Methods of treating cardiovascular disease associated with calcification

Abstract

The invention features methods of treating cardiovascular disease associated with calcification (e.g., aortic stenosis, coronary atherosclerosis, PAD, vein graft failure, AV fistula failure, bicuspid aortic valves, myocardial calcification, pericardial calcification, portal vein calcification, calcific uremic arteriopathy, and Hutchinson-Gilford progeria syndrome) by administering to the subject pemafibrate or a pharmaceutically acceptable salt thereof.

Description

[0001]PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 METHODS OF TREATING CARDIOVASCULAR DISEASE ASSOCIATED WITH CALCIFICATION CROSS-REFERENCE TO RELATED APPLICATION This application claims the benefit of U.S. Patent Application Serial No.63 / 611,529, filed on December 18, 2023. The disclosure of the prior application is considered part of (and is incorporated by reference in) the disclosure of this application. BACKGROUND OF THE INVENTION This application relates to methods for treatment of cardiovascular diseases associated with calcification. Cardiovascular calcification, where calcium is deposited on a blood vessel, is a health disorder with increasing prevalence and high morbidity and mortality. Although most calcium is found in teeth and bone, approximately 1% is dissolved in the bloodstream. Cardiovascular calcification occurs when calcium builds up on the walls of the blood vessels, which can restrict blood flow. Early calcification (microcalcification) reduces the mechanical strength of vessels and causes disruption (plaque rupture), leading to acute thrombotic events (e.g., myocardial infarction). Aortic valve calcification causes aortic stenosis (AS) that obstruct the blood flow. Metabolic disorders or organ failures, particularly chronic kidney disease (CKD), accelerate the development of aortic valve calcification. The population of the developed world with AS burden is expected to increase from 2.5 million in 2000 to 4.5 million in 2030. Approximately 10% of individuals diagnosed with AS also present with heart failure (HF) characterized by reduced ejection fraction (HFrEF). General prognosis is poor, as only 50% of patients will survive within 2 years if untreated by aortic valve replacement. Other cardiovascular tissues that develop calcification include vein grafts, a common procedure for atherosclerotic arteries, and arteriovenous fistula (AV fistula), a blood access for hemodialysis. Numerous biological processes are involved in calcification, including matrix remodeling, transcriptional regulation, mitochondrial dysfunction, oxidative stress, calcium and phosphate signaling, endoplasmic reticulum stress, lipid and mineral metabolism, autophagy, inflammation, apoptosis, loss of mineralization inhibition, impaired mineral resorption, cellular senescence and extracellular vesicles that act as precursors of microcalcification. The only available therapeutic options for calcific cardiovascular disease are invasive transcatheter procedures or surgeries that do not fully address the wide spectrum of these conditions; therefore, an urgent need exists for medical options. SUMMARY OF THE INVENTION The invention provides methods for treatment of cardiovascular diseases associated with calcification. In one aspect, the invention generally relates to a method for treating a subject having a cardiovascular disease associated with calcification, including administering to the subject a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof. In some embodiments, patients benefiting from receiving pemafibrate treatment of cardiovascular diseases associated with calcification are identified according to methods known in the art (for example, non-invasive imaging modalities (e.g., ultrasonography (echocardiography), computed tomography (CT), PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 NaF18-positron emission tomography CT (NaF-PET CT),18F-FDG PET (FDG-PET CT), and magnetic resonance imaging (MRI)). In some embodiments, the cardiovascular disease associated with calcification is aortic stenosis (AS). In some embodiments, the treatment results in a decrease in calcification of the aortic valve. In some embodiments, the treatment results in a decrease in inflammation of the aortic valve. In some embodiments, the treatment results in improved open diameter narrowing of the aortic valve. In some embodiments, the treatment results in improved blood flow velocity elevation in the aortic valve. In some embodiments, the treatment results in a decrease in valve leaflet thickness. In some embodiments, the treatment improves cardiac function or prevents heart failure. In some embodiments, the cardiovascular disease associated with calcification is coronary atherosclerosis. In some embodiments, the treatment results in a decrease in calcification of the coronary arteries. In some embodiments, the cardiovascular disease associated with calcification is peripheral artery disease (PAD). In some embodiments, the treatment results in a decrease in calcification of the peripheral arteries. In some embodiments, the cardiovascular disease associated with calcification is vein graft failure. In some embodiments, the treatment results in a decrease in calcification of the vein graft. In some embodiments, the cardiovascular disease associated with calcification is AV fistula failure. In some embodiments, the treatment results in a decrease in calcification of the AV fistula. In some embodiments, the cardiovascular disease associated with calcification is bicuspid aortic valves, myocardial calcification, pericardial calcification, portal vein calcification, calcific uremic arteriopathy, or Hutchinson-Gilford progeria syndrome. In other embodiments, a subject receiving a vein graft is administered a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof. In other embodiments, a subject having an arteriovenous fistula (AV fistula) is administered a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof. In some embodiments, the subject is a human. Advantageously, the inventors found that pemafibrate could be used for the treatment of cardiovascular disease associated with calcification, such as aortic stenosis. The inventors also showed that pemafibrate can reduce pro-inflammatory calcification of valvular interstitial cells (VICs) and that plasma of patients treated with pemafibrate reduces valvular cell calcification in vitro. Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims. Definitions As used herein, “disease” is used in this disclosure to mean, and is used interchangeably with, the terms disease, condition, or illness, unless otherwise indicated. As used herein, “therapeutically effective amount” refers to an amount of pemafibrate or a pharmaceutically acceptable salt thereof sufficient to treat a cardiovascular disease associated with calcification in a subject. In some embodiments, the therapeutically effective amount of pemafibrate or a PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 pharmaceutically acceptable salt thereof may be an amount sufficient to treat aortic stenosis. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may decrease calcification of the aortic valve. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may decrease inflammation of the aortic valve. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may improve open diameter narrowing of the aortic valve. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may improve blood flow velocity elevation in the aortic valve. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may decrease aortic valve leaflet thickness. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may improve cardiac function or prevent heart failure. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may be an amount sufficient to treat coronary atherosclerosis. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may decrease calcification of the coronary arteries. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may be an amount sufficient to treat PAD. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may decrease calcification of the peripheral arteries. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may be an amount sufficient to treat vein graft failure. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may decrease calcification of the vein graft. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may be an amount sufficient to treat AV fistula failure. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may decrease calcification of the AV fistula. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may be an amount sufficient to treat bicuspid aortic valves, myocardial calcification, pericardial calcification, portal vein calcification, calcific uremic arteriopathy, or Hutchinson-Gilford progeria syndrome. In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may be an amount sufficient to treat a vein graft (e.g., a graft demonstrating signs of calcification). In some embodiments, the therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof may be an amount sufficient to treat an AV fistula (e.g., an AV fistula demonstrating signs of calcification). As used herein, “pharmaceutically acceptable” means that which is useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable and includes that which is acceptable for veterinary use as well as human pharmaceutical use. “Pharmaceutically acceptable salts” means salts that are pharmaceutically acceptable, as defined above, and which possess the desired pharmacological activity. As used herein, the term “treating” refers to partially or completely alleviating, ameliorating, improving, relieving, delaying onset of, inhibiting progression of, reducing severity of, and / or reducing PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 incidence of one or more symptoms or features of a cardiovascular disease associated with calcification. Treatment may be administered to a subject who does not exhibit signs of the cardiovascular disease and / or to a subject who exhibits only early signs of the cardiovascular disease for the purpose of decreasing the risk of developing pathology associated with the cardiovascular disease. As used herein, the terms “subject” and “patient” are interchangeable and refer to an organism that receives treatment for a particular disease or condition as described herein. In general, the subject or patient is human. As used herein, “calcification” refers to the deposition of calcium on the wall of a blood vessel. Cardiovascular calcification occurs when calcium builds up on the walls of the blood vessels, which can restrict blood flow. If not treated, calcification can cause a blood clot that blocks blood flow. Cardiovascular calcification is an active process involving numerous biological processes, including matrix remodeling, transcriptional regulation, mitochondrial dysfunction, oxidative stress, calcium and phosphate signaling, endoplasmic reticulum stress, lipid and mineral metabolism, autophagy, inflammation, apoptosis, loss of mineralization inhibition, impaired mineral resorption, cellular senescence and extracellular vesicles that act as precursors of microcalcification. In some embodiments, calcification includes microcalcification, which is identified according to standard methods such as those disclosed herein. As used herein, the term “cardiovascular disease associated with calcification” refers to any cardiovascular disease that has been linked to calcification of a blood vessel. The cardiovascular disease associated with calcification may include, but is not limited to, aortic stenosis, coronary atherosclerosis, PAD, vein graft failure, AV fistula failure, bicuspid aortic valves, myocardial calcification, pericardial calcification, portal vein calcification, calcific uremic arteriopathy, and Hutchinson-Gilford progeria syndrome. As used herein, “aortic stenosis” (AS) refers to a condition where the aortic valve narrows and blood cannot flow normally. The cause of AS is deposition of calcium in the aortic valve. As blood repeatedly flows over the aortic valve, calcium deposits build up on the heart valves, leading to narrowing and thickening of the aortic valve. Once valve leaflet mobility is reduced by calcification, the heart’s left ventricle pumps harder to push blood through the narrowed aortic valve. The pressure in the left ventricle increases and a pressure difference can be measured between the left ventricle and the aorta. To compensate for the increasing resistance at the aortic valve, the muscles of the left ventricle thicken to maintain pump function and cardiac output. This muscle thickening causes a stiffer heart muscle which requires higher pressures in the left atrium and the blood vessels of the lungs to fill the left ventricle. As the disease progresses, the increasing pressure eventually causes the left ventricle to dilate, leading to a decrease in cardiac function and heart failure. Approximately 10% of individuals diagnosed with AS also present with heart failure (HF) characterized by reduced ejection fraction (HFrEF). Symptomatic AS, if untreated by aortic valve replacement (AVR), drastically increases the risk of mortality to 50% within 2 years due to HF. As used herein, the term “decrease in calcification of the aortic valve” refers to a decrease in the amount of calcification of the aortic valve after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Decreased PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 calcification of the aortic valve can be measured by noninvasive imaging modalities, including but not limited to, ultrasonography (echocardiography), CT, NaF-PET CT, FDG-PET CT and MRI. As used herein, the term “decrease in inflammation of the aortic valve” refers to a decrease in the amount of inflammation of the aortic valve after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Decreased inflammation of the aortic valve can be measured using fluorodeoxyglucose (FDG)-PET. As used herein, “open diameter narrowing” refers to the narrowing of the aortic valve that occurs in aortic stenosis due to calcification and thickening of the leaflets. Open diameter narrowing can be measured using ultrasonography (echocardiography). As used herein, the term “improved open diameter narrowing of the aortic valve” refers to an improvement in the open diameter narrowing after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Open diameter narrowing is improved if the diameter of the aortic valve increases. Open diameter narrowing can be measured using ultrasonography (echocardiography). As used herein, “blood flow velocity elevation” refers to the increased blood flow velocity in the aortic valve that occurs in aortic stenosis. As used herein, the term “improved blood flow velocity elevation in the aortic valve” refers to an improvement in blood flow velocity elevation after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Blood flow velocity elevation is improved if the blood flow velocity decreases. Blood flow velocity elevation can be measured using ultrasonography (echocardiography). As used herein, “valve leaflet thickness” refers to the thickness of the aortic valve. In aortic stenosis, the thickness of the aortic valve increases due to inflammation and calcification. As used herein, the term “decrease in aortic valve leaflet thickness” refers to a decrease in the aortic valve leaflet thickness after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Aortic valve leaflet thickness can be measured using ultrasonography (echocardiography) or CT. As used herein, “coronary atherosclerosis” refers to a condition where plaque (fatty deposits) build up in the coronary arteries. These deposits are made up of cholesterol, fatty substances, cellular waste products, calcium and fibrin (a clotting material in the blood). As plaque builds up, the walls of the coronary arteries thicken and stiffen. This narrows the channel within the coronary arteries, reducing blood flow and lessening the amount of oxygen and other nutrients reaching the body. As used herein, “decrease in calcification of the coronary arteries” refers to a decrease in calcification of the coronary arteries after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Calcification of the coronary arteries may be measured using coronary calcium scoring by CT. As used herein, “peripheral artery disease” (PAD) refers to a condition where the peripheral arteries that carry blood away from the heart to other parts of the body become narrower. The most common type is lower-extremity PAD, in which blood flow is reduced to the legs and feet. Upper-extremity PAD (arms, hands and fingers) is less common but affects about 10% of the population. PAD is caused PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 by atherosclerosis, the buildup of fatty plaque, including calcium, in the arteries that narrows and blocks them. Among the peripheral arteries most commonly blocked due to calcium buildup include iliac arteries in the pelvis, superficial femoral artery or SFA in the thigh, and infrapopliteal artery below the knee. As used herein, “decrease in calcification of the peripheral arteries” refers to a decrease in calcification of the peripheral arteries after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Calcification of the peripheral arteries can be measured using ultrasonography or CT. As used herein, “vein graft” refers to the conduit used in coronary artery bypass graft surgery where the great saphenous vein (the large vein running up the length of the leg) is grafted onto arteries that feed the heart to bypass clogged vessels and restore blood flow. After grafting, implanted veins remodel to become more arterial. As used herein, “vein graft failure” refers to a condition where the inner walls of the vein become too thick following coronary artery bypass graft surgery, slowing down or blocking the blood flow that the graft was intended to restore. Calcification in the wall of the vein graft is associated with an increased incidence of vein graft lesions and failure. As used herein, “decrease in calcification of the vein graft” refers to a decrease in calcification of the vein graft after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Calcification of the vein graft can be measured using ultrasonography. As used herein, “arteriovenous fistula” (AV fistula) refers to a surgical connection that is made between an artery and a vein for dialysis access. The access point must be durable enough to withstand dialysis treatment several times a week without collapsing. An AV fistula is typically located in the arm, however, if necessary, it can be placed in the leg. The increased flow and pressure cause the veins to enlarge. The enlarged veins will be capable of delivering the amount of blood flow necessary to provide an adequate hemodialysis treatment. AV fistulas are the preferred vascular access for long-term dialysis because they last longer than any other dialysis access types, are less prone to infection and clotting, and can be relied upon for predictable performance. As used herein, “AV fistula failure” refers to a condition where, following surgical AV fistula creation, the AV fistula is never usable or fails within the first three months of its use. Calcification of the AV fistula is associated with AV fistula failure. As used herein, “decrease in calcification of the AV fistula” refers to a decrease in calcification of the AV fistula after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Calcification of the AV fistula can be measured using ultrasonography. Patency of the AV fistula can also be assessed by physical examinations. As used herein, “heart failure” means an abnormality of cardiac function where the heart does not pump blood at the rate needed for the requirements of metabolizing tissues. Heart failure includes a wide range of disease states such as congestive heart failure, myocardial infarction, tachyarrhythmia, familial hypertrophic cardiomyopathy, ischemic heart disease, idiopathic dilated cardiomyopathy, myocarditis and the like. Heart failure can be caused by any number of factors, including, without limitation, ischemic, congenital, rheumatic, or idiopathic forms. PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 As used herein, “improves cardiac function” refers to an improvement in cardiac function after treatment with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to before treatment. Cardiac function may be measured using ultrasonography (echocardiography). As used herein, “cardiac function” refers to the ability of the heart to meet the metabolic demands of the body. Cardiac function may be determined through exercise capacity, cardiac ejection volume, left ventricular end diastolic pressure, pulmonary capillary wedge pressure, cardiac output, cardiac index, pulmonary artery pressures, left ventricular end systolic and diastolic dimensions, left and right ventricular wall stress, and wall tension. Assessment tests for cardiac functions are well known to a skilled practitioner. Exemplary tests for cardiac function include, but not limited to, echocardiography, electrocardiogram, X-ray, magnetic resonance imaging, coronary catheterization, and heart CT scan. As used herein, “prevents heart failure” refers to the prevention of heart failure with a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof compared to without treatment. Heart failure can be measured using ultrasonography (echocardiography). Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. For any term present in the art which is identical to any term expressly defined in this disclosure, the term's definition presented in this disclosure will control in all respects. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods and materials are described herein. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the study design of an aortic stenosis (AS) mouse model with and without pemafibrate treatment. Figure 2 shows body weight and food intake in mice treated with pemafibrate on a normal or high fat diet in an AS mouse model. Figure 3 shows that pemafibrate decreased triglycerides but did not affect total cholesterol in mice with AS fed a normal diet (ND) or a high fat diet (HFD). Figure 4 shows that pemafibrate improved open diameter narrowing in AS mouse model. Figure 5 shows that pemafibrate improved blood flow velocity elevation in the aortic valve of a mouse model of AS. Figure 6 shows that pemafibrate did not change relative heart weight in a mouse model of AS. Figure 7 shows that pemafibrate reduced calcification of aortic valves in a mouse model of AS. Figure 8 shows that pemafibrate reduced inflammation of aortic valves in a mouse model of AS. Figure 9 shows that pemafibrate decreased valve leaflet thickness and macrophage infiltration in aortic valves. Figures 10A-10B show that pemafibrate decreased IL-12 and S100A9 protein expression in aortic valves. Figure 11 shows that pemafibrate suppressed IL-6, IL-12, S100A8, and S100A9 gene expression in THP-1-derived macrophage-like cells. PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 Figure 12 shows that pemafibrate-treated THP-1 macrophage-derived conditioned media promotes anti-calcification effects in hVICs. DETAILED DESCRIPTION OF THE INVENTION The invention provides methods for treatment of cardiovascular diseases associated with calcification, involving administering to the subject a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof. Pemafibrate, whose chemical name is (2R)-2-[3-({1,3- benzoxazol-2-yl[3-(4-methoxyphenoxy)propyl]amino}methyl)phenoxy]butanoic acid, is a highly selective Peroxisome proliferator-activated receptor (PPAR)-alpha modulator. PPARα is expressed abundantly in skeletal muscle, liver, intestine, kidney, and heart, and regulates fatty acid (FA) transport, esterification, and oxidation. Activation of PPARα is known to limit the inflammatory response. The studies presented herein demonstrate that pemafibrate or a pharmaceutically acceptable salt thereof may be used for the treatment of cardiovascular diseases associated with calcification. The invention is based, at least in part, on the discovery that pemafibrate suppresses calcification deposition. In some embodiments, pemafibrate inhibits the process of microcalcification deposition, which is the initial stage of calcification accumulation. In the Examples below, we describe the efficacy of pemafibrate or a pharmaceutically acceptable salt thereof in treating cardiovascular diseases associated with calcification. Treatment with pemafibrate or a pharmaceutically acceptable salt thereof decreased calcification, inflammation, and thickness of the aortic valve, and improved both open diameter narrowing and blood flow velocity elevation. Overall, the disclosure demonstrates that treatment with pemafibrate or a pharmaceutically acceptable salt thereof improves cardiac function and prevents heart failure. I. Pemafibrate Pemafibrate, whose chemical name is (2R)-2-[3-({1,3-benzoxazol-2-yl[3-(4- methoxyphenoxy)propyl]amino}methyl)phenoxy]butanoic acid, is a highly selective PPAR-alpha modulator. The structure of pemafibrate is given below. PPARα is expressed abundantly in skeletal muscle, liver, intestine, kidney, and heart, and regulates fatty acid (FA) transport, esterification, and oxidation. Activation of PPARα is known to limit the inflammatory response. PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 In some embodiments, any pharmaceutically acceptable salt of pemafibrate may be administered. The pharmaceutically acceptable salt may include acid addition salts and base addition salts. Specific examples of the acid addition salts include acid addition salts with inorganic acids, such as hydrochlorides, hydrobromides, hydroiodides, sulfate salts, nitrate salts and phosphate salts; and acid addition salts with organic acids, such as benzoate salts, methanesulfonate salts, ethanesulfonate salts, benzenesulfonate salts, p-toluenesulfonate salts, maleate salts, fumarate salts, tartrate salts, citrate salts and acetate salts. Specific examples of the base addition salts include metal salts such as sodium salts, potassium salts, lithium salts, calcium salts and magnesium salts; salts with amines such as ammonia, trimethylamine, triethylamine, pyridine, collidine and lutidine; and base addition salts with organic bases such as lysine, arginine, cinchonine and cinchonidine. In some embodiments, the pharmaceutically acceptable salt may be any pemafibrate salt described in US20220372006A1. II. Pharmaceutical Compositions Acceptable carriers and excipients in the pharmaceutical compositions are nontoxic to recipients at the dosages and concentrations employed. Acceptable carriers and excipients may include buffers such as phosphate, citrate, HEPES, and TAE, antioxidants such as ascorbic acid and methionine, preservatives such as hexamethonium chloride, octadecyldimethylbenzyl ammonium chloride, resorcinol, and benzalkonium chloride, proteins such as human serum albumin, gelatin, dextran, and immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acids such as glycine, glutamine, histidine, arginine, and lysine, and carbohydrates such as glucose, mannose, sucrose, and sorbitol. Pharmaceutical compositions of the invention can be administered parenterally in the form of an injectable formulation. Pharmaceutical compositions for injection can be formulated using a sterile solution or any pharmaceutically acceptable liquid as a vehicle. Pharmaceutically acceptable vehicles include, but are not limited to, sterile water, physiological saline, and cell culture media (e.g., Dulbecco’s Modified Eagle Medium (DMEM), α-Modified Eagles Medium (α-MEM), F-12 medium). Formulation methods are known in the art, see e.g., Banga (ed.) Therapeutic Peptides and Proteins: Formulation, Processing and Delivery Systems (3rd ed.) Taylor & Francis Group, CRC Press (2015). The pharmaceutical compositions of the invention may be prepared in microcapsules, such as hydroxylmethylcellulose or gelatin-microcapsule and poly-(methylmethacrylate) microcapsule. The pharmaceutical compositions of the invention may also be prepared in other drug delivery systems such as liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules. Such techniques are described in Remington: The Science and Practice of Pharmacy 22ndedition (2012). The pharmaceutical compositions to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes. The pharmaceutical compositions of the invention may also be prepared as a sustained-release formulation. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the polypeptides of the invention. Examples of sustained release matrices include polyesters, hydrogels, polylactides, copolymers of L-glutamic acid and γ ethyl-L- glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON DEPOTTM, and poly-D-(-)-3-hydroxybutyric acid. Some sustained-release formulations PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 enable release of molecules over a few months, e.g., one to six months, while other formulations release pharmaceutical compositions of the invention for shorter time periods, e.g., days to weeks. The pharmaceutical composition may be formed in a unit dose form as needed. The amount of active component, e.g., pemafibrate or a pharmaceutically acceptable salt thereof, included in the pharmaceutical preparations is such that a suitable dose within the designated range is provided (e.g., a dose within the range of 0.01-100 mg / kg of body weight). III. Dosing and Administration Pemafibrate, or a pharmaceutically acceptable salt thereof, can be administered to a subject (e.g., a human) by a variety of routes, such as orally, transdermally, subcutaneously, intranasally, intravenously, intramuscularly, intraocularly, intratumorally, parenterally, topically, intrathecally and intracerebroventricularly, for the treatment of a cardiovascular disease associated with calcification. Pemafibrate, or a pharmaceutically acceptable salt thereof, may be administered, e.g., by injection, such as by intravenous, intramuscular, intraperitoneal, or subcutaneous injection. Typically, pemafibrate is administered in tablet form. A daily dose of a therapeutic composition of pemafibrate or a pharmaceutically acceptable salt thereof may be administered as a single dose or as two, three, four, five, six or more doses administered separately at appropriate intervals throughout the day, week, month, or year, or as needed, optionally, in unit dosage forms. While it is possible for pemafibrate, or a pharmaceutically acceptable salt thereof, to be administered alone, it may also be administered as a pharmaceutical formulation in combination with excipients, carriers, and optionally, additional therapeutic agents. The effective dose of pemafibrate, or a pharmaceutically acceptable salt thereof, can range, for instance, from about 0.0001 to about 100 mg / kg of body weight per single (e.g., bolus) administration, multiple administrations or continuous administration (e.g., a continuous infusion), or to achieve a serum concentration of 0.0001-5000 μg / mL serum concentration per single (e.g., bolus) administration, multiple administrations or continuous administration (e.g., continuous infusion), or any effective range or value therein depending on the condition being treated, the route of administration and the age, weight, and condition of the subject. In certain embodiments, each dose can range from about 0.0001 mg to about 500 mg / kg of body weight. For instance, a pharmaceutical composition described herein may be administered in a daily dose in the range of 0.0001-100 mg / kg (body weight). The dose may be administered one or more times (e.g., 2-10 times) per day, week, month, or year to a subject (e.g., a human) in need thereof. In some embodiments, pemafibrate may be administered at a dose of about 0.1 mg / day to about 0.4 mg / day. In some embodiments, pemafibrate may be administered at a dose of up to 0.2 mg tablets, twice daily. IV. Methods of Treatment The invention is based on the discovery that pemafibrate, or a pharmaceutically acceptable salt thereof, can be used to treat cardiovascular diseases associated with calcification (e.g., aortic stenosis, coronary atherosclerosis, PAD, vein graft failure, AV fistula failure, bicuspid aortic valves, myocardial PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 calcification, pericardial calcification, portal vein calcification, calcific uremic arteriopathy, and Hutchinson- Gilford progeria syndrome). In some embodiments, subjects have been identified as having a cardiovascular disease associated with calcification using standard methods known in the art, such as by non-invasive imaging modalities (e.g., ultrasonography (echocardiography), computed tomography (CT), NaF18-positron emission tomography CT (NaF-PET CT), and magnetic resonance imaging). Ultrasonography (echocardiography), computed tomography (CT), and NaF18-positron emission tomography CT (NaF-PET CT) are the most commonly used methods for noninvasively identifying patients with aortic valve calcification, aortic stenosis (AS), aortic valve inflammation, and vascular calcification. A reduction of calcification may be visualized by these imaging modalities. Calcification usually develops slowly and clinical trials with primary endpoints would take a few years; however, for example, in patients with the end-stage renal disease (ESRD), particularly on hemodialysis, calcification in the arteries or aortic valves would develop withing several months. Therefore, monitoring disease treatment using imaging could result in a shorter trial. Once identified, such subjects are treated with pemafibrate as disclosed herein. In some embodiments, pemafibrate can be used to inhibit or reduce the process of microcalcification, which is the initial stage of calcification accumulation. Emerging imaging modalities may be used to predict the presence of microcalcification in patients (Hutcheson et al. Curr Opin Lipidol.2014 Oct;25(5):327-32). In addition, 18F-sodium fluoride positron emission tomography may quantify medial microcalcification and could be a feasible noninvasive imaging modality for identifying microcalcification in patients (Fletcher et al., Arterioscler Thromb Vasc Biol.2022 Aug;42(8):1048-1059.). In some embodiments, the subject is a human. Aortic Stenosis In some embodiments, pemafibrate, or a pharmaceutically acceptable salt thereof, can be used to treat aortic stenosis. Aortic stenosis is a condition where the aortic valve narrows and blood cannot flow normally. The cause of aortic stenosis is deposition of calcium in the aortic valve, leading to narrowing of the aortic valve. Metabolic disorders or organ failures, particularly CKD, accelerate the development of aortic valve calcification. Once valve leaflet mobility is reduced by calcification, the heart’s left ventricle pumps harder to push blood through the narrowed aortic valve. The pressure in the left ventricle increases and a pressure difference can be measured between the left ventricle and the aorta. To compensate for the increasing resistance at the aortic valve, the muscles of the left ventricle thicken to maintain pump function and cardiac output. This muscle thickening causes a stiffer heart muscle which requires higher pressures in the left atrium and the blood vessels of the lungs to fill the left ventricle. As the disease progresses, the increasing pressure eventually causes the left ventricle to dilate, leading to a decrease in cardiac function and heart failure. Without treatment, the average life expectancy after the onset of heart failure due to aortic stenosis is between 18 to 24 months. No drug therapy exists to treat aortic stenosis as randomized controlled trials targeting low- density lipoprotein (LDL) cholesterol or mediators of bone metabolism have failed. Due to insufficient mechanistic insight, aortic stenosis is currently treated only at advanced stages by costly surgical AV PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 replacement (SAVR) or transcatheter AV implantation (TAVI), while medical therapies remain unavailable. As such, there is an urgent need for drug therapies for aortic stenosis. In some embodiments, the treatment results in a decrease in calcification of the aortic valve. Deposition of calcium on the aortic valve, where serum calcium attaches to the valve surface and binds to the leaflet to form nodules, is considered to be the root cause of aortic stenosis. In some embodiments, the treatment results in improved open diameter narrowing of the aortic valve. In aortic stenosis, the aortic valve narrows due to thickening and calcification of the leaflets. In this embodiment, treatment with pemafibrate or a pharmaceutically acceptable salt thereof reduces the narrowing of the aortic valve. In some embodiments, the treatment results in improved blood flow velocity elevation in the aortic valve. In aortic stenosis, the blood flow velocity in the aortic valve is increased. In this embodiment, treatment with pemafibrate or a pharmaceutically acceptable salt thereof lowers blood flow velocity. In some embodiments, the treatment results in a decrease in aortic valve leaflet thickness. In aortic stenosis, the thickness of the aortic valve increases due to inflammation and calcification. In some embodiments, the treatment results in a decrease in inflammation in the aortic valve. Inflammation of the aortic valve contributes to its thickening and promotes calcification. In some embodiments, the treatment prevents heart failure. Heart failure is an abnormality of cardiac function where the heart does not pump blood at the rate needed for the requirements of metabolizing tissues. Heart failure includes a wide range of disease states such as congestive heart failure, myocardial infarction, tachyarrhythmia, familial hypertrophic cardiomyopathy, ischemic heart disease, idiopathic dilated cardiomyopathy, myocarditis and the like. The heart failure can be caused by any number of factors, including, without limitation, ischemic, congenital, rheumatic, or idiopathic forms. In some embodiments, the treatment improves cardiac function. Cardiac function, the ability of the heart to meet the metabolic demands of the body, may be determined through exercise capacity, cardiac ejection volume, left ventricular end diastolic pressure, pulmonary capillary wedge pressure, cardiac output, cardiac index, pulmonary artery pressures, left ventricular end systolic and diastolic dimensions, left and right ventricular wall stress, and wall tension. Assessment tests for cardiac functions are well known to a skilled practitioner. Exemplary tests for cardiac function include, but not limited to, echocardiography, electrocardiogram, X-ray, magnetic resonance imaging, coronary catheterization, and heart CT scan. Coronary atherosclerosis In some embodiments, pemafibrate, or a pharmaceutically acceptable salt thereof, can be used to treat coronary atherosclerosis. Coronary atherosclerosis is a condition where plaque (fatty deposits) build up in the coronary arteries. These deposits are made up of cholesterol, fatty substances, cellular waste products, calcium and fibrin (a clotting material in the blood). As plaque builds up, the walls of the coronary arteries thicken and stiffen. This narrows the channel within the coronary arteries, reducing blood flow and lessening the amount of oxygen and other nutrients reaching the body. In addition, early calcification (microcalcification) reduces mechanical stability or strength of atherosclerotic lesions and PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 causes physical disruption (plaque rupture), leading to acute thrombotic complications including myocardial infarction. In some embodiments, the treatment results in a decrease in calcification or microcalcification of the coronary arteries. Deposition of calcium on the coronary arteries, where serum calcium attaches to the artery surface, is associated with coronary atherosclerosis. Peripheral Artery Disease (PAD) In some embodiments, pemafibrate, or a pharmaceutically acceptable salt thereof, can be used to treat PAD. PAD is a condition where the peripheral arteries that carry blood away from the heart to other parts of the body become narrower. The most common type is lower-extremity PAD, in which blood flow is reduced to the legs and feet. Upper-extremity PAD (arms, hands and fingers) is less common but affects about 10% of the population. PAD is caused by atherosclerosis, the buildup of fatty plaque, including calcium, in the arteries that narrows and blocks them. Among the arteries most commonly blocked due to calcium buildup include iliac arteries in the pelvis, superficial femoral artery or SFA in the thigh, and infrapopliteal artery below the knee. In some embodiments, the treatment results in a decrease in calcification of the peripheral arteries (e.g., iliac arteries in the pelvis, superficial femoral artery or SFA in the thigh, and infrapopliteal artery below the knee). Deposition of calcium on the peripheral arteries, where serum calcium attaches to the artery surface, is associated with PAD. In some embodiments, the treatment results in a decrease in inflammation in PAD. Inflammation contributes to the development of PAD. Vein graft failure A vein graft is the conduit used in coronary artery bypass graft surgery where the great saphenous vein (the large vein running up the length of the leg) is grafted onto arteries that feed the heart to bypass clogged vessels and restore blood flow. After grafting, implanted veins remodel to become more arterial. In some embodiments, pemafibrate, or a pharmaceutically acceptable salt thereof, can be used to treat vein graft failure. Vein graft failure is a condition where the inner walls of the vein become too thick following coronary artery bypass graft surgery, slowing down or sometimes even blocking the blood flow that the graft was intended to restore. Calcification in the wall of the vein graft is associated with an increased incidence of vein graft lesions and failure. In some embodiments, the treatment results in a decrease in calcification of the vein graft. Deposition of calcium on the peripheral arteries, where serum calcium attaches to the vein surface, is associated with vein graft failure. In some embodiments, the treatment results in a decrease in inflammation of the vein graft. Inflammation contributes to the development of vein graft disease and failure. PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 Arteriovenous Fistula (AV) Failure An AV fistula is a surgical connection that is made between an artery and a vein for dialysis access. The access point must be durable enough to withstand dialysis treatment several times a week without collapsing. An AV fistula is typically located in the arm, however, if necessary it can be placed in the leg. The increased flow and pressure cause the veins to enlarge. The enlarged veins will be capable of delivering the amount of blood flow necessary to provide an adequate hemodialysis treatment. AV fistulas are the preferred vascular access for long-term dialysis because they last longer than any other dialysis access types, are less prone to infection and clotting, and can be relied upon for predictable performance. In some embodiments, pemafibrate, or a pharmaceutically acceptable salt thereof, can be used to treat AV fistula failure. AV fistula failure is a condition where, following surgical AV fistula creation, the AV fistula is never usable or fails within the first three months of its use. Calcification of the AV fistula is associated with AV fistula failure. In some embodiments, the treatment results in a decrease in calcification of the AV fistula. Deposition of calcium on the AV fistula, where serum calcium attaches to the AV fistula surface, is associated with AV fistula failure. In some embodiments, the treatment results in a decrease in inflammation of the AV fistula. Inflammation contributes to the development of AV fistula failure. EXAMPLES The following examples are put forth so as to provide those of ordinary skill in the art with a description of how the compositions and methods claimed herein are performed, made, and evaluated, and are intended to be purely exemplary described herein and are not intended to limit the scope of what the inventors regards as their invention. Example 1. Study design A mouse model of aortic stenosis (AS mice) was established based on the method published by Honda et al, Arterioscler Thromb Vasc Biol 2014, 34(2): 270-8. Ldlr- / - mice were anesthetized by inhalation of 1% isoflurane. The right carotid artery was exposed by dissection and then ligated. A metal guide wire (ASHI INTECC MIRACLEBros 6) was introduced into the left ventricle under echocardiographic guidance. To damage the aortic valve, the wire’s tip was positioned just below the aortic valve level and manipulated with 20 times strokes and 30 times rotation. The wire then was removed, and the carotid artery was ligated. The animals were randomly assigned into four groups (N = 10-12 per group; 15 weeks). Group 1: normal diet, control: Group 2: normal diet with 1.8 ppm pemafibrate; Group 3: High Fat Diet (HFD), control; Group 4: HFD with 1.8 ppm pemafibrate. Animals were monitored using echocardiography for cardiac function before surgery, and 8 or 15 weeks after surgery. Aortic valve open diameter and peak velocity were measured in the parasternal long axis view with a pulse wave doppler. At 15 weeks after surgery, to visualize calcification and inflammation, OsteoSense680 (calcification tracer) and MMPSense750 (inflammation tracer) molecular near-infrared fluorescent (NIRF) PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 imaging agents were injected 24 hours before sacrifice. Aortic valve tissue was collected after saline perfusion and subjected to imaging analysis to visualize calcification and inflammation / MMP activity. Aortic valves were embedded in OCT compound and cryo-sectioned into 7 µm slices. Sections were stained with hematoxylin and eosin for overall morphology according to routine protocols. Macrophage infiltration in stenotic aortic valves was assessed using CD68 antibody. The pro-inflammatory protein expression was evaluated using IL-12 and S100A9 antibodies. Hematoxylin and eosin staining and immunohistochemistry images were taken using NIS elements software (Nikon) and analyzed using ImageJ software. Human valvular interstitial cells (hVICs) were isolated from calcified aortic valve leaflets obtained from patients undergoing aortic valve replacement at the Brigham and Women’s Hospital (Boston, MA, USA; IRB protocol # 2011P001703) refer to the method of Clift et al. To determine the amount of calcification, the cells were cultured with normal media (NM) (5% FBS and 1% P / S), osteogenic media (OM) (5% FBS, 1% P / S, 10 nM dexamethasone, ascorbic acid (50 μg / ml), and 10 mM β- glycerolphosphate) or THP-1 macrophage-like cells-derived conditioned media, which was refreshed twice per week. Afterward, the amount of calcification in hVICs was evaluated by Alizarin Red staining. THP-1 monocytes were differentiated into macrophage-like cells using phorbol 12-myristate 13- acetate (PMA, 100ng / mL for 48 hours followed by 24 hours recovery culture without PMA). The macrophage-like cells-derived conditioned media was harvested 48 hours after the replacement of the media containing with or without pemafibrate. The conditioned media (DMEM / conditioned media mixed with an equal amount of each) was treated to hVICs. Gene expression analysis was performed 48 hours after pemafibrate treatment in THP-1-derived macrophage-like cells. Example 2. Body weight and food intake of AS mice treated with pemafibrate or the control Pemafibrate suppressed body weight gain in the groups fed a high fat diet. The animals receiving valve injury surgery to induce aortic stenosis showed no abnormal bodyweight and similar food intake (Figure 2). Example 3. Cholesterol and triglyceride levels of AS mice treated with pemafibrate or the control Pemafibrate decreased triglycerides but did not affect total cholesterol in mice with aortic stenosis (Figure 3). Example 4. Open diameter narrowing of AS mice treated with pemafibrate or the control Pemafibrate improved narrowed open diameter of aortic valve at 15 weeks after surgery in both normal and HFD conditions (Figure 4).15 weeks post-surgery, mice receiving the control had a significantly lower (p<0.0001) aortic valve open diameter compared to pre-surgery, indicating the presence of aortic stenosis. Mice treated with pemafibrate had a significantly larger (p<0.01) aortic valve open diameter than mice receiving the control, suggesting that pemafibrate treatment slowed the development of aortic stenosis. PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 Example 5. Blood flow velocity of AS mice treated with pemafibrate or the control At 8 and 15 weeks after surgery in both normal and HFD conditions, mice receiving the control had a significantly higher (p<0.001, normal diet; p<0.0001; HFD diet) aortic valve peak flow velocity compared to pre-surgery, indicating the presence of aortic stenosis. Mice receiving pemafibrate had significantly lower blood flow velocity compared to mice receiving the control at 8 and 15 weeks after surgery in both normal and HFD conditions (Figure 5). These results demonstrate that, while HFD diet accelerated aortic stenosis, pemafibrate decreased development of aortic stenosis. Example 6. Heart weight of AS mice treated with pemafibrate or the control While mice treated with pemafibrate had lower body weight than mice receiving the control 15 weeks post-surgery in both normal and HFD conditions, there was no difference in relative heart weight (Figure 6). Example 7. Calcification of aortic valves of AS mice treated with Pemafibrate or the control Calcification of aortic valves at 15 weeks post-surgery was visualized using near infra-red fluorescent (NIRF) imaging probes. In HFD conditions, pemafibrate treatment significantly decreased (p<0.01) calcification signal in the aortic valve compared to the control (Figure 7), suggesting that pemafibrate suppresses calcification processes in aortic valves. These results demonstrate that, while HFD diet accelerated aortic stenosis and aortic valve calcification, pemafibrate decreased development of aortic stenosis and aortic valve calcification. Example 8. Inflammation of aortic valves of AS mice treated with pemafibrate or the control Matrix metalloproteinase (MMP) activity at 15 weeks post-surgery in aortic valves was visualized using near infra-red fluorescent imaging probes. Pemafibrate treatment significantly decreased MMP activity in both normal and HFD conditions compared to treatment with the control (Figure 8), suggesting that pemafibrate suppresses inflammatory processes in the aortic valve. Example 9. Valve leaflet thickness and macrophage infiltration of AS mice treated with pemafibrate or the control Immunohistochemistry using anti-CD68 antibody detecting macrophages showed that pemafibrate decreased macrophage infiltration and leaflet thickness of the aortic valves compared to treatment with the control (Figure 9). Example 10. Pemafibrate decreased IL-12 and S100A9 protein expression in aortic valves. The results of immunohistochemistry using anti-IL-12 and anti-S100A9 detecting pro- inflammatory proteins indicated that pemafibrate suppressed expression of these pro-inflammatory factors in the aortic valve (Figures 10A-10B). PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 Example 11. Pemafibrate suppressed IL-6, IL-12, S100A8, and S100A9 gene expression in THP-1- derived macrophage-like cells To examine anti-inflammatory effects by pemafibrate, THP-1-derived macrophage-like cells were treated with pemafibrate (Figure 11). Pemafibrate inhibited pro-inflammatory gene expression in a dose- dependent manner, which was consistent with the immunohistochemistry results. Example 12. Pemafibrate-treated THP-1 macrophage-derived conditioned media showed anti- calcification effects in hVICs It was hypothesized that pemafibrate exerted the anti-calcification effects via proinflammatory mechanisms since pemafibrate significantly inhibited the pro-inflammatory gene expression in THP-1- derived macrophage-like cells and showed the suppression of IL-12 and S100A9 protein expression in the aortic valve. To examine the lipid-independent pleiotropic effects of pemafibrate in hVICs, conditioned-media was collected from pemafibrate-treated THP-1-derived macrophage-like cells and treated VICs with the conditioned-media (Figure 12). THP-1-derived conditioned media treatment alone increased calcification of hVICs. In contrast, pemafibrate-treated THP-1-derived conditioned-media significantly suppressed the calcification deposition in a dose-dependent manner in hVICs. In summary, pemafibrate improved aortic stenosis (AS) in a mechanical wire injury mouse model of AS. HFD accelerated AS and aortic valve calcification compared to normal diet. Pemafibrate decreased aortic valve calcification in a HFD condition with decreased inflammation and MMP activity. Pemafibrate suppressed pro-inflammatory phenotype in both in vitro and in vivo experiments. These results indicate that pemafibrate exerted the anti-calcification effects in hVICs by suppressing pro- inflammatory macrophage functions. Overall, these results show that pemafibrate exerts beneficial effects on AS and aortic valve calcification and is a treatment option for cardiovascular disease associated with calcification. OTHER EMBODIMENTS All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the same extent as if each independent publication or patent application was specifically and individually indicated to be incorporated by reference. While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations described herein following, in general, the principles described herein and including such departures from the invention that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth, and follows in the scope of the claims. Other embodiments are within the claims. PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 Some embodiments of the technology described herein can be defined according to any of the following numbered embodiments: E1. A method for treating a subject having a cardiovascular disease associated with calcification, the method comprising administering to the subject a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof. E2. The method of E1, wherein the cardiovascular disease associated with calcification is aortic stenosis. E3. The method of E2, wherein the treatment results in a decrease in calcification of the aortic valve. E4. The method of E2 or E3, wherein the treatment results in a decrease in inflammation of the aortic valve. E5. The method of any one of E2-E4, wherein the treatment results in improved open diameter narrowing of the aortic valve. E6. The method of any one of E2-E5, wherein the treatment results in improved blood flow velocity elevation in the aortic valve. E7. The method of any one of E2-E6, wherein the treatment results in a decrease in aortic valve leaflet thickness. E8. The method of E1, wherein the cardiovascular disease associated with calcification is coronary atherosclerosis. E9. The method of E8, wherein the treatment results in a decrease in calcification of the coronary arteries. E10. The method of E1, wherein the cardiovascular disease associated with calcification is peripheral artery disease (PAD). E11. The method of E10, wherein the treatment results in a decrease in calcification of the peripheral arteries. E12. The method of E1, wherein the cardiovascular disease associated with calcification is vein graft failure. E13. The method of E12, wherein the treatment results in a decrease in calcification of the vein graft. E14. The method of E1, wherein the cardiovascular disease associated with calcification is AV fistula failure. E15. The method of E14, wherein the treatment results in a decrease in calcification of the AV fistula. E16. The method of E1, wherein the cardiovascular disease associated with calcification is bicuspid aortic valves, myocardial calcification, pericardial calcification, portal vein calcification, calcific uremic arteriopathy, or Hutchinson-Gilford progeria syndrome. E17. The method of any one of E1-E16, wherein the treatment improves cardiac function or prevents heart failure. E18. The method of any one of E1-E17, wherein the subject is a human. What is claimed is:

Claims

PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 CLAIMS 1. A method for treating a subject having a cardiovascular disease associated with calcification, the method comprising administering to the subject a therapeutically effective amount of pemafibrate or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the cardiovascular disease associated with calcification is aortic stenosis.

3. The method of claim 2, wherein the treatment results in a decrease in calcification of the aortic valve.

4. The method of claim 2 or 3, wherein the treatment results in a decrease in inflammation of the aortic valve.

5. The method of claim 2, wherein the treatment results in improved open diameter narrowing of the aortic valve.

6. The method of claim 2, wherein the treatment results in improved blood flow velocity elevation in the aortic valve.

7. The method of claim 2, wherein the treatment results in a decrease in aortic valve leaflet thickness.

8. The method of claim 1, wherein the cardiovascular disease associated with calcification is coronary atherosclerosis.

9. The method of claim 8, wherein the treatment results in a decrease in calcification of the coronary arteries.

10. The method of claim 1, wherein the cardiovascular disease associated with calcification is peripheral artery disease (PAD).

11. The method of claim 10, wherein the treatment results in a decrease in calcification of the peripheral arteries.

12. The method of claim 1, wherein the cardiovascular disease associated with calcification is vein graft failure.

13. The method of claim 12, wherein the treatment results in a decrease in calcification of the vein graft.PATENT Attorney Docket No.: 51774-002WO2 MGH Ref.: 2023-261 14. The method of claim 1, wherein the cardiovascular disease associated with calcification is AV fistula failure.

15. The method of claim 14, wherein the treatment results in a decrease in calcification of the AV fistula.

16. The method of claim 1, wherein the cardiovascular disease associated with calcification is bicuspid aortic valves, myocardial calcification, pericardial calcification, portal vein calcification, calcific uremic arteriopathy, or Hutchinson-Gilford progeria syndrome.

17. The method of claim 1, wherein the treatment improves cardiac function or prevents heart failure.

18. The method of claim 1, wherein the subject is a human.

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