least the same basic biological properties and immunological properties as the specific cardiac troponin. In particular, if they can be detected by the same specific assays mentioned in this specification, such as by ELISA assays using polyclonal antibodies or monoclonal antibodies that specifically recognize the cardiac troponin, they share the same basic biological properties and immunological properties. Furthermore, it is understood that variants as referred to herein should have an amino acid sequence that differs due to at least one amino acid substitution, deletion and / or addition, wherein the amino acid sequence of the variant is still preferably at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 92%, at least about 95%, at least about 97%, at least about 98% or at least about 99% identical to the amino acid sequence of the specific troponin. Variants may be allelic variants or any other species-specific homologs, paralogs or orthologs. Furthermore, variants as referred to herein include fragments of the specific cardiac troponin or variants of the aforementioned types, as long as these fragments have the basic immunological and biological properties as mentioned above. Preferably, the cardiac troponin variant has immunological properties (i.e., epitope composition) comparable to those of human troponin T or troponin I. Thus, the variant should be recognizable by the above-mentioned means or ligands for determining the amount of cardiac troponin. Therefore, the variant should be identifiable by the above-mentioned means or ligand for determining the amount of cardiac troponin. Such fragments can be, for example, degradation products of troponin. Variants that are different due to post-translational modifications (such as phosphorylation or myristylation) are also included. Preferably, the biological properties of troponin I and its variants are the ability to inhibit actomyosin ATPase or angiogenesis in vivo and in vitro, which can be detected by the assay method described by Moses et al. 1999 PNAS USA 96 (6): 2645-2650). Preferably, the biological properties of troponin T and its variants are the ability to form a complex with troponin C and I, bind calcium ions or bind myosin, preferably, if present as a complex of troponin C, I and T or a complex formed by troponin C, troponin I and troponin T variants.Preferably, the amount of cardiac troponin, in particular troponin T, is determined using a very sensitive troponin T assay system, so that very low amounts of cardiac troponin can be reliably determined, preferably said assay system being capable of determining an amount of troponin of 0.002 ng / ml in a sample, preferably a blood, serum or plasma sample. A particularly preferred troponin T assay is 2010 analyzer (Roche Diagnostics) with a detection limit of about 0.001 ng / ml to about 0.0015 ng / ml, typically about 0.0015 ng / ml. The term "growth differentiation factor-15" or "GDF-15" refers to a polypeptide that is a member of the transforming growth factor (TGF) cytokine superfamily. GDF-15 was originally cloned as macrophage-inhibitory cytokine 1 and was later identified as placental transforming growth factor-15, placental bone morphogenetic protein, nonsteroidal anti-inflammatory drug-activated gene 1, and prostate-derived factor (Bootcov loc cit; Hromas, 1997 Biochim Biophys Acta 1354:40-44; Lawton 1997, Gene 203:17-26; Yokoyama-Kobayashi 1997, J Biochem (Tokyo), 122:622-626; Paralkar 1998, J Biol Chem 273:13760-13767). Similar to other TGF-related cytokines, GDF-15 is synthesized as an inactive precursor protein that undergoes disulfide-linked homodimerization. After proteolytic cleavage of the N-terminal propeptide, GDF-15 is secreted as an approximately 28 kDa dimeric protein (Bauskin 2000, Embo J 19:2212-2220). The amino acid sequence of GDF-15 is disclosed in WO99 / 06445, WO00 / 70051, WO2005 / 113585, Bottner 1999, Gene 237:105-111, Bootcov loc.cit, Tan loc.cit., Baek 2001, Mol Pharmacol 59:901-908, Hromas loc cit, Paralkar loc cit, Morrish 1996, Placenta 17:431-441 or Yokoyama Kobayashi loc cit.

[0036] The present invention also relates to a computer-implemented method for performing the method according to any of the preceding claims, the computer-implemented method comprising: a) means for determining the amount of FABP3, or the amount of FABP3 and a cardiac injury marker, in a sample from a subject treated with SGLTi, wherein the subject suffers from cardiovascular disease; and b) means for comparing the amount of FABP3 to a reference amount, comparing the amount of FABP3 and the amount of the cardiac injury marker to a reference amount, or generating a ratio between the amount of FABP3 and the amount of the cardiac injury marker and comparing the ratio to a reference ratio. It will be appreciated that at least some steps of the methods of the present invention can be performed by a computing device. The computing device can also access an output device. Exemplary output devices include, for example, a fax machine, a display, a printer, and a file. According to some embodiments of the present disclosure, a computing device can perform one or more steps of the methods disclosed herein and thereafter provide output related to the results, indications, ratios, or other factors of the methods via the output device. It should also be understood that multiple computing devices can be used together, such as over a network or through other methods of transmitting data, to perform one or more steps of the methods disclosed herein. Exemplary computing devices include desktop computers, laptop computers, cellular devices, tablet computers, servers, etc. Generally speaking, a computing device includes a processor capable of executing a plurality of instructions, such as programs of software.

[0037] The present invention also relates to a computer program product comprising program code for executing a computer-implemented method according to the present invention when executed on at least one computer. As used herein, software may include instructions that, when executed by a processor of a computing device, may perform one or more steps of the method disclosed herein. Some instructions may be suitable for generating signals that control the operation of other machines and may therefore be operable via these control signals to convert material away from the computer itself. These descriptions and representations are a means by which those skilled in the art of data processing, for example, can most effectively convey the content of their work to others skilled in the art.

[0038] The present invention also relates to a device for performing the method according to any of the preceding claims, comprising: a) means for determining the amount of FABP3 or the amount of FABP3 and a cardiac damage marker in a sample from a subject treated with SGLTi, wherein the subject suffers from cardiovascular disease; and b) means for comparing the amount of FABP3 to a reference amount, comparing the amount of FABP3 and the amount of the cardiac damage marker to a reference amount, or generating a ratio between the amount of FABP3 and the amount of the cardiac marker and comparing this ratio to a reference ratio. As used herein, the term "device" refers to a system comprising at least means operatively linked to each other to allow detection of an effect induced by SGLTi. Preferred means for determining the amount of FABP3 and, optionally, the amount of a cardiac damage marker (such as NT-proBNP), as well as means for performing the comparison, are disclosed above in conjunction with the method of the present invention. How the means are operatively linked will depend on the type of means included in the device. For example, in the case of using means for automatically determining the amount of a biomarker or polypeptide, the data obtained by the automated means can be processed, for example, by a computer program to obtain the desired results. Preferably, in this case, the tool consists of a single device. Therefore, the device may include an analytical unit for measuring the amount of peptides or polypeptides in the applied sample and a computer unit for processing the obtained data for evaluation. The computer unit preferably includes a database comprising a stored reference amount or its value described elsewhere in this specification, and a computer-implemented algorithm for comparing the determined amount of the polypeptide with the stored reference amount of the database. As used herein, computer-implemented refers to a computer-readable program code tangibly contained in the computer unit. Alternatively, when using a tool such as a test strip to determine the amount of a biomarker or polypeptide, the tool for comparison may include a control strip or table that assigns the determined amount to the reference amount. The test strip is preferably coupled to a ligand that specifically binds to the marker or polypeptide mentioned herein. The strip or device preferably includes a tool for detecting the binding of the marker or polypeptide to the ligand. Preferred detection tools are disclosed in conjunction with the embodiments related to the above-mentioned method of the present invention. In this case, the tool is operably connected because the user of the system brings together the determination results of the amount and its diagnostic or prognostic value due to the instructions and explanations given in the manual. In such embodiments, the tools may be provided as separate devices and preferably packaged together as a kit. One skilled in the art will readily appreciate how to connect these tools. Preferred devices are systems that require no specialized knowledge from a clinician, such as test strips or electronic devices that require only sample loading. Results may be output as raw data that require interpretation by a clinician.Preferably, the output of the device is processed raw data, i.e., evaluated raw data, the interpretation of which does not require a clinician. Other preferred devices include an analysis unit / device (e.g., a biosensor, an array, a solid support coupled to a ligand that specifically recognizes a natriuretic peptide, a plasma surface resonance device, an NMR spectrometer, a mass spectrometer, etc.) and / or an evaluation unit / device as mentioned above according to the method of the invention.

[0039] The present invention also relates to a kit for performing the method according to any of the preceding claims, comprising: a) means for determining the amount of FABP3 or the amount of FABP3 and a cardiac damage marker in a sample from a subject treated with SGLTi, wherein the subject suffers from cardiovascular disease; and b) means for comparing the amount of FABP3 to a reference amount, comparing the amount of FABP3 and the amount of the cardiac damage marker to a reference amount, or generating a ratio between the amount of FABP3 and the amount of the cardiac marker and comparing this ratio to a reference ratio. As used herein, the term "kit" refers to a collection of the aforementioned means, preferably provided separately or in a single container. The container preferably also includes instructions for performing the method of the present invention. These instructions may be in the form of a manual or may be provided by computer program code capable of performing the comparisons mentioned in the method of the present invention and, when implemented on a computer or data processing device, creating corresponding identifiers. In addition, the kit should include at least one reference standard as defined above, i.e., a solution having a reference amount of FABP3 and, optionally, other markers representing the reference amount, such as NT-proBNP as mentioned herein.

[0040] The present invention also relates to i) the use of FABP3, FABP3, and cardiac damage markers and / or ii) the use of detection agents for FABP3, FABP3, and cardiac damage markers for detecting effects induced by SGLTi in subjects with cardiovascular disease.

[0041] The present invention will be further illustrated by the following non-limiting experimental examples.

[0042] Experimental example

[0043] Example 1:

[0044] Study design and ethical aspects

[0045] This study was a prospective, single-center clinical trial conducted at the Department of Internal Medicine II / Division of Cardiology, Medical University of Vienna. The study protocol conformed to the Declaration of Helsinki and was approved by the Ethics Committee of the Medical University of Vienna (EK1729 / 2013).

[0046] All patients provided written informed consent before enrollment. Patient data collected in this study were anonymized. All personally identifiable information was removed or replaced in both the original and analysis-ready datasets, making it impossible to identify individual participants. Patients who were unwilling to participate or who withdrew their informed consent were excluded from the prospective registry.

[0047] Patient population

[0048] A total of 100 patients with stable chronic HFrEF enrolled in a prospective registry at the Medical University of Vienna Heart Failure Clinic, which was imputed with all clinically relevant data, were enrolled in this study. Enrollment criteria for the registry were documented chronic HFrEF, age at least 18 years, and receipt of stable heart failure-related medical therapy within the previous 3 months. Enrollment in the study was based on participation in the registry and the absence of any exclusion criteria preventing the prescribing of an SGLT-2 inhibitor (dapagliflozin). Clinic visits were conducted at baseline (Visit 0), 4 weeks after the start of SGLT-2 inhibitor therapy (Visit 1), 3 months later (Visit 2), 6 months later (Visit 3), and every 6 months thereafter.

[0049] Study endpoints: primary and secondary endpoints

[0050] All-cause mortality was defined as the primary endpoint. Predefined secondary endpoints included hospitalization for any cause and disease worsening, as assessed by worsening New York Heart Association (NYHA) functional class, change in NT-proBNP level, and change in left ventricular ejection fraction (LVEF). Associations between the primary and secondary endpoints were tested with respect to fluid status at baseline and its changes over time, glycemia (including blood glucose and glycated hemoglobin (HbA1c,%)), and renal function (creatinine and estimated glomerular filtration rate (eGFR)).

[0051] Demographics

[0052] Information on baseline characteristics, medical history, and current medications at enrollment and during follow-up will be recorded, including information on previous coronary angiography, right heart catheterization, echocardiographic data, myocardial death, myocardial infarction, hospitalization for heart failure, angina or implantation of an ICD and / or PM, and stroke.

[0053] Sampling and laboratory measurements

[0054] Before SGLT-2 inhibitor administration, blood and urine samples were collected from a peripheral vein at baseline and at each regularly scheduled study visit. The samples were used for (1.) routine laboratory analysis and (2.) biobank storage.

[0055] Biobank

[0056] Biobank blood was immediately centrifuged, and plasma samples were stored at −80° C. for future analysis. Plasma and urine samples were stored in the central “Biobundle” facility of the Department of Laboratory Medicine at the Medical University of Vienna.

[0057] Routine laboratory parameters

[0058] Routine laboratory parameters (such as blood cell count, CRP, creatinine, or GFR) and urinalysis were performed according to the standard procedures of the local laboratory.

[0059] Biomarker measurements

[0060] FABP3 and other myocardial biomarkers N-terminal B-type natriuretic peptide (NT-proBNP) and GDF-15 were measured in plasma using Elecsys Systems (Roche Diagnostics, Mannheim, Germany).

[0061] Bioelectrical impedance analysis

[0062] Bioelectrical impedance analysis (BIA) to assess fluid status was performed at enrollment before SGLT-2 inhibitor administration and at each scheduled study visit thereafter.

[0063] Echocardiographic evaluation

[0064] All patients underwent routine echocardiography using commercially available equipment (Vivid7, GE-Healthcare). Cardiac morphology was assessed using diameters in the four-chamber and two-chamber views, and ejection fraction was calculated using the biplane Simpson method according to EAE recommendations (Lang RM, Bierig M, Devereux RB, Flachskampf FA, Foster E, Pellikka PA et al. Recommendations for chamber quantification. European journal of echocardiography: the journal of the Working Group on Echocardiography of the European Society of Cardiology. 2006; 7(2): 79-108). Valvular morphology was assessed in standard sections according to guidelines and quantified by a comprehensive approach that included valve morphology and function (Zoghbi WA, Adams D, Bonow RO, Enriquez-Sarano M, Foster E, Grayburn PA et al. Recommendations for Noninvasive Evaluation of Native Valvular Regurgitation: A Report from the American Society of Echocardiography Developed in Collaboration with the Society for Cardiovascular Magnetic Resonance. Journal of the American Society of Echocardiography: official publication of the American Society of Echocardiography. 2017; 30(4): 303-71). Systolic pulmonary artery pressure (sPAP) was calculated as estimated central venous pressure. Raw data with a frame rate > 50 fps were digitally stored for post-processing.

[0065] Follow-up

[0066] Follow-up visits were scheduled as described above. In the event of a missed clinic visit, a telephone interview was performed. Blood and urine samples, and body fluid measurements were obtained at each visit. Echocardiography was repeated at Visit 3 (6 months after enrollment). Information on functional status, major cardiovascular events, and hospitalization for heart failure was recorded. In addition, mortality was ascertained by searching the Austrian Death Registry to minimize potential bias in the event of loss to follow-up.

[0067] result

[0068] Initiation of SGLT2i therapy is associated with increased plasma concentrations of FABP3 in patients susceptible and / or likely to respond to SGLT2i drugs ( Figure 1 ).like Figure 2 As shown in Figure 2, in the subgroup of patients with high baseline FABP3 (above the median concentration of 37.8 ng / mL), the amount of FABP3 at 3 months (V2) was approximately 14% higher than the amount of FABP3 at baseline. Figure 3 As shown, in the subgroup of patients with low baseline FABP3 (below the median concentration of 37.8 ng / mL), FABP3 levels at 3 months (V2) were approximately 27% higher than those at baseline. The FABP3 / NT-proBNP ratio increased over time after the initiation of SGLT2i, with the peak relative increase in FABP3 concentrations of 77% versus 19% (V3) occurring only after 6 months. Figure 4 In contrast, the amount of NT-proBNP before and after SGLT2i initiation was not associated with the patient's susceptibility and / or response to SGLT2i administration ( Figure 5 Furthermore, the FABP3 / GDF-15 ratio remained stable after SGLT2i initiation, with a peak relative increase of 12% at 12 months (V4) ( Figure 6 In this regard, the amount of GDF-15 did not change significantly before and after the start of SGLT2i ( Figure 7 ).

[0069] The present invention is also characterized by the following items.

[0070] 1. A method for detecting an effect induced by a sodium-glucose cotransporter inhibitor (SGLTi) in a subject suffering from cardiovascular disease, the method comprising a) determining the amount of fatty acid binding protein 3 (FABP3) in a sample from a subject treated with SGLTi; b) comparing the amount of FABP3 to a reference amount.

[0071] 2. The method according to item 1, wherein an increased amount of FABP3 compared to the reference amount is indicative of the effect.

[0072] 3. The method according to item 2, wherein an amount of FABP3 increased by at least 10% compared to the reference amount is indicative of the effect.

[0073] 4. The method according to any one of items 1 to 3, wherein the reference amount is the amount in a sample from a subject suffering from cardiovascular disease.

[0074] 5. The method according to any one of items 1 to 3, wherein the reference amount is the upper limit of normal (ULN) of the amount in a sample from a subject with cardiovascular disease.

[0075] 6. The method according to any one of items 1 to 3, wherein the reference amount is the amount in a sample from the same subject suffering from cardiovascular disease.

[0076] 7. The method according to any of the preceding items, wherein the reference amount is the amount in a sample from a subject with cardiovascular disease or the ULN of the amount in a sample from a subject with cardiovascular disease before treatment with SGLTi.

[0077] 8. The method according to any one of the preceding items, further comprising: c) determining the amount of FABP3 and cardiac injury markers in a sample of the subject treated with SGLTi, and comparing the amount of FABP3 and the amount of cardiac injury markers with reference amounts.

[0078] 9. The method according to any one of item 8, further comprising: d) generating a ratio between the amount of FABP3 and the amount of a cardiac damage marker and comparing the ratio with a reference ratio.

[0079] 10. The method according to item 9, wherein an increase in the ratio between the amount of FABP3 and the amount of a cardiac damage marker compared to the reference ratio is indicative of the effect.

[0080] 11. The method according to any one of the preceding claims, wherein the amount of FABP3 and optionally the amount of a cardiac injury marker is monitored, preferably wherein the amount of FABP3 and optionally the amount of a cardiac injury marker is monitored for at least 1 hour, at least 1 day, at least 1 week, more preferably at least 2 weeks or at least 4 weeks, more preferably 1 hour, 1 day, 7 days or 14 days.

[0081] 12. A method according to any one of items 8 to 11, wherein the cardiac injury marker comprises a cardiac function marker.

[0082] 13. The method according to any of the preceding items, wherein the cardiac marker is a natriuretic peptide, more preferably BNP or NT-proBNP, more preferably NT-proBNP.

[0083] 14. The method according to any one of items 8 to 13, wherein the cardiac damage marker is NT-proBNP, and the ratio between the amount of FABP3 and the amount of NT-proBNP is indicative of the effect, and the ratio is at least 0.030, preferably at least 0.032, more preferably at least 0.039.

[0084] 15. The method according to any one of the preceding items, wherein the subject suffering from cardiovascular disease suffers from heart failure, optionally chronic heart failure.

[0085] 16. The method according to any one of the preceding items, wherein the subject suffering from cardiovascular disease suffers from systolic or diastolic heart failure.

[0086] 17. The method according to any of the preceding items, wherein the subject with cardiovascular disease has an impaired left ventricular ejection fraction (LVEF).

[0087] 18. A method according to any of the preceding items, wherein the subject with cardiovascular disease suffers from heart failure classified as stage B, C or D according to the ACC / AHA classification, and / or wherein the subject with cardiovascular disease suffers from heart failure of class II to IV according to the NYHA classification.

[0088] 19. The method according to any one of the preceding items, wherein the sodium-glucose co-transporter inhibitor (SGLTi) comprises a sodium-glucose co-transporter 2 inhibitor (SGLT2i) or a sodium-glucose co-transporter 1 inhibitor (SGLT1i), preferably an SGLT2i, more preferably an SGLT2i, more preferably a gliflozin, more preferably comprising canagliflozin, dapagliflozin, empagliflozin, ertogliflozin, ipagliptin, repagliflozin, sergliflozin, soglipflozin or togliflozin.

[0089] 20. The method according to any one of the preceding items, wherein the subject is a human.

[0090] 21. The method according to any of the preceding items, wherein the sample is a blood, serum, plasma or urine sample, preferably the sample is a blood, serum or plasma sample.

[0091] 22. A computer-implemented method for performing the method of any of the preceding items, the computer-implemented method comprising: a) a means for determining the amount of FABP3 in a sample of a subject treated with SGLTi according to any of items 1 to 21 or determining the amount of FABP3 and a cardiac injury marker according to any of items 8 to 21, wherein the subject has cardiovascular disease; and b) a means for comparing the amount of FABP3 with a reference amount according to any of items 1 to 21, comparing the amount of FABP3 and the amount of the cardiac injury marker with a reference amount according to any of items 8 to 21, or generating a ratio between the amount of FABP3 and the amount of the cardiac injury marker according to any of items 9 to 21 and comparing the ratio with a reference ratio.

[0092] 23. A computer program product comprising program code for performing the computer-implemented method according to item 22 when run on at least one computer.

[0093] 24. A device for performing the method described in any of the preceding items, comprising: a) a means for determining the amount of FABP3 in a sample of a subject treated with SGLTi according to any of items 1 to 21 or determining the amount of FABP3 and a cardiac injury marker according to any of items 8 to 21, wherein the subject has cardiovascular disease; and b) a means for comparing the amount of FABP3 with a reference amount according to any of items 1 to 21, comparing the amount of FABP3 and the amount of the cardiac injury marker with a reference amount according to any of items 8 to 21, or generating a ratio between the amount of FABP3 and the amount of the cardiac injury marker according to any of items 9 to 21 and comparing the ratio with a reference ratio.

[0094] 25. A kit for performing the method of any one of the preceding items, comprising a) means for determining the amount of FABP3 in a sample of a subject treated with SGLTi according to any one of claims 1 to 21 or determining the amount of FABP3 and a cardiac marker according to any one of claims 8 to 21, wherein the subject suffers from cardiovascular disease; and b) means for comparing the amount of FABP3 with a reference amount according to any one of items 1 to 21, comparing the amount of FABP3 and the amount of the cardiac marker with a reference amount according to any one of items 8 to 21, or generating a ratio between the amount of FABP3 and the amount of the cardiac marker and comparing the ratio with a reference ratio according to any one of items 9 to 21.

[0095] 26. i) Use of FABP3 according to any one of items 1 to 21, FABP3 according to any one of items 8 to 21 and a cardiac damage marker and / or ii) use of a detection agent for FABP3 or for FABP3 and said cardiac marker for detecting an effect induced by a sodium-glucose cotransporter inhibitor (SGLTi) in a subject with cardiovascular disease.

[0096] It will be apparent to those skilled in the art that various substitutions and modifications can be made to the invention disclosed herein without departing from the scope and spirit of the invention.

[0097] All patents and publications mentioned in this specification are indicative of the levels of ordinary skill in the art to which this invention pertains. All patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.

[0098] The invention illustratively described herein may suitably be practiced in the absence of any one or more elements, restrictions or limitations not specifically disclosed herein. Thus, for example, the terms "comprising / including", "containing", etc. should be interpreted broadly and without limitation. Additionally, the terms and expressions employed herein have been used as descriptive and not restrictive terms, and when such terms and expressions are used, it is not intended to exclude any equivalents of the features shown and described or portions thereof, but it should be recognized that various modifications are possible within the scope of the claimed invention. Thus, it should be understood that although the invention has been specifically disclosed by preferred embodiments and optional features, modifications and variations of the invention disclosed herein may be made by those skilled in the art, and such modifications and variations are considered to be within the scope of the invention. The invention has been described broadly and generally herein. Each narrower species and subgeneric grouping falling within the general disclosure also forms part of the invention. This includes the general description of the invention with the proviso or negative limitation removing any subject matter from the genus, regardless of whether the excised material is specifically cited herein. In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group. Other embodiments of the invention will become apparent from the following claims.

Claims

1. A method for detecting an effect induced by a sodium-glucose cotransporter inhibitor (SGLTi) in a subject suffering from cardiovascular disease, the method comprising a) determining the amount of fatty acid binding protein 3 (FABP3) in a sample of said subject treated with SGLTi; b) comparing said amount of said FABP3 with a reference amount. 2 . The method of claim 1 , wherein an increased amount of FABP3 compared to the reference amount is indicative of the effect.

3. The method of claim 1 or 2, wherein the reference amount is the amount in a sample from a subject with cardiovascular disease, preferably wherein the reference amount is the amount in a sample from the same subject with cardiovascular disease; or wherein the reference amount is the upper limit of normal (ULN) for the amount in a sample from a subject with cardiovascular disease.

4. The method according to any one of the preceding claims, wherein the reference amount is the amount in a sample from a subject with cardiovascular disease before treatment with SGLTi or the ULN of the amount in a sample from a subject with cardiovascular disease.

5. The method according to any one of the preceding claims, further comprising: c) determining the amount of FABP3 and a cardiac injury marker in a sample from said subject treated with SGLTi, and comparing said amount of said FABP3 and said amount of said cardiac injury marker to a reference amount.

6. The method according to any one of claims 5, further comprising: d) generating a ratio between said amount of FABP3 and said amount of said cardiac injury marker and comparing said ratio to a reference ratio, preferably wherein an increased ratio between said amount of FABP3 and said amount of said cardiac injury marker compared to said reference ratio is indicative of said effect.

7. The method according to any one of the preceding claims, wherein the amount of FABP3 and optionally the amount of a cardiac injury marker is monitored, preferably wherein the amount of FABP3 and optionally the amount of a cardiac injury marker is monitored for at least 1 hour, at least 1 day, at least 1 week, more preferably at least 2 weeks, and even more preferably at least 4 weeks.

8. The method according to any one of claims 5 to 8, wherein the cardiac injury marker comprises a cardiac function marker, preferably wherein the cardiac injury marker is a natriuretic peptide, more preferably BNP or NT-proBNP, more preferably NT-proBNP.

9. The method according to any one of the preceding claims, wherein the subject suffering from cardiovascular disease suffers from heart failure, optionally chronic heart failure.

10. The method according to any one of the preceding claims, wherein the sodium-glucose co-transporter inhibitor (SGLTi) comprises a sodium-glucose co-transporter 2 inhibitor (SGLT2i) or a sodium-glucose co-transporter 1 inhibitor (SGLT1i), preferably an SGLT2i, more preferably an SGLT2i, more preferably a gliflozin, more preferably comprising canagliflozin, dapagliflozin, empagliflozin, ertogliflozin, ipagliptin, repagliflozin, sergliflozin, soglipflozin or togliflozin.

11. A computer-implemented method for performing the method according to any one of the preceding claims, the computer-implemented method comprising: a) means for determining the amount of FABP3 according to any one of claims 1 to 10 or the amount of FABP3 and the cardiac damage marker according to any one of claims 5 to 10 in a sample of a subject treated with SGLTi, wherein the subject suffers from cardiovascular disease; and b) means for comparing the amount of FABP3 with a reference amount according to any one of claims 1 to 10, for comparing the amount of FABP3 and the amount of the cardiac damage marker with a reference amount according to any one of claims 5 to 10, or for generating a ratio between the amount of FABP3 and the amount of the cardiac damage marker and comparing the ratio with a reference ratio according to any one of claims 6 to 10.

12. A computer program product comprising program code for performing the computer-implemented method according to claim 11 when run on at least one computer.

13. An apparatus for performing the method according to any one of the preceding claims, the apparatus comprising: a) means for determining the amount of FABP3 according to any one of claims 1 to 10 or the amount of FABP3 and the cardiac damage marker according to any one of claims 5 to 10 in a sample of a subject treated with SGLTi, wherein the subject suffers from cardiovascular disease; and b) means for comparing the amount of FABP3 with a reference amount according to any one of claims 1 to 10, for comparing the amount of FABP3 and the amount of the cardiac damage marker with a reference amount according to any one of claims 5 to 10, or for generating a ratio between the amount of FABP3 and the amount of the cardiac damage marker and comparing the ratio with a reference ratio according to any one of claims 6 to 10.

14. A kit for performing the method according to any one of the preceding claims, comprising: a) means for determining the amount of FABP3 according to any one of claims 1 to 10 or the amount of FABP3 and a cardiac marker according to any one of claims 5 to 10 in a sample of a subject treated with SGLTi, wherein the subject suffers from a cardiovascular disease; and b) means for comparing the amount of FABP3 with a reference amount according to any one of claims 1 to 10, for comparing the amount of FABP3 and the amount of the cardiac marker with a reference amount according to any one of claims 5 to 10, or for generating a ratio between the amount of FABP3 and the amount of the cardiac marker and comparing the ratio with a reference ratio according to any one of claims 6 to 10.

15. i) Use of FABP3 according to any one of claims 1 to 10, FABP3 according to any one of claims 5 to 10 and cardiac damage markers and / or ii) Use of a detection agent for FABP3 or for FABP3 and cardiac markers for detecting effects induced by sodium-glucose cotransporter inhibitors (SGLTi) in subjects with cardiovascular disease.

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