WO2016007945A1 - Compositions and methods for treating or preventing cardiac hypertrophy - Google Patents

Abstract

The invention relates generally to compositions and methods featuring for treating or preventing a cardiac condition induced by phosphoinositide 3-kinase (PI3K) inhibitor treatment.

Description

COMPOSITIONS AND METHODS FOR TREATING OR PREVENTING

CARDIAC HYPERTROPHY

RELATED APPLICATIONS

This application claims the benefit of priority under 35 U.S.C. § 119(e) to U.S.

Provisional Application No: 62/023,245, filed July 11, 2014, which is incorporated herein by reference in its entirety.

BACKGROUND

Cardiac hypertrophy relates to the thickening of the heart muscle (myocardium), which results in a decrease in size of the chamber of the heart, including the left and right ventricles. The heart becomes weaker and is not as efficient in pumping and circulating blood. As such, there is a pressing need to develop new strategies to treat cardiac

hypertrophy.

SUMMARY OF THE INVENTION

The invention provides methods of treating phosphoinositide 3 -kinase (PI3K) inhibitor-induced cardiotoxicity by administering an effective amount of an insulin-like growth factor 1 receptor (IGFR)/insulin receptor (IR) inhibitor to a subject in need thereof, thereby treating the PI3K inhibitor- induced cardiotoxicity. In some cases, the subject is identified by echocardiography. The subject is preferably a mammal in need of such treatment, e.g., a subject that has been diagnosed with cardiotoxicity or a predisposition thereto. The mammal is any mammal, e.g., a human, a primate, a mouse, a rat, a dog, a cat, a horse, as well as livestock or animals grown for food consumption, e.g., cattle, sheep, pigs, chickens, and goats. In a preferred embodiment, the mammal is a human. Preferably, cardiac hypertrophy and adverse cardiac remodeling is reduced in the subject following

administration.

Preferably, the effective amount of IGFR/IR inhibitor is sufficient to reduce cardiac hypertrophy by at least 5%, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or by 100%. In some cases, the effective amount of IGFR/IR inhibitor is sufficient to reduce adverse cardiac remodeling by at least 5%, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or by 100%. An exemplary IGFR/IR inhibitor comprises 3-[8-Amino-l-(2-phenyl-7- quinolyl)imidazo[l,5-a]pyrazin-3-yl]-l-methyl-cyclobutanol (OSI-906). The effective amount of OSI-906 is from 0.5 mg/kg to 100 mg/kg, e.g., 1 mg/kg, 5 mg/kg, 10 mg/kg, 25 mg/kg, 50 mg/kg, 75 mg/kg, or 100 mg/kg body weight. In one aspect, OSI-906 is administered at a low dose, e.g., 0.5 mg/kg- 1 mg/kg body weight. The OSI-906 is administered at least once per hour, e.g., at least once per day, at least once per week, or at least once per month. In some cases, the IGFR IR inhibitor is administered daily, e.g., every 24 hours. Or, the IGFR/IR inhibitor is administered continuously or several times per day, e.g., every 1 hour, every 2 hours, every 3 hours, every 4 hours, every 5 hours, every 6 hours, every 7 hours, every 8 hours, every 9 hours, every 10 hours, every 11 hours, or every 12 hours. Suitable modes of administration include oral, intramuscular, intravenous, subcutaneous, and systemic. The OSI-906 is administered prior to, concurrently with, or after administration of the PI3K inhibitor.

Ultimately, the attending physician or veterinarian decides the appropriate amount and dosage regimen.

Suitable PI3K inhibitors are selected from the group consisting of wortmannin, demethoxyviridin, LY294002, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, GDC-0941, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477, CUDC-907, BYL719, GDC- 0032, BGT226, GDC0980, PF4691502, PKI587, and AEZS-136.4. An exemplary PI3K inhibitor for use in the methods described herein comprises 2-methyl-2(4-[3-methyl-2-oxo-8- (quinolin-3-yl)-2,3-dihydro-lH-imidazo[4,5-c]quinolin-l-yl]phenyl)propanenitrile (BEZ- 235).

Also provided are methods of enhancing cardiac function or increasing survival in a subject treated with a PI3K inhibitor comprising administering an effective amount of OSI- 906 to the subject, thereby improving cardiac function in the subject. For example, cardiac function is enhanced by at least 5%, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or by 100%.

Kits for preventing or treating PI3K inhibitor-induced cardiotoxicity comprise an effective amount of OSI-906 and instructions for use. The invention also provides a pharmaceutical composition comprising an effective amount of BEZ-235 and an effective amount of OSI-906, and instructions for the

administration of each.

Definitions

By "OSI-906" or "3-[8-Arnino-l-(2-phenyl-7-quinolyl)imidazo[l,5-a]pyrazin-3-yl]-l- methyl-cyclobutanol" is meant an IGFR/IR inhibitor having the following structure:

By "BEZ235" or "2-methyl-2(4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydro-lH- imidazo[4,5-c]quinolin-l-yl]phenyl)propanenitrile" is meant a PI3K inhibitor having the followin structure:

By "PI3K inhibitor" is meant an agent that inhibits the activity of phosphoinositide 3- kinase. Non- limiting examples of PI3K inhibitors include wortmannin, demethoxyviridin, LY294002, perifosine, CALlOl, PX-866, BEZ235, SF1126, INK1117, IPI-145, GDC-0941, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

By "cardiac hypertrophy" is meant any undesirable cardiac muscle growth, increase in cardiac chamber mass relative to body size, or increase in cardiac chamber wall thickness at normal or increased chamber volume.

By "enhancing cardiac function" is meant producing a beneficial alteration in the pumping performance and capacity of the heart. In one embodiment, the method increases cardiac function by at least about 10%, 25%, 50%, 75% or more. Methods for measuring cardiac function are known in the art and described herein.

By "echocardiography" or "cardiac ultrasound" is meant a sonogram of the heart used to generate an accurate assessment of the velocity of blood at any point during the cardiac cycle.

By "end-systolic volume (ESV)" is meant the volume of blood in a ventricle at the end of contraction and represents the smallest volume of blood in the ventricle at any point in the cardiac cycle. By "end-diastolic volume (EDV)" is meant the volume of blood in a ventricle at the end load or filing in.

By "increasing survival" is meant an increase in the amount of time a treated subject lives relative to an untreated corresponding control subject. For example, an increase in survival of at least about 1, 2, 3, 4, or 5 weeks. Preferably, survival is increased by at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In other embodiments, survival is increased by at least 1, 2, 3, 4, or 5 years.

By "agent" is meant any small molecule chemical compound, antibody, nucleic acid molecule, or polypeptide, or fragments thereof.

A small molecule is a compound that is less than 2000 Daltons in mass. The molecular mass of the small molecule is preferably less than 1000 Daltons, more preferably less than 600 Daltons, e.g., the compound is less than 500 Daltons, less than 400 Daltons, less than 300 Daltons, less than 200 Daltons, or less than 100 Daltons.

Small molecules are organic or inorganic. Exemplary organic small molecules include, but are not limited to, aliphatic hydrocarbons, alcohols, aldehydes, ketones, organic acids, esters, mono- and disaccharides, aromatic hydrocarbons, amino acids, and lipids.

Exemplary inorganic small molecules comprise trace minerals, ions, free radicals, and metabolites. Alternatively, small molecules can be synthetically engineered to consist of a fragment, or small portion, or a longer amino acid chain to fill a binding pocket of an enzyme. Typically small molecules are less than one kilodalton.

In some cases, a compound (e.g., small molecule) or macromolecule (e.g., nucleic acid, polypeptide, or protein) of the invention is purified and/or isolated. As used herein, an "isolated" or "purified" small molecule, nucleic acid molecule, polynucleotide, polypeptide, or protein (e.g., antibody or fragment thereof), is substantially free of other cellular material, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized. Purified compounds are at least 60% by weight (dry weight) the compound of interest. Preferably, the preparation is at least 75%, more preferably at least 90%, and most preferably at least 99%, by weight the compound of interest. For example, a purified compound is one that is at least 90%, 91%, 92%, 93%, 94%, 95%, 98%, 99%, or 100% (w/w) of the desired compound by weight. Purity is measured by any appropriate standard method, for example, by column chromatography, thin layer chromatography, or high-performance liquid chromatography (HPLC) analysis. A purified or isolated polynucleotide (ribonucleic acid (RNA) or deoxyribonucleic acid (DNA), e.g., synthetic cDNA) is free of the genes or sequences that flank it in its naturally occurring state. Purified also defines a degree of sterility that is safe for administration to a human subject, e.g. , lacking infectious or toxic agents.

By "ameliorate" is meant decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a disease.

By "alteration" is meant a change (increase or decrease) in a clinical parameter or biomarker (e.g., polypeptide, polynucleotide level) as detected by standard art known methods such as those described herein. As used herein, an alteration includes a 10%, 25%, 40%, 50% or greater change.

By "disease" is meant any condition or disorder that damages or interferes with the normal function of a cell, tissue, or organ.

By "modulation" is meant any alteration (e.g., increase or decrease) in a biological function or activity.

As used herein, "obtaining" as in "obtaining an agent" includes synthesizing, purchasing, or otherwise acquiring the agent.

By "increases" or reduces" is meant a positive or negative alteration, respectively, of at least about 10%, 25%, 50%, 75%, or 100%.

By "control" or "reference" is meant a standard of comparison. As used herein, "changed as compared to a control" sample or subject is understood as having a level of the analyte or diagnostic or therapeutic indicator to be detected at a level that is statistically different than a sample from a normal, untreated, or control sample. Control samples include, for example, cells in culture, one or more laboratory test animals, or one or more human subjects. Methods to select and test control samples are within the ability of those in the art. An analyte can be a naturally occurring substance that is characteristically expressed or produced by the cell or organism (e.g., an antibody, a protein) or a substance produced by a reporter construct (e.g, β-galactosidase or luciferase). Depending on the method used for detection, the amount and measurement of the change can vary. Determination of statistical significance is within the ability of those skilled in the art, e.g., the number of standard deviations from the mean that constitute a positive result.

As used herein, "detecting" and "detection" are understood that an assay performed for identification of a specific analyte in a sample, e.g., an antigen in a sample or the level of an antigen in a sample. The amount of analyte or activity detected in the sample can be none or below the level of detection of the assay or method.

By "diagnosing" as used herein refers to a clinical or other assessment of the condition of a subject based on observation, testing, or circumstances for identifying a subject having a disease, disorder, or condition (e.g., cardiotoxicity) based on the presence of at least one indicator, such as a sign or symptom of the disease, disorder, or condition. Typically, diagnosing using the method of the invention includes the observation of the subject for multiple indicators of the disease, disorder, or condition in conjunction with the methods provided herein. A diagnostic method provides an indicator that a disease is or is not present.

A single diagnostic test typically does not provide a definitive conclusion regarding the disease state of the subject being tested.

By "specifically binds" is meant a compound or antibody that recognizes and binds a polypeptide of the invention, but which does not substantially recognize and bind other molecules in a sample, for example, a biological sample, which naturally includes a polypeptide of the invention.

Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,

41, 42, 43, 44, 45, 46, 47, 48, 49, or 50.

By the terms "effective amount" and "therapeutically effective amount" of a formulation or formulation component is meant a sufficient amount of the formulation or component, alone or in a combination, to provide the desired effect. For example, by "an effective amount" is meant an amount of a compound, alone or in a combination, required to reduce or prevent cardiotoxicity in a mammal. Ultimately, the attending physician or veterinarian decides the appropriate amount and dosage regimen.

"Cardiotoxicity" is the occurrence of heart electrophysiology dysfunction or muscle damage. The heart becomes weaker and is not as efficient in pumping and therefore circulating blood. Cardiotoxicity may be caused by chemotherapy treatment, complications from anorexia nervosa, adverse effects of heavy metals intake, or an incorrectly administered drug such as bupivacaine. Thus, PI3K inhibitor-induced cardiotoxicity comprises cardiotoxicity induced by a PI3K inhibitor.

The terms "treating" and "treatment" as used herein refer to the administration of an agent or formulation to a clinically symptomatic individual afflicted with an adverse condition, disorder, or disease, so as to effect a reduction in severity and/or frequency of symptoms, eliminate the symptoms and/or their underlying cause, and/or facilitate improvement or remediation of damage. The terms "preventing" and "prevention" refer to the administration of an agent or composition to a clinically asymptomatic individual who is susceptible or predisposed to a particular adverse condition, disorder, or disease, and thus relates to the prevention of the occurrence of symptoms and/or their underlying cause.

Ranges provided herein are understood to be shorthand for all of the values within the range. This includes all individual sequences when a range of SEQ ID NOs: is provided. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50.

Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive.

Unless specifically stated or obvious from context, as used herein, the terms "a", "an", and "the" are understood to be singular or plural.

Unless specifically stated or obvious from context, as used herein, the term "about" is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term about.

The transitional term "comprising," which is synonymous with "including,"

"containing," or "characterized by," is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. By contrast, the transitional phrase

"consisting of excludes any element, step, or ingredient not specified in the claim. The transitional phrase "consisting essentially of limits the scope of a claim to the specified materials or steps "and those that do not materially affect the basic and novel

characteristic(s)" of the claimed invention.

The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein.

Other features and advantages of the invention will be apparent from the following description of the preferred embodiments thereof, and from the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All published foreign patents and patent applications cited herein are incorporated herein by reference. Genbank and NCBI submissions indicated by accession number cited herein are incorporated herein by reference. All other published references, documents, manuscripts and scientific literature cited herein are incorporated herein by reference. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 A-Figure IF is a series of bar charts and a photomicrograph of an immunoblot showing BEZ induced cardiac hypertrophy in mice. Three months old FVB/n female mice (n=10-15 per group) were treated with BEZ (15, 50, 100 mg/kg, daily, five days a week, oral gavage). 5-6 weeks after the treatment the following studies were performed: (Figure 1A) Echocardiography (EDV: end-diastolic volume; ESV: end-systolic volume; SWth: septum wall thickness; PWth: posterior wall thickness: EF%: ejection fraction; RWth: relative wall thickness); (Figure IB) Hemodynamic measurements (HR: heart rate; LVSP: left ventricular end-systolic pressure; LVW/TL: ratio of LV weight to tibia length; dP/dt: the rate of LV pressure rise or decline); (Figure 1C) Morphology studies for cardiomyocyte cross sectional area and cardiac fibrosis; (Figure ID) Real-time PCR analysis of cardiac hypertrophic marker fetal gene RNA expressions; (Figure 1E-F) Western blot analysis of phosphorylation (activation) of the insulin receptor proteins and downstream signaling pathways.

Figure 2A-Figure 2H is a series of line graphs, bar charts, and an immunoblot showing that BEZ increased hepatic gluconeogenesis, induced hyperglycemia,

hyperinsulinemia, and increased insulin receptor activation in the heart. Figure 2A-Figure 2B: Fasting glucose: Mice (n=5-8 per point) were treated with BEZ, and fasted. Fasting glucose was measured 8 hours after the treatment. Figure 2C: Glucose tolerance test: Mice (n=5-9 per group) were fasted overnight; GTT was performed 16-18 hours after fasting. Figure 2D: Pyruvate tolerance test: Mice (n=5-8 per group) were fasted overnight; PTT was performed 16-18 hours after fasting. Figure 2E: Real-time PCR of hepatic gluconeogenesis gene expression: Mice (n=5 per group) were treated with BEZ and then fasted and sacrificed at different time points. mRNA expression of phosphoenolpyruvate carboxykinase (PCK1) and glucose-6-phosphatase (G6PC) in liver were measured by real-time RT-PCR. Figure 2F: Serum insulin: Mice (n=5 per point per group) were treated with BEZ and then fasted and sacrificed at different time points. Serum insulin was measured by ELISA. Figure 2G- Figure 2H: Western blot analysis of IR and downstream signaling pathways: Mice (n=5 per point, per group) were treated with BEZ and then fasted and sacrificed at different time points. Activation of the insulin receptor (IR) and its downstream signaling pathways (PI3K- Akt, mTOR) were assessed by Western blotting using specific antibodies.

Figure 3A-Figure 3G is a series of bar charts, line graphs, and a schematic demonstrating that injections of insulin worsened, whereas blocking IGFR/IR by OSI- 906 alleviated BEZ-induced cardiac hypertrophy. Mice (n=5-10 per group) were treated with BEZ with or without concurrent administration of insulin (0.5, 2, 5, 10, 20 IU, i.p. daily, five days a week) or OSI- 906 (0.5 mg/kg, oral gavage, daily, five days a week).

Echocardiography and hemodynamic measurements were performed 4-5 weeks after the treatments. Figure 3A: Echocardiography. Figure 3B: Hemodynamic measurements. Figure 3C: Fasting glucose, GTT and PTT. Figure 3D: Insulin receptor activation in the heart. Figure 3E: Echocardiography in OSI treated mice. Figure 3F: Hemodynamic measurements in OSI treated mice. Figure 3G is a schematic illustrating the ability of OSI-906 to prevent cardiomyocyte growth.

DETAILED DESCRIPTION OF THE INVENTION

As described in detail below, a dual PI3K-mTOR inhibitor, BEZ235 (BEZ), induced cardiac hypertrophy, hyperglycemia, and increased insulin signals in the heart. Three month old FVB/n female mice were treated with BEZ for five weeks. Cardiac function was monitored by serial echocardiography during the treatment and hemodynamic measurements at the end of the study. Cell signaling was analyzed by RT-PCR, Western blotting, and ELISA. BEZ induced a dose-dependent increase of left ventricular (LV) wall thickness and systolic function. These were associated with increased hypertrophic markers ANP, BNP, β- MHC and a-skeletal actin in the heart. In addition, in chronic BEZ-treated mouse hearts, the activations of PI3Ks, mTOR and ERK were increased. Further studies were conducted to understand these contradictory results. As described below, BEZ induced an increase of hepatic gluconeogenesis gene expression, which was associated with increased fasting glucose, increased serum insulin level, a worsened glucose and pyruvate tolerance and increased IGFR/Insulin receptor activation in the heart. Injections of insulin lowered blood glucose, improved glucose and pyruvate tolerance, but further aggravated BEZ-induced cardiac dysfunction. On the other hand, OSI-906 (an IGFR/IR inhibitor) normalized cardiac function in BEZ-treated mice. These results indicate that chronic BEZ treatment induced cardiac hypertrophy is caused by increased insulin receptor activation in the heart.

Any number of standard methods are available for assaying cardiovascular function.

Preferably, cardiovascular function in a subject (e.g., a human) is assessed using non-invasive means, such as measuring net cardiac ejection (ejection fraction, fractional shortening, and ventricular end-systolic volume) by an imaging method such as echocardiography, nuclear or radiocontrast ventriculography, or magnetic resonance imaging, and systolic tissue velocity as measured by tissue Doppler imaging. Systolic contractility can also be measured non- invasively using blood pressure measurements combined with assessment of heart outflow (to assess power), or with volumes (to assess peak muscle stiffening). Measures of

cardiovascular diastolic function include ventricular compliance, which is typically measured by the simultaneous measurement of pressure and volume, early diastolic left ventricular filling rate and relaxation rate (can be assessed from echoDoppler measurements). Other measures of cardiac function include myocardial contractility, resting stroke volume, resting heart rate, resting cardiac index (cardiac output per unit of time [L/minute], measured while seated and divided by body surface area [m²])) total aerobic capacity, cardiovascular performance during exercise, peak exercise capacity, peak oxygen (0₂) consumption, or by any other method known in the art or described herein. Measures of vascular function include determination of total ventricular afterload, which depends on a number of factors, including peripheral vascular resistance, aortic impedance, arterial compliance, wave reflections, and aortic pulse wave velocity.

Methods for assaying cardiovascular function include any one or more of the following: Doppler echocardiography, 2-dimensional echo-Doppler imaging, pulse-wave Doppler, continuous wave Doppler, oscillometric arm cuff, tissue Doppler imaging, cardiac catheterization, magnetic resonance imaging, positron emission tomography, chest X-ray, X- ray contrast ventriculography, nuclear imaging ventriculography, computed tomography imaging, rapid spiral computerized tomographic imaging, 3-D echocardiography, invasive cardiac pressures, invasive cardiac flows, invasive cardiac pressure- volume loops

(conductance catheter), non-invasive cardiac pressure-volume loops.

The therapeutic methods of the invention (which include prophylactic treatment) in general comprise administration of a therapeutically effective amount of the compounds herein, such as a compound of the formulae herein to a subject (e.g., animal, human) in need thereof, including a mammal, particularly a human. Such treatment will be suitably administered to subjects, particularly humans, suffering from, having, susceptible to, or at risk for a disease, disorder, or symptom thereof. Determination of those subjects "at risk" can be made by any objective or subjective determination by a diagnostic test or opinion of a subject or health care provider (e.g., genetic test, enzyme or protein marker, Marker (as defined herein), family history, and the like).

Pharmaceutical Compositions

The present invention features pharmaceutical preparations comprising OSI-906 together with pharmaceutically acceptable carriers, where the compounds provide for the treatment of virtually any cardiac indication induced by a PI3K inhibitor. Pharmaceutical preparations of the invention have both therapeutic and prophylactic applications. In one embodiment, a pharmaceutical composition includes an effective amount of OSI-906. The compositions should be sterile and contain a therapeutically effective amount of OSI-906 in a unit of weight or volume suitable for administration to a subject (e.g., a human patient). The compositions and combinations of the invention can be part of a pharmaceutical pack, where the OSI-906 is present in individual dosage amounts.

Pharmaceutical compositions of the invention to be used for prophylactic or therapeutic administration should be sterile. Sterility is readily accomplished by filtration through sterile filtration membranes (e.g., 0.2 μιη membranes), by gamma irradiation, or any other suitable means known to those skilled in the art. Therapeutic compositions generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle. These

compositions ordinarily will be stored in unit or multi-dose containers, for example, sealed ampoules or vials, as an aqueous solution or as a lyophilized formulation for reconstitution.

OSI-906 may be combined, optionally, with a pharmaceutically acceptable excipient.

The term "pharmaceutically-acceptable excipient" as used herein means one or more compatible solid or liquid filler, diluents or encapsulating substances that are suitable for administration into a human. The term "carrier" denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate administration. The components of the pharmaceutical compositions also are capable of being co-mingled with OSI-906, and with each other, in a manner such that there is no interaction that would substantially impair the desired pharmaceutical efficacy.

Compounds of the present invention can be contained in a pharmaceutically acceptable excipient. The excipient preferably contains minor amounts of additives such as substances that enhance isotonicity and chemical stability. Such materials are non-toxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, succinate, acetate, lactate, tartrate, and other organic acids or their salts; tris-hydroxymethylaminomethane (TRIS), bicarbonate, carbonate, and other organic bases and their salts; antioxidants, such as ascorbic acid; low molecular weight (for example, less than about ten residues) polypeptides, e.g., polyarginine, polylysine, polyglutamate and polyaspartate; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone (PVP), polypropylene glycols (PPGs), and polyethylene glycols (PEGs); amino acids, such as glycine, glutamic acid, aspartic acid, histidine, lysine, or arginine; monosaccharides, disaccharides, and other carbohydrates including cellulose or its derivatives, glucose, mannose, sucrose, dextrins or sulfated carbohydrate derivatives, such as heparin, chondroitin sulfate or dextran sulfate; polyvalent metal ions, such as divalent metal ions including calcium ions, magnesium ions and manganese ions; chelating agents, such as ethylenediamine tetraacetic acid (EDTA); sugar alcohols, such as mannitol or sorbitol; counterions, such as sodium or ammonium; and/or nonionic surfactants, such as polysorbates or poloxamers. Other additives may be included, such as stabilizers, anti-microbials, inert gases, fluid and nutrient replenishers (i.e., Ringer's dextrose), electrolyte replenishers, and the like, which can be present in conventional amounts.

The compositions, as described above, can be administered in effective amounts. The effective amount will depend upon the mode of administration, the particular condition being treated and the desired outcome. It may also depend upon the stage of the condition, the age and physical condition of the subject, the nature of concurrent therapy, if any, and like factors well known to the medical practitioner. For therapeutic applications, it is that amount sufficient to achieve a medically desirable result.

With respect to a subject having a cardiac disease or disorder induced by a PI3K inhibitor, an effective amount is sufficient to prevent, reduce, stabilize, or reverse an alteration associated with cardiotoxicity induced by the PI3K inhibitor. With respect to a subject having a cardiac disease or disorder induced by PI3K inhibitor, an effective amount is an amount sufficient to stabilize, slow, or reduce a symptom associated with the cardiac condition. Generally, doses of the compounds of the present invention would be from about 0.01 mg/kg per day to about 1000 mg/kg per day. In one embodiment, 25, 50, 75, 100, 125, 150 or 200 mg/kg bodyweight of OSI-906 is administered to a subject. Preferably, 25 to 100 mg/kg of OSI-906 is administered. Desirably, the OSI-906 is administered in an amount sufficient to achieve a peak concentration in plasma. It is expected that doses ranging from about 5 to about 2000 mg/kg will be suitable. Lower doses will result from certain forms of administration, such as intravenous administration and pharmaceutical. In the event that a response in a subject is insufficient at the initial doses applied, higher doses (or effectively higher doses by a different, more localized delivery route) may be employed to the extent that patient tolerance permits. Multiple doses per day are contemplated to achieve appropriate systemic levels of a composition of the present invention.

A variety of administration routes are available. The methods of the invention, generally speaking, may be practiced using any mode of administration that is medically acceptable, meaning any mode that produces effective levels of the active compounds without causing clinically unacceptable adverse effects. In one preferred embodiment, a composition of the invention is administered orally. Other modes of administration include rectal, topical, intraocular, buccal, intravaginal, intracisternal, intracerebroventricular, intratracheal, nasal, transdermal, within/on implants, or parenteral routes. The term

"parenteral" includes subcutaneous, intrathecal, intravenous, intramuscular, intraperitoneal, or infusion. Intravenous or intramuscular routes are not particularly suitable for long-term therapy and prophylaxis. They could, however, be preferred in emergency situations.

Compositions comprising a composition of the invention can be added to a physiological fluid, such as blood. Oral administration can be preferred for prophylactic treatment because of the convenience to the patient as well as the dosing schedule.

Pharmaceutical compositions of the invention can comprise one or more pH buffering compounds to maintain the pH of the formulation at a predetermined level that reflects physiological pH, such as in the range of about 5.0 to about 8.0. The pH buffering compound used in the aqueous liquid formulation can be an amino acid or mixture of amino acids, such as histidine or a mixture of amino acids such as histidine and glycine. Alternatively, the pH buffering compound is preferably an agent which maintains the pH of the formulation at a predetermined level, such as in the range of about 5.0 to about 8.0, and which does not chelate calcium ions. Illustrative examples of such pH buffering compounds include, but are not limited to, imidazole and acetate ions. The pH buffering compound may be present in any amount suitable to maintain the pH of the formulation at a predetermined level.

Pharmaceutical compositions of the invention can also contain one or more osmotic modulating agents, i.e., a compound that modulates the osmotic properties (e.g., tonicity, osmolality and/or osmotic pressure) of the formulation to a level that is acceptable to the blood stream and blood cells of recipient individuals. The osmotic modulating agent can be an agent that does not chelate calcium ions. The osmotic modulating agent can be any compound known or available to those skilled in the art that modulates the osmotic properties of the formulation. One skilled in the art may empirically determine the suitability of a given osmotic modulating agent for use in the inventive formulation. Illustrative examples of suitable types of osmotic modulating agents include, but are not limited to: salts, such as sodium chloride and sodium acetate; sugars, such as sucrose, dextrose, and mannitol; amino acids, such as glycine; and mixtures of one or more of these agents and/or types of agents. The osmotic modulating agent(s) may be present in any concentration sufficient to modulate the osmotic properties of the formulation.

Compositions comprising a compound of the present invention can contain multivalent metal ions, such as calcium ions, magnesium ions and/or manganese ions. Any multivalent metal ion that helps stabilizes the composition and that will not adversely affect recipient individuals may be used. The skilled artisan, based on these two criteria, can determine suitable metal ions empirically and suitable sources of such metal ions are known, and include inorganic and organic salts.

Pharmaceutical compositions of the invention can also be a non-aqueous liquid formulation. Any suitable non-aqueous liquid may be employed, provided that it provides stability to the active agents (s) contained therein. Preferably, the non-aqueous liquid is a hydrophilic liquid. Illustrative examples of suitable non-aqueous liquids include: glycerol; dimethyl sulfoxide (DMSO); polydimethylsiloxane (PMS); ethylene glycols, such as ethylene glycol, diethylene glycol, Methylene glycol, polyethylene glycol ("PEG") 200, PEG 300, and PEG 400; and propylene glycols, such as dipropylene glycol, tripropylene glycol, polypropylene glycol ("PPG") 425, PPG 725, PPG 1000, PPG 2000, PPG 3000 and PPG 4000.

Pharmaceutical compositions of the invention can also be a mixed aqueous/non- aqueous liquid formulation. Any suitable non-aqueous liquid formulation, such as those described above, can be employed along with any aqueous liquid formulation, such as those described above, provided that the mixed aqueous/non- aqueous liquid formulation provides stability to the compound contained therein. Preferably, the non-aqueous liquid in such a formulation is a hydrophilic liquid. Illustrative examples of suitable non-aqueous liquids include: glycerol; DMSO; PMS; ethylene glycols, such as PEG 200, PEG 300, and PEG 400; and propylene glycols, such as PPG 425, PPG 725, PPG 1000, PPG 2000, PPG 3000 and PPG 4000. Suitable stable formulations can permit storage of the active agents in a frozen or an unfrozen liquid state. Stable liquid formulations can be stored at a temperature of at least - 70°C, but can also be stored at higher temperatures of at least 0°C, or between about 0.1 °C. and about 42°C, depending on the properties of the composition. It is generally known to the skilled artisan that proteins and polypeptides are sensitive to changes in pH, temperature, and a multiplicity of other factors that may affect therapeutic efficacy.

Other delivery systems can include time-release, delayed release or sustained release delivery systems. Such systems can avoid repeated administrations of compositions of the invention, increasing convenience to the subject and the physician. Many types of release delivery systems are available and known to those of ordinary skill in the art. They include polymer base systems such as polylactides (U.S. Pat. No. 3,773,919; European Patent No. 58,481), poly(lactide-glycolide), copolyoxalates, polycaprolactones, polyesteramides, polyorthoesters, polyhydroxybutyric acids, such as poly-D-(-)-3-hydroxybutyric acid (European Patent No. 133, 988), copolymers of L-glutamic acid and gamma-ethyl-L- glutamate (Sidman, K. R. et al., Biopolymers 22: 547-556), poly (2-hydroxyethyl methacrylate) or ethylene vinyl acetate (Langer, R. et al., J. Biomed. Mater. Res. 15:267-277; Langer, R. Chem. Tech. 12:98-105), and poly anhydrides.

Other examples of sustained-release compositions include semi-permeable polymer matrices in the form of shaped articles, e.g., films, or microcapsules. Delivery systems also include non-polymer systems that are: lipids including sterols such as cholesterol, cholesterol esters and fatty acids or neutral fats such as mono- di- and tri-glycerides; hydrogel release systems such as biologically-derived bioresorbable hydrogel (i.e., chitin hydrogels or chitosan hydrogels); sylastic systems; peptide based systems; wax coatings; compressed tablets using conventional binders and excipients; partially fused implants; and the like. Specific examples include, but are not limited to: (a) erosional systems in which the agent is contained in a form within a matrix such as those described in U.S. Pat. Nos. 4,452,775, 4,667,014, 4,748,034 and 5,239,660 and (b) diffusional systems in which an active component permeates at a controlled rate from a polymer such as described in U.S. Pat. Nos. 3,832,253, and 3,854,480.

Another type of delivery system that can be used with the methods and compositions of the invention is a colloidal dispersion system. Colloidal dispersion systems include lipid- based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. Liposomes are artificial membrane vessels, which are useful as a delivery vector in vivo or in vitro. Large unilamellar vessels (LUV), which range in size from 0.2-4.0 μιη, can encapsulate large macromolecules within the aqueous interior and be delivered to cells in a biologically active form (Fraley, R., and Papahadjopoulos, D., Trends Biochem. Sci. 6: 77- 80).

Liposomes can be targeted to a particular tissue by coupling the liposome to a specific ligand such as a monoclonal antibody, sugar, glycolipid, or protein. Liposomes are commercially available from Gibco BRL, for example, as LIPOFECTIN.TM. and

LIPOFECTACE.TM., which are formed of cationic lipids such as N-[l-(2,3 dioleyloxy)- propyl]-N,N,N-trimethylammonium chloride (DOTMA) and dimethyl dioctadecylammonium bromide (DDAB). Methods for making liposomes are well known in the art and have been described in many publications, for example, in DE 3,218,121; Epstein et al., Proc. Natl.

Acad. Sci. (USA) 82:3688-3692 (1985); Hwang et al., Proc. Natl. Acad. Sci. (USA) 77:4030- 4034 (1980); EP 52,322; EP 36,676; EP 88,046; EP 143,949; EP 142,641; Japanese Pat. Appl. 83-118008; U.S. Pat. Nos. 4,485,045 and 4,544,545; and EP 102,324. Liposomes also have been reviewed by Gregoriadis, G., Trends Biotechnol., 3: 235-241).

Another type of vehicle is a biocompatible microparticle or implant that is suitable for implantation into a mammalian recipient. Exemplary bioerodible implants that are useful in accordance with this method are described in PCT International application no.

PCT/US/03307 (Publication No. WO 95/24929, entitled "Polymeric Gene Delivery System"). PCT/US/0307 describes biocompatible, preferably biodegradable polymeric matrices for containing an exogenous gene under the control of an appropriate promoter. The polymeric matrices can be used to achieve sustained release of the exogenous gene or gene product in the subject.

The polymeric matrix preferably is in the form of a microparticle such as a microsphere (wherein an agent is dispersed throughout a solid polymeric matrix) or a microcapsule (wherein an agent is stored in the core of a polymeric shell). Microcapsules of the foregoing polymers containing drugs are described in, for example, U.S. Pat. No.

5,075,109. Other forms of the polymeric matrix for containing an agent include films, coatings, gels, implants, and stents. The size and composition of the polymeric matrix device is selected to result in favorable release kinetics in the tissue into which the matrix is introduced. The size of the polymeric matrix further is selected according to the method of delivery that is to be used. Preferably, when an aerosol route is used the polymeric matrix and composition are encompassed in a surfactant vehicle. The polymeric matrix composition can be selected to have both favorable degradation rates and also to be formed of a material, which is a bioadhesive, to further increase the effectiveness of transfer. The matrix composition also can be selected not to degrade, but rather to release by diffusion over an extended period of time. The delivery system can also be a biocompatible microsphere that is suitable for local, site-specific delivery. Such microspheres are disclosed in Chickering, D. E., et al., Biotechnol. Bioeng., 52: 96-101; Mathiowitz, E., et al., Nature 386: 410-414.

Both non-biodegradable and biodegradable polymeric matrices can be used to deliver the compositions of the invention to the subject. Such polymers may be natural or synthetic polymers. The polymer is selected based on the period of time over which release is desired, generally in the order of a few hours to a year or longer. Typically, release over a period ranging from between a few hours and three to twelve months is most desirable. The polymer optionally is in the form of a hydrogel that can absorb up to about 90% of its weight in water and further, optionally is cross-linked with multivalent ions or other polymers.

Exemplary synthetic polymers which can be used to form the biodegradable delivery system include: polyamides, polycarbonates, poly alky lenes, polyalkylene glycols, poly alky lene oxides, polyalkylene terepthalates, polyvinyl alcohols, polyvinyl ethers, polyvinyl esters, poly-vinyl halides, polyvinylpyrrolidone, polyglycolides, polysiloxanes, polyurethanes and co-polymers thereof, alkyl cellulose, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitro celluloses, polymers of acrylic and methacrylic esters, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxy-propyl methyl cellulose, hydroxybutyl methyl cellulose, cellulose acetate, cellulose propionate, cellulose acetate butyrate, cellulose acetate phthalate, carboxylethyl cellulose, cellulose triacetate, cellulose sulphate sodium salt, poly(methyl methacrylate), poly(ethyl methacrylate),

poly(butylmethacrylate), poly(isobutyl methacrylate), poly(hexylmethacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate), polyethylene, polypropylene, poly(ethylene glycol), poly(ethylene oxide), poly(ethylene terephthalate), poly(vinyl alcohols), polyvinyl acetate, poly vinyl chloride, polystyrene,

polyvinylpyrrolidone, and polymers of lactic acid and glycolic acid, poly anhydrides, poly(ortho)esters, poly(butic acid), poly(valeric acid), and poly(lactide-cocaprolactone), and natural polymers such as alginate and other polysaccharides including dextran and cellulose, collagen, chemical derivatives thereof (substitutions, additions of chemical groups, for example, alkyl, alkylene, hydroxylations, oxidations, and other modifications routinely made by those skilled in the art), albumin and other hydrophilic proteins, zein and other prolamines and hydrophobic proteins, copolymers and mixtures thereof. In general, these materials degrade either by enzymatic hydrolysis or exposure to water in vivo, by surface or bulk erosion.

Methods of Treatment

In one embodiment, the present invention provides a method of enhancing survival or cardiac function in a subject treated with a PI3K inhibitor comprising the step of

administering to the subject an effective amount of OSI-906, preferably as part of a composition additionally comprising a pharmaceutically acceptable carrier. Preferably, this method is employed to treat a subject suffering from or susceptible to a cardiac condition induced by PI3K inhibitor treatment. Other embodiments include any of the methods herein wherein the subject is identified as in need of the indicated treatment.

Another aspect of the invention is the use of OSI-306 in the manufacture of a medicament for enhancing cardiac function in a subject treated with PI3K inhibitor.

Preferably, the medicament is used for treatment or prevention in a subject of a disease, disorder or symptom set forth above.

Kits

The invention provides kits for the treatment or prevention of a cardiac condition associated with PI3K inhibitor treatment. In one embodiment, the kit includes a

pharmaceutical pack comprising an effective amount of OSI-906. Preferably, the compositions are present in unit dosage form. In some embodiments, the kit comprises a sterile container which contains a therapeutic or prophylactic composition; such containers can be boxes, ampules, bottles, vials, tubes, bags, pouches, blister-packs, or other suitable container forms known in the art. Such containers can be made of plastic, glass, laminated paper, metal foil, or other materials suitable for holding medicaments.

If desired compositions of the invention or combinations thereof are provided together with instructions for administering them to a subject having or at risk of developing a cardiac condition associated PI3K inhibitor treatment. The instructions will generally include information about the use of OSI-906 for the treatment or prevention of a cardiac condition associated with PI3K inhibitor treatment. In other embodiments, the instructions include at least one of the following: description of OSI-906; dosage schedule and administration for treatment of a cardiac condition or symptoms thereof; precautions; warnings; indications; counter- indications; overdosage information; adverse reactions; animal pharmacology;

clinical studies; and/or references. The instructions may be printed directly on the container (when present), or as a label applied to the container, or as a separate sheet, pamphlet, card, or folder supplied in or with the container. The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the assay, screening, and therapeutic methods of the invention, and are not intended to limit the scope of what the inventors regard as their invention.

The results presented herein demonstrate that BEZ-induced cardiac hypertrophy is caused by its systematic effects on hepatic gluconeogenesis, hyperglycemia and

hyperinsulinemia. Hyperglycemia is common in PI3K and mTOR inhibitor treated patients. Thus, as described in detail below, blocking of the IGFR/IR in the heart, such as by using OSI-906, prevent BEZ or other PI3K-inhibitor induced cardiac dysfunction.

Example 1: BEZ Induced Cardiac Hypertrophy in Mice

Figure 1 A-Figure IF is a series of bar charts and a photomicrograph of an

immunoblot showing BEZ induced cardiac hypertrophy in mice. Three months old FVB/n female mice (n=10-15 per group) were treated with BEZ (15, 50, 100 mg/kg, daily, five days a week, oral gavage). 5-6 weeks after the treatment the following studies were performed: (Figure 1A) Echocardiography (EDV: end-diastolic volume; ESV: end-systolic volume;

SWth: septum wall thickness; PWth: posterior wall thickness: EF%: ejection fraction; RWth: relative wall thickness); (Figure IB) Hemodynamic measurements (HR: heart rate; LVSP: left ventricular end-systolic pressure; LVW/TL: ratio of LV weight to tibia length; dP/dt: the rate of LV pressure rise or decline); (Figure 1C) Morphology studies for cardiomyocyte cross sectional area and cardiac fibrosis; (Figure ID) Real-time PCR analysis of cardiac hypertrophic marker fetal gene RNA expressions; (Figure 1E-F) Western blot analysis of phosphorylation (activation) of the insulin receptor proteins and downstream signaling pathways.

These results demonstrated that chronic BEZ treatment induced cardiac hypertrophy as demonstrated by a dose-dependent decrease of LV chamber and increase of LV wall thickness, increased LV dP/dtmax and dP/dtmin, increased cardiomyocyte cross sectional area, increased expression of fetal genes. The cardiac hypertrophy induced by BEZ was associated with increased activation of the insulin receptor (IR) and downstream signaling pathways in the heart.

Example 2: Bez Increased Hepatic Gluconeogenesis, Induced Hyperglycemia,

Hyperinsulinemia, and Increased Insulin Receptor Activation in the Heart

Figure 2A-Figure 2B: Fasting glucose: Mice (n=5-8 per point) were treated with BEZ, and fasted. Fasting glucose was measured 8 hours after the treatment. Figure 2C: Glucose tolerance test: Mice (n=5-9 per group) were fasted overnight; GTT was performed 16-18 hours after fasting. Figure 2D: Pyruvate tolerance test: Mice (n=5-8 per group) were fasted overnight; PTT was performed 16-18 hours after fasting. Figure 2E: Real-time PCR of hepatic gluconeogenesis gene expression: Mice (n=5 per group) were treated with BEZ and then fasted and sacrificed at different time points. mRNA expression of

phosphoenolpyruvate carboxykinase (PCK1) and glucose-6-phosphatase (G6PC) in liver were measured by real-time RT-PCR. Figure 2F: Serum insulin: Mice (n=5 per point per group) were treated with BEZ and then fasted and sacrificed at different time points. Serum insulin was measured by ELISA. Figure 2G-Figure 2H: Western blot analysis of IR and downstream signaling pathways: Mice (n=5 per point, per group) were treated with BEZ and then fasted and sacrificed at different time points. Activation of the insulin receptor (IR) and its downstream signaling pathways (PI3K-Akt, mTOR) were assessed by Western blotting using specific antibodies.

These results demonstrated that BEZ increased hepatic gluconeogenesis, which was associated with hyperglycemia, worsened glucose and pyruvate tolerance, hyperinsulinemia and increased activation of the insulin receptor signaling in the heart.

Example 3: Injections of Insulin Worsened, Whereas Blocking IGFR/IR by OSI-906

Alleviated BEZ-Induced Cardiac Hypertrophy

Mice (n=5-10 per group) were treated with BEZ with or without concurrent administration of insulin (0.5, 2, 5, 10, 20 IU, i.p. daily, five days a week) or OSI-906 (0.5 mg/kg, oral gavage, daily, five days a week). Echocardiography and hemodynamic measurements were performed 4-5 weeks after the treatments. Figure 3A: Echocardiography. Figure 3B: Hemodynamic measurements. Figure 3C: Fasting glucose, GTT and PTT.

Figure 3D: Insulin receptor activation in the heart. Figure 3E: Echocardiography in OSI treated mice. Figure 3F: Hemodynamic measurements in OSI treated mice.

These results demonstrated that insulin injections worsened BEZ-induced cardiac dysfunction despite its effects on lowering fasting blood glucose and improving GTT and PTT. These results also demonstrated that OSI-906, an IGFR/IR inhibitor alleviated BEZ- induced cardiac hypertrophy and dysfunction. OTHER EMBODIMENTS

While the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. The patent and scientific literature referred to herein establishes the knowledge that is available to those with skill in the art. All United States patents and published or unpublished United States patent applications cited herein are incorporated by reference. All published foreign patents and patent applications cited herein are hereby incorporated by reference. Genbank and NCBI submissions indicated by accession number cited herein are hereby incorporated by reference. All other published references, documents, manuscripts and scientific literature cited herein are hereby incorporated by reference.

While this invention has been particularly shown and described with references to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.

Claims

What is claimed is:

1. A method of treating phosphoinositide 3-kinase (PI3K) inhibitor-induced

cardiotoxicity comprising:

administering an effective amount of an insulin- like growth factor 1 receptor

(IGFR)/insulin receptor (IR) inhibitor to a subject in need thereof, thereby treating the PI3K inhibitor-induced cardiotoxicity.

2. The method of claim 1, wherein the IGFR/IR inhibitor comprises 3-[8-Amino-l-(2- phenyl-7-quinolyl)imidazo[l,5-a]pyrazin-3-yl]-l-methyl-cyclobutanol (OSI-906).

3. The method of claim 1, wherein cardiac hypertrophy and adverse cardiac remodeling is reduced in the subject. 4. The method of claim 1, wherein the PI3K inhibitor is selected from the group consisting of wortmannin, demethoxyviridin, LY294002, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, GDC-0941, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477, CUDC-907, BYL719, GDC-0032, BGT226, GDC0980, PF4691502, PKI587, and AEZS- 136.

4.

5. The method of claim 4, wherein the PI3K inhibitor comprises 2-methyl-2(4-[3- methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydro-lH-imidazo[4,5-c]quinolin-l- yl]phenyl)propanenitrile (BEZ-235).

6. The method of claim 1, wherein the effective amount of OSI-906 is from 0.5 mg/kg to 1 mg/kg administered daily.

7. The method of claim 1, wherein the subject is identified by echocardiography.

8. The method of claim 1, wherein the OSI-906 is administered prior to, concurrently with, or after administration of the PI3K inhibitor.

9. A method of enhancing cardiac function or increasing survival in a subject treated with a PI3K inhibitor comprising:

administering an effective amount of OSI-906 to the subject, thereby improving cardiac function in the subject.

10. A kit for preventing or treating PI3K inhibitor-induced cardiotoxicity comprising an effective amount of OSI-906 and instructions for use.

11. A pharmaceutical composition comprising an effective amount of BEZ-235 and an effective amount of OSI-906, and instructions for the administration of each.