2-Fluoroalkyl-1,3,4-oxadiazol-5-yl-thiazoles, HDAC6 inhibitors for use in the treatment of metabolic diseases and HFPEF

Abstract

Provided herein is a method for treating or preventing metabolic disease using HDAC6 inhibitors.Also provided herein is a method for treating or preventing heart failure with preserved ejection fraction using HDAC6 inhibitors.Described herein are various HDAC6 inhibitors for use in treating or preventing metabolic disease or heart failure with preserved ejection fraction.In one aspect, described herein is a method for treating human patients by orally administering HDAC6 inhibitors, such as inhibitors of formula (I) or formula (II).

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 183,914, filed May 4, 2021, U.S. Provisional Patent Application No. 63 / 210,690, filed June 15, 2021, and U.S. Provisional Patent Application No. 63 / 210,676, filed June 15, 2021, the entire disclosures of each of which are incorporated herein by reference in their entirety.

[0002] The present disclosure relates to the treatment of metabolic disorders. The present disclosure also relates to the treatment of heart failure with preserved ejection fraction (HFpEF). [Background technology]

[0003] Metabolic diseases are a major cause of health problems, especially in countries where high-calorie diets are typical. Specifically, diet-induced obesity (DIO) affects an increasing number of people worldwide. Obesity-related metabolic diseases include diabetes and prediabetes, diabetic cardiomyopathy, metabolic syndrome (also called metabolic-related heart-related disorders), hypertension, and hypertriglyceridemia / dyslipidemia.

[0004] Heart failure with preserved ejection fraction (HFpEF) is a major health problem without effective therapy. The syndrome is increasing in prevalence and is associated with high morbidity and mortality. HFpEF, also known as diastolic heart failure, affects at least 2.5 million people annually in the United States alone. (Gazewood and Turner. Am Fam Physician. 2017 Nov 1;96(9):582-588.) Risk factors include older age, female sex, obesity, hypertension, smoking, diabetes mellitus, coronary artery disease (CAD), valvular heart disease, and atrial fibrillation. In 2013, medical costs directly attributable to heart failure totaled approximately $30 billion.

[0005] Finding effective pharmaceutical treatments for obesity and related metabolic diseases remains difficult. There is an unmet need for treatments for metabolic diseases.

[0006] Finding effective pharmaceutical treatments for HFpEF also remains difficult. The most common pharmaceutical treatment for HFpEF remains the administration of diuretics. There is an unmet need for treatment of HFpEF. The present disclosure addresses these needs. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Gazewood and Turner.Am Fam Physician.2017 Nov 1;96(9):582-588. Summary of the Invention [Means for solving the problem]

[0008] In some aspects, the disclosure relates to methods of treating or preventing a metabolic disease by administering an HDAC6 inhibitor.

[0009] In some embodiments, the disclosure provides a method of treating a metabolic disease in a subject in need thereof, the method comprising administering an HDAC6 inhibitor to the subject. In some embodiments, the disclosure provides a method of preventing a metabolic disease in a subject in need thereof, the method comprising administering an HDAC6 inhibitor to the subject. In some embodiments, the metabolic disease is an obesity-related metabolic disease, such as diet-induced obesity. In some embodiments, the metabolic disease is not diet-induced. In some embodiments, the metabolic disease is diabetes, prediabetes, diabetic cardiomyopathy, metabolic syndrome, hypertension, hypertriglyceridemia, or dyslipidemia. In some embodiments, the metabolic disease is type 2 diabetes.

[0010] In some embodiments, the disclosure provides a method of treating or preventing diabetes in a subject in need thereof, the method comprising administering an HDAC6 inhibitor. In some embodiments, the disclosure provides a method of treating or preventing diabetes mellitus in a subject in need thereof, the method comprising administering an HDAC6 inhibitor. In some embodiments, the disclosure provides a method of treating or preventing type 2 diabetes in a subject in need thereof, the method comprising administering an HDAC6 inhibitor. In some embodiments, the disclosure provides a method of treating or preventing prediabetes in a subject in need thereof, the method comprising administering an HDAC6 inhibitor. In some embodiments, the disclosure provides a method of treating or preventing diabetic cardiomyopathy in a subject in need thereof, the method comprising administering an HDAC6 inhibitor. In some embodiments, the disclosure provides a method of treating or preventing metabolic syndrome in a subject in need thereof, the method comprising administering an HDAC6 inhibitor. In some embodiments, the disclosure provides a method of treating or preventing hypertension in a subject in need thereof, the method comprising administering an HDAC6 inhibitor. In some embodiments, the disclosure provides a method of treating or preventing metabolic hypertriglyceridemia and / or dyslipidemia in a subject in need thereof, the method comprising administering an HDAC6 inhibitor. In some embodiments, the subject is obese. In other embodiments, the subject is not obese.

[0011] In some aspects, the disclosure provides a method of treating or preventing obesity in a subject in need thereof, the method comprising administering a HDAC6 inhibitor to the subject. The subject may be an obese subject or a subject at risk of obesity.

[0012] In some embodiments, the subject has or is at risk for hypertension. In some embodiments, the subject has or is at risk for diabetes (e.g., diabetes mellitus or type 2 diabetes). In some embodiments, the subject has or is at risk for diabetic cardiomyopathy. In some embodiments, the subject has or is at risk for metabolic syndrome. In some embodiments, the subject has or is at risk for hyperglyceridemia or dyslipidemia.

[0013] In some embodiments, the subject is a human. In some embodiments, the subject is at least 65 years old. In some embodiments, the subject is at least 70 years old.

[0014] In some embodiments, the disclosure provides a method of treating a metabolic disease (e.g., diabetes, such as diabetes mellitus, or metabolic syndrome) or obesity in a subject in need thereof, the method comprising administering to the subject an HDAC6 inhibitor, wherein the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative. In some embodiments, the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative according to the following formula: [ka]

[0015] In some embodiments, the disclosure provides a method of preventing a metabolic disease (e.g., diabetes, such as diabetes mellitus, or metabolic syndrome) in a subject in need thereof, the method comprising administering to the subject an HDAC6 inhibitor, wherein the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative. In some embodiments, the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative according to the following formula: [ka]

[0016] In some embodiments, the method treats or prevents at least one symptom of a metabolic disease. In some embodiments, the method improves glucose tolerance. In some embodiments, the method improves insulin resistance. In some embodiments, the method reduces glucose levels. In some embodiments, the method inhibits inflammatory genes in adipose tissue. In some embodiments, the method reduces body weight in a subject.

[0017] In another aspect, the disclosure provides an HDAC6 inhibitor for use in a method for treating a metabolic disease.

[0018] In other aspects, the disclosure provides pharmaceutical compositions for use in methods for treating metabolic diseases comprising HDAC6 inhibitors.

[0019] In other aspects, the disclosure provides kits comprising an HDAC6 inhibitor and instructions for use in a method for treating a metabolic disease.

[0020] In another aspect, the disclosure provides the use of an HDAC6 inhibitor in the treatment of a metabolic disease.

[0021] In some aspects, the disclosure relates to methods of treating or preventing heart failure with preserved ejection fraction (HFpEF) by administering an HDAC6 inhibitor.

[0022] In some embodiments, the present disclosure provides a method of treating heart failure with preserved ejection fraction (HFpEF) in a subject in need thereof, the method comprising administering an HDAC6 inhibitor to the subject. In some embodiments, the present disclosure provides a method of preventing heart failure with preserved ejection fraction (HFpEF) in a subject in need thereof, the method comprising administering an HDAC6 inhibitor to the subject.

[0023] In some aspects, the disclosure provides a method of treating or preventing cardiac fibrosis (e.g., associated with HFpEF) in a subject in need thereof, the method comprising administering an HDAC6 inhibitor to the subject.

[0024] In some aspects, the disclosure provides a method of treating or preventing diastolic dysfunction (e.g., associated with HFpEF) in a subject in need thereof, the method comprising administering an HDAC6 inhibitor to the subject.

[0025] In some embodiments, the subject has or is at risk for metabolic disease. In some embodiments, the subject has or is at risk for hypertension. In some embodiments, the subject has or is at risk for diabetes (e.g., diabetes mellitus or type 2 diabetes). In some embodiments, the subject has or is at risk for coronary artery disease (CAD). In some embodiments, the subject has or is at risk for valvular heart disease. In some embodiments, the subject has or is at risk for atrial fibrillation. In some embodiments, the subject has or is at risk for metabolic syndrome. In some embodiments, the subject is obese.

[0026] In some embodiments, the subject does not have a metabolic disease. In some embodiments, the subject does not have metabolic syndrome. In some embodiments, the subject does not have diabetes (e.g., does not have diabetes mellitus or type 2 diabetes). In some embodiments, the subject does not have hypertension. In some embodiments, the subject is not obese.

[0027] In some embodiments, the subject is a human. In some embodiments, the subject is at least 65 years old. In some embodiments, the subject is at least 70 years old.

[0028] In some embodiments, the method treats or prevents at least one symptom of HFpEF. In some embodiments, the method reduces left ventricular (LV) mass. In some embodiments, the method reduces LV wall thickness. In some embodiments, the method improves LV relaxation. In some embodiments, the method improves LV filling pressures. In some embodiments, the method prevents heart failure in a subject.

[0029] In some embodiments, the methods described herein reduce cardiac fibrosis (e.g., associated with HFpEF). In some embodiments, administration of an HDAC6 inhibitor is effective to significantly reduce cardiac fibroblast activation in cells (e.g., cell culture or in vivo). In some embodiments, administration of an HDAC6 inhibitor is effective to significantly reduce cardiac fibroblast activation in cells (e.g., cell culture or in vivo) by, for example, 10%, 20%, 30%, 40%, 50%, 60%, 80%, 90% or more (compared to before administration or in the absence of administration). In some embodiments, administration of an HDAC6 inhibitor is effective to significantly reduce the expression of one or more genes associated with fibrosis (e.g., in cells of a subject after administration of an HDAC6 inhibitor to the subject). In some embodiments, administration of an HDAC6 inhibitor is effective to reduce expression of one or more genes associated with fibrosis (e.g., in a subject's cells after administration of an HDAC6 inhibitor to the subject), for example, by 10%, 20%, 30%, 40%, 50%, 60%, 80%, 90% or more (compared to before administration or in the absence of administration).

[0030] In some embodiments, administration of HDAC6 inhibitors is effective to significantly reduce TGF-beta receptor signaling (e.g., in cells of a subject after administration of HDAC6 inhibitors to a subject). In some embodiments, administration of HDAC6 inhibitors is effective to reduce TGF-beta receptor signaling (e.g., in cells of a subject after administration of HDAC6 inhibitors to a subject), for example, by 10%, 20%, 30%, 40%, 50%, 60%, 80%, 90% or more (compared to before administration or in the absence of administration). In some embodiments, administration of HDAC6 inhibitors is effective to significantly reduce the expression of one or more genes associated with TGF-beta receptor signaling (e.g., in cells of a subject after administration of HDAC6 inhibitors to a subject). In some embodiments, administration of an HDAC6 inhibitor is effective to reduce expression of one or more genes associated with TGF-beta receptor signaling (e.g., in a subject's cells after administration of an HDAC6 inhibitor to the subject) by, for example, 10%, 20%, 30%, 40%, 50%, 60%, 80%, 90% or more (compared to before administration or in the absence of administration).

[0031] In some embodiments, the methods described herein reduce myocardial hypertrophy (eg, associated with HFpEF).

[0032] In some embodiments, the methods described herein reduce mitochondrial dysfunction. In some embodiments, administration of an HDAC6 inhibitor is effective to significantly increase the expression of genes associated with oxidative phosphorylation and / or mitochondrial complex I (e.g., in the subject's cells after administration of an HDAC6 inhibitor to the subject). In some embodiments, administration of an HDAC6 inhibitor is effective to increase the expression of genes associated with oxidative phosphorylation and / or mitochondrial complex I (e.g., in the subject's cells after administration of an HDAC6 inhibitor to the subject) by, for example, 10%, 20%, 30%, 40%, 50%, 60%, 80%, 90% or more (compared to before administration or in the absence of administration). In some embodiments, administration of an HDAC6 inhibitor is effective to increase (e.g., significantly increase) mitochondrial membrane potential in a cell (e.g., in vitro or in vivo). In some embodiments, administration of an HDAC6 inhibitor is effective to increase (e.g., significantly increase) spare respiratory capacity in a cell (e.g., cell culture or in vivo).

[0033] In some embodiments, the disclosure provides a method of treating HFpEF in a subject in need thereof, the method comprising administering an HDAC6 inhibitor, wherein the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative. In some embodiments, the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative according to the following formula: [ka]

[0034] In some embodiments, the disclosure provides a method of preventing HFpEF in a subject in need thereof, the method comprising administering an HDAC6 inhibitor, the HDAC6 inhibitor being a fluoroalkyl-oxadiazole derivative. In some embodiments, the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative according to the following formula: [ka]

[0035] In another aspect, the disclosure provides an HDAC6 inhibitor for use in a method for treating heart failure with preserved ejection fraction.

[0036] In another aspect, the disclosure provides a pharmaceutical composition for use in a method for treating heart failure with preserved ejection fraction comprising an HDAC6 inhibitor.

[0037] In other aspects, the disclosure provides a kit comprising an HDAC6 inhibitor and instructions for use in a method for treating heart failure with preserved ejection fraction.

[0038] In another aspect, the disclosure provides the use of an HDAC6 inhibitor in the treatment of heart failure with preserved ejection fraction.

[0039] In some embodiments of any of the methods described herein, administration to a subject is oral (e.g., oral administration to a human subject). In some embodiments, the treatment reduces or alleviates (i.e., is effective to reduce or alleviate) one or more symptoms or parameters of the disease being treated.

[0040] In some embodiments, the method includes selecting an HDAC6 inhibitor by performing in vitro testing for selective inhibition of HDAC6 on each member of the plurality of candidate compounds, thereby identifying a selected compound for use as an HDAC6 inhibitor.

[0041] In some embodiments, the HDAC6 inhibitor is a compound according to formula (I): [ka] where: R 1 is selected from the group consisting of: [Table 19] Ra H, halo, C 1-3 selected from the group consisting of alkyl, cycloalkyl, haloalkyl, and alkoxy; R 2 and R 3 is independently selected from the group consisting of H, halogen, alkoxy, haloalkyl, aryl, heteroaryl, alkyl, and cycloalkyl, each of which is optionally substituted, or R 2 and R 3 together with the atom to which they are attached form a cycloalkyl or heterocyclyl; R 4 and R 5 are independently H, -(SO2)R 2 , -(SO2)NR 2 R 3 , -(CO)R 2 , -(CONR 2 R 3 ), aryl, arylheteroaryl, alkylenearyl, heteroaryl, cycloalkyl, heterocyclyl, alkyl, haloalkyl, and alkoxy, each of which is optionally substituted; or R 4 and R 5 together with the atom to which they are attached form a cycloalkyl or heterocyclyl, each of which is optionally substituted; R 9 is selected from the group consisting of H, C1-C6 alkyl, haloalkyl, cycloalkyl, and heterocyclyl; X 1 are S, O, NH, and NR 6 R 6 is selected from the group consisting of C1-C6 alkyl, alkoxy, haloalkyl, cycloalkyl, and heterocyclyl; Y is CR 2 , O, N, S, SO, and SO2, and when Y is O, S, SO, or SO2, R 5 does not exist, and R 4 and R 5When taken together with the atom to which they are attached form a cycloalkyl or heterocyclyl, Y is CR 2 or N, n is selected from 0, 1, and 2.

[0042] In some embodiments, the HDAC6 inhibitor is a compound according to formula (Ik): [ka] or a pharma- ceutically acceptable salt thereof; During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted.

[0043] In some embodiments of Formula (Ik), R b is H, halogen, haloalkyl, or haloalkoxy.

[0044] In some embodiments of Formula (Ik), R 4 is optionally substituted alkyl or cycloalkyl.

[0045] In some embodiments, the HDAC6 inhibitor is a compound according to formula (Ik-1): [ka] or a pharma- ceutically acceptable salt thereof; During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted.

[0046] In some embodiments of formula (Ik-1), R b is H, halogen, haloalkyl, or haloalkoxy.

[0047] In some embodiments of formula (Ik-1), R 4 is optionally substituted alkyl or cycloalkyl.

[0048] In some embodiments of formula (Ik-1), R 4 is alkyl.

[0049] In some embodiments, the HDAC6 inhibitor is a compound according to formula (Ik-2): [ka] or a pharma- ceutically acceptable salt thereof; During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted.

[0050] In some embodiments of formula (Ik-2), R b is H, halogen, haloalkyl, or haloalkoxy.

[0051] In some embodiments of formula (Ik-2), R 4 is optionally substituted alkyl.

[0052] In some embodiments, the HDAC6 inhibitor is a compound according to formula I(y): [ka] or a pharma- ceutically acceptable salt thereof; During the ceremony, X 1 is S, R a H, halogens, and C 1-3 is selected from the group consisting of alkyl, R 1 teeth, [ka] and R 2 is selected from the group consisting of alkyl, alkoxy, and cycloalkyl, each of which is optionally substituted; R 3 is H or alkyl, R 4 is alkyl, -(SO2)R 2 , -(SO2)NR 2 R 3 , and -(CO)R 2 is selected from the group consisting of R 5 is aryl or heteroaryl, or R 4 and R 5 together with the atom to which they are attached form a heterocyclyl, each of which is optionally substituted.

[0053] In some embodiments of formula I(y), R a is H.

[0054] In some embodiments of formula I(y), R 1 teeth, [ka] It is.

[0055] In some embodiments of formula I(y), R 4 is -(SO2)R 2 It is.

[0056] In some embodiments of Formula I(y), -(SO2)R 2is -(SO2)alkyl, -(SO2)alkyleneheterocyclyl, -(SO2)haloalkyl, -(SO2)haloalkoxy, or -(SO2)cycloalkyl.

[0057] In some embodiments of formula I(y), R 5 is heteroaryl.

[0058] In some embodiments of Formula I(y), the heteroaryl is a 5-6 membered heteroaryl.

[0059] In some embodiments of Formula I(y), the 5-6 membered heteroaryl is [ka] wherein R b is halogen, alkyl, alkoxy, cycloalkyl, —CN, haloalkyl, or haloalkoxy; and m is 0 or 1.

[0060] In some embodiments of formula I(y), R b are F, Cl, -CH3, -CH2CH3, -CF3, -CHF2, -CF2CH3, -CN, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCF3, -OCHF2, -OCH2CF2H, and cyclopropyl.

[0061] In some embodiments of Formula I(y), aryl is selected from the group consisting of phenyl, 3-chlorophenyl, 3-chloro-4-fluorophenyl, 3-trifluoromethylphenyl, 3,4-difluorophenyl, and 2,6-difluorophenyl.

[0062] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0063] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0064] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0065] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0066] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0067] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0068] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0069] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0070] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0071] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0072] In some embodiments, the HDAC6 inhibitor is selected from the group consisting of: [Table 22-1] [Table 22-2]

[0073] In some embodiments, the HDAC6 inhibitor is [ka] or an analog thereof.

[0074] In some embodiments, the HDAC6 inhibitor is: [ka]

[0075] In some embodiments, the HDAC6 inhibitor is a compound of formula (II): [ka] where: n is 0 or 1, X is O, NR 4 , or CR 4 R4’ and Y is a bond, CR 2 R 3 or S(O)2, R 1 is selected from the group consisting of H, amido, carbocyclyl, heterocyclyl, aryl, and heteroaryl; R 2 and R 3 is independently selected from the group consisting of H, halogen, alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH2)-carbocyclyl, -(CH2)-heterocyclyl, -(CH2)-aryl, and -(CH2)-heteroaryl; or R 1 and R 2 together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, or R 2 and R 3 together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, R 4 and R 4’ are each independently selected from the group consisting of H, alkyl, -CO2-alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH2)-carbocyclyl, -(CH2)-heterocyclyl, -(CH2)-aryl, and -(CH2)-heteroaryl; or R 4 and R 4’ together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, Each alkyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is optionally substituted with one or more substituents independently selected from the group consisting of halogen, haloalkyl, oxo, hydroxy, alkoxy, -OCH3, -CO2CH3, -C(O)NH(OH), -CH3, morpholine, and -C(O)N-cyclopropyl.

[0076] In some embodiments, the HDAC6 inhibitor is CAY10603, tubacin, rocilinostat (ACY-1215), citarinostat (ACY-241), ACY-738, QTX-125, CKD-506, nexturastat A, tubastatin A, or HPOB. In some embodiments, the HDAC6 inhibitor is tubastatin A. In some embodiments, the HDAC6 inhibitor is ricorinostat. In some embodiments, the HDAC6 inhibitor is CAY10603. In some embodiments, the HDAC6 inhibitor is nexturastat A.

[0077] In some embodiments, the HDAC6 inhibitor is at least 100-fold selective for HDAC6 compared to all other isozymes of HDAC.

[0078] In some embodiments, the HDAC6 inhibitor reduces HDAC6 activity by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 90%, at least 95%, or at least 98%. In some embodiments, the HDAC6 inhibitor substantially eliminates HDAC6 activity. [Brief description of the drawings]

[0079] [Figure 1A](Figure 1A-L) HDAC6 inhibition with TYA-11631 improves glucose tolerance and insulin resistance in a diet-induced obese mouse model. (Figure 1A) Results of intraperitoneal glucose tolerance test (GTT) performed by injection of glucose (2 g / kg in saline) after 6 h fasting. Tail blood glucose levels (mg / dl) were measured with a glucometer before glucose administration (0 min) and 15, 30, 45, 60, and 120 min after glucose administration. Diet-induced obese (DIO) mice at 16 weeks of age developed severe glucose intolerance compared to controls. Figure 1A shows GTT results in control and DIO mice. Based on glucose AUC levels, DIO mice were randomized equally into four treatment groups to receive vehicle (n=9) or TYA-11631 at three doses of 3, 10, and 30 mg / kg (n=10 each). Control mice were also orally dosed with vehicle (n=10) or 30 mg / kg TYA-11631 (n=10) in separate doses. To evaluate the acute response of TYA-11631 on glucose metabolism, a GTT was performed 6 hours after the first dose. Figure 1B shows the GTT results before administration of TYA-11631. Figures 1C and 1D show the GTT results after a single dose of TYA-11631. A single dose of TYA-11631 at all three doses significantly reduced glucose levels. Figures 1E and 1F show the GTT results after 2 weeks of TYA-11631 treatment. Two weeks of TYA-11631 treatment resulted in a significant improvement in glucose tolerance in a dose-dependent manner. [Figure 1B] Same as above [Figure 1C] Same as above [Figure 1D] Same as above [Figure 1E] Same as above [Figure 1F] Same as above [Figure 1G]Figure 1 shows the results of an intraperitoneal insulin tolerance test (ITT) performed by injection of insulin (1 U / kg) after 6 hours of fasting. Tail blood glucose levels (mg / dl) were measured with a glucometer before insulin administration (0 min) and 15, 30, 45, 60, and 120 min after insulin administration. Four weeks of HDAC6 inhibitor (TYA-11631) treatment improved insulin resistance in DIO mice. 10 and 30 mg / kg significantly reduced glucose AUC (ITT) with comparable activity, and 3 mg / kg showed a trend toward reduction. [Figure 1H] Figure 1 shows the results of an intraperitoneal insulin tolerance test (ITT) performed by injection of insulin (1 U / kg) after 6 hours of fasting. Tail blood glucose levels (mg / dl) were measured with a glucometer before insulin administration (0 min) and 15, 30, 45, 60, and 120 min after insulin administration. Four weeks of HDAC6 inhibitor (TYA-11631) treatment improved insulin resistance in DIO mice. 10 and 30 mg / kg significantly reduced glucose AUC (ITT) with comparable activity, and 3 mg / kg showed a trend toward reduction. [Figure 1I] Figure 1 shows the effect of TYA-11631 on blood glucose in non-fasting mice. Tail blood samples were taken in the morning and measured with a glucometer. Six weeks of TYA-11631 treatment resulted in a dose-dependent reduction in non-fasting glucose, consistent with the data from the glucose tolerance test after fasting. [Figure 1J] Figure 1L shows that treatment with TYA-11631 caused a dose-dependent reduction in body weight in DIO mice. No difference in food intake was observed between groups. Of note, control mice administered TYA-11631 30 mg / kg for 6 weeks showed no change in blood glucose levels and body weight. Bars and error bars represent mean and SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. [Figure 1K]Figure 1L shows that treatment with TYA-11631 caused a dose-dependent reduction in body weight in DIO mice. No difference in food intake was observed between groups. Of note, control mice administered TYA-11631 30 mg / kg for 6 weeks showed no change in blood glucose levels and body weight. Bars and error bars represent mean and SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. [Figure 1L] Figure 1L shows that treatment with TYA-11631 caused a dose-dependent reduction in body weight in DIO mice. No difference in food intake was observed between groups. Of note, control mice administered TYA-11631 30 mg / kg for 6 weeks showed no change in blood glucose levels and body weight. Bars and error bars represent mean and SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. [Figure 2A] We show that HDAC6 inhibition with TYA-11631 inhibits inflammatory genes in adipose tissue of diet-induced obesity mouse model. White adipose tissue (epididymis) was excised from DIO mice 6 hours after a single dose of 30 mg / kg TYA-11631. Real-time q-PCR data showed that TYA-11631 significantly inhibited the upregulation of pro-inflammatory genes-IL-6 (Figure 2A), IL-10 (Figure 2B), and TGFb1 (Figure 2C) in white adipose tissue. [Figure 2B] We show that HDAC6 inhibition with TYA-11631 inhibits inflammatory genes in adipose tissue of diet-induced obesity mouse model. White adipose tissue (epididymis) was excised from DIO mice 6 hours after a single dose of 30 mg / kg TYA-11631. Real-time q-PCR data showed that TYA-11631 significantly inhibited the upregulation of pro-inflammatory genes-IL-6 (Figure 2A), IL-10 (Figure 2B), and TGFb1 (Figure 2C) in white adipose tissue. [Figure 2C]We show that HDAC6 inhibition with TYA-11631 inhibits inflammatory genes in adipose tissue of diet-induced obesity mouse model. White adipose tissue (epididymis) was excised from DIO mice 6 hours after a single dose of 30 mg / kg TYA-11631. Real-time q-PCR data showed that TYA-11631 significantly inhibited the upregulation of pro-inflammatory genes-IL-6 (Figure 2A), IL-10 (Figure 2B), and TGFb1 (Figure 2C) in white adipose tissue. [Figure 3A] Figure 3 shows TYA-11018 effects in db / db diabetic mice. Figure 3A shows that TYA-11018 caused a dose-dependent reduction in fasting glucose in db / db mice. Figure 3B shows that both 1.5 and 15 mg / kg significantly reduced blood glucose 4 hours after dosing / fasting compared to vehicle-treated db / db mice. Figure 3C shows that 15 mg / kg TYA-11018 significantly reduced fasting glucose 6 hours after dosing. Bars and error bars show mean and SEM. [Figure 3B] Figure 3 shows TYA-11018 effects in db / db diabetic mice. Figure 3A shows that TYA-11018 caused a dose-dependent reduction in fasting glucose in db / db mice. Figure 3B shows that both 1.5 and 15 mg / kg significantly reduced blood glucose 4 hours after dosing / fasting compared to vehicle-treated db / db mice. Figure 3C shows that 15 mg / kg TYA-11018 significantly reduced fasting glucose 6 hours after dosing. Bars and error bars show mean and SEM. [Figure 3C]Figure 3 shows TYA-11018 effects in db / db diabetic mice. Figure 3A shows that TYA-11018 caused a dose-dependent reduction in fasting glucose in db / db mice. Figure 3B shows that both 1.5 and 15 mg / kg significantly reduced blood glucose 4 hours after dosing / fasting compared to vehicle-treated db / db mice. Figure 3C shows that 15 mg / kg TYA-11018 significantly reduced fasting glucose 6 hours after dosing. Bars and error bars show mean and SEM. [Figure 4A] (Figure 4A-H) We show that mice fed a HFD in combination with moderate TAC develop a cardiometabolic heart failure phenotype that recapitulates the systemic and cardiovascular features of HFpEF in humans. (Figure 4A) Schematic diagram of HFpEF model induction by simultaneous metabolic and moderate pressure overload stress in wild-type mice for 12 weeks. The control group (n=5 mice) was fed a normal diet without mTAC surgery, and the HFpEF model development group (n=15 mice) was induced with HFD / mTAC. [Figure 4B] 1 shows that HFD / mTAC induced progressive weight gain and glucose intolerance compared to control mice, respectively. [Figure 4C] 1 shows that HFD / mTAC induced progressive weight gain and glucose intolerance compared to control mice, respectively. [Figure 4D] Figure 2 shows that left ventricular ejection fraction (LVEF) in mice with HFD / mTAC was preserved, as assessed by echocardiography. [Figure 4E] There was significant concentric left ventricular (LV) hypertrophy in HFD / mTAC animals, as indicated by increased LV mass ( Fig. 4E ) and LV diastolic wall thickness ( Fig. 4F ). [Figure 4F] There was significant concentric left ventricular (LV) hypertrophy in HFD / mTAC animals, as indicated by increased LV mass ( Fig. 4E ) and LV diastolic wall thickness ( Fig. 4F ). [Figure 4G]LV diastolic dysfunction accompanied by increased LV filling pressure developed in HFD / mTAC mice, as evidenced by decreased e' velocity (Fig. 4H) and increased E / e' ratio (Fig. 4G) measured by noninvasive Doppler imaging. Bars and error bars indicate mean and SEM. *P<0.05, **P<0.01, ***p<0.001 vs. control group. [Figure 4H] LV diastolic dysfunction accompanied by increased LV filling pressure developed in HFD / mTAC mice, as evidenced by decreased e' velocity (Fig. 4H) and increased E / e' ratio (Fig. 4G) measured by noninvasive Doppler imaging. Bars and error bars indicate mean and SEM. *P<0.05, **P<0.01, ***p<0.001 vs. control group. [Figure 5A] Figures 5A-5O show that oral administration of TYA-11631 improved glucose tolerance and diastolic dysfunction in HFD / mTAC mice. After the HFpEF phenotype was established, animals were randomized to receive 30 mg / kg TYA-11631 (n=8) or vehicle (n=7) orally once daily for 6 weeks. A control group of n=3 mice received vehicle. Treatment with TYA-11631 resulted in a significant improvement in glucose tolerance (Figure 5A) with no difference in body weight change compared to animals receiving vehicle (Figure 5B). Echocardiographic evaluation (see representative echocardiographically obtained M-mode tracings, Figure 5D) revealed that TYA-11631 treatment did not alter ejection fraction (Figure 5C) but significantly reduced left ventricular mass (Figure 5E) and LV wall thickness (Figure 5F). [Figure 5B] Same as above [Figure 5C] Same as above [Figure 5D] Same as above [Figure 5E] Same as above [Figure 5F] Same as above [Figure 5G]Furthermore, noninvasive Doppler imaging (representative pulsed wave Doppler (FIG. 5G) and tissue Doppler (FIG. 5J) tracings), as well as invasive catheterization analysis, demonstrated that TYA-11631 treatment maintained improvements in LV relaxation and LV filling pressures, as indicated by reduced isovolumic relaxation time prolongation (FIG. 5H), lower E / A (FIG. 5I) and E / e' ratios (FIG. 5K), improved e' velocity (FIG. 5L), and reduced end-diastolic pressure (FIG. 5M). Each of these efficacy parameters was normalized to control levels. [Figure 5H] Same as above [Figure 5I] Same as above [Figure 5J] Same as above [Figure 5K] Same as above [Figure 5L] Same as above [Figure 5M] Same as above [Figure 5N] HFD / mTAC mice treated with TYA-11631 showed a trend towards reduced heart weight (Fig. 5N) and lung weight (Fig. 5O), indicating amelioration of LV hypertrophy and pulmonary congestion, respectively, consistent with reduced filling pressures. Bars and error bars represent mean and SEM. *P<0.05, **P<0.01, ****p<0.0001. [Figure 5O] HFD / mTAC mice treated with TYA-11631 showed a trend towards reduced heart weight (Fig. 5N) and lung weight (Fig. 5O), indicating amelioration of LV hypertrophy and pulmonary congestion, respectively, consistent with reduced filling pressures. Bars and error bars represent mean and SEM. *P<0.05, **P<0.01, ****p<0.0001. [Figure 6A]TYA-11631 inhibits upregulated genes commonly associated with HFpEF disease, as shown in Figures 6A-6H. Real-time q-PCR data showed that TYA-11631 significantly inhibited the upregulation of fibrosis-related genes Postn (Figure 6A), Col1a1 (Figure 6B), Col3a1 (Figure 6C), and Col5a2 (Figure 6D); cardiac stress-related genes Nppb (Figure 6E) and Myh6 (Figure 6F); and inflammation-related genes Tnfα (Figure 6G) and Casp1 (Figure 6H) in cardiac tissue from HFD / mTAC mice, consistent with improved LV structure and cardiac function. Bars and error bars indicate mean and SEM. [Figure 6B] Same as above [Figure 6C] Same as above [Figure 6D] Same as above [Figure 6E] Same as above [Figure 6F] Same as above [Figure 6G] Same as above [Figure 6H] Same as above [Figure 7A] (Figure 7A-L) Wild-type (WT) mice on a high-fat diet with inhibition of constitutive nitric oxide synthase using N[w]-nitro-l-arginine methyl ester (HFD / L-NAME) for 8 weeks developed obesity, hypertension, and diastolic dysfunction, recapitulating the HFpEF phenotype in humans. (Figure 7A) Schematic diagram of HFpEF model induction by simultaneous metabolic and hypertensive stress in wild-type mice induced by a combination of a high-fat diet and inhibition of constitutive nitric oxide synthase using Nω-nitro-l-arginine methyl ester (L-NAME). The control group (n=7 mice) was fed a normal diet, and the HFpEF model development group (n=26 mice) was induced with HFD / L-NAME. [Figure 7B]Eight weeks of HFD / L-NAME treatment significantly induced weight gain (Figure 7B), hypertension (Figure 7C), and glucose intolerance (Figure 7D) compared with control mice. Echocardiographic evaluation revealed sustained preservation of left ventricular ejection fraction (LVEF) (Figure 7E). Significant concentric left ventricular (LV) hypertrophy was present in HFD / L-NAME mice, as shown by increased LV mass (Figure 7F) and LV wall thickness (Figure 7G) in diastole, without LV cavity dilation (Figure 7H). Furthermore, mice simultaneously exposed to HFD / L-NAME showed signs of LV diastolic dysfunction with impaired relaxation and increased LV filling pressure, as evidenced by prolonged IVRT (Figure 7I), decreased e' velocity (Figure 7J), and increased E / e' (Figure 7K), E / A (Figure 7L) ratios, measured by noninvasive Doppler imaging. Bars and error bars indicate mean and SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs. control group. [Figure 7C] Same as above [Figure 7D] Same as above [Figure 7E] Same as above [Figure 7F] Same as above [Figure 7G] Same as above [Figure 7H] Same as above [Figure 7I] Same as above [Figure 7J] Same as above [Figure 7K] Same as above [Figure 7L] Same as above [Figure 8A] (Figures 8A-8S) 9 weeks of TYA-11631 treatment improves glucose tolerance and diastolic dysfunction in the HFD / L-NAME model. (Figure 8A) Results of glucose tolerance tests (GTTs) performed after 5 weeks of treatment. Treatment with TYA-11631 significantly improved glucose tolerance in HFD / L-NAME mice, whereas control animals showed no change. [Figure 8B] Same as above [Figure 8C]Plasma insulin levels during GTT at the indicated time points (0 and 30 min after glucose injection) measured by a highly sensitive mouse insulin detection kit (catalog 80-INSMS-E01, ALPCO) are shown. TYA-11631 treatment resulted in a decrease in insulin secretion, suggesting that the improvement in glucose tolerance may be due to improved insulin action / sensitivity. [Figure 8D] 14A-14C show that treatment with TYA-11631 caused a significant reduction in body weight, but no difference in food intake, in mice fed an HFD / L-NAME. [Figure 8E] Same as above [Figure 8F] 1 shows that TYA-11631 had no effect on systolic blood pressure in HFD / L-NAME mice as measured by the non-invasive tail cuff method. [Figure 8G] Echocardiography shows that TYA-11631 treatment preserved ejection fraction (FIG. 8G), but significantly reduced left ventricular mass (FIG. 8H) and LV wall thickness (FIG. 8I). [Figure 8H] Same as above [Figure 8I] Same as above [Figure 8J] Noninvasive Doppler imaging and distal invasive catheterization analysis revealed that treatment with TYA-11631 for 9 weeks reduced isovolumic relaxation time prolongation (Figure 8J), E / A ratio (Figure 8K), and E / e' ratio (Figure 8L), increased e' velocity (Figure 8M), and reduced end-diastolic pressure (Figure 8N), indicating improved LV relaxation and LV filling pressures. Furthermore, HFD / L-NAME mice treated with TYA-11631 showed a trend toward reduced lung weight (Figure 8O), suggesting improved pulmonary congestion and consistent with reduced filling pressures. [Figure 8K] Same as above [Figure 8L] Same as above [Figure 8M] Same as above [Figure 8N] Same as above [Figure 8O] Same as above [Figure 8P]Of note, no adverse effects associated with TYA-11631 were observed. Control animals administered TYA-11631 had no changes in each of the LV structural and functional parameters, as well as ECG signals--QT, QRS, R amplitudes, and PR intervals (FIG. 8P, FIG. 8Q, FIG. 8R, and FIG. 8S, respectively), further supporting the overall favorable safety profile of the compound. Bars and error bars represent the mean and SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. [Figure 8Q] Same as above [Figure 8R] Same as above [Figure 8S] Same as above [Figure 9] Heatmap of genes selected from significantly changed functional gene sets is shown. Only genes with significant differential expression in each (GO) and WikiPathways (WP) database gene set are shown. Heatmap shows correction of key genes related to myocardium, fibrosis, and mitochondrial function. [Figure 10] Significantly altered gene sets from Gene Ontology (GO) and WikiPathways (WP) databases are shown. Selected categories were based on normalized enrichment score (NES) and FDR less than 0.25. For each condition, the left bar shows data for vehicle-treated HFpEF mice compared to healthy controls, and the right bar shows data for HFpEF mice treated with TYA-11631 compared to vehicle. TGFβ: transforming growth factor beta, PDGFR: platelet-derived growth factor receptor, OXPHOS: oxidative phosphorylation. [Figure 11A]Correlation between mitochondrial genes and diastolic function is shown. In Figures 11A-11F, the expression levels of genes related to different subunits of the mitochondrial respiratory electron transport chain (NADH:ubiquinone oxidoreductase subunits) on the x-axis are plotted against markers of diastolic function (LVPWd and MV E / E') on the y-axis. Specifically, Ndufa13 (Figure 11A), Ndufa5 (Figure 11B), Ndufs7 (Figure 11C) on the x-axis are plotted against a marker of diastolic function, LVPWd, on the y-axis. Ndufa13 (Figure 11D), Ndufa5 (Figure 11E), Ndufs7 (Figure 11F) on the x-axis are plotted against a marker of diastolic function, MV E / E', on the y-axis. For better visualization, the expression levels of genes were plotted in logarithmic scale (log2 TPM). Values ​​are Pearson correlation coefficients. Black circles indicate data from vehicle-treated healthy animals. Dark and light grey circles represent HFpEF mice treated with vehicle or TYA-11631, respectively. [Figure 11B] Same as above [Figure 11C] Same as above [Figure 11D] Same as above [Figure 11E] Same as above [Figure 11F] Same as above [Figure 11G] FIG. 11G shows an increase in mitochondrial membrane potential and FIG. 11H shows enhanced spare respiratory capacity in human iPSC-derived CMs treated with TYA-11631 (3 μM) (compared to DMSO control). Oxygen consumption rate values ​​were normalized to nuclei number. Data represent mean ± SEM. [Figure 11H] FIG. 11G shows an increase in mitochondrial membrane potential and FIG. 11H shows enhanced spare respiratory capacity in human iPSC-derived CMs treated with TYA-11631 (3 μM) (compared to DMSO control). Oxygen consumption rate values ​​were normalized to nuclei number. Data represent mean ± SEM. [Figure 12A]Shown are expression levels of fibrosis-related genes, Col1a2 (FIG. 12A), Col3a1 (FIG. 12B), Fbn1 (FIG. 12C), on the x-axis plotted against diastolic function parameters, LVPWd, on the y-axis. [Figure 12B] Shown are expression levels of fibrosis-related genes, Col1a2 (FIG. 12A), Col3a1 (FIG. 12B), Fbn1 (FIG. 12C), on the x-axis plotted against diastolic function parameters, LVPWd, on the y-axis. [Figure 12C] Shown are expression levels of fibrosis-related genes, Col1a2 (FIG. 12A), Col3a1 (FIG. 12B), Fbn1 (FIG. 12C), on the x-axis plotted against diastolic function parameters, LVPWd, on the y-axis. [Figure 12D] Figures 12D-12F show the expression levels of fibrosis-related genes, Col1a2 (Figure 12D), Col3a1 (Figure 12E), and Fbn1 (Figure 12F), on the x-axis plotted against the diastolic function parameter, IVRT, on the y-axis. [Figure 12E] Figures 12D-12F show the expression levels of fibrosis-related genes, Col1a2 (Figure 12D), Col3a1 (Figure 12E), and Fbn1 (Figure 12F), on the x-axis plotted against the diastolic function parameter, IVRT, on the y-axis. [Figure 12F] Figures 12D-12F show the expression levels of fibrosis-related genes, Col1a2 (Figure 12D), Col3a1 (Figure 12E), and Fbn1 (Figure 12F), on the x-axis plotted against the diastolic function parameter, IVRT, on the y-axis. [Figure 13A]Figure 13A shows that HDAC6 inhibitors prevent fibroblast activation from TGF-β in human cardiac fibroblasts. Figure 13A shows immunostaining of alpha-SMA (smooth muscle actin) in control human cardiac fibroblasts (first panel from the top) treated with vehicle in the presence of TGF-beta (second panel from the top) or 1 uM TYA-11631 in the presence of TGF-beta (third panel from the top). 10 ng / ml TGF-beta was used. Scale bar is 50 μM. Figure 13B shows that TYA-11631 (1 uM) effectively reduced human cardiac fibroblast activation induced by TGF-beta as measured by alpha-SMA+ cells. Alpha-SMA staining counts were determined by blinded analysis. Each point represents 9 images per well. Data represent mean ± SEM. [Figure 13B] Figure 13A shows that HDAC6 inhibitors prevent fibroblast activation from TGF-β in human cardiac fibroblasts. Figure 13A shows immunostaining of alpha-SMA (smooth muscle actin) in control human cardiac fibroblasts (first panel from the top) treated with vehicle in the presence of TGF-beta (second panel from the top) or 1 uM TYA-11631 in the presence of TGF-beta (third panel from the top). 10 ng / ml TGF-beta was used. Scale bar is 50 μM. Figure 13B shows that TYA-11631 (1 uM) effectively reduced human cardiac fibroblast activation induced by TGF-beta as measured by alpha-SMA+ cells. Alpha-SMA staining counts were determined by blinded analysis. Each point represents 9 images per well. Data represent mean ± SEM. [Figure 14A] We show that a single dose of TYA-11018 (a HDAC6-selective inhibitor with similar potency to TYA-11631) significantly improved glucose tolerance to levels similar to empagliflozin (an SGLT2-selective inhibitor, 10 mg / kg) in an established mouse HFpEF model. [Figure 14B]These results show that chronic treatment with TYA-11018 for 9 weeks reduced LV hypertrophy (FIG. 14B) and improved diastolic function, as measured by E / e' (FIG. 14C) and end-diastolic pressure LVEDP (FIG. 14D). [Figure 14C] These results show that chronic treatment with TYA-11018 for 9 weeks reduced LV hypertrophy (FIG. 14B) and improved diastolic function, as measured by E / e' (FIG. 14C) and end-diastolic pressure LVEDP (FIG. 14D). [Figure 14D] These results show that chronic treatment with TYA-11018 for 9 weeks reduced LV hypertrophy (FIG. 14B) and improved diastolic function, as measured by E / e' (FIG. 14C) and end-diastolic pressure LVEDP (FIG. 14D). [Figure 14E] Real-time q-PCR data showed that 9 weeks of TYA-11018 treatment resulted in significant inhibition of Nppb (FIG. 14E) and Col3a1 (FIG. 14F) in cardiac tissues of HFpEF mice compared to the vehicle group. Notably, TYA-11018 showed superior efficacy in inhibiting these genes compared to empagliflozin. Bars and error bars indicate the mean. [Figure 14F] Real-time q-PCR data showed that 9 weeks of TYA-11018 treatment resulted in significant inhibition of Nppb (FIG. 14E) and Col3a1 (FIG. 14F) in cardiac tissues of HFpEF mice compared to the vehicle group. Notably, TYA-11018 showed superior efficacy in inhibiting these genes compared to empagliflozin. Bars and error bars indicate the mean. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0080] overview The present disclosure generally relates to the in vivo demonstration of the effectiveness of HDAC6 inhibitors in metabolic disease.Specifically, as disclosed herein, when animal models of metabolic disease (including diet-induced obesity model and diet-independent diabetes model) are administered HDAC6 inhibitors, various physiological indicators of metabolic disease are improved.Furthermore, the efficacy of many different HDAC6 inhibitors against HDAC6 is disclosed herein.Therefore, the present disclosure provides support for the use of HDAC6 inhibitors for the treatment of metabolic disease.

[0081] In some aspects, the disclosure provides a method of treating or preventing a metabolic disease (such as any metabolic disease described herein) in a subject in need thereof, the method comprising administering a therapeutically effective amount of an HDAC6 inhibitor. In some aspects, the disclosure provides a method of treating or preventing metabolic syndrome in a subject in need thereof, the method comprising administering a therapeutically effective amount of an HDAC6 inhibitor. In some aspects, the disclosure provides a method of treating or preventing diabetes (e.g., diabetes mellitus) in a subject in need thereof, the method comprising administering a therapeutically effective amount of an HDAC6 inhibitor. In some aspects, the disclosure provides a method of treating obesity in a subject in need thereof, the method comprising administering a therapeutically effective amount of an HDAC6 inhibitor.

[0082] In some embodiments, provided herein is a method of treating or preventing a metabolic disease (e.g., a metabolic disease, e.g., diabetes or metabolic syndrome, or obesity) in a subject in need thereof, the method comprising orally administering an HDAC6 inhibitor to a human subject.

[0083] In another aspect, the present disclosure generally relates to the in vivo demonstration of the effectiveness of HDAC6 inhibitors in heart failure with preserved ejection fraction (HFpEF).Specifically, as disclosed herein, when animal models of HFpEF are administered HDAC6 inhibitors, various physiological indicators of HFpEF are improved.Furthermore, the efficacy of a number of different HDAC6 inhibitors against HDAC6 is disclosed herein.Therefore, the present disclosure provides support for the use of HDAC6 inhibitors for the treatment of HFpEF.

[0084] In some aspects, the disclosure provides a method of treating or preventing heart failure with preserved ejection fraction (HFpEF) in a subject in need thereof, the method comprising administering a therapeutically effective amount of an HDAC6 inhibitor.

[0085] In some embodiments, provided herein are methods of treating or preventing HFpEF in a subject in need thereof, the methods comprising orally administering an HDAC6 inhibitor to a human subject.

[0086] In some aspects, the disclosure provides methods of treating or preventing cardiac fibrosis in a subject in need thereof, the methods comprising administering a therapeutically effective amount of an HDAC6 inhibitor. In some embodiments, provided herein are methods of treating or preventing cardiac fibrosis associated with HFpEF in a subject in need thereof, the methods comprising administering (e.g., orally) an HDAC6 inhibitor to a subject (e.g., a human).

[0087] In some aspects, the disclosure provides a method of treating or preventing diastolic dysfunction in a subject in need thereof, the method comprising administering a therapeutically effective amount of an HDAC6 inhibitor. In some embodiments, provided herein is a method of treating or preventing diastolic dysfunction associated with HFpEF in a subject in need thereof, the method comprising administering (e.g., orally) an HDAC6 inhibitor to a subject (e.g., a human).

[0088] Advantageously, administration of selective HDAC6 inhibitors may be less toxic than pan-HDAC inhibitors.Without being bound by theory, HDAC6 inhibitors may (1) act as anti-inflammatory agents by inhibiting pro-inflammatory genes in white adipose tissue, which is the main cause of obesity-related inflammation leading to insulin resistance and metabolic dysfunction, and / or (2) improve mitochondrial activity / function.Without being bound by theory, HDAC6 inhibitors may also 1) act directly at the sarcomere level by protecting microtubules from mechanical damage, 2) improve muscle cell compliance, and / or 3) promote autophagy flux and clearance of misfolded and damaged proteins.HDAC6 inhibition may directly stabilize and protect microtubules against damage and protect Z-disks.

[0089] definition Unless otherwise indicated by the context, it is specifically contemplated that the various features of the present invention may be used in any combination. Moreover, the present disclosure also contemplates that in some embodiments, any feature or combination of features set forth herein may be excluded or omitted. For illustrative purposes, if a complex is described herein as comprising components A, B, and C, it is specifically contemplated that any of A, B, or C, or combinations thereof, alone or in any combination, may be omitted and discarded.

[0090] For example, all numerical designations, such as pH, temperature, time, concentration, and molecular weight, including ranges, are approximations that vary (+) or (-) by increments of 1.0 or 0.1, or alternatively by a variation of + / - 15%, or alternatively 10%, or alternatively 5%, or alternatively 2%. It is to be understood, although not necessarily expressly stated, that all numerical designations are preceded by the term "about". It is to be understood that such range formats are used for convenience and brevity, and should be understood to be flexible, not only including the numerical values ​​expressly specified as the limits of the range, but also to include all individual numerical values ​​or subranges subsumed within the range to the same extent as if each numerical value and subrange were expressly specified. For example, a ratio in the range of about 1 to about 200 should be understood to include not only the explicitly recited limits of about 1 to about 200, but also the individual ratios such as about 2, about 3, and about 4, and subranges such as about 10 to about 50, about 20 to about 100, etc. It is also to be understood, although not always explicitly stated, that the reagents described herein are exemplary only and that equivalents thereof are known in the art.

[0091] Also, as used herein, "and / or" refers to and includes every possible combination of one or more of the associated listed items, as well as the lack of combination when interpreted in the alternative ("or").

[0092] The term "a" or "an" may refer to one or more of that entity, i.e., it may refer to a plurality of referents. Thus, the terms "a", "an", "one or more", and "at least one" are used interchangeably herein. Furthermore, reference to an "element" by the indefinite article "a" or "an" does not exclude the possibility that more than one of the element is present, unless the context clearly requires that only one of that element is present.

[0093] Throughout this application, the term "about" is used to indicate that a value includes the inherent variation of error for the device or method used to determine the value or variation present between samples being measured. Unless otherwise stated or otherwise clear from the context, the term "about" means within a range of 10% above or below the reported numerical value (except where such number would be greater than 100% or less than 0% of the possible values). When used in conjunction with a range or series of values, the term "about" applies to the endpoints of the range or each of the values ​​recited in the series, unless otherwise indicated. As used in this application, the terms "about" and "approximately" are used as equivalents.

[0094] As used herein, the term "HDAC6" refers to the enzyme encoded by the HDAC6 gene in humans.

[0095] As used herein, the term "HDAC6 inhibitor" refers to a compound that inhibits at least one enzymatic activity of HDAC6.

[0096] The HDAC6 inhibitor may be a "selective" HDAC6 inhibitor. The term "selective" as used herein refers to selectivity over other HDACs known in the art as "isoenzymes". In some embodiments, the selectivity ratio of HDAC6 to HDAC1 is about 5 to about 30,0000, for example, about 5, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 1000, about 2000, about 3000, about 4000, about 5000, about 6000, about 7000, about 8000, about 9000, about 10,000, about 15,000, about 20,000, about 25,000, about 30,000, including all values ​​and ranges therebetween.

[0097] For example, HDAC6 inhibitors can be at least 100 times more selective for HDAC6 than all other isozymes of HDAC.In some cases, selectivity can be determined by reference to another HDAC inhibitor, such as a pan-HDAC inhibitor, which is an inhibitor that inhibits HDAC6 as well as HDACs other than HDAC6.Givinostat is an example of a pan-HDAC6 inhibitor.In some embodiments, selective HDAC6 inhibitors inhibit HDACs other than HDAC6 with at least 100 times less effectiveness than givinostat.

[0098] "Alkyl" or "alkyl group" refers to a fully saturated straight or branched hydrocarbon chain having from 1 to 12 carbon atoms and attached to the remainder of the molecule by a single bond. Alkyl containing any number of carbon atoms from 1 to 12 is included. Alkyl containing up to 12 carbon atoms is any of the groups C1-C 12 Alkyl, containing up to 10 carbon atoms, is C1-C 10 Alkyl, an alkyl containing up to 6 carbon atoms is C1-C6 alkyl, and an alkyl containing up to 5 carbon atoms is C1-C5 alkyl. C1-C5 alkyl includes C5 alkyl, C4 alkyl, C3 alkyl, C2 alkyl, and C1 alkyl (i.e., methyl). C1-C6 alkyl includes all of the moieties listed above for C1-C5 alkyl, but also includes C6 alkyl. C1-C 10 Alkyl includes all of the moieties described above for C1-C5 alkyl and C1-C6 alkyl, but also includes C7, C8, C9 and C 10 Also includes alkyl. Similarly, C1-C 12 Alkyl includes all of the above moieties, but C 11 and C 12 Includes alkyl. C1-C 12Non-limiting examples of alkyl include methyl, ethyl, n-propyl, i-propyl, sec-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, t-amyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, and n-dodecyl. Unless stated otherwise in the specification, an alkyl group can be optionally substituted.

[0099] "Alkylene" or "alkylene chain" refers to a fully saturated, straight or branched divalent hydrocarbon chain radical having 1 to 12 carbon atoms. 12 Non-limiting examples of alkylene include methylene, ethylene, propylene, n-butylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group (e.g., those described herein) through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise in the specification, an alkylene chain can be optionally substituted.

[0100] "Alkenyl" or "alkenyl group" refers to a straight or branched hydrocarbon chain having from 2 to 12 carbon atoms and having one or more carbon-carbon double bonds. Each alkenyl group is attached to the remainder of the molecule by a single bond. Alkenyl groups containing any number of carbon atoms from 2 to 12 are included. Alkenyl groups containing up to 12 carbon atoms include C2-C 12 Alkenyl containing up to 10 carbon atoms is C2-C 10 Alkenyl, an alkenyl group containing up to 6 carbon atoms is C2-C6 alkenyl, and an alkenyl containing up to 5 carbon atoms is C2-C5 alkenyl. C2-C5 alkenyl includes C5 alkenyl, C4 alkenyl, C3 alkenyl, and C2 alkenyl. C2-C6 alkenyl includes all of the moieties listed above for C2-C5 alkenyl, but also includes C6 alkenyl. C2-C 10Alkenyl includes all of the moieties listed above for C2-C5 alkenyl and C2-C6 alkenyl, but also includes C7, C8, C9 and C 10 Also includes alkenyl. Similarly, C2-C 12 Alkenyl includes all of the above moieties, except that C 11 and C 12 Includes alkenyl. C2-C 12 Non-limiting examples of alkenyl include ethenyl (vinyl), 1-propenyl, 2-propenyl (allyl), iso-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl, 1-octenyl, 2-octenyl, 3-octenyl, 4-octenyl, 5-octenyl, 6-octenyl, 7-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, Examples include 4-nonenyl, 5-nonenyl, 6-nonenyl, 7-nonenyl, 8-nonenyl, 1-decenyl, 2-decenyl, 3-decenyl, 4-decenyl, 5-decenyl, 6-decenyl, 7-decenyl, 8-decenyl, 9-decenyl, 1-undecenyl, 2-undecenyl, 3-undecenyl, 4-undecenyl, 5-undecenyl, 6-undecenyl, 7-undecenyl, 8-undecenyl, 9-undecenyl, 10-undecenyl, 1-dodecenyl, 2-dodecenyl, 3-dodecenyl, 4-dodecenyl, 5-dodecenyl, 6-dodecenyl, 7-dodecenyl, 8-dodecenyl, 9-dodecenyl, 10-dodecenyl, and 11-dodecenyl. Unless stated otherwise in the specification, an alkyl group can be optionally substituted.

[0101] "Alkenylene" or "alkenylene chain" refers to an unsaturated, linear or branched divalent hydrocarbon chain radical having one or more olefins and having 2 to 12 carbon atoms. C2-C 12Non-limiting examples of alkenylene include ethenylene, propenylene, n-butenylene, and the like. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group (e.g., those described herein) through a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise in the specification, an alkenylene chain can be optionally substituted.

[0102] "Alkynyl" or "alkynyl group" refers to a straight or branched hydrocarbon chain having from 2 to 12 carbon atoms and having one or more carbon-carbon triple bonds. Each alkynyl group is attached to the remainder of the molecule by a single bond. Alkynyl groups containing any number of carbon atoms from 2 to 12 are included. Alkynyl groups containing up to 12 carbon atoms include those having C2-C 12 Alkynyl containing up to 10 carbon atoms is C2-C 10 An alkynyl group containing up to 6 carbon atoms is C2-C6 alkynyl, and an alkynyl containing up to 5 carbon atoms is C2-C5 alkynyl. C2-C5 alkynyl includes C5 alkynyl, C4 alkynyl, C3 alkynyl, and C2 alkynyl. C2-C6 alkynyl includes all of the moieties listed above for C2-C5 alkynyl, but also includes C6 alkynyl. C2-C 10 Alkynyl includes all of the moieties listed above for C2-C5 alkynyl and C2-C6 alkynyl, but also includes C7, C8, C9 and C 10 Alkynyl is also included. Similarly, C2-C 12 Alkynyl includes all of the above moieties, except that C 11 and C 12 Alkynyl is also included. C2-C 12 Non-limiting examples of alkenyls include ethynyl, propynyl, butynyl, pentynyl, etc. Unless stated otherwise in the specification, an alkyl group can be optionally substituted.

[0103] "Alkynylene" or "alkynylene chain" refers to an unsaturated, linear or branched divalent hydrocarbon chain radical having one or more alkynes and having 2 to 12 carbon atoms. 12 Non-limiting examples of alkynylene include ethynylene, propynylene, n-butynylene, and the like. The alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group (e.g., as described herein) through a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to the radical group may be through any two carbons in the chain having suitable valences. Unless stated otherwise in this specification, the alkynylene chain can be optionally substituted.

[0104] "Alkoxy" means a group of the formula -OR a In the formula, R a is an alkyl, alkenyl, or alkynyl, as defined above, containing 1 to 12 carbon atoms. Unless stated otherwise in the specification, an alkoxy group can be optionally substituted.

[0105] "Aryl" refers to a hydrocarbon ring system containing hydrogen, 6-18 carbon atoms, and at least one aromatic ring, attached to the remainder of the molecule by a single bond. For purposes of this disclosure, an aryl may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems. Aryls include, but are not limited to, aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unless otherwise stated in this specification, an "aryl" may be optionally substituted.

[0106] "Carbocyclyl", "carbocyclic ring", or "carbocycle" refers to a ring structure in which the atoms forming the ring are each carbon and are attached to the remainder of the molecule by a single bond. Carbocyclic rings can contain from 3 to 20 carbon atoms in the ring. Carbocyclic rings include aryl and cycloalkyl, cycloalkenyl, and cycloalkynyl as defined herein. Unless stated otherwise in the specification, a carbocyclyl group can be optionally substituted.

[0107] A "carbocyclylalkyl" is a group of the formula -R b -R d where R b is an alkylene, alkenylene, or alkynylene group as defined above; R d is a carbocyclyl radical, as defined above. Unless stated otherwise in the specification, a carbocyclylalkyl group may be optionally substituted.

[0108] "Cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic fully saturated hydrocarbon (which may include fused or bridged ring systems) composed solely of carbon and hydrogen atoms, having 3 to 20 carbon atoms (e.g., having 3 to 10 carbon atoms) and attached to the remainder of the molecule by a single bond. Monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyls include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise stated in the specification, cycloalkyl groups can be optionally substituted.

[0109] "Cycloalkenyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon (which may include fused or bridged ring systems) composed solely of carbon and hydrogen atoms and having one or more carbon-carbon double bonds, having 3 to 20 carbon atoms, preferably 3 to 10 carbon atoms, and attached to the remainder of the molecule by a single bond. Monocyclic cycloalkenyls include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, cycloctenyl, and the like. Polycyclic cycloalkenyls include, for example, bicyclo[2.2.1]hept-2-enyl, and the like. Unless otherwise stated in the specification, cycloalkenyl groups can be optionally substituted.

[0110] "Cycloalkynyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon (which may include fused or bridged ring systems) composed solely of carbon and hydrogen atoms, having one or more carbon-carbon triple bonds, having 3 to 20 carbon atoms, preferably 3 to 10 carbon atoms, and attached to the remainder of the molecule by a single bond. Monocyclic cycloalkynyls include, for example, cycloheptynyl, cyclooctynyl, and the like. Unless otherwise stated in the specification, cycloalkynyl groups can be optionally substituted.

[0111] "Haloalkyl" refers to an alkyl, as defined above, substituted by one or more halo radicals, such as, for example, trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. Unless stated otherwise in the specification, a haloalkyl group may be optionally substituted.

[0112] "Heterocyclyl", "heterocyclic ring", or "heterocycle" refers to a stable saturated, unsaturated, or aromatic 3-20 membered ring, consisting of 2-19 carbon atoms and 1-6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, which is attached to the remainder of the molecule by a single bond. Heterocyclyl or heterocyclic ring includes heteroaryl, heterocyclylalkyl, heterocyclylalkenyl, and heterocyclylalkynyl. Unless otherwise stated in the specification, a heterocyclyl may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems, the nitrogen, carbon, or sulfur atoms in a heterocyclyl may be optionally oxidized, the nitrogen atom may be optionally quaternized, and the heterocyclyl may be partially or fully saturated. Examples of such heterocyclyls include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise in the specification, heterocyclyl groups may be optionally substituted.

[0113] "Heteroaryl" refers to a 5-20 membered ring system containing a hydrogen atom, 1-19 carbon atoms, 1-6 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, at least one aromatic ring, and attached to the remainder of the molecule by a single bond. For purposes of this disclosure, an aryl may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems, and the nitrogen, carbon, or sulfur atoms in a heteroaryl may be optionally oxidized, and the nitrogen atom may be optionally quaternized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, isothiophenyl ... Examples of such aryl groups include oxazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Unless stated otherwise in the specification, a heteroaryl group may be optionally substituted.

[0114] "Heterocyclylalkyl" refers to a group of the formula -R b -R e where R b is an alkylene, alkenylene, or alkynylene group as defined above; R e is a heterocyclyl radical as defined above. Unless stated otherwise in the specification, a heterocycloalkylalkyl group may be optionally substituted.

[0115] The term "substituted," as used herein, means any of the groups described herein (e.g., alkyl, alkenyl, alkynyl, alkoxy, aryl, aralkyl, carbocyclyl, cycloalkyl, cycloalkenyl, cycloalkynyl, haloalkyl, heterocyclyl, and / or heteroaryl) in which at least one hydrogen atom has been replaced by a bond to a non-hydrogen atom, such as, but not limited to, a halogen atom, such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxyl, alkoxy, and ester groups; a sulfur atom in groups such as thiol, thioalkyl, sulfone, sulfonyl, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; a silicon atom in groups such as trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, and triarylsilyl groups; and other heteroatoms in various other groups. "Substituted" also refers to any of the above groups in which one or more hydrogen atoms have been replaced by a higher order bond (e.g., a double or triple bond) to a heteroatom, such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and nitrogen in groups such as imines, oximes, hydrazones, and nitriles. For example, "substituted" refers to any group in which one or more hydrogen atoms have been replaced by a bond such as -NR g R h , -NR g C(=O)R h , -NR g C(=O)NRg R h , -NR g C(=O)OR h , -NR g SO2R h , -OC(=O)NR g R h , -OR g , -SR g , -SOR g , -SO2R g , -OSO2R g , -SO2OR g , =NSO2R g , and -SO2NR g R h "Substituted" also refers to any of the above groups in which one or more hydrogen atoms have been replaced by -C(=O)R. g , -C(=O)OR g , -C(=O)NR g R h , -CH2SO2R g , -CH2SO2NR g R h In the above, R g and R hare the same or different and independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and / or heteroarylalkyl. "Substituted" further refers to any of the above groups wherein one or more hydrogen atoms are replaced by a bond to an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and / or heteroarylalkyl group. In addition, each of the foregoing substituents may be optionally substituted with one or more of the above-described substituents.

[0116] As used herein, the symbols [ka] (hereinafter may be referred to as a "point of attachment bond") refers to a bond that is a point of attachment between two chemical entities, one of which is depicted as being attached to the point of attachment bond and the other of which is not depicted as being attached to the point of attachment bond. For example, [ka] indicates that a chemical entity "XY" is attached to another chemical entity via an attachment point bond. Furthermore, specific attachment points to undepicted chemical entities can be identified by inference. For example, R 3 Is H or [ka] The compound CH3-R 3 is R3 If is "XY", the attachment point bond is R 3 is assumed to be the same bond depicted as being attached to CH3.

[0117] As used herein, the term "treating" refers to acting on a disease, disorder, or condition with an agent to reduce or alleviate the harmful or any other undesirable effects of the disease, disorder, condition, and / or symptoms thereof.

[0118] As used herein, the term "preventing" refers to reducing the incidence or risk of developing or delaying the onset of adverse or any other undesirable effects of a disease, disorder, condition, and / or symptom.

[0119] "Administration", "administering" and the like refer to administration to a subject by a medical professional or self-administration by a subject, as well as indirect administration, which may be the act of prescribing a composition of the present invention. Typically, an effective amount is administered, which amount may be determined by one of skill in the art. Any method of administration may be used. Administration to a subject may be accomplished, for example, by oral administration in liquid or solid form, e.g., capsule or tablet form, intravascular injection, intramyocardial delivery, or other suitable administration form.

[0120] As used herein, the term "effective amount" refers to an amount that is sufficient to induce a desired physiological outcome (e.g., increased cardiac function, decreased mortality or reduced risk / incidence of hospitalization, increased exercise capacity, or reduced expression of one or more biomarkers associated with heart failure, such as BNP). An effective amount can be administered in one or more administrations, applications, or dosages. Such delivery depends on many variables, including the period for which the individual dosage units are used, the bioavailability of the composition, the route of administration, and the like. However, it is understood that the specific amount of the composition for any particular subject will depend on a variety of factors, including the activity of the particular agent used, the age, weight, general health, sex, and diet of the subject, the time of administration, the rate of excretion, the combination of the composition, the severity of the particular disease being treated, and the form of administration.

[0121] As used herein, the term "subject" or "patient" refers to any animal, such as a domestic animal, a zoo animal, or a human. A "subject" or "patient" may be a mammal, such as a dog, a cat, a horse, a livestock animal, a zoo animal, or a human. A subject or patient may also be any domestic animal, such as a bird, a pet, or a farm animal. Specific examples of "subject" and "patient" include, but are not limited to, individuals with metabolic disease (e.g., obesity) and individuals with metabolic disease-related characteristics or symptoms. Specific examples of "subject" and "patient" also include, but are not limited to, individuals with cardiac disease or disorder and individuals with cardiac disorder-related characteristics or symptoms.

[0122] The phrase "pharmacologically acceptable" as used herein refers to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0123] The term "pharmaceutically acceptable salts" includes those obtained by reacting an active compound that functions as a base with an inorganic or organic acid to form a salt, including, for example, salts of hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, camphorsulfonic acid, oxalic acid, maleic acid, succinic acid, citric acid, formic acid, hydrobromic acid, benzoic acid, tartaric acid, fumaric acid, salicylic acid, mandelic acid, carbonic acid, etc. One of ordinary skill in the art will further recognize that acid addition salts can be prepared by reacting a compound with the appropriate inorganic or organic acid via any of a number of known methods.

[0124] As used herein, the term "ameliorate" refers to increasing the level of a biochemical or physiological parameter to the level observed in a subject prior to the onset of a disease or condition, or to the level observed in a subject not having the disease or condition.

[0125] As used herein, the term "reduce" refers to decreasing the level of a biochemical or physiological parameter.

[0126] As used herein, the term "metabolic disease" refers to conditions caused by either excessive intake of nutrients or the body's inability to properly metabolize nutrients. Metabolic diseases include, but are not limited to, obesity. It is a cluster of conditions that occur together that are related to heart disease, type 2 diabetes, and stroke. These conditions include high blood pressure, hyperglycemia, excess body fat, and abnormal cholesterol or triglyceride levels.

[0127] The term "obesity" refers to the condition of having too much body fat. Obesity can increase the risk of diabetes, heart disease, stroke, and arthritis. A body weight higher than what is considered healthy for a given height is described as overweight or obesity. Body mass index (BMI) is a screening tool for overweight and obesity. BMI is a person's weight in kilograms divided by the square of their height in meters. BMI calculators are available at www.cdc.gov at obesity / adult / defining.html. In various embodiments, the subject may have a BMI of 25, 30, 35, 40 or more, such as a BMI of 25-30, 25-30, or 25-40. In some cases, the subject may have severe obesity (also known as class 3 obesity), defined as a BMI of 40 or more. Obesity is a chronic disease and a risk factor for other diseases, such as heart disease, high blood pressure, stroke, and diabetes.

[0128] In some embodiments, the methods of the present disclosure reduce, prevent, or ameliorate one or more symptoms of metabolic disease, including glucose intolerance, insulin resistance, high glucose levels, and inflammation in adipose tissue.

[0129] As used herein, the term "heart failure" refers to a condition in which the heart is unable to pump enough blood to meet the body's needs.

[0130] "Heart failure (HF)" is a complex clinical syndrome that can result from any structural or functional cardiovascular disorder that makes systemic perfusion inadequate to meet the metabolic demands of the body without excessively increasing left ventricular filling pressure. It is characterized by certain symptoms such as dyspnea and fatigue, as well as signs such as fluid retention. As used herein, "chronic heart failure" or "congestive heart failure" or "CHF" interchangeably refer to ongoing or persistent forms of heart failure. Common risk factors for CHF include advanced age, diabetes, hypertension, and being overweight. CHF is broadly classified according to the contractile function of the left ventricle as HF with reduced or preserved ejection fraction (HFrEF and HFpEF). The term "heart failure" does not mean that the heart has stopped or is not functioning completely, but rather that it is weaker than normal in healthy people. In some cases, the condition may be mild, causing symptoms that may only be noticeable during exercise, while in other cases, the condition may be more severe, causing potentially life-threatening symptoms even at rest. The most common symptoms of chronic heart failure include shortness of breath, fatigue, leg and ankle swelling, chest pain, and coughing. In some embodiments, the methods of the present disclosure reduce, prevent, or ameliorate one or more symptoms of CHF (e.g., HFpEF) in a subject suffering from or at risk of CHF (e.g., HFpEF). In some embodiments, the present disclosure provides methods of treating CHF and conditions that may lead to CHF.

[0131] As used herein, "acute heart failure" or "decompensated heart failure" interchangeably refer to a syndrome of worsening signs and symptoms reflecting the inability of the heart to pump blood at normal filling pressures at a rate equal to the body's needs. AHF typically develops gradually over days to weeks, then decompensates and requires emergency or critical care depending on the severity of these signs or symptoms. AHF can be the result of a primary impairment in cardiac systolic or diastolic function, or abnormal venous or arterial vasoconstriction, but generally represents an interaction of multiple factors, including volume overload. The majority of patients with AHF have decompensated chronic heart failure (CHF), and as a result, much of the discussion of the pathophysiology, presentation, and diagnosis of CHF is directly relevant to understanding AHF. In other cases, AHF results from cardiac injury or events that impair cardiac function, such as acute myocardial infarction, severe hypertension, heart valve damage, arrhythmias, cardiac inflammation or infection, toxins, and medications. In some embodiments, the methods of the present disclosure reduce, prevent, or ameliorate one or more symptoms of AHF in a subject suffering from or at risk for AHF. In some embodiments, the present disclosure provides methods of treating AHF and conditions that may result in AHF. AHF may be the result of ischemia associated with myocardial infarction.

[0132] In some embodiments, the disclosed methods reduce, prevent, or ameliorate one or more symptoms of heart failure in a subject suffering from or at risk for heart failure associated with HFpEF. The terms "heart failure with preserved ejection fraction" or "diastolic heart failure" are used interchangeably and generally refer to a form of heart failure characterized by signs and symptoms of heart failure and a left ventricular ejection fraction (LVEF) of greater than 50%. The term may also encompass heart failure associated with moderate reductions in LVEF (40%-49%). In some embodiments, HFpEF includes HFpEF associated with CHF. In some embodiments, HFpEF includes HFpEF associated with AHF.

[0133] HFpEF is more common among older patients and women. Typical symptoms include fatigue, weakness, dyspnea, orthopnea, paroxysmal nocturnal dyspnea, and edema. Signs of HFpEF (e.g., HFpEF associated with CHF) may include an S3 heart sound, displaced apical beat, and jugular venous distention. Echocardiographic findings of normal ejection fraction with diastolic dysfunction support the diagnosis. Measurement of natriuretic peptides (e.g., BNP or NT-proBNP) is useful in the evaluation of patients with suspected heart failure with preserved ejection fraction in the outpatient setting.

[0134] As used herein, the term "detrimental mutation" refers to a mutation that reduces the function of a gene. Detrimental mutations can include missense mutations, deletions or insertions in coding regions, non-coding mutations that affect gene expression or gene splicing, or others. Detrimental mutations include partial or complete deletions of a gene. As used herein, the term can refer to homozygous or heterozygous mutations in a gene, where the mutation has an phenotypic effect in carriers.

[0135] As used herein, the term "left ventricular internal diameter diastolic" or "LVIDd" refers to the size of the left ventricle during diastole.

[0136] As used herein, the term "left ventricular systolic dimensions" or "LVIDs" refers to the size of the left ventricle during systole.

[0137] As used herein, the term "left ventricular mass" refers to the weight of the left ventricle.

[0138] As used herein, the term "ejection fraction" means the amount of blood pushed out of the left ventricle with each contraction, expressed as a percentage of the total amount of blood in the left ventricle.

[0139] The detailed description of the present disclosure is divided into various sections for the convenience of the reader only, and the disclosure found in any section may be combined with the disclosure of another section. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. Although any methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present invention, the preferred methods and materials are now described.

[0140] Treatment or prevention of metabolic diseases Provided herein are methods of treating or preventing metabolic diseases with HDAC6 inhibitors.

[0141] Treatment or prevention of heart failure with preserved ejection fraction (HFpEF) Also provided herein are methods of treating or preventing heart failure with preserved ejection fraction (HFpEF) using HDAC6 inhibitors.

[0142] HDAC6 inhibitors Histone deacetylases ("HDACs") are a class of enzymes that have deacetylase activity with a wide range of genomic and non-genomic substrates. There are 11 zinc-dependent HDAC enzymes that have been classified based on sequence identity and catalytic activity (Haberland et al., 2009).

[0143] Histone deacetylase inhibitors have been described as therapeutic agents in oncology (Yoon and Eom, 2016), neurodegenerative (Butler et al., 2010) autoimmune diseases (Choi et al., 2018), chemotherapy-induced peripheral neuropathy (Krukowski et al., 2017), and cardiac indications (Zhang et al., 2002). Given the role of nuclear HDACs in regulating gene transcription, inhibition of these target classes is known to have pleiotropic effects in various cell types, most notably resulting in cytotoxicity. Thus, limiting the toxicity of pan-HDAC inhibitors represents a major obstacle in the widespread utilization of this compound class. Furthermore, significant adverse effects of pan-HDAC inhibitors (e.g., SAHA and Panabinostat) including fatigue, nausea, diarrhea, and thrombocytopenia have been observed in the clinic (Subramanian et al., 2010).

[0144] In the cardiac indication setting, most studies have utilized pan-HDAC inhibitors (e.g., SAHA, TSA, and givinostat) to treat pressure-overload rodent models with systolic dysfunction, including transverse aortic constriction (TAC) (Cao et al., 2011), hypertension in Dahl salt-sensitive rats (Jeong et al., 2018), and myocardial infarction (Nagata et al., 2019). Additionally, HDAC6-selective inhibitors have been used to improve LV systolic dysfunction in pressure-overload rodent modes (Demos-Davies et al., 2014) and provide protection against proteotoxicity in transgenic cardiomyopathy mouse models (McLendon et al., 2014). However, these experiments in pressure-overload rodent models, all of which were accompanied by reduced ejection fraction and exhibited a HFrEF phenotype, are not predictive of treating HFpEF. Pressure overload mediated by conventional transverse aortic constriction induces cardiac hypertrophy and subsequently heart failure with reduced ejection fraction (HFrEF) rather than HFpEF (Mohammadi et al. 2021). HFpEF is distinct from HFrEF. The two forms of heart failure are fundamentally different in their pathophysiological mechanisms and the extent of myocardial loss / dysfunction, patterns of remodeling at the luminal and ultrastructural levels, and response to therapeutic interventions (Schiattarella et al., 2020).

[0145] HDAC6 belongs to the class IIb enzymes and contains two catalytic domains, a ubiquitin-binding domain and a cytoplasmic retention domain (Haberland et al., 2009). HDAC6 is primarily a cytoplasmic enzyme whose best-characterized substrates include tubulin, HSP90 and cortactin (Brindisi et al., 2020).

[0146] Pharmacological inhibition of HDAC6 blocks its deacetylase activity, resulting in hyperacetylation of its substrates, particularly tubulin (Hubbert et al., 2002).

[0147] HDAC6-selective inhibitors are known to have reduced cytotoxicity due to the cytoplasmic nature of HDAC6 substrates and reduced effects on nuclear targets (including H3K9 and c-MYC) and global transcription ( Nebbioso et al., 2017 ).

[0148] Hydroxamic acids are zinc chelators and have been widely used in the development of pan- and HDAC-selective inhibitors. However, most hydroxamic acid-based HDAC inhibitors lack the desired selectivity or exhibit poor pharmacokinetic profiles and poor bioavailability (Butler et al., 2010; Santo et al., 2012).

[0149] A variety of selective HDAC6 are known in the art.In addition, it is common to use known methods to screen compounds to identify further selective HDAC6 inhibitors.Specifically, taking into account known HDAC6 inhibitors, those skilled in the art can identify which analogues of compounds have selective HDAC6 activity.

[0150] In some embodiments, the HDAC6 inhibitor is a gene silencing agent, such as an RNA silencing agent (e.g., siRNA).In some embodiments, the HDAC6 inhibitor is not a gene silencing agent.In some embodiments, the HDAC6 inhibitor is a small molecule HDAC6 inhibitor.

[0151] Known HDAC6 inhibitors In some embodiments, the HDAC6 inhibitor is CAY10603, tubacin, rosilinostat (ACY-1215), citalinostat (ACY-241), ACY-738, QTX-125, CKD-506, nextulastat A, tubastatin A, or HPOB (listed in Table 1), or an analog thereof. [Table 1]

[0152] More examples of HDAC6 inhibitors, U.S. Patent Publication No. 8227516 (B2), No. 2010 / 0292169 (A1), No. 200 No. 7 / 0207950(A1), same as No. 8222423(B2), same as No. 2010 / 0093824(A1), same as No. 2010 / 02167 96(A1), same as No. 8673911(B2), same as No. 8217076(B2), same as No. 8440716(B2), same as No. 2011 / 0195432(A1), same as No. 8624040(B2), same as No. 9096518(B2), same as No. 8431538(B2), same as No. 2012 / 0258993(A1), same as No. 8546588(B2), same as No. 8513421(B2), same as No. 2014 / 0031368(A1), same as No. 2012 / 0015943(A1), same as No. 2012 / 0015942(A1), same as No. 2014 / 0243335(A1), same as No. 2013 / 0225543(A1), 8471026(B2), 9238028(B2), 8765773(B2), USRE47009(E1), 2014 / 0294856(A1), 9512083(B2), 9670193(B2 ), No. 9345905(B2), No. 9409858(B2), No. 9663825(B2), No. 2015 / 0119327(A1), No. 2015 / 0250786(A1), No. 10041046(B2), No. 9586973(B2), No. 201 6 / 0069887(A1), same as No. 2014 / 0357512(A1), same as No. 9751832(B2), same as No. 2016 / 0228434(A1), same as No. 2015 / 0105358(A1), same as No. 10660890(B2), same as No. 2016 / 0271083(A1) , same as No. 2015 / 0176076(A1), same as No. 2020 / 0405716(A1), same as No. 9890136(B2), same as No. 10287255(B2), same as No. 2017 / 0173083(A1), same as No. 10016421(B2), same as No. 9987258(B2), same as No. No. 10568854 (B2), No. 10106540 ​​(B2), No. 10266489 (B2), No. 9993459 (B2), No. 10183934 (B2), No. 10494354 (B2), No. 10494353 (B2), No. 10112915 (B2),Same No. 10377726 (B2), No. 10829462 (B2), No. 10829461 (B2), No. 2021 / 0009539 (A1), No. 2021 / 0009538 (A1), No. 10239845 (B2) ) No. 10472337 (B2), No. 10479772 (B2), No. 10464911 (B2), No. 10584117 (B2), No. 10538498 (B2), No. 10011611 (B2), No. 1 0494355 (B2), 10040769 (B2), 10858323 (B2), 10654814 (B2), 2019 / 0209559 (A1), 2019 / 0185462 (A1), No. 2019 / 0192521(A1), No. 2019 / 0321361(A1), No. 2020 / 0046698(A1), No. 2019 / 0262337(A1), No. 2019 / 0282573(A1), No. 2 019 / 0282574(A1), 2020 / 0071288(A1), 10745389(B2), 10357493(B2), 2020 / 0171028(A1), 2020 / 00547 73(A1), 2020 / 0308174(A1), 2020 / 0155549(A1), 10435399(B2), 2020 / 0216563(A1), 2019 / 0216751(A1) Nos. 2020 / 0339569(A1), 2021 / 0078963(A1), 2021 / 0077487(A1), 2019 / 0270733(A1), 2019 / 0270744(A1), 2020 / 0022966(A1), and 2021 / 0094944(A1), which are incorporated herein for purposes of identifying HDAC6 inhibitors that may be used in the methods disclosed herein. In some embodiments, the HDAC6 inhibitor is TYA-631 or an analog thereof.

[0153] Fluoroalkyl-oxadiazole derivatives In some embodiments, the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative.Exemplary fluoroalkyl-oxadiazole derivatives that can be used as HDAC6 inhibitors include those described herein and those disclosed in International Patent Application No. PCT / US2020 / 066439, published as WO2021 / 127643(A1), the contents of which are incorporated herein by reference in their entirety.PCT / US2020 / 066439, published as WO2021 / 127643(A1), also describes the synthesis method of such compounds, which are specifically incorporated herein by reference.

[0154] In some embodiments, the HDAC6 inhibitor is a compound of formula (I): [ka] where: R 1 is selected from the group consisting of: [Table 20] R a H, halo, C 1-3 selected from the group consisting of alkyl, cycloalkyl, haloalkyl, and alkoxy; R 2 and R 3 is independently selected from the group consisting of H, halogen, alkoxy, haloalkyl, aryl, heteroaryl, alkyl, and cycloalkyl, each of which is optionally substituted, or R 2 and R 3 together with the atom to which they are attached form a cycloalkyl or heterocyclyl; R 4 and R 5 are independently H, -(SO2)R 2 , -(SO2)NR 2 R 3 , -(CO)R 2 , -(CONR 2 R 3), aryl, arylheteroaryl, alkylenearyl, heteroaryl, cycloalkyl, heterocyclyl, alkyl, haloalkyl, and alkoxy, each of which is optionally substituted; or R 4 and R 5 together with the atom to which they are attached form a cycloalkyl or heterocyclyl, each of which is optionally substituted; R 9 is selected from the group consisting of H, C1-C6 alkyl, haloalkyl, cycloalkyl, and heterocyclyl; X 1 are S, O, NH, and NR 6 R 6 is selected from the group consisting of C1-C6 alkyl, alkoxy, haloalkyl, cycloalkyl, and heterocyclyl; Y is CR 2 , O, N, S, SO, and SO2, and when Y is O, S, SO, or SO2, R 5 does not exist, and R 4 and R 5 When taken together with the atom to which they are attached form a cycloalkyl or heterocyclyl, Y is CR 2 or N, n is selected from 0, 1, and 2.

[0155] In some embodiments of Formula (I), n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 0 or 1. In some embodiments, n is 1 or 2. In some embodiments, n is 0 or 2.

[0156] In some embodiments of formula (I), X 1 is O. In some embodiments, X 1 is S. In some embodiments, X 1 is NH. In some embodiments, X 1 is NR 6In some embodiments, X 1 S, O, and NR 6 In some embodiments, X is selected from the group consisting of 1 is selected from the group consisting of S, O, and NCH3. 1 is S or O, and in some embodiments, X 1 is S or NR 6 In some embodiments, R 6 is C1-C6 alkyl.

[0157] In some embodiments of Formula (I), R 2 and R 3 is H.

[0158] In some embodiments of formula (I), Y is N, CR 2 In some embodiments, Y is N or O. In some embodiments, Y is N. In some embodiments, Y is CR 2 In some embodiments, Y is O.

[0159] In some embodiments, R 4 and R 5 are independently H, -(SO2)R 2 , -(SO2)NR 2 R 3 , -(CO)R 2 , -(CONR 2 R 3 ), aryl, arylheteroaryl, heteroaryl, alkylenearyl, cycloalkyl, alkylenecycloalkyl, heterocyclyl, alkyleneheterocyclyl, alkyl, haloalkyl, and alkoxy, each of which is optionally substituted; or R 4 and R 5 together with the atom to which they are attached form a cycloalkyl or heterocyclyl, each of which is optionally substituted.

[0160] In some embodiments of Formula (I), R 4 is -C(O)-alkyl, -C(O)-cycloalkyl, -C(O)-aryl, -C(O)-heteroaryl, -(SO2)NR 2 R 3 , -SO2-alkyl, and -SO2-cycloalkyl, each of which is optionally substituted. In some embodiments, R 4 is -C(O)-alkyl, -C(O)-cycloalkyl, -SO2-alkyl, -SO2-haloalkyl, -SO2-cycloalkyl, and -(SO2)NR 2 R 3 In some embodiments of formula (I), R is selected from the group consisting of: 4 is selected from the group consisting of -SO2 alkyl, -SO2 haloalkyl, or -SO2 cycloalkyl. In some embodiments of Formula (I), R 4 is selected from the group consisting of -SOMe, -SOEt, and -SO-cPr. 2 and R 3 are each independently -C 1-5 In some embodiments, R 2 and R 3 together with the nitrogen atom to which they are attached form an optionally substituted heterocyclyl. In some embodiments, the optionally substituted heterocyclyl is morpholine, thiomorpholine, or thiomorpholine 1,1-dioxide.

[0161] In some embodiments of Formula (I), R 5 is aryl, heteroaryl, or cycloalkyl, each of which is optionally substituted.

[0162] In some embodiments, R 5 is aryl. In some embodiments, aryl is [ka] and R b is one or more selected from the group consisting of halogen, haloalkyl, alkyl, Oalkyl, Ohaloalkyl, alkylene-Ohaloalkyl, cycloalkyl, heterocyclylaryl, heteroaryl, alkylnitrile, or CN. In some embodiments, haloalkyl is selected from CF, CFCH, CHF, or CHF. In some embodiments, alkyl is selected from -C 1-5 In some embodiments, -C 1-5 Alkyl is methyl, ethyl, propyl, i-propyl, butyl, or t-butyl. In some embodiments, methyl, ethyl, propyl, i-propyl, butyl, or t-butyl is optionally substituted with OH. In some embodiments, cycloalkyl is C 3-6 In some embodiments, the aryl is phenyl. In some embodiments, the heteroaryl is a 5- or 6-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S. In some embodiments, the heterocyclyl is a 4- to 7-membered heterocyclyl having 1 or 2 heteroatoms selected from N, O, and S. In some embodiments, the O-haloalkyl is selected from OCF3, OCHF2, or OCH2F. In some embodiments, the O-alkyl is O-methyl, O-ethyl, O-propyl, Oi-propyl, O-butyl, or Ot-butyl.

[0163] In some embodiments, R 5is heteroaryl. In some embodiments, the heteroaryl is an optionally substituted 5-14 membered heteroaryl. In some embodiments, the heteroaryl is an optionally substituted 5-14 membered heteroaryl having 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the optionally substituted 5-14 membered heteroaryl is selected from the group consisting of pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, indolizinyl, azaindolyl, indolyl, azaindolyl, benzoxazolyl, benzthiazolyl, benzfuranyl, benzthiophenyl, imidazopyridinyl, imidazopyrazinyl, and benzimidazolyl. In some embodiments, the optionally substituted 5-14 membered heteroaryl is selected from the group consisting of pyrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzoxazolyl, imidazopyridinyl, and imidazopyrazinyl. 5 teeth, [ka] where R b is one or more selected from the group consisting of halogen, haloalkyl, alkyl, Oalkyl, Ohaloalkyl, alkylene-Ohaloalkyl, cycloalkyl, heterocyclylaryl, heteroaryl, alkylnitrile, or CN. In some embodiments, haloalkyl is selected from CF, CFCH, CHF, or CHF. In some embodiments, alkyl is selected from -C 1-5 In some embodiments, -C 1-5 Alkyl is methyl, ethyl, propyl, i-propyl, butyl, or t-butyl. In some embodiments, methyl, ethyl, propyl, i-propyl, butyl, or t-butyl is optionally substituted with OH. In some embodiments, cycloalkyl is C 3-6In some embodiments, the aryl is phenyl. In some embodiments, the heteroaryl is a 5- or 6-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S. In some embodiments, the heterocyclyl is a 4- to 7-membered heterocyclyl having 1 or 2 heteroatoms selected from N, O, and S. In some embodiments, the O-haloalkyl is selected from OCF3, OCHF2, or OCH2F. In some embodiments, the O-alkyl is O-methyl, O-ethyl, O-propyl, Oi-propyl, O-butyl, or Ot-butyl.

[0164] In some embodiments, R 5 is cycloalkyl. In some embodiments, cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each of which is optionally substituted. In some embodiments, optionally substituted cycloalkyl is [ka] It is.

[0165] In some embodiments, R 5 is selected from the group consisting of phenyl, 3-chlorophenyl, 3-chloro-4-fluorophenyl, 3-trifluoromethylphenyl, 3,4-difluorophenyl, and 2,6-difluorophenyl. 5 is cyclopropyl. In some embodiments, R 5 is selected from the group consisting of pyridin-3-yl and 1-methylindazol-6-yl. 5 is selected from the group consisting of H, phenyl, 3-chlorophenyl, 3-chloro-4-fluorophenyl, 3-trifluoromethylphenyl, 3,4-difluorophenyl, cyclopropyl, pyridin-3-yl, 1-methylindazol-6-yl, 3,3-difluorocyclobutyl, and 4,4-difluorocyclohexyl.5 is 3-chlorophenyl. In some embodiments, R 5 is H. In some embodiments, R 5 teeth, [ka] In some embodiments, R 5 is -CHCHPh. In some embodiments, R 5 is selected from the group consisting of H, aryl, heteroaryl, alkylenearyl, cycloalkyl, heterocyclyl, alkyl, and haloalkyl, each of which is optionally substituted; or R 4 and R 5 together with the atom to which they are attached form an optionally substituted heterocyclyl.

[0166] In some embodiments of Formula (I), R 5 is optionally substituted with one or more halogen, haloalkyl, alkyl, Oalkyl, Ohaloalkyl, cycloalkyl, heterocyclylaryl, or heteroaryl. In some embodiments, haloalkyl is selected from CF, CHF, or CHF. In some embodiments, alkyl is selected from -C 1-5 In some embodiments, -C 1-5 Alkyl is methyl, ethyl, propyl, i-propyl, butyl, or t-butyl. In some embodiments, cycloalkyl is 3-6In some embodiments, the aryl is phenyl. In some embodiments, the heteroaryl is a 5- or 6-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S. In some embodiments, the heterocyclyl is a 4- to 7-membered heterocyclyl having 1 or 2 heteroatoms selected from N, O, and S. In some embodiments, the O-haloalkyl is selected from OCF3, OCHF2, or OCH2F. In some embodiments, the O-alkyl is O-methyl, O-ethyl, O-propyl, Oi-propyl, O-butyl, or Ot-butyl.

[0167] In some embodiments of Formula (I), R 4 is H or -C 1-5 is alkyl, R 5 is aryl. In some embodiments, R 4 is H or -C 1-5 is alkyl, R 5 is heteroaryl. In some embodiments, R 4 is H or -C 1-5 is alkyl, R 5 is cycloalkyl. In some embodiments, -C 1-5 Alkyl is methyl, ethyl, or propyl. In some embodiments, -C 1-5Alkyl is methyl. In some embodiments, aryl is an optionally substituted phenyl. In some embodiments, heteroaryl is a 5-14 membered heteroaryl having 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the optionally substituted 5-14 membered heteroaryl is selected from the group consisting of pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, indolizinyl, azaindolyl, indolyl, azaindolyl, benzoxazolyl, benzthiazolyl, benzfuranyl, benzthiophenyl, imidazopyridinyl, imidazopyrazinyl, and benzimidazolyl. In some embodiments, heteroaryl is a 5 or 6 membered heteroaryl ring. In some embodiments, the 5-membered heteroaryl is an optionally substituted pyrazolyl, imidazolyl, or oxazolyl. In some embodiments, the 6-membered heteroaryl is an optionally substituted pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl. In some embodiments, the cycloalkyl is an optionally substituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments, the aryl is an optionally substituted halogen, C 1-6 Haloalkyl, C 1-6 Alkyl, OC 1-6 Alkyl, OC 1-6 In some embodiments, the heteroaryl is optionally substituted with one or more substituents selected from the group consisting of halogen, C 1-6 Haloalkyl, C 1-6 Alkyl, OC 1-6 Alkyl, OC 1-6 Optionally substituted with one or more substituents selected from the group consisting of haloalkyl, or C3-6 cycloalkyl.

[0168] In some embodiments of Formula (I), R 4 is -(CO)R 2and R 5 is aryl. In some embodiments, R 4 is -(CO)R 2 and R 5 is heteroaryl. In some embodiments, R 4 is -(CO)R 2 and R 5 is cycloalkyl. In some embodiments, the aryl is an optionally substituted phenyl. In some embodiments, the aryl is an optionally substituted phenyl. In some embodiments, the heteroaryl is a 5-14 membered heteroaryl having 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the optionally substituted 5-14 membered heteroaryl is selected from the group consisting of pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, indolizinyl, azaindolizinyl, indolyl, azaindolyl, benzoxazolyl, benzthiazolyl, benzfuranyl, benzthiophenyl, imidazopyridinyl, imidazopyrazinyl, and benzimidazolyl. In some embodiments, the heteroaryl is a 5-6 membered heteroaryl ring. In some embodiments, the 5-membered heteroaryl is an optionally substituted pyrazolyl, imidazolyl, oxazolyl, and in some embodiments, the 6-membered heteroaryl is an optionally substituted pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl. In some embodiments, the cycloalkyl is an optionally substituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments, the aryl is an optionally substituted halogen, C 1-6 Haloalkyl, C 1-6 Alkyl, OC 1-6 Alkyl, OC 1-6 In some embodiments, the heteroaryl is optionally substituted with one or more substituents selected from the group consisting of halogen, C 1-6Haloalkyl, C 1-6 Alkyl, OC 1-6 Alkyl, OC 1-6 Optionally substituted with one or more substituents selected from the group consisting of haloalkyl, or C3-6 cycloalkyl.

[0169] In some embodiments of Formula (I), R 4 is -(SO2)R 2 and R 5 is aryl. In some embodiments, R 4 is -(SO2)R 2 and R 5 is heteroaryl. In some embodiments, R 4 is -(SO2)R 2 and R 5is cycloalkyl. In some embodiments, aryl is an optionally substituted phenyl. In some embodiments, heteroaryl is a 5-14 membered heteroaryl having 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the optionally substituted 5-14 membered heteroaryl is selected from the group consisting of pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, quinolinyl, isoquinolinyl, quinoxalinyl, cinnolinyl, indolizinyl, azaindolizinyl, indolyl, azaindolyl, benzoxazolyl, benzthiazolyl, benzfuranyl, benzthiophenyl, imidazopyridinyl, imidazopyrazinyl, and benzimidazolyl. In some embodiments, heteroaryl is a 5-6 membered heteroaryl ring. In some embodiments, the 5-membered heteroaryl is an optionally substituted pyrazolyl, imidazolyl, or oxazolyl. In some embodiments, the 6-membered heteroaryl is an optionally substituted pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl. In some embodiments, the cycloalkyl is an optionally substituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments, the aryl is an optionally substituted halogen, C 1-6 Haloalkyl, C 1-6 Alkyl, OC 1-6 Alkyl, OC 1-6 In some embodiments, the heteroaryl is optionally substituted with one or more substituents selected from the group consisting of halogen, C 1-6 Haloalkyl, C 1-6 Alkyl, OC 1-6 Alkyl, OC 1-6 and optionally substituted with one or more substituents selected from the group consisting of haloalkyl, C3-6 cycloalkyl, and C3-6 cycloalkyl. 1-6 Haloalkyl is CF, CHF, or CHF. In some embodiments, OC 1-6Haloalkyl is OCF, OCHF, or OCHF. In some embodiments, cycloalkyl is halogen, C 1-6 Alkyl or OC 1-6 It is optionally substituted with alkyl.

[0170] In some embodiments of Formula (I), R 4 and R 5 together with the atom to which they are attached form a cycloalkyl or heterocyclyl. In some embodiments, R 4 and R 5 taken together with the atom to which they are attached form a cycloalkyl or heterocyclyl, each of which is optionally substituted. In some embodiments, the cycloalkyl or heterocyclyl is optionally substituted with -NS(O2)(alkyl)(aryl). In some embodiments, the alkyl is C 1-5 In some embodiments, the heterocyclyl is a 4- to 10-membered heterocyclyl. In some embodiments, the heterocyclyl is a saturated 4- to 7-membered heterocyclyl.

[0171] In some embodiments of Formula (I), n is 0 and R 4 and R 5 together with the atoms to which they are attached, [ka] In some embodiments, the optionally substituted heterocyclyl is selected from the group consisting of: [ka] In some embodiments, the optionally substituted heterocyclyl is [ka] In some embodiments, the optionally substituted heterocyclyl is [ka] It is.

[0172] In some embodiments of Formula (I), R 1 teeth, [ka] is selected from the group consisting of:

[0173] In some embodiments of Formula (I), R 1 teeth, [ka] In some embodiments, R 1 teeth, [ka] In some embodiments, R 1 teeth, [ka] In some embodiments, R 1 [ka]

[0174] In some embodiments of Formula (I), R a H, halo, C 1-3 In some embodiments, R a is H. In some embodiments, R a is C 1-3 In some embodiments, R a is haloalkyl. In some embodiments, halo is F. In some embodiments, C 1-3Alkylalkyl is methyl, ethyl, or isopropyl. In some embodiments, haloalkyl is CF3, CHF2, or CH2F.

[0175] In some embodiments of Formula (I), Y is CH and R 4 and R 5 is H.

[0176] In some embodiments of Formula (I), Y is N and R 4 is H and R 5 is ethyl optionally substituted with -N(S(O2)alkyl)(aryl) or -N(S(O2)cycloalkyl)(aryl). In some embodiments, alkyl is 1-5 alkyl, and cycloalkyl is C 3-6 cycloalkyl and aryl is phenyl optionally substituted with one or more halogen atoms.

[0177] In some embodiments of Formula (I), n is 1 and X 1 is O or N, Y is N, and R 1 teeth, [ka] and R 2 and R 3 is H and R 4 -H, -C 1-5 Alkyl, -C(O)alkyl, -C(O)cycloalkyl, -(SO2)NR 2 R 3 , -SO2 alkyl, -SO2 haloalkyl, and -SO2 cycloalkyl, each of which is optionally substituted; R 5 is aryl, heteroaryl, or cycloalkyl, each of which is optionally substituted.

[0178] In some embodiments of Formula (I), n is 1 and X 1 is O or N, Y is O, and R 1 teeth, [ka] and R 2 and R 3 is H and R 5 is aryl, heteroaryl, cycloalkyl, or alkylenecycloalkyl, each of which is optionally substituted.

[0179] In some embodiments of Formula (I), n is 0 and X 1 is O or N, Y is N, and R 1 teeth, [ka] and R 4 and R 5 together with the atom to which they are attached form a cycloalkyl or heterocyclyl, each of which is optionally substituted.

[0180] In some embodiments, the disclosure provides a compound of formula (Ia) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony,

[0181] R 1 , R 2 , R 3 , R 4 , R 5 , R a , X 1 , n, and Y are as defined above for formula (I).

[0182] In some embodiments of Formula (Ia), R 1 teeth, [ka] where n is 1, Y is N, and X 1 is S or O, and the variable R2 , R 3 , R 4 , R 5 , and R a is as defined above for formula (I).

[0183] In some embodiments of Formula (Ia), n is 1 and X 1 is S, Y is N, and R 1 teeth, [ka] and R 2 and R 3 is H and R 4 is -SO2 alkyl, -SO2 haloalkyl, or -SO2 cycloalkyl, each of which is optionally substituted; R 5 is heteroaryl, each of which is optionally substituted; R a is H or F. In some further embodiments, R 4 -SO2C 1-5 alkyl, -SO2cyclopropyl, -SO2CF3 or -SO2CHF2, and the heteroaryl is an optionally substituted pyridine or pyrazine. In some embodiments, the heteroaryl is an optionally substituted pyridine.

[0184] In some embodiments of Formula (Ia), n is 1 and X 1 is S, Y is N, and R 1 teeth, [ka] and R 2 and R 3 is H and R 4 is -SOMe, -SOEt, or -SOcyclopropyl, each of which is optionally substituted; R 5 is pyridine or pyrazine, each of which is optionally substituted; R ais H. In some embodiments, R 5 is an optionally substituted pyridine.

[0185] In some embodiments of Formula (Ia), n is 1 and X 1 is S, Y is N, and R 1 teeth, [ka] and R 2 and R 3 is H and R 4 is -SO2 alkyl or -SO2 cycloalkyl, each of which is optionally substituted; R 5 teeth, [ka] and In the formula, R b is halogen, -C 1-5 Alkyl, haloalkyl, -OC 1-5 is selected from the group consisting of alkyl, -OHaloalkyl, -CHOHaloalkyl, cyclopropyl, and CN; a is H. In some embodiments, halogen is F or Cl. In some embodiments, haloalkyl is CF, CHF, CHCF, or CFCH. In some embodiments, -C 1-5 Alkyl is methyl.

[0186] In some embodiments of Formula (Ia), n is 1 and X 1 is S, Y is N, and R 1 is [ka] R 2 and R 3 is H and R 4 is -SOMe, -SOEt, or -SOcyclopropyl, each of which is optionally substituted; R5 teeth, [ka] and In the formula, R b is halogen, -C 1-5 Alkyl, haloalkyl, -OC 1-5 alkyl, -OHaloalkyl, -CHOHaloalkyl, cyclopropyl, or CN; R a is H. In some embodiments, halogen is F or Cl. In some embodiments, haloalkyl is CF, CHF, CHCF, or CFCH. In some embodiments, -C 1-5 Alkyl is methyl.

[0187] In some embodiments of Formula (Ia), n is 1 and X 1 is S, Y is N, and R 1 is [ka] R 2 and R 3 is H and R 4 is -SOMe, -SOEt, or -SOcyclopropyl, each of which is optionally substituted; R 5 is [ka] In the formula, R b is selected from the group consisting of Cl, F, Me, cyclopropyl, CF, CHF, CFCH, OCF, OCHF, OCHCFH, and CN; R a is H.

[0188] In some embodiments, the disclosure provides a compound of formula (Ib) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony, R 1 , R 2 , R 3 , R 4 , R 5 , R a , X 1 , n, and Y are as defined above for formula (I).

[0189] In some embodiments of Formula (I)-(Ib), each optionally substituted alkyl is independently an optionally substituted C 1-6 In some embodiments, C 1-6 Alkyl is Me or Et.

[0190] In some embodiments of Formula (I)-(Ib), each optionally substituted haloalkyl independently represents an optionally substituted C 1-6 In some embodiments, C is haloalkyl. 1-6 Haloalkyl is CF, CHF, or CHF. In some embodiments, C 1-6 Haloalkyl is CF3 or CHF2.

[0191] In some embodiments of Formula (I)-(Ib), each optionally substituted cycloalkyl can independently be selected from the group consisting of optionally substituted C 3-12 In some embodiments, cycloalkyl is C 3-6 In some embodiments, cycloalkyl is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0192] In some embodiments of Formula (I)-(Ib), each optionally substituted heterocyclyl is independently an optionally substituted 3-12 membered heterocycloalkyl having 1 or 2 heteroatoms independently selected from N, O, and S. In some embodiments, each optionally substituted heterocyclyl is independently an optionally substituted 3-6 membered heterocycloalkyl having 1 or 2 heteroatoms independently selected from N, O, and S. In further embodiments, the heterocycloalkyl is an optionally substituted 5- or 6-membered heterocycle having 1 or 2 heteroatoms independently selected from N, O, and S. In some embodiments, the heterocyclyl is selected from the group consisting of aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, and morpholinyl, and thiomorpholinyl.

[0193] In some embodiments of Formula (I)-(Ib), each optionally substituted aryl is independently selected from the group consisting of: 6-12 In a further embodiment, C 6-12 Aryl is optionally substituted phenyl.

[0194] In some embodiments of Formula (I)-(Ib), each optionally substituted heteroaryl is independently a 5-12 membered heteroaryl having 1, 2, or 3 heteroatoms independently selected from N, O, and S. In some embodiments, each optionally substituted heteroaryl is independently a 5-12 membered heteroaryl having 3 heteroatoms independently selected from N, O, and S. In some embodiments, each optionally substituted heteroaryl is independently a 5-12 membered heteroaryl having 2 heteroatoms independently selected from N, O, and S. In some embodiments, each optionally substituted heteroaryl is independently a 5-12 membered heteroaryl having 1 heteroatom independently selected from N, O, and S. In further embodiments, each optionally substituted heteroaryl is an optionally substituted 5- or 6-membered heteroaryl having 1 heteroatom independently selected from N, O, and S. In some embodiments, each heteroaryl is independently selected from the group consisting of tetrazole, oxadiazole, thiadiazole, imidazole, pyrazole, thiazole, or oxazole, each of which is optionally substituted.

[0195] In some embodiments, the compound of formula (I) is [Table 21-1] [Table 21-2] [Table 21-3] [Table 21-4] [Table 21-5] [Table 21-6]

Table 21-7

Table 21-8

Table 21-9

Table 21-10

Table 21-11

Table 21-12

Table 21-13

Table 21-14

Table 21-15

Table 21-16

Table 21-17

Table 21-18

Table 21-19

Table 21-20

Table 21-21

Table 21-22

Table 21-23

[0196] In some embodiments, the present disclosure provides a compound of formula (Ic) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony, R a is H, Me, or F; R 4 and R 5 is as defined above in formula (I).

[0197] In some embodiments of Formula (Ic), R a is H. In some embodiments, R a is F. In some embodiments, R a is Me.

[0198] In some embodiments of Formula (Ic), R 4is selected from the group consisting of alkylenealkoxy, alkyleneheterocyclyl, -S(O)alkyl, -S(O)cycloalkyl, -S(O)alkylenecycloalkyl, -S(O)alkyleneheterocyclyl, -S(O)N(H)alkyleneheterocyclyl, -C(O)alkyl, -C(O)cycloalkyl, -C(O)alkylenecycloalkyl, -C(O)alkyleneheterocyclyl, and -C(O)N(H)alkyleneheterocyclyl. In some embodiments, R 4 is selected from the group consisting of alkyleneheterocyclyl, -S(O)2 alkyl, -S(O)2 cycloalkyl, -S(O)2 alkyleneheterocyclyl, -C(O)alkyleneheterocyclyl, and -C(O)N(H)alkyleneheterocyclyl. 4 is selected from the group consisting of -S(O)2 alkyl, -S(O)2 cycloalkyl, and -S(O)2 alkyleneheterocyclyl. 4 is -S(O)alkyl. In some embodiments, R 4 is -S(O)cycloalkyl. In some embodiments, R 4 is -S(O)N(H)alkyleneheterocyclyl. In some embodiments, alkylene is C 1-5 In some embodiments, the alkylene is an optionally substituted 4-10 membered heterocyclyl having 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the alkylene is 1-5 In some embodiments, the alkylene is an optionally substituted 4-7 membered heterocyclyl having 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the alkylene is 2-4and the heterocyclyl is an optionally substituted 6-membered heterocyclyl having 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the heterocyclyl is selected from the group consisting of piperidine, morpholine, thiomorpholine, thiomorpholine 1-oxide, thiomorpholine 1,1-dioxide, and piperizine, each of which is optionally substituted. In some embodiments, the optional substituents are selected from the group consisting of alkyl, haloalkyl, alkoxy, acyl, sulfonyl, heteroaryl, and heterocyclyl.

[0199] In some embodiments of Formula (Ic), R 5 teeth, [ka] In some embodiments, R 5 teeth, [ka] In some embodiments, R 5 teeth, [ka] In some embodiments, R 5 teeth, [ka] In some embodiments, R 5 teeth, [ka] In some embodiments, R b is selected from the group consisting of halogen, haloalkyl, alkyl, Oalkyl, Ohaloalkyl, alkylene-Ohaloalkyl, cycloalkyl, heterocyclylaryl, heteroaryl, alkylnitrile, or CN. bis selected from the group consisting of halo, alkyl, haloalkyl, alkoxy, haloalkoxy, acyl, sulfonyl, cycloalkyl, heteroaryl, and heterocyclyl. In some embodiments, haloalkyl is selected from CF, CFCH, CHF, or CHF. In some embodiments, alkyl is selected from -C 1-5 In some embodiments, -C 1-5 Alkyl is methyl, ethyl, propyl, i-propyl, butyl, or t-butyl. In some embodiments, methyl, ethyl, propyl, i-propyl, butyl, or t-butyl is optionally substituted with OH. In some embodiments, cycloalkyl is C 3-6 In some embodiments, the aryl is phenyl. In some embodiments, the heteroaryl is a 5- or 6-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S. In some embodiments, the heterocyclyl is a 4- to 7-membered heterocyclyl having 1 or 2 heteroatoms selected from N, O, and S. In some embodiments, the O-haloalkyl is selected from OCF3, OCHF2, or OCH2F. In some embodiments, the O-alkyl is O-methyl, O-ethyl, O-propyl, Oi-propyl, O-butyl, or Ot-butyl. In some embodiments, R b is selected from the group consisting of F, Cl, -CH3, -CH2CH3, -CF3, -CHF2, -CF2CH3, -CN, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCHF2, -OCH2CF2H, and cyclopropyl. In some embodiments, m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2.

[0200] In some embodiments, the disclosure provides a compound of formula (Id) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony, U is NR d , O, S, S(O), S(O)2, CH2, CHF, or CF2; R a is H, Me, or F; R b are each independently halo, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), -S(O2)R e , cycloalkyl, heteroaryl, or heterocyclyl; R c are each independently F, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), -S(O2)R e , heteroaryl, or heterocyclyl, and / or two R c The C groups, together with the carbon atoms to which they are attached, are bridged or fused 3-7 forming a cycloalkyl, a bridged or fused 4- to 7-membered heterocyclyl, or a 5- or 6-membered heteroaryl, each of which is optionally substituted; R d is H, alkyl, acyl, sulfonyl, cycloalkyl, aryl, or heteroaryl; R e and R e’ are each independently H, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CHcycloalkyl, -CHheterocyclyl, -CHaryl, or -CHheteroaryl; m is 0, 1, 2, or 3; p is 0, 1, 2, or 3; q is 0, 1, or 2; r is 1, 2, 3, or 4.

[0201] In some embodiments, the present disclosure provides a compound of formula (Ie) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony, U is NR d , O, S, S(O), S(O)2, CH2, CHF, or CF2; R a is H, Me, or F; R b are each independently halo, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), sulfonyl, cycloalkyl, heteroaryl, or heterocyclyl; R c are each independently F, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), -S(O2)R e , heteroaryl, or heterocyclyl, and / or two R c The C groups, together with the carbon atoms to which they are attached, are bridged or fused 3-7 forming a cycloalkyl, a bridged or fused 4- to 6-membered heterocyclyl, or a 5- or 6-membered heteroaryl, each of which is optionally substituted; R d is H, alkyl, acyl, sulfonyl, cycloalkyl, aryl, or heteroaryl; R e and R e’ are each independently H, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CHcycloalkyl, -CHheterocyclyl, -CHaryl, or -CHheteroaryl; m is 0, 1, 2, or 3; p is 0, 1, 2, or 3; q is 0, 1, or 2; r is 1, 2, 3, or 4.

[0202] In some embodiments, the disclosure provides a compound of formula (If) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony, U is NR d , O, S, S(O), S(O)2, CH2, CHF, or CF2; R a is H, Me, or F; R b are each independently halo, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), sulfonyl, cycloalkyl, heteroaryl, or heterocyclyl; R c are each independently F, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), -S(O2)R e , heteroaryl, or heterocyclyl, and / or two R c The C groups, together with the carbon atoms to which they are attached, are bridged or fused 3-7 forming a cycloalkyl, a bridged or fused 4- to 7-membered heterocyclyl, or a 5- or 6-membered heteroaryl, each of which is optionally substituted; R d is H, alkyl, acyl, sulfonyl, cycloalkyl, aryl, or heteroaryl; R e and R e’are each independently H, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CHcycloalkyl, -CHheterocyclyl, -CHaryl, or -CHheteroaryl; m is 0, 1, 2, or 3; p is 0, 1, 2, or 3; q is 0, 1, or 2; r is 1, 2, 3, or 4.

[0203] In some embodiments, the disclosure provides a compound of formula (Ig) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony, U is NR d , O, S, S(O), S(O)2, CH2, CHF, or CF2; R a is H, Me, or F; R b are each independently halo, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), sulfonyl, cycloalkyl, heteroaryl, or heterocyclyl; R c are each independently F, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), -S(O2)R e , heteroaryl, or heterocyclyl, and / or two R c The C groups, together with the carbon atoms to which they are attached, are bridged or fused 3-7 forming a cycloalkyl, a bridged or fused 4- to 7-membered heterocyclyl, or a 5- or 6-membered heteroaryl, each of which is optionally substituted; Rd is H, alkyl, acyl, sulfonyl, cycloalkyl, aryl, or heteroaryl; R e and R e’ are each independently H, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CHcycloalkyl, -CHheterocyclyl, -CHaryl, or -CHheteroaryl; m is 0, 1, 2, or 3; p is 0, 1, 2, or 3; q is 0, 1, or 2; r is 1, 2, 3, or 4.

[0204] In some embodiments, the compound has the formula: [ka] wherein U, R a , R b , m, and r are as defined above in formulas (Id), (Ie), (If), and (Ig); V is O or NR d It is.

[0205] In some embodiments of Formulas (Id)-(Ig) and (Id-1)-(Ig-1), U is NR d , O, or S and V is O. In some embodiments, U is N, O, or S and V is NR d In some embodiments, U is NR d and V is NR d In some embodiments, U is O and V is NR d In some embodiments, U is S and V is NR d In some embodiments, U is NR d and V is O. In some embodiments, U is O and V is O. In some embodiments, U is S and V is O.

[0206] In some embodiments of Formulas (Id)-(Ig) and (Id-1)-(Ig-1), U is O, S, S(O), CH, or NR d In some embodiments, U is O, S, CH, or NR d In some embodiments, U is O, S, or NR d In some embodiments, U is O or CH. In some embodiments, U is O. In some embodiments, U is S. In some embodiments, U is NR d In some embodiments, U is S(O).

[0207] In some embodiments of Formulas (Id)-(Ig) and (Id-1)-(Ig-1), R a is H. In some embodiments, R a is F. In some embodiments, R a is Me.

[0208] In some embodiments of Formulas (Id)-(Ig) and (Id-1)-(Ig-1), R b is halo, alkyl, haloalkyl, alkyl, haloalkoxy, cycloalkyl, heterocyclyl, heteroaryl, or nitrile. b is halo, alkyl, haloalkyl, alkyl, haloalkoxy, cycloalkyl, or nitrile. In some embodiments, haloalkyl is selected from CF, CFCH, CHF, or CHF. In some embodiments, alkyl is -C 1-5 In some embodiments, -C 1-5 Alkyl is methyl, ethyl, propyl, i-propyl, butyl, or t-butyl. In some embodiments, cycloalkyl is 3-6In some embodiments, the heteroaryl is a 5- or 6-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S. In some embodiments, the heterocyclyl is a 4- to 7-membered heterocyclyl having 1 or 2 heteroatoms selected from N, O, and S. In some embodiments, the haloalkoxy is selected from OCF3, OCHF2, or OCH2F. In some embodiments, the alkoxy is O-methyl, O-ethyl, O-propyl, Oi-propyl, O-butyl, or Ot-butyl. In some embodiments, R b is -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ).

[0209] In some embodiments of Formulas (Id)-(Ig), R c , F, C 1-5 Alkyl, haloalkyl, C 1-5 Alkoxy, haloalkoxy, acyl, sulfonyl, 5- or 6-membered heteroaryl, or C 3-6 In some embodiments, R c is -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ In some embodiments, two R c The C groups, together with the carbon atoms to which they are attached, are bridged or fused 3-7 In some embodiments, two R c Bridged or fused C groups, which together with the carbon atoms to which they are attached, are optionally substituted. 3-7 In some embodiments, two R cIn some embodiments, two R c The groups, together with the carbon atoms to which they are attached, form an alkoxy or aminoalkyl bridge. In some embodiments, the optional substituents are one or more R b In some embodiments, the optional substituents are F, C 1-5 Alkyl, C 1-5 Alkoxy, CF3, CF2H, CFH2, -OCF3, -OCF2H, -OCFH2, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), and -SO2R e In some embodiments, the optional substituents are selected from the group consisting of F, C 1-5 Alkyl, C 1-5 In some embodiments, the optional substituents are selected from the group consisting of F or C. 1-5 In some embodiments, the optional substituent is F. In some embodiments, the optional substituent is C 1-5 In some embodiments, C 1-5 Alkyl is methyl. In some embodiments, C 1-5 Alkyl is ethyl. In some embodiments, C 1-5 Alkyl is propyl. In some embodiments, C 1-5 Alkyl is isopropyl.

[0210] In some embodiments of Formulas (Id)-(Ig) and (Id-1)-(Ig-1), R e and R e’ are each independently H, alkyl, cycloalkyl, or -CHcycloalkyl. In some embodiments, alkyl is -C 1-5In some embodiments, -C 1-5 Alkyl is methyl, ethyl, propyl, i-propyl, butyl, or t-butyl. In some embodiments, cycloalkyl is 3-6 In some embodiments, cycloalkyl is cyclopropyl. In some embodiments, R e and R e’ is H.

[0211] In some embodiments of Formulas (Id)-(Ig) and (Id-1)-(Ig-1), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2.

[0212] In some embodiments of Formula (Id)-(Ig), p is 0, 1, or 2. In some embodiments, p is 0 or 1. In some embodiments, p is 1 or 2. In some embodiments, p is 0. In some embodiments, p is 1. In some embodiments, p is 2.

[0213] In some embodiments of Formulas (Id)-(Ig) and (Id-1)-(Ig-1), r is 1, 2, or 3. In some embodiments, r is 1 or 2. In some embodiments, r is 2 or 3. In some embodiments, r is 1. In some embodiments, r is 2. In some embodiments, r is 3. In some embodiments, r is 4.

[0214] In some embodiments of Formula (Id)-(Ig), q is 0 or 1. In some embodiments, q is 0. In some embodiments, q is 1. In some embodiments, q is 2.

[0215] In some embodiments of Formula (Id)-(Ig), r is 1 and p is 1. In some embodiments, r is 2 and p is 1. In some embodiments, r is 3 and p is 1.

[0216] In some embodiments, the present disclosure provides a compound of formula (Ih) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony, U is NR d , O, S, S(O), S(O)2, CH2, CHF, or CF2; X 1 , X 2 , X 3 , and X 4 are each independently CH or N; R a is H, Me, or F; R b are each independently halo, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), -SO2R e , cycloalkyl, heteroaryl, or heterocyclyl; R c are each independently F, alkyl, haloalkyl, alkoxy, or haloalkoxy, and / or two R c The groups, together with the atoms to which they are attached, are optionally substituted C 3-7 Forming a cycloalkyl, R d is H, alkyl, acyl, sulfonyl, cycloalkyl, aryl, or heteroaryl; R e and R e’are each independently H, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CHcycloalkyl, -CHheterocyclyl, -CHaryl, or -CHheteroaryl; m is 0, 1, 2, or 3; p is 0, 1, 2, or 3; q is 0, 1, or 2.

[0217] In some embodiments, the present disclosure provides a compound of formula (Ii) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony, U is NR d , O, S, S(O), S(O)2, CH2, CHF, or CF2; X 1 , X 2 , X 3 , and X 4 are each independently CH or N; R a is H, Me, or F; R b are each independently halo, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), -SO2R e , cycloalkyl, heteroaryl, or heterocyclyl; R c are each independently F, alkyl, haloalkyl, alkoxy, or haloalkoxy, and / or two R c The groups, together with the atoms to which they are attached, are optionally substituted C 3-7 Forming a cycloalkyl, R d H, alkyl, -C(O)R e , sulfonyl, cycloalkyl, aryl, or heteroaryl; R e and Re’ are each independently H, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CHcycloalkyl, -CHheterocyclyl, -CHaryl, or -CHheteroaryl; m is 0, 1, 2, or 3; p is 0, 1, 2, or 3; q is 0, 1, or 2.

[0218] In some embodiments, the disclosure provides a compound of formula (Ij) or a pharma- ceutically acceptable salt thereof: [ka] During the ceremony, U is NR d , O, S, S(O), S(O)2, CH2, CHF, or CF2; X 1 , X 2 , X 3 , and X 4 are each independently CH or N; R a is H, Me, or F; R b are each independently halo, alkyl, haloalkyl, alkoxy, haloalkoxy, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), -SO2R e , cycloalkyl, heteroaryl, or heterocyclyl; R c are each independently F, alkyl, haloalkyl, alkoxy, or haloalkoxy, and / or two R c The groups, together with the atoms to which they are attached, are optionally substituted C 3-7 Forming a cycloalkyl, R d H, alkyl, -C(O)R e , sulfonyl, cycloalkyl, aryl, or heteroaryl; Re and R e’ are each independently H, alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -CHcycloalkyl, -CHheterocyclyl, -CHaryl, or -CHheteroaryl; m is 0, 1, 2, or 3; p is 0, 1, 2, or 3; q is 0, 1, or 2.

[0219] In some embodiments of formula (Ih)-(Ij), NR d , O, S, S(O)2, or CH2. In some embodiments, U is NR d , O, S, or CH2. In some embodiments, U is O or CH2. In some embodiments, U is O. In some embodiments, U is CH2. In some embodiments, U is S. In some embodiments, U is S(O)2. In some embodiments, U is NR d It is.

[0220] In some embodiments of formulas (Ih)-(Ij), X 1 , X 2 , X 3 , and X 4 Each of X is CH. In some embodiments, 1 , X 2 , X 3 , and X 4 In some embodiments, one of X is N. 1 , X 2 , X 3 , and X 4 Two of X are N. 1 is N and X 2 , X 3 , and X 4 Each of X is CH. In some embodiments, 2 is N and X 1 , X 3 , and X 4 Each of X is CH. In some embodiments, 3is N and X 1 , X 2 , and X 4 Each of X is CH. In some embodiments, 4 is N and X 1 , X 2 , and X 3 Each of is CH.

[0221] In some embodiments of Formula (Ih)-(Ij), U is CH and X 1 , X 2 , X 3 , and X 4 In some embodiments, U is CH and one of X is N. 1 is N and X 2 , X 3 , and X 4 Each of X is CH. In some embodiments, U is CH and X 2 is N and X 1 , X 3 , and X 4 Each of X is CH. In some embodiments, U is CH and X 3 is N and X 1 , X 2 , and X 4 Each of X is CH. In some embodiments, U is CH and X 4 is N and X 1 , X 2 , and X 3 Each of is CH. In some embodiments, p is 0. In some embodiments, p is 1.

[0222] In some embodiments of Formula (Ih)-(Ij), U is O and X 1 , X 2 , X 3 , and X 4 In some embodiments, U is O and one of X is N. 1 is N and X 2 , X 3 , and X 4Each of X is CH. In some embodiments, U is O and X 2 is N and X 1 , X 3 , and X 4 Each of X is CH. In some embodiments, U is O and X 3 is N and X 1 , X 2 , and X 4 Each of X is CH. In some embodiments, U is O and X 4 is N and X 1 , X 2 , and X 3 Each of is CH.

[0223] In some embodiments of Formulas (Ih)-(Ij), R a is H. In some embodiments, R a is F. In some embodiments, R a is Me.

[0224] In some embodiments of Formulas (Ih)-(Ij), R b is halo, alkyl, haloalkyl, alkyl, haloalkoxy, cycloalkyl, heterocyclyl, heteroaryl, or nitrile. b is halo, alkyl, haloalkyl, alkyl, haloalkoxy, cycloalkyl, or nitrile. In some embodiments, haloalkyl is selected from CF, CFCH, CHF, or CHF. In some embodiments, alkyl is -C 1-5 In some embodiments, -C 1-5 Alkyl is methyl, ethyl, propyl, i-propyl, butyl, or t-butyl. In some embodiments, cycloalkyl is 3-6In some embodiments, the heteroaryl is a 5- or 6-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S. In some embodiments, the heterocyclyl is a 4- to 7-membered heterocyclyl having 1 or 2 heteroatoms selected from N, O, and S. In some embodiments, the haloalkoxy is selected from OCF3, OCHF2, or OCH2F. In some embodiments, the alkoxy is O-methyl, O-ethyl, O-propyl, Oi-propyl, O-butyl, or Ot-butyl.

[0225] In some embodiments of Formulas (Ih)-(Ij), R c , F, C 1-5 Alkyl, haloalkyl, C 1-5 Alkoxy, haloalkoxy, acyl, sulfonyl, 5- or 6-membered heteroaryl, or C 3-6 In some embodiments, R c , F, C 1-5 Alkyl, haloalkyl, C 1-5 In some embodiments, R c is F or C 1-5 In some embodiments, R c is F or methyl. In some embodiments, R c is F. In some embodiments, R c is methyl. In some embodiments, two R c In some embodiments, two R c C groups, together with the atoms to which they are attached, are optionally substituted 3-6 In some embodiments, two R c In some embodiments, the optional substituents are one or more R , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 21 , R 22 , R 23 , R 24 , R 25 , R 26 , R 27 , R 28 , R 29 , R 30 , R 31 , R 32 , R 33 , R 34 , R 35 , R 36 , R 37 , R 38 , R 39 , R 40 , R 41 , R 42 , Rb In some embodiments, the optional substituents are F, C 1-5 Alkyl, C 1-5 Alkoxy, CF3, CF2H, CFH2, -OCF3, -OCF2H, -OCFH2, -C(O)R e , -C(O)OR e , -C(O)N(R e )(R e’ ), and -SO2R e In some embodiments, the optional substituents are selected from the group consisting of F, C 1-5 Alkyl, C 1-5 In some embodiments, the optional substituents are selected from the group consisting of F or C. 1-5 In some embodiments, the optional substituent is F. In some embodiments, the optional substituent is C 1-5 In some embodiments, C 1-5 Alkyl is methyl. In some embodiments, C 1-5 Alkyl is ethyl. In some embodiments, C 1-5 Alkyl is propyl. In some embodiments, C 1-5 Alkyl is isopropyl. In some embodiments, two optional substituents are attached to the same carbon atom, which may also be referred to as germinal substitution.

[0226] In some embodiments of Formulae (Ih)-(Ij), U is NR d When R d and R c together with the atoms to which they are attached form a 5- to 7-membered heterocyclyl. d and R c together with the atom to which they are attached form a 6-membered heterocyclyl. In some embodiments, the heterocyclyl contains 1 or 2 heteroatoms selected from N, O, and S.

[0227] In some embodiments, the disclosure provides a compound of formula (Ih-1), formula (Ii-1), or formula (Ij-1): [ka] In the formula, R a , R b , R c , X 1 , X 2 , X 3 , X 4 , U, and m are as defined above in Formula (Ih), Formula (Ii), and Formula (Ij).

[0228] In some embodiments of Formula (Ih-1), Formula (Ii-1), and Formula (Ij-1), each R c is F. In some embodiments, each R c is Me. In some embodiments, two R c C groups, together with the carbon atoms to which they are attached, are optionally substituted 3-6 In some embodiments, two R c In some embodiments, the two R c The groups taken together with the carbon atom to which they are attached form an optionally substituted cyclopropyl. In some embodiments, the optional substituents are F or C. 1-5 In some embodiments, the optional substituent is F. In some embodiments, the optional substituent is C 1-5 In some embodiments, C 1-5 Alkyl is methyl. In some embodiments, C 1-5 Alkyl is ethyl. In some embodiments, C 1-5 Alkyl is propyl. In some embodiments, C 1-5Alkyl is isopropyl. In some embodiments, two optional substituents are attached to the same carbon atom, which may also be referred to as germinal substitution.

[0229] In some embodiments, R d is H, alkyl, or cycloalkyl. In some embodiments, R d is H. In some embodiments, R d is alkyl. In some embodiments, R d is cycloalkyl. In some embodiments, alkyl is methyl, ethyl, propyl, isopropyl, or t-butyl. In some embodiments, cycloalkyl is cyclopropyl, cyclopentyl, or cyclohexyl.

[0230] In some embodiments, m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2.

[0231] In some embodiments, p is 0, 1, or 2. In some embodiments, p is 0 or 1. In some embodiments, p is 1 or 2. In some embodiments, p is 0. In some embodiments, p is 1. In some embodiments, p is 2.

[0232] In some embodiments, q is 0 or 1. In some embodiments, q is 0. In some embodiments, q is 1. In some embodiments, q is 2.

[0233] In some embodiments, the HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, X1 is S, R a H, halogens, and C 1-3 is selected from the group consisting of alkyl, R 1 teeth, [ka] and R 2 is selected from the group consisting of alkyl, alkoxy, and cycloalkyl, each of which is optionally substituted; R 3 is H or alkyl, R 4 is alkyl, -(SO2)R 2 , -(SO2)NR 2 R 3 , and -(CO)R 2 is selected from the group consisting of R 5 is aryl or heteroaryl, or R 4 and R 5 together with the atom to which they are attached form a heterocyclyl, each of which is optionally substituted.

[0234] In some embodiments, R a is H.

[0235] In some embodiments, R 1 is as follows: [ka]

[0236] In some embodiments, R 4 is -(SO2)R 2 It is.

[0237] In some embodiments, -(SO)R 2is -(SO2)alkyl, -(SO2)alkyleneheterocyclyl, -(SO2)haloalkyl, -(SO2)haloalkoxy, or -(SO2)cycloalkyl.

[0238] In some embodiments, R 5 is heteroaryl.

[0239] In some embodiments, the heteroaryl is a 5-6 membered heteroaryl.

[0240] In some embodiments, the 5-6 membered heteroaryl is [ka] wherein R b is halogen, alkyl, alkoxy, cycloalkyl, —CN, haloalkyl, or haloalkoxy; and m is 0 or 1.

[0241] In some embodiments, R b are F, Cl, -CH3, -CH2CH3, -CF3, -CHF2, -CF2CH3, -CN, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCF3, -OCHF2, -OCH2CF2H, and cyclopropyl.

[0242] In some embodiments, aryl is selected from the group consisting of phenyl, 3-chlorophenyl, 3-chloro-4-fluorophenyl, 3-trifluoromethylphenyl, 3,4-difluorophenyl, and 2,6-difluorophenyl.

[0243] In some embodiments, the HDAC6 inhibitor has the formula (Ik): [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R bis H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted.

[0244] In some embodiments, R b is H, halogen, haloalkyl, or haloalkoxy.

[0245] In some embodiments, R 4 is optionally substituted alkyl or cycloalkyl.

[0246] In some embodiments, the HDAC6 inhibitor has the structure: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted.

[0247] In some embodiments, R b is H, halogen, haloalkyl, or haloalkoxy.

[0248] In some embodiments, R 4 is optionally substituted alkyl or cycloalkyl.

[0249] In some embodiments, R 4 is alkyl.

[0250] In some embodiments, the HDAC6 inhibitor has the structure: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted.

[0251] In some embodiments, R b is H, halogen, haloalkyl, or haloalkoxy.

[0252] In some embodiments, R 4 is optionally substituted alkyl.

[0253] In some embodiments, the HDAC6 inhibitor has the formula: [ka] A compound having the formula or a pharma- ceutically acceptable salt thereof; During the ceremony, X 1 is S, R a H, halogens, and C 1-3 is selected from the group consisting of alkyl, R 1 teeth, [ka] and R 2 is selected from the group consisting of alkyl, alkoxy, and cycloalkyl, each of which is optionally substituted; R 3 is H or alkyl, R 4 is alkyl, -(SO2)R 2 , -(SO2)NR 2 R3 , and -(CO)R 2 is selected from the group consisting of R 5 is aryl or heteroaryl, or R 4 and R 5 together with the atom to which they are attached form a heterocyclyl, each of which is optionally substituted.

[0254] In some embodiments of formula I(y), R a is H.

[0255] In some embodiments of formula I(y), R 1 teeth, [ka] It is.

[0256] In some embodiments of formula I(y), R 4 is -(SO2)R 2 It is.

[0257] In some embodiments of Formula I(y), -(SO2)R 2 is -(SO2)alkyl, -(SO2)alkyleneheterocyclyl, -(SO2)haloalkyl, -(SO2)haloalkoxy, or -(SO2)cycloalkyl.

[0258] In some embodiments of formula I(y), R 5 is heteroaryl.

[0259] In some embodiments of Formula I(y), the heteroaryl is a 5-6 membered heteroaryl.

[0260] In some embodiments of Formula I(y), the 5-6 membered heteroaryl is [ka] wherein R bis halogen, alkyl, alkoxy, cycloalkyl, —CN, haloalkyl, or haloalkoxy; and m is 0 or 1.

[0261] In some embodiments of formula I(y), R b are F, Cl, -CH3, -CH2CH3, -CF3, -CHF2, -CF2CH3, -CN, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCF3, -OCHF2, -OCH2CF2H, and cyclopropyl.

[0262] In some embodiments of Formula I(y), aryl is selected from the group consisting of phenyl, 3-chlorophenyl, 3-chloro-4-fluorophenyl, 3-trifluoromethylphenyl, 3,4-difluorophenyl, and 2,6-difluorophenyl.

[0263] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0264] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0265] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0266] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0267] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0268] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0269] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0270] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0271] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0272] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0273] In some embodiments, the fluoroalkyl-oxadiazole derivative is TYA-018 or an analog thereof. The structure of TYA-018 is: [ka]

[0274] Analogs of TYA-018 include, but are not limited to, the compounds listed in Table 2. [Table 2-1] [Table 2-2]

[0275] 5-Fluoronicotinamide Derivatives In some embodiments, the HDAC6 inhibitor is a 5-fluoronicotinamide derivative. Exemplary 5-fluoronicotinamide derivatives that can be used as HDAC6 inhibitors include those described herein and those disclosed in International Patent Application Publication No. PCT / US2020 / 054134, published as WO2021 / 067859(A1), the contents of which are incorporated herein by reference in their entirety. PCT / US2020 / 054134, published as WO2021 / 067859(A1), also describes methods for synthesizing such compounds, which are specifically incorporated herein by reference.

[0276] In some embodiments, the HDAC6 inhibitor is a compound of formula (II): [ka] where: n is 0 or 1, X is O, NR 4 , or CR 4 R 4’ and Y is a bond, CR 2 R 3 or S(O)2, R 1 is selected from the group consisting of H, amido, carbocyclyl, heterocyclyl, aryl, and heteroaryl; R 2 and R 3is independently selected from the group consisting of H, halogen, alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH2)-carbocyclyl, -(CH2)-heterocyclyl, -(CH2)-aryl, and -(CH2)-heteroaryl; or R 1 and R 2 together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, or R 2 and R 3 together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, R 4 and R 4’ are each independently selected from the group consisting of H, alkyl, -CO2-alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH2)-carbocyclyl, -(CH2)-heterocyclyl, -(CH2)-aryl, and -(CH2)-heteroaryl; or R 4 and R 4’ together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, Each alkyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is optionally substituted with one or more substituents independently selected from the group consisting of halogen, haloalkyl, oxo, hydroxy, alkoxy, -OCH3, -CO2CH3, -C(O)NH(OH), -CH3, morpholine, and -C(O)N-cyclopropyl.

[0277] Pharmaceutical Compositions and Kits In various embodiments of the present disclosure, a pharmaceutical composition is provided that comprises one or more HDAC6 inhibitors disclosed herein, or a pharma- ceutically acceptable salt thereof, or a pharma- ceutically acceptable solvate, hydrate, tautomer, N-oxide, or salt thereof, and a pharma- ceutically acceptable excipient or adjuvant. Pharmaceutically acceptable excipients and adjuvants are added to compositions or formulations for various purposes. In some embodiments, the pharmaceutical composition that comprises one or more compounds disclosed herein, or a pharma- ceutically acceptable solvate, hydrate, tautomer, N-oxide, or salt thereof, further comprises a pharma- ceutically acceptable carrier. In some embodiments, the pharma- ceutical carrier comprises a pharma- ceutical acceptable excipient, binder, and / or diluent. In some embodiments, suitable pharma- ceutically acceptable excipients include, but are not limited to, water, saline, alcohol, polyethylene glycol (e.g., polyethylene glycol 300), gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, and polyvinylpyrrolidone.

[0278] In some embodiments, the HDAC6 inhibitor in the pharmaceutical compositions described herein is one or more compounds of formula (I), formula (Ia), formula (Ib), formula (Ic), formula (Id), formula (Id-1), formula (Id-2), formula (Id-3), formula (Id-4), formula (Ie), formula (1e-1), formula (If), formula (If-1), formula (Ig), formula (Ig-1), formula (Ih), formula (Ih-1), formula (Ii), formula (Ii-1), formula (Ij), formula (Ij-1), formula (Ik), formula (Ik-1), formula (Ik-2), formula (Ik-3), formula I(y), or formula (II). In some embodiments, the HDAC6 inhibitor in the pharmaceutical compositions described herein is a compound of formula (I). In some embodiments, the HDAC6 inhibitor in the pharmaceutical composition described herein is a compound of formula (Ic). In some embodiments, the HDAC6 inhibitor in the pharmaceutical composition described herein is a compound of formula (Ik). In some embodiments, the HDAC6 inhibitor in the pharmaceutical composition described herein is a compound of formula I(y) (also referred to herein as formula (Iy)).

[0279] In another aspect, the disclosure provides an HDAC6 inhibitor for use in a method for treating a metabolic disease.

[0280] In another aspect, the disclosure provides a kit comprising an HDAC6 inhibitor or a pharmaceutical composition thereof for use in a method for treating a metabolic disease, and instructions.

[0281] In another aspect, the disclosure provides the use of an HDAC6 inhibitor in the treatment of a metabolic disease.

[0282] In another aspect, the disclosure provides an HDAC6 inhibitor for use in a method for treating HFpEF.

[0283] In another aspect, the disclosure provides a kit comprising an HDAC6 inhibitor or a pharmaceutical composition thereof for use in a method for treating HFpEF, and instructions.

[0284] In another aspect, the disclosure provides the use of an HDAC6 inhibitor in the treatment of HFpEF.

[0285] Method of administration The HDAC6 inhibitors described herein (and pharmaceutical compositions comprising such HDAC6 inhibitors) can be administered to a subject by any suitable means disclosed herein or known in the art.

[0286] In some embodiments, the administration of the HDAC6 inhibitor is oral administration. In some embodiments, the method comprises orally administering to the subject an HDAC6 inhibitor of formula (I), formula (Ia), formula (Ib), formula (Ic), formula (Id), formula (Id-1), formula (Id-2), formula (Id-3), formula (Id-4), formula (Ie), formula (1e-1), formula (If), formula (If-1), formula (Ig), formula (Ig-1), formula (Ih), formula (Ih-1), formula (Ii), formula (Ii-1), formula (Ij), formula (Ij-1), formula (Ik), formula (Ik-1), formula (Ik-2), formula (Ik-3), formula I(y) or formula (II). In some embodiments, the method comprises orally administering to the subject an HDAC6 inhibitor of formula (I). In some embodiments, the method comprises orally administering to the subject an HDAC6 inhibitor of formula (Ic). In some embodiments, the method comprises orally administering to the subject an HDAC6 inhibitor of formula (Ik). In some embodiments, the method comprises orally administering to the subject an HDAC6 inhibitor of formula I(y). In some embodiments, the method comprises orally administering to the subject an HDAC6 inhibitor of formula (II). In some embodiments, the oral administration is by tablet or capsule. In some embodiments, a human is orally administered an HDAC6 inhibitor (or a pharmaceutical composition thereof) described herein.

[0287] A variety of administration schedules for the HDAC6 inhibitors described herein (and pharmaceutical compositions containing such HDAC6 inhibitors) are contemplated, including a single administration or multiple administrations over a period of time.

[0288] In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising the inhibitors) are administered twice a day, once a day, once every other day, once every third day, once a week, once every two weeks, once every three weeks, or once a month. In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising the inhibitors) are administered once a day.

[0289] In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising the inhibitors) are administered once. In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising the inhibitors) are administered for a period of time, such as 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year (or more). In some embodiments, the subject being treated is administered the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising the inhibitors) for at least 1 month, at least 6 weeks, at least 2 months, at least 3 months, or at least 6 months. In some embodiments, the subject being treated is administered the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising the inhibitors) for less than 1 month, less than 6 weeks, less than 2 months, less than 3 months, or less than 6 months.

[0290] Appropriate dosages of the HDAC6 inhibitors described herein for use in the methods described herein will depend on the type of inhibitor used, the condition of the subject (e.g., age, weight, health), the subject's responsiveness, other medications used by the subject, and other factors to be considered at the discretion of the medical practitioner administering the treatment.

[0291] In some embodiments, the HDAC6 inhibitors described herein are administered to a subject in an amount ranging from 1 mg to 500 mg per day. In some embodiments, the HDAC6 inhibitors described herein are orally administered to a human in an amount ranging from 1 mg to 500 mg per day. In some embodiments, the HDAC6 inhibitors described herein are orally administered to a human in an amount ranging from 1 mg to 500 mg in a single dose. In some embodiments, the HDAC6 inhibitors described herein are orally administered to a human in an amount ranging from 1 mg to 500 mg once per day, for example, over a course of treatment (e.g., 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, or more). In some embodiments, the HDAC6 inhibitors described herein are administered to a human subject at a starting dose of 0.5 to 3 mg per day (e.g., 1 mg per day). In some embodiments, the HDAC6 inhibitors described herein are administered to human subjects at a starting dose of 0.5 mg / day. In some embodiments, the HDAC6 inhibitors described herein are administered to human subjects at a starting dose of 1 mg / day. In some embodiments, the HDAC6 inhibitors described herein are administered to human subjects at a starting dose of 1.5 mg. In some embodiments, the HDAC6 inhibitors described herein are administered to human subjects at a starting dose of 3 mg. This dose can be titrated up or remain the same depending on the subject's responsiveness to treatment. The HDAC6 inhibitor can be administered in an amount that is considered to be therapeutically effective.

[0292] Combination Treatments and Kits In some embodiments, the HDAC6 inhibitors described herein (and pharmaceutical compositions comprising such HDAC6 inhibitors) can be administered to a subject in combination with another pharmaceutical agent or therapy. In some embodiments, two or three different HDAC6 inhibitors (e.g., among those described herein) can be administered to a subject.

[0293] In some embodiments, one or more of the HDAC6 inhibitors described herein (and pharmaceutical compositions comprising such HDAC6 inhibitors) may be administered to a subject in combination with one or more therapies different from the one or more HDAC6 inhibitors, where the therapy is a cardioprotective therapy, an antihypertensive therapy, a hypoglycemic therapy, a therapy for a cardiac condition (e.g., heart failure), a therapy for HFpEF, or a therapy for a metabolic disease. The additional therapy may be any such therapy known in the art. In some embodiments of metabolic disease treatment, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with another metabolic disease therapy (such as a drug used to treat a metabolic disease). In some embodiments of HFpEF treatment, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with another HFpEF therapy (such as a drug used to treat HFpEF). In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with a hypoglycemic therapy (such as a drug used to treat hypoglycemia). In some embodiments, an HDAC6 inhibitor described herein (or a pharmaceutical composition comprising such an HDAC6 inhibitor) is administered to a subject in combination with an antihypertensive therapy (such as an agent used to treat hypertension).

[0294] In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to subjects in combination with antihypertensive agents. For example, and without limitation, any of the following antihypertensive agents may be used in combination with HDAC6 inhibitors: thiazide diuretics (e.g., Capozide, Dyazide, Hyzaar, Lopressor HCT, Maxzide, Prinzide, Clorpres, Tenoretic, or Thalitone), calcium channel blockers (e.g., Amlodipine, Diltiazem, Felodipine, Isradipine, Nicardipine, Nifedipine, Nisoldipine, or Verapamil), ACE inhibitors (e.g., Benazepril, Captopril, Enalapril, Fosinopril, Lisinopril, Moexipril, Perindopril, l, Quinapril, Ramipril, or Trandolapril), Angiotensin II Receptor Antagonists (ARBs) (e.g., Azilsartan, Candesartan, Eprosartan, Irbesartan, Losartan, Olmesartan, Telmisartan, or Valsartan), and Beta Blockers (e.g., Acebutolol, Atenolol, Bisoprolol, Metoprolol, Nadolol, Nebivolol, or Propranolol).

[0295] In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with an ACE inhibitor. In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with a beta-blocker. In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with a thiazide diuretic. In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with a calcium channel blocker. In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with an angiotensin II receptor antagonist (ARB).

[0296] In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with an aldosterone receptor agonist (e.g., spironolactone).In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with spironolactone.

[0297] In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with an angiotensin and neprilysin receptor agonist (e.g., sacubitril-valsartan). In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with an angiotensin receptor-neprilysin inhibitor (ARNi). In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with sacubitril and / or valsartan.

[0298] In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with a hypoglycemic drug that reduces blood glucose levels (e.g., oral hypoglycemic therapy used to treat type 2 diabetes). For example, and without limitation, any of the following hypoglycemic drugs (or classes of drugs) can be used in combination with the HDAC6 inhibitor: sulfonylureas, meglitinides, biguanides (metformin), thiazolidinediones, alpha-glucosidase inhibitors, DPP4 inhibitors, Cyclotet, and SGLT2 inhibitors. In some embodiments, the HDAC6 inhibitors described herein (or pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with an oral hypoglycemic drug (e.g., used to treat type 2 diabetes).

[0299] In some embodiments, an HDAC6 inhibitor described herein (or a pharmaceutical composition comprising such an HDAC6 inhibitor) is administered to a subject in combination with a sodium-glucose cotransporter 2 (SLGT2) inhibitor (e.g., empagliflozin).

[0300] In some embodiments, the HDAC6 inhibitor described herein (or a pharmaceutical composition comprising such an HDAC6 inhibitor) is administered to the subject before, simultaneously with, or after the additional therapy. In some embodiments, any HDAC6 inhibitor is administered to the subject in a therapeutically effective amount, and any additional therapy described herein is administered to the subject in a therapeutically effective amount.

[0301] In some embodiments, the subject being treated according to the methods described herein is naive to an antihypertensive therapy. In some embodiments, the subject being treated according to the methods described herein is naive to a hypoglycemic therapy. In some embodiments, the subject being treated according to the methods described herein is naive to a metabolic disease therapy. In some embodiments, the subject being treated according to the methods described herein is naive to a HFpEF therapy. In some embodiments, the subject being treated according to the methods described herein is naive to a cardioprotective therapy and / or a cardiac condition (e.g., heart failure) therapy. In some embodiments, the subject being treated according to the methods described herein is naive to an SGLT2 inhibitor.

[0302] In some embodiments, provided herein is a kit comprising an HDAC6 inhibitor (or a pharmaceutical composition comprising the same) described herein and one or more additional agents (e.g., an additional agent for the treatment of a metabolic disease, an additional agent for the treatment of HFpEF, an antihypertensive agent, and / or an antihyperglycemic agent). In some embodiments, provided herein is a kit comprising (i) an HDAC6 inhibitor (e.g., in a therapeutically effective amount) and (ii) one or more additional agents (e.g., any additional agent described herein, including but not limited to a thiazide diuretic, a calcium channel blocker, an ACE inhibitor, an angiotensin II receptor antagonist (ARB), an SGLT2 inhibitor (e.g., empagliflozin), or a beta blocker (e.g., in a therapeutically effective amount).

[0303] Patient population treated In some embodiments, the subject is a human. In some embodiments, the human is an adult human. In some embodiments, the human is 60 years of age or older. In some embodiments, the human is greater than 60 years of age. In some embodiments, the human is 65 years of age or older. In some embodiments, the human is greater than 65 years of age. In some embodiments, the human is 70 years of age or older. In some embodiments, the human is greater than 70 years of age. In some embodiments, the subject is male (e.g., male at least 60, 65, or 70 years of age, or greater than 60, 65, or 70 years of age). In some embodiments, the subject is female (e.g., female at least 60, 65, or 70 years of age, or greater than 60, 65, or 70 years of age).

[0304] In some embodiments, the subject being treated is obese (e.g., has a BMI of 30 or greater). In some embodiments, the subject is at risk for obesity (e.g., has a BMI of 25 or greater). In some embodiments, the subject being treated has a metabolic disease. In some embodiments, the subject is at risk for a metabolic disease. In some embodiments, the subject has diabetes (or is at risk for diabetes). In some embodiments, the subject has diabetes mellitus (or is at risk for diabetes mellitus). In some embodiments, the subject has pre-diabetes (or is at risk for pre-diabetes). In some embodiments, the subject has diabetic cardiomyopathy (or is at risk for diabetic cardiomyopathy). In some embodiments, the subject has metabolic syndrome (or is at risk for metabolic syndrome). In some embodiments, the subject has hypertension (or is at risk for hypertension). In some embodiments, the subject has hypertriglyceridemia (or is at risk for hypertriglyceridemia). In some embodiments, the subject has dyslipidemia (or is at risk for dyslipidemia).

[0305] In some embodiments, the subject being treated is not obese (e.g., has a BMI of less than 30). In some embodiments, the subject has a BMI of less than 25. In some embodiments, the subject being treated does not have a metabolic disease. In some embodiments, the subject being treated does not have diabetes (e.g., does not have diabetes mellitus). In some embodiments, the subject does not have prediabetes. In some embodiments, the subject does not have diabetic cardiomyopathy. In some embodiments, the subject does not have metabolic syndrome. In some embodiments, the subject does not have hypertension. In some embodiments, the subject does not have hypertriglyceridemia. In some embodiments, the subject does not have dyslipidemia. For example, a subject with HFpEF being treated according to the methods described herein may or may not have these conditions.

[0306] In some embodiments, the subject being treated has HFpEF. In some embodiments, the subject being treated is at risk for HFpEF. In some embodiments, the subject has (or is at risk for) cardiac fibrosis. In some embodiments, the subject has (or is at risk for) diastolic dysfunction.

[0307] In some embodiments, the subject has (or is at risk for) coronary artery disease (CAD). In some embodiments, the subject has (or is at risk for) valvular heart disease. In some embodiments, the subject has (or is at risk for) atrial fibrillation. Numbered embodiment (I) of the present invention 1. A method of treating or preventing a metabolic disease in a subject in need thereof, comprising administering to the subject an HDAC6 inhibitor. 2. The method of embodiment 1, wherein the subject is obese or at risk of obesity. 3. The method of embodiment 1 or 2, wherein the metabolic disease is a metabolic disease associated with obesity, optionally diet-induced obesity. 4. The method according to any one of embodiments 1 to 3, wherein the metabolic disease is diabetes, prediabetes, diabetic cardiomyopathy, metabolic syndrome, hypertension, hypertriglyceridemia, or dyslipidemia. 5. The method of embodiment 4, wherein the metabolic disease is diabetes mellitus. 6. The method of embodiment 4, wherein the metabolic disease is metabolic syndrome. 7. A method of treating obesity in a subject in need thereof, comprising administering to the subject an HDAC6 inhibitor. 8. The method of any one of the preceding embodiments, wherein the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative. 9. The method of embodiment 8, wherein the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative according to the following formula: [ka] 10. The HDAC6 inhibitor is a compound according to formula (I): [ka] or a pharma- ceutically acceptable salt thereof, wherein: R 1 is selected from the group consisting of: [Table 15] R a H, halo, C 1-3 selected from the group consisting of alkyl, cycloalkyl, haloalkyl, and alkoxy; R 2 and R 3 is independently selected from the group consisting of H, halogen, alkoxy, haloalkyl, aryl, heteroaryl, alkyl, and cycloalkyl, each of which is optionally substituted, or R 2 and R 3 together with the atom to which they are attached form a cycloalkyl or heterocyclyl; R 4 and R 5 are independently H, -(SO2)R2 , -(SO2)NR 2 R 3 , -(CO)R 2 , -(CONR 2 R 3 ), aryl, arylheteroaryl, alkylenearyl, heteroaryl, cycloalkyl, heterocyclyl, alkyl, haloalkyl, and alkoxy, each of which is optionally substituted; or R 4 and R 5 together with the atom to which they are attached form a cycloalkyl or heterocyclyl, each of which is optionally substituted; R 9 is selected from the group consisting of H, C1-C6 alkyl, haloalkyl, cycloalkyl, and heterocyclyl; X 1 are S, O, NH, and NR 6 R 6 is selected from the group consisting of C1-C6 alkyl, alkoxy, haloalkyl, cycloalkyl, and heterocyclyl; Y is CR 2 , O, N, S, SO, and SO2, and when Y is O, S, SO, or SO2, R 5 does not exist, and R 4 and R 5 When taken together with the atom to which they are attached form a cycloalkyl or heterocyclyl, Y is CR 2 or N, 10. The method of any one of the preceding claims, wherein n is selected from 0, 1, and 2. 11. The method of embodiment 10, wherein the HDAC6 inhibitor is selected from the group consisting of: [Table 16-1] [Table 16-2] [Table 16-3] 12. HDAC6 inhibitors, [ka] 11. The method of embodiment 10, wherein said compound is selected from the group consisting of phenylalanine, ... 13. The method of embodiment 10, wherein the HDAC6 inhibitor is: [ka] 14. The method of embodiment 10, wherein the HDAC6 inhibitor is selected from any HDAC6 inhibitor in Table 13 (Example 7). 15. HDAC6 inhibitors have an IC of 0.03 μM or less 50 15. The method of embodiment 14, wherein the HDAC6 inhibitor is any of Table 13 having the formula: 16. The HDAC6 inhibitor has the formula: [ka] A compound having the formula or a pharma- ceutically acceptable salt thereof; During the ceremony, X 1 is S, R a H, halogens, and C 1-3 is selected from the group consisting of alkyl, R 1 teeth, [ka] and R 2 is selected from the group consisting of alkyl, alkoxy, and cycloalkyl, each of which is optionally substituted; R 3 is H or alkyl, R 4 is alkyl, -(SO2)R 2 , -(SO2)NR 2 R 3 , and -(CO)R 2 is selected from the group consisting of R 5is aryl or heteroaryl, or R 4 and R 5 taken together with the atom to which they are attached form a heterocyclyl, each of which is optionally substituted. 17.R a The method of embodiment 16, wherein 18.R 1 but, [ka] 18. The method of embodiment 16 or 17, wherein 19.R 4 But -(SO2)R 2 19. The method according to any one of embodiments 16 to 18, wherein 20.-(SO2)R 2 The method of embodiment 19, wherein is -(SO2)alkyl, -(SO2)alkyleneheterocyclyl, -(SO2)haloalkyl, -(SO2)haloalkoxy, or -(SO2)cycloalkyl. 21.R 5 The method of any one of embodiments 16-20, wherein is heteroaryl. 22. The method of embodiment 21, wherein the heteroaryl is a 5-6 membered heteroaryl. 23. 5-6 membered heteroaryl is [ka] wherein R b 23. The method of embodiment 22, wherein: is halogen, alkyl, alkoxy, cycloalkyl, -CN, haloalkyl, or haloalkoxy; and m is 0 or 1. 24.R b 24. The method of embodiment 23, wherein is F, Cl, -CH3, -CH2CH3, -CF3, -CHF2, -CF2CH3, -CN, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCF3, -OCHF2, -OCH2CF2H, and cyclopropyl. 25. The method of any one of embodiments 16-24, wherein aryl is selected from the group consisting of phenyl, 3-chlorophenyl, 3-chloro-4-fluorophenyl, 3-trifluoromethylphenyl, 3,4-difluorophenyl, and 2,6-difluorophenyl. 26. The HDAC6 inhibitor has the formula (Ik): [ka] A compound having the formula or a pharma- ceutically acceptable salt thereof; During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 11. The method of embodiment 10, wherein: is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted. 27.R b 27. The method of embodiment 26, wherein is H, halogen, haloalkyl, or haloalkoxy. 28.R 4 28. The method of embodiment 26 or 27, wherein is optionally substituted alkyl or cycloalkyl. 29. An HDAC6 inhibitor having the structure: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 27. The method of embodiment 26, wherein: is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted. 30.R b 30. The method of embodiment 29, wherein is H, halogen, haloalkyl, or haloalkoxy. 31.R 4The method of embodiment 29 or 30, wherein is optionally substituted alkyl or cycloalkyl. 32.R 4 30. The method of embodiment 29, wherein is alkyl. 33. An HDAC6 inhibitor having the structure: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 27. The method of embodiment 26, wherein: is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted. 34.R b is H, halogen, haloalkyl, or haloalkoxy. 35.R 4 The method of embodiment 33 or 34, wherein is optionally substituted alkyl. 36. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 37. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 38. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 39. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 40. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 41. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 42. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 43. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 44. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 45. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 46. ​​The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 47. The HDAC6 inhibitor is a compound of formula (II): [ka] where: n is 0 or 1, X is O, NR 4 , or CR 4 R 4’ and Y is a bond, CR 2 R 3 or S(O)2, R 1 is selected from the group consisting of H, amido, carbocyclyl, heterocyclyl, aryl, and heteroaryl; R 2 and R 3 is independently selected from the group consisting of H, halogen, alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH2)-carbocyclyl, -(CH2)-heterocyclyl, -(CH2)-aryl, and -(CH2)-heteroaryl; or R 1 and R 2 together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, or R 2 and R 3 together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, R 4 and R 4’ are each independently selected from the group consisting of H, alkyl, -CO2-alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH2)-carbocyclyl, -(CH2)-heterocyclyl, -(CH2)-aryl, and -(CH2)-heteroaryl; or R 4 and R 4’ together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, The method of any one of embodiments 1-7, wherein each alkyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently optionally substituted with one or more substituents selected from the group consisting of halogen, haloalkyl, oxo, hydroxy, alkoxy, -OCH3, -CO2CH3, -C(O)NH(OH), -CH3, morpholine, and -C(O)N-cyclopropyl. 48. The method of any one of embodiments 1 to 7, wherein the HDAC6 inhibitor is CAY10603, tubacin, rocilinostat (ACY-1215), citarinostat (ACY-241), ACY-738, QTX-125, CKD-506, nexturastat A, tubastatin A, or HPOB. 49. The method of any one of embodiments 1-48, wherein the compound is a compound and not a pharma- ceutically acceptable salt thereof. 50. The method of any one of embodiments 1-49, wherein the HDAC6 inhibitor is at least 100-fold selective for HDAC6 compared to all other isozymes of HDAC. 51. The method of any one of embodiments 1-50, wherein administration is oral. 52. The method of any one of embodiments 1 to 51, wherein the subject is a human. 53. The method of any one of embodiments 1-52, wherein the subject has or is at risk for hypertension. 54. The method of any one of embodiments 1-53, wherein the subject has or is at risk for diabetes mellitus. 55. The method of any one of embodiments 1-54, wherein the subject has or is at risk for diabetic cardiomyopathy. 56. The method of any one of embodiments 1-55, wherein the subject has or is at risk for metabolic syndrome. 57. The method of any one of embodiments 1-56, wherein the subject has hyperglyceridemia or dyslipidemia or is at risk for hyperglyceridemia or dyslipidemia. 58. The method of any one of embodiments 1 to 57, wherein the subject is a human being who is at least 65 years of age. 59. The method of embodiment 58, wherein the subject is a human being at least 70 years of age. 60. The method of any one of embodiments 1-59, wherein the method treats or prevents at least one symptom of a metabolic disease. 61. The method of any one of embodiments 1-60, wherein the method improves glucose tolerance. 62. The method of any one of embodiments 1-61, wherein the method improves insulin resistance. 63. The method of any one of embodiments 1-62, wherein the method reduces glucose levels. 64. The method of any one of embodiments 1 to 63, wherein the method inhibits inflammatory genes in adipose tissue. 65. The method of any one of embodiments 1-64, wherein the method prevents heart failure in the subject. 66. An HDAC6 inhibitor for use in a method for treating a metabolic disease. 67. The HDAC6 inhibitor according to embodiment 66, wherein the HDAC6 inhibitor is any one of those described in embodiments 8 to 50. 68. A pharmaceutical composition for use in a method for treating a metabolic disease, comprising an HDAC6 inhibitor. 69. The pharmaceutical composition according to embodiment 68, wherein the HDAC6 inhibitor is any one of those described in embodiments 8 to 50. 70. A kit comprising an HDAC6 inhibitor and instructions for use in a method for treating a metabolic disease. 71. The kit according to embodiment 70, wherein the HDAC6 inhibitor is any one of those described in embodiments 8 to 50. 72. Use of HDAC6 inhibitors in the treatment of metabolic diseases. 73. The use according to embodiment 72, wherein the HDAC6 inhibitor is any one of those described in embodiments 8 to 50. Numbered embodiment (II) of the present invention 1. A method of treating or preventing heart failure with preserved ejection fraction (HFpEF) in a subject in need thereof, comprising administering to the subject an HDAC6 inhibitor. 2. A method of treating or preventing diastolic dysfunction (e.g., associated with HFpEF) in a subject in need thereof, comprising administering an HDAC6 inhibitor to the subject. 3. A method of treating or preventing cardiac fibrosis (e.g., associated with HFpEF) in a subject in need thereof, comprising administering to the subject an HDAC6 inhibitor. 4. The method of any one of embodiments 1-3, wherein administration is oral. 5. The method of any one of embodiments 1 to 4, wherein the subject is a human. 6. The method of embodiment 5, wherein the subject is at least 65 years of age. 7. The method of embodiment 5, wherein the subject is at least 70 years of age. 8. The method of any one of the preceding embodiments, wherein the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative. 9. The method of embodiment 8, wherein the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative according to the following formula: [ka] 10. The HDAC6 inhibitor is a compound according to formula (I): [ka] or a pharma- ceutically acceptable salt thereof, wherein: R 1 is selected from the group consisting of: [Table 17] R a H, halo, C 1-3selected from the group consisting of alkyl, cycloalkyl, haloalkyl, and alkoxy; R 2 and R 3 is independently selected from the group consisting of H, halogen, alkoxy, haloalkyl, aryl, heteroaryl, alkyl, and cycloalkyl, each of which is optionally substituted, or R 2 and R 3 together with the atom to which they are attached form a cycloalkyl or heterocyclyl; R 4 and R 5 are independently H, -(SO2)R 2 , -(SO2)NR 2 R 3 , -(CO)R 2 , -(CONR 2 R 3 ), aryl, arylheteroaryl, alkylenearyl, heteroaryl, cycloalkyl, heterocyclyl, alkyl, haloalkyl, and alkoxy, each of which is optionally substituted; or R 4 and R 5 together with the atom to which they are attached form a cycloalkyl or heterocyclyl, each of which is optionally substituted; R 9 is selected from the group consisting of H, C1-C6 alkyl, haloalkyl, cycloalkyl, and heterocyclyl; X 1 are S, O, NH, and NR 6 R 6 is selected from the group consisting of C1-C6 alkyl, alkoxy, haloalkyl, cycloalkyl, and heterocyclyl; Y is CR 2 , O, N, S, SO, and SO2, and when Y is O, S, SO, or SO2, R 5 does not exist, and R 4 and R 5 When taken together with the atom to which they are attached form a cycloalkyl or heterocyclyl, Y is CR2 or N, 10. The method of any one of the preceding claims, wherein n is selected from 0, 1, and 2. 11. The method of embodiment 10, wherein the HDAC6 inhibitor is selected from the group consisting of: [Table 18-1] [Table 18-2] [Table 18-3] 12. HDAC6 inhibitors, [ka] 11. The method of embodiment 10, wherein said compound is selected from the group consisting of phenylalanine, ... 13. The method of embodiment 10, wherein the HDAC6 inhibitor is: [ka] 14. The method of embodiment 10, wherein the HDAC6 inhibitor is selected from any HDAC6 inhibitor in Table 13 (Example 7). 15. HDAC6 inhibitors have an IC of 0.03 μM or less 50 15. The method of embodiment 14, wherein the HDAC6 inhibitor is any of Table 13 having the formula: 16. The HDAC6 inhibitor has the formula: [ka] A compound having the formula or a pharma- ceutically acceptable salt thereof; During the ceremony, X 1 is S, R a H, halogens, and C 1-3 is selected from the group consisting of alkyl, R 1 teeth, [ka] and R 2 is selected from the group consisting of alkyl, alkoxy, and cycloalkyl, each of which is optionally substituted; R 3 is H or alkyl, R 4 is alkyl, -(SO2)R 2 , -(SO2)NR 2 R 3 , and -(CO)R 2 is selected from the group consisting of R 5 is aryl or heteroaryl, or R 4 and R 5 taken together with the atom to which they are attached form a heterocyclyl, each of which is optionally substituted. 17.R a The method of embodiment 16, wherein 18.R 1 but, [ka] 18. The method of embodiment 16 or 17, wherein 19.R 4 But -(SO2)R 2 19. The method according to any one of embodiments 16 to 18, wherein 20.-(SO2)R 2 The method of embodiment 19, wherein is -(SO2)alkyl, -(SO2)alkyleneheterocyclyl, -(SO2)haloalkyl, -(SO2)haloalkoxy, or -(SO2)cycloalkyl. 21.R 5 The method of any one of embodiments 16-20, wherein is heteroaryl. 22. The method of embodiment 21, wherein the heteroaryl is a 5-6 membered heteroaryl. 23. 5-6 membered heteroaryl is [ka] wherein R b 23. The method of embodiment 22, wherein: is halogen, alkyl, alkoxy, cycloalkyl, -CN, haloalkyl, or haloalkoxy; and m is 0 or 1. 24.R b 24. The method of embodiment 23, wherein is F, Cl, -CH3, -CH2CH3, -CF3, -CHF2, -CF2CH3, -CN, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCF3, -OCHF2, -OCH2CF2H, and cyclopropyl. 25. The method of any one of embodiments 16-24, wherein aryl is selected from the group consisting of phenyl, 3-chlorophenyl, 3-chloro-4-fluorophenyl, 3-trifluoromethylphenyl, 3,4-difluorophenyl, and 2,6-difluorophenyl. 26. The HDAC6 inhibitor has the formula (Ik): [ka] A compound having the formula or a pharma- ceutically acceptable salt thereof; During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 11. The method of embodiment 10, wherein: is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted. 27.R b 27. The method of embodiment 26, wherein is H, halogen, haloalkyl, or haloalkoxy. 28.R 4 28. The method of embodiment 26 or 27, wherein is optionally substituted alkyl or cycloalkyl. 29. An HDAC6 inhibitor having the structure: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 27. The method of embodiment 26, wherein: is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted. 30.R b 30. The method of embodiment 29, wherein is H, halogen, haloalkyl, or haloalkoxy. 31.R 4 The method of embodiment 29 or 30, wherein is optionally substituted alkyl or cycloalkyl. 32.R 4 30. The method of embodiment 29, wherein is alkyl. 33. An HDAC6 inhibitor having the structure: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, R b is H, halogen, alkyl, cycloalkyl, -CN, haloalkyl, or haloalkoxy; R 4 27. The method of embodiment 26, wherein: is alkyl, alkoxy, haloalkyl, or cycloalkyl, each of which is optionally substituted. 34.R b is H, halogen, haloalkyl, or haloalkoxy. 35.R 4 The method of embodiment 33 or 34, wherein is optionally substituted alkyl. 36. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 37. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 38. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 39. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 40. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 41. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 42. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 43. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 44. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 45. The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 46. ​​The HDAC6 inhibitor has the formula: [ka] or a pharma- ceutically acceptable salt thereof. 47. The HDAC6 inhibitor is a compound of formula (II): [ka] where: n is 0 or 1, X is O, NR 4 , or CR 4 R 4’ and Y is a bond, CR 2 R 3 or S(O)2, R 1 is selected from the group consisting of H, amido, carbocyclyl, heterocyclyl, aryl, and heteroaryl; R 2 and R 3 is independently selected from the group consisting of H, halogen, alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH2)-carbocyclyl, -(CH2)-heterocyclyl, -(CH2)-aryl, and -(CH2)-heteroaryl; or R 1 and R 2 together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, or R 2 and R 3 together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, R 4 and R 4’ are each independently selected from the group consisting of H, alkyl, -CO2-alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH2)-carbocyclyl, -(CH2)-heterocyclyl, -(CH2)-aryl, and -(CH2)-heteroaryl; or R 4 and R 4’ together with the carbon atom to which they are attached form a carbocyclyl or heterocyclyl, The method of any one of embodiments 1-7, wherein each alkyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently optionally substituted with one or more substituents selected from the group consisting of halogen, haloalkyl, oxo, hydroxy, alkoxy, -OCH3, -CO2CH3, -C(O)NH(OH), -CH3, morpholine, and -C(O)N-cyclopropyl. 48. The method of any one of embodiments 1 to 7, wherein the HDAC6 inhibitor is CAY10603, tubacin, rocilinostat (ACY-1215), citarinostat (ACY-241), ACY-738, QTX-125, CKD-506, nexturastat A, tubastatin A, or HPOB. 49. The method of any one of embodiments 1-48, wherein the compound is a compound and not a pharma- ceutically acceptable salt thereof. 50. The method of any one of embodiments 1-49, wherein the HDAC6 inhibitor is at least 100-fold selective for HDAC6 compared to all other isozymes of HDAC. 51. The method of any one of embodiments 1-50, wherein the subject has or is at risk for hypertension. 52. The method of any one of embodiments 1-51, wherein the subject has or is at risk for diabetes mellitus. 53. The method of any one of embodiments 1-52, wherein the subject has or is at risk for coronary artery disease (CAD). 54. The method of any one of embodiments 1-53, wherein the subject has or is at risk for valvular heart disease. 55. The method of any one of embodiments 1-54, wherein the subject has atrial fibrillation or is at risk for atrial fibrillation. 56. The method of any one of embodiments 1-55, wherein the method treats or prevents at least one symptom of HFpEF. 57. The method of any one of embodiments 1-56, wherein the method reduces left ventricular (LV) mass. 58. The method of any one of embodiments 1 to 57, wherein the method reduces LV wall thickness. 59. The method of any one of embodiments 1 to 58, wherein the method improves LV relaxation. 60. The method of any one of embodiments 1-59, wherein the method improves LV filling pressure. 61. The method of any one of embodiments 1-60, wherein the method prevents heart failure in a subject. 62. An HDAC6 inhibitor for use in a method for treating heart failure with preserved ejection fraction. 63 The HDAC6 inhibitor of embodiment 62, wherein the HDAC6 inhibitor is any one of embodiments 8 to 50. 64. A pharmaceutical composition for use in a method for treating heart failure with preserved ejection fraction, comprising an HDAC6 inhibitor. 65. The pharmaceutical composition according to embodiment 64, wherein the HDAC6 inhibitor is any one of those described in embodiments 8 to 50. 66. A kit comprising an HDAC6 inhibitor and instructions for use in a method for treating heart failure with preserved ejection fraction. 67. The kit according to embodiment 66, wherein the HDAC6 inhibitor is any one of those described in embodiments 8 to 50. 68. Use of HDAC6 inhibitors in the treatment of heart failure with preserved ejection fraction. 69. The use according to embodiment 68, wherein the HDAC6 inhibitor is any one of those described in embodiments 8 to 50. EXAMPLES

[0308] The present invention is further illustrated by the following examples, which are non-limiting and merely representative of various aspects of the present invention.

[0309] Example 1: HDAC6 selective inhibitors improve glucose and insulin resistance in a diet-induced obese mouse model This example demonstrates that histone deacetylase 6 (HDAC6) selective inhibitors improve glucose tolerance and insulin resistance in a diet-induced obesity mouse model. The diet-induced obesity model closely mimics the high fat / high density diet that contributes to obesity in humans. (Reviewed in Wang et al. Methods Mol Biol. 821:421-433 (2012)).

[0310] 1A-1L show that HDAC6 inhibition with TYA-11631 improves glucose tolerance and insulin resistance in a diet-induced obese mouse model. TYA-11631 is a compound within formula (I), as well as compounds within formula (Ic), formula (Ik), and formula I(y).

[0311] Sixteen week old male diet-induced obese (DIO) C57BL / 6J (cat. 380050) mice and age / sex matched controls (cat. 000664) were purchased from Jackson Laboratory®. Starting at 6 weeks of age, DIO mice (n=40) were fed a rodent diet containing 60 kcal% fat (D12492) and control mice were fed a rodent diet containing 10 kcal% fat (D12450B).

[0312] Intraperitoneal glucose tolerance test (GTT) was performed by injection of glucose (2 g / kg in saline) after 6 h fasting. Tail blood glucose levels (mg / dl) were measured with a glucometer before glucose administration (0 min) and 15, 30, 45, 60, and 120 min after glucose administration. 16-week-old DIO mice developed severe glucose intolerance compared to controls (Figure 1A, Figure 1B).

[0313] Based on glucose AUC levels, DIO mice were randomized equally into four treatment groups and administered vehicle (n=9) or TYA-11631 at three doses: 3, 10, 30 mg / kg (n=10 each). Control mice were also split to be orally administered vehicle (n=10) or 30 mg / kg TYA-11631 (n=10). To evaluate the acute response of TYA-11631 on glucose metabolism, a GTT was performed 6 hours after the first dose. Surprisingly, a single dose of TYA-11631 at all three tested doses significantly reduced glucose levels. (Figure 1C, Figure 1D). Two weeks of TYA-11631 treatment resulted in a significant improvement in glucose tolerance in a dose-dependent manner. (Figure 1E, Figure 1F).

[0314] To evaluate the effect of TYA-11631 on insulin resistance, intraperitoneal insulin tolerance test (ITT) was performed by injection of insulin (1 U / kg) after 6 hours of fasting. Tail blood glucose levels (mg / dl) were measured with a glucometer before insulin administration (0 min) and 15, 30, 45, 60, and 120 min after insulin administration. Four weeks of HDAC6 inhibitor (TYA-11631) treatment improved insulin resistance in DIO mice. 10 and 30 mg / kg significantly reduced glucose AUC (ITT) with comparable activity, and 3 mg / kg showed a trend toward reduction. (Figure 1G, Figure 1H)

[0315] The effect of TYA-11631 on blood glucose in non-fasting mice was also evaluated. Tail blood samples were collected in the morning and measured with a glucometer. TYA-11631 treatment for 6 weeks resulted in a dose-dependent reduction in non-fasting glucose, consistent with the data from the glucose tolerance test after fasting. (Figure 1I)

[0316] Treatment with TYA-11631 caused a dose-dependent reduction in body weight in DIO mice (see Fig. 1J, K, L). No difference in food intake was observed between groups. Of note, control mice administered TYA-11631 30 mg / kg for 6 weeks showed no changes in blood glucose levels and body weight. Bars and error bars indicate mean and SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0317] 2A-C show that HDAC6 inhibition with TYA-11631 inhibits inflammatory genes in adipose tissue of a diet-induced obesity mouse model.

[0318] White adipose tissue (epididymis) was excised from DIO mice 6 hours after a single dose of 30 mg / kg TYA-11631. Real-time q-PCR data showed that TYA-11631 significantly inhibited the upregulation of pro-inflammatory genes-IL-6 (Figure 2A), IL-10 (Figure 2B), and TGFb1 (Figure 2C) in white adipose tissue. Inhibition or ablation of these genes has been shown to protect mice from diet-induced obesity, accompanied by improved glucose tolerance and insulin sensitivity (Han et al., PNAS 2020; Rajbhandari et al., Cell 2018).

[0319] Taken together, these data show that histone deacetylase 6 (HDAC6) selective inhibitors improve glucose tolerance and insulin resistance in a diet-induced obese mouse model and suggest potential molecular mechanisms underlying the effects on whole-body glucose metabolism.

[0320] Example 2: HDAC6 inhibitors improve glucose metabolism and symptoms of metabolic disease in diabetic mouse models This example demonstrates that a histone deacetylase 6 (HDAC6) selective inhibitor ameliorates glucose metabolism and symptoms of metabolic disease in a mouse model of diabetes (independent of high-fat diet).

[0321] Materials and Methods TYA-11018, an HDAC6 inhibitor, was used in this example. TYA-11018 is a compound within formula (I), and for example, compounds within formula (Ic) and formula I(y). Vehicle contained 5% DMSO+45% PEG-300 in purified water.

[0322] Test system: An 8-week-old male db / db (cat. 000697, homozygous Lepr db ) mice and age / sex-matched wild-type controls (catalog 000664) were purchased from Jackson Laboratory (Bar Harbor, ME). Animals were fed a normal chow diet (LabDiet 5053, 10 kcal% fat diet, St Louis, MO) and housed in an animal research facility in accordance with the National Research Council of the National Academies guidelines for the care and use of laboratory animals. All mice were used in the experiments after a one-week acclimation period.

[0323] Baseline Glucose and Animal Randomization: Fed state blood samples (before administration of TYA-11018 or vehicle alone) were taken from the tail vein to determine baseline glucose. Based on both body weight and blood glucose levels, db / db mice were randomized into three groups and orally administered vehicle or TYA-11018 at two doses of 1.5 or 15 mg / kg. Wild-type mice were orally administered vehicle for control.

[0324] Single oral dose and blood glucose test: After a single dose of TYA-11018 or vehicle was administered via stomach tube at a volume of 10 ml / kg, the animals were transferred to clean cages and wired to a water bottle at the top for fasting. Fasting blood samples were collected from the tail vein at 0.5, 1, 2, 4, and 6 hours after the single dose of TYA-11018 or vehicle for glucose determination. Blood glucose levels (mg / dL) were measured by AimStrip Plus blood glucose strips used in the AimStrip Plus blood glucose monitoring system (Germaine Laboratories, San Antonio, TX).

[0325] result Animal Enrollment and Randomization: Animals were randomized and enrolled in the single-dose study based on body weight and baseline glucose. Body weight and tail blood glucose levels (mg / dL) were measured in the fed state immediately prior to TYA-11018 or vehicle administration. Nine-week-old db / db mice developed severe obesity with hyperglycemia compared to controls. Animals were randomized and distributed equally into three groups to receive vehicle (n=8) or two doses of TYA-11018, 1.5 (n=8) and 15 mg / kg (n=9). Controls (age- and sex-matched wild-type mice) were orally administered vehicle (n=8) (Table). [Table 3]

[0326] A single dose of TYA-11018 dose-dependently reduced fasting glucose in db / db diabetic mice: To evaluate the acute response of TYA-11018 on glucose metabolism, blood glucose was measured 0.5, 1, 2, 4, and 6 hours after a single dose. As shown in Figures 3A-3C, treatment with TYA-11018 showed a dose-dependent effect on reducing fasting glucose. After 4 hours of dosing / fasting, both 1.5 and 15 mg / kg significantly reduced blood glucose compared to vehicle-treated db / db diabetic mice. Notably, a single dose of 15 mg / kg TYA-11018 normalized fasting glucose at 4 hours. These data support the utility of HDAC6 selective inhibitors in improving glucose metabolism and symptoms of metabolic disease.

[0327] Example 3: HDAC6 selective inhibitors improve cardiac structure and function in HFpEF mouse models This example demonstrates that a histone deacetylase 6 (HDAC6) selective inhibitor improves cardiac structure and function in a mouse model of heart failure with preserved ejection fraction (HFpEF).

[0328] First, we established an animal model of HFpEF: we found that surgically applying moderate transverse aortic constriction (mTAC) in wild-type C57BL6 mice fed a high-fat diet (HFD) induces a cardiometabolic heart failure phenotype that recapitulates the systemic and cardiovascular features of HFpEF in humans (Figures 4A-4H).

[0329] HFpEF model animals were orally administered 30 mg / kg of HDAC6 inhibitor (TYA-11631) or vehicle once daily for 6 weeks. HDAC6 inhibitors effectively treated HFpEF. HDAC6 inhibitor treatment regimens significantly reduced left ventricular (LV) mass (Figure 5E) and LV wall thickness (Figure 5F) measured by echocardiogram, and HDAC6 inhibitor treatment improved glucose tolerance (Figure 5A). Furthermore, TYA-11631 treatment improved LV relaxation and LV filling pressures, as indicated by a reduction in isovolumic relaxation time prolongation (Figure 5H), reduced E / A (Figure 5I) and E / e' ratios (Figure 5L), improved e' velocity (Figure 5K), and reduced end-diastolic pressure (Figure 5M). Each of these efficacy parameters was normalized to control levels.

[0330] Furthermore, HFD / mTAC mice treated with TYA-11631 showed a trend toward decreased lung weight (Figure 5O), indicating amelioration of pulmonary congestion, consistent with reduced filling pressures. No treatment-related adverse events or toxicity were observed in the animals in this study.

[0331] At the molecular level, HDAC6 inhibitors significantly inhibited the upregulation of genes commonly associated with fibrosis (Postn, Col1a1, Col3a1, Col5a2), cardiac stress (Nppb), and inflammation (Tnfα) in cardiac tissue from HFD / mTAC mice (Figures 6A-6H).

[0332] The results demonstrate that selectively inhibiting HDAC6 reverses multiple deleterious pathophysiological processes in HFpEF.

[0333] Example 4: HDAC6-selective inhibitors improve cardiac structure and function in a second HFpEF mouse model This example demonstrates that a histone deacetylase 6 (HDAC6) selective inhibitor improves cardiac structure and function in a second mouse model of heart failure with preserved ejection fraction (HFpEF) that recapitulates most of the clinical features of the syndrome: high-fat diet (HFD) with inhibition of constitutive nitric oxide synthase (NOS) using N[w]-nitro-l-arginine methyl ester (L-NAME) (Schiattarella et al. Nature 568(7752):351-356(2019)).

[0334] First, we validated the mouse model. HFD / L-NAME treatment significantly induced weight gain (Figure 7B), hypertension (Figure 7C), and glucose intolerance (Figure 7D) compared with control mice. Echocardiographic evaluation revealed sustained preservation of left ventricular ejection fraction (LVEF) (Figure 7E). Significant concentric left ventricular (LV) hypertrophy was present in HFD / L-NAME mice, as shown by increased LV mass (Figure 7F) and LV wall thickness (Figure 7G) in diastole, without LV cavity dilation (Figure 7H). Furthermore, mice simultaneously exposed to HFD / L-NAME showed signs of LV diastolic dysfunction with impaired relaxation and increased LV filling pressure, as evidenced by prolonged IVRT (Figure 7I), decreased e' velocity (Figure 7J), and increased E / e' (Figure 7K), E / A (Figure 7L) ratios, measured by noninvasive Doppler imaging.

[0335] Second, treatment with the HDAC6 inhibitor, TYA-11631, was shown to improve glucose tolerance and diastolic dysfunction in the HFD / L-NAME model.

[0336] Animals were randomized to receive oral administration of 30 mg / kg TYA-11631 (n=8) or vehicle (n=12) once daily for 9 weeks. In the control group, 6 mice received vehicle and 8 mice received 30 mg / kg TYA-11631. Glucose tolerance tests (GTTs) were performed after 5 weeks of treatment, as shown in Figures 8A and 8B.

[0337] Treatment with TYA-11631 significantly improved glucose tolerance in HFD / L-NAME mice, while there was no change in control animals. (Figure 8B) Plasma insulin levels during GTT at the indicated time points (0 and 30 min after glucose injection) were measured by a highly sensitive mouse insulin detection kit (catalog 80-INSMS-E01, ALPCO). TYA-11631 treatment led to a decrease in insulin secretion, suggesting that the improvement in glucose tolerance may be due to improved insulin action / sensitivity. (Figure 8C) Treatment with TYA-11631 caused a significant reduction in body weight, but no difference in food intake, in mice fed HFD / L-NAME. (Figure 8D, Figure 8E) TYA-11631 did not affect systolic blood pressure in HFD / L-NAME mice measured by the non-invasive tail cuff method. (FIG. 5F) Echocardiography showed that TYA-11631 treatment preserved ejection fraction (FIG. 8G), but significantly reduced left ventricular mass (FIG. 8H) and LV wall thickness (FIG. 8I).

[0338] Noninvasive Doppler imaging and distal invasive catheterization analysis revealed that treatment with TYA-11631 for 9 weeks reduced isovolumic relaxation time prolongation (Figure 8J), E / A ratio (Figure 8K), and E / e' ratio (Figure 8L), increased e' velocity (Figure 8M), and reduced end-diastolic pressure (Figure 8N), indicating improved LV relaxation and LV filling pressures. Furthermore, HFD / L-NAME mice treated with TYA-11631 showed a trend toward reduced lung weight (Figure 8O), suggesting improved pulmonary congestion and consistent with reduced filling pressures.

[0339] Of note, no adverse effects associated with TYA-11631 were observed: control animals administered TYA-11631 had no changes in each of the LV structural and functional parameters, as well as ECG signals-QT, QRS, PR intervals, and R amplitudes (Figures 8P, 8Q, 8R, and 8S), further supporting the overall favorable safety profile of the compound.

[0340] Example 5: HDAC6 inhibitors reduce cardiac fibrosis and enhance mitochondrial function in a mouse model of HFpEF This example demonstrates that histone deacetylase 6 (HDAC6) selective inhibitors reduce cardiac fibrosis and enhance mitochondrial function in a mouse model of HFpEF. Specifically, this example demonstrates that HDAC6 selective inhibitors (i) correct dysregulated fibrosis and oxidative phosphorylation gene expression in HFpEF models, (ii) increase mitochondrial membrane potential and spare respiratory capacity in human iPSC-CMs, and (iii) prevent fibroblast activation from TGF-beta in primary human cardiac fibroblasts.

[0341] Transcriptional analysis was performed on cardiac tissue from mice with HFpEF using RNA sequencing and qPCR, as described below. The RNA-Seq data presented herein shows reduced expression of gene sets related to hypertrophy, fibrosis, and PDGFR signaling in HFpEF mice treated with TYA-11631. qPCR data further confirmed the reduced expression of fibrotic genes correlated with improved diastolic function. Furthermore, the RNA-Seq data presented herein shows that gene sets related to mitochondrial energy production were enriched in TYA-11631-treated HFpEF mice. Increased expression of mitochondrial genes correlated with improved diastolic function. TYA-11631 was further tested to determine whether it has a direct effect on metabolism in an in vitro model using human induced pluripotent stem cell-derived cardiomyocytes. The data presented herein show enhanced spare respiratory capacity and improved ATP production in response to stress in TYA-11631-treated human iPSC-derived CMs.

[0342] Materials and Methods HFpEF was surgically induced by 12 weeks of moderate transverse aortic constriction in mice fed a high-fat diet (mTAC / HFD). After the HFpEF phenotype was established, mice were orally administered TYA-11631 (30 mg / kg) or vehicle once daily for 6 weeks. TYA-11631-treated mice showed improved cardiac function as measured by reductions in left ventricular posterior wall thickness (LVPWd), isovolumic relaxation time (IVRT), and mitral valve E / e' (MV E / e'). TYA-11631 is a compound within formula (I), as well as compounds within formula (Ic), formula (Ik), and formula I(y).

[0343] Testing Systems Twelve-week-old male C57Bl / 6NJ mice were purchased from Jackson Laboratories. Mice were maintained under specific pathogen-free conditions and provided with sterile food and water ad libitum. Animals were housed in an animal research facility in accordance with the National Research Council of the National Academies guidelines for the care and use of laboratory animals. Animals were allowed to acclimate for 3 days prior to experimentation.

[0344] Induced pluripotent stem cell-derived cardiomyocytes (iPSC-derived cardiomyocytes): iCell Cardiomyocyte 2 was purchased from Cellular Dynamics (Madison, WI). Cells were cultured in low glucose and lipid-enriched medium to enhance cell maturation for one week prior to treatment.

[0345] RNA extraction and mRNA sequencing (RNA-Seq) methods and analysis RNA was extracted from mouse heart tissue (control mice, vehicle-treated, n=3; HFpEF mice, vehicle-treated, n=5; and HFpEF mice, TYA-11631-treated, n=6) using a polyA tail-specific protocol from Illumina (San Diego, CA, catalog no. 20020594). RNA library preparation and ribosomal RNA removal were performed on 100 ng of RNA using the TruSeq Stranded mRNA kit (Illumina, San Diego, CA, catalog no. 20020594) according to the manufacturer's instructions. Libraries were sequenced using Illumina NovaSeq SP as 100 bp single-end reads with an average of 45.7 million reads per sample. Raw RNA-seq reads in FASTQ format were directly aligned to GENCODE (version M25) for reference transcript assemblies (GRCm38.p6 and ensemble 101). A script using tximport was then used to generate an expression matrix normalized to transcripts per million (TPM). Only genes detected in at least 90% of all samples were used in this analysis. Protein-coding genes were determined using the Ensembl release Mus musculus annotation (GRCm38, April 2020) and extracted by biomaRt (version 2.46.3). Non-protein-coding and mitochondrial genes were omitted and subsequently re-normalized to TPM. The generated expression matrix (16,499 genes) was log2 transformed after adding 1 as a pseudocount.

[0346] To assess functional perturbations, a pre-ranked gene set enrichment analysis was performed using GSEA (version 4.1.0) developed by the Broad institute. GSEA assesses whether the difference in expression of a gene set between two phenotypes is statistically significant (Subramanian, 2005). Prior to the analysis, a ranked list was calculated and each gene was assigned a score and direction ('+' or '-') based on the t-statistic. Gene sets were considered statistically significant only if the false discovery rate (FDR) was less than 0.25 as determined by the multiple hypothesis testing correction method (Benjamini, 1995). The normalized enrichment score (NES), which reflects the degree to which a gene set is over-represented in the ranked list and normalized for gene set size, was used to select significantly altered gene sets. Correlations between genes of interest and cardiac diastolic function parameters were calculated using Pearson correlation coefficients.

[0347] RNA extraction and TaqMan qPCR analysis methods 15-30 mg of mouse heart tissue (control mice, vehicle-treated, n=3; HFpEF mice, vehicle-treated, n=5; and HFpEF mice, TYA-11631-treated, n=6) was placed directly into 500 μL of Tri-Reagent (Zymo research, Irvine, CA, Cat. No. R2050-1-200) and flash frozen at -80°C. Samples were thawed and homogenized by Bullet Blender Tissue Homogenizer (Storm Pro BT24M, Next Advance, Troy) for 15 min at 4°C. RNA was extracted using Direct-Zol RNA Miniprep Plus Kit (Zymo research, Irvine, CA, Cat. No. R2070) according to the manufacturer's instructions. After RNA was harvested, the concentration was determined by NanoDrop (ThermoScientific, Waltham, MA,). cDNA was reverse transcribed from 750 ng of RNA through random hexamers using the SuperScriptIII kit (Invitrogen, Waltham, MA, Catalog No. 18080051). cDNA samples were diluted 6-fold in nuclease-free water. Real-time qPCR reactions were performed using standard TaqMan Universal PCR Master Mix (Applied Biosystems, Waltham, MA, Catalog No. 43-181-57) with the TaqMan probes (Life Technologies, San Diego, CA) listed in Table 4. [Table 4]

[0348] Real-time qPCR reactions were performed using QuantStudio7 Flex Real-Time PCR Systems (Life Technologies, San Diego, CA) with thermal cycling parameters of 2 min UNG incubation at 50°C, 10 min polymerase activation at 95°C, and 40 PCR cycles consisting of 15 s denaturation at 95°C and 1 min annealing / extension at 60°C. Gene expression was normalized to Gapdh as a housekeeping gene. Four technical replicates were analyzed for each sample. -ΔΔC T Relative gene expression was determined using the FTIR method. Statistical analysis was performed with Prism Version 9 (GraphPad Software, San Diego, CA) using unpaired t-tests.

[0349] Metabolic measurements using Seahorse Oximetry The metabolic activity of human induced pluripotent stem cell-derived cardiomyocytes (iPSC-derived CMs) was assessed using a Seahorse XF96 Analyzer (Agilent, Santa Clara, CA), which measures the oxygen consumption rate (OCR) of live cells in multiwell plates. Seahorse XF96 V3 PS Cell Culture Microplates (catalog no. 101085-004) were coated overnight with 100 μL of Matrigel (Corning, Corning, NY, catalog no. 356231) at a 1 / 100 dilution in phenol-free DMEM medium. The next day, Matrigel was removed and 25 μL of Seeding Medium (Cellular Dynamics, Madison, WI, catalog no. R1151) was added to each well of the Seahorse XF96 V3 PS Cell Culture Microplates. iCell Cardiomyocytes 2One vial of iPSC-derived CMs (approximately 6 million cells, Cellular Dynamics, Madison, WI, catalog number 01434) was thawed for 2 minutes in a 37°C water bath. Cells were seeded directly onto Seahorse XF96 V3 PS Cell Culture Microplates for a final density of 15,000 cells per well in seeding medium. Corner wells were excluded for background recording. Cells were left at room temperature for 10 minutes to evenly distribute the seeding and then placed in a 37°C incubator. Five hours after cell seeding, the medium was changed to Maintenance Medium (Cellular Dynamics, Madison, WI, catalog number R1151) for three days for recovery. Once a beating monolayer was observed, the medium was changed to Fatty Acid Enriched Maturation Medium (Feyen, 2020) for one week to increase the metabolic maturity of iPSC-derived CMs. Cells were replenished with fresh medium every three days. On the day of the assay, cells were incubated for 1 h in starvation medium containing 2 mM glutamine in DMEM (Agilent, Santa Clara, CA, Cat. No. 103575-100). Cells were then treated with DMSO (0.1%) or TYA-11631 (3 μM) in Mercola medium for 6 h. Cells were washed and incubated for 1 h before assay with Seahorse XF DMEM Basal Medium supplemented with 2 mM glutamine, 2 mM pyruvate, and 3 mM glucose. Seahorse Xfe96 cartridges were prepared according to the manufacturer's guidelines. Oxygen consumption rate (OCR) was measured, followed by a Mito Stress Test (Agilent, Santa Clara, CA, Cat. No. 103015-100) with inhibitors injected in the following order: oligomycin (2.5 μM), FCCP (1 μM), rotenone, and antimycin A (0.5 μM). OCR was normalized to total nuclei number as measured by Hoechst staining. Basal respiration was calculated as the last rate measurement before the first oligomycin injection minus the minimum rate measurement after rotenone / antimycin. Reserve respiratory capacity (RRC) was calculated as the maximum respiration after addition of FFCP minus basal respiration.Statistical analysis was performed with Prism Version 9 (GraphPad Software, San Diego, CA) using unpaired t tests.

[0350] result Transcriptional analysis of TYA-11631 effects on HFpEF mouse models by RNA-Seq Global transcriptional profiling of protein-coding genes in cardiac tissue from control and HFpEF mice (mTAC / HFD) was performed using RNA sequencing. To assess functional perturbations, unbiased gene set enrichment analysis was performed in two comparisons: 1) vehicle-treated HFpEF mouse tissue compared to vehicle-treated control mouse tissue, and 2) TYA-11631-treated compared to vehicle-treated HFpEF mouse tissue.

[0351] Gene sets related to muscle hypertrophy and contraction, fibrosis (transforming growth factor beta receptor signaling, type I collagen synthesis, extracellular matrix structural components), and platelet-derived growth factor receptor (PDGFR) signaling were enriched in vehicle-treated HFpEF animals compared to controls and reversed in HFpEF mice treated with TYA-11631 (Figure 9 and Error! Reference source not found.0). Gene sets related to mitochondrial function, including electron transport chain, oxidative phosphorylation, and complex I, were significantly depleted in vehicle-treated HFpEF mice compared to controls (Figure 9 and Figure 10). These mitochondrial gene sets were significantly enriched in HFpEF mice treated with TYA-11631 (Figure 9, Figure 10, and Table 5). [Table 5]

[0352] Pearson correlation coefficient analysis was performed between the expression of genes identified by GSEA analysis and different parameters of LV structure and cardiac diastolic function, including end-diastolic left ventricular posterior wall thickness (LVPWd), and isovolumic relaxation time (IVRT), or mitral valve E / e' (MV E / e'). The expression levels of several genes related to fibrosis (Col1a2, Col3a1, Fbn1, Postn, Cilp) were significantly increased in vehicle-treated HFpEF animals with diastolic dysfunction, as indicated by increased LVPWd and IVRT. TYA-11631 treatment led to a reduction in the expression of fibrosis-related genes in HFpEF mice with improved diastolic function (Figures 12A-12J, Tables 6, and 7). [Table 6] [Table 7]

[0353] The expression levels of genes related to different subunits of the mitochondrial respiratory electron transport chain (NADH:ubiquinone oxidoreductase subunits; Ndufa13, Ndufa13, Ndufa5, Ndufs7, Ndufa1, Ndufa8) were significantly reduced in vehicle-treated HFpEF animals with diastolic dysfunction, as indicated by increased LVPWd and MV E / e'. In response to TYA-11631 treatment, the expression levels of mitochondrial genes increased in HFpEF mice and positively correlated with improved diastolic function (Figures 11A-11F, Tables 8, and 9). [Table 8] [Table 9]

[0354] Effect of TYA-11631 on fibrosis genes in HFpEF mouse model by qPCR The fibrotic markers periostin (encoded by the Postn gene), collagen 3A1 (encoded by the Col3a1 gene), and collagen 1A1 (encoded by the Col1a1 gene) showed a trend towards increased expression in HFpEF mouse hearts as measured by qPCR. TYA-11631 treatment significantly reduced the expression of fibrotic genes to near healthy control levels (Table). [Table 10-1] [Table 10-2]

[0355] Effect of TYA-11631 on iPSC-CM metabolic status The metabolic state of human iPSC-derived CM treated with TYA-11631 (3 μM) was compared to vehicle-treated cells (DMSO). Seahorse oximetry was used to collect measurements of basal respiration and spare respiratory capacity. Although both groups had similar basal respiration rates, TYA-11631-treated human iPSC-derived CM had higher membrane potential and spare respiratory capacity, demonstrating greater ATP production, the ability of the cells to respond to energy stress, and a direct effect on cardiomyocytes (Figures 11G and 11H, and Table 10). [Table 11]

[0356] Thus, TYA-11631 treatment was shown to reduce activated pathogenic transcriptional signatures in mouse models of HFpEF (gene sets included hypertrophy, fibrosis, and platelet-derived growth factor receptor signaling). Targeted gene expression analysis using qPCR confirmed reduced expression of fibrotic genes that correlated with improved diastolic function. Also, based on RNA-Seq data, TYA-11631 enriched gene sets related to mitochondrial energy production in HFpEF mice. Increased expression of mitochondrial genes correlated with improved HFpEF phenotype. To functionally assess the metabolic effects of TYA-11631 in vitro, iPSC-derived CMs were used. Data indicate increased spare respiratory capacity in TYA-11631-treated human iPSC-derived CMs and improved ATP production in response to metabolic demand.

[0357] Taken together, these results show that HDAC6 selective inhibitors reverse existing diastolic dysfunction through multiple cardiac pathways related to fibrosis and mitochondrial dysfunction, both of which contribute to the pathogenesis of HFpEF.These results also confirm that HDAC6 selective inhibitors have direct cardiac benefits in HFpEF models, and that the improvements seen are due to multiple cardiac mechanisms and do not occur solely as a result of improving systemic metabolism and inflammation.

[0358] Example 6: Efficacy of HDAC6 selective inhibitors and empagliflozin in HFpEF mouse model High-fat diet and N ωThis study was designed to compare the efficacy between TYA-11018 (a selective HDAC6 inhibitor) and empagliflozin (a selective SGLT2 inhibitor, MedChemExpress LLC) in a mouse model of heart failure with preserved ejection fraction (HFpEF) induced by combined inhibition of constitutive nitric oxide synthase using -nitro l-arginine methyl ester (L-NAME). After the HFpEF phenotype was established, mice were orally administered 15 mg / kg TYA-11018 or 10 mg / kg empagliflozin, or vehicle once daily for 9 weeks. Health monitoring was performed weekly and body weights were recorded. Glucose tolerance tests were performed after the first dose. Cardiac function was recorded by echocardiography after 9 weeks of treatment. End-diastolic pressure (EDP) was measured by distal PV loops after 9 weeks of treatment.

[0359] Testing Systems Animals: A cohort of mice with an established HFpEF phenotype and age / sex-matched wild-type controls were used for the dosing study. Preparation of the HFpEF model was accomplished as previously described (Schiattarella, G et al., Nature 568, 351-356 (2019)). Briefly, 10-week-old male C57BL / 6J (catalog no. 000664) mice were purchased from Jackson Laboratory (Bar Harbor, ME). After 1 week of acclimation, one group of mice was fed a high-fat diet containing 0.07% L-NAME (LabDiet 5G5V, St Louis, MO) for 15 weeks. A control group of mice was fed a normal chow diet (LabDiet 5053, St Louis, MO). Echocardiography was performed to document HFpEF development prior to enrollment of animals in the efficacy study. Animals were housed in an animal research facility in accordance with the National Research Council of the National Academies guidelines for the care and use of laboratory animals.

[0360] HFpEF model induction Ten-week-old male C57BL / 6J (catalog no. 000664) mice were purchased from Jackson Laboratory (Bar Harbor, ME). After 1 week of acclimation, one group of mice was placed in 0.07% N ω The mice were fed a high-fat diet containing l-nitro-arginine methyl ester (L-NAME) for 15 weeks (modified from reference by Schiatarella, G. et al., Nature 568, 351-356 (2019)). A second group of mice was fed a standard chow diet as a control.

[0361] Conventional echocardiography and Doppler imaging Transthoracic echocardiography was performed using a VisualSonics Vevo2100 system with an MS400 transducer (Visual Sonics, Toronto, ON). LVEF and other indices of systolic function were obtained from short-axis M-mode scans at the mid-ventricle, as indicated by the presence of papillary muscles, in conscious, gently restrained mice. Apical four-chamber views were obtained in anesthetized mice for diastolic function measurements using pulsed wave and tissue Doppler imaging at the level of the mitral valve. Anesthesia was induced with 5% isoflurane and confirmed by the lack of response to firm pressure on one of the hind limbs. During the acquisition of echocardiograms under temperature-controlled conditions, isoflurane was reduced to 1.0–1.5% and adjusted to maintain a heart rate in the range of 450–500 beats / min. Parameters collected included left ventricular ejection fraction (LVEF), left ventricular mass (LV weight), end-diastolic left ventricular posterior wall thickness (LVPWd), peak Doppler blood inflow velocity across the mitral valve during early diastole (E wave), peak Doppler blood inflow velocity across the mitral valve during late diastole (e'), and isovolumic relaxation time (IVRT). At the end of the procedure, all mice recovered from anesthesia without difficulty.

[0362] Intraperitoneal glucose tolerance test (IP-GTT) Intraperitoneal glucose tolerance tests were performed by injection of glucose (2 g / kg in saline) after a 6-h fast (7:00 AM–1:00 PM). Tail blood glucose levels (mg / dL) were measured at 0 (baseline, before glucose administration), 15, 30, 45, 60, and 120 min after glucose administration by AimStrip Plus blood glucose strips used in the AimStrip Plus blood glucose monitoring system (Germaine Laboratories, San Antonio, TX).

[0363] Animal Enrollment and Randomization of Efficacy Studies After the HFpEF phenotype was established, using balanced echocardiographic parameters, animals on HFD / L-NAME or control diet were randomized into two groups to receive 30 mg / kg TYA-11631 or vehicle orally once daily for 9 weeks, respectively. TYA-11631 was formulated in the vehicle 5% DMSO + 45% PEG-300 + 50% purified water. Body weight and food consumption were monitored daily throughout the study.

[0364] pressure volume analysis The trachea was exposed by a midline incision from the base of the throat to just above the clavicle. Mice were intubated with a single piece of polyethylene-90 tubing. After the tube was secured in place by using a 6-0 silk suture, 100% oxygen was gently delivered across the opening. Ventilated mice were placed on a warming (37°C) pad. The right carotid artery was then isolated. Care was taken to prevent damage to the vagus nerve. Mice were lightly anesthetized with isoflurane and the heart rate was maintained at 450-550 beats / min. A 1.2F pressure-volume catheter (FTE-1212B-4518, Transonic, Inc, Ithaca, NY) was inserted into the right carotid artery and then advanced into the left ventricle. After the transducer was advanced into the ventricular cavity, as evidenced by a change in the pressure curve, it was firmly secured in place. Hemodynamic parameters were then recorded in a closed-chest mode.

[0365] Animal Randomization Mice with an established HFpEF phenotype were randomized to receive 15 mg / kg TYA-11018 or 10 mg / kg empagliflozin or vehicle (n=12 per group) orally once daily for 9 weeks. In the control (non-HFpEF) group, 9 mice received vehicle. [Table 12]

[0366] result A single dose of TYA-11018 improves fasting glucose and glucose tolerance to levels similar to empagliflozin A glucose tolerance test (IP-GTT) was performed 6 hours after the first dose. Animals were fasted during the 6 hours. A single oral dose of 15 mg / kg TYA-11018 significantly reduced fasting glucose and improved glucose tolerance to levels similar to empagliflozin in an established mouse HFpEF model ( FIG. 14A ).

[0367] Nine weeks of TYA-11018 treatment demonstrates comparable efficacy compared to empagliflozin in HFpEF models Echocardiography was used to measure LV structure and function after 9 weeks of treatment. M-mode echocardiography showed that TYA-11018 treatment preserved ejection fraction and significantly reduced left ventricular mass, similar to empagliflozin. Noninvasive Doppler imaging and terminal invasive catheterization analysis revealed that 9 weeks of treatment with both compounds reduced E / A, E / e' ratios, and reduced end-diastolic pressure, indicating improved LV relaxation and filling pressures. Notably, a superior reduction in E / e' was observed with TYA-11018 compared to empagliflozin (Figures 14B-14F).

[0368] TYA-11018 inhibits the upregulation of marker genes of cardiac stress and fibrosis in cardiac tissue After 9 weeks of treatment, cardiac tissue was harvested and processed for gene expression analysis. Total RNA was extracted from mouse cardiac tissue using TRIzol reagent (Invitrogen, Waltham, MA). A total of 500ng of RNA was used for reverse transcription using the SuperScript™ III First-Strand Synthesis System (Invitrogen, Waltham, MA). Real-time PCR was performed in duplicate using TaqMan Gene Expression Assay probes with specific primers for Nppb and Col3a1 gene sequences. The amount of target mRNA in the samples was estimated using the 2-ΔΔCt relative quantification method using Gapdh for normalization, and fold ratios were calculated compared to the mRNA expression levels from control samples.

[0369] TYA-11018 significantly inhibited the upregulation of Nppb and Col3a1, two marker genes commonly used to reveal cardiac stress and fibrosis, respectively, in cardiac tissue. The changes in gene expression in cardiac tissue were consistent with the improvement of cardiac structure and function. Notably, this selective HDAC6 inhibitor demonstrated superior efficacy in inhibiting these genes compared to empagliflozin. The dramatic effect of the selective HDAC6 inhibitor on the profibrotic gene Col3a1 also suggests a potential mechanism underlying the effect of HDAC6 inhibition that is distinct from SGLT2 inhibition.

[0370] In summary, the comparable efficacy observed in HFpEF models with HDAC6 inhibition and SGLT2 inhibition provides evidence for the translatability of these findings to clinical development.

[0371] Example 7: Biochemical activity and potency of various HDAC6 inhibitors of formula (I) The compounds disclosed herein, particularly compounds of formula (I), were synthesized according to the methods disclosed in PCT / US2020 / 066439, published as WO2021 / 127643(A1), which is incorporated herein by reference in its entirety. The compounds were tested in biochemical assays for potency against HDAC6 and selectivity against HDAC1. A biochemical assay using a luminescent HDAC-Glo I / II assay (Promega) was employed to measure the relative activity of HDAC6 and HDAC1 recombinant proteins. Compounds were first incubated separately in the presence of HDAC6 or HDAC1, followed by the addition of a luminescent substrate. A plate reader was used to acquire data and the biochemical IC values ​​were calculated accordingly. 50 was calculated from the data. The data is summarized in Table 13. From these studies, it was determined that the compounds of the present disclosure are selective inhibitors of HDAC6 over HDAC1, resulting in a selectivity ratio of about 5 to about 30,0000. [Table 13-1] [Table 13-2] [Table 13-3] [Table 13-4] [Table 13-5] [Table 13-6] [Table 13-7] [Table 13-8] [Table 13-9] [Table 13-10]

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[0372] Example 8: Biochemical activity and potency of various HDAC6 inhibitors of formula (II) The compounds disclosed herein, particularly compounds of formula (II), were synthesized according to the methods disclosed in WO2021 / 067859, the entirety of which is incorporated herein by reference. These compounds were tested in biochemical assays for potency against HDAC6 and selectivity against HDAC1. A biochemical assay using the luminescent HDAC-Glo I / II assay (Promega) was employed to measure the relative activity of HDAC6 and HDAC1 recombinant proteins. Compounds were first incubated separately in the presence of HDAC6 or HDAC1, followed by the addition of a luminescent substrate. A plate reader was used to acquire data and the biochemical IC values ​​were calculated accordingly. 50 was calculated from the data. The data is summarized in Table 14. From these studies, it was determined that the compounds of the present disclosure are selective inhibitors of HDAC6 over HDAC1, resulting in a selectivity ratio of about 5 to about 30,0000. [Table 14-1] [Table 14-2]

[0373] The structures, chemical names, and additional biochemical properties of the compounds described in this example are provided below. [Table 23] [Table 24-1] [Table 24-2] [Table 24-3] [Table 24-4] [Table 25-1] [Table 25-2]

Table 25-3

Table 26

[0374] References Cao, D. J., Wang, Z. V., Battiprolu, P. K., Jiang, N., Morales, C. R., Kong, Y., Rothermel, B. A., Gillette, T. G., Hill, J. A., 2011. Histone deacetylase (HDAC) inhibitors attenuate cardiac hypertrophy by suppressing autophagy. Proc. Natl. Acad. Sci. 108, 4123 - 4128. https: / / doi.org / 10.1073 / pnas.1015081108 Demos - Davies, K. M., Ferguson, B. S., Cavasin, M. A., Mahaffey, J. H., Williams, S. M., Spiltoir, J. I., Schuetze, K. B., Horn, T. R., Chen, B., Ferrara, C., Scellini, B., Piroddi, N., Tesi, C., Poggesi, C., Jeong, M. Y., McKinsey, T. A., 2014. HDAC6 contributes to pathological responses of heart and skeletal muscle to chronic angiotensin - II signaling. Am. J. Physiol. - Heart Circ. Physiol. 307, H252 - H258. https: / / doi.org / 10.1152 / ajpheart.00149.2014 Haberland,M.,Montgomery,R.L.,Olson,E.N.,2009.The many roles of histone deacetylases in development and physiology:implications for disease and therapy.Nat.Rev.Genet.10,32-42.https: / / doi.org / 10.1038 / nrg2485 Hubbert,C.,Guardiola,A.,Shao,R.,Kawaguchi,Y.,Ito,A.,Nixon,A.,Yoshida,M.,Wang,X.-F.,Yao,T.-P.,2002.HDAC6 is a microtubule-associated deacetylase.Nature417,455-458.https: / / doi.org / 10.1038 / 417455a Jeong,M.Y.,Lin,Y.H.,Wennersten,S.A.,Demos-Davies,K.M.,Cavasin,M.A.,Mahaffey,J.H.,Monzani,V.,Saripalli,C.,Mascagni,P.,Reece,T.B.,Ambardekar,A.V.,Granzier,H.L.,Dinarello,C.A.,McKinsey,T.A.,2018.Histone deacetylase activity governs diastolic dysfunction through a nongenomic mechanism.Sci.Transl.Med.10,eaao0144.https: / / doi.org / 10.1126 / scitranslmed.aao0144 McLendon,P.M.,Ferguson,B.S.,Osinska,H.,Bhuiyan,M.S.,James,J.,McKinsey,T.A.,Robbins,J.,2014.Tubulin hyperacetylation is adaptive in cardiac proteotoxicity by promoting autophagy.Proc.Natl.Acad.Sci.111,E5178-E5186.https: / / doi.org / 10.1073 / pnas.1415589111 Nagata,S.,Marunouchi,T.,Tanonaka,K.,2019.Histone Deacetylase Inhibitor SAHA Treatment Prevents the Development of Heart Failure after Myocardial Infarction via an Induction of Heat-Shock Proteins in Rats.Biol.Pharm.Bull.42,453-461.https: / / doi.org / 10.1248 / bpb.b18-00785 Zhang,Y.,Kwon,S.,Yamaguchi,T.,Cubizolles,F.,Rousseaux,S.,Kneissel,M.,Cao,C.,Li,N.,Cheng,H.-L.,Chua,K.,Lombard,D.,Mizeracki,A.,Matthias,G.,Alt,F.W.,Khochbin,S.,Matthias,P.,2008.Mice Lacking Histone Deacetylase 6 Have Hyperacetylated Tubulin but Are Viable and Develop Normally.Mol.Cell.Biol.28,1688-1701.https: / / doi.org / 10.1128 / MCB.01154-06 Yoon S,Eom GH.HDAC and HDAC Inhibitor:From Cancer to Cardiovascular Diseases.Chonnam Med J.2016 Jan;52(1):1-11.https: / / doi.org / 10.4068 / cmj.2016.52.1.1 Schiattarella,G.G.,Tong,D.,Hill,J.A.,2020.Can HFpEF and HFrEF Coexist? 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[0375] Incorporation by reference All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entirety for all purposes. However, mention of any references, articles, publications, patents, patent publications, and patent applications cited herein is not and should not be construed as an admission or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country throughout the world.

Claims

1. 1. A pharmaceutical composition for use in the treatment or prevention of heart failure with preserved ejection fraction (HFpEF) in a subject in need thereof, the pharmaceutical composition comprising an HDAC6 inhibitor, The HDAC inhibitor is a compound according to formula I(y): 【Chemistry 144】 or a pharma- ceutically acceptable salt thereof; During the ceremony, X 1 is S; R a is selected from the group consisting of H, halogen, and C 1-3 alkyl; R 1 is 【Chemistry 145】 and R 2 is selected from the group consisting of C 1-5 alkyl, C 1-5 alkoxy, and C 3-12 cycloalkyl, each of which is optionally substituted with one or more of oxo, halogen, C 1-5 alkyl, C 1-5 haloalkyl, C 1-5 alkoxy, C 1-5 haloalkoxy, C 3-12 cycloalkyl, cyano, and amido; R 3 is H or C 1-5 alkyl; R 4 is selected from the group consisting of C 1-5 alkyl, —(SO 2 )R 2 , —(SO 2 )NR 2 R 3 , and —(CO)R 2 ; R 5 is C 3-12 cycloalkyl, C 6-12 aryl, or C 5-14 heteroaryl having 1 or 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more of oxo, halogen, C 1-5 alkyl, C 1-5 haloalkyl, C 1-5 alkoxy, C 1-5 haloalkoxy, C 3-12 cycloalkyl, 3- to 12-membered heterocyclyl having 1 or 2 heteroatoms independently selected from N, O, and S, 5- to 14-membered heteroaryl having 1 or 2 heteroatoms independently selected from N, O, and S, ester, amido, cyano, C 1-5 cyanoalkyl, and C 1-5 alkylamino; A pharmaceutical composition, wherein R 4 and R 5 together with the atom to which they are attached form a 3-12 membered heterocyclyl having 1 or 2 heteroatoms independently selected from N, O and S, each of which is optionally substituted with one or more of oxo, halogen, C 1-5 alkyl, C 1-5 haloalkyl, C 1-5 alkoxy, C 1-5 haloalkoxy, cyano, C 3-12 cycloalkyl, 3-12 membered heterocyclyl having 1 or 2 heteroatoms independently selected from N, O and S, and 5-14 membered heteroaryl having 1 or 2 heteroatoms independently selected from N, O and S.

2. The pharmaceutical composition of claim 1, wherein R a is H.

3. R 1 is 【Chemistry 159】 The pharmaceutical composition of claim 1 , 4. The pharmaceutical composition of claim 1, wherein R 4 is --(SO 2 )R 2 .

5. The pharmaceutical composition of claim 4, wherein -(SO 2 )R 2 is -(SO 2 )(C 1-5 alkyl), -(SO 2 )(C 1-3 alkylene 3- to 12-membered heterocyclyl having 1 or 2 heteroatoms independently selected from N, O and S), -(SO 2 )(C 1-5 haloalkyl), -(SO 2 )(C 1-5 haloalkoxy), or -(SO 2 )(C 3-12 cycloalkyl).

6. R 4 is -(CO)CH 3 , -(CO)CH 2 CH 3 , -(CO)CH 2 CH 2 CH 3 , -(SO 2 )CH 3 , -(SO 2 )CH 2 CH 3 , -(SO 2 )CH 2 CH 2 CH 3 , and 【Chemistry 160】 The pharmaceutical composition of claim 5, selected from the group consisting of:

7. The pharmaceutical composition of claim 1, wherein R 5 is a 5-6 membered heteroaryl having 1 or 2 heteroatoms selected from nitrogen, oxygen, and sulfur.

8. The pharmaceutical composition of claim 7, wherein the heteroaryl is a 5- to 6-membered heteroaryl having 1 or 2 nitrogen atoms.

9. The 5- to 6-membered heteroaryl is 【Chemistry 161】 9. The pharmaceutical composition of claim 8, wherein R b is halogen, C 1-5 alkyl, C 1-5 alkoxy, C 3-12 cycloalkyl, -CN, C 1-5 haloalkyl, or C 1-5 haloalkoxy; and m is 0 or 1.

10. The pharmaceutical composition of claim 9, wherein R b is F, Cl, -CH 3 , -CH 2 CH 3 , -CF 3 , -CHF 2 , -CF 2 CH 3 , -CN, -OCH 3 , -OCH 2 CH 3 , -OCH(CH 3 ) 2 , -OCF 3 , -OCHF 2 , -OCH 2 CF 2 H, and cyclopropyl.

11. The pharmaceutical composition of claim 1, wherein R 5 is C 6-12 aryl selected from the group consisting of phenyl, 3-chlorophenyl, 3-chloro-4-fluorophenyl, 3-trifluoromethylphenyl, 3,4-difluorophenyl, and 2,6-difluorophenyl.

12. The HDAC6 inhibitor, 【Table 30-1】 【Table 30-2】 【Table 30-3】 【Table 30-4】 【Table 30-5】 【Table 30-6】 【Table 30-7】 【Table 30-8】 【Table 30-9】 【Table 30-10】 【Table 30-11】 【Table 30-12】 【Table 30-13】 【Table 30-14】 【Table 30-15】 【Table 30-16】 【Table 30-17】 【Table 30-18】 【Table 30-19】 【Table 30-20】 【Table 30-21】 【Table 30-22】 【Table 30-23】 【Table 30-24】 【Table 30-25】 【Table 30-26】 【Table 30-27】 【Table 30-28】 【Table 30-29】 【Table 30-30】 【Table 30-31】 【Table 30-32】 【Table 30-33】 【Table 30-34】 【Table 30-35】 【Table 30-36】 【Table 30-37】 【Table 30-38】 【Table 30-39】 【Table 30-40】 【Table 30-41】 【Table 30-42】 【Table 30-43】 【Table 30-44】 【Table 30-45】 【Table 30-46】 【Table 30-47】 【Table 30-48】 【Table 30-49】 【Table 30-50】 【Table 30-51】 【Table 30-52】 【Table 30-53】 【Table 30-54】 【Table 30-55】 or a pharma- ceutically acceptable salt thereof.

13. The HDAC6 inhibitor is 【Chemistry 142】 or a pharma- ceutically acceptable salt thereof.

14. The compound is 【Chemistry 148】 or a pharma- ceutically acceptable salt thereof.

15. The compound is 【Chemistry 149】 or a pharma- ceutically acceptable salt thereof.

16. The compound is 【Chemistry 150】 or a pharma- ceutically acceptable salt thereof.

17. The compound is 【Chemistry 151】 or a pharma- ceutically acceptable salt thereof.

18. The compound is 【Chemistry 152】 or a pharma- ceutically acceptable salt thereof.

19. The compound is 【Chemistry 153】 or a pharma- ceutically acceptable salt thereof.

20. The compound is 【Chemistry 154】 or a pharma- ceutically acceptable salt thereof.

21. The compound is 【Chemistry 155】 or a pharma- ceutically acceptable salt thereof.

22. The compound is 【Chemistry 156】 or a pharma- ceutically acceptable salt thereof.

23. The compound is 【Chemistry 157】 or a pharma- ceutically acceptable salt thereof.

24. A pharmaceutical composition described in any one of claims 1 to 23, wherein the subject has hypertension or is at risk of hypertension.

25. A pharmaceutical composition described in any one of claims 1 to 23, wherein the subject has diabetes mellitus or is at risk of diabetes mellitus.

26. The pharmaceutical composition described in any one of claims 1 to 23, wherein the subject has coronary artery disease (CAD) or is at risk for coronary artery disease (CAD).

27. ​​A pharmaceutical composition described in any one of claims 1 to 23, wherein the subject has or is at risk of valvular heart disease.

28. A pharmaceutical composition described in any one of claims 1 to 23, wherein the subject has atrial fibrillation or is at risk of atrial fibrillation.

29. The pharmaceutical composition of any one of claims 1 to 23, wherein the use comprises treating or preventing at least one symptom of HFpEF.

30. The pharmaceutical composition of any one of claims 1 to 23, wherein the use comprises reducing left ventricular (LV) mass.

31. The pharmaceutical composition of any one of claims 1 to 23, wherein the use comprises reducing LV wall thickness.

32. The pharmaceutical composition of any one of claims 1 to 23, wherein the use comprises improving LV relaxation.

33. The pharmaceutical composition of any one of claims 1 to 23, wherein the use comprises improving LV filling pressure.

34. A pharmaceutical composition described in any one of claims 1 to 23, wherein the use comprises preventing heart failure in the subject.

35. The pharmaceutical composition described in any one of claims 1 to 23, wherein the subject is a human.

36. The pharmaceutical composition of claim 34, wherein the subject is at least 65 or 70 years of age.

37. A pharmaceutical composition for use in treating or preventing a metabolic disease in a subject in need thereof, the pharmaceutical composition comprising an HDAC6 inhibitor, The HDAC inhibitor is a compound according to formula I(y): 【Chemistry 144】 or a pharma- ceutically acceptable salt thereof; During the ceremony, X 1 is S; R a is selected from the group consisting of H, halogen, and C 1-3 alkyl; R 1 is 【Chemistry 145】 and R 2 is selected from the group consisting of C 1-5 alkyl, C 1-5 alkoxy, and C 3-12 cycloalkyl, each of which is optionally substituted with one or more of oxo, halogen, C 1-5 alkyl, C 1-5 haloalkyl, C 1-5 alkoxy, C 1-5 haloalkoxy, C 3-12 cycloalkyl, cyano, and amido; R 3 is H or C 1-5 alkyl; R 4 is selected from the group consisting of C 1-5 alkyl, —(SO 2 )R 2 , —(SO 2 )NR 2 R 3 , and —(CO)R 2 ; R 5 is C 3-12 cycloalkyl, C 6-12 aryl, or C 5-14 heteroaryl having 1 or 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more of oxo, halogen, C 1-5 alkyl, C 1-5 haloalkyl, C 1-5 alkoxy, C 1-5 haloalkoxy, C 3-12 cycloalkyl, 3- to 12-membered heterocyclyl having 1 or 2 heteroatoms independently selected from N, O, and S, 5- to 14-membered heteroaryl having 1 or 2 heteroatoms independently selected from N, O, and S, ester, amido, cyano, C 1-5 cyanoalkyl, and C 1-5 alkylamino; A pharmaceutical composition, wherein R 4 and R 5 together with the atom to which they are attached form a 3-12 membered heterocyclyl having 1 or 2 heteroatoms independently selected from N, O and S, each of which is optionally substituted with one or more of oxo, halogen, C 1-5 alkyl, C 1-5 haloalkyl, C 1-5 alkoxy, C 1-5 haloalkoxy, cyano, C 3-12 cycloalkyl, 3-12 membered heterocyclyl having 1 or 2 heteroatoms independently selected from N, O and S, and 5-14 membered heteroaryl having 1 or 2 heteroatoms independently selected from N, O and S.

38. The pharmaceutical composition of claim 37, wherein the metabolic disease is a metabolic disease associated with obesity, optionally diet-induced obesity.

39. The pharmaceutical composition described in claim 37, wherein the metabolic disease is diabetes, prediabetes, diabetic cardiomyopathy, metabolic syndrome, hypertension, hypertriglyceridemia, or dyslipidemia.

40. The pharmaceutical composition described in claim 39, wherein the metabolic disease is diabetes.

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