Compounds, compositions, and methods for treating metabolic disorders

Abstract

The present disclosure provides compounds of Formula (I) or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and methods for treating metabolic diseases and disorders with a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, wherein the disease or disorder is obesity, cirrhosis, myocardial infraction, or alcoholism. (I), wherein L, R2, and n are defined herein.

Description

COMPOUNDS, COMPOSITIONS, AND METHODS FOR TREATINGMETABOLIC DISORDERSBACKGROUND

[0001] Taurine is a conditionally essential micronutrient and very abundant amino sulfonic acid that is found in mammalian tissues and many foods29. Levels of taurine are especially high in excitable tissues such as the heart, eyes, brain, and muscles5. Taurine has been described to have pleiotropic cellular and physiologic functions, particularly in the context of mitochondrial function and whole-body metabolism9 11. Genetic reduction of tissue taurine levels leads to muscle atrophy12 13, decreased exercise capacity14, and mitochondrial dysfunction in multiple tissues12 15. Conversely, taurine supplementation has been reported to reduce mitochondrial redox stress9, enhance exercise performance16and suppress body weight17.

[0002] The biochemistry and enzymology of taurine metabolism has attracted considerable research interest. In the endogenous taurine biosynthesis pathway, cysteine is metabolized via cysteine dioxygenase (CDO) and cysteine sulfinic acid decarboxylase (CSAD) to generate hypotaurine18 19, which is subsequently oxidized by flavin-containing monooxygenase 1 (FMO1) to produce taurine20. In addition, cysteine can undergo an alternative pathway via cysteamine and 2-aminoethanethiol dioxygenase (ADO)21. Downstream of taurine itself are several taurine-containing derivatives that include taurocholate, taurocyamine, taurocyamine phosphate, and N- acety Itaurine4. The only enzyme known to catalyze one of these downstream pathways is bile acid-coenzyme A:amino acid N-acyltransferase (BAAT), which conjugates taurine to bile acyl-CoAs to produce taurocholate and other bile salts22. Beyond BAAT as the sole example, the molecular identities of additional enzymes that mediate the metabolism of taurine to taurine-containing metabolites have not yet been established.

[0003] The biochemical interconversion of taurine and N-acetyltaurine is of particular interest for several reasons. First, N-acetyltaurine is an abundant endogenous metabolite whose levels are dynamically regulated by diverse physiologic perturbations that increase taurine and / or acetate flux, including endurance exercise7’17, alcohol consumption623, and nutritional taurine supplementation17. Second, N-acetyltaurine exhibits chemical structural similarities with signaling molecules including the neurotransmitter acetylcholine24and the glucoregulatory long-chain N-fatty acyl taurines25. Third, a taurine N-acetyltransferase biochemical activity has been detected in cells6’7, demonstrating that the production of N-acetyltaurine is not simply a byproduct of taurine metabolism, but rather an enzymatically regulated biochemical transformation.

[0004] The orphan body mass index-associated enzyme PTER (phosphotriesterase-related) is the principal mammalian taurine N-acetyltransferase / hydrolase26. In vitro, recombinant PTER catalyzes bidirectional taurine N-acetylation with free acetate as well as the reverse N-acetyltaurine hydrolysis reaction. Genetic ablation of PTER in mice results in complete loss of tissue taurine N-acetyltransferase / hydrolysis activities and systemic elevation of N-acetyltaurine levels. Upon stimuli that increase taurine levels, PTER knockout mice exhibit lower body weight, reduced adiposity, and improved glucose homeostasis. These phenotypes are recapitulated by administration of N-acetyltaurine to wild-type mice. Lastly, the anorexigenic and anti-obesity effects of N-acetyltaurine require functional GDNF family receptor a-like (GFRAL) receptors. Together, these data uncover enzymatic control of a previously enigmatic pathway of secondary taurine metabolism linked to energy balance. Thus, there is a need for PTER inhibitors which can be useful as therapeuticsSUMMARY

[0005] Provided herein are compounds and pharmaceutical compositions, and their use in treating diseases and disorders associated with aberrant PTER activity, such as obesity, cirrhosis, myocardial infraction, or alcoholism.

[0006] In some aspects, provided herein are compounds of Formula (I), whereinor a pharmaceutically acceptable salt thereof; wherein:L is Cs galkylene or Ca-galkenylene, wherein the alkylene or alkenylene backbone is interrupted with 1 , 2, or 3 heteroatoms independently selected from NR1, O, and S; andR1is H or CH3; each R2is independently N(RN)2 or NO2; each RNis independently H or Ci-salkyl; and n Is O, 1 , or 2.

[0007] In other aspects, the disclosure provides pharmaceutical formulations comprising a compound or salt of Formula (I) and a pharmaceutically acceptable excipient.

[0008] In still other aspects, the disclosure provides methods of inhibiting PTER activity in a cell comprising contacting the cell with a compound or salt of Formula (I) in an amount effective to inhibit PTER activity.

[0009] In yet other aspects, the disclosure provides methods of treating a disease or disorder associated with aberrant PTER activity in a subject, comprising administering to the subject a therapeutically effective amount of a compound or salt of Formula (I).

[0010] In yet other aspects, the disclosure provides methods for treating weight gain associated with discontinuation of a GLP-1 R agonist in a subject, comprising administering to the subject a therapeutically effective amount of a compound or salt of Formula (I).BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 shows the PTER and HDAC inhibition activity of known HDAC inhibitors vorinostat, droxinostat, and scriptaid.

[0012] Figure 2 shows (a) the HDAC and PTER inhibitory activity for compounds 32-42, as disclosed herein, and (b) for Compound 41 alone, showing Compound 41 has a greater than 250 x selectivity for inhibition of PTER compared to HDAC.

[0013] Figures 3 and 4 show the inhibitory activity of Compound 41 for HDACs 1-11 and SIRTs 1-4, respectively.

[0014] Figure 5 shows that Compound 41 increases N-acetyltaurine levels in lean and obese mice.

[0015] Figure 6 shows that administration of Compound 41 to obese mice decreases food intake over 24- hours post dose.

[0016] Figure 7 shows the relative changes in body weight for mice injected with either vehicle or compound 41 relative to cumulative food intake.

[0017] Figure 8 shows the cumulative food intake for mice injected with either vehicle or compound 41 over 24 hours.

[0018] Figure 9 shows the respiratory exchange ratio for mice injected with either vehicle or compound 41 in light and dark.

[0019] Figure 10 shows the activity of PTER for mice injected with either vehicle or compound 41 in light and dark.

[0020] Figure 11 shows the cumulative food intake for mice injected with either vehicle or compound 41 over 27 days.

[0021] Figure 12 shows the relative changes in body weight for mice injected with either vehicle or compound 41 over 27 days.

[0022] Figure 13 shows the relative changes in body weight for mice injected with either vehicle, semaglutide, or semaglutide and compound 41 over 7 days.

[0023] Figure 14 shows the cumulative food intake for mice injected with either vehicle, semaglutide, or semaglutide and compound 41 over 7 days.

[0024] Figure 15 shows the relative changes in body weight for mice injected with either vehicle, semaglutide alone, semaglutide with vehicle, or semaglutide and compound 41 over 25 days.

[0025] Figure 16 shows compound 41 and PTERamide.

[0026] Figure 17 shows the relative activity of compound 41 and PTERamide.

[0027] Figure 18 shows the cumulative food intake for mice injected with either vehicle, or PTERamide over 5 days.

[0028] Figure 19 shows the relative changes in body weight for mice injected with either vehicle or PTERamide over 5 days.DETAILED DESCRIPTION

[0029] In general, the disclosure relates to compounds or pharmaceutically acceptable salts of Formula (I) that are useful as PTER inhibitors, pharmaceutical formulations comprising a compound or pharmaceutically acceptable salt of Formula (I), methods of inhibiting PTER activity in a cell comprising contacting the cell with a compound or salt of Formula (I) in an amount effective to inhibit PTER activity, and methods of treating a disease or disorder associated with aberrant PTER activity in a subject, comprising administering to the subject a therapeutically effective amount of a compound or salt of Formula (I).

[0030] Provided herein are compounds having a structure of Formula (I):pharmaceutically acceptable salt thereof; wherein: L is Cs-galkylene or Cs-galkenylene, wherein the alkylene or alkenylene backbone is interrupted with 1 , 2, or 3 heteroatoms independently selected from NR1, O, and S; R1is H or CH3; each R2is independently N(RN)g or NO2; each RNis independently H or C 1.3a! ky I ; and n is 0, 1 , or 2.

[0031] In some cases, L is 6-10 atoms long. In various cases L is 8 atoms long.

[0032] In some cases, L is an alkylene and the alkylene backbone is interrupted with 1 heteroatom. In some cases, L is an alkenylene and the alkenylene backbone is interrupted with 1 heteroatom. In various cases, the heteroatom is O. In some cases, the heteroatom is NR1. In some cases, R1is methyl. In some cases, R1is H.

[0033] In some cases, L is an alkylene and the alkylene backbone is interrupted with 2 heteroatoms. In some cases, L is an alkenylene and the alkenylene backbone is interrupted with 2 heteroatoms. In various cases, at least one heteroatom is O. In various cases, each heteroatom is 0. In some cases, at least one heteroatom is NR1. In some cases, each heteroatom is NR1. In some cases, one heteroatom is 0 and the other heteroatom is NR1. In some cases, each R1is methyl. In some cases, each R1is H.

[0034] In some cases, L is an alkylene and the alkylene backbone is interrupted with 3 heteroatoms. In some cases, L is an alkenylene and the alkenylene backbone is interrupted with 3 heteroatoms. In various cases, at least one heteroatom is 0. In various cases, each heteroatom is 0. In some cases, at least one heteroatom is NR1. In some cases, each heteroatom is NR1. In some cases, two heteroatoms are 0 and the other heteroatom is NR1. In some cases, one heteroatom is 0 and the other heteroatoms are NR1. In some cases, each R1is methyl.

[0035] In cases where there are two or three heteroatoms, in some embodiments, at least one heteroatom is S.

[0036] In various cases, the compound or salt is selected from

[0037] In some cases, the compound or salt thereof has a structure as recited in Table A.TABLE A

[0038] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the relevant art. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, representative illustrative methods and materials are described herein.

[0039] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0040] It is noted that, as used herein and in the appended claims, the singular forms "a”, "an”, and "the” include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely,” "only” and the like in connection with the recitation of claim elements, or use of a "negative” limitation.

[0041] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present invention. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.Definitions

[0042] The term "aberrant," as used herein, refers to different from normal. When used to describe enzymatic activity, aberrant refers to activity that is greater or less than a normal control or the average of normal nondiseased control samples. Aberrant activity may refer to an amount of activity that results in a disease, where returning the aberrant activity to a normal or non-disease-associated amount (e.g., by administering a compound or using a method as described herein), results in reduction of the disease or one or more disease symptoms. The aberrant activity can be measured by measuring the modification of a substrate of the enzyme in question; a difference of greater or equal to a 2-fold change in activity could be considered as aberrant. Aberrant activity could also refer to an increased dependence on a particular signaling pathway as a result of a deficiency in a separate complementary pathway.

[0043] The term "alkylene,” as used herein, refers to a divalent alkyl group. As used herein, the term "alkyl” refers to straight chained (not branched) saturated hydrocarbon groups containing one to thirty carbon atoms, for example, one to twenty carbon atoms, or one to ten carbon atoms. The term Cnmeans the alkyl group has “n” carbon atoms. For example, C4 alkyl refers to an alkyl group that has 4 carbon atoms. Ci-Ce alkyl refers to an alkyl group having a number of carbon atoms encompassing the entire range (e.g., 3 to 9 carbon atoms), as well as all subgroups (e.g., 3-9, 3-8, 4-9, 3-7, 3, 4, 5, 6, 7, 8, and 9 carbon atoms). Nonlimiting examples of alkyl groups include, n-propyl, n-butyl, and n-pentyl. Unless otherwise indicated, an alkylene group can be an unsubstituted alkylene group or a substituted alkylene group. An optionally substituted alkylene is an alkylene that is optionally substituted as described herein for alkyl. The alkylene moiety as used herein is interrupted with 1 , 2, or 3 heteroatoms, meaning that the alkylene chain has in the backbone 1-3 heteroatoms (e.g., Cxalkylene- Heteroatom-Cxalkylene-[Heteroatom]o-i-Cxalkylene-[Heteroatom]o-i-Cxalkylene, where all the xs add up to 3-9 for the alkylene chain). When 2 or 3 heteroatoms are present, two heteroatoms can be in sequence (i.e., Heteroatom 1-Heteroatom2, without a carbon atom in between), but more preferably are not in sequence.

[0044] The term "alkenylene, ” as used herein, refers to a divalent alkenyl group. As used herein, the term"alkenyl” refers to straight chained (not branched) unsaturated hydrocarbon groups containing two to thirty carbon atoms, for example, two to twenty carbon atoms, or two to ten carbon atoms. The term Cnmeans the alkenyl group has “n” carbon atoms. For example, C4 alkenyl refers to an alkenyl group that has 4 carbon atoms. C2-C6 alkenyl refers to an alkenyl group having a number of carbon atoms encompassing the entire range (e.g., 2 to 6 carbon atoms), as well as all subgroups (e.g., 2-5, 2-4, 2-3, 3-6, 3-5, 3-4, 4-6, 4-5, 5-6, 2, 3, 4, 5, and 6 carbon atoms). Nonlimiting examples of alkenyl groups include ethylene and propylene. Unless otherwise indicated, an alkenylene group can be an unsubstituted alkenylene group or a substituted alkenylene group. An optionally substituted alkenylene is an alkenylene that is optionally substituted as described herein for alkylene. The alkenylene moiety as used herein is interrupted with 1 , 2, or 3 heteroatoms, meaning that the alkenylene chain has in the backbone 1-3 heteroatoms (e.g., (e.g., Cxalkenylene-Heteroatom-Cxalkenylene-[Heteroatom]o-i- Cxalkenylene-[Heteroatom]o-i-Cxalkenylene, where all the xs add up to 3-9 for the alkenylene chain). When 2 or3 heteroatoms are present, two heteroatoms can be in sequence (i.e., Heteroatom1-Heteroatom2, without a carbon atom in between), but more preferably are not in sequence.

[0045] As used herein, reference to an element, whether by description or chemical structure, encompasses all isotopes of that element unless otherwise described. By way of example, the term "hydrogen” or “H” in a chemical structure as used herein is understood to encompass, for example, not only1H, but also deuterium (2H), tritium (3H), and mixtures thereof unless otherwise denoted by use of a specific isotope. Other specific nonlimiting examples of elements for which isotopes are encompassed include carbon, phosphorous, idodine, and fluorine.

[0046] The terms "pharmaceutical composition” and "pharmaceutical formulation” are interchangeable and, as used herein, represent a composition containing a compound described herein, formulated with a pharmaceutically acceptable excipient, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other formulation described herein.

[0047] The term "pharmaceutically acceptable excipient” or "pharmaceutically acceptable carrier,” as used interchangeably herein, refers to any ingredient other than the compounds described herein (e.g., a vehicle capable of suspending or dissolving the active compound) and having the properties of being nontoxic and noninflammatory in a patient. Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners, or waters of hydration. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0048] The term "pharmaceutically acceptable salt,” as use herein, represents those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and in Pharmaceutical Salts: Properties,Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting the free base group with a suitable organic acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydrogen sulfate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like.

[0049] The term "therapeutically effective amount,” as used herein, means the amount of a compound or a pharmaceutically acceptable salt thereof that is sufficient to treat obesity, cirrhosis, myocardial infarction, or alcoholism.

[0050] The term "subject,” as used herein, represents a human or non-human animal (e.g., a mammal) that is suffering from, or is at risk of, disease or condition, as determined by a qualified professional (e.g., a doctor or a nurse practitioner) with or without known in the art laboratory test(s) of sample(s) from the subject. Preferably, the subject is a human. Non-limiting examples of diseases and conditions include diseases such as, for example, obesity, cirrhosis, myocardial infarction, or alcoholism.

[0051] "Treatment” and "treating," as used herein, refer to the medical management of a subject with the intent to improve, ameliorate, stabilize, prevent, or cure a disease or condition. This term includes active treatment (treatment directed to improve the disease or condition); causal treatment (treatment directed to the cause of the associated disease or condition); palliative treatment (treatment designed for the relief of symptoms of the disease or condition); preventative treatment (treatment directed to minimizing or partially or completely inhibiting the development of the associated disease or condition); and supportive treatment (treatment employed to supplement another therapy). A disease or condition may be, for example, obesity, cirrhosis, myocardial infarction, or alcoholism.Embodiments of the Disclosure

[0052] In embodiment 1 , the present disclosure provides a compound having a structure of Formula (I):or a pharmaceutically acceptable salt thereof; wherein:L is Cs galkylene or Ca-galkenylene, wherein the alkylene or alkenylene backbone is interrupted with 1 , 2, or 3 heteroatoms independently selected from NR1, 0, and S; andR1is H or CH3; each R2is independently N(RN)2 or NO2; each RNis independently H or Ci-salkyl; and n Is O, 1 , or 2.

[0053] Embodiment 2 provides a compound or salt of embodiment 1, wherein L is 6-10 atoms long.

[0054] Embodiment 3 provides a compound or salt of embodiment 2, wherein L is 8 atoms long.

[0055] Embodiment 4 provides a compound or salt of any one of embodiments 1 to 3, wherein the alkylene backbone is interrupted with 1 heteroatom.

[0056] Embodiment 5 provides a compound or salt of embodiment 4, wherein the heteroatom is O.

[0057] Embodiment 6 provides a compound or salt of embodiment 4, wherein the heteroatom is NR1.

[0058] Embodiment 7 provides a compound or salt of any one of embodiments 1 to 3, wherein the alkylene backbone is interrupted with 2 heteroatoms.

[0059] Embodiment 8 provides a compound or salt of embodiment 7, wherein at least one heteroatom is O.

[0060] Embodiment 9 provides a compound or salt of embodiment 8, wherein each heteroatom is O.

[0061] Embodiment 10 provides a compound or salt of embodiment 7 or 8, wherein at least one heteroatom is NR1.

[0062] Embodiment 11 provides a compound or salt of embodiment 10, wherein each heteroatom is NR1.

[0063] Embodiment 12 provides a compound or salt of embodiment 10, wherein one heteroatom is O and the other heteroatom is NR1.

[0064] Embodiment 13 provides a compound or salt of any one of embodiments 1 to 3, wherein the alkylene backbone is interrupted with 3 heteroatoms.

[0065] Embodiment 14 provides a compound or salt of embodiment 13, wherein at least one heteroatom is O.

[0066] Embodiment 15 provides a compound or salt of embodiment 14, wherein each heteroatom is O.

[0067] Embodiment 16 provides a compound or salt of embodiment 13 or 14, wherein at least one heteroatom is NR1.

[0068] Embodiment 17 provides a compound or salt of embodiment 16, wherein each heteroatom is NR1.

[0069] Embodiment 18 provides a compound or salt of embodiment 13, wherein at least one heteroatom is O and at least one heteroatom is NR1.

[0070] Embodiment 19 provides a compound or salt of any one of embodiments 1 to 4, 7, 8, 10, 13, 14, 16, and 18, wherein at least one heteroatom is S.

[0071] Embodiment 20 provides a compound or salt of any one of embodiments 1 to 4, 6 to 8, 10 to 14, and 16 to 19, wherein R1is CH3.

[0072] Embodiment 21 provides a compound or salt of any one of embodiments 1 to 4, 6 to 8, 10 to 14, and 16 to 19, wherein R1is H.

[0073] Embodiment 22 provides a compound or salt of embodiment 1 selected from

[0074] Embodiment 23 provides a compound or salt thereof having a structure according to Table A.

[0075] Embodiment 24 provides a pharmaceutical formulation comprising the compound or salt of any one of embodiments 1 to 23 and a pharmaceutically acceptable excipient.

[0076] Embodiment 25 provides a method of inhibiting PTER activity in a cell comprising contacting the cell with the compound or salt of any one of embodiments 1 to 23 in an amount effective to inhibit PTER activity.

[0077] Embodiment 26 provides a method of treating a disease or disorder associated with aberrant PTER activity in a subject, comprising administering to the subject a therapeutically effective amount of the compound or salt of any one of embodiments 1 to 23.

[0078] Embodiment 26 provides a method of embodiment 25, wherein the disease or disorder is obesity, cirrhosis, myocardial infarction, or alcoholism.

[0079] Embodiment 27 provides a method of embodiments 24 or 25, further comprising administering an additional therapeutic agent.

[0080] Embodiment 28 provides a method of embodiment 27, wherein the additional therapeutic agent is a GLP-1R agonist.

[0081] Embodiment 29 provides a method of treating weight gain associated with discontinuation of a GLP-1R agonist in a subject, comprising administering to the subject a therapeutically effective amount of the compound or salt of any one of embodiments 1 to 21 .

[0082] Embodiment 30 provides a method of embodiment 28 or 29, wherein the GLP-1 R agonist is semaglutide.Pharmaceutically Acceptable Salts

[0083] The disclosure also includes pharmaceutically acceptable salts of the compounds, and pharmaceutical compositions including the compounds as disclosed herein and a pharmaceutically acceptable carrier. The disclosure provides pharmaceutically acceptable salts of various compounds disclosed herein.Pharmaceutical Compositions

[0084] The compounds can be formulated into pharmaceutical formulation for administration to human subjects in a biologically compatible form suitable for administration in vivo. Pharmaceutical compositions typically include a compound as described herein and a pharmaceutically acceptable excipient.

[0085] The compounds described herein can also be used in the form of the free base, in the form of salts, zwitterions, solvates, or pharmaceutical compositions thereof. All forms are within the scope of the disclosure. The compounds, salts, zwitterions, solvates, or pharmaceutical compositions thereof, may be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. The compounds used in the methods described herein may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, or transdermal administration, and the pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

[0086] For human use, a compound of the disclosure can be administered alone or in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceuticalpractice. Pharmaceutical compositions for use in accordance with the present disclosure thus can be formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries that facilitate processing of a compound of the disclosure into preparations which can be used pharmaceutically.

[0087] This disclosure also includes pharmaceutical compositions which can contain one or more pharmaceutically acceptable carriers. In making the pharmaceutical compositions of the disclosure, the active ingredient is typically mixed with an excipient, diluted by an excipient, or enclosed within such a carrier in the form of, for example, a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it can be a solid, semisolid, or liquid material (e.g., normal saline), which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, and soft and hard gelatin capsules. As is known in the art, the type of diluent can vary depending upon the intended route of administration. The resulting compositions can include additional agents, e.g., preservatives.

[0088] The excipient or carrier is selected on the basis of the mode and route of administration. Suitable pharmaceutical carriers, as well as pharmaceutical necessities for use in pharmaceutical formulations, are described in Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005), a well-known reference text in this field, and in the USP / NF (United States Pharmacopeia and the National Formulary). Examples of suitable excipients are lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. The formulations can additionally include: lubricating agents, e.g., talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents, e.g., methyl- and propylhydroxy-benzoates; sweetening agents; and flavoring agents. Other exemplary excipients are described in Handbook of Pharmaceutical Excipients, 6th Edition, Rowe et al., Eds., Pharmaceutical Press (2009).

[0089] These pharmaceutical compositions can be manufactured in a conventional manner, e.g., by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes. Methods well known in the art for making formulations are found, for example, in Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005), and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York. Proper formulation is dependent upon the route of administration chosen. The formulation and preparation of such compositions is well-known to those skilled in the art of pharmaceutical formulation. In preparing a formulation, the active compound can be milled to provide the appropriate particle size prior to combining with the other ingredients. If the active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh. If the active compound is substantially water soluble, the particle size can be adjusted by milling to provide a substantially uniform distribution in the formulation, e.g., 40 mesh.Dosages

[0090] The dosage of the compound used can vary depending on many factors, e.g., the pharmacodynamic properties of the compound; the mode of administration; the age, health, and weight of the recipient; the nature and extent of the symptoms; the frequency of the treatment, and the type of concurrent treatment, if any; and the clearance rate of the compound in the animal to be treated. One of skill in the art can determine the appropriate dosage based on the above factors. The compounds may be administered initially in a suitable dosage that may be adjusted as required, depending on the clinical response. In general, a suitable daily dose of a compound of the disclosure will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.Routes of Administration

[0091] Exemplary routes of administration of the compounds (e.g., a compound of the disclosure), or pharmaceutical compositions thereof, used in the present disclosure include oral, sublingual, buccal, transdermal, intradermal, intramuscular, parenteral, intravenous, intra-arterial, intracranial, subcutaneous, intraorbital, intraventricular, intraspinal, intraperitoneal, intranasal, inhalation, and topical administration. The compounds desirably are administered with a pharmaceutically acceptable carrier. Pharmaceutical compositions of the compounds described herein formulated for treatment of the disorders described herein are also part of the present disclosure. Oral administration is a preferred route of administration in the methods of the disclosure.

[0092] Compositions for Oral Administration: The pharmaceutical compositions contemplated by the disclosure include those formulated for oral administration ("oral dosage forms”). Oral dosage forms can be, for example, in the form of tablets, capsules, a liquid solution or suspension, a powder, or liquid or solid crystals, which contain the active ingredient(s) in a mixture with non-toxic pharmaceutically acceptable excipients. These excipients may be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starches including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulating and disintegrating agents (e.g., cellulose derivatives including microcrystalline cellulose, starches including potato starch, croscarmellose sodium, alginates, or alginic acid); binding agents (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, carboxymethylcellulose sodium, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); and lubricating agents, glidants, and antiadhesives (e.g., magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenated vegetable oils, or talc). Other pharmaceutically acceptable excipients can be colorants, flavoring agents, plasticizers, humectants, buffering agents, and the like.

[0093] Compositions for oral administration may also be presented as chewable tablets, as hard gelatin capsules where the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin), or as soft gelatin capsules where the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin, or olive oil.Powders, granulates, and pellets may be prepared using the ingredients mentioned above under tablets and capsules in a conventional manner using, e.g., a mixer, a fluid bed apparatus or a spray drying equipment.

[0094] Controlled release compositions for oral use may be constructed to release the active drug by controlling the dissolution and / or the diffusion of the active drug substance. Any of a number of strategies can be pursued in order to obtain controlled release and the targeted plasma concentration versus time profile. In one example, controlled release is obtained by appropriate selection of various composition parameters and ingredients, including, e.g., various types of controlled release compositions and coatings. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles, patches, and liposomes. In certain embodiments, compositions include biodegradable, pH, and / or temperature-sensitive polymer coatings.

[0095] Dissolution- or diffusion- controlled release can be achieved by appropriate coating of a tablet, capsule, pellet, or granulate composition of compounds, or by incorporating the compound into an appropriate matrix. A controlled release coating may include one or more of the coating substances mentioned above and / or, e.g., shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitostearate, ethylcellulose, acrylic resins, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinyl pyrrolidone, polyethylene, polymethacrylate, methylmethacrylate, 2- hydroxymethacrylate, methacrylate hydrogels, 1,3 butylene glycol, ethylene glycol methacrylate, and / or polyethylene glycols. In a controlled release matrix formulation, the matrix material may also include, e.g., hydrated methylcellulose, carnauba wax and stearyl alcohol, carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and / or halogenated fluorocarbon.

[0096] The liquid forms in which the compounds and compositions of the present disclosure can be incorporated for administration orally include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, and flavored emulsions with edible oils, e.g., cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0097] Compositions for Parenteral Administration: The compounds described herein for use in the methods of the disclosure can be administered in a pharmaceutically acceptable parenteral (e.g., intravenous or intramuscular) composition as described herein. The pharmaceutical composition may also be administered parenterally (intravenous, intramuscular, subcutaneous or the like) in dosage forms or compositions containing conventional, non-toxic pharmaceutically acceptable carriers and adjuvants. In particular, compositions suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the composition isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. For example, to prepare such a composition, the compounds of the disclosure may be dissolved or suspended in a parenterally acceptable liquid vehicle. Among acceptable vehicles and solvents that may be employed are water, water adjusted to a suitable pH by addition of an appropriate amount of hydrochloricacid, sodium hydroxide or a suitable buffer, 1 ,3-butanediol, Ringer's solution and isotonic sodium chloride solution. The aqueous composition may also contain one or more preservatives, for example, methyl, ethyl, or n- propyl p-hydroxybenzoate. Additional information regarding parenteral compositions can be found, for example, in the United States Pharmacopeia-National Formulary (USP-NF), herein incorporated by reference.

[0098] The parenteral composition can be any of the five general types of preparations identified by the USP- NF as suitable for parenteral administration:

[0099] (1) "Drug Injection:” a liquid preparation that is a drug substance (e.g., a compound of the disclosure), or a solution thereof;

[0100] (2) "Drug for Injection:” the drug substance (e.g., a compound of the disclosure) as a dry solid that will be combined with the appropriate sterile vehicle for parenteral administration as a drug injection;

[0101] (3) "Drug Injectable Emulsion:” a liquid preparation of the drug substance (e.g., a compound of the disclosure) that is dissolved or dispersed in a suitable emulsion medium;

[0102] (4) "Drug Injectable Suspension:” a liquid preparation of the drug substance (e.g., a compound of the disclosure) suspended in a suitable liquid medium; and

[0103] (5) "Drug for Injectable Suspension:” the drug substance (e.g., a compound of the disclosure) as a dry solid that will be combined with the appropriate sterile vehicle for parenteral administration as a drug injectable suspension.

[0104] Exemplary compositions for parenteral administration include solutions of the compound prepared in water suitably mixed with a surfactant, e.g., hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable compositions are described, for example, in Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005) and in The United States Pharmacopeia: The National Formulary (USP 36 NF31), published in 2013.

[0105] Compositions for parenteral administration may, for example, contain excipients, sterile water, or saline, polyalkylene glycols, e.g., polyethylene glycol, oils of vegetable origin, or hydrogenated napthalenes. Biocompatible, biodegradable lactide polymer, lactide / glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be used to control the release of the compounds. Other potentially useful parenteral delivery systems for compounds include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. Compositions for inhalation may contain excipients, for example, lactose, or may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, glycocholate and deoxycholate, or may be oily solutions for administration in the form of nasal drops, or as a gel.

[0106] The parenteral composition can be formulated for prompt release or for sustained / extended release of the compound. Exemplary compositions for parenteral release of the compound include: aqueous solutions, powders for reconstitution, cosolvent solutions, oil / water emulsions, suspensions, oil-based solutions, liposomes, microspheres, and polymeric gels.Methods of Use

[0107] The compounds disclosed herein are useful as PTER inhibitors. Blocking PTER increases N- acetyltaurine, which can lead to a number of pharmaceutically beneficial results in the treatment of certain cardiometabolic disorders such as, for example, cardiovascular disease, chronic kidney disease, diabetes, insulin resistance, obesity, and non-alcoholic liver disease, wherein the cardiovascular disease may be a myocardial infarction heart disease, heart attack, heart failure, stroke, or vascular disease, and wherein the non-alcoholic liver disease may be non-alcoholic fatty liver disease, cirrhosis, or non-alcoholic steatohepatitis.

[0108] In various embodiments, the compounds disclosed herein are selective for PTER compared to other deacetylases, e.g., histone deacetylase (HDAC), e.g., one or more of HDAC1, HDAC2, HDAC3, HDAC4, HDAC5, HDAC6, HDAC7, HDAC8, HDAC9, HDAC10, and HDAC11, and / or stress responsive protein deacetylase, sirtuin, e.g., one or more of SIRT1 , SIRT2, SIRT3, and SIRT4. In various cases, the compound inhibits PTER 2x more than it inhibits an HDAC or SIRT (as measured by IC50 data), e.g., at least 3x more selective for PTER, at least 4 x, at least 5 x, at least 6x, at least 7x, at least 8x, at least 9x, at least 10x, at least 15x, at least 20x, at least 25x, at least 30x, at least 40x, at least 50x, at least 60x, at least 75x, at least 10Ox, at least 125x, at least 150x, at least 175x, or at least 200x or more selective for PTER compared to an HDAC or SIRT.

[0109] Inhibition of PTER, HDAC, and SIRT can be assessed by any means available, and in some cases is assessed in a manner as described in the examples below.

[0110] In various embodiments, the compounds disclosed herein can be administered in conjunction with, or combined with, an additional therapeutic agent, including but not limited to, GLP-1R agonists. GLP-1R agonists are known in the art. Some contemplated GLP-1R agonists include exenatide, exenatide extended-release, dulaglutide, liraglutide, lixisenatide, semaglutide, tirzepatide, cotadutide, noiiglutide, oxyntomodulin (e.g., mazdutide), retatrutide, albiglutide, beinaglutide and PEG-loxenatide, pemvidutide, and danuglipron. In various embodiments, the GLP-1R agonist is semaglutide.

[0111] Also provided herein are methods for treating weight gain associated with discontinuation of a GLP-1R agonist in a subject, comprising administering to the subject a therapeutically effective amount of a compound or salt disclosed herein. Generally, the GLP-1R agonist can be any GLP-1R agonist known in art. In various embodiments, the GLP-1R agonist is semaglutide.

[0112] The following examples are meant to illustrate the disclosure. They are not meant to limit the disclosure in any way.EXAMPLESCompound Synthesis

[0113] Starting materials, reagents, and solvents were purchased from commercial suppliers and were used without further purification unless otherwise noted. All reactions were monitored using a Waters Acquity UPLC / MS system (Waters PDA eA Detector, QDa Detector, Sample manager - FL, Binary Solvent Manager) using Acquity UPLC® BEH C18 column (2.1 x 50 mm, 1.7 pm particle size): solvent gradient = 85% A at 0 min, 1 % A at 1 .7 min; solvent A = 0.1 % formic acid in Water; solvent B = 0.1 % formic acid in Acetonitrile; flow rate : 0.6 mL / min. Reaction products were purified by flash column chromatography using Combi Fl ash®Rf with Teledyne Isco RediSep® normal-phase silica flash columns (4 g, 12 g, 24 g, 40 g or 80 g) and Waters HPLC system using SunFireTM Prep C18 column (19 x 100 mm, 5 pm particle size): solvent gradient = 80% A at 0 min, 10% A at 25 min; solvent A = 0.035% TFA in Water; solvent B = 0.035% TFA in MeOH; flow rate : 25 mL / min.1H NMR spectra were recorded on 500 MHz Bruker Avance III spectrometers. Chemical shifts are reported in parts per million (ppm, 5) downfield from tetramethylsilane (TMS). Coupling constants (J) are reported in Hz. Spin multiplicities are described as br (broad), s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet). Purities of assayed compounds were in all cases greater than 95%, as determined by reverse-phase LC-Ms analysis.Compound 41

[0114] Compound 41 was prepared using the following reactions.

[0115] Step 1:

[0116] To a solution of 1 H,3H-benzo[de]isochromene-1, 3-dione (1000 mg, 1.0 Eq, 5.046 mmol) in DMF (5 mL) was added tert-butyl 3-(2-(2-aminoethoxy)ethoxy)propanoate (1.236 g, 1.05 Eq, 5.298 mmol) . Then the mixture was stirred at 125 °C for 8 hour. The reaction mixture was concentrated by removing the solvents, and purified with CombiFlash chromatography using methanol with 0 to 10% methylene chloride as eluent to provide Intermediate [III] (1.3g, 62%). MS (ESI): m / z = 436.0 [M+Na]+.

[0118] To a solution of Intermediate [III] (1 g, 1 Eq, 2 mmol) in dichloromethane (6 mL) was added TFA (3 mL) and the mixture was stirred at 25 °C for 1 hour. Then the solvents were removed, to provide crude Intermediate [IV] which was used in the next step directly. MS (ESI): m / z = 358.0 [M+H]+.

[0119] Step 3:

[0120] To a solution of Intermediate [IV] (1000 mg, 1 Eq, 2.798 mmol) and O-(tetrahydro-2H-pyran-2- yl)hydroxylamine (393.4 mg, 1.2 Eq, 3.358 mmol) in DMF (15 mL) was added HATU (1.595 g, 1.5 Eq, 4.197 mmol), and DIPEA (1.805 g, 2.43 mL, 5 Eq, 13.99 mmol). The mixture was stirred at 25 °C for 1 hour. Then, the reaction was washed with NaHCOa aq., extracted with CHCla / iPrOH (4: 1), dried over anhydrous NaaSCV, concentrated, and purified with CombiFlash chromatography using MeOH and 0 to 5% dichloromethane as eluent to provide Intermediate {VI] (787 mg, 61.6%). MS (ESI): m / z = 479.1 [M+Na]+.

[0122] To a solution of Intermediate [VI] (500 mg, 1 Eq, 1 .10 mmol) in MeOH (2 mL) and water (2 mL) was added TFA (0.2 mL), and the mixture was stirred at 25 °C for 2 hour. The reaction mixture was then purified by Pre-HPLC to provide compound 41 (197 mg, 48.3%).1H NMR (500 MHz, DMSO-d6) 5 10.35 (d, J = 1.7 Hz, 1 H), 8.72 (d, J = 1 .7 Hz, 1 H), 8.52 (dd, J = 7.2, 1 .1 Hz, 2H), 8.48 (dd, J = 8.3, 1 .2 Hz, 2H), 7.89 (dd, J = 8.2, 7.3 Hz, 2H), 4.25 (t, J = 6.5 Hz, 2H), 3.66 (t, J = 6.5 Hz, 2H), 3.58 - 3.52 (m, 4H), 3.45 (dd, J = 5.8, 3.8 Hz, 2H), 2.12 (t, J = 6.3 Hz, 2H). MS (ESI): m / z = 372.9 [M+H]+.

[0123] The title compound was prepared according to the same procedure as Comparator Compound 13. Purification by preparative reverse-phase HPLC (0- 95% MeOH in water, 25min, 20 mL / min) afforded the product as a white powder (12 mg, 4 steps yield 46%).1H NMR (500 MHz, DMSO-d6) 5 10.35 (s, 1 H), 8.72 (s, 1 H), 8.51 (dd, J = 7.2, 1 .2 Hz, 2H), 8.47 (dd, J = 8.3, 1 .1 Hz, 2H), 7.88 (dd, J = 8.2, 7.3 Hz, 2H), 4.22 (t, J = 6.7 Hz, 2H), 3.65 (t, J = 6.4 Hz, 2H), 3.61 (t, J = 6.7 Hz, 2H), 2.17 (t, J = 6.4 Hz, 2H). MS (ESI): m / z = 328.90. [M+H]+.

[0124] The title compound was prepared according to the same procedure with Comparator Compound 16. Purification by preparative reverse-phase HPLC (0- 95% MeOH in water, 25min, 20 mL / min) afforded the product as a white powder (13 mg, 4 steps yield 40%).1H NMR (500 MHz, DMSO-d6) 5 10.35 (s, 1 H), 8.51 (dd, J = 7.2, 1 .1 Hz, 2H), 8.47 (dd, J = 8.4, 1 .2 Hz, 2H), 7.88 (dd, J = 8.2, 7.2 Hz, 2H), 4.25 (t, J = 6.4 Hz, 2H), 3.66 (t, J = 6.4 Hz, 2H), 3.57 - 3.53 (m, 2H), 3.51 (t, J = 6.3 Hz, 2H), 3.48 - 3.44 (m, 2H), 3.41 (dd, J = 5.9, 3.7 Hz, 2H), 3.34 (dd, J = 5.9, 3.7 Hz, 2H), 2.15 (d, J = 6.3 Hz, 2H). MS (ESI): m / z = 416.95 [M+H]+.Compound 43

[0125] To a solution of 1 H,3H-benzo[de]isochromene-1 , 3-dione (534 mg, 1.2 Eq, 2.69 mmol) in DMF (2 mL) was added 4-aminobutan-1-ol (200 mg, 1 Eq, 2.24 mmol) and the mixture was stirred at 125 °C for 12 hour. Removed the solvent and purified with Combi Flash (MeOH / DCM 0 to 10%) affording the desired product (506 mg, 83.7%). MS (ESI): m / z = 284.06 [M+H]+.

[0126] To a solution of 2-(5-hydroxypentyl)-1 H-benzo[de]isoquinoline-1 ,3(2H)-dione (100 mg, 1 Eq, 353 pmol) K2CO3 (244 mg, 5 Eq, 1.76 mmol) in DMF (2 mL) was added ferf-butyl acrylate (136 mg, 155 pL, 3 Eq, 1.06 mmol) and the mixture was stirred at 90 °C for 12 hour. Filtered and then removed the solvent and purified by preparative reverse-phase HPLC (45- 95% MeOH in water, 56min, 40 mL / min) afforded the product as white powder (17 mg, yield 12%). MS (ESI): m / z = 434.18 [M+Na]+.

[0127] To a solution of ferf-butyl 3-((5-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)pentyl)oxy)propanoate (15 mg, 1 Eq, 36 pmol) in DCM (2 mL) was added TFA (1 mL, 1 Eq, 36 pmol) and the mixture was stirred at 25 °C for 1 hour. Then remove the solvents and put them into the next step directly. MS (ESI): m / z = 356.07 [M+H]+.

[0128] To a solution of 3-((5-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)pentyl)oxy)propanoic acid (13 mg, 1 Eq, 37 pmol) in DMF (2 mL) was added HATU (21 mg, 1.5 Eq, 55 pmol) and DIPEA (24 mg, 32 pL, 5 Eq, 0.18 mmol)and the mixture was stirred at 25 °C for 30 min. Then hydroxylamine hydrochloride (5.1 mg, 2 Eq, 73 pmol) was added and stirred for another 30 min. Purification by preparative reverse-phase HPLC (10- 95%MeOH in water, 25min, 20 mL / min) afforded the product as a white powder (4 mg, 2 steps yield 30%).1H NMR (500 MHz, DMSO-de) 5 10.39 (s, 1 H), 8.50 (ddd, J = 19.5, 7.8, 1.1 Hz, 4H), 7.89 (dd, J = 8.2, 7.2 Hz, 2H), 4.08 - 4.01 (m, 2H), 3.55 (t, J = 6.3 Hz, 2H), 3.35 (t, J = 6.6 Hz, 2H), 2.17 (t, J = 6.3 Hz, 2H), 1 .69 - 1 .58 (m, 2H), 1 .53 (q, J = 7.1 Hz, 2H), 1.42 - 1.31 (m, 2H). MS (ESI): m / z = 371.12 [M+H]+.Compound 44

[0129] To a solution of 2-aminoethan-1-ol (200 mg, 1 Eq, 3.27 mmol) in DMF (2 mL) was added 1 H,3H- benzo[de]isochromene-1, 3-dione (779 mg, 1.2 Eq, 3.93 mmol) and the mixture was stirred at 125 °C for 12 hour. Removed the solvent and purified with CombiFlash (MeOH / DCM 0 to 10%) affording the desired product (216.3 mg, 27.4%). MS (ESI): m / z = 241.97 [M+H]+.

[0130] The aqua solution of NaOH (381 mg, 23 Eq, 9.53 mmol, dissolved in 2 mL) was added to a solution of 2-(2-hydroxyethyl)-1 H-benzo[de]isoquinoline-1 ,3(2H)-dione (100 mg, 1 Eq, 415 pmol) and TBAB (13.4 mg, 0.1 Eq, 41.5 pmol) in toluene (6 ml). The mixture was stirred at r.t. for 10 min., then ferf-butyl 6-bromohexanoate (208 mg, 2 Eq, 829 pmol) was added, and the reaction was stirred at 60 °C. for 12 h. The mixture was diluted with EtOAc (100 ml), and washed with water (3x50 ml), the organic layers were dried over anhydrous Na2SO4, and the solvents were removed under reduced pressure. The residue was purified by preparative reverse-phase HPLC (45- 95% MeOH in water, 56min, 40 mL / min) afforded the product as a white powder (40.0 mg, 23.0%). MS (ESI): m / z = 434.18 [M+Na]+.

[0131] To a solution of ferf-butyl 6-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)hexanoate (20 mg, 1 Eq, 49 pmol) in DCM (3 mL) was added TFA (1 mL, 1 Eq, 49 pmol) and the mixture was stirred at 25 °C for 30 min. Then remove the solvents and put them into the next step directly. MS (ESI): m / z = 356.02 [M+H]+.

[0132] To a solution of 6-(2-(1,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)hexanoic acid (17 mg, 1 Eq, 48 pmol) and HATU (27 mg, 1.5 Eq, 72 pmol) in DMF (2 mL) was added DIPEA (31 mg, 42 pL, 5 Eq, 0.24 mmol) and the mixture was stirred at 25 °C for 5 min. Then hydroxylamine hydrochloride (6.6 mg, 2 Eq, 96 pmol) was added dropwise and stirred for 30 min. Purification by preparative reverse-phase HPLC (10- 95% MeOH in water, 25min, 20 mL / min) afforded the product as a white powder (5 mg, 2 steps yield 30%).1H NMR (500 MHz, DMSO-cfe) 5 10.30 (s, 1 H), 8.49 (ddd, J = 19.5, 7.8, 1.1 Hz, 4H), 7.88 (dd, J = 8.2, 7.3 Hz, 2H), 4.24 (t, J = 6.5 Hz, 2H), 3.61 (t, J = 6.5 Hz, 2H), 3.40 (t, J = 6.6 Hz, 2H), 1 .85 (t, J = 7.4 Hz, 2H), 1 .47 - 1.37 (m, 4H), 1.29 - 1.14 (m, 2H). MS (ESI): m / z = 371.27 [M+H]+.Compound 45

[0133] To a solution of tert-butyl (2-(2-aminoethoxy)ethyl)(methyl)carbamate (70 mg, 1 Eq, 0.32 mmol) in DMF (2 mL) was added 1 H,3H-benzo[de]isochromene-1 , 3-dione (0.13 g, 2 Eq, 0.64 mmol) and the mixture was stirred at 125 °C for 12 hour. Removed the solvent and purified with CombiFlash (MeOH / DCM 0 to 5%) to afford the product (91.8mg, 72%).

[0134] To a solution of tert-butyl (2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethyl)(methyl)carbamate (91 mg, 1 Eq, 0.23 mmol) in DCM (2 mL) was added TFA (0.5 mL, 1 Eq, 0.23 mmol) and the mixture was stirred at 25 °C for 30 min. Removed the solvent and put it into the next step directly.

[0135] To a solution of 2-(2-(2-(methylamino)ethoxy)ethyl)-1 H-benzo[de]isoquinoline-1 ,3(2H)-dione (68 mg, 1 Eq, 0.23 mmol) and K2CO3 (94 mg, 3 Eq, 0.68 mmol) in DMF (2 mL) was added tert-butyl 3-bromopropanoate (71 mg, 57 pL, 1.5 Eq, 0.34 mmol) and the mixture was stirred at 100 °C for 24 hour. Filtered, removed the solvent, and purified with CombiFlash (MeOH / DCM 0 to 10%) to afford the product (80.4, 83%).

[0136] To a solution of tert-butyl 3-((2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethyl)(methyl)amino)propanoate (40 mg, 1 Eq, 94 pmol) in DCM (2 mL) was added TFA (1 mL, 1 Eq,94 pmol) and the mixture was stirred at 25 °C for 1 hour. Removed the solvent and put it into the next step directly.

[0137] To a solution of 3-((2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethyl)(methyl)amino)propanoic acid (35 mg, 1 Eq, 94 pmol) and HATU (72 mg, 2 Eq, 0.19 mmol) in DMF (2 mL) was added DIPEA (61 mg, 5 Eq, 0.47 mmol) and O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (17 mg, 1.5 Eq, 0.14 mmol) and the mixture was stirred at 25 °C for 30 min. Extracted with CHClaZ / PrOH (4:1), dried over anhydrous Na2SO4, removed solvents, and then purified with CombiFlash (MeOH / DCM 0 to 10%) affording product (15 mg, 34%).

[0138] To a solution of 3-((2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethyl)(methyl)amino)-N- ((tetrahydro-2H-pyran-2-yl)oxy)propanamide (15 mg, 1 Eq, 32 pmol) in MeOH (2 mL) and H2O (1 mL) was added TFA (0.5 mL, 1 Eq, 32 pmol) and the mixture was stirred at 25 °C for 1 hour. Purified by pre-HPLC, affording the product (6.3mg, 51 %).1H NMR (500 MHz, DMSO-d6) 5 10.64 (s, 1 H), 8.51 (ddd, J = 14.4, 7.8, 1.1 Hz, 4H), 7.90 (dd, J = 8.2, 7.2 Hz, 2H), 4.31 (td, J = 6.1 , 1.7 Hz, 2H), 3.84 - 3.78 (m, 2H), 3.76 (t, J = 6.0 Hz, 2H), 3.37 (td, J = 15.4, 7.5 Hz, 2H), 3.23 (dt, J = 13.1 , 6.1 Hz, 2H), 2.74 (d, J = 4.2 Hz, 3H), 2.42 (td, J = 7.3, 1.9 Hz, 2H). MS (ESI): m / z = 386.07 [M+H]+.Compound 46

[0139] Compound 46 was synthesized with the same procedure as Compound 45.

[0140] 1H NMR (500 MHz, DMSO-d6) 5 10.45 (s, 1 H), 9.30 (s, 1 H), 8.53 (ddd, J = 12.5, 7.8, 1.1 Hz, 4H), 7.91 (dd, J = 8.2, 7.3 Hz, 2H), 4.47 - 4.37 (m, 2H), 3.73 (q, J = 4.2 Hz, 2H), 3.66 (t, J = 6.3 Hz, 2H), 3.55 (tt, J = 14.2, 4.7 Hz, 2H), 3.41 (ddd, J = 33.0, 13.5, 6.1 Hz, 2H), 2.92 (d, J = 4.4 Hz, 3H), 2.23 (td, J = 6.3, 2.2 Hz, 2H).MS (ESI): m / z = 386.07 [M+H]+.Compound 47

[0141] Compound 47 was synthesized with the same procedure as Compound 45.

[0142] 1H NMR (500 MHz, DMSO-d6) 5 10.37 (s, 1 H), 9.15 (s, 1 H), 8.53 (ddd, J = 12.6, 7.8, 1.2 Hz, 4H), 7.91 (dd, J = 8.2, 7.2 Hz, 2H), 4.48 - 4.34 (m, 2H), 3.55 - 3.47 (m, 2H), 3.40 (dq, J = 12.4, 6.0 Hz, 2H), 3.34 - 3.04 (m, 2H), 2.97 - 2.86 (m, 3H), 1 .96 (t, J = 7.3 Hz, 2H), 1 .54 (tt, J = 15.2, 7.2 Hz, 2H), 1 .33 - 1 .22 (m, 2H). MS (ESI): m / z = 384.12 [M+H]+.Compound 48

[0143] Compound 48 was synthesized with the same procedure as Compound 44.

[0144] 1H NMR (500 MHz, DMSO-d6) 5 10.37 (s, 1 H), 8.51 (dd, J = 7.3, 1.2 Hz, 2H), 8.48 (dd, J= 8.3, 1.1 Hz, 2H), 7.88 (dd, J = 8.3, 7.2 Hz, 2H), 3.65 (t, J = 6.3 Hz, 2H), 3.47 - 3.39 (m, 4H), 2.17 (t, J = 6.3 Hz, 2H). MS (ESI): m / z = 398.9 [M+Na]+.Compound 49

[0145] To a solution of 5-nitro-1 H,3H-benzo[de]isochromene-1, 3-dione (115 mg, 1.1 Eq, 471 pmol)tert-butyl 3- (2-(2-aminoethoxy)ethoxy)propanoate (100 mg, 1 Eq, 429 pmol) in DMF (5 mL) was stirred at 125 °C for 5 hours. Removed the solvents and purified with CombiFlash (MeOH / DCM 0-10%), giving the product (180mg, 91.6%).

[0146] To a solution of tert-butyl 3-(2-(2-(5-nitro-1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethoxy)propanoate (180 mg, 1 Eq, 393 pmol) in DCM (5 mL) was added TFA (1 mL) and the mixture was stirred at 25 °C for 1 hour. Then removed the solvents and put them into the next step directly. To a solution of the above acid and HATU (194 mg, 1.3 Eq, 510 pmol) in DMF (5 mL) was added DIPEA (253 mg, 341 pL, 5 Eq, 1.96 mmol) and O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (55.2 mg, 1.2 Eq, 471 pmol) and then the mixture was stirred at 25 °C for 1 hour. Purified with CombiFlash (MeOH / DCM 0-10%) affording the product (121 mg, 61.5%).

[0147] To a solution of the 3-(2-(2-(5-nitro-1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)-N- ((tetrahydro-2H-pyran-2-yl)oxy)propanamide (25 mg, 1 Eq, 50 pmol) in MeOH (4 mL) and H2O (1 mL) was added TFA (0.5 mL), then stirred at 25 °C for 2h. Removed solvents and purified with Pre-HPLC, affording the product (13mg, 62%).1H NMR (500 MHz, DMSO-d6) 5 10.33 (s, 1 H), 9.49 (d, J = 2.3 Hz, 1 H), 8.97 (d, J = 2.2 Hz, 1 H), 8.79 (dd, J = 8.4, 1.2 Hz, 1 H), 8.69 (dd, J = 7.3, 1.1 Hz, 1 H), 8.10 - 8.03 (m, 1 H), 4.26 (t, J = 6.4 Hz, 2H), 3.67 (t,J = 6.4 Hz, 2H), 3.62 - 3.47 (m, 4H), 3.44 (dd, J = 5.9, 3.8 Hz, 2H), 2.10 (t, J = 6.3 Hz, 2H). MS (ESI): m / z = 417.95 [M+H]+.

[0148] To a solution of ferf-butyl 3-(2-(2-aminoethoxy)ethoxy)propanoate (500 mg, 1 Eq, 2.14 mmol) in DMF (5 mL) was added 6-nitro-1 H,3H-benzo[de]isochromene-1, 3-dione (573 mg, 1.1 Eq, 2.36 mmol), then stirred at 125 °C for 5 hour. Removed the solvents and purified with CombiFlash (MeOH / DCM 0-10%), giving the product (874mg, 89.0%).

[0149] To a solution of ferf-butyl 3-(2-(2-(6-nitro-1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethoxy)propanoate (100 mg, 1 Eq, 218 pmol) in DCM (5 mL) was added TFA (0.5 mL, 1 Eq, 218 pmol) and the mixture was stirred at 25 °C for 1 hour. Removed the solvent and put it into the next step directly.

[0150] To a solution of 3-(2-(2-(6-nitro-1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)propanoic acid (87 mg, 1 Eq, 0.22 mmol) and HATU (0.16 g, 2 Eq, 0.43 mmol) in DMF (2 mL) was added DIPEA (0.14 g, 0.19 mL, 5 Eq, 1.1 mmol) and hydroxylamine hydrochloride (20 mg, 1.3 Eq, 0.28 mmol), then the mixture was stirred at 25 °C for 30 min. Purified by pre-HPLC affording the product (80mg, 89%) as a white powder.1H NMR (500 MHz, DMSO-d6) 5 10.36 (s, 1 H), 8.73 (dd, J = 8.7, 1.1 Hz, 1 H), 8.65 (dd, J = 7.3, 1.1 Hz, 1 H), 8.63 (d, J = 8.0 Hz, 1 H), 8.57 (d, J = 8.0 Hz, 1 H), 8.11 (dd, J = 8.7, 7.3 Hz, 1 H), 4.25 (t, J = 6.5 Hz, 2H), 3.67 (t, J = 6.5 Hz, 2H), 3.58 - 3.50 (m, 4H), 3.47 - 3.36 (m, 2H), 2.12 (t, J = 6.3 Hz, 2H). MS (ESI): m / z = 418.12 [M+H]+.51

[0151] To a solution of 3-(2-(2-(5-nitro-1,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)-N- ((tetrahydro-2H-pyran-2-yl)oxy)propanamide (80 mg, 1 Eq, 0.16 mmol) in MeOH (5 mL) was added Pd / C (15 mg) and purged with H2 three times and the mixture was stirred at 25 °C for 3 hour. Filtered, removed the solvent, and put it into the next step directly.

[0152] To a solution of the 3-(2-(2-(5-amino-1,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)-N- ((tetrahydro-2H-pyran-2-yl)oxy)propanamide (20 mg, 1 Eq, 42 pmol) in MeOH (4 mL) and H2O (1 mL) was added TFA (0.5 mL), then stirred at 25 °C for 2h. Removed solvents and purified with Pre-HPLC, affording the product (10mg, 61 %).1H NMR (500 MHz, DMSO-de) 3 10.35 (s, 1 H), 8.09 (dd, J = 7.2, 1.1 Hz, 1 H), 8.05 (dd, J = 8.4, 1.1 Hz, 1 H), 7.98 (d, J = 2.3 Hz, 1 H), 7.62 (dd, J = 8.3, 7.2 Hz, 1 H), 7.31 (d, J = 2.3 Hz, 1 H), 4.21 (t, J = 6.5 Hz, 2H), 3.62 (t, J = 6.5 Hz, 2H), 3.58 - 3.49 (m, 4H), 3.44 (dd, J = 5.8, 3.8 Hz, 2H), 2.12 (t, J = 6.3 Hz, 2H). MS (ESI): m / z = 387.95[M+H]+.

[0153] Compound 52 was synthesized with the same procedure as Compound 51 .

[0154] 1H NMR (500 MHz, DMSO-de) 3 10.30 (s, 1 H), 8.55 (dd, J = 8.4, 1.2 Hz, 1 H), 8.42 - 8.32 (m, 1 H), 8.12 (d, J = 8.4 Hz, 1 H), 7.64 - 7.55 (m, 1 H), 7.40 (s, 2H), 6.77 (d, J = 8.4 Hz, 1 H), 4.12 (t, J = 6.6 Hz, 2H), 3.53 (t, J = 6.6 Hz, 2H), 3.50 - 3.44 (m, 4H), 3.39 - 3.36 (m, 2H), 2.06 (t, J = 6.3 Hz, 2H). MS (ESI): m / z = 388.12 [M+H]+.Compound 53

[0155] To a solution of compound 1 (10.0 g, 62.1 mmol, 1.0 eq) in DCM (50 mL) were added compound 2 (9.25 g, 62.1 mmol, 1.0 eq), NaOH (2.48 g, 62.0 mmol, 1.0 eq) and TBAI (22.9 g, 62.1 mmol, 1.0 eq) in water (10 mL). The reaction mixture was stirred at 50 °C for 16 hours. LCMS and TLC showed the formation of compound 3. The mixture was filtered and the filtrate was concentrated. The crude product was purified by flash column chromatography (PE / EA, 3 / 1) to afford compound 3 (5.0 g, 35%) as colorless oil. LCMS: 230.00 [M+H]+

[0156] To a solution of compound 3 (5.00 g, 21.8 mmol, 1.0 eq) in THF (50 mL) were added NalO4 (4.69 g, 21.7 mmol, 1.0 eq) and KOSO42H2O (6.68 g, 21.5 mmol, 1.0 eq) and water (10 mL). The reaction mixture was stirred at 50 °C for 16 hours. LCMS and TLC showed the formation of compound 4. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic layer was washed with water and brine, and concentrated. The crude product was purified by flash column chromatography (PE / EA, 3 / 1) to afford compound 4 (4.20 g, 83%) as colorless oil. LCMS: 232.00 [M+H]+

[0157] To a solution of compound 4 (4.20 g, 18.2 mmol, 1.0 eq) in THF (20 mL) was added compound 5 (8.10 g, 21 .5 mmol, 1 .2 eq). The mixture was heated at 60 °C for 2 hours. The mixture was concentrated and the crude product was purified by flash column chromatography (PE / EA, 10 / 1) to afford compound 6 (4.2 g, 70%) as yellow oil. LCMS: 330.30 [M+H]+

[0158] To a solution of compound 6 (4.20 g, 12.8 mmol, 1.0 eq) in dioxane (4 mL) was added HCI solution (4 M in dioxane, 25 mL). Then the solution was stirred at rt for 1 hour. The reaction solution was concentrated to afford compound 7 (2.8 g, crude) as yellow oil, which was used in the next step without further purification. LCMS: 174.15 [M+H]+

[0159] To a solution of compound 7 (2.80 g, crude) in DMSO (20 mL) were added DIEA (1 .60 g, 12.4 mmol), compound 8 (2.40 g, 12.1 mmol). The reaction mixture was heated at 70 °C for 1 hour. The reaction solution was partitioned between water (50 mL) and ethyl acetate (100 mL x 3). The organic layer was concentrated and purified by flash column chromatography (PE / EA, 1 / 1 ) to afford compound 9 (1.20 g, 27% over two steps) as yellow oil. LC-MS: 354.20 [M+H]+

[0160] To a solution of compound 9 (600 mg, 1.70 mmol, 1.0 eq) in ACN (10 mL) were added TCFH (523 mg, 1.87 mmol, 1.1 eq), NMI (418 mg, 5.10 mmol, 3.0 eq) and compound 10 (298 mg, 2.55 mmol, 1.5 eq). The mixture was stirred at room temperature for 16 h. The reaction was completed (detected by TLC). The reaction was quenched with water (10 mL) and extracted with ethyl acetate (50 mL). The organic layer was concentrated and purified by flash column chromatography (PE / EA, 3 / 1) to afford compound 11 (230 mg, 30%) as white solid. LC-MS: 453.50 [M+H]+

[0161] A solution of the compound 11 (230 mg, 0.508 mmol, 1.0 eq) in ethanol (3 mL) was added TsOH'hhO (17 mg, 0.10 mmol, 0.2 eq). The reaction mixture was allowed to stir at room temperature for 16 hours. The precipitate from the reaction was purified by prep-HPLC (5% to 95% ACN in water) to afford 53 (103 mg, 53%) as light-yellow solid. LC-MS: 387.00 [M+H]+ 1H NMR (400 MHz, DMSO-d6) 5 10.48 (brs, 1 H), 8.48 (d, J = 7.7 Hz, 2H), 8.44 (d, J = 7.7 Hz, 2H), 7.85 (t, J = 7.7 Hz, 2H), 6.62 - 6.49 (m, 1 H), 5.67 (d, J = 15.5 Hz, 1 H), 4.22 (t, J = 6.4 Hz, 2H), 3.60 (t, J = 6.4 Hz, 2H), 3.43 - 3.37 (m, 2H), 2.12 - 2.04 (m, 2H), 1 .57 - 1 .47 (m, 2H).Comparator Compound 1

[0162] To a solution of 3-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)propanoic acid (150 mg, 1 Eq, 420 pmol) in DMF (2 mL) was added HATU (191 mg, 1.2 Eq, 504 pmol) DIPEA (162 mg, 219 pL, 3 Eq, 1.26 mmol)and the mixture was stirred at 25 °C for 30 min. Then NH3 (89.9 pL, 7 M in MeOH, 1.5 Eq, 630 pmol) was added dropwise and the mixture was stirred at RT for 30 minutes. Purification by preparative reverse-phase HPLC (1- 80% MeOH in water, 56min, 40 mL / min) afforded the product as a white powder (64 mg, 43%).1H NMR (500 MHz, DMSO-d6) 5 8.49 (ddd, J = 21 .8, 7.8, 1 .1 Hz, 4H), 7.88 (dd, J = 8.3, 7.2 Hz, 2H), 7.25 (s, 1 H), 6.75 (s, 1 H), 4.25 (t, J = 6.5 Hz, 2H), 3.66 (t, J = 6.5 Hz, 2H), 3.59 - 3.49 (m, 4H), 3.47 - 3.42 (m, 2H), 2.20 (t, J = 6.5 Hz, 2H). MS (ESI): m / z = 356.95 [M+H]+.Comparator Compound 2Comp 2

[0163] To a solution of 3-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)propanoic acid (40 mg, 1 Eq, 0.11 mmol), tert-butyl (2-aminophenyl)carbamate (55 mg, 64% Wt, 1.5 Eq, 0.17 mmol), HATU (85 mg, 2 Eq, 0.22 mmol) in DMF (2 mL) was added DIPEA (72 mg, 97 pL, 5 Eq, 0.56 mmol) and the mixture was stirred at 25 °C for 30 min. Extracted with CHClaZ / PrOH (4:1), dried over anhydrous Na2SO4, removed solvents, and then purified with CombiFlash (MeOH / DCM 0 to 5%) affording product (55 mg, 90%).

[0164] To a solution of tert-butyl (2-(3-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethoxy)propanamido)phenyl)carbamate (30 mg, 1 Eq, 55 pmol) in DCM (2 mL) was added TFA (1 mL, 1 Eq, 55 pmol) and the mixture was stirred at 25 °C for 30 min. Then the solvent was removed and the resulting residue was dissolved in DMSO (2mL). Purified by Pre-HPLC, affording the product as a white powder (18.1 mg, 74%).1H NMR (500 MHz, DMSO-d6) 5 9.51 (s, 1 H), 8.55 - 8.45 (m, 4H), 7.92 - 7.85 (m, 2H), 7.19 (dd, J = 7.9, 1 .5 Hz, 1 H), 7.08 - 7.01 (m, 1 H), 6.95 (d, J = 7.9 Hz, 1 H), 6.84 (d, J = 7.9 Hz, 1 H), 4.26 (t, J = 6.5 Hz, 2H), 3.67 (td, J = 6.5, 3.2 Hz, 4H), 3.61 - 3.55 (m, 2H), 3.52 (dd, J = 5.9, 3.6 Hz, 2H), 2.54 (d, J = 6.4 Hz, 2H). MS (ESI): m / z = 448.13 [M+H]+.Comparator Compound 3

[0165] To a solution of 3-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)propanoic acid (15 mg, 1 Eq, 42 pmol), HATU (24 mg, 1.5 Eq, 63 pmol) in DMF (2 mL) was added DIPEA (27 mg, 36 pL, 5 Eq, 0.21 mmol) and N-Methylhydroxylamine hydrochloride (6.6 mg, 1.2 Eq, 50 pmol) the mixture was stirred at 25 °C for 30 min. Purification by pre-HPLC affords the product (8.7mg, 54%).1H NMR (500 MHz, DMSO-de) 5 9.80 (s, 1 H), 8.52 (dd, J = 7.2, 1 .2 Hz, 2H), 8.48 (dd, J = 8.3, 1 .2 Hz, 2H), 7.89 (dd, J = 8.3, 7.3 Hz, 2H), 4.25 (t, J = 6.5 Hz,2H), 3.66 (t, J = 6.5 Hz, 2H), 3.58 - 3.51 (m, 4H), 3.50 - 3.43 (m, 2H), 3.05 (s, 3H). MS (ESI): m / z = 387.07 [M+H]+.Comp 4

[0166] To a solution of 3-(2-(2-(1,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)propanoic acid (30 mg, 1 Eq, 84 pmol), tert-butyl hydrazinecarboxylate (13 mg, 1.2 Eq, 0.10 mmol), HATU (64 mg, 2 Eq, 0.17 mmol)in DMF (2 mL) was added DIPEA (54 mg, 73 pL, 5 Eq, 0.42 mmol) and the mixture was stirred at 25 °C for 30 min. Extracted with CHClaZ / PrOH (4: 1), dried over anhydrous Na2SC>4, removed solvents, and then purified with CombiFlash (MeOH / DCM 0 to 5%) affording the product (36mg, 91 %).

[0167] To a solution of tert-butyl 2-(3-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethoxy)propanoyl)hydrazine-1 -carboxylate (30 mg, 1 Eq, 64 pmol) in DCM (2 mL) was added TFA (1 mL, 1 Eq, 64 pmol) and the mixture was stirred at 25 °C for 30 min. Then the solvent was removed, and the resulting residue was dissolved in DMSO (2mL) and purified by pre-HPLC, affording the product as a white powder (13.6mg, 58%).1H NMR (500 MHz, DMSO-d6) 3 10.68 (s, 1 H), 8.51 (dd, J = 7.3, 1.2 Hz, 2H), 8.48 (dd, J = 8.3, 1.1 Hz, 2H), 7.89 (dd, J = 8.2, 7.2 Hz, 2H), 4.25 (t, J = 6.5 Hz, 2H), 3.65 (t, J = 6.5 Hz, 2H), 3.60 (t, J = 6.0 Hz, 2H), 3.54 (dd, J = 5.9, 3.6 Hz, 2H), 3.47 (dd, J = 5.9, 3.6 Hz, 2H), 2.38 (t, J = 6.1 Hz, 2H). MS (ESI): m / z = 372.07 [M+H]+.Comparator Compound 5

[0168] To a solution of 3-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)propanehydrazide (56 mg, 1 Eq, 0.15 mmol) in DCM (2 mL) was added 2,2-difluoroacetic anhydride (26 mg, 19 pL, 1 Eq, 0.15 mmol) at 0 °C and the mixture was stirred at 0 °C for 2 hour. Then NaHCOa aq. solution work up, extracted with CHCla / IPrOH (4: 1), dried over anhydrous Na2SO4, remove solvents and then purify with pre-HPLC affording product (14mg, 21 %).1H NMR (500 MHz, DMSO-d6) 3 10.86 (d, J = 1.8 Hz, 1 H), 10.12 (d, J = 1.7 Hz, 1 H), 8.51(dd, J = 7.3, 1 .2 Hz, 2H), 8.47 (dd, J = 8.3, 1 .1 Hz, 2H), 7.88 (dd, J = 8.3, 7.3 Hz, 2H), 6.33 (t, J = 53.0 Hz, 1 H), 4.25 (t, J = 6.5 Hz, 2H), 3.65 (t, J = 6.5 Hz, 2H), 3.58 (t, J = 6.4 Hz, 2H), 3.54 (dd, J = 5.9, 3.7 Hz, 2H), 3.46 (dd, J = 5.9, 3.8 Hz, 2H), 2.34 (t, J = 6.4 Hz, 2H). MS (ESI): m / z = 450.08 [M+H]+.Comparator Compound 6Comp. 6

[0169] To a solution of 1 H,3H-benzo[de]isochromene-1 , 3-dione (500 mg, 1.0 Eq, 2.52 mmol) in DMF (5 mL) was tert-butyl (2-(2-(2-aminoethoxy)ethoxy)ethyl)carbamate (752 mg, 1.2 Eq, 3.03 mmol), and stirred at 125 °C for 24 hour. Removed the solvent and purified with CombiFlash (MeOH / DCM 0 to 5%) to afford the product (897mg, 83.0%).

[0170] To a solution of tert-butyl (2-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethoxy)ethyl)carbamate (80 mg, 1 Eq, 0.19 mmol) in DCM (2 mL) was added TFA (1 mL, 1 Eq, 0.19 mmol) and the mixture was stirred at 25 °C for 30 min. Removed the solvent under vacuum and put it into the next step directly.

[0171] To a solution of the above residue, 2-(methylamino)-2-oxoacetic acid (23 mg, 1.2 Eq, 0.22 mmol), HATU (0.11 g, 1.5 Eq, 0.28 mmol) in DMF (1 mL) was added DIPEA (96 mg, 0.13 mL, 4 Eq, 0.75 mmol) and the mixture was stirred at 25 °C for 30 min. Purified by pre-HPLC to afford the desired product (59.4mg, 77%).1H NMR (500 MHz, DMSO-d6) 5 8.69 (q, J = 5.0 Hz, 1 H), 8.56 (t, J = 6.0 Hz, 1 H), 8.52 (dd, J = 7.2, 1 .2 Hz, 2H), 8.48 (dd, J = 8.3, 1.1 Hz, 2H), 7.88 (dd, J = 8.2, 7.2 Hz, 2H), 4.26 (t, J = 6.4 Hz, 2H), 3.67 (t, J = 6.5 Hz, 2H), 3.56 (dd, J = 5.9, 3.5 Hz, 2H), 3.49 (dd, J = 5.9, 3.6 Hz, 2H), 3.42 (t, J = 6.0 Hz, 2H), 3.22 (q, J = 6.0 Hz, 2H), 2.63 (d, J = 4.9 Hz, 3H). MS (ESI): m / z = 414.12 [M+H]+.Comparator Compound 7

[0172] To a solution of tert-butyl (2-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethoxy)ethyl)carbamate (70 mg, 1 Eq, 0.16 mmol) in DCM (2 mL) was added TFA (1 mL, 1 Eq, 0.16 mmol) and the mixture was stirred at 25 °C for 30 min. Removed the solvent and put it into the next step directly.

[0173] To a solution of the above residue, DIPEA (84 mg, 0.11 mL, 4 Eq, 0.65 mmol) in DMF (2 mL) was added sulfamoyl chloride (23 mg, 1.2 Eq, 0.20 mmol), and the mixture was stirred at 0 °C for 60 min. Purified by pre-HPLC to afford the desired product (6mg, 9%).1H NMR (500 MHz, DMSO-d6) 5 8.53 (dd, J = 7.2, 1.1 Hz, 2H), 8.49 (dd, J = 8.2, 1.1 Hz, 2H), 7.89 (t, J = 7.7 Hz, 2H), 6.51 (s, 2H), 6.43 (t, J = 6.1 Hz, 1 H), 4.26 (t, J = 6.5 Hz, 2H), 3.67 (t, J = 6.5 Hz, 2H), 3.58 (dd, J = 5.9, 3.7 Hz, 2H), 3.49 (dd, J = 5.9, 3.7 Hz, 2H), 3.44 (t, J = 6.2 Hz, 2H), 2.98 (q, J = 6.2 Hz, 2H). MS (ESI): m / z = 430.08 [M+Na]+.Comparator Compound 8

[0174] To a solution of tert-butyl (2-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethoxy)ethyl)carbamate (100 mg, 1 Eq, 233 pmol) in DCM (2 mL) was added TFA (1 mL, 1 Eq, 233 pmol) and the mixture was stirred at 25 °C for 30 min. Then remove the solvent and put it into the next step directly.

[0175] To a solution of 2-(acetylthio) acetic acid (37.6 mg, 29 pL, 1.2 Eq, 280 pmol) and above amine in DCM (2 mL) was added HATU (133 mg, 1.5 Eq, 350 pmol) and DIPEA (120 mg, 162 pL, 4 Eq, 934 pmol), and the mixture was stirred at 25 °C for 30 min. NaHCOa aq. solution workup, extracted with CHCla / IPrOH (4: 1), dried over anhydrous Na2SO4, removed solvents, and then purified by preparative reverse-phase HPLC afforded the product.1H NMR (500 MHz, DMSO-d6) 5 8.51 (dd, J = 7.2, 1.1 Hz, 2H), 8.47 (dd, J = 8.4, 1.1 Hz, 2H), 8.12 (t, J = 5.6 Hz, 1 H), 7.88 (dd, J = 8.2, 7.3 Hz, 2H), 4.25 (t, J = 6.5 Hz, 2H), 3.66 (t, J = 6.5 Hz, 2H), 3.59 - 3.53 (m, 4H), 3.50 - 3.44 (m, 2H), 3.34 (t, J = 5.8 Hz, 2H), 3.11 (q, J = 5.8 Hz, 2H), 2.32 (s, 3H). MS (ESI): m / z = 445.08 [M+H]+.Comparator Compound 9

[0176] To a solution of S-(2-((2-(2-(2-(1,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl)ethoxy)ethoxy)ethyl)amino)- 2-oxoethyl) ethanethioate (60 mg, 1 Eq, 0.13 mmol) in MeOH (2 mL) was added potassium carbonate (22 mg, 1.2 Eq, 0.16 mmol) and the mixture was stirred at 25 °C for 1 hour. Filtered and purified by preparative reversephase HPLC, affording the product (7.7mg, 14%).1H NMR (500 MHz, DMSO-d6) 5 8.52 (dd, J = 7.3, 1.2 Hz, 2H), 8.50 - 8.47 (m, 2H), 8.03 (t, J = 5.7 Hz, 1 H), 7.89 (dd, J = 8.2, 7.3 Hz, 2H), 4.26 (t, J = 6.5 Hz, 2H), 3.67 (t, J = 6.5 Hz, 2H), 3.61 - 3.55 (m, 2H), 3.52 - 3.47 (m, 2H), 3.36 (t, J = 5.8 Hz, 2H), 3.17 - 3.12 (m, 2H), 3.07 (d, J = 8.0 Hz, 2H). MS (ESI): m / z = 403.12 [M+H]+.Comparator Compound 10

[0177] To a solution of 2-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-1 H-benzo[de]isoquinoline-1 ,3(2H)-dione (60 mg, 1 Eq, 0.18 mmol) in DMF (2 mL) was added DIPEA (0.12 g, 0.16 mL, 5 Eq, 0.91 mmol) and di(1 H-imidazol-1 - yl)methanone (39 mg, 1 .3 Eq, 0.24 mmol), and the mixture was stirred at 25 °C for 30 min. Then hydroxylamine hydrochloride (25 mg, 2 Eq, 0.37 mmol) was added to the solution, and the mixture was stirred at 60 °C for 12 hours. Purified by pre-HPLC, affording the product (40.2mg, 57%).1H NMR (500 MHz, DMSO-d6) 5 8.57 (s, 1 H), 8.51 (dd, J = 7.3, 1 .2 Hz, 2H), 8.47 (dd, J = 8.3, 1 .1 Hz, 2H), 8.31 (s, 1 H), 7.88 (dd, J = 8.3, 7.2 Hz, 2H), 6.56 (t, J = 5.9 Hz, 1 H), 4.25 (t, J = 6.5 Hz, 2H), 3.66 (t, J = 6.5 Hz, 2H), 3.56 (dd, J = 5.9, 3.6 Hz, 2H), 3.50 - 3.44 (m, 2H), 3.36 (d, J = 6.1 Hz, 2H), 3.12 (q, J = 6.1 Hz, 2H). MS (ESI): m / z = 388.12 [M+H]+.

[0178] To a solution of 1 H,3H-benzo[de]isochromene-1 , 3-dione (300 mg, 1.0 Eq, 1.51 mmol) and 2-(2-(2- aminoethoxy)ethoxy)ethan-1-ol (271 mg, 1.2 Eq, 1.82 mmol) in DMF (2 mL) was stirred at 125 °C for 12 hours. Removed the solvent and purified with CombiFlash (MeOH / DCM 0-10%), affording the product (402mg, 80.6%).

[0179] To a solution of 2-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)-1 H-benzo[de]isoquinoline-1 ,3(2H)-dione (100 mg, 1 Eq, 304 pmol) in Pyridine (3 mL) was added phosphorothioyl trichloride (309 mg, 185 pL, 6 Eq, 1.82 mmol) at -15 °C, and the mixture was stirred at 0 °C for 30 min. Then the mixture was poured over a solution of NaHCOa (153 mg, 6 Eq, 1.82 mmol) in H2O (50 mL). The mixture was stirred at 0 °C for 2 hours. The mixture was evaporated to dryness under reduced pressure, and the residue was purified by pre-HPLC to afford the product (25.0mg, 19.4%).1H NMR (500 MHz, DMSO-d6) 5 12.01 (s, 2H), 8.51 (dd, J = 7.3, 1.2 Hz, 2H), 8.47 (dd, J = 8.4, 1 .2 Hz, 2H), 7.88 (dd, J = 8.2, 7.3 Hz, 2H), 4.25 (t, J = 6.5 Hz, 2H), 3.92 - 3.85 (m, 2H), 3.67 (t, J = 6.5 Hz, 2H), 3.57 (dd, J = 5.9, 3.5 Hz, 2H), 3.54 - 3.43 (m, 4H). MS (ESI): m / z = 426.03 [M+H]+.

[0180] To a solution of 1 H,3H-benzo[de]isochromene-1 , 3-dione (300 mg, 1 Eq, 1.51 mmol) in DMF (5 mL) was added 2-(2-aminoethoxy)ethan-1-ol (191 mg, 1.2 Eq, 1.82 mmol) and the mixture was stirred at 125 °C for 12 hour. Removed the solvent and purified by ISCO (MeOH / DCM 0-5%) to afford the product (400mg, 92.6%).

[0181] To a solution of 2-(2-(2-hydroxyethoxy)ethyl)-1 H-benzo[de]isoquinoline-1 ,3(2H)-dione (100 mg, 1 Eq, 351 pmol) diethyl vinylphosphonate (57.5 mg, 1 Eq, 351 pmol)in dry DMF (3 mL) was added K2CO3 (96.7 mg, 2 Eq, 701 pmol) and the mixture was stirred at 80 °C for 24 hour. Filtered, removed the solvent, and purified by ISCO (MeOH / DCM 0-5%) to give the product (50mg, 32%).

[0182] To a solution of diethyl (2-(2-(2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)- yl)ethoxy)ethoxy)ethyl)phosphonate (50 mg, 1 Eq, 0.11 mmol) in DCM (3 mL) was added bromotrimethylsilane (0.34 g, 0.29 mL, 20 Eq, 2.2 mmol) and the mixture was stirred at 25 °C for 8 hour. Evaporated to dryness under reduced pressure and purified with pre-HPLC gave the product (9.9mg, 23%).1H NMR (500 MHz, DMSO-rte) 5 8.51 (dd, J = 7.2, 1 .2 Hz, 2H), 8.47 (dd, J = 8.3, 1 .1 Hz, 2H), 7.88 (dd, J = 8.2, 7.3 Hz, 2H), 4.24 (t, J = 6.5 Hz, 2H), 3.66 (t, J = 6.6 Hz, 2H), 3.58 - 3.54 (m, 2H), 3.51 (dt, J = 9.3, 7.3 Hz, 2H), 3.47 - 3.43 (m, 2H), 1 .84 - 1 .73 (m, 2H). MS (ESI): m / z = 394.02 [M+H]+.Comparator Compound 13 (Compound 32)

[0183] To a solution of 1 H-benzo[de]isoquinoline-1 ,3(2H)-dione (200 mg, 1 Eq, 1.01 mmol) and tert-butyl 2- bromoacetate (198 mg, 1 Eq, 1.01 mmol) in DMF (5 mL) was added K2CO3 (140 mg, 1 Eq, 1.01 mmol) and the mixture was stirred at 50 °C for 5 hours. Filtered and remove the solvent. Then purified with CombiFlash (MeOH / DCM 0 to 10%), affording the desired product (136 mg, 43.1 %). MS (ESI): m / z = 339.1 [M+Na]+.

[0184] To a solution of tert-butyl 2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl) acetate (40 mg, 1 Eq, 0.13 mmol) in DCM (2 mL) was added TFA (1 mL), and the mixture was stirred at 25 °C for 30 min. Then remove the solvents and put it into the next step directly.

[0185] To a solution of the above 2-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl) acetic acid and HATU (73 mg, 1.5 Eq, 0.19 mmol) in DMF (2 mL) was added DIPEA (50 mg, 3 Eq, 0.39 mmol) and O-(tetrahydro-2H-pyran- 2-yl) hydroxylamine (15 mg, 1 Eq, 0.13 mmol)and the mixture was stirred at 25 °C for 30 min. NaHCOa aq.solution work up, extracted with CHCla / iPrOH (4: 1), dried over anhydrous Na2SO4, remove solvents and then purify with CombiFlash (MeOH / DCM 0 to 5%) affording product (24 mg, 52.1%).

[0186] To a solution of 2-(1 ,3-dioxo-l H-benzo[de]isoquinolin-2(3H)-yl)-N-((tetrahydro-2H-pyran-2- yl)oxy)acetamide (24 mg, 1 Eq, 0.068 mmol) in MeOH (2 mL) and Water (1 mL) was added TFA (0.2 mL) and the mixture was stirred at 25 °C for 2 hour. Purification by preparative reverse-phase HPLC (5- 95% MeOH in water, 25min, 20 mL / min) afforded the product compound Comp. 13 (11 mg, 60.0%) as a white powder.1H NMR (500 MHz, DMSO-d6) 5 10.80 (s, 1 H), 8.91 (s, 1 H), 8.52 (ddd, J = 8.3, 4.9, 1.2 Hz, 4H), 7.95 - 7.87 (m, 2H), 4.61 (s, 2H). MS (ESI): m / z = 293.07 [M+Na]+.Comparator Compound 14

[0187] To a solution of compound 1 (970 mg, 4.90 mmol, 1.0 eq) in ethanol (20 mL) was added compound 2 (3.00 g, 14.7 mmol, 3.0 eq), and the resulting mixture was stirred at 80 °C for 1 hour. On completion, the mixture was cooled to rt, diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were concentrated and the crude product was purified by flash column chromatography (PE / EA, 10 / 1) to afford compound 3 (1 .70 g, 90%) as yellow oil. LCMS: 385.20 [M+H]+

[0188] To a solution of compound 3 (1.70 g, 6.63 mmol, 1.0 eq) in dioxane (4 mL) was added HCI solution (4 M in dioxane, 50 mL). Then the solution was stirred at rt for 1 hour. The reaction was completed (detected by TLC and LCMS). The reaction solution was concentrated to afford compound 4 (1 .5 g, crude) as yellow oil, which was used in the next step without further purification. LCMS: 285.15 [M+H]+

[0189] To a solution of compound 4 (1.5 g, crude), compound 5 (563 mg, 3.52 mmol), DIEA (1.36 mg, 10.5 mmol) in DMF (20 mL) was added HATU (2.04 g, 5.37 mmol). The reaction mixture was stirred at room temperature for 3 hours. LCMS and TLC showed the completion of the reaction. The reaction was quenched with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layer was concentrated and purified by prep-TLC to afford compound 6 (1.70 g, 60% over two steps) as a yellow oil. LCMS: 427.20 [M+H]+

[0190] To a solution of compound 6 (1 .70 g, 3.99 mmol, 1 .0 eq) in dioxane (5 mL) was added HCI solution (4 M in dioxane, 20 mL) and the resulting mixture was stirred at rt for 1 hour. TLC showed compound 6 disappeared. The reaction mixture was concentrated to afford compound 7 (1 .47 g, 99%) as a yellow solid. LCMS: 371.20 [M+H]+

[0191] To a solution of compound 7 (500 mg, 1 .35 mmol, 1 .0 eq), TCFH (378 mg, 1 .35 mmol, 1 .0 eq), NMI (221 mmol, 2.70 mmol, 2.0 eq) in acetonitrile (10 mL) was added NH2OTHP (8, 247 mg, 2.11 mmol, 1.2 eq). The reaction mixture was stirred at room temperature for 3 hours. LCMS and TLC showed the completion of the reaction. The reaction was quenched with water (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layer was concentrated and purified by prep-TLC to afford compound 9 (600 mg, 95%) as a yellow oil. LCMS:470.20 [M+H]+

[0192] To a solution of the compound 9 (230 mg, 0.490 mmol, 1.0 eq) in ethanol (3mL) was added TsOH^O (17 mg, 0.089 mmol, 0.2 eq). The reaction mixture was allowed to stir at room temperature overnight, during which, a tan precipitate formed in solution. The solid was purified by prep-HPLC (5% to 95% ACN in water) to afford Comp. 14 (102 mg, 54%) as white solid. LCMS:385.90 [M+H]+ 1H NMR (400 MHz, DMSO-d6) 5 10.43 (brs, 1 H), 8.85 (s, 1 H), 8.49 (d, J = 8.0 Hz, 2H), 8.45 (d, J = 8.0 Hz, 2H), 7.99 - 7.95 (m, 1 H), 7.86 (t, J = 8.0 Hz, 2H), 4.23 (t, J = 7.6 Hz, 2H), 3.64 (t, J = 7.6 Hz, 2H), 3.45 (t, J = 7.6 Hz, 2H), 3.20 - 3.14 (m, 2H), 2.85 (s, 2H).Comparator Compound 15

[0193] To a solution of compound 1 (5.0 g, 24.4 mmol, 1 .0 eq) in toluene (50 mL) were added compound 2 (10.4 g, 73.2 mmol, 3.0 eq) and CS2CO3 (15.9 g, 48.8 mmol, 2.0 eq). The reaction mixture was stirred at 50 °C for 16 hours. LCMS and TLC showed the formation of compound 3. The mixture was filtered and the filtrate was concentrated. The crude product was purified by flash column chromatography (PE / EA, 3 / 1) to afford compound 3 (2.3 g, 27%) as colorless oil. LCMS: 348.05 [M+H]+

[0194] To a solution of compound 3 (2.30 g, 6.63 mmol, 1 .0 eq) in dioxane (4 mL) was added HCI solution (4 M in dioxane, 15 mL). Then the solution was stirred at rt for 1 hour. The reaction was completed (detected byTLC and LCMS). The reaction solution was concentrated to afford compound 4 (1 .4 g, crude) as yellow oil, which was used in the next step without further purification. LCMS: 191.90 [M+H]+

[0195] To a solution of compound 4 (1 .4 g, crude) in DMF (20 mL) were added DIEA (709 mg, 5.50 mmol), compound 5 (724 mg, 3.66 mmol). The reaction mixture was heated at 90 °Cfor 2 hours. The reaction solution was partitioned between water (50 mL) and EA (100 mL x 3). The organic layer was concentrated and purified by flash column chromatography (PE / EA = 1 / 1) to afford compound 6 (500 mg, 20% over two steps) as yellow oil. LCMS:372.90 [M+H]+

[0196] To a solution of compound 6 (500 mg, 1.35 mmol, 1.0 eq) in acetonitrile (10 mL) were added N,N,N',N'- tetramethylchloroformamidinium hexafluorophosphate (TCFH, 415 mg, 1.48 mmol, 1.1 eq), N-methylimidazole (NMI, 332 mg, 4.04 mmol, 3.0 eq), compound 7 (260 mg, 2.22 mmol, 1.5 eq). The mixture was stirred at room temperature for 16 h. The reaction was completed (detected by TLC). The mixture was diluted with water (10 mL) and extracted with ethyl acetate (25 mL x 2). The combined organic layers were concentrated and purified by flash column chromatography (PE / EA, 3 / 1) to afford compound 8 (230 mg, 36%) as white solid. LCMS: 471.50 [M+H]+

[0197] To a solution of the compound 8 (230 mg, 0.488 mmol, 1.0 eq) in ethanol (3 mL) was added TsOH'hhO (17 mg, 0.089 mmol, 0.2 eq). The reaction mixture was allowed to stir at room temperature for 16 hours, during which, a tan precipitate formed in solution. The precipitate was purified by prep-HPLC (5% to 95% ACN in water) to afford Comp. 15 (144.8 mg, 77%) as light yellow sticky solid. LCMS: 387.00 [M+H]+HNMR: (400 MHz, DMSO-cfe) 5 10.34 (s, 1 H), 8.68 (s, 1 H), 8.49 (d, J= 8.0 Hz, 2H), 8.46 (d, J = 8.0 Hz, 2H), 7.86 (t, J = 8.0 Hz, 2H), 4.26 - 4.19 (m, 2H), 3.63 (t, J= 7.6 Hz, 2H), 3.53 - 3.48 (m, 2H), 3.42 - 3.39 (m, 2H), 3.38 - 3.35 (m, 1 H), 3.21 - 3.15 (m, 1 H), 2.31 - 2.23 (m, 1 H), 0.81 (d, J= 7.2 Hz, 3H).Comparator Compound 16 (Compound 33)

[0198] To a solution of 1 H,3H-benzo[de]isochromene-1, 3-dione (100 mg, 1 Eq, 505 pmol) tert-butyl 3- aminopropanoate (73.3 mg, 1 Eq, 505 pmol) in DMF (4 mL) was stirred at 125 °C for 8 hours. Removed thesolvent and purified with CombiFlash (MeOH / DCM 0 to 10%) affording the desired product (100 mg, 60.9%). MS (ESI): m / z = 348.1 [M+Na]+.

[0199] To a solution of tert-butyl 3-(1 ,3-dioxo-1 H-benzo[de]isoquinolin-2(3H)-yl) propanoate (30 mg, 1 Eq, 92 pmol) in DCM (2 mL) was added TFA (1 mL) and the mixture was stirred at 25 °C for 30 min. Then remove the solvents and put them into the next step directly.

[0200] To a solution of above acid and O-(tetrahydro-2H-pyran-2-yl) hydroxylamine (13 mg, 1.2 Eq, 0.11 mmol) in DMF (2 mL) was added HATU (70 mg, 2 Eq, 0.18 mmol) DIPEA (36 mg, 48 pL, 3 Eq, 0.28 mmol) and the mixture was stirred at 25 °C for 30 min. NaHCOa aq. solution work up, extracted with CHCla / iPrOH (4:1), dried over anhydrous Na2SO4, remove solvents and then purify with CombiFlash (MeOH / DCM 0 to 5%) affording product (19 mg, 56.1 %).

[0201] To a solution of the above product in MeOH (4 mL) and H2O (1 mL) was added TFA (0.5 mL). Then stirred at RT for 2h. Purification by preparative reverse-phase HPLC (1- 95% MeOH in water, 25min, 20 mL / min) afforded the product compound Comp. 16 (9 mg, 30.0%) as a white powder.1H NMR (500 MHz, DMSO-cfe) 3 10.50 (s, 1 H), 8.51 (dd, J = 7.3, 1.1 Hz, 2H), 8.48 (dd, J = 8.3, 1.1 Hz, 2H), 7.89 (dd, J = 8.2, 7.3 Hz, 2H), 4.30 - 4.24 (m, 2H), 2.41 - 2.34 (m, 2H). MS (ESI): m / z = 285.09 [M+H]+.Comparator Compound 17 (Compound 34)

[0202] The title compound was prepared according to the same procedure as Comp. 13. Purification by preparative reverse-phase HPLC (1- 95% MeOH in water, 25min, 20 mL / min) afforded the product as a white powder (13 mg, 4 steps yield 29%).1H NMR (500 MHz, DMSO) 5 10.28 (s, 1 H), 8.44 (dd, J = 7.3, 1.2 Hz, 2H), 8.40 (dd, J = 8.3, 1 .1 Hz, 2H), 7.83 - 7.79 (m, 2H), 4.00 (t, J = 7.2 Hz, 2H), 1 .98 (dd, J = 8.9, 6.6 Hz, 2H), 1 .85 - 1.75 (m, 2H). MS (ESI): m / z = 299.02 [M+H]+.Comparator Compound 18 (Compound 35)

[0203] The title compound was prepared according to the same procedure as Comp. 13. Purification by preparative reverse-phase HPLC (1- 95% MeOH in water, 25min, 20 mL / min) afforded the product as a white powder (13 mg, 4 steps yield 18%).1H NMR (500 MHz, DMSO-d6) 6 10.33 (d, J = 1.7 Hz, 1 H), 8.65 (d, J = 1.8 Hz, 1 H), 8.50 (dd, J = 7.2, 1.1 Hz, 2H), 8.47 (dd, J = 8.4, 1.2 Hz, 2H), 7.88 (dd, J = 8.2, 7.2 Hz, 2H), 4.05 (t, J = 7.0 Hz, 2H), 1.99 (t, J = 7.1 Hz, 2H), 1.67 - 1.50 (m, 4H). MS (ESI): m / z = 313.02 [M+H]+.Comparator Compound 19 (Compound 37)

[0204] The title compound was prepared according to the same procedure as compound Comp. 13.Purification by preparative reverse-phase HPLC (5- 95% MeOH in water, 25min, 20 mL / min) afforded the product as a white powder (12 mg, 4 steps yield 13%).1H NMR (500 MHz, DMSO-d6) 5 10.32 (s, 1 H), 8.50 (dd, J = 7.3, 1 .2 Hz, 2H), 8.46 (dd, J = 8.4, 1 .1 Hz, 2H), 7.88 (dd, J = 8.2, 7.3 Hz, 2H), 4.06 - 4.02 (m, 2H), 1 .94 (t, J = 7.4 Hz, 2H), 1 .62 (p, J = 7.3 Hz, 2H), 1 .49 (p, J = 7.3 Hz, 2H), 1 .38 - 1 .25 (m, 4H). MS (ESI): m / z = 341 .02 [M+H]+.Comparator Compound 20 (Compound 38)

[0205] The title compound was prepared according to the same procedure as Comp. 16. Purification by preparative reverse-phase HPLC (5- 90% MeOH in water, 25min, 20 mL / min) afforded the product as a white powder (14 mg, 4 steps yield 22%).1H NMR (500 MHz, DMSO-d6) 5 10.31 (d, J = 1.6 Hz, 1 H), 8.63 (d, J = 1.8 Hz, 1 H), 8.50 (dd, J = 7.3, 1.1 Hz, 2H), 8.46 (dd, J = 8.3, 1.1 Hz, 2H), 7.87 (dd, J = 8.2, 7.2 Hz, 2H), 4.06 - 4.01 (m, 2H), 1 .92 (t, J = 7.4 Hz, 2H), 1 .63 (t, J = 7.3 Hz, 2H), 1 .52 - 1 .43 (m, 2H), 1 .35 - 1.27 (m, 4H), 1 .27 - 1 .21 (m, 2H). MS (ESI): m / z = 354.95 [M+H]+.Comparator Compound 21 (Compound 39)

[0206] The title compound was prepared according to the same procedure as Comp. 16. Purification by preparative reverse-phase HPLC (5- 85% MeOH in water, 25min, 20 mL / min) afforded the product as white powder (41 mg, 4 steps yield 24%).1H NMR (500 MHz, DMSO-d6) 5 10.30 (d, J = 1.6 Hz, 1 H), 8.63 (d, J = 1.8 Hz, 1 H), 8.50 (dd, J = 7.2, 1.2 Hz, 2H), 8.46 (dd, J = 8.3, 1.1 Hz, 2H), 7.87 (dd, J = 8.2, 7.3 Hz, 2H), 4.07 - 4.00 (m, 2H), 1 .92 (t, J = 7.4 Hz, 2H), 1 .63 (p, J = 7.3 Hz, 2H), 1 .47 (p, J = 7.3 Hz, 2H), 1 .37 - 1 .29 (m, 4H), 1 .29 - 1.19 (m, 4H). MS (ESI): m / z = 368.95 [M+H]+.Comparator Compound 22

[0207] To a solution of 2-(2-(2-azidoethoxy)ethoxy)ethane-1-sulfonic acid (100 mg, 1 Eq, 418 pmol) in MeOH (4 mL) was added Pd / C (30 mg, 1 Eq, 418 pmol). Exchanged with H2 three times, and the mixture was stirred at 25 °C for 30 min. Filtered, removed the solvent, and put it into the next step directly.

[0208] To a solution of 2-(2-(2-aminoethoxy)ethoxy)ethane-1-sulfonic acid (60 mg, 1 Eq, 0.28 mmol) in DMF (4 mL) was added 1 H,3H-benzo[de]isochromene-1 , 3-dione (67 mg, 1.2 Eq, 0.34 mmol) and the mixture was stirred at 125 °C for 12 hour. Purified by pre-HPLC (49mg, 44%).1H NMR (500 MHz, DMSO-d6) 3 8.52 (dd, J = 7.3, 1.2 Hz, 2H), 8.47 (dd, J = 8.3, 1.1 Hz, 2H), 7.88 (dd, J = 8.2, 7.2 Hz, 2H), 4.24 (t, J = 6.6 Hz, 2H), 3.65 (t, J = 6.5 Hz, 2H), 3.60 - 3.50 (m, 4H), 3.48 - 3.41 (m, 2H), 2.68 - 2.61 (m, 2H). MS (ESI): m / z = 392.07 [M-H]-.Biological Assays

[0209] Mouse PTER protein purification

[0210] The mouse PTER (mPTER) gene (Uniprot accession number Q60866) was codon-optimized for bacterial expression and synthesized as gBIocks by IDT. The gene fragment was cloned into the pET-20b vector with a C-terminal hexa-Histidine (His) tag and a N-terminal Strep tag. The pET-20b-mPTER plasmid was transformed into BI21 competent cells (ThermoScientific, EC0114) and grown in LB medium supplemented with ampicillin at 37 °C with shaking overnight.

[0211] For protein expression, BL21 cells were inoculated into autoinduction medium containing the following components: 10 g tryptone (FisherScientific, BP1421 -500), 5 g yeast extract (FisherScientific, BP1422-500), 2 ml MgSC>4 (1 M), 1 ml metal solution (0.05 M Feccir citrate, 0.02 M CaCl2, 0.02 M ZnSC>4, 2 piM C0CI2, 2 piM CUSO4, 2 piM NiCh, 2 M Na2MoC>4, 2 pM Boric acid), 20 ml salt solution (167.5g Na2HPC>4, 85g KH2PO4, 53.4g NH4CI and 17.8g Na2SC>4 in 500 ml water in total) and 20 ml sugar solution (125 g glycerol, 12.5 g glucose and 50 g a-lactose in 500 ml water in total) to a final volume of 1 L. Cells were grown to optical density (CD600) of 0.5-0.7 at 37 °C, then incubated at15 °C overnight. Cells were harvested via centrifugation at 8,000 rpm for 30 min at 4 °C and lysed in buffer (50 mM Tris-HCI, 135mM NaCI, pH7.4) by probe sonication on ice. Soluble fractions were obtained by centrifugation at 15,000 rpm for 30 min at 4 °C. The recombinant mPTER proteins were purified using Strep-Tactin resins (IBA, 2-1208-002), according to the manufacturer's instructions. Bound mPTER proteins were eluted with buffer (2.5 mM D-Desthiobiotin, 25 mM Tris and 130 mM NaCI pH7.4). Purified proteins were aliquoted and stored at -80 °C for subsequent enzymatic assays.

[0212] In vitro N-acetyl taurine hydrolysis assays

[0213] Enzymatic assays were performed using 200 ng of recombinant mPTER proteins incubated with 100uM N-acetyltaurine (Cayman, 35169) and the compound in a 50 pil reaction buffer (50 mM Tris-HCI, 135mM NaCI, pH7.4) at 37 °C for 1 hour. The reactions were then quenched by the addition of 150 pil of a 2:1 (v / v) mixture of acetonitrile and methanol. The mixture was centrifuged at 15,000 rpm for 30 min at 4 °C to remove precipitated proteins.

[0214] For enzymatic assays performed in a 96-well plate format, the quenched reaction mixture was first centrifuged at 2000 rpm for 1 min. The supernatant was then transferred to a 96-well PVDF membrane filter and centrifuged at 2,000 rpm for 2 min at room temperature. Then filtrate was then transferred to mass spec vials or a 96-well plate compatible with the LC-MS system for subsequent metabolite analysis. To determine the half- maximal inhibitory concentration (IC50) values, compounds were titrated from 1 nM to 100 pi M using a 6-point concentration range. The IC50 values were calculated by fitting the dose-response data to a four-parameter logistic curve using GraphPad Prism software (version 9.0, GraphPad Software, Inc.).

[0215] In vitro HDAC activity assays

[0216] The liver tissues were homogenized in HDAC reaction assay buffer (HEPES 50mM, NaC1 137 mM, KCI 3mM, MgCl2 1mM, pH8.0) at a ratio of 100 mg tissue per ml of buffer, and further sonicated. The lysates were centrifuged at 15,000 rpm for 30 min at 4 °C. The supernatant protein concentration was determined using a Nanodrop One. The HDAC assay was performed in 50 pil reaction volume containing the HDAC reaction assay buffer, 600 ug of the liver lysate, 100 piM HDAC substrate peptide Ac-Arg-Gly-Lys-Ac-Glu-AMC (custom synthesized by Elim Biopharm), and 10 piM of compound. The reaction mixture was incubated at 37 °C for 1 hour, then quenched by the addition of 150 pil of a 2:1 (v / v) mixture of acetonitrile and methanol. The mixture was centrifuged at 15,000 rpm for 30 min at 4 °C to remove precipitated proteins. For enzymatic assays performed in a 96-well plate format, the quenched reaction mixture was first centrifuged at 2000 rpm for 1 min. The supernatant was then transferred to a 96-well PVDF membrane filter and centrifuged at 2,000 rpm for 2 min at room temperature. Then filtrate was then transferred to mass spec vials or a 96-well plate compatible with the LC-MS system for subsequent metabolite analysis. The IC50 values were calculated by fitting the dose-response data to a four-parameter logistic curve using GraphPad Prism software (version 9.0, GraphPad Software, Inc.). Results of PTER and HDAC inhibition for known HDAC inhibitors Vorinostat, Droxinostat and Scriptaid are shown in Figure 1 . Results of PTER and HDAC inhibition for Compounds 32-42 are shown in Figure 2a.Compounds 32-39 structures areCompound32 is also referred to as Comparator Compound 13; Compound 33 as Comparator Compound 16; Compound 34 as Comparator Compound 17; Compound 35 as Comparator Compound 18; Compound 37 as Comparator Compound 19; Compound 38 as Comparator Compound 20; and Compound 39 as Comparator Compound 21. A graph representation of the PTER vs. HDAC inhibition activity of Compound 41 is also shown in Figure 2b.

[0217] IC 50 determination for Compound 41 against all HDAC family members (HDAC 1-11 and SIRT 1, SIRT 2, SIRT 3, SIRT 5) were determined by Reaction Biology Corporation using their standard assay protocols. The results are shown in Figures 3 and 4.

[0218] Sample preparation and western blotting

[0219] For cell culture samples, cells were harvested and lysed by probe sonication, boiled for 10 min at 95 °C in 4x NuPAGE LDS Sample Buffer (ThermoFisher, NP0008) supplemented with 100 mM DTT (Sigma, D0632- 1G). Proteins were separated on Novex™ Tris-Glycine Mini Protein Gels 4-20 % (Invitrogen, LC2675) and transferred to nitrocellulose membranes. Equal loading was confirmed by Ponceau S staining. Membranes were blocked with Odyssey blocking buffer for 30 min at room temperature and incubated overnight at 4 °C with primary antibodies, 1 :5000 dilution rabbit anti-p-actin antibody (Abeam, ab8227), 1 :1000 recombinant Anti- Histone H4 (acetyl K16) antibody (Abeam, ab109463), 1 :1000 Histone H3K27ac antibody (active motif, 39134). Membranes were washed three times with PBST (0.05% Tween-20 in PBS) and stained with species-matched secondary antibodies (1 :10000 dilution goat anti-rabbit IRDye 800RD (LI-COR, 925-68070) and 1 :10000 dilution goat anti-mouse IRDye 680RD (LI-COR, 925-68070)) at room temperature for 1 h. Blots were further washed three times with PBST and imaged with the Odyssey CLx Imaging System The results showed that treatment of cells with compound 41 had no effect on histone H3 and histone H4 acetylation, suggesting compound 41 does not affect the HDACs responsible for the deacetylation of histone H3 and histone H4.

[0220] Preparation of mouse blood for LC-MS analysis

[0221] Mouse blood was collected and centrifuged at 5000 rpm for 5 min at 4 °C to obtain plasma. A 50 l aliquot of plasma was mixed with 150 pl of a 2:1 (v / v) mixture of acetonitrile and methanol and vortexed for 30 s to precipitate proteins. The mixture was then centrifuged at 15,000 rpm for 30 min at 4 °C and the supernatant was transferred to a LC-MS vial for subsequent analysis. For time course experiments, approximately 100 pl of blood was collected at each time point. A 20 pl aliquot of plasma was used for metabolite isolation, and the volume of the acetonitrile: methanol mixture was proportionally reduced to maintain the 1 :3 (plasma:solvent) ratio.

[0222] As seen in Figure 5, showing the increase in N-acetyltaurine after administration of Compound 41, inhibition of PTER by Compound 41 resulted in a dose dependent increase in N-acetyltaurine levels in lean mice as well as increases in N-acetyltaurine levels up to 1 -hour post-injection in lean mice and up to 2-hours postinjection in obese mice.

[0223] Measurement of N-acetyltaurine by LC-MS

[0224] Measurements were performed using an Agilent 6545 Quadrupole time-of-flight LC-MS instrument as previously described. MS analysis was conducted using electrospray ionization (ESI) in negative mode. The dual ESI source parameters were configured as follows: the gas temperature, 250 °C; drying gas flow, 12 l / min; nebulizer pressure, 20 psi; capillary voltage, 3,500 V; and the fragmentor voltage, 100 V. The separation of polar metabolites was achieved using a Luna 5 pm NH2 100 A LC column (Phenomenex 00B-4378-E0) with normal phase chromatography. Mobile phases were as follows: buffer A, 95:5 water: acetonitri le with 0.2% ammonium hydroxide and 10 mM ammonium acetate; buffer B, acetonitrile. For the in vitro assay, the LC gradient initiated at 100% B with a flow rate of 0.7 ml / min from 0 to 0.2 min. The gradient was then linearly increased to 20% A / 80% B at a flow rate of 0.7 ml / min from 0.4 to 1 .1 min. From 1 .1 to 4 min, the gradient was linearly increased to 50% A / 50% B at the same flow rate. The gradient was maintained at 50% A / 50% B at a flow rate of 0.7 ml / min from 4min to 4.5min. The gradient was decreased to 0%A / 100%B from 4.5min to 5min and maintained at 0%A / 100%B from 5min to 6min. N-acetyltaurine (Cayman, 35169) eluted around 4 min and taurine (sigma, T0625-500G) eluted around 4.1 min under the above conditions.

[0225] For in vivo assay samples, the LC gradient initiated at 100% B with a flow rate of 0.7 ml / min from 0 to 2 min. The gradient was then linearly increased to 50% A / 50% B at a flow rate of 0.7 ml / min from 2 to 20 min. From 20 to 25 min, the gradient was maintained at 50% A / 50% B at a flow rate of 0.7 ml / min. The gradient was maintained at 0%A / 100%B from 25min to 30min. N-acetyltaurine (Cayman, 35169) eluted around 11.5 min and taurine (sigma, T0625-500G) eluted around 12.8 min under the above conditions.

[0226] General animal information

[0227] All animal experiments were conducted in accordance with protocols approved by the Stanford University Administrative Panel on Laboratory Animal Care. Mice were maintained in a facility with 12-h light-dark cycles at 22 °C and approximately 50% relative humidity. The mice were fed a standard irradiated rodent chow diet, except where indicated, in which case a high-fat diet (D12492, Research Diets 60% kcal from fat) was used. Male C57BL / 6 J (stock number 000664) and male C57BL / 6 J DIO mice (stock number 380050) were purchased from the Jackson Laboratory. For intraperitoneal injections performed for the data as shown for Figure 5, Compound 41 was dissolved in an 18:1 :1 mixture of saline:DMSO:kolliphor. For intraperitoneal injections performed for the data in Figure 6, Compound 41 was dissolved in a 1 :1 mixture of DMSO:kolliphor.

[0228] Food intake studies

[0229] Mice were individually housed and weights of food at time 0 were taken. Mice were injected with either vehicle or compound 41 (100 mg / kg) at 6 PM. Food intake was measured at 6 and 18 hours. Results are shown in Figure 6. Figure 7 shows differences in body weight of mice that were injected with either vehicle or compound 41 (100 mg / kg) relative to cumulative food intake over the course of 8 days.

[0230] Mice were also individually housed and injected with either vehicle or Compound 41 (100 mg / kg) for 3 weeks to evaluate the effect of compound 41 on cumulative food intake. Results from this study are shown in Figures 8-10. Figure 8 shows the cumulative food intake of mice injected with either vehicle or compound 41 (100 mg / kg) relative to cumulative food intake over the course of 24 hours. Figure 9 shows the respiratory exchange ratio of the mice over the course of 24 hours. Figure 10 shows the activity of mice injected with either vehicle or compound 41 over the course of 24 hours.

[0231] Another group of DIO mice were injected with either vehicle or compound 41 (100 mg / kg) over Figures 11 and 12 show the cumulative food intake relative to cumulative food intake over the course of 27 days and the change in body weight over the course of 27 days of mice injected with either vehicle or compound 41 (100 mg / kg), respectively. Dissection of tissues at the end of the study showed a loss of adipose mass with no changes in the masses of the other organs, including liver, kidney, and heart (Data not shown).

[0232] Combination therapy studies

[0233] DIO mice were treated daily with either the GLP-1R agonist Semaglutide alone (0.3 nmol / kg / day, IP) or a combination of Semaglutide and compound 41 (0.3 nmol / kg / day Semaglutide, IP, and 100 mg / kg / day compound 41, IP) to determine the efficacy of compound 41 when used directly in combination with GLP-1R agonists. Figures 13 and 14 show the change in body weight and cumulative food intake over a one week treatment period for mice treated with vehicle, Semaglutide alone, and a combination of Semaglutide and compound 41. Semaglutide-treated mice ate less than vehicle-treated mice, resulting in weight loss compared to vehicle-treated mice (Figures 13, 14). In the Semaglutide and compound 41 combination group, cumulative food intake was further reduced, and body weight was correspondingly further lowered compared to vehicle-treated mice (P < 0.001).

[0234] Next, the impact of compound 41 in a mouse model of incretin discontinuation-associated weight rebound was assessed. The prevalence of drug discontinuation in humans has been estimated to be -30-50%within 12 months of GLP-1RA initiation. Weight regain is commonly observed after drug discontinuation. To model this phenomenon, DIO mice were treated with Semaglutide (2 nmol / kg / day, IP) for 15 days (weight loss phase) and then switched to vehicle treatment for 10 days (weight regain phase). The experimental group consisted of DIO mice treated with Semaglutide (2 nmol / kg / day, IP) during the weight loss phase and then switched to compound 41 (100 mg / kg / day, IP) during the weight regain phase. As additional comparators, control groups of DIO mice were maintained on only vehicle or only Semaglutide (2 nmol / kg / day, IP) for the entire treatment protocol. During the initial treatment phase, all mice lost body weight (mean ± SEM, -4.1 g ± 0.4 g) whereas vehicle-treated mice gained weight (mean ± SEM, +2.7 g ± 0.4 g, P < 0.001), resulting in a -16% net vehicle-adjusted weight loss. In the weight regain phase, Semaglutide-treated mice that were switched to vehicle treatment rapidly gained weight (mean ± SEM, +3.1 g ± 0.5 g). By contrast, Semaglutide-treated mice that were then switched to compound 41 maintained their weight loss (mean ± SEM, -0.5 g ± 0.7 g, P < 0.001) at a level comparable with mice that continued Semaglutide therapy (mean ± SEM, -0.6 g ± 0.3 g). Thus, compound 41 is effective for weight maintenance and the prevention of weight regain following GLP-1RA discontinuation.

[0235] Biological Activity Assays

[0236] The inhibitory activity of additional PTER inhibitors was evaluated and these results are presented in Table 1.

[0237] Table 1.

[0238] Comparative Examples

[0239] An inactive analog of compound 41 (PTERamide - also referred to as Comparator Compound 1 herein) was synthesized via chemical replacement of the hydroxamate with a primary amide in order to evaluate the metabolic effects of treatment with compound 41 and the biochemical activity of Comparator Compound 1, (Figure 16). Using recombinant PTER protein and N-acetyltaurine hydrolysis activity assays, Comparator Compound 1 was observed to be completely ineffective in inhibiting PTER enzyme activity (Figure 17). Further, injection of Comparator Compound 1 (100 mg / kg, IP) did not reduce acute food intake (Figure 18) and did not lower body weight upon chronic administration (100 mg / kg / day, IP, Figure 19) as opposed to injection of compound 41 (100 mg / kg).

[0240] Additional analogs were prepared and the PTER inhibitory activity of these analogs were evaluated as comparators. These results are presented in Table 2.TABLE 2.REFERENCES1 Lourengo, R. & Camilo, M. E. Taurine: a conditionally essential amino acid in humans? An overview in health and disease. NutrHosp 17, 262-270 (2002).2 Ripps, H. & Shen, W. Review: taurine: a "very essential" amino acid. Mol Vis 18, 2673-2686 (2012).3 Lambert, I. H., Kristensen, D. M., Holm, J. B. & Mortensen, 0. H. Physiological role of taurine-from organism to organelle. Acta Physiol (Oxf) 213, 191-212 (2015). https: / / doi.org / 10.1111 / apha.123654 Stipanuk, M. H. Metabolism of sulfur-containing amino acids. Annu Rev Nutr , 179-209 (1986). https: / / doi.org / 10.1146 / annurev.nu.06.070186.0011435 Jacobsen, J. G. & Smith, L. H. Biochemistry and physiology of taurine and taurine derivatives. Physiol Rev 48, 424-511 (1968). https: / / doi.Org / 10.1152 / physrev.1968.48.2.424Shi, X., Yao, D. & Chen, C. Identification of N-acetyltaurine as a novel metabolite of ethanol through metabolomics-guided biochemical analysis. J Biol Chem 287, 6336-6349 (2012). https: / / doi.org / 10.1074 / jbc.M111.312199Miyazaki, T. et al. Increased N-Acetyltaurine in the Skeletal Muscle After Endurance Exercise in Rat.Adv Exp Med Biol 975 Pt 1, 403-411 (2017). https: / / doi.org / 10.1007 / 978-94-024-1079-2_33Meyre, D. et al. Genome-wide association study for early-onset and morbid adult obesity identifies three new risk loci in European populations. Nat Genet 41, 157-159 (2009). https: / / doi.org / 10.1038 / ng.301Jong, C. J., Sandal, P. & Schaffer, S. W. The Role of Taurine in Mitochondria Health: More Than Just an Antioxidant. Molecules 26 (2021). https: / / doi.org / 10.3390 / molecules26164913Jia, F. et al. Taurine is a potent activator of extrasynaptic GABA(A) receptors in the thalamus. J Neurosci 28, 106-115 (2008). https: / / doi.org / 10.1523 / JNEUROSCI.3996-07.2008Huxtable, R. J. 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FASEB J 20, 574-576 (2006). https: / / doi.org / 10.1096 / fj.05-5016fjeWaldron, M., Patterson, S. D., Tallent, J. & Jeffries, O. The Effects of an Oral Taurine Dose and Supplementation Period on Endurance Exercise Performance in Humans: A Meta-Analysis. Sports Med 48, 1247-1253 (2018). https: / / doi.org / 10.1007 / s40279-018-0896-2Singh, P. et al. Taurine deficiency as a driver of aging. Science 380, eabn9257 (2023). https: / / doi.org / 10.1126 / science.abn9257Park, E. et al. Cloning of murine cysteine sulfinic acid decarboxylase and its mRNA expression in murine tissues. Biochim Biophys Acta 1574, 403-406 (2002). https: / / doi.org / 10.1016 / s0167- 4781 (01)00364-5McCoy, J. G. et al. Structure and mechanism of mouse cysteine dioxygenase. Proc Natl Acad Sci U S A 103, 3084-3089 (2006). https: / / doi.org / 10.1073 / pnas.0509262103 Veeravalli, S. et al. Flavin-Containing Monooxygenase 1 Catalyzes the Production of Taurine from Hypotaurine. 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Claims

What is claimed is:1 . A compound having a structure of Formula (I):or a pharmaceutically acceptable salt thereof; wherein:L is Cs galkylene or Ca-galkenylene, wherein the alkylene or alkenylene backbone is interrupted with 1 , 2, or 3 heteroatoms independently selected from NR1, 0, and S; andR1is H or CH3; each R2is independently N(RN)2 or NO2; each RNis independently H or Ci-salkyl; and n Is O, 1 , or 2.

2. The compound or salt of claim 1 , wherein L is 6-10 atoms long.

3. The compound or salt of claim 2, wherein L is 8 atoms long.

4. The compound or salt of any one of claims 1 to 3, wherein the alkylene backbone is interrupted with 1 heteroatom.

5. The compound or salt of claim 4, wherein the heteroatom is 0.

6. The compound or salt of claim 4, wherein the heteroatom is NR1.

7. The compound or salt of any one of claims 1 to 3, wherein the alkylene backbone is interrupted with 2 heteroatoms.

8. The compound or salt of claim 7, wherein at least one heteroatom is 0.

9. The compound or salt of claim 8, wherein each heteroatom is 0.

10. The compound or salt of claim 7 or 8, wherein at least one heteroatom is NR1.

11. The compound or salt of claim 10, wherein each heteroatom is NR1.

12. The compound or salt of claim 10, wherein one heteroatom is 0 and the other heteroatom isNR1.

13. The compound or salt of any one of claims 1 to 3, wherein the alkylene backbone is interrupted with 3 heteroatoms.

14. The compound or salt of claim 13, wherein at least one heteroatom is 0.

15. The compound or salt of claim 14, wherein each heteroatom is 0.

16. The compound or salt of claim 13 or 14, wherein at least one heteroatom is NR1.

17. The compound or salt of claim 16, wherein each heteroatom is NR1.

18. The compound or salt of claim 13, wherein at least one heteroatom is 0 and at least one heteroatom is NR1.

19. The compound or salt of any one of claims 1 to 4, 7, 8, 10, 13, 14, 16, and 18, wherein at least one heteroatom is S.

20. The compound or salt of any one of claims 1 to 4, 6 to 8, 10 to 14, and 16 to 19, wherein R1isCH3.

21. The compound or salt of any one of claims 1 to 4, 6 to 8, 10 to 14, and 16 to 19, wherein R1is H.

22. The compound or salt of claim 1 selected from23. A compound or pharmaceutically acceptable salt thereof having a structure as recited in TableA.

24. A pharmaceutical formulation comprising the compound or salt of any one of claims 1 to 23 and a pharmaceutically acceptable excipient.

25. A method of inhibiting PTER activity in a cell comprising contacting the cell with the compound or salt of any one of claims 1 to 23 in an amount effective to inhibit PTER activity.

26. A method of treating a disease or disorder associated with aberrant PTER activity in a subject, comprising administering to the subject a therapeutically effective amount of the compound or salt of any one of claims 1 to 23.

27. The method of claim 25, wherein the disease or disorder is obesity, cirrhosis, myocardial infarction, or alcoholism.

28. The method of claim 24 or 25, further comprising administering an additional therapeutic agent.

29. The method of claim 27, wherein the additional therapeutic agent is a GLP-1 R agonist.

30. The method of claim 28 or 29, wherein the GLP-1 R agonist is semaglutide.31 . A method of treating weight gain associated with discontinuation of a GLP-1 R agonist in a subject, comprising administering to the subject a therapeutically effective amount of the compound or salt of any one of claims 1 to 23.

32. The method of claim 31 , wherein the discontinued GLP-1 R agonist is semaglutide.

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