HDAC6 inhibitors for use in the treatment of dilated cardiomyopathy - Patent Application 20070123333

Abstract

Provided herein is a method for treating or preventing dilated cardiomyopathy (DCM) using HDAC6 inhibitors.Various HDAC6 inhibitors are described for use in treating or preventing DCM.In one aspect, described herein is a method for treating a human patient by orally administering an HDAC6 inhibitor, such as an inhibitor of formula (I) or formula (II).In one aspect, described herein is a method for treating a human patient with DCM associated with reduced ejection fraction.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 178,901, filed April 23, 2021, which is incorporated herein by reference in its entirety.

[0002] The present disclosure relates to the treatment of dilated cardiomyopathy (DCM). [Background technology]

[0003] Dilated cardiomyopathy (DCM) is a form of myocardial weakness characterized by reduced cardiac output and thinning and enlargement of the left ventricular cavity (McNally et al., 2013; Villard et al., 2011). DCM affects approximately 1 in 2500 adults (Villard et al., 2011), accounts for 30%-40% of all heart failure cases in clinical trials, and is the leading cause of heart transplantation (Everly, 2008; Haas et al., 2015).

[0004] Current treatments for heart failure include angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, beta-blockers, aldosterone antagonists, vasodilators, angiotensin receptor-neprilysin inhibitors, and sodium-glucose cotransporter 2 inhibitors. These treatments primarily relieve symptoms and do not target the underlying molecular mechanisms associated with genetic forms of heart failure (Cleland et al., 2020).

[0005] Approximately one-third of DCM patients have a genetic cause. Familial DCM accounts for 30%-50% of all DCM cases and has an autosomal dominant inheritance pattern (Haas et al., 2015). DCM genotypes have been associated with more than 50 genes, and more than 50% of patients with DCM have at least one mutation in one of these genes (Haas et al., 2015; McNally et al., 2013). Several of these DCM-associated genes encode central regulators of protein quality control, and mutations in these genes lead to protein aggregation and accumulation of misfolded proteins (Fang et al., 2017; Sturner and Behl, 2017).

[0006] There is an unmet need for treatment of DCM. Summary of the Invention [Means for solving the problem]

[0007] The present disclosure relates generally to methods of treating dilated cardiomyopathy by administering an HDAC6 inhibitor, such as TYA-018 or an analog thereof.

[0008] In one aspect, the disclosure provides a method of treating or preventing dilated cardiomyopathy in a subject in need thereof, the method comprising administering a therapeutically effective amount of an HDAC6 inhibitor.

[0009] In some embodiments, the present disclosure provides a method of treating or preventing dilated cardiomyopathy associated with reduced ejection fraction in a subject in need thereof, the method comprising administering a therapeutically effective amount of an HDAC6 inhibitor.

[0010] In some embodiments, the present disclosure provides a method for treating dilated cardiomyopathy in a subject in need thereof, the method comprises administering a 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 general formula: [ka]

[0011] In some embodiments, the present disclosure provides a method for preventing dilated cardiomyopathy in a subject in need thereof, the method comprises administering a 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 general formula: [ka]

[0012] In some embodiments, the HDAC6 inhibitor is a compound according to formula (I): [ka] R 1 is selected from the group consisting of: [Table 11] 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 3together 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, together with the atoms to which they are attached, they form a cycloalkyl or heterocyclyl, Y is CR 2 or N, n is selected from 0, 1, and 2.

[0013] 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.

[0014] In some embodiments of Formula (Ik), R b is H, halogen, haloalkyl, or haloalkoxy.

[0015] In some embodiments of Formula (Ik), R 4 is optionally substituted alkyl or cycloalkyl.

[0016] 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.

[0017] In some embodiments of formula (Ik-1), R b is H, halogen, haloalkyl, or haloalkoxy.

[0018] In some embodiments of formula (Ik-1), R 4 is optionally substituted alkyl or cycloalkyl.

[0019] In some embodiments of formula (Ik-1), R 4is alkyl.

[0020] 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.

[0021] In some embodiments of formula (Ik-2), R b is H, halogen, haloalkyl, or haloalkoxy.

[0022] In some embodiments of formula (Ik-2), R 4 is optionally substituted alkyl.

[0023] 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 taken together with the atom to which they are attached form a heterocyclyl, each of which is optionally substituted.

[0024] In some embodiments of formula I(y), R a is H.

[0025] In some embodiments of formula I(y), R 1 teeth [ka] It is.

[0026] In some embodiments of formula I(y), R 4 Ha-(SO2)R 2 It is.

[0027] In some embodiments of Formula I(y), -(SO2)R 2 is -(SO2)alkyl, -(SO2)alkyleneheterocyclyl, -(SO2)haloalkyl, -(SO2)haloalkoxy, or -(SO2)cycloalkyl.

[0028] In some embodiments of formula I(y), R 5 is heteroaryl.

[0029] In some embodiments of Formula I(y), the heteroaryl is a 5-6 membered heteroaryl.

[0030] 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.

[0031] 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.

[0032] 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.

[0033] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0034] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0035] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0036] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0037] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0038] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0039] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0040] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0041] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0042] In some embodiments of formula I(y), the compound is [ka] or a pharma- ceutically acceptable salt thereof.

[0043] In some embodiments, the HDAC6 inhibitor is selected from the group consisting of: [Table 6-1] [Table 6-2]

[0044] In some embodiments, the HDAC6 inhibitor is [ka] (TYA-018) or its analogues.

[0045] In some embodiments, the HDAC6 inhibitor is TYA-018.

[0046] In some embodiments, the HDAC6 inhibitor is a compound according to formula (II): [ka] 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; R 1 and R 2together 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; 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.

[0047] 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.

[0048] In some embodiments, the HDAC6 inhibitor is tubastatin A.

[0049] In some embodiments, the HDAC6 inhibitor is ricorinostat.

[0050] In some embodiments, the HDAC6 inhibitor is CAY10603.

[0051] In some embodiments, the HDAC6 inhibitor is nextulastat A.

[0052] In some embodiments, the HDAC6 inhibitor is at least 100-fold selective for HDAC6 over all other isozymes of HDAC.

[0053] 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.

[0054] In one aspect, the disclosure provides a method of treating or preventing dilated cardiomyopathy in a subject in need thereof, the method comprising administering a gene silencing agent, such as an RNA silencing agent (e.g., siRNA).

[0055] In some embodiments, the dilated cardiomyopathy is familial dilated hypertrophic cardiomyopathy.

[0056] In some embodiments, the dilated cardiomyopathy is dilated cardiomyopathy caused by one or more BLC2-associated athanogene3 (BAG3) mutations.

[0057] In some embodiments, the subject has a deleterious mutation in the BAG3 gene. E455K Has a mutation.

[0058] In some embodiments, the dilated cardiomyopathy is dilated cardiomyopathy caused by one or more muscle LIM protein (MLP) mutations.

[0059] In some embodiments, the subject has a deleterious mutation in the CSPR3 gene, which encodes MLP.

[0060] In some embodiments, the subject is a human.

[0061] In some embodiments, administration to a subject is oral.

[0062] In some embodiments, the methods restore the subject's ejection fraction to at least that of a subject without dilated cardiomyopathy.

[0063] In some embodiments, the methods increase the subject's ejection fraction compared to the subject's ejection fraction before treatment.

[0064] In some embodiments, the methods restore the subject's ejection fraction to at least about 20%, at least about 20%, at least about 30%, at least about 40%, or at least about 50%.

[0065] In some embodiments, the method increases the subject's ejection fraction by at least about 5%, at least about 10%, at least about 20%, at least about 30%, or at least about 40%.

[0066] In some embodiments, the method reduces HDAC6 activity in the subject's heart. In some embodiments, the method substantially eliminates HDAC6 activity in the subject's heart.

[0067] In some embodiments, the method prevents heart failure in a subject.

[0068] In some embodiments, the method reduces left ventricular diameter at diastolic pressure (LVIDd) in a subject.

[0069] In some embodiments, the methods reduce left ventricular systolic dimensions (LVIDs) in a subject.

[0070] In some embodiments, the method reduces left ventricular mass in the subject.

[0071] 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.

[0072] In another aspect, the disclosure provides an HDAC6 inhibitor for use in a method for treating dilated cardiomyopathy.

[0073] In another aspect, the disclosure provides a pharmaceutical composition for use in a method for treating dilated cardiomyopathy, comprising an HDAC6 inhibitor.

[0074] In another aspect, the disclosure provides a kit comprising an HDAC6 inhibitor and instructions for use in a method for treating dilated cardiomyopathy.

[0075] In another aspect, the disclosure provides the use of an HDAC6 inhibitor in the treatment of dilated cardiomyopathy.

[0076] In another aspect, the disclosure provides a method for identifying a compound for the treatment of dilated cardiomyopathy, the method comprising contacting a cell culture comprising a cell having an inactivating mutation in BAG3 with each member of a plurality of candidate compounds and selecting a compound that reduces sarcomere damage in the cell.

[0077] In another aspect, the disclosure provides a method of treating dilated cardiomyopathy in a subject in need thereof, the method comprising: (a) contacting a cell culture comprising cells having an inactivating mutation in BAG3 with each member of a plurality of candidate compounds and selecting a compound that reduces sarcomere damage; and (b) administering a therapeutically effective amount of the selected compound to the subject. [Brief description of the drawings]

[0078] [Figure 1] 1 shows a pictorial summary of the experimental methods used herein. [Figure 2A]Protein quantification using immunostaining of iPSC-CMs treated with SCR and BAG3 siRNA is shown. BAG3 protein levels were reduced by approximately 75%. In addition, MYBPC3 and p62 protein levels were reduced, suggesting defects in sarcomeric and autophagic flux. Error bars = SD. ****P<0.0001. [Figure 2B] 1 shows a pictorial summary of the experimental methods used herein. [Figure 2C] Quantification of sarcomere damage in iPSC-CMs treated with SCR or BAG3 siRNA. The number of damaged iPSC-CMs increased as a function of time in BAG3 knockdown (KD) cells. Error bars = SD. ****P<0.0001. [Figure 3A] A schematic diagram of the high content screening method using iPSC-CMs is shown. [Figure 3B] We show that an unbiased screen was performed using a library of 5500 bioactive compounds. iPSC-CMs were treated with BAG3 siRNA and compounds at a concentration of 1 μM. Hits were first identified using deep learning on control iPSC-CMs treated with either SCR or BAG3 siRNA. The hit threshold was set at a cardiomyocyte score of 0.3. [Figure 3C] The top 24 compounds are histone deacetylase (HDAC) and microtubule inhibitors. In addition, three known heart failure medications were identified: sotalol (a beta-blocker and K channel blocker), omecamtiv mecarbil (a cardiac myosin activator), and anagrelide (a PDE3 inhibitor). [Figure 4A] iPSC-CMs were treated with a panel of pan- and isozyme-specific HDAC inhibitors and protein levels were quantified 4 days after treatment using immunochemistry at 5 doses ranging from 10 nM to 1000 nM. BAG3 protein levels were not increased in cells treated with HDAC inhibitors. Bortezomib (known to increase BAG3 expression) was used as a positive control. [Figure 4B]The same panel of HDAC inhibitors was used at 100 nM concentration and RNA expression was quantified using qPCR 2 days after drug treatment. HDAC inhibitors did not activate BAG3 expression at the transcriptional level. Bortezomib (known to increase BAG3 expression) was used as a positive control. Error bars = SD. [Figure 5A] (Figures 5A-5C) Target validation studies show that inhibition of HDAC6 is sufficient to protect against sarcomere damage in BAG3-deficient iPSC-CMs. (Figure 5A) Top compound classes from the library screen (HDAC inhibitors, microtubule inhibitors) and two cardiovascular standard of care drugs identified from the screen [omecamtib mecarbil (omecamtib) and sotalol] were validated at a dose of 1 µM using Cardiomyocyte Score. Data from 1-2 independent biological replicates. n=4-16 technical replicates per condition. Error bars = SD. [Figure 5B] Further validation using siRNA showed that knockdown of HDAC6 provided protection from sarcomere damage in BAG3KD iPSC-CMs using a cardiomyocyte score by deep learning algorithm. Data from 2–7 independent biological replicates. n=4–16 technical replicates per condition. Error bars=SD. ****P<0.0001. [Figure 5C] Representative immunostaining of anti-MYBPC3 in iPSC-CMs treated with scrambled (SCR), BAG3, or BAG3+HDAC6 siRNA. Arrows indicate sarcomere damage. Scale bar = 50 μm. [Figure 6A](Figures 6A-6C) We show that siRNA knockdown of HDAC6 is essential and sufficient for protection from cardiomyocyte injury induced by BAG3 loss of function. (Figure 6A) Cardiomyocyte scores of iPSC-CMS treated with four siRNAs targeting HDAC1-HDAC11 (individual and pooled (p)), as well as BAG3 siRNA. Two independent siRNAs, one targeting HDAC6 and one as a pool, showed improved cardiomyocyte scores. n=4-16 technical replicates per condition. Error bars=SD. ***P<0.001. [Figure 6B] Immunocytochemistry showing that BAG3 protein levels were approximately 60% after knockdown. Knockdown of HDAC1–HDAC11 by BAG3 did not increase BAG3 levels. n = 5–16 technical replicates. Error bars = SD. ***P<0.001. [Figure 6C] Immunocytochemistry showing that knockdown of HDAC3 and HDAC6 increased tubulin acetylation (Ac-tubulin) in iPSC-CMs. n = 5–16 technical replicates. Error bars = SD. [Figure 7A] (Figures 7A-7D) TYA-018 is a highly selective HDAC6 inhibitor. (Figure 7A) HDAC6 (on-target) and HDAC1 (off-target) inhibition curves after treatment with divinostat (pan-HDAC inhibitor), tubastatin A (HDAC6-selective inhibitor), and TYA-018 (HDAC6-selective inhibitor) by biochemical assays using recombinant human HDAC6 and HDAC1 deacetylase activities. Error bars = SD. [Figure 7B] Cell-based assays in iPSC-CMs show dose-response curves of tubulin acetylation (Ac-tubulin) as a function of drug concentration. Based on EC50 calculations, divinostat, tubastatin A, and TYA-018 have similar ranges of cellular potency against HDAC6 (range 0.1 μM-0.3 μM), with TYA-018 being the most potent. Error bars = SD. EC50, half maximal effective concentration. [Figure 7C]Immunostaining of iPSC-CMs treated with 5.5 μM of each drug stained with anti-Ac-tubulin antibody. Scale bar = 200 μm. [Figure 7D] Western blot of iPSC-CMs treated with TYA-018 (HDAC6-specific inhibitor) stained with monoclonal anti-Ac-lysine. Givinostat (Giv; pan-HDAC inhibitor control) showed both on-target (Ac-tubulin staining) and off-target (Ac-histone H3 and H4 staining) activity. TYA-018 shows only specific on-target activity with no detectable off-target activity, even at 33 μM. [Figure 8A] (Figures 8A-8C) TYA-018 is a highly selective HDAC6 inhibitor, as measured by biochemical and cell-based assays. (Figure 8A) Biochemical assay measuring HDAC1-HDAC11 deacetylase activity in the presence of divinostat (a pan-HDAC inhibitor), tubastatin A (a HDAC6-selective inhibitor), and TYA-018 (a HDAC6-selective inhibitor). [Figure 8B] Selectivity for HDAC6 activity demonstrated that TYA-018 has greater than 2500-fold selectivity for HDAC6 over other HDACs. [Figure 8C] Pro-BNP in iPSC-CMs after 4 days of incubation with the drug shows that TYA-018 does not induce cell stress. Error bars = SD. [Figure 9A] (Figures 9A-9M) Givinostat and Tubastatin A protect cardiac function in BAG3cKO mice. (Figure 9A) Schematic of drug treatment in the BAG3cKO mouse model. Daily dosing began at 1 month of age. Givinostat (a pan-HDAC inhibitor) was administered daily by oral gavage (PO) at 30 mg / kg. Tubastatin A (a HDAC6-selective inhibitor) was administered daily by intraperitoneal injection (IP) at 50 mg / kg. [Figure 9B]Ejection fraction showed that daily dosing with Givinostat (Giv) protected cardiac function over the 10-week dosing period. Error bars = SEM. ***P<0.001, ****P<0.0001. [Figure 9C] Ejection fraction was monitored from the first day of dosing and delta ejection fraction was measured. Over the course of 10 weeks, cardiac function declined by 0.6% in the givinostat-treated group (not significant), while in the vehicle-treated group it declined by an average of 23.9% (****P<0.0001). Error bars=SEM. [Figure 9D] Ejection fraction (FIG. 9D) and delta ejection fraction (FIG. 9E) at 3.5 months of age and 10 weeks of treatment (compared to pre-drug baseline) indicate that givinostat provides protection from decline in cardiac function in BAG3cKO mice. Error bars=SEM. ****P<0.0001. [Figure 9E] Ejection fraction (FIG. 9D) and delta ejection fraction (FIG. 9E) at 3.5 months of age and 10 weeks of treatment (compared to pre-drug baseline) indicate that givinostat provides protection from decline in cardiac function in BAG3cKO mice. Error bars=SEM. ****P<0.0001. [Figure 9F] In BAG3cKO mice treated with givinostat, left ventricular diastolic diameter (LVIDd) (FIG. 9F) and left ventricular systolic diameter (LVIDs) (FIG. 9G) were significantly reduced, approaching the levels seen in WT littermates. Error bars = SEM. *P<0.05, ***P<0.001. [Figure 9G] In BAG3cKO mice treated with givinostat, left ventricular diastolic diameter (LVIDd) (FIG. 9F) and left ventricular systolic diameter (LVIDs) (FIG. 9G) were significantly reduced, approaching the levels seen in WT littermates. Error bars = SEM. *P<0.05, ***P<0.001. [Figure 9H] Ejection fraction showed that daily dosing of Tubastatin A (TubA) protected cardiac function over the 10-week dosing period. Error bars = SEM. *P<0.05, **P<0.01, ***P<0.001. [Figure 9I]Ejection fraction was tracked from the first day of dosing and delta ejection fraction was measured. Over a 10-week period, cardiac function was reduced by 1.7% in BAG3cKO mice treated with Tubastatin A (not significant), whereas it was reduced by 21.5% in vehicle-treated mice (**P<0.01). Error bars=SEM. [Figure 9J] Ejection fraction (FIG. 9J) and delta ejection fraction (FIG. 9K) at 3.5 months of age and 10 weeks of treatment (compared to pre-drug baseline) indicate that Tubastatin A provides protection from decline in cardiac function in BAG3cKO mice. **P<0.01, ***P<0.001. [Figure 9K] Ejection fraction (FIG. 9J) and delta ejection fraction (FIG. 9K) at 3.5 months of age and 10 weeks of treatment (compared to pre-drug baseline) indicate that Tubastatin A provides protection from decline in cardiac function in BAG3cKO mice. **P<0.01, ***P<0.001. [Figure 9L] Tubastatin A significantly reduced LVIDd (FIG. 9L) and LVIDs (FIG. 9M) in BAG3cKO mice to the levels seen in their WT littermates. Error bars=SEM. **P<0.01. [Figure 9M] Tubastatin A significantly reduced LVIDd (FIG. 9L) and LVIDs (FIG. 9M) in BAG3cKO mice to the levels seen in their WT littermates. Error bars=SEM. **P<0.01. [Figure 10A] (Figures 10A-10I) BAG3E455K mice develop heart failure and are protected by administration of Tubastatin A. (Figure 10A) Schematic of the BAG3E455K mouse model in which a LoxP site was introduced into the WT BAG3 allele, which was removed after αMHC-cre-driven excision, resulting in expression of the E455K mutant form of BAG3 (BAG3μ) in the heart. BAG3 (WT and E445K) is expressed in other tissues. [Figure 10B]Treatment with Tubastatin A (TubA; a HDAC6-selective inhibitor) was initiated at 3 months of age. Daily dosing at 50 mg / kg significantly protected mice from the decline in cardiac function compared to the vehicle-treated group. Error bars = SEM. *P<0.05. [Figure 10C] Ejection fraction was tracked from the first day of dosing and delta ejection fraction was measured. Data show that there was a 10.1% decline in mice treated with Tubastatin A over a 6-week period, compared with a 29.0% decline in the vehicle-treated group. Error bars = SEM. *P<0.05, **P<0.01. [Figure 10D] Ejection fraction and delta ejection fraction at 4.5 months of age and 6 weeks of treatment (compared to pre-drug baseline) indicate that Tubastatin A (TubA) provides protection from decline in cardiac function in BAG3cKO mice. Error bars = SEM. *P<0.05. [Figure 10E] Ejection fraction and delta ejection fraction at 4.5 months of age and 6 weeks of treatment (compared to pre-drug baseline) indicate that Tubastatin A (TubA) provides protection from decline in cardiac function in BAG3cKO mice. Error bars = SEM. *P<0.05. [Figure 10F] Tubastatin A reduced left ventricular diastolic diameter (LVIDd) (FIG. 10F) and left ventricular systolic diameter (LVIDs) (FIG. 10G) in BAG3E455K mice. [Figure 10G] Tubastatin A reduced left ventricular diastolic diameter (LVIDd) (FIG. 10F) and left ventricular systolic diameter (LVIDs) (FIG. 10G) in BAG3E455K mice. [Figure 10H] Kaplan-Meier plots show that Tubastatin A reduced mortality in BAG3E445K mice during 6 weeks of treatment (FIG. 10H), an effect that was more pronounced in male mice (FIG. 10I). [Figure 10I] Kaplan-Meier plots show that Tubastatin A reduced mortality in BAG3E445K mice during 6 weeks of treatment (FIG. 10H), an effect that was more pronounced in male mice (FIG. 10I). [Figure 11A] (FIGS. 11A-11K) Inhibition of HDAC6 by TYA-018 protects cardiac function in BAG3cKO mice. (FIG. 11A) Schematic of drug treatment in the BAG3cKO mouse model. TYA-018, a highly selective HDAC6 inhibitor, was administered daily by oral gavage at 15 mg / kg starting when mice were 2 months of age. [Figure 11B] Daily TYA-018 dosing preserved cardiac function as measured by ejection fraction over the 8-week dosing period. Error bars = SEM. *P<0.05, **P<0.01. [Figure 11C] Ejection fraction was monitored from the first day of dosing and delta ejection fraction was measured. Over the course of 8 weeks, cardiac function did not decline in the TYA-018-treated group, whereas it declined by 19.1% in the vehicle-treated group. Error bars = SEM. *P<0.05, **P<0.01. [Figure 11D] Ejection fraction (FIG. 11D) and delta ejection fraction (FIG. 11E) at 4 months of age and 8 weeks of treatment (compared to pre-drug baseline) indicate that TYA-018 provides protection from decline in cardiac function in BAG3cKO mice. Error bars=SEM. **P<0.01. [Figure 11E] Ejection fraction (FIG. 11D) and delta ejection fraction (FIG. 11E) at 4 months of age and 8 weeks of treatment (compared to pre-drug baseline) indicate that TYA-018 provides protection from decline in cardiac function in BAG3cKO mice. Error bars=SEM. **P<0.01. [Figure 11F] In BAG3cKO mice, left ventricular diastolic diameter (LVIDd) (FIG. 6F) and left ventricular systolic diameter (LVIDs) (FIG. 6G) were reduced by TYA-018, with levels approaching those seen in WT littermates. Error bars = SEM. *P<0.05. [Figure 11G] In BAG3cKO mice, left ventricular diastolic diameter (LVIDd) (FIG. 6F) and left ventricular systolic diameter (LVIDs) (FIG. 6G) were reduced by TYA-018, with levels approaching those seen in WT littermates. Error bars = SEM. *P<0.05. [Figure 11H] Hearts from all three study groups were analyzed using RNA-Seq. Principal component analysis of all coding genes showed an overall correction of BAG3cKO+TYA-018 coding genes towards WT mice. Veh, vehicle. [Figure 11I] RNA-Seq analysis shows that NPPB expression was increased approximately four-fold in BAG3cKO mice compared to WT mice at 4 months of age. TYA-018 treatment reduced NPPB levels by 2-fold in BAG3cKO mice. The levels of NPPB in BAG3cKO+TYA-018 mice were inversely correlated with cardiac function. [Figure 11J] Heatmap of RNA-Seq analysis from a selected number of genes. Data show correction of key sarcomere genes (MYH7, TNNI3, and MYL3) and genes regulating mitochondrial function and metabolism (CYC1, NDUFS8, NDUFB8, PPKARG2) in BAG3cKO+TYA-018 mice. Markers of inflammation (IL-1β, NLRP3) and apoptosis (CASP1, CAPS8) were also reduced. [Figure 11K] qPCR analysis shows an approximately 3-fold increase in NPPB expression levels in BAG3cKO mouse hearts. In BAG3cKO mice treated with TYA-018, NPPB expression levels are significantly reduced to near WT levels. Error bars = SEM. ***P<0.001. [Figure 12A] (FIGS. 12A-12B) Cardiovascular standard of care drugs do not affect Ac-tubulin and HDAC6 expression. (FIG. 12A) Ac-tubulin levels were measured in iPSC-CMs treated with five classes of cardiovascular drugs used as standard of care (SOC) in clinical practice: ARNi, angiotensin receptor neprilysin; ARB, angiotensin II receptor blocker; ACEi, angiotensin-converting enzyme inhibitor; SGLT2, sodium glucose cotransporter 2. Data show that SOC agents do not affect Ac-tubulin levels. [Figure 12B]SOC agents did not significantly affect HDAC6 expression in iPSC-CMs as measured using qPCR. N=2 biological replicates, 4 technical replicates for each experiment per condition. Error bars=SD. [Figure 13A] Figure 2 shows that HDAC6 levels were higher in ischemic versus healthy human hearts. Error bars = SEM. *P<0.05, **P<0.01, ****P<0.0001. [Figure 13B] 1 shows increased levels of HDAC6 in BAG3cKO mice and in a mouse model of heart failure. [Figure 14A] (FIGS. 14A-14J) Inhibition of HDAC6 by TYA-631 protects cardiac function in MLPKO mice. (FIG. 14A) Schematic diagram of drug treatment in the MLPKO mouse model. TYA-631, a selective HDAC6 inhibitor, was administered daily by oral gavage at 30 mg / kg starting when mice were 1.5 months old. [Figure 14B] Immunostaining of iPSC-CMs treated with TYA-631 (5.5 μM) results in high Ac-tubulin. Scale bar = 200 μm. [Figure 14C] The biochemical selectivity of TYA-631 is 3,500-fold more selective for HDAC6 than for other HDACs. [Figure 14D] Western blot of iPSC-CMs treated with TYA-631 stained with monoclonal anti-Ac-lysine. Givinostat (Giv; pan-HDAC inhibitor control) showed both on-target (Ac-tubulin staining) and off-target (Ac-histone H3 and H4 staining) activity. TYA-631 shows only specific on-target activity with no detectable off-target activity. [Figure 14E] Daily TYA-631 dosing preserved cardiac function as measured by ejection fraction over the 9-week dosing period. Error bars = SEM. *P<0.05, **P<0.01. [Figure 14F]Ejection fraction was tracked from the first day of dosing and delta ejection fraction was measured. Mice treated with TYA-631 for 9 weeks showed a 4.0% decrease, while the vehicle-treated group had a 14.8% drop. Error bars = SEM. *P<0.05, **P<0.01. [Figure 14G] Ejection fraction (FIG. 14G) and delta ejection fraction (FIG. 14H) at 15 weeks of age and 9 weeks of treatment (compared to pre-drug baseline) indicate that TYA-631 provides protection from decline in cardiac function in MLPKO mice. Error bars=SEM. **P<0.01. [Figure 14H] Ejection fraction (FIG. 14G) and delta ejection fraction (FIG. 14H) at 15 weeks of age and 9 weeks of treatment (compared to pre-drug baseline) indicate that TYA-631 provides protection from decline in cardiac function in MLPKO mice. Error bars=SEM. **P<0.01. [Figure 14I] Left ventricular diastolic diameter (LVIDd) (FIG. 14I) and left ventricular systolic diameter (LVIDs) (FIG. 14J) were reduced by TYA-631 in MLPKO mice. Error bars = SEM. [Figure 14J] Left ventricular diastolic diameter (LVIDd) (FIG. 14I) and left ventricular systolic diameter (LVIDs) (FIG. 14J) were reduced by TYA-631 in MLPKO mice. Error bars = SEM. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0079] overview The present disclosure generally relates to the demonstration, both in vitro and in vivo, of the efficacy of various HDAC6 inhibitors in dilated cardiomyopathy (DCM). For example, as disclosed herein, both Tubastatin A (more than 100-fold more selective than other HDACs) and TYA-018 (more than 2500-fold more selective than other HDACs) inhibit BAG3 in DCM. cKO and BAG3 E455K In addition, as disclosed herein, TYA-631 inhibits the MLP of DCM. KOIt is effective in mouse models.Furthermore, the efficacy of a number of different HDAC6 inhibitors against HDAC6 is disclosed herein.

[0080] Thus, the present disclosure provides support for the use of HDAC6 inhibitors in the treatment of DCM.

[0081] In some embodiments, provided herein are methods of treating or preventing dilated cardiomyopathy in a subject in need thereof, the methods comprising administering (e.g., orally) an HDAC6 inhibitor to the subject (e.g., human). In some embodiments, provided herein are methods of treating or preventing dilated cardiomyopathy with reduced ejection fraction in a subject in need thereof, the methods comprising administering (e.g., orally) an HDAC6 inhibitor to the subject (e.g., human).

[0082] Advantageously, administration of selective HDAC6 inhibitors may be less toxic than pan-HDAC inhibitors. Without being bound by theory, HDAC6 inhibitors may 1) directly act 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. As sarcomere damage and myofibrillar disorganization are hallmarks of DCM (Dominguez et al., 2018), inhibition of HDAC6 may provide protection at the sarcomere level in DCM.

[0083] definition Unless otherwise indicated by the context, it is specifically intended that the various features of the present invention can 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 composite is described herein as comprising components A, B, and C, it is specifically intended that any of A, B, or C, or combinations thereof, alone or in any combination, may be omitted and discarded.

[0084] 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 by 10%, or alternatively by 5%, or alternatively by 2%. Although not always explicitly stated, it is understood that all numerical designations are preceded by the term "about". It is understood that such range formats are used for convenience and brevity, and should be flexibly understood to include not only the numerical values ​​explicitly 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 explicitly specified. For example, a ratio in the range of about 1 to about 200 should be understood to include the explicitly recited limits of about 1 to about 200, but also to include individual ratios such as about 2, about 3, and about 4, as well as subranges such as about 10 to about 50, about 20 to about 100, etc. It is also understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents thereof are known in the art.

[0085] 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").

[0086] The term "a" or "an" may refer to one or more of that entity, i.e., may refer to a plural reference. Thus, the terms "a", "an", "one or more", and "at least one" are used interchangeably herein. In addition, reference to "an element" by the indefinite article "a" or "an" does not exclude the possibility that more than one element is present, unless the context clearly requires that only one of the element is present.

[0087] 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 among samples 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 exceeds 100% or falls below 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.

[0088] As used herein, the term "HDAC6" refers to the enzyme encoded by the HDAC6 gene in humans.

[0089] As used herein, the term "HDAC6 inhibitor" refers to a compound that inhibits at least one enzymatic activity of HDAC6.

[0090] 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 over HDAC1 is about 5 to about 30,0000, e.g., 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.

[0091] 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 HDAC6 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.

[0092] 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 its symptoms.

[0093] As used herein, the term "prevention" 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.

[0094] "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 can 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.

[0095] As used herein, the term "effective amount" refers to an amount 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 a number of 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 depends 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.

[0096] 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" can be a mammal, such as a dog, a cat, a horse, a farm animal, a zoo animal, or a human. A subject or patient can also be any domestic animal, such as a bird, a pet, or an industrial animal. Specific examples of "subject" and "patient" include, but are not limited to, individuals with a cardiac disease or disorder, and individuals with a characteristic or symptom associated with a cardiac disorder.

[0097] The phrase "pharmacologically acceptable" is used herein to refer to compounds, materials, compositions, and / or dosage forms that are suitable, within the scope of sound medical judgment, 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.

[0098] The term "pharmaceutical acceptable salts" includes salts obtained by reacting an active compound that functions as a base with an inorganic or organic acid to form a salt, such as, 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.

[0099] "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 group that is C1-C 12 Alkyl, containing up to 10 carbon atoms, is C1-C 10 An alkyl having up to 6 carbon atoms is a C1-C6 alkyl, and an alkyl having up to 5 carbon atoms is a 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 12Includes alkyl. C1~C 12 Non-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.

[0100] "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.

[0101] "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 those having C2 to C4. 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 mentioned above for C2-C5 alkenyl, but also includes C6 alkenyl. C2-C 10Alkenyl includes all of the moieties described above for C2-C5 alkenyl and C2-C6 alkenyl, but also includes C7, C8, C9, and C 10 Alkenyl is also included. 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 , 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.

[0102] "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. 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., as 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.

[0103] "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 group 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.

[0104] "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 can be through any two carbons in the chain having suitable valences. Unless otherwise stated in this specification, the alkynylene chain can be optionally substituted.

[0105] "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.

[0106] "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 herein, an aryl may be optionally substituted.

[0107] "Carbocyclyl", "carbocyclic ring", or "carbocycle" refers to a ring structure in which each of the atoms forming the ring is carbon and is attached to the remainder of the molecule by a single bond. A carbocyclic ring can contain from 3 to 20 carbon atoms in the ring. Carbocyclic rings include aryl, as well as cycloalkyl, cycloalkenyl, and cycloalkynyl, as defined herein. Unless otherwise stated in the specification, a carbocyclyl group can be optionally substituted.

[0108] 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.

[0109] "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.

[0110] "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.

[0111] "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.

[0112] "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.

[0113] "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 are attached to the remainder of the molecule by single bonds. Heterocyclyl or heterocyclic rings include 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.

[0114] "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 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, indazolyl, indolyl, indolyl, and indolyl. Examples include, but are not limited to, dazolyl, 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.

[0115] "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 heterocyclylalkyl group may be optionally substituted.

[0116] 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; oxygen atoms in groups such as hydroxyl, alkoxy, and ester groups; sulfur atoms in groups such as thiol, thioalkyl, sulfone, sulfonyl, and sulfoxide groups; nitrogen atoms in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; silicon atoms 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 are replaced by a higher order bond (e.g., a double bond or a 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 are replaced by a bond such as -NR g R h , -NR g C(=O)R h , -NRg C(=O)NR g 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 includes any of the above groups, where 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 are 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 in which 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 substituents.

[0117] 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 3is R 3 If is "XY", the bond point is R 3 is assumed to be the same bond depicted as being attached to CH3.

[0118] As used herein, the term "ameliorate" refers to increasing the level of a biochemical or physiological parameter to a level observed in a subject prior to the onset of a disease or condition, or to a level observed in a subject not having the disease or condition.

[0119] As used herein, the term "reduce" refers to decreasing the level of a biochemical or physiological parameter.

[0120] As used herein, the term "cardiomyopathy" refers to any disease or dysfunction of the myocardium (heart muscle) in which the heart becomes abnormally enlarged, thickened, and / or stiffened. As a result, the ability of the heart muscle to pump blood is usually weakened. The etiology of the disease or disorder can be, for example, inflammatory, metabolic, toxic, infiltrative, fibrogenic, hematological, genetic, or of unknown origin. There are two general types of cardiomyopathies: ischemic (caused by lack of oxygen) and non-ischemic.

[0121] As used herein, the term "dilated cardiomyopathy" or "DCM" refers to a condition in which the heart muscle weakens and enlarges. As a result, the heart cannot pump enough blood to the rest of the body. The most common causes of dilated cardiomyopathy are heart diseases caused by narrowing or blockage of coronary arteries, uncontrolled high blood pressure, alcohol or drug abuse, diabetes, thyroid disease, or hepatitis, side effects of medications, abnormal heart rhythms, autoimmune diseases, genetic causes, infections, narrowed or leaky heart valves, pregnancy, and exposure to heavy metals such as lead, arsenic, cobalt, or mercury. DCM can affect people of any age. However, it is most common in adult males. DCM includes idiopathic DCM. In some embodiments, the DCM is familial DCM.

[0122] 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.

[0123] Heart failure is a complex clinical syndrome that can result from any structural or functional cardiovascular disorder caused by insufficient systemic perfusion to meet the metabolic demands of the body without excessively increasing left ventricular filling pressure. It is characterized by specific symptoms such as dyspnea and fatigue, as well as signs such as fluid retention.

[0124] 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 overweight. CHF is broadly classified as HF with reduced or preserved ejection fraction (HFrEF and HFpEF) according to the contractile function of the left ventricle. The term "heart failure" does not mean that the heart has stopped or is not functioning completely, but that it is weaker than normal in a healthy person. 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, swelling of the legs and ankles, chest pain, and coughing. In some embodiments, the methods of the present disclosure reduce, prevent, or ameliorate one or more symptoms of heart failure in a subject suffering from or at risk for heart failure associated with DCM.

[0125] As used herein, the term "detrimental mutation" refers to a mutation that reduces the function of a gene. Detrimental mutations may 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 deletion of a gene. As used herein, the term may refer to homozygous or heterozygous mutations in a gene where the mutation has an phenotypic effect in the carrier.

[0126] As used herein, the term "left ventricular internal diameter diastolic" or "LVIDd" refers to the size of the left ventricle during diastole.

[0127] As used herein, the term "left ventricular systolic dimensions" or "LVIDs" refers to the size of the left ventricle during systole.

[0128] As used herein, the term "left ventricular mass" refers to the weight of the left ventricle.

[0129] 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.

[0130] The detailed description of the present disclosure is divided into sections for the convenience of the reader only, and the disclosure found in any section can be combined with the disclosure in 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 can also be used in the practice or testing of the present invention, the preferred methods and materials are described below.

[0131] Treatment or prevention of dilated cardiomyopathy (DCM) Methods for treating or preventing dilated cardiomyopathy using HDAC6 inhibitors are provided.

[0132] Familial DCM In some embodiments, DCM is familial DCM. There are various known cases of familial DCM. The genes involved in familial DCM can include TTN, DSP, MYBPC3, SCN5A, RBM20, LDB3, LMNA, ANKRD1, MYH7, TNNT2, BAG3, DMD, MYPN, CSRP3 (known as MLP), MYH6, TNNI3, ABCC9, TPM1, PSEN2, DES, or MYOZ2.

[0133] BAG3 One gene that is essential for maintaining control of protein quality is BCL2-associated athanogene3 (BAG3). BAG3 is a stress response gene that acts as an HSP70 chaperone cofactor in a complex with small heat shock proteins (HSPs) to maintain cardiomyocyte function (Franceschelli et al., 2008; Judge et al., 2017; Rauch et al., 2017). BAG3 is highly expressed in cardiac and skeletal muscles and can localize to Z-discs (Homma et al., 2006). BAG3 has also been proposed to protect muscle cells from mechanical injury and proteotoxic stress (Dominguez et al., 2018; Judge et al., 2017).

[0134] Mutations in BAG3 are associated with DCM. In adults over 40 years of age, loss-of-function BAG3 mutations show 80% penetrance for DCM (Dominguez et al., 2018). Familial BAG3 mutations are autosomal dominant, suggesting a heterozygous loss-of-function mechanism (Chami et al., 2014; Judge et al., 2017; Villard et al., 2011). BAG3 mutations resulting in loss of function account for approximately 3% of the variant distribution in the DCM gene (Haas et al., 2015). Most mutations in BAG3 are deleterious (e.g., E455K), but cardioprotective variants (C151R) have also been reported (Villard et al., 2011). This finding suggests that the BAG3 chaperone complex may acquire a gain-of-function phenotype that protects against proteotoxic stress and mechanical injury in the heart. In addition, mutations in BAG3 resulted in cardiac-related phenotypes in both in vivo and in vitro models, including zebrafish (Norton et al., 2011; Ruparelia et al., 2014), mice (Fang et al., 2017; Homma et al., 2006), and human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) (Judge et al., 2017). Also, adequate BAG3 levels are required to maintain chaperone function and maintain protein quality control, as reduction of BAG3 was observed in patients with idiopathic DCM (Feldman et al., 2014). Thus, BAG3 is an attractive target for developing novel small molecule therapeutics for BAG3 myopathy. These efforts may also lead to interventions for other genetic causes of DCM and non-genetic forms of heart failure (Sturner and Behl, 2017).

[0135] Most mutations in BAG3 are deleterious (e.g., E455K), but a cardioprotective variant (C151R) has also been reported.

[0136] Nonfamilial DCM In some embodiments, the DCM is non-familial DCM, including but not limited to idiopathic DCM, in some embodiments, the DCM is drug-induced cardiomyopathy (e.g., due to anti-cancer therapy or anti-retroviral therapy), viral myocarditis, or postpartum cardiomyopathy.

[0137] HDAC6 inhibitors Histone deacetylases ("HDACs") are a class of enzymes that have deacetylase activity with a wide range of genomic and non-genomic substrates. Based on sequence identity and catalytic activity, 11 zinc-dependent HDAC enzymes have been classified (Haberland et al., 2009).

[0138] 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 controlling 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. In addition, significant adverse effects of pan-HDAC inhibitors (e.g., SAHA and Panabinostat), including fatigue, nausea, diarrhea, and thrombocytopenia, have been observed in clinical settings (Subramanian et al., 2010).

[0139] In the cardiac indication setting, most studies have utilized pan-HDAC inhibitors (e.g., SAHA, TSA, and Givinostat) to treat pressure overload rodent models including transcatheter aortic arch coarctation (TAC) (Cao et al., 2011), hypertension in Dahl salt-sensitive rats (Jeong et al., 2018), and myocardial infarction (Nagata et al., 2019). In addition, HDAC6-selective inhibitors have been used to ameliorate the effects of pressure overload in rodent models (Demos-Davies et al., 2014) and provide protection against proteotoxicity in transgenic mouse models of cardiomyopathy (McLendon et al., 2014). However, these experiments in pressure overload rodent models are not predictive of treating dilated cardiomyopathy. Pressure overload in adult mice induces cardiomyocyte hypertrophy through increased cardiomyocyte size, enhanced protein synthesis, and new sarcomere assembly. Mohammadi et al. Nature Protocols 16:775-790 (2021). Pressure overload is a model of physiological exogenous injury to otherwise normal cardiomyocytes. Dilated cardiomyopathy results from an intrinsic defect to cardiomyocytes (e.g., mutations in genes associated with cardiac function). Furthermore, pressure overload models hypertrophic disease rather than myocardial frailty in dilated cardiomyocytes.

[0140] 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, and its best-characterized substrates include tubulin, HSP90, and cortactin (Brindisi et al., 2019).

[0141] Pharmacological inhibition of HDAC6 blocks its deacetylase activity, leading to hyperacetylation of its substrates, particularly tubulin ( Hubbert et al., 2002 ).

[0142] 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 ).

[0143] Hydroxyl acids are zinc chelators and have been widely used in the development of pan- and HDAC-selective inhibitors. However, most hydroxamic acid HDAC inhibitors lack the desired selectivity or exhibit poor pharmacokinetic profiles and poor bioavailability (Butler et al., 2010; Santo et al., 2012).

[0144] A variety of selective HDAC6 are known in the art.In addition, it is common to use known methods to screen compounds and identify further selective HDAC6 inhibitors.In particular, taking into account known HDAC6 inhibitors, those skilled in the art can identify which analogues of compounds have selective HDAC6 activity.

[0145] In some embodiments, the HDAC6 inhibitor is a gene silencing agent, such as an RNA silencing agent (eg, siRNA).

[0146] 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]

[0147] More examples of HDAC6 inhibitors, U.S. Patent Publication No. US8227516(B2), No. US2010 / 0292169(A1), and the same No. US2007 / 0207950(A1), same as US8222423(B2), same as US2010 / 0093824(A1), same as U.S. S2010 / 0216796(A1), same as US8673911(B2), same as US8217076(B2), same as US8440716(B2), same as US2011 / 0195432(A1), same as US8624040(B2), same as US9096518(B2) , same as US8431538(B2), same as US2012 / 0258993(A1), same as US8546588(B2), same as US8513421(B2), same as US2014 / 0031368(A1), US2012 / 0015943(A1), US2012 / 0015942(A1), same as US2014 / 0243335(A1), same as US2013 / 0225543(A1), same as US8471026(B2), same as US9238028(B2), same as US8765773(B2), same as USRE47009(E1), same as US2014 / 0294856(A1), same as US9512083(B2), same as US9670193(B2), same as US9345905(B2), same as US9409858(B2), same as US9663825(B2), same as US2015 / 0119327(A1), same as US2015 / 0250786(A1), same as US10041046(B2), same as US9586973(B2), same as US2016 / 0069887(A1), same as US2014 / 0357512(A1), same as US9751832(B2), same as US2 016 / 0228434(A1), same as US2015 / 0105358(A1), same as US10660890(B2), same as US2016 / 0271083(A1), same as US2015 / 0176076(A1), same as US2020 / 0405716(A1), same as US9890136(B2), same as US10287255(B2), same as US2017 / 0173083(A1), same as US10016421(B2), same as US9987258(B2), same as US10568854(B2), same as US10106540(B2),US10266489(B2), US9993459(B2), US10183934(B2), US10494354(B2), US10494353(B2), US10112915(B2), US10377726 (B2), US10829462(B2), US10829461(B2), US2021 / 0009539(A1), US2021 / 0009538(A1), US10239845(B2), US10472337(B2) No. US10479772(B2), US10464911(B2), US10584117(B2), US10538498(B2), US10011611(B2), US10494355(B2), US10040 769(B2), US10858323(B2), US10654814(B2), US2019 / 0209559(A1), US2019 / 0185462(A1), US2019 / 0192521(A1), US2019 / 0321361(A1), US2020 / 0046698(A1), US2019 / 0262337(A1), US2019 / 0282573(A1), US2019 / 0282574(A1), US2020 / 0071288(A 1), US10745389(B2), US10357493(B2), US2020 / 0171028(A1), US2020 / 0054773(A1), US2020 / 0308174(A1), US2020 / 01555 49(A1), US10435399(B2), US2020 / 0216563(A1), US2019 / 0216751(A1), US2020 / 0339569(A1), US2021 / 0078963(A1), US2021 / 0077487(A1), US2019 / 0270733(A1), US2019 / 0270744(A1), US2020 / 0022966(A1), and US2021 / 0094944(A1),Incorporated herein for the 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.

[0148] 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 entire contents of which are incorporated herein by reference.PCT / US2020 / 066439, published as WO2021 / 127643(A1), also describes the synthesis method of such compounds, which is specifically incorporated herein by reference.

[0149] In some embodiments, the HDAC6 inhibitor is a compound of formula (II): [ka] R 1 is selected from the group consisting of: [Table 12] 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 R5 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, together with the atoms to which they are attached, they form a cycloalkyl or heterocyclyl, Y is CR 2 or N, n is selected from 0, 1, and 2.

[0150] 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.

[0151] 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 6 In 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. In some embodiments, X 1 is S or NR 6 In some embodiments, R 6 is C1-C6 alkyl.

[0152] In some embodiments of Formula (I), R 2 and R 3 is H.

[0153] 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.

[0154] 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 R4 and R 5 together with the atom to which they are attached form a cycloalkyl or heterocyclyl, each of which is optionally substituted.

[0155] 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.

[0156] In some embodiments of Formula (I), R 5is aryl, heteroaryl, or cycloalkyl, each of which is optionally substituted.

[0157] In some embodiments, R 5 is aryl. In some embodiments, aryl is [ka] where R b is one or more selected from the group consisting of halogen, haloalkyl, alkyl, Oalkyl, Ohaloalkyl, alkylene-Ohaloalkyl, cycloalkyl, heterocyclyl aryl, heteroaryl, alkylnitrile, or CN. In some embodiments, haloalkyl is selected from CF3, CF2CH3, CHF2, or CH2F. 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, 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.

[0158] 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 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, heterocyclyl aryl, heteroaryl, alkylnitrile, or CN. In some embodiments, haloalkyl is selected from CF3, CF2CH3, CHF2, or CH2F. 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, 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.

[0159] 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.

[0160] 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.

[0161] In some embodiments of Formula (I), R 5 is optionally substituted with one or more halogen, haloalkyl, alkyl, Oalkyl, Ohaloalkyl, cycloalkyl, heterocyclyl aryl, 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.

[0162] 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 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.

[0163] 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 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 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.

[0164] 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 5 is 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 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 5-6 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, C1~6 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 C4-6 cycloalkyl. 1~6 Haloalkyl is CF, CHF, or CHF. In some embodiments, OC 1~6 Haloalkyl 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.

[0165] 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.

[0166] 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.

[0167] In some embodiments of Formula (I), R 1 teeth, [ka] is selected from the group consisting of:

[0168] 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]

[0169] 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 1~3 Alkylalkyl is methyl, ethyl, or isopropyl. In some embodiments, haloalkyl is CF3, CHF2, or CH2F.

[0170] In some embodiments of Formula (I), Y is CH and R 4 and R 5 is H.

[0171] 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.

[0172] 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.

[0173] 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.

[0174] 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.

[0175] In some embodiments, the disclosure provides a compound of formula (Ia) or a pharma- ceutically acceptable salt thereof: [ka]

[0176] R 1 , R 2 , R 3 , R 4 , R 5 , R a , X 1 , n, and Y are as defined above for formula (I).

[0177] 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 R 2 , R 3 , R 4 , R 5 , and R a is as defined above for formula (I).

[0178] 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, where the heteroaryl is an optionally substituted pyridine or pyrazine. In some embodiments, the heteroaryl is an optionally substituted pyridine.

[0179] 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 a is H. In some embodiments, R 5 is an optionally substituted pyridine.

[0180] 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; ais 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.

[0181] 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 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.

[0182] 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 4is -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.

[0183] In some embodiments, the disclosure provides a compound of formula (Ib) or a pharma- ceutically acceptable salt thereof: [ka] R 1 , R 2 , R 3 , R 4 , R 5 , R a , X 1 , n, and Y are as defined above for formula (I).

[0184] 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.

[0185] In some embodiments of Formula (I)-(Ib), each optionally substituted haloalkyl independently is 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.

[0186] 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.

[0187] 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.

[0188] In some embodiments of Formula (I)-(Ib), each optionally substituted aryl is independently C 6~12 In a further embodiment, C 6~12 Aryl is optionally substituted phenyl.

[0189] In some embodiments of Formulae (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.

[0190] In some embodiments, the compound of formula (I) is selected from the group consisting of: [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6]

Table 5-7

Table 5-8

Table 5-9

Table 5-10

Table 5-11

Table 5-12

Table 5-13

Table 5-14

Table 5-15

Table 5-16

Table 5-17

Table 5-18

Table 5-19

Table 5-20

Table 5-21

Table 5-22

Table 5-23

[0191] 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).

[0192] In some embodiments of Formula (Ic), R a is H. In some embodiments, R a is F. In some embodiments, R a is Me.

[0193] 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.

[0194] 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, heterocyclyl aryl, heteroaryl, alkylnitrile, or CN.b is 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.

[0195] 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 groups, together with the carbon atoms to which they are attached, may form bridged or fused C 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.

[0196] In some embodiments, the 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 groups, together with the carbon atoms to which they are attached, may form bridged or fused C 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.

[0197] 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 groups, together with the carbon atoms to which they are attached, may form bridged or fused C 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.

[0198] 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 groups, together with the carbon atoms to which they are attached, may form bridged or fused C 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.

[0199] In some embodiments, the compound has the formula: [ka] During the ceremony, 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.

[0200] 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.

[0201] 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).

[0202] 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.

[0203] 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’ ).

[0204] 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 groups, together with the carbon atoms to which they are attached, may form bridged or fused C 3~7 In some embodiments, two R c The groups, together with the carbon atoms to which they are attached, may be optionally substituted bridged or fused C 3~7 In some embodiments, two R cGroups, taken together with the carbon atoms to which they are attached, form an optionally substituted bridged or fused 5- or 6-membered heterocyclyl. In 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 The alkyl is isopropyl.

[0205] 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 1~5 In some embodiments, -C1~5 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.

[0206] 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.

[0207] 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.

[0208] 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.

[0209] 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.

[0210] 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.

[0211] In some embodiments, the 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.

[0212] In some embodiments, the 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 is 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.

[0213] 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 is H, alkyl, -C(O)R e , sulfonyl, cycloalkyl, aryl, or heteroaryl; R eand 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.

[0214] In some embodiments of Formulas (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.

[0215] In some embodiments of formulas (Ih)-(Ij), X 1 , X 2 , X 3 , and X 4 Each of X is CH. 1 , X 2 , X 3 , and X 4 is N. In some embodiments, one of X 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. 2 is N and X 1 , X 3 , and X 4 Each of X is CH. 3 is N and X1 , X 2 , and X 4 Each of X is CH. 4 is N and X 1 , X 2 , and X 3 Each of the is CH.

[0216] 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 In some embodiments, U is CH and X 2 is N and X 1 , X 3 , and X 4 In some embodiments, U is CH and X 3 is N and X 1 , X 2 , and X 4 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.

[0217] 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 4 In some embodiments, U is O and X2 is N and X 1 , X 3 , and X 4 In some embodiments, U is O and X 3 So, N and X. 1 , X 2 , and X 4 In some embodiments, U is O and X 4 is N and X 1 , X 2 , and X 3 Each of the is CH.

[0218] 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.

[0219] 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.

[0220] 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 The groups are attached to the same carbon atom, which may also be referred to as a germinal substitution. c The groups, together with the atoms to which they are attached, are optionally substituted C 3~6 In some embodiments, two R c The groups taken together with the atom to which they are attached form an optionally substituted cyclopropyl. In some embodiments, the optional substituents are one or more of R bIn 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 is also referred to as germinal substitution.

[0221] 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.

[0222] 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).

[0223] 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 Groups, together with the carbon atom to which they are attached, may be optionally substituted C 3~6 In some embodiments, two R c The groups, taken together with the carbon atom to which they are attached, form cyclopropyl or cyclobutyl, each of which is optionally substituted. In some embodiments, 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 is also referred to as germinal substitution.

[0224] 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, i-propyl, or t-butyl. In some embodiments, cycloalkyl is cyclopropyl, cyclopentyl, or cyclohexyl.

[0225] 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.

[0226] 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.

[0227] 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.

[0228] In some embodiments, the HDAC6 inhibitor has the following 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 taken together with the atom to which they are attached form a heterocyclyl, each of which is optionally substituted.

[0229] In some embodiments, R a is H.

[0230] In some embodiments, R 1 is as follows: [ka]

[0231] In some embodiments, R 4 is -(SO2)R 2 It is.

[0232] In some embodiments, -(SO)R 2is -(SO2)alkyl, -(SO2)alkyleneheterocyclyl, -(SO2)haloalkyl, -(SO2)haloalkoxy, or -(SO2)cycloalkyl.

[0233] In some embodiments, R 5 is heteroaryl.

[0234] In some embodiments, the heteroaryl is a 5-6 membered heteroaryl.

[0235] 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.

[0236] In some embodiments, R b are F, Cl, -CH3, -CH2CH3, -CF3, -CHF2, -CF2CH3, -CN, -OCH3, -OCH2CH3, -OCH(CH3)2, -OCF3, -OCHF2, -OCH2CF2H, and cyclopropyl.

[0237] 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.

[0238] In some embodiments, the HDAC6 inhibitor is represented by 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.

[0239] In some embodiments, R b is H, halogen, haloalkyl, or haloalkoxy.

[0240] In some embodiments, R 4 is optionally substituted alkyl or cycloalkyl.

[0241] 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.

[0242] In some embodiments, R b is H, halogen, haloalkyl, or haloalkoxy.

[0243] In some embodiments, R 4 is optionally substituted alkyl or cycloalkyl.

[0244] In some embodiments, R 4 is alkyl.

[0245] 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.

[0246] In some embodiments, R b is H, halogen, haloalkyl, or haloalkoxy.

[0247] In some embodiments, R 4 is optionally substituted alkyl.

[0248] In some embodiments, the HDAC6 inhibitor is a compound having the formula [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 taken together with the atom to which they are attached form a heterocyclyl, each of which is optionally substituted.

[0249] In some embodiments of formula I(y), R a is H.

[0250] In some embodiments of formula I(y), R 1 teeth, [ka] It is.

[0251] In some embodiments of formula I(y), R 4 is -(SO2)R 2 It is.

[0252] In some embodiments of Formula I(y), -(SO2)R 2 is -(SO2)alkyl, -(SO2)alkyleneheterocyclyl, -(SO2)haloalkyl, -(SO2)haloalkoxy, or -(SO2)cycloalkyl.

[0253] In some embodiments of formula I(y), R 5 is heteroaryl.

[0254] In some embodiments of Formula I(y), the heteroaryl is a 5-6 membered heteroaryl.

[0255] 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.

[0256] 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.

[0257] 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.

[0258] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0259] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0260] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0261] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[0262] In some embodiments, the HDAC6 inhibitor has the structure: [ka]

[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 is TYA-018 or an analog thereof. The structure of TYA-018 is: [ka]

[0269] Analogs of TYA-018 include, but are not limited to, the compounds listed in Table 2. [Table 2-1] [Table 2-2]

[0270] 5-Fluoronicotinamide Derivatives In some embodiments, the HDAC6 inhibitor is a 5-fluoronicotinamide derivative. Exemplary 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 entire contents of which are incorporated herein by reference. PCT / US2020 / 054134, published as WO2021 / 067859(A1), also describes the synthesis method of such compounds, which is specifically incorporated herein by reference.

[0271] In some embodiments, the HDAC6 inhibitor is a compound of formula (II): [ka] 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; 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; 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.

[0272] Pharmaceutical Compositions and Kits In various embodiments of the present disclosure, a pharmaceutical composition is provided that includes 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 the composition or formulation for various purposes. In some embodiments, a pharmaceutical composition that includes one or more compounds disclosed herein, or a pharma- ceutical acceptable solvate, hydrate, tautomer, N-oxide, or salt thereof, further includes a pharma- ceutical acceptable carrier. In some embodiments, the pharma- ceutical acceptable carrier includes a pharma- ceutical acceptable excipient, binder, and / or diluent. In some embodiments, suitable pharma- ceutical acceptable excipients include, but are not limited to, water, salt solution, alcohol, polyethylene glycol, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, and polyvinylpyrrolidone.

[0273] 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)).

[0274] In another aspect, the disclosure provides an HDAC6 inhibitor for use in a method for treating dilated cardiomyopathy.

[0275] In another aspect, the disclosure provides a kit comprising an HDAC6 inhibitor or a pharmaceutical composition thereof and instructions for use in a method for treating dilated cardiomyopathy.

[0276] In another aspect, the disclosure provides the use of an HDAC6 inhibitor in the treatment of dilated cardiomyopathy.

[0277] Screening Methods In another aspect, the disclosure provides a method for identifying a compound for the treatment of dilated cardiomyopathy, the method comprising contacting a cell culture comprising cells having an inactivating mutation in BAG3 with each member of a plurality of candidate compounds, and selecting a compound that reduces sarcomere damage in the cells. In another aspect, the disclosure provides a method for identifying a compound for the treatment of dilated cardiomyopathy, the method comprising contacting a cell culture comprising cells having an inactivating mutation in MLP (CSRP3) with each member of a plurality of candidate compounds, and selecting a compound that reduces sarcomere damage in the cells.

[0278] In another aspect, the disclosure provides a method of treating dilated cardiomyopathy in a subject in need thereof, the method comprising contacting a cell culture comprising cells having an inactivating mutation in BAG3 with each member of a plurality of candidate compounds, selecting a selected compound as one that reduces sarcomere damage, and administering a therapeutically effective amount of the selected compound to the subject. In another aspect, the disclosure provides a method of treating dilated cardiomyopathy in a subject in need thereof, the method comprising contacting a cell culture comprising cells having an inactivating mutation in MLP (CSRP3) with each member of a plurality of candidate compounds, selecting a selected compound as one that reduces sarcomere damage, and administering a therapeutically effective amount of the selected compound to the subject.

[0279] Method of Administration and Patient Population Treated 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.

[0280] 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.

[0281] A variety of dosing 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.

[0282] 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 orally once a day.

[0283] 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.

[0284] 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.

[0285] 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 the 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).

[0286] 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. In some embodiments, one or more of the HDAC6 inhibitors described herein (and pharmaceutical compositions comprising such HDAC6 inhibitors) can 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, a therapy for a cardiac condition (e.g., heart failure), and / or a therapy for DCM. This additional therapy can be any cardioprotective therapy, a cardiac condition therapy (e.g., heart failure) therapy, or an anti-DCM therapy known in the art. In some embodiments, the HDAC6 inhibitors described herein (and pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with another anti-DCM therapy. In some embodiments, the HDAC6 inhibitors described herein (and pharmaceutical compositions comprising such HDAC6 inhibitors) are administered to a subject in combination with a cardioprotective therapy. 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 before, simultaneously with, or after an additional therapy (cardioprotective therapy or anti-DCM therapy, such as an ACE inhibitor or beta-blocker). In some embodiments, the subject treated according to the methods described herein is not receiving anti-DCM therapy, cardioprotective therapy, and / or cardiac condition (e.g., heart failure) therapy.

[0287] 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 treating DCM or a cardioprotectant). 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, such as an ACE inhibitor, a beta blocker, or another agent for treating DCM or for cardioprotection (e.g., in a therapeutically effective amount).

[0288] In some embodiments, the subject is a human. In some embodiments, the human is an adult human. In some embodiments, the subject is a male. In some embodiments, the subject is a female.

[0289] In some embodiments, the subject has cardiomyopathy (e.g., has symptoms of or has been diagnosed with cardiomyopathy). In some embodiments, the cardiomyopathy is a genetic cardiomyopathy. In some embodiments, the cardiomyopathy is a non-genetic cardiomyopathy. In some embodiments, the subject has DCM (e.g., has symptoms of or has been diagnosed with DCM). In some embodiments, the subject has familial DCM (e.g., has symptoms of or has been diagnosed with familial DCM). In some embodiments, the subject has non-familial DCM (e.g., has symptoms of or has been diagnosed with non-familial DCM). In some embodiments, the subject has idiopathic DCM (e.g., has symptoms of or has been diagnosed with idiopathic DCM). In some embodiments, the subject has DCM with reduced ejection fraction (e.g., has symptoms of or has been diagnosed with DCM with reduced ejection fraction).

[0290] In some embodiments, the subject has a deleterious mutation in BAG3 (e.g., a mutation that results in a deletion of BAG3 or inactivation of BAG3). In some embodiments, the subject has a deleterious mutation in MLP, also known as CSRP3 (e.g., a mutation that results in a deletion of MLP or inactivation of MLP).

[0291] Numbered Embodiments of the Invention 1. A method for treating or preventing dilated cardiomyopathy in a subject in need thereof, comprising administering an HDAC6 inhibitor to the subject. 2. The method of embodiment 1, wherein the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative. 3. The method of embodiment 2, wherein the HDAC6 inhibitor is a fluoroalkyl-oxadiazole derivative according to the following general formula: [ka] 4. The HDAC6 inhibitor is a compound according to formula (I) [ka] or a pharma- ceutically acceptable salt thereof, R 1 is selected from the group consisting of: [Table 7] R a But, 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 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 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 together with the atom to which they are attached form a cycloalkyl or heterocyclyl, Y is CR 2 or N, 2. The method of embodiment 1, wherein n is selected from 0, 1, and 2. 5. The method of embodiment 4, wherein the HDAC6 inhibitor is selected from the group consisting of: [Table 8-1] [Table 8-2] 6. The method of embodiment 4, wherein the HDAC6 inhibitor is selected from the group consisting of: [Table 9-1] [Table 9-2]

Table 9-3

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[0292] The present invention is further illustrated by the following examples, which are non-limiting and merely representative of various aspects of the present invention. Solid and dotted wedges in structures disclosed herein indicate relative stereochemistry; absolute stereochemistry is indicated only where specifically stated or depicted.

[0293] Example 1 overview To identify candidate therapeutics, we developed an in vitro DCM model using induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) depleted in BAG3. These BAG3-deficient iPSC-CMs allowed us to identify cardioprotective drugs using phenotypic screening and deep learning (Figure 1). Using a library of 5500 bioactive compounds and siRNA validation, we identified that inhibiting HDAC6 was cardioprotective at the sarcomere level. We repeated this finding in a BAG3 cardiac knockout (BAG3 cKO ) mouse model and show that inhibition of HDAC6 with two isozyme selective inhibitors (Tubastatin A and TYA-018) protects cardiac function. TYA-018 is a compound within formula (I), as well as compounds within formula I(y) and formula (Ic).

[0294] HDAC6 inhibitors suppress BAG3 in DCM cKO In mouse models, HDAC6 inhibitors improved left ventricular ejection fraction and extended life span, and protected the microtubule network from mechanical injury, increased autophagic flux, reduced apoptosis, and reduced inflammation in the heart.

[0295] This example shows that HDAC6 inhibitors can successfully treat subjects with dilated cardiomyopathy. Significantly, HDAC6 inhibitors can treat dilated cardiomyopathy (Figures 11B-11C) and significantly reduce LVIDd and LVIDs (Figures 11D-6E) as measured by EF.

[0296] result An in vitro model of DCM was to transfect cardiomyocytes derived from induced pluripotent stem cells (iPSC-CMs) with siRNA for BAG3. Reduced expression of MYBPC3 and p62 (Figure 2A) and visually scored sarcomere damage (Figure 2B) confirmed the presence of BAG3. KD It is confirmed that the iPSC-CMs recapitulate the phenotype of cardiomyocytes in subjects suffering from or at risk for DCM.

[0297] BAG3 KD To efficiently and reproducibly quantify sarcomere damage in iPSC-CMs, we employed an image analysis method using deep learning (LeCun et al., 2015). We developed a two-class deep learning model based on healthy (SCR siRNA-treated) and diseased (BAG3 siRNA-treated) iPSC-CMs.

[0298] 5,500 bioactive compounds were screened. iPSC-CMs were seeded, allowed to recover, and treated with either SCR or BAG3 siRNA, followed by treatment with bioactive compounds at a concentration of 1 μM (Figure 3A). Deep learning was used to determine a sarcomere score for each compound. A high sarcomere score indicated low levels of sarcomere damage, a low sarcomere score indicated high sarcomere damage, and a negative sarcomere score indicated the compound was toxic (Figure 3B).

[0299] Screening results were ranked based on sarcomere scores and a hit threshold of 0.3 (1% false discovery rate) was assigned. Results from wells treated with either SCR or BAG3 siRNA were plotted as controls. These data showed that cells treated with SCR siRNA had sarcomere scores ranging from 0.3 to 1, while cells treated with BAG3 siRNA had sarcomere scores below 0.3. After manual exclusion of false positive hits (due to rare staining and imaging artifacts), the top 24 hits from the screen were grouped into distinct target classes (Figure 3C).

[0300] The top predicted cardioprotective compounds were categorized into two major classes: HDAC inhibitors and microtubule inhibitors. Of these, the screen identified three compounds that could be broadly categorized as "standard of care" agents for cardiovascular indications: omecamtivmecarbil (a cardiac myosin activator), sotalol (a beta and K channel blocker), and anagrelide (a PDE3 inhibitor). These results further validate the translational relevance of using iPSC-CMs and deep learning to identify cardioprotective compounds in an unbiased, high-throughput manner.

[0301] The top compounds identified in the primary screen included 2Pm IC 50 and CAY10603, a potent and selective HDAC6 inhibitor with over 200-fold selectivity over other HDACs.

[0302] Given the abundance of hit enrichment with HDAC inhibitors, we hoped to ensure that HDAC inhibitors did not prevent sarcomere damage by increasing BAG3 expression in wild-type (WT) iPSC-CMs. To ensure broad-spectrum inhibitors were captured, we used all HDAC inhibitors identified from the screen, plus additional HDAC inhibitors. Using immunostaining and QPCR, we found that none of the HDAC inhibitors increased BAG3 expression in WT iPSC-CMs (Figures 4A-4B). These data suggest that HDAC inhibitors did not provide protection from sarcomere damage by preventing BAG3 knockdown or upregulation of BAG3. Instead, they confer cardioprotection through a different mechanism.

[0303] HDAC6 inhibition provides protection from BAG3 loss of function in iPSC-CMs Secondary validation of the top hits from the primary screen was performed and the results are summarized in Figure 5A. These results highlighted HDAC and microtubule inhibitors as putative cardioprotective compounds. HDAC inhibitors show different levels of polypharmacology for different HDAC isozymes. For example, I class HDACs (HDAC1, 2, 3, and 8) are mainly located in the nucleus and target histone substrates. Inhibiting these isozymes activates global or specific gene expression programs (Haberland et al., 2009). We further investigated all HDACs individually using siRNA to co-knockdown BAG3 and individual HDAC isoforms (HDAC1-HDAC11). In independent studies (2-7 biological replicates), co-knockdown of HDAC6 and BAG3 prevented sarcomere damage induced by BAG3 knockdown as measured by cardiomyocyte score (Figure 5B). Representative immunostaining of BAG3 siRNA-treated cells showed damaged sarcomeres, which appeared to be significantly reduced by knockdown of HDAC6 (FIG. 5C).

[0304] These findings were further validated using siRNAs targeting HDAC1 to HDAC11 independently. Two siRNAs (1 and 3) targeting HDAC6, pooled separately, inhibited BAG3 KD It was found that HDAC3 and HDAC6 provided protection from sarcomere damage in the model and did not affect BAG3 expression. To further confirm which HDACs were the targets of tubulin, acetylated tubulin (Ac-tubulin) levels were also measured in these knockdown studies. It was found that Ac-tubulin levels were significantly higher in HDAC3 and HDAC6 knockdown compared to SCR control.

[0305] HDAC6 inhibition or knockout leads to tubulin hyperacetylation HDAC6 is localized in the cytoplasm (Hubbert et al., 2002; Joshi et al., 2013). In this study, we verified that HDAC6 is predominantly cytoplasmic (~90%) in iPSC-CMs. Using clustered regularly interspaced short palindromic repeats (CRISPR) / Cas9, we isolated an HDAC6 knockout (HDAC6 KO We generated iPSC lines that were successfully differentiated into cardiomyocytes, which showed expression of sarcomeric markers (TNNT2, MYBPC3) and hyperacetylation of tubulin.

[0306] TYA-018 is a highly selective HDAC6 inhibitor Develop a selective HDAC6 inhibitor (TYA-018) and target BAG3 cKO Its efficacy was tested in mouse models. First, biochemical assays were used to ensure high selectivity of TYA-018, measuring its potency against HDAC6 and selectivity against HDAC1 (Figure 7A). As controls, a pan-HDAC inhibitor (Givinostat) and a well-known HDAC6-specific inhibitor (Tubastatin A) were used. In addition, the on-target activity of TYA-018 was measured by measuring Ac-tubulin in iPSC-CMs (Figures 7B-7C). The data suggest that TYA-018 is more potent and selective than Tubastatin A. TYA-018 was further investigated in cell-based assays by measuring acetylated lysines on histones H3 and H4. No off-target activity of TYA-018 against nuclear HDACs was detected, indicating high selectivity (Figure 7D).

[0307] We further confirmed the selectivity of TYA-018 in a full set of biochemical assays using HDAC1-HDAC11 (Figure 8A). We showed that TYA-018 had over 2500-fold selectivity compared to other zinc-dependent HDACs (Figure 8B). In addition, we profiled NAD-dependent HDACs (SIRT1-SIRT6) but observed no activity in the SIRT biochemical assay (data not shown). We used immunostaining to confirm that TYA-018 showed no evidence of off-target HDAC6 activity as measured using acetylated lysine. In the ProBNP assay, we found that TYA-018 did not dose-dependently increase ProBNP levels as seen with divinosta and tubastatin A, indicating that TYA-018 is more selective for HDAC6 than divinosta or tubastatin A (Figure 8C). Finally, the cytotoxicity of TYA-018 in human embryonic kidney cells was investigated, demonstrating a lethal dose (LD50) of 50 μM.

[0308] As a final confirmation, we performed RNA-Seq on WT iPSC-CMs treated with TYA-018, givinostat, and two HDAC6-selective inhibitors (Tubastatin A and Ricorinostat). We showed that increasing selectivity reduced the number of up- and down-regulated transcripts in iPSC-CMs. These findings further confirm the high selectivity of TYA-018 for HDAC6 and ensure that the compound's activity is not associated with transcriptional activation in iPSC-CMs. In addition, treatment of HDAC6 KO cells with TYA-018 does not increase the levels of Ac-tubulin.

[0309] Cardiac-specific knockout of BAG3 in mice leads to heart failure Cardiac-specific BAG3 knockout mice (BAG3 cKO) was used as a model of DCM. As previously reported by Fang and coworkers (2017), this mouse model shows rapid cardiac decline and death due to heart failure. By 5 months of age, mice show a mean ejection fraction (EF) of approximately 30% and a survival rate of approximately 50%. In addition, left ventricular diameter diastolic (LVIDd), left ventricular diameter systolic (LVIDs), and left ventricular mass were significantly decreased in BAG3 mice compared to WT littermates. cKO The BAG3 expression was significantly greater in mice. cKO M-mode echocardiogram tracings from mice showed a rapid decline in cardiac function between 1 and 5 months of age.

[0310] HDAC6 inhibition inhibits BAG3 cKO Prevents progression of heart failure in mice To assess the translatability of findings from the in vitro screen to an in vivo model, a pan-HDAC inhibitor (Givinostat) and an HDAC6-selective inhibitor (Tubastatin A) were used to evaluate BAG3 expression. cKO Efficacy studies were performed in mice. Both inhibitors were used to assess what percentage of efficacy results from inhibition of HDAC6 alone, and whether inhibition of other HDACs would provide additional benefit. In addition, both divinostat and tubastatin A have similar biochemical and cell-based potency for HDAC6 inhibition (Figures 7A-7B).

[0311] When the mice were 1 month old, daily doses of givinostat (30 mg / kg by oral gavage) and tubastatin A (50 mg / kg by intraperitoneal injection) were started (Figure 9A). cKO Mice had significantly reduced cardiac function as measured by EF compared to WT littermate controls (approximately 13%, p<0.0001). Daily administration of both givinostat and tubastatin A prevented the progression of heart failure over the 10-week treatment period (Figures 9B-9E). In addition, LVIDd and LVIDs were significantly reduced in BAG3 mice treated with givinostat (Figures 9F-9G) and tubastatin A (Figures 9H-9I). cKO was significantly decreased in mice.

[0312] Based on these efficacy studies, inhibition of HDAC6 alone may be associated with increased inhibition of BAG3 cKO They concluded that the combination of pan-HDAC inhibitors with pan-HDAC inhibitors was sufficient to provide cardioprotection against heart failure in a mouse model, and polypharmacology associated with pan-HDAC inhibitors did not provide additional cardioprotection in these mice.

[0313] HDAC6 inhibition inhibits BAG3 E455K Protects against heart failure in mice To mimic the patient-specific mutation, we used BAG3 ( E455K We used a second mouse model containing a human mutation in BAG3 (Figure 10A). Mutations in this domain disrupt the interaction with HSP70 and destabilize the chaperone complex that is essential for maintaining protein quality control and homeostasis in cells (Fang et al., 2017). Having found that only HDAC6 inhibition provided sufficient cardioprotection, we also used BAG3 inhibition. E455K A second mouse efficacy study in mice was initiated (Fang et al., 2017). To test whether subsequent intervention could provide protection against heart failure, these mice were treated with Tubastatin A (50 mg / kg IP) at 3 months of age. After 6 weeks of treatment, Tubastatin A reduced BAG3 as measured by EF. E455K Tubastatin A treatment resulted in cardioprotection (Figures 10B-10E) and reduction of LVIDd and LVIDs (Figures 10F-10G) in mice. Most surprisingly, treatment with Tubastatin A resulted in cardioprotection (Figures 10B-10E) and reduction of LVIDd and LVIDs (Figures 10F-10G) in mice. E455K We noticed that male mice had a longer life span (Figures 10H-10I). Protection from premature death due to heart failure was more pronounced in male mice.

[0314] HDAC6 inhibition inhibits BAG3 cKO Prevents heart failure in mice BAG3 cKOThe efficacy of the highly selective HDAC6 inhibitor TYA-018 in mice was tested. In this third efficacy study, mice were treated daily with TYA-018 (15 mg / kg by oral gavage) starting at 2 months of age (FIG. 11A).

[0315] TYA-018 conferred cardioprotection to these mice during the 8-week treatment period, as measured by EF (FIGS. 11B-11C), and significantly reduced LVIDd and LVIDs (FIGS. 11D-6E).

[0316] Because TYA-018 is ultraselective for the HDAC6 isoform (Figures 8A-8C), this efficacy study suggests that HDAC6 inhibition exclusively drives efficacy.

[0317] Processing is BAG3 cKO Well tolerated in mice There was no significant effect on the body weight of mice treated with TYA-018 for 8 weeks. cKO There were no significant differences in platelet counts or neutrophil-to-lymphocyte ratios in mice compared to vehicle controls.The data suggest that daily dosing of TYA-018 was well tolerated in mice.

[0318] Processing is BAG3 cKO Correcting transcriptional profiles in mice A principal components analysis of coding genes in hearts from the three arms of the third efficacy trial with TYA-018 was performed. This analysis identified BAG3 cKOAn overall correction of +TYA-018 coding genes towards their WT littermates was shown (Figure 11H). RNA-Seq data from a selected number of genes are presented as Z-scores in Figure 11I. The data show a correction trend for key sarcomere genes (MYH7, TNNI3, and MYL3) and genes regulating mitochondrial function and metabolism (CYC1, NDUFS8, NDUFB8, PPKARG2). Reductions in inflammatory markers (IL-1β, NLRP3) and apoptotic markers (CASP1, CAPS8) were also shown. RNA-Seq analysis revealed that NPPB expression levels were significantly higher in BAG3 at 4 months of age compared to WT mice. cKO It was shown that TYA-018 treatment increased NPPB levels by approximately four-fold in mice. cKO NPPB expression was reduced by 2-fold in TYA-018-treated BAG3 mice. NPPB expression was inversely correlated with EF (FIG. 11J). QPCR analysis revealed that TYA-018-treated BAG3 mice cKO A significant decrease in NPPB levels in mice was further confirmed to be close to that in WT mice (FIG. 11K).

[0319] By processing, BAG3 cKO Reduced fragmented nuclei and increased LC3 puncta in mice At 4 months of age, after 8 weeks of TYA-018 dosing, hearts were isolated and sectioned from all groups of mice in the third efficacy study. Using immunohistochemistry, we identified BAG3 cKO Significantly larger fragmented nuclei were observed in BAG3 mice compared to WT littermates. TUNEL staining was used to confirm that these fragmented nuclei were apoptotic. TYA-018 treatment reduced the number of fragmented nuclei (p=0.073), which was consistent with the BAG3 expression profile. cKO It was inversely correlated with EF in mice.

[0320] In addition, staining for microtubule-associated protein light chain 3 (LC3) revealed that TYA-018-treated BAGLC3 cKO Larger LC3 puncta and a higher percentage of LC3-positive areas were observed in the hearts of mice. The percentage of LC3-positive areas was significantly higher in BAG3 mice treated with TYA-018 than in mice treated with BAG3. cKOCorrelated with EF in mouse hearts. Western blots from mouse hearts showed that BAG3 cKO In mice, the levels of FLNC, PINK1, and VDAC2 were increased, and the levels of p62 were decreased. cKO These results suggest sarcomere and mitochondrial damage and impaired autophagic flux in mice. TYA-018 treatment increased BAG3 cKO TYA-018 partially restored these markers to WT levels in mice. As expected, TYA-018 treatment significantly increased Ac-tubulin levels in mouse hearts. Tubulin levels were significantly increased by BAG3. cKO HDAC6 expression was significantly increased in mice and levels were unaffected by TYA-018 treatment. These data suggest that inhibition of HDAC6 with TYA-018 protects cardiac function by promoting autophagy and clearing damaged and misfolded proteins, as well as by blocking apoptosis in the heart.

[0321] We next examined five classes of drugs (15 drugs in total) that are either standard of care (SOC) for cardiovascular disease or in late-stage clinical trials for cardiovascular disease to see if they affected Ac-tubulin levels. The data showed that SOC agents had no effect on Ac-tubulin levels or HDAC6 expression in iPSC-CMs, suggesting that cardioprotection from HDAC6 inhibition acts through an independent mechanism not covered by current SOC agents (Figures 12A-12B).

[0322] HDAC6 is increased in the ischemic human heart and in various animal models of DCM Analysis of mouse cardiac biomarkers revealed that HDAC6 protein levels were significantly higher than BAG3 cKOWe found that HDAC6 levels were elevated in mouse hearts compared to WT littermates. We then evaluated cardiac tissue samples from human ischemic heart samples (FIG. 13A) and other mouse models of heart failure (FIG. 13B), including pressure overload using aortic coarctation (TAC), angiotensin II-induced hypertension (Ang II), and myocardial infarction (MI). HDAC6 levels were significantly elevated compared to matched controls. These findings suggest that elevated HDAC6 may be a pathogenic compensatory mechanism in heart failure, and that inhibiting HDAC6 may be protective in familial DCM associated with genes other than BAG3 and heart failure from other causes other than DCM.

[0323] HDAC6 inhibition improves the survival of a second DCM mouse model (MLP KO ) heart failure is prevented HDAC6 inhibition reduces BAG3 KO To demonstrate protection against heart failure beyond the DCM model, a second genetic model (MLP KO We tested the effects of HDAC6 inhibitors in skeletal and cardiac muscle (mice). MLP (or CSRP3) is expressed in cardiac and skeletal muscle and localizes to Z-disks (Arber et al., 1997; Knoll et al., 2010). MLP-deficient mice exhibit sarcomere damage and myofibrillar disorganization and develop dilated cardiomyopathy and heart failure (Arber et al., 1997). In this fourth efficacy study, mice were treated daily with TYA-631 (30 mg / kg by oral gavage) starting at 1.5 months of age (Figure 13SA). TYA-631 is a highly selective HDAC6 inhibitor as measured by cell-based and biochemical assays (Figures S13B-S13D). TYA-631 conferred cardioprotection to these mice over the 9-week dosing period, as shown by EF (Figures S13E-H), and reduced LVIDd and LVIDs (Figures 6F, 6G). TYA-631 is a compound within formula (I), as well as compounds within formulas I(y), Ik, and Ic.

[0324] Example 2 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 for potency against HDAC6 and selectivity against HDAC1 in biochemical assays. 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 3. 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 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9]

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Table 3-76

Table 3-77

[0325] Example 3 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 4. 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 4-1] [Table 4-2]

[0326] The structures, chemical names, and additional biochemical properties of the compounds described in this example are provided below. [Table 13]

Table 14-1

Table 14-2

Table 14-3

Table 14-4

Table 14-5

Table 15-1

Table 15-2

Table 15-3

Table 15-4

Table 16

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[0381] 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 taken 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 composition comprising an HDAC6 inhibitor for use in a method for treating or preventing dilated cardiomyopathy in a subject in need thereof, comprising: The HDAC6 inhibitor is a compound according to formula (I), or a pharma- ceutically acceptable salt thereof; 【Chemistry 113】 R 1 is selected from the group consisting of: 【Table 16】 R a is selected from the group consisting of H, halo, C 1-3 alkyl, cycloalkyl, haloalkyl, and alkoxy; R 2 and R 3 are 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 selected from the group consisting of H, —(SO 2 )R 2 , —(SO 2 )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, C 1 -C 6 alkyl, haloalkyl, cycloalkyl, and heterocyclyl; X 1 is selected from the group consisting of S, O, NH, and NR 6 , where R 6 is selected from the group consisting of C 1 -C 6 alkyl, alkoxy, haloalkyl, cycloalkyl, and heterocyclyl; Y is selected from the group consisting of CR 2 , O, N, S, SO, and SO 2 , and when Y is O, S, SO, or SO 2 , R 5 is absent, and when R 4 and R 5 together with the atom to which they are attached form a cycloalkyl or heterocyclyl, Y is CR 2 or N; A composition wherein n is selected from 0, 1, and 2.

2. The HDAC6 inhibitor is a compound according to formula (Ic), or a pharma- ceutically acceptable salt thereof: 【Chemistry 120】 The composition of claim 1 , wherein R a is H, Me, or F.

3. The composition of claim 1, wherein 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. R 5 is selected from the group consisting of: 【Chemistry 116】 2. The composition of claim 1, wherein R b is selected from the group consisting of halo, alkyl, haloalkyl, alkoxy, haloalkoxy, acyl, sulfonyl, cycloalkyl, heteroaryl, and heterocyclyl; and m is 0, 1, or 2.

5. 2. The composition of claim 1, wherein the HDAC6 inhibitor is selected from the group consisting of: 【Table 18-1】 【Table 18-2】 【Table 18-3】 【Table 18-4】 【Table 18-5】 【Table 18-6】 【Table 18-7】 【Table 18-8】 【Table 18-9】 【Table 18-10】 【Table 18-11】 【Table 18-12】 【Table 18-13】 【Table 18-14】 【Table 18-15】 【Table 18-16】 【Table 18-17】 【Table 18-18】 【Table 18-19】 【Table 18-20】 【Table 18-21】 【Table 18-22】 【Table 18-23】 【Table 18-24】 【Table 18-25】 【Table 18-26】 【Table 18-27】 【Table 18-28】 【Table 18-29】 【Table 18-30】 【Table 18-31】 【Table 18-32】 【Table 18-33】 【Table 18-34】 【Table 18-35】 【Table 18-36】 【Table 18-37】 【Table 18-38】 【Table 18-39】 【Table 18-40】

6. The HDAC6 inhibitor is 【Chemistry 114】 The composition of claim 5, which is (TYA-018) or a pharma- ceutically acceptable salt thereof.

7. The HDAC inhibitor 【Chemistry 117】 or a pharma- ceutically acceptable salt thereof.

8. The HDAC inhibitor, 【Chemistry 118】 or a pharma- ceutically acceptable salt thereof.

9. The HDAC inhibitor 【Chemistry 119】 or a pharma- ceutically acceptable salt thereof.

10. 1. A composition comprising an HDAC6 inhibitor for use in a method for treating or preventing dilated cardiomyopathy in a subject in need thereof, comprising: wherein the HDAC6 inhibitor is a compound according to formula (II): 【Chemistry 115】 n is 0 or 1; X is O, NR ４ , or C.R. ４ R ４’ and Y is a bond, CR ２ R ３ , or S(O) ２ and R １ is selected from the group consisting of H, amido, carbocyclyl, heterocyclyl, aryl, and heteroaryl; R ２ and R ３ are independently H, halogen, alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH ２ )-carbocyclyl, -(CH ２ )-heterocyclyl, -(CH ２ )-aryl, and -(CH ２ )-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’ each independently represents H, an alkyl group, or —CO ２ -Alkyl, carbocyclyl, heterocyclyl, aryl, heteroaryl, -(CH ２ )-carbocyclyl, -(CH ２ )-heterocyclyl, -(CH ２ )-aryl, and -(CH ２ )-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 independently selected from halogen, haloalkyl, oxo, hydroxy, alkoxy, —OCH ３ , -CO ２ CH ３ , -C(O)NH(OH), -CH ３ , morpholine, and -C(O)N-cyclopropyl.

11. 1. A composition comprising an HDAC6 inhibitor for use in a method for treating or preventing dilated cardiomyopathy in a subject in need thereof, comprising: The composition, wherein the HDAC6 inhibitor is CAY10603, tubacin, licorinostat (ACY-1215), citalinostat (ACY-241), ACY-738, QTX-125, CKD-506, nextulastat A, tubastatin A, or HPOB.

12. The composition of claim 1 , wherein the HDAC6 inhibitor is at least 100-fold selective for HDAC6 compared to all other isozymes of HDAC.

13. The composition according to any one of claims 1 to 12, wherein the dilated cardiomyopathy is familial dilated cardiomyopathy.

14. The composition according to any one of claims 1 to 12, wherein the dilated cardiomyopathy is dilated cardiomyopathy caused by one or more BLC2-associated athanogen3 (BAG3) mutations.

15. The composition of any one of claims 1 to 12, wherein the subject has a deleterious mutation in the BAG3 gene.

16. The composition according to any one of claims 1 to 12, wherein the dilated cardiomyopathy is dilated cardiomyopathy caused by one or more muscle LIM protein (MLP) mutations.

17. The composition of any one of claims 1 to 12, wherein the subject has a deleterious mutation in the CSPR3 gene that encodes MLP.

18. The composition of any one of claims 1 to 12, wherein the subject is a human.

19. The composition of any one of claims 1 to 12, wherein administration of the composition restores the ejection fraction of the subject to at least the ejection fraction of a subject not having dilated cardiomyopathy.

20. The composition of any one of claims 1 to 12, characterized in that administration of the composition increases the ejection fraction of the subject compared to the ejection fraction of the subject before treatment.

21. 13. The composition of any one of claims 1 to 12, wherein administration of the composition restores the subject's ejection fraction to at least about 20%, at least about 20%, at least about 30%, at least about 40%, or at least about 50%.

22. 13. The composition of any one of claims 1 to 12, wherein administration of the composition increases the subject's ejection fraction by at least about 5%, at least about 10%, at least about 20%, at least about 30%, or at least about 40%.

23. The composition according to any one of claims 1 to 12, characterized in that administration of said composition reduces HDAC6 activity in the heart of said subject.

24. The composition according to any one of claims 1 to 12, characterized in that administration of the composition prevents heart failure in the subject.

25. The composition according to any one of claims 1 to 12, characterized in that administration of the composition reduces the left ventricular diastolic dimension (LVIDd) of the subject.

26. The composition according to any one of claims 1 to 12, characterized in that administration of the composition reduces the left ventricular systolic diameters (LVIDs) of the subject.

27. The composition according to any one of claims 1 to 12, wherein administration of the composition reduces left ventricular mass in the subject.

28. The composition of any one of claims 1 to 12, wherein the method comprises selecting the HDAC6 inhibitor by performing in vitro testing for selective inhibition of HDAC6 on each member of a plurality of candidate compounds, thereby identifying a selected compound for use as the HDAC6 inhibitor.

29. A pharmaceutical composition for use in a method for treating dilated cardiomyopathy, comprising the HDAC6 inhibitor described in any one of claims 1 to 11.

30. A kit comprising an HDAC6 inhibitor according to any one of claims 1 to 11 and instructions for use in a method for treating dilated cardiomyopathy.

31. A method for identifying a compound for the treatment of dilated cardiomyopathy, comprising contacting a cell culture comprising cells having an inactivating mutation in BAG3 with each member of a plurality of candidate compounds, and selecting a compound that reduces sarcomere damage in the cells.

32. A composition comprising a selected compound for use in a method of treating dilated cardiomyopathy in a subject in need thereof, said method comprising: a) contacting a cell culture comprising cells having an inactivating mutation in BAG3 with each member of a plurality of candidate compounds to identify compounds and selecting said selected compounds as reducing sarcomere damage; b) administering said selected compound to said subject.

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