IL250730A

Inhibitors of -amino- -carboxymuconic acid semialdehyde decarboxylase

Abstract

This record has no abstract on file.

IL250730A, drawing sheet 1 — Sheet 1 of 170

Term

No projected expiry on record.

44 claims: 35 independent, 9 dependent

1
CLAIMS 1. A compound represented by Formula (I):X R= JL .R f Ϊ1 / r1 R d N C (I) or a pharmaceutically acceptable salt or tautomer thereof, wherein: X is O, OH, or Cl;L is -(CH 2 ) m CH 2 CH 2 -, -(CH 2 ) m Y(CH 2 ) p -, -(CH 2 ) m C(O)(CH 2 ) p -, -(CH 2 ) m C(O)O(CH 2 ) p -, -(CH 2 ) m C(O)NR 2 (CH2)p-, or -(CH2)mNR 2 C(O)(CH2) p ;Y is Ο, N or S(O) q ;R 1 is C6-C10 aryl or heteroaryl, wherein the aryl and heteroaryl are substituted with R a and R b , and optionally substituted with one or more R e ;R 2 is H or C1-C6 alkyl;one of R a and R b is hydrogen and the other is -(CH2)rCO2R x , -OCH2CO 2 R X , -(CH 2 ) r tetrazole, -(CH 2 ) r oxadiazolone, -(CH 2 ) r tetrazolone, -(CH 2 ) r thiadiazolol, -(CH 2 ) r isoxazol-3-ol, -(CH 2 ) r P(O)(OH)OR x , -(CH 2 ) r S(O) 2 OH, -(CH 2 ) r C(O)NHCN, or -(CH 2 ) r C(O)NHS(O) 2 alkyl;R c is H, C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -OR X , -CO2R X , or NO2;R d is methyl, optionally substituted 5- to 10-membered aryl, optionally substituted 5or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle;each R x is independently at each occurrence hydrogen or C1-C6 alkyl;each R e is independently C1-C6 alkyl, C 2 -C6 alkenyl, C 2 -C6 alkynyl, halogen, -OR y , C1-C6 haloalkyl, -NHR Z , -OH, or -CN;R f is H or absent;each R y and R z is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl;each m and p independently is 0, 1 or 2, wherein m + p 3;q is 0, 1, or 2;r is 0 or 1;and the dotted line is an optional double bond;149 WO 2016/030534 PCT/EP2015/069808 with the proviso that R c is not hydrogen or -CN when X is O, L is -SCH2- and R d is optionally substituted phenyl, R c is not C1-C6 alkyl when X is O, L is -SCH2- and R d is methyl, and that R c is not -CN when X is O, L is -SCH 2 - and R d is 2-furyl.
4
The compound of any one of claims 1-3, wherein:one of R a and R b is hydrogen and the other is CO2R X , CH 2 CO2R X , tetrazole, or oxadiazolone;R c is halogen, -CN, -OR X , or C1-C6 alkyl;R d is methyl, optionally substituted 5- to 10-membered aryl, optionally substituted 5or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle;and R x is hydrogen or C1-C6 alkyl;each R e is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -OR y , C1-C6 haloalkyl, -NHR Z , -OH, or -CN;150 WO 2016/030534 PCT/EP2015/069808 each R y and R z is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl;and n is 0, 1, 2, or 3;with the proviso that R c is not hydrogen or -CN when R d is optionally substituted phenyl and that R c is not -CN when R d is 2-furyl.
5
The compound of any one of claims 1-3, wherein the compound is represented by Formula (II):O R c NH ,co 2 h R d N s (II) or a pharmaceutically acceptable salt thereof.
7
The compound of any one of claims 1-6, wherein R c is -CN or halogen.
8
The compound of any one of claims 1-7, wherein R d is methyl, cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl.
9
The compound of any one of claims 1-7, wherein R d is methyl, cyclohexyl, pyridinyl, thiazolyl, thienyl, or optionally substituted phenyl.
10
The compound of any one of claims 1-4, wherein R a is hydrogen, CH2CO2H, tetrazole, or oxadiazolone (l,2,4-oxadiazol-5(4H)-one).
11
The compound of any one of claims 1-4, wherein R b is hydrogen, CH2CO2H, tetrazole, or oxadiazolone (l,2,4-oxadiazol-5(4H)-one). 151 WO 2016/030534 PCT/EP2015/069808
12
The compound of any one of claims 1-4, wherein n is 0.
16
A pharmaceutical composition comprising a compound of any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, and at least one of a pharmaceutically acceptable carrier, diluent, or excipient.
18
A method of treating a disease or disorder associated with a-amino-βcarboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with ACMSD dysfunction a therapeutically effective amount of one or more compounds of any one of claims 1 to 15.
19
A method of preventing a disease or disorder associated with a-amino-βcarboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with ACMSD dysfunction a therapeutically effective amount of one or more compounds of any one of claims 1 to 15.
20
A method of reducing the risk of a disease or disorder associated with a-amino-βcarboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with ACMSD dysfunction a therapeutically effective amount of one or more compounds of any one of claims 1 to 15.
21
A method of treating a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD + ) levels comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with reduced NAD + levels a therapeutically effective amount of one or more compounds of any one of claims 1 to 15.
22
A method of preventing a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD + ) levels comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with reduced NAD + levels a therapeutically effective amount of one or more compounds of any one of claims 1 to 15.
23
A method of reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD + ) levels comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with reduced 154 WO 2016/030534 PCT/EP2015/069808 NAD + levels a therapeutically effective amount of one or more compounds of any one of claims 1 to 15.
24
A method of treating a disorder associated with mitochondrial dysfunction comprising administering to the subject suffering from or susceptible to developing a metabolic disorder a therapeutically effective amount of one or more compounds of any one of claims 1 to 15 that increases intracellular nicotinamide adenine dinucleotide (NAD + ).
27
A method of promoting oxidative metabolism comprising administering to the subject suffering from or susceptible to developing a metabolic disorder a therapeutically effective amount of one or more compounds of any one of claims 1 to 15 that increases intracellular nicotinamide adenine dinucleotide (NAD + ).
28
A method for the manufacture of a medicament for treating a disease or condition mediated by ACMSD, wherein the medicament comprises a compound of any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof.
29
A pharmaceutical composition for use in a method for treating a disease or condition mediated by ACMSD, wherein the medicament comprises a compound of any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof.
30
Use of a compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating, preventing or reducing the risk of a disease or disorder associated with a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction. 155 WO 2016/030534 PCT/EP2015/069808
31
Use of a compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating, preventing or reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD + ) levels.
32
Use of a compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating, preventing or reducing the risk of a disorder associated with mitochondrial dysfunction.
33
Use of a compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for promoting oxidative metabolism.
34
Use of a compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating, preventing or reducing the risk of a disease or disorder associated with a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction.
35
A compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof for use as a medicament for treating, preventing or reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD + ) levels.
36
A compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof for use as a medicament for treating, preventing or reducing the risk of a disorder associated with mitochondrial dysfunction.
37
A compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof for use as a medicament for promoting oxidative metabolism.
38
A compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof for use in treating, preventing or reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD + ) levels.
39
A compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof for 156 WO 2016/030534 PCT/EP2015/069808 use in for treating, preventing or reducing the risk of a disorder associated with mitochondrial dysfunction.
40
A compound of any of claims 1-15 or a pharmaceutically acceptable salt thereof for use in promoting oxidative metabolism.
41
A method of treating a disease or disorder associated with a-amino-β carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction, comprising administering to a subject in need thereof, a therapeutically effective amount of compound having the one of the following Formula:O O 0 0 157 WO 2016/030534 PCT/EP2015/069808 or a pharmaceutically acceptable salt thereof.
42
Use of a compound having the one of the following Formula:158 WO 2016/030534 PCT/EP2015/069808 159 WO 2016/030534 PCT/EP2015/069808 in the manufacture of a medicament for treating a disease or disorder associated with aamino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction.
43
A compound having the one of the following Formula:160 WO 2016/030534 PCT/EP2015/069808 carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction.
44
A compound having the one of the following Formula:161 WO 2016/030534 PCT/EP2015/069808 162 WO 2016/030534 PCT/EP2015/069808 or a pharmaceutically acceptable salt thereof for use in treating a disease or disorder associated with a-amino-3־carboxymuconate-8semialdehyde decarboxylase (ACMSD) dysfunction.

Independent claims35

1,194 paragraphs in 263 sections, as filed

INHIBITORS OF α-ΑΜΙΝΟ-β-CARBOXYMUCONIC ACID SEMIALDEHYDE DECARBOXYLASE

This application claims the benefit of and priority to U.S. provisional application No. 62/043,853, filed August 29, 2014, the entire contents of which are incorporated herein by reference in its entirety.

FIELD OF THE DISCLOSURE

The present disclosure relates to compounds capable of modulating the activity of aamino־3־carboxymuconic acid semialdehyde decarboxylase (ACMSD). The compounds of the disclosure may be used in methods for the prevention and/or the treatment of diseases and disorders associated with defects in NAD+ biosynthesis, e.g., metabolic disorders, neurodegenerative diseases, chronic inflammatory diseases, kidney diseases, and diseases associated with ageing.

BACKGROUND OF THE DISCLOSURE

ACMSD is a critical enzyme for tryptophan metabolism, and regulates NAD+ biosynthesis from tryptophan. ACMSD is a zinc-dependent amidohydrolase that participates in picolinic acid (PA), quinolinic acid (QA) and NAD homeostasis. ACMSD stands at a branch point of the NAD+ biosynthetic pathway from tryptophan and determines the final fate of the amino acid, i. e., transformation into PA, complete oxidation through the citric acid cycle, or conversion into NAD+ through QA synthesis.

ACMSD has been purified from liver, kidney, and brain human tissues. There are two isoforms ACMSD 1 and ACMSD2 derived from a differential splicing of ACMSD gene transcription but only ACMSD 1 is endowed with enzymatic activity. ACMSD 1 directs ACMS (a-amino-co-carboxymuconic acid semialdehyde) to the acetyl-CoA pathway, and when ACMSD 1 is inhibited, ACMS is non-enzymatically converted to quinolinic acid (QA) leading to the formation of NAD+ and an increase in the intracellular level of NAD+.

Increased levels of NAD+ have been shown to protect against neuronal degeneration, improve muscle function and oxidative metabolism in mice, and enhance lifespan in worms. Whilst reduced levels of NAD+ have been associated with a range of pathophysiological

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PCT/EP2015/069808 states including type 2 diabetes (T2D), hyperlipidemia (elevated cholesterol and TAGs), mitochondrial diseases, neutropenia, cancers, and kidney disorders.

The inhibition of ACMSD thus represents a novel approach to increase NAD+ levels and modify disease pathophysiologies associated with defects in NAD+ biosynthesis.

SUMMARY OF THE DISCLOSURE

It is an object of embodiments of the disclosure to provide novel series of compounds capable of modulating the activity of a-amino-p-carboxymucomc acid semialdehyde decarboxylase (ACMSD), which compounds are useful for the prevention and/or the treatment of diseases and disorders associated with defects in NAD+ biosynthesis, e.g., metabolic disorders, neurodegenerative diseases, chronic inflammatory diseases, kidney diseases, and diseases associated with ageing.

Compounds of Formula (I), as defined herein, may be used in the treatment of a disease or disorder in which ACMSD plays a role. The disclosure features methods of treating a disease or disorder associated with ACMSD dysfunction or with abnormalities in NAD+ biosynthesis by administering to subjects suffering from or susceptible to developing a disease or disorder associated with ACMSD dysfunction a therapeutically effective amount of one or more compounds that increases intracellular NAD+ by ACMSD 1 inhibition, in an amount sufficient to activate sirtuins (SIRTs) and the downstream targets of SIRTs, such as PGC-la, FoxOl and/or superoxide dismutase (SOD). The methods of the present disclosure can be used in the treatment of ACMSD dependent diseases by inhibiting ACMSD. Inhibition of ACMSD may provide a novel approach to the prevention and treatment of metabolic disorders, neurodegenerative diseases, chronic inflammatory diseases, kidney diseases, diseases associated with ageing and other ACMSD dependent diseases, or diseases characterized by defective NAD+ synthesis.

Accordingly, a first aspect of the present disclosure relates to a compound represented by Formula (I):

or a pharmaceutically acceptable salt or tautomer thereof,

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PCT/EP2015/069808 wherein:

X is O, OH, or Cl;

L is -(CH2)mCH2CH2-, -(CH2)mY(CH2)p-, -(CH2)mC(O)(CH2)p-, -(CH2)mC(O)O(CH2)p-, -(CH2)mC(O)NR2(CH2)p-, or -(CH2)mNR2C(O)(CH2)p;

Y is Ο, N or S(O)q;

R1 is C6-C10 aryl or heteroaryl, wherein the aryl and heteroaryl are substituted with Ra and Rb, and optionally substituted with one or more Re;

R2 is H or C1-C6 alkyl;

one of Ra and Rb is hydrogen and the other is -(CH2)rCO2Rx, -OCH2CO2RX,

-(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)rC(O)NHS(O)2alkyl;

Rc is H, C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2RX, or NO2;

Rd is methyl, optionally substituted 5- to 10-membered aryl, optionally substituted 5or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle;

each Rx is independently at each occurrence hydrogen or C1-C6 alkyl;

each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN;

Rf is H or absent;

each Ry and Rz is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl;

each m and p independently is 0, 1 or 2, wherein m + p < 3;

q is 0, 1, or 2;

r is 0 or 1; and the dotted line is an optional double bond;

with the proviso that Rc is not hydrogen or -CN when X is O, L is -SCH2- and Rd is optionally substituted phenyl, Rc is not C1-C6 alkyl when X is O, L is -SCH2- and Rd is methyl, and that Rc is not -CN when X is O, L is -SCH2- and Rd is 2-furyl.

A second aspect of the present disclosure relates to pharmaceutical compositions comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and at least one of a pharmaceutically acceptable carrier, diluent, or excipient.

A third aspect of the disclosure relates to a method of treating a disease or disorder associated with a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction comprising administering to the subject suffering from or susceptible to

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PCT/EP2015/069808 developing a disease or disorder associated with ACMSD dysfunction a therapeutically effective amount of one or more compounds of Formula (I).

A fourth aspect of the disclosure relates to a method of preventing a disease or disorder associated with a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with ACMSD dysfunction a therapeutically effective amount of one or more compounds of Formula (I).

A fifth aspect of the disclosure relates to a method of reducing the risk of a disease or disorder associated with a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with ACMSD dysfunction a therapeutically effective amount of one or more compounds of Formula (1).

A sixth aspect of the disclosure relates to a method of treating a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with reduced NAD+ levels a therapeutically effective amount of one or more compounds of Formula (1).

A seventh aspect of the disclosure relates to a method of preventing a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with reduced NAD+ levels a therapeutically effective amount of one or more compounds of Formula (1).

An eighth aspect of the disclosure relates to a method of reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels comprising administering to the subject suffering from or susceptible to developing a disease or disorder associated with reduced NAD+ levels a therapeutically effective amount of one or more compounds of Formula (I).

An ninth aspect of the disclosure relates to a method of treating a disorder associated with mitochondrial dysfunction comprising administering to the subject suffering from or susceptible to developing a metabolic disorder a therapeutically effective amount of one or more compounds of Formula (I) that increases intracellular nicotinamide adenine dinucleotide (NAD+).

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A tenth aspect of the disclosure relates to a method of promoting oxidative metabolism comprising administering to the subject suffering from or susceptible to developing a metabolic disorder a therapeutically effective amount of one or more compounds of Formula (I) that increases intracellular nicotinamide adenine dinucleotide (NAD+).

An eleventh aspect of the disclosure relates to a method for the manufacture of a medicament for treating a disease or condition mediated by ACMSD, wherein the medicament comprises a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

A twelfth aspect of the disclosure relates to a pharmaceutical composition for use in a method for treating a disease or condition mediated by ACMSD, wherein the medicament comprises a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

A thirteenth aspect of the disclosure relates to a method of treating a disease or disorder associated with a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction, comprising administering to a subject in need thereof, a therapeutically effective amount of compound having one of the following Formulae:

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PCT/EP2015/069808

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PCT/EP2015/069808 or a pharmaceutically acceptable salt thereof.

A fourteenth aspect of the disclosure relates to the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating, preventing or reducing the risk of a disease or disorder associated with a-amino-βcarboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction.

A fifteenth aspect of the disclosure relates to the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating, preventing or reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

A sixteenth aspect of the disclosure relates to the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating, preventing or reducing the risk of a disorder associated with mitochondrial dysfunction.

A seventeenth aspect of the disclosure relates to the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for promoting oxidative metabolism.

A eighteenth aspect of the disclosure relates to the use of a compound of Formula (I), a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating, preventing or reducing the risk of a disease or disorder associated with a-amino-βcarboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction.

A nineteenth aspect of the disclosure relates to a compound of Formula (I), or a pharmaceutically acceptable salt thereof for use as a medicament for treating, preventing or reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

A twentieth aspect of the disclosure relates to a compound of Formula (I), or a pharmaceutically acceptable salt thereof for use as a medicament for treating, preventing or reducing the risk of a disorder associated with mitochondrial dysfunction.

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PCT/EP2015/069808 for use as a medicament for treating a disease or disorder associated with a-amino-βcarboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction.

A twenty sixth aspect of the disclosure relates to a compound having the one of the

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or a pharmaceutically acceptable salt thereof for use in treating a disease or disorder associated with a-amino-3־carboxymuconate-8semialdehyde decarboxylase (ACMSD) dysfunction.

In certain aspects, the ACMSD modulating compounds may be administered alone or in combination with other compounds, including other ACMSD modulating compounds, or other therapeutic agents.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the specification, the singular forms also include the plural unless the context clearly dictates otherwise. Although methods and materials similar to or equivalent to those described herein can be used in the practice and testing of the disclosure, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference. The references cited herein are not admitted to be prior art to the claimed disclosure. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

Other features and advantages of the disclosure will be apparent from the following

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PCT/EP2015/069808 detailed description and claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph of the measured NAD+ levels in human primary hepatocytes treated with Compound 4, detected by LC-MS/MS.

FIG. 2 is a graph of the measured NAD+ levels in murine primary hepatocytes treated with different concentrations of Compound 17 for 24 hours and detected by LC-MS/MS. The data indicate an increase in NAD+ levels in murine primary hepatocytes treated with Compound 17.

FIG. 3 is a graph of the measured NAD+ content in human primary hepatocytes treated with various concentrations of Compound 1 and mono-(2-ethylhexyl)phthalate (MEHP), as a control.

FIG. 4A is a graph of gene expression of Acmsd, Sod-1, and Sod-2 as determined by RT-qPCR in AML-12 cells treated with Compound 1 for 24 hours. FIG. 4B is a graph of gene expression of Sod-1 and Sod-2 as determined by RT-qPCR in Hepa-1.6 cells treated with Compound 1 for 24 hours. FIG. 4C is a graph of gene expression of Acmsd, Sod-1, Sod2, and Pgcla as determined by RT-qPCR in primary mouse hepatocytes treated with Compound 17 for 24 hours. Bar graphs represent mean ± SEM, ***p < 0.005

FIG. 5A is a graph showing the modulation of SOD2 activity in AML-12 cells treated for 24 hours with Compound 1. FIG. 5B shows a graph of the modulation of SOD2 activity in AML-12 cells treated for 24 hours with Compound 17. FIG. 5C shows a graph of the modulation of SOD2 activity in primary murine hepatocytes cells treated for 24 hours with Compound 17.

FIG. 6A is a gel showing the effect of Compound 1 on the FoxOl phosphorylation levels. FIG. 6B is a gel showing the effect of Compound 17 on the FoxOl phosphorylation levels.

FIG. 7A is a graph of changes in acsmd-1 and sod-3 expression at mRNA levels measured in N2 wild type worms at day 2 of adulthood by acmsd-1 RNAi silencing in Caenorhabditis elegans (C. elegans). FIG. 7B is a graph of the induction of sod-3 expression at protein levels in N2 worms at day 3 of adulthood, quantified by using SOD-3 gfp reporter strain, after acmsd-1 RNAi silencing in C. elegans. FIG. 7C is a graph showing the survival of worms upon downregulation of acmsd-1 by feeding a specific RNAi in C. elegans. FIG.

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7C shows that downregulation of acmsd-1 improves the survival of worms in a SIR-2.1 and DAF-16 dependent manner. FIG. 7D is a graph showing that downregulation of acmsd-1 improves the stress-resistance of worms when they are exposed to paraquat-induced oxidative stress. FIG. 7E is a graph showing the mobility of worms over time fed with acmsd-1 RNAi under paraquat-induced oxidative stress condition. As FIGs. 7C-7E show, reduced acmsd-1 expression improves the survival and fitness of worms under paraquat-induced oxidative stress. FIG. 7F is a graph that shows the survival of worms under paraquat-induced stress conditions when exposed to acmsd-1 RNAi during different stages of development. FIG. 7F illustrates that the improvement of the survival of worms under paraquat conditions is independent of the developmental stage at which the worms were exposed to the acmsd-1 RNAi. FIG. 7G is a graph showing worm survival under paraquat-induced stress conditions upon downregulation of acmsd-1 combined with daf-16 downregulation by feeding a specific RNAi in C. elegans. FIG. 7G shows that the improved survival of worms with downregulated acmsd-1 is daf-16 dependent under paraquat-induced oxidative stress conditions.

FIG. 8A is a graph of changes in caspase3/7 activity induced by cisplatin in MDCK cells when treated with different concentrations of Compound 18 in combination with cisplatin. FIG. 8B is a graph of changes in caspase3/7 activity induced by cisplatin in MDCK cells when treated with different concentrations of Compound 18 one hour prior to the addition of cisplatin.

DETAILED DESCRIPTION OF THE DISCLOSURE

Compounds of Formula (I)

The present disclosure relates to compounds of Formula (I):

X R° JL .Rf 'N ^R1

Rd N r (I) or a pharmaceutically acceptable salt or tautomer thereof, wherein:

X is O, OH, or Cl;

L is -(CH2)mCH2CH2-, -(CH2)mY(CH2)p-, -(CH2)mC(O)(CH2)p-,

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-(CH2)mC(O)O(CH2)p-, -(CH2)mC(O)NR2(CH2)p-, or -(CH2)mNR2C(O)(CH2)p;

Y is Ο, N or S(O)q;

R1 is C6-C10 aryl or heteroaryl, wherein the aryl and heteroaryl are substituted with Ra and Rb, and optionally substituted with one or more Re;

R2 is H or C1-C6 alkyl;

one of Ra and Rb is hydrogen and the other is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)rC(O)NHS(O)2alkyl;

Rc is H, C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2RX, or NO2;

Rd is methyl, optionally substituted 5- to 10-membered aryl, optionally substituted 5or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle;

each Rx is independently at each occurrence hydrogen or C1-C6 alkyl;

each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN;

R is H or absent;

each Ry and Rz is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl;

each m and p independently is 0, 1 or 2, wherein m + p < 3;

q is 0, 1, or 2;

r is 0 or 1; and the dotted line is an optional double bond;

with the proviso that Rc is not hydrogen or -CN when X is O, L is -SCH2- and Rd is optionally substituted phenyl, Rc is not C1-C6 alkyl when X is O, L is -SCH2- and Rd is methyl, and that Rc is not -CN when L is -SCH2- and Rd is 2-furyl.

In some embodiments of Formula (I), X is O, OH, or Cl. In other embodiments, X is O. In other embodiments, X is OH. In other embodiments, X is Cl.

In some embodiments of Formula (I), L is -(CH2)mCH2CH2-, -(CH2)mY(CH2)p-, -(CH2)mC(O)(CH2)p-, -(CH2)mC(O)O(CH2)p-, -(CH2)mC(O)NR2(CH2)p-, or-(CH2)mNR2C(O) (CH2)p. In other embodiments, L is -CH2CH2-, -CH2CH2CH2-,-SCH2-, -SCH2CH2-, -CH2S-, -CH2SCH2-, -CH2CH2S-, -S(O)CH2-, -S(O)CH2CH2-, -CH2S(O)-, -CH2S(O)CH2-, -CH2CH2S(O)-, -S(O)2CH2-, -S(O)2CH2CH2-, -CH2S(O)2-, -CH2S(O)2CH2-, -CH2CH2S(O)2-, -OCH2-, -OCH2CH2-, -CH2O-, -CH2OCH2-, -CH2CH2O-,-NR2CH2-, - CH2NR2-, -CH2NR2CH2-, - CH2CH2NR2-, -NR2CH2CH2-, -C(O)CH2-, -C(O)CH2CH2-,

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-C(O)O-, -C(O)OCH2-, -CH2C(O)O-, -C(O)NR2-, -C(O)NR2CH2-, -NR2C(O), -NR2C(O)CH2, or -CH2NR2C(O). In other embodiments, L is -CH2CH2-, -CH2CH2CH2-, -SCH2-, -SCH2CH2-, -S(O)CH2-, -S(O)CH2CH2-,-S(O)2CH2-, -S(O)2CH2CH2-, -OCH2-, -OCH2CH2-, -NR2CH2-, -NR2CH2CH2-, -C(O)CH2-, -C(O)CH2CH2-, -C(O)O-, -C(O)OCH2,-CH2C(O)O-, -C(O)NR2-, -C(O)NR2CH2-, - NR2C(O), or - NR2C(O)CH2. In other embodiments, L is -CH2CH2-, -CH2C(O)-, -C(O)CH2-, -NR2CH2-, -CH2NR2-, -OCH2-, -CH2O-, -SCH2-, -CH2S-, -S(O)CH2-, -CH2S(O)-, -CH2S(O)2-, or -S(O)2CH2-.

In some embodiments of Formula (I), R1 is C6־C!0 aryl or heteroaryl, wherein the aryl and heteroaryl are substituted with Ra and Rb, and optionally substituted with one or more Re. In other embodiments, R1 is C6־C!0 aryl substituted with Ra and Rb, and optionally substituted with one or more Re. In other embodiments, R1 is heteroaryl substituted with Ra and Rb, and optionally substituted with one or more Re. In further embodiments, R1 is phenyl substituted with Ra and Rb, and optionally substituted with one or more Re.

In some embodiments of Formula (I), Ra is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)rC(O)NHS(O)2alkyl. In other embodiments, Ra is -(CH2)rCO2Rx, -OCH2CO2Rx, tetrazole, -(CH2)tetrazole, oxadiazolone, -(CH2)oxadiazolone, tetrazolone, -(CH2)tetrazolone, thiadiazolol, -(CH2)thiadiazolol, isoxazol-3-ol, -(CH2) isoxazol-3-ol, -P(O)(OH)ORX, -(CH2)P(O)(OH)ORx, -S(O)2OH , -(CH2)S(O)2OH, -C(O)NHCN -(CH2)C(O)NHCN, -C(O)NHS(O)2alkyl, or -(CH2)C(O)NHS(O)2alkyl. In other embodiments, Ra is hydrogen, CO2RX, CH2CO2RX, tetrazole, or oxadiazolone. In further embodiments, Ra is hydrogen, CO2H, CH2CO2H, tetrazole, or l,2,4-oxadiazol-5(4H)-one.

In some embodiments of Formula (I), Rb is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)rC(O)NHS(O)2alkyl. In other embodiments, Rb is -(CH2)rCO2Rx, -OCH2CO2Rx, tetrazole, -(CH2)tetrazole, oxadiazolone, -(CH2)oxadiazolone, tetrazolone, -(CH2)tetrazolone, thiadiazolol, -(CH2)thiadiazolol, isoxazol-3-ol, -(CH2) isoxazol-3-ol, -P(O)(OH)ORX, -(CH2)P(O)(OH)ORx, -S(O)2OH , -(CH2)S(O)2OH, -C(O)NHCN -(CH2)C(O)NHCN, -C(O)NHS(O)2alkyl, or -(CH2)C(O)NHS(O)2alkyl. In other embodiments, Rb is hydrogen, CO2RX, CH2CO2RX, tetrazole, or oxadiazolone. In further

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PCT/EP2015/069808 embodiments, Rb is hydrogen, CO2H, CH2CO2H, tetrazole, or l,2,4-oxadiazol-5(4H)-one. In further embodiments, Rb is hydrogen.

In some embodiments of Formula (I), Rc is H, C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2Rx, or NO2. In other embodiments, Rc is C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2RX, orNO2. In other embodiments, Rcis halogen, -CN, -ORX, or C1-C6 alkyl. In other embodiments, Rc is halogen, -CN, -ORX, or C1-C3 alkyl. In other embodiments, Rc is H, -CN, or halogen. In other embodiments, Rc is -CN or halogen.

In some embodiments of Formula (I), Rd is methyl, optionally substituted 5- to 10membered aryl, optionally substituted 5- or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle. In other embodiments, Rd is methyl, optionally cyclohexyl, optionally substituted pyridinyl, optionally substituted thiazolyl, optionally substituted phenyl, or optionally substituted thienyl. In other embodiments, Rd is methyl, cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more substituents independently selected from halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, -OH, CN, and amino. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more substituents independently selected from halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more halogen. In yet other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, 4-chlorophenyl, 4-methylphenyl, or thienyl.

In some embodiments of Formula (I), each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN. In other embodiments, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, halogen, -ORy, C1-C4 haloalkyl, NHRZ,-OH, or-CN.

f f

In some embodiments of Formula (I), R is H or absent. In other embodiments, R is £

H. In other embodiments, R is absent, when N to which it is attached participates in a double bond.

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In some embodiments of Formula (I), Rx is hydrogen or C1-C6 alkyl. In other embodiments, Rx is hydrogen or C1-C3 alkyl. In further embodiments, Rx is hydrogen, methyl, ethyl, n-propyl, or iso-propyl.

In some embodiments of Formula (I), Ry is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl. In other embodiments, Ry is hydrogen, C1-C3 alkyl, or C1-C3 haloalkyl.

In some embodiments of Formula (I), each Rz is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl. In other embodiments, Rz is hydrogen, C1-C3 alkyl, or C1-C3 haloalkyl.

In some embodiments of Formula (I), m is 0, 1 or 2. In other embodiments, m is 0.

In other embodiments, m is 1. In yet other embodiments, m is 2.

In some embodiments of Formula (I), p is 0, 1 or 2. In other embodiments, p is 0. In other embodiments, p is 1. In yet other embodiments, p is 2.

In some embodiments of Formula (I), m + p < 3;

In some embodiments of Formula (I), q is 0, 1, or 2. In other embodiments, q is 0. In other embodiments, q is 1. In other embodiments, q is 2.

In some embodiments of Formula (I), r is 0 or 1. In other embodiments, r is 0. In other embodiments, r is 1.

In some embodiments of Formula (I), the dotted line is a single bond. In other embodiments, the dotted line is a double bond.

In some embodiments of Formula (I), one of Ra and Rb is hydrogen and the other is CO2Rx, CH2CO2Rx, tetrazole, or oxadiazolone. In other embodiments, Rb is hydrogen and Ra is CH2CO2H, tetrazole, or (l,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (I), Rb is hydrogen, Rc is -CN, Rd is thienyl, and Ra is CH2CO2H, tetrazole, or (l,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (I), Rc is halogen, Ra is -CO2H, and Rb is H. In other embodiments, Rc is -Br, Ra is -CO2H, and Rb is H. In further embodiments, Rc is -Cl, Ra is -CO2H, and Rb is H.

In some embodiments of Formula (I), Rc is halogen, Ra is tetrazole, and Rb is H. In other embodiments, Rc is -Br, Ra is tetrazole, and Rb is H. In further embodiments, Rc is -Cl, Ra is tetrazole, and Rb is H.

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In some embodiments of Formula (I), Rc is halogen, Ra is -CH2CO2H, and Rb is H. In other embodiments, Rc is -Br, Ra is -CH2CO2H, and Rb is H. In further embodiments, Rc is Cl, Ra is -CH2CO2H, and Rb is H.

In some embodiments of Formula (I), Rc is halogen, Ra is (l,2,4-oxadiazol-5(4H)one), and Rb is H. In other embodiments, Rc is -Br, Ra is (l,2,4-oxadiazol-5(4H)-one), and Rb is H. In other embodiments, Rc is -Cl, Ra is (l,2,4-oxadiazol-5(4H)-one), and Rb is H.

In some embodiments of Formula (I), Rc is -CN, Ra is -CO2H, and Rb is H. In other embodiments, Rc is -CN, Ra is -CH2CO2H, and Rb is H. In other embodiments, Rc is -CN, Ra is tetrazole, and Rb is H. In yet other embodiments, Rc is -CN, Ra is (l,2,4-oxadiazol-5(4H)one), and Rb is H.

In some embodiments of Formula (I), Rc is not hydrogen or -CN and X is O, L is SCH2- and Rd is optionally substituted phenyl. In other embodiments, Rc is not C1-C6 alkyl and X is O, L is -SCH2- and Rd is methyl. In other embodiments, Rc is not -CN and X is O, L is -SCH2- and Rd is 2-furyl.

In some embodiments of Formula (I), Rc is not hydrogen or -CN when X is O, L is SCH2- and Rd is optionally substituted phenyl.

In some embodiments of Formula (I), Rc is not C1-C6 alkyl when X is O, L is -SCH2and Rd is methyl.

In some embodiments of Formula (I), Rc is not -CN when X is 0, L is -SCH2- and Rd is 2-furyl.

In one embodiment, the compound of Formula (I) is represented by Formula (la):

or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

L is -(CH2)mCH2CH2-, -(CH2)mY(CH2)p-, -(CH2)mC(O)(CH2)p-, -(CH2)mC(O)O(CH2)p-, -(CH2)mC(O)NR2(CH2)p-, or -(CH2)mNR2C(O)(CH2)p;

Y is Ο, N or S(0)q;

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R1 is C6-C10 aryl or heteroaryl, wherein the aryl and heteroaryl are substituted with Ra and Rb, and optionally substituted with one or more Re;

R2 is H or C1-C6 alkyl;

Rc is H, C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2RX, or NO2;

Rd is methyl, optionally substituted 5- to 10-membered aryl, optionally substituted 5or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle;

each Rx is independently at each occurrence hydrogen or C1-C6 alkyl;

each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN;

each Ry and Rz is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl;

each m and p independently is 0, 1 or 2, wherein m + p < 3;

q is 0, 1, or 2; and r is 0 or 1;

with the proviso that Rc is not hydrogen or -CN when L is -SCH2- and Rd is optionally substituted phenyl, Rc is not C1-C6 alkyl when L is -SCH2- and Rd is methyl, and that Rc is not -CN when L is -SCH2- and Rd is 2-furyl.

In some embodiments of Formula (la),

L is -CH2CH2-, -CH2C(O)-, -C(O)CH2-, -NR2CH2-, -CH2NR2-, -OCH2-, -CH2O-, -SCH2-, -CH2S-, -S(O)CH2-, -CH2S(O)-, -CH2S(O)2-, or -S(O)2CH2-;

Y is Ο, N or S(O)q;

R1 is C6-C10 aryl or heteroaryl, wherein the aryl and heteroaryl are substituted with Ra and Rb, and optionally substituted with one or more Re;

R2 is H or C1-C6 alkyl;

Rc is C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2RX, or NO2;

Rd is methyl, optionally substituted 5- to 10-membered aryl, optionally substituted 5or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle;

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PCT/EP2015/069808 each Rx is independently at each occurrence hydrogen or C1-C6 alkyl;

each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN;

each Ry and Rz is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl;

each m and p independently is 0, 1 or 2, wherein m + p < 3;

q is 0, 1, or 2; and r is 0 or 1;

with the proviso that Rc is not -CN when L is -SCH2- and Rd is optionally substituted phenyl, Rc is not C1-C6 alkyl when L is -SCH2- and Rd is methyl, and that Rc is not -CN when L is -SCH2- and Rd is 2-furyl.

In some embodiments of Formula (la), L is -(CH2)mCH2CH2-, -(CH2)mY(CH2)p-, -(CH2)mC(O)(CH2)p-, -(CH2)mC(O)O(CH2)p-, -(CH2)mC(O)NR2(CH2)p-, or-(CH2)mNR2C(O) (CH2)p. In other embodiments, L is -CH2CH2-, -CH2CH2CH2-,-SCH2-, -SCH2CH2-, -CH2S-, -CH2SCH2-, -CH2CH2S-, -S(O)CH2-, -S(O)CH2CH2-, -CH2S(O)-, -CH2S(O)CH2-, -CH2CH2S(O)-, -S(O)2CH2-, -S(O)2CH2CH2-, -CH2S(O)2-, -CH2S(O)2CH2-, -CH2CH2S(O)2-, -OCH2-, -OCH2CH2-, -CH2O-, -CH2OCH2-, -CH2CH2O-,-NR2CH2-, - CH2NR2-, -CH2NR2CH2-, - CH2CH2NR2-, -NR2CH2CH2-, -C(O)CH2-, -C(O)CH2CH2-, -C(O)O-, -C(O)OCH2-, -CH2C(O)O-, -C(O)NR2-, -C(O)NR2CH2-, -NR2C(O), -NR2C(O)CH2, or -CH2NR2C(O). In other embodiments, L is -CH2CH2-, -CH2CH2CH2-, -SCH2-, -SCH2CH2-, -S(O)CH2-, -S(O)CH2CH2-,-S(O)2CH2-, -S(O)2CH2CH2-, -OCH2-, -OCH2CH2-, -NR2CH2-, -NR2CH2CH2-, -C(O)CH2-, -C(O)CH2CH2-, -C(O)O-, -C(O)OCH2,-CH2C(O)O-, -C(O)NR2-, -C(O)NR2CH2-, - NR2C(O), or - NR2C(O)CH2. In other embodiments, L is -CH2CH2-, -CH2C(O)-, -C(O)CH2-, -NR2CH2-, -CH2NR2-, -OCH2-, -CH2O-, -SCH2-, -CH2S-, -S(O)CH2-, -CH2S(O)-, -CH2S(O)2-, or -S(O)2CH2-.

In some embodiments of Formula (la), R1 is C6־C!0 aryl or heteroaryl, wherein the aryl and heteroaryl are substituted with Ra and Rb, and optionally substituted with one or more Re. In other embodiments, R1 is C6-C10 aryl substituted with Ra and Rb, and optionally substituted with one or more Re. In other embodiments, R1 is heteroaryl substituted with Ra and Rb, and optionally substituted with one or more Re. In further embodiments, R1 is phenyl substituted with Ra and Rb, and optionally substituted with one or more Re.

In some embodiments of Formula (la), Ra is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or

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-(CH2)rC(O)NHS(O)2alkyl. In other embodiments, Ra is -(CH2)rCO2Rx, -OCH2CO2Rx, tetrazole, -(CH2)tetrazole, oxadiazolone, -(CH2)oxadiazolone, tetrazolone, -(CH2)tetrazolone, thiadiazolol, -(CH2)thiadiazolol, isoxazol-3-ol, -(CH2) isoxazol-3-ol, -P(O)(OH)ORX, -(CH2)P(O)(OH)ORx, -S(O)2OH , -(CH2)S(O)2OH, -C(O)NHCN -(CH2)C(O)NHCN, -C(O)NHS(O)2alkyl, or -(CH2)C(O)NHS(O)2alkyl. In other embodiments, Ra is hydrogen, CO2RX, CH2CO2RX, tetrazole, or oxadiazolone. In further embodiments, Ra is hydrogen, CO2H, CH2CO2H, tetrazole, or l,2,4-oxadiazol-5(4H)-one.

In some embodiments of Formula (la), Rb is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)rC(O)NHS(O)2alkyl. In other embodiments, Rb is -(CH2)rCO2Rx, -OCH2CO2Rx, tetrazole, -(CH2)tetrazole, oxadiazolone, -(CH2)oxadiazolone, tetrazolone, -(CH2)tetrazolone, thiadiazolol, -(CH2)thiadiazolol, isoxazol-3-ol, -(CH2) isoxazol-3-ol, -P(O)(OH)ORX, -(CH2)P(O)(OH)ORx, -S(O)2OH , -(CH2)S(O)2OH, -C(O)NHCN -(CH2)C(O)NHCN, -C(O)NHS(O)2alkyl, or -(CH2)C(O)NHS(O)2alkyl. In other embodiments, Rb is hydrogen, CO2RX, CH2CO2RX, tetrazole, or oxadiazolone. In further embodiments, Rb is hydrogen, CO2H, CH2CO2H, tetrazole, or l,2,4-oxadiazol-5(4H)-one. In further embodiments, Rb is hydrogen.

In some embodiments of Formula (la), Rc is H, C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2Rx, or NO2. In other embodiments, Rc is C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2RX, orNO2. In other embodiments, Rcis halogen, -CN, -ORX, or C1-C6 alkyl. In other embodiments, Rc is halogen, -CN, -ORX, or C1-C3 alkyl. In other embodiments, Rc is H, -CN, or halogen. In other embodiments, Rc is -CN or halogen.

In some embodiments of Formula (la), Rd is methyl, optionally substituted 5- to 10membered aryl, optionally substituted 5- or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle. In other embodiments, Rd is methyl, optionally cyclohexyl, optionally substituted pyridinyl, optionally substituted thiazolyl, optionally substituted phenyl, or optionally substituted thienyl. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more substituents independently selected from halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, -OH, CN, and amino. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more substituents independently selected from halogen, C!-C6 alkyl, C!-C6

PCT/EP2015/069808 each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN;

with the proviso that Rc is not hydrogen or -CN when Rd is optionally substituted phenyl, Rc is not C1-C6 alkyl when Rd is methyl, and that Rc is not -CN when Rd is 2-furyl.

In some embodiments of formula (lb), one of Ra and Rb is hydrogen and the other is CO2RX, CH2CO2RX, tetrazole, or oxadiazolone;

Rc is halogen, -CN, -ORX, or C1-C6 alkyl;

Rd is methyl, optionally substituted 5- to 10-membered aryl, optionally substituted 5or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle; and

Rx is hydrogen or C1-C6 alkyl;

each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN;

each Ry and Rz is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl; and n is 0, 1, 2, or 3;

with the proviso that Rc is not -CN when Rd is optionally substituted phenyl, Rc is not C1-C6 alkyl when Rd is methyl, and that Rc is not -CN when Rd is 2-furyl.

In some embodiments of Formula (lb), Ra is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)rC(O)NHS(O)2alkyl. In other embodiments, Ra is -(CH2)rCO2Rx, -OCH2CO2Rx, tetrazole, -(CH2)tetrazole, oxadiazolone, -(CH2)oxadiazolone, tetrazolone, -(CH2)tetrazolone, thiadiazolol, -(CH2)thiadiazolol, isoxazol-3-ol, -(CH2) isoxazol-3-ol, -P(O)(OH)ORX, -(CH2)P(O)(OH)ORx, -S(O)2OH , -(CH2)S(O)2OH, -C(O)NHCN -(CH2)C(O)NHCN, -C(O)NHS(O)2alkyl, or -(CH2)C(O)NHS(O)2alkyl. In other embodiments, Ra is hydrogen, CO2RX, CH2CO2RX, tetrazole, or oxadiazolone. In further embodiments, Ra is hydrogen, CO2H, CH2CO2H, tetrazole, or l,2,4-oxadiazol-5(4H)-one.

In some embodiments of Formula (lb), Rb is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)rC(O)NHS(O)2alkyl. In other embodiments, Rb is -(CH2)rCO2Rx, -OCH2CO2Rx, tetrazole, -(CH2)tetrazole, oxadiazolone, -(CH2)oxadiazolone, tetrazolone, -(CH2)tetrazolone, thiadiazolol, -(CH2)thiadiazolol, isoxazol-3-ol, -(CH2) isoxazol-3-ol,

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-P(O)(OH)ORX, -(CH2)P(O)(OH)ORx, -S(O)2OH , -(CH2)S(O)2OH, -C(O)NHCN -(CH2)C(O)NHCN, -C(O)NHS(O)2alkyl, or -(CH2)C(O)NHS(O)2alkyl. In other embodiments, Rb is hydrogen, CO2RX, CH2CO2RX, tetrazole, or oxadiazolone. In further embodiments, Rb is hydrogen, CO2H, CH2CO2H, tetrazole, or l,2,4-oxadiazol-5(4H)-one. In further embodiments, Rb is hydrogen.

In some embodiments of Formula (lb), Rcis H, C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2Rx, or NO2. In other embodiments, Rc is C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2RX, orNO2. In other embodiments, Rcis halogen, -CN, -ORX, or C1-C6 alkyl. In other embodiments, Rc is halogen, -CN, -ORX, or C1-C3 alkyl. In other embodiments, Rc is H, -CN, or halogen. In other embodiments, Rc is -CN or halogen.

In some embodiments of Formula (lb), Rd is methyl, optionally substituted 5- to 10membered aryl, optionally substituted 5- or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle. In other embodiments, Rd is methyl, optionally cyclohexyl, optionally substituted pyridinyl, optionally substituted thiazolyl, optionally substituted phenyl, or optionally substituted thienyl. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more substituents independently selected from halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, -OH, CN, and amino. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more substituents independently selected from halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more halogen. In other embodiments, Rd is methyl, cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl. In yet other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, 4-chlorophenyl, 4-methylphenyl, or thienyl.

In some embodiments of Formula (lb), each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN. In other embodiments, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, halogen, -ORy, C1-C4 haloalkyl, NHRZ,-OH, or-CN.

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In some embodiments of Formula (lb), Rx is hydrogen or C1-C6 alkyl. In other embodiments, Rx is hydrogen or C1-C3 alkyl. In further embodiments, Rx is hydrogen, methyl, ethyl, n-propyl, or iso-propyl.

In some embodiments of Formula (lb), Ry is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl. In other embodiments, Ry is hydrogen, C1-C3 alkyl, or C1-C3 haloalkyl.

In some embodiments of Formula (lb), each Rz is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl. In other embodiments, Rz is hydrogen, C1-C3 alkyl, or C1-C3 haloalkyl.

In some embodiments of Formula (lb), n is 0, 1, 2, or 3. In other embodiments, n is 0 or 1. In further embodiments, n is 0.

In some embodiments of Formula (lb), one of Ra and Rb is hydrogen and the other is CO2Rx, CH2CO2Rx, tetrazole, or oxadiazolone. In other embodiments, Rb is hydrogen and Ra is CH2CO2H, tetrazole, or (l,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (lb), Rbis hydrogen, Rcis -CN, Rdis thienyl, and Ra is CH2CO2H, tetrazole, or (l,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (lb), Rc is halogen, Ra is -CO2H, and Rb is H. In other embodiments, Rc is -Br, Ra is -CO2H, and Rb is H. In further embodiments, Rc is -Cl, Ra is -CO2H, and Rb is H.

In some embodiments of Formula (lb), Rc is halogen, Ra is tetrazole, and Rb is H. In other embodiments, Rc is -Br, Ra is tetrazole, and Rb is H. In further embodiments, Rc is -Cl, Ra is tetrazole, and Rb is H.

In some embodiments of Formula (lb), Rc is halogen, Ra is -CH2CO2H, and Rb is H. In other embodiments, Rc is -Br, Ra is -CH2CO2H, and Rb is H. In further embodiments, Rc is -Cl, Ra is -CH2CO2H, and Rb is H.

In some embodiments of Formula (lb), Rc is halogen, Ra is (l,2,4-oxadiazol-5(4H)one), and Rb is H. In other embodiments, Rc is -Br, Ra is (l,2,4-oxadiazol-5(4H)-one), and Rb is H. In other embodiments, Rc is -Cl, Ra is (l,2,4-oxadiazol-5(4H)-one), and Rb is H.

In some embodiments of Formula (lb), Rc is -CN, Ra is -CO2H, and Rb is H. In other embodiments, Rc is -CN, Ra is -CH2CO2H, and Rb is H. In other embodiments, Rc is -CN, Ra is tetrazole, and Rb is H. In yet other embodiments, Rc is -CN, Ra is (l,2,4-oxadiazol-5(4H)one), and Rb is H.

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In some embodiments of Formula (lb), Rc is not hydrogen or -CN and Rd is optionally substituted phenyl. In other embodiments, Rc is not C1-C6 alkyl and Rd is methyl. In other embodiments, Rc is not -CN and Rd is 2-furyl.

In some embodiments of Formula (lb), Rc is not hydrogen or -CN when and Rd is optionally substituted phenyl.

PCT/EP2015/069808 n is 0, 1, 2, or 3.

In some embodiments of Formula (III), one of Ra and Rb is hydrogen and the other is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)r C(O)NHS(O)2alkyl;

Rc is C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2RX, or NO2;

Rd is methyl, optionally substituted 5- to 10-membered aryl, optionally substituted 5or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle;

each Rx is independently at each occurrence hydrogen or C1-C6 alkyl;

each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN.

In some embodiments of formula (III), one of Ra and Rb is hydrogen and the other is CO2RX, CH2CO2RX, tetrazole, or oxadiazolone;

Rc is halogen, -CN, -ORX, or C1-C6 alkyl;

Rd is methyl, optionally substituted 5- to 10-membered aryl, optionally substituted 5or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle; and

Rx is hydrogen or C1-C6 alkyl;

each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN;

each Ry and Rz is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl; and n is 0, 1, 2, or 3;

with the proviso that Rc is not hydrogen or -CN when Rd is optionally substituted phenyl and that Rc is not -CN when Rd is 2-furyl.

In some embodiments of Formula (III), Ra is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)rC(O)NHS(O)2alkyl. In other embodiments, Ra is -(CH2)rCO2Rx, -OCH2CO2Rx, tetrazole, -(CH2)tetrazole, oxadiazolone, -(CH2)oxadiazolone, tetrazolone, -(CH2)tetrazolone, thiadiazolol, -(CH2)thiadiazolol, isoxazol-3-ol, -(CH2) isoxazol-3-ol, -P(O)(OH)ORX, -(CH2)P(O)(OH)ORx, -S(O)2OH , -(CH2)S(O)2OH, -C(O)NHCN

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-(CH2)C(O)NHCN, -C(O)NHS(O)2alkyl, or -(CH2)C(O)NHS(O)2alkyl. In other embodiments, Ra is hydrogen, CO2RX, CH2CO2RX, tetrazole, or oxadiazolone. In further embodiments, Ra is hydrogen, CO2H, CH2CO2H, tetrazole, or l,2,4-oxadiazol-5(4H)-one.

In some embodiments of Formula (III), Rb is -(CH2)rCO2Rx, -OCH2CO2RX, -(CH2)rtetrazole, -(CH2)roxadiazolone, -(CH2)rtetrazolone, -(CH2)rthiadiazolol, -(CH2)r isoxazol-3-ol, -(CH2)rP(O)(OH)ORx, -(CH2)rS(O)2OH, -(CH2)rC(O)NHCN, or -(CH2)rC(O)NHS(O)2alkyl. In other embodiments, Rb is -(CH2)rCO2Rx, -OCH2CO2Rx, tetrazole, -(CH2)tetrazole, oxadiazolone, -(CH2)oxadiazolone, tetrazolone, -(CH2)tetrazolone, thiadiazolol, -(CH2)thiadiazolol, isoxazol-3-ol, -(CH2) isoxazol-3-ol, -P(O)(OH)ORX, -(CH2)P(O)(OH)ORx, -S(O)2OH , -(CH2)S(O)2OH, -C(O)NHCN -(CH2)C(O)NHCN, -C(O)NHS(O)2alkyl, or -(CH2)C(O)NHS(O)2alkyl. In other embodiments, Rb is hydrogen, CO2RX, CH2CO2RX, tetrazole, or oxadiazolone. In further embodiments, Rb is hydrogen, CO2H, CH2CO2H, tetrazole, or l,2,4-oxadiazol-5(4H)-one. In further embodiments, Rb is hydrogen.

In some embodiments of Formula (III), Rcis H, C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2Rx, or NO2. In other embodiments, Rc is C1-C6 alkyl, C1-C6 haloalkyl, halogen, -CN, -ORX, -CO2RX, orNO2. In other embodiments, Rcis halogen, -CN, -ORX, or C1-C6 alkyl. In other embodiments, Rc is halogen, -CN, -ORX, or C1-C3 alkyl. In other embodiments, Rc is H, -CN, or halogen. In other embodiments, Rc is -CN or halogen.

In some embodiments of Formula (III), Rd is methyl, optionally substituted 5- to 10membered aryl, optionally substituted 5- or 6-membered heteroaryl, or optionally substituted 5- or 6-membered carbocycle. In other embodiments, Rd is methyl, optionally cyclohexyl, optionally substituted pyridinyl, optionally substituted thiazolyl, optionally substituted phenyl, or optionally substituted thienyl. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more substituents independently selected from halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, -OH, CN, and amino. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more substituents independently selected from halogen, C1-C6 alkyl, C1-C6 hydroxyalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy. In other embodiments, Rd is cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each is optionally substituted with one or more halogen. In other embodiments, Rd is methyl, cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl. In yet other embodiments, Rd is cyclohexyl,

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In some embodiments of Formula (III), each Re is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, -ORy, C1-C6 haloalkyl, -NHRZ, -OH, or -CN. In other embodiments, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, halogen, -ORy, C1-C4 haloalkyl, NHRZ,-OH, or-CN.

In some embodiments of Formula (III), Rx is hydrogen or C1-C6 alkyl. In other embodiments, Rx is hydrogen or C1-C3 alkyl. In further embodiments, Rx is hydrogen, methyl, ethyl, n-propyl, or iso-propyl.

In some embodiments of Formula (III), Ry is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl. In other embodiments, Ry is hydrogen, C1-C3 alkyl, or C1-C3 haloalkyl.

In some embodiments of Formula (III), each Rz is independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl. In other embodiments, Rz is hydrogen, C1-C3 alkyl, or C1-C3 haloalkyl.

In some embodiments of Formula (III), n is 0, 1, 2, or 3. In other embodiments, n is 0 or 1. In further embodiments, n is 0.

In some embodiments of Formula (III), one of Ra and Rb is hydrogen and the other is CO2Rx, CH2CO2Rx, tetrazole, or oxadiazolone. In other embodiments, Rb is hydrogen and Ra is CH2CO2H, tetrazole, or (l,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (III), Rbis hydrogen, Rcis -CN, Rdis thienyl, and Ra is CH2CO2H, tetrazole, or (l,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (III), Rc is halogen, Ra is -CO2H, and Rb is H. In other embodiments, Rc is -Br, Ra is -CO2H, and Rb is H. In further embodiments, Rc is -Cl, Ra is -CO2H, and Rb is H.

In some embodiments of Formula (III), Rc is halogen, Ra is tetrazole, and Rb is H. In other embodiments, Rc is -Br, Ra is tetrazole, and Rb is H. In further embodiments, Rc is -Cl, Ra is tetrazole, and Rb is H.

In some embodiments of Formula (III), Rc is halogen, Ra is -CH2CO2H, and Rb is H. In other embodiments, Rc is -Br, Ra is -CH2CO2H, and Rb is H. In further embodiments, Rc is -Cl, Ra is -CH2CO2H, and Rb is H.

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In some embodiments of Formula (III), Rc is halogen, Ra is (l,2,4-oxadiazol-5(4H)one), and Rb is H. In other embodiments, Rc is -Br, Ra is (l,2,4-oxadiazol-5(4H)-one), and Rb is H. In other embodiments, Rc is -Cl, Ra is (l,2,4-oxadiazol-5(4H)-one), and Rb is H.

In some embodiments of Formula (III), Rc is -CN, Ra is -CO2H, and Rb is H. In other embodiments, Rc is -CN, Ra is -CH2CO2H, and Rb is H. In other embodiments, Rc is -CN, Ra is tetrazole, and Rb is H. In yet other embodiments, Rc is -CN, Ra is (l,2,4-oxadiazol-5(4H)one), and Rb is H.

In some embodiments of Formula (III), Rc is not hydrogen or -CN when Rd is optionally substituted phenyl, Rc is not C1-C6 alkyl when Rd is methyl, and that Rc is not -CN when Rd is 2-furyl.

In some embodiments of Formula (I), (la), (lb), (II) and (III), one of Ra or Rb is a carboxylic acid or a carboxylic acid bioisostere.

In some embodiments of Formula (I), (la), (lb), (II) and (III), Ra is -CO2H, -(CH2)CO2H, or -OCH2CO2H. In other embodiments, Ra is -CO2CH3, -CO2CH2CH3, -CO2CH2CH2CH3, -CO2CH(CH3)2, -(CH2)CO2CH3, -(CH2)CO2CH2CH3, -(CH2)CO2CH2CH2CH3, or -(CH2)CO2CH(CH3)2.

In some embodiments of Formula (I), (la), (lb), (II) and (III), Ra is -P(O)(OH)OH, -(CH2)P(O)(OH)OH, -P(O)(OH)OCH3, -P(O)(OH)OCH2CH3, -P(O)(OH)OCH2CH2CH3, -P(O)(OH)OCH(CH3)2, -(CH2) P(O)(OH)OCH3, -(CH2)P(O)(OH)OCH2CH3, -(CH2)P(O)(OH)OCH2CH2CH3, or -(CH2)P(O)(OH)OCH(CH3)2.

In some embodiments of Formula (I), (la), (lb), (II) and (III), Ra is -S(O)2OH, -(CH2)S(O)2OH, -C(O)NHCN, or -(CH2)C(O)NHCN.

In some embodiments of Formula (I), (la), (lb), (II) and (III), Ra is -C(O)NHS(O)2CH3, -C(O)NHS(O)2CH2CH3, -C(O)NHS(O)2CH2CH2CH3, -C(O)NHS(O)2CH(CH3)2, -(CH2)C(O)NHS(O)2CH3, -(CH2)C(O)NHS(O)2CH2CH3, -(CH2)C(O)NHS(O)2CH2CH2CH3, or -(CH2)C(O)NHS(O)2CH(CH3)2.

In some embodiments of Formula (I), (la), (lb), (II) and (III), Ra is

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OH

In some embodiments of Formula (I), (la), (lb), (II), and (III), Ra is

In some embodiments of Formula (I), (la), (lb), (II) and (III), Rb is -CO2H, -(CH2)CO2H, or -OCH2CO2H. In other embodiments, Rb is -CO2CH3, -CO2CH2CH3, -CO2CH2CH2CH3, -CO2CH(CH3)2, -(CH2)CO2CH3, -(CH2)CO2CH2CH3, -(CH2)CO2CH2CH2CH3, or -(CH2)CO2CH(CH3)2.

In some embodiments of Formula (I), (la), (lb), (II) and (III), Rb is -P(O)(OH)OH, -(CH2)P(O)(OH)OH, -P(O)(OH)OCH3, -P(O)(OH)OCH2CH3, -P(O)(OH)OCH2CH2CH3, -P(O)(OH)OCH(CH3)2, -(CH2) P(O)(OH)OCH3, -(CH2)P(O)(OH)OCH2CH3, -(CH2)P(O)(OH)OCH2CH2CH3, or -(CH2)P(O)(OH)OCH(CH3)2.

In some embodiments of Formula (I), (la), (lb), (II) and (III), Rb is -S(O)2OH, -(CH2)S(O)2OH, -C(O)NHCN, or -(CH2)C(O)NHCN.

In some embodiments of Formula (I), (la), (lb), (II) and (III), Rb is -C(O)NHS(O)2CH3, -C(O)NHS(O)2CH2CH3, -C(O)NHS(O)2CH2CH2CH3, -C(O)NHS(O)2CH(CH3)2, -(CH2)C(O)NHS(O)2CH3, -(CH2)C(O)NHS(O)2CH2CH3, -(CH2)C(O)NHS(O)2CH2CH2CH3, or -(CH2)C(O)NHS(O)2CH(CH3)2.

In some embodiments of Formula (I), (la), (lb), (II) and (III), Rb is

In some embodiments of Formula (I), (la), (lb), (II), and (III), Rb is

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In some embodiments, the compound of Formula (I) is a compound having any one of

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or a pharmaceutically acceptable salt thereof.

The above definition of the compounds of Formula (I) is referred to herein by the expressions “compound of Formula (1)” as defined herein, or simply “compounds of Formula (I)”, etc. The above definition of the compounds of Formula (la) is referred to herein by the expressions “compound of Formula (la)” as defined herein, or simply “compounds of Formula (la)”, etc. The above definition of the compounds of Formula (lb) is referred to herein by the expressions “compound of Formula (lb)” as defined herein, or simply “compounds of Formula (lb)”, etc. The above definition of the compounds of Formula (II) is referred to herein by the expressions “compound of Formula (II)” as defined herein, or simply “compounds of Formula (II)”, etc. The above definition of the compounds of Formula (III) is referred to herein by the expressions “compound of Formula (III)” as defined herein, or simply “compounds of Formula (III)”, etc. It should be understood, that such references are intended to encompass not only the above general formula, but also each and every of the embodiments, etc. discussed in the following. It should also be understood, that unless stated to the opposite, such references also encompass isomers, mixtures of isomers, pharmaceutically acceptable salts, solvates and prodrugs of the compounds of Formula (I), Formula (la), Formula (lb), Formula (II), and Formula (III).

Definitions

The term “alkyl” as used herein refers to a saturated, straight or branched hydrocarbon chain. The hydrocarbon chain preferably contains from one to eight carbon atoms (C!_8alkyl), more preferred from one to six carbon atoms (C!-6-alkyl), in particular from one to four carbon atoms (C!.4-alkyl), including methyl, ethyl, propyl, isopropyl, butyl, isobutyl, secondary butyl, tertiary butyl, pentyl, isopentyl, neopentyl, tertiary pentyl, hexyl, isohexyl, heptyl and octyl. In a preferred embodiment “alkyl” represents a C!.4-alkyl group, which may in particular include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, secondary butyl, and tertiary butyl. Correspondingly, the term “alkylene” means the corresponding biradical (-alkyl-).

The term “cycloalkyl” or “carbocycle” as used herein refers to a cyclic alkyl group, preferably containing from three to ten carbon atoms (C3_10־cycloalkyl or C3.!0-carbocycle),

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The term “alkenyl” as used herein refers to a straight or branched hydrocarbon chain or cyclic hydrocarbons containing one or more double bonds, including di-enes, tri-enes and poly-enes. Typically, the alkenyl group comprises from two to eight carbon atoms (C2-8alkenyl), such as from two to six carbon atoms (C2-6־alkenyl), in particular from two to four carbon atoms (C2-4־alkenyl), including at least one double bond. Examples of alkenyl groups include ethenyl; 1- or 2-propenyl; 1-, 2- or 3-butenyl, or 1,3-but-dienyl; 1-, 2-, 3-, 4- or 5hexenyl, or 1,3-hex-dienyl, or 1,3,5-hex-trienyl; 1-, 2-, 3-, 4-, 5-, 6-, or 7-octenyl, or 1,3octadienyl, or 1,3,5-octatrienyl, or 1,3,5,7-octatetraenyl, or cyclohexenyl. Correspondingly, the term “alkenylene” means the corresponding biradical (-alkenyl-). Alkenyl groups may be optionally substituted with 1-4 substituents. Examples of substituents on alkenyl groups include, but are not limited to, alkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl, hydroxyl, carbamoyl, oxo, and -CN.

The term “alkynyl” as used herein refers to a straight or branched hydrocarbon chain containing one or more triple bonds, including di-ynes, tri-ynes and poly-ynes. Typically, the alkynyl group comprises of from two to eight carbon atoms (C2-8־alkynyl), such as from two to six carbon atoms (C2-6־alkynyl), in particular from two to four carbon atoms (C2-4־alkynyl), including at least one triple bond. Examples of preferred alkynyl groups include ethynyl; 1or 2-propynyl; 1-, 2- or 3-butynyl, or 1,3-but-diynyl; 1-, 2-, 3-, 4- or 5-hexynyl, or 1,3-hexdiynyl, or 1,3,5-hex-triynyl; 1-, 2-, 3-, 4-, 5-, 6-, or 7-octynyl, or 1,3-oct-diynyl, or 1,3,5-octtriynyl, or 1,3,5,7-oct-tetraynyl. Correspondingly, the term “alkynylene” means the corresponding biradical (-alkynyl-). Alkynyl groups may be optionally substituted with 1-4 substituents. Examples of substituents on alkynyl groups include, but are not limited to״ alkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl, hydroxyl, carbamoyl, oxo, and -CN.

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The terms “halo” and “halogen” as used herein refer to fluoro, chloro, bromo or iodo. Thus a trihalomethyl group represents, e.g., a trifluoromethyl group, or a trichloromethyl group. Preferably, the terms “halo” and “halogen” designate fluoro or chloro.

The term “haloalkyl” as used herein refers to an alkyl group, as defined herein, which is substituted one or more times with one or more halogen. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, difluoromethyl, pentafluoroethyl, trichloromethyl, etc.

The term “alkoxy” as used herein refers to an alkyl-O- group, wherein alkyl is as defined above.

The term “hydroxyalkyl” as used herein refers to an alkyl group (as defined hereinabove), which alkyl group is substituted one or more times with hydroxy. Examples of hydroxyalkyl groups include HO-CH2-, HO-CH2-CH2- and CH3-CH(OH)-.

The term “oxy” as used herein refers to an “-O-” group.

The term “oxo” as used herein refers to an “=O” group.

The term “amine” as used herein refers to primary (R-NH2, R H), secondary ((R)2NH, (R)2 ψ H) and tertiary ((R)3-N, R H) amines. A substituted amine is intended to mean an amine where at least one of the hydrogen atoms has been replaced by the substituent.

The term “carbamoyl” as used herein refers to a “H2N(C=0)-” group.

The term aryl, as used herein, unless otherwise indicated, includes carbocyclic aromatic ring systems derived from an aromatic hydrocarbon by removal of a hydrogen atom. Aryl furthermore includes bi-, tri- and polycyclic ring systems. Examples of preferred aryl moieties include phenyl, naphthyl, indenyl, indanyl, fluorenyl, biphenyl, indenyl, naphthyl, anthracenyl, phenanthrenyl, pentalenyl, azulenyl, and biphenylenyl. Preferred aryl is phenyl, naphthyl or indanyl, in particular phenyl, unless otherwise stated. Any aryl used may be optionally substituted. Correspondingly, the term “arylene” means the corresponding biradical (-aryl-). Aryl groups may be optionally substituted with 1-4 substituents. Examples of substituents on aryl groups include, but are not limited to, alkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl, hydroxyl, and -CN.

The term heteroaryl, as used herein, refers to aromatic groups containing one or more heteroatoms selected from O, S, and N, preferably from one to four heteroatoms, and more preferably from one to three heteroatoms. Heteroaryl furthermore includes bi-, tri- and polycyclic groups, wherein at least one ring of the group is aromatic, and at least one of the rings contains a heteroatom selected from O, S, and N. Heteroaryl also include ring systems

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PCT/EP2015/069808 substituted with one or more oxo moieties. Examples of preferred heteroaryl moieties include N-hydroxytetrazolyl, N-hydroxytriazolyl, N-hydroxyimidazolyl, furanyl, triazolyl, pyranyl, thiadiazinyl, benzothiophenyl, dihydro-benzo[b]thiophenyl, xanthenyl, isoindanyl, acridinyl, benzisoxazolyl, quinolinyl, isoquinolinyl, phteridinyl, azepinyl, diazepinyl, imidazolyl, thiazolyl, carbazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl, benzoforanyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, triazinyl, isoindolyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzoforazanyl, benzothiophenyl, benzotriazolyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, dihydroquinolyl, tetrahydroquinolyl, dihydroisoquinolyl, tetrahydroisoquinolyl, benzoforyl, furopyridinyl, pyrolopyrimidinyl, azaindolyl, pyrazolinyl, l,2,4-oxadiazol-5(4H)-one, and pyrazolidinyl. Non-limiting examples of partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl, 1,4-dihydronaphthyl, and 1-octalin. Correspondingly, the term “heteroarylene” means the corresponding biradical (-heteroaryl-). Heteroaryl groups may be optionally substituted with 1-4 substituents. Examples of substituents on heteroaryl groups include, but are not limited to, alkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl, hydroxyl, and -CN.

The term “heterocyclyl” as used herein, refers to cyclic non-aromatic groups containing one or more heteroatoms selected from O, S, and N, preferably from one to four heteroatoms, and more preferably from one to three heteroatoms. Heterocyclyl furthermore includes bi-, tri- and polycyclic non-aromatic groups, and at least one of the rings contains a heteroatom selected from O, S, and N. Heterocyclyl also include ring systems substituted with one or more oxo moieties. Examples of heterocyclic groups are oxetane, pyrrolidinyl, pyrrolyl, 3H-pyrrolyl, oxolanyl, furanyl, thiolanyl, thiophenyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolidinyl, 3H-pyrazolyl, 1,2-oxazolyl, 1,3-oxazolyl, 1,2-thiazolyl, 1,3thiazolyl, 1,2,5-oxadiazolyl, piperidinyl, pyridinyl, oxanyl, 2-H-pyranyl, 4-H-pyranyl, thianyl, 2H-thiopyranyl, pyridazinyl, 1,2-diazinanyl, pyrimidinyl, 1,3-diazinanyl, pyrazinyl, piperazinyl, 1,4-dioxinyl, 1,4-dioxanyl, 1,3-diazinanyl, 1,4-oxazinyl, morpholinyl, thiomorpholinyl, 1,4-oxathianyl, benzofuranyl, isobenzofuranyl, indazolyl, benzimidazolyl, quinolinyl, isoquinolinyl, chromayl, isochromanyl, 4H-chromenyl, IH-isochromenyl, cinnolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, purinyl, naphthyridinyl, pteridinyl, indolizinyl, IH-pyrrolizinyl, 4H-quinolizinyl and aza-8-bicyclo[3.2.1]octane. Correspondingly, the term “heterocyclylene” means the corresponding biradical (-heterocyclyl-). Heterocyclyl groups may be optionally substituted with 1-4 substituents.

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Examples of substituents on heterocyclyl groups include, but are not limited, to alkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl, hydroxyl, and -CN.

The term “N-heterocyclic ring” as used herein, refers to a heterocyclyl or a heteroaryl, as defined hereinabove, having at least one nitrogen atom, and being bound via a nitrogen atom. Examples of such N-heterocyclic rings are pyrrolidinyl, pyrrolyl, 3H-pyrrolyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolidinyl, 3H-pyrazolyl, 1,2-oxazolyl, 1,2thiazolyl, 1,3-thiazolyl, piperidinyl, pyridinyl, pyridazinyl, pyrazinyl, piperazinyl, morpholinyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazolyl, pyrazinyl, tetrazolyl, etc.

Isomers

In the present specification, the structural formula of the compound represents a certain isomer for convenience in some cases, but the present disclosure includes all isomers, such as geometrical isomers, optical isomers based on an asymmetrical carbon, stereoisomers, tautomers, and the like. Accordingly, it should be understood that the definition of compounds of Formulae (I), (la), (lb), (II) and (III) include each and every individual isomer corresponding to the Formula: Formulae (I), (la), (lb), (II) and (III), including cis-trans isomers, stereoisomers and tautomers, as well as racemic mixtures of these and pharmaceutically acceptable salts thereof. Hence, the definition of compounds of Formulae (I), (la), (lb), (II) and (III) are also intended to encompass all R- and S-isomers of a chemical structure in any ratio, e.g., with enrichment (i.e., enantiomeric excess or diastereomeric excess) of one of the possible isomers and corresponding smaller ratios of other isomers. In addition, a crystal polymorphism may be present for the compounds represented by Formulae (I), (la), (lb), (II) and (III). It is noted that any crystal form, crystal form mixture, or anhydride or hydrate thereof is included in the scope of the present disclosure. Furthermore, so-called metabolite which is produced by degradation of the present compound in vivo is included in the scope of the present disclosure.

“Isomerism” means compounds that have identical molecular formulae but differ in the sequence of bonding of their atoms or in the arrangement of their atoms in space. Isomers that differ in the arrangement of their atoms in space are termed “stereoisomers”. Stereoisomers that are not mirror images of one another are termed “diastereoisomers”, and stereoisomers that are non-superimposable mirror images of each other are termed “enantiomers” or sometimes optical isomers. A mixture containing equal amounts of individual enantiomeric forms of opposite chirality is termed a “racemic mixture”.

A carbon atom bonded to four non-identical substituents is termed a “chiral center”.

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Chiral isomer” means a compound with at least one chiral center. Compounds with more than one chiral center may exist either as an individual diastereomer or as a mixture of diastereomers, termed “diastereomeric mixture”. When one chiral center is present, a stereoisomer may be characterized by the absolute configuration (R or S) of that chiral center. Absolute configuration refers to the arrangement in space of the substituents attached to the chiral center. The substituents attached to the chiral center under consideration are ranked in accordance with the Sequence Rule of Cahn, Ingold and Prelog. (Cahn et al., Angew. Chem. Inter. Edit. 1966, 5, 385; errata 511; Cahn et al., Angew. Chem. 1966, 78, 413; Cahn and Ingold, J. Chem. Soc. 1951 (London), 612; Cahn et al., Experientia 1956, 12, 81; Cahn, J. Chem. Educ. 1964, 41, 116).

Diastereoisomers, i.e., non-superimposable stereochemical isomers, can be separated by conventional means such as chromatography, distillation, crystallization or sublimation. The optical isomers can be obtained by resolution of the racemic mixtures according to conventional processes, for example by formation of diastereoisomeric salts by treatment with an optically active acid or base. Examples of appropriate acids include, without limitation, tartaric, diacetyltartaric, dibenzoyltartaric, ditoluoyltartaric and camphorsulfonic acid. The mixture of diastereomers can be separated by crystallization followed by liberation of the optically active bases from these salts. An alternative process for separation of optical isomers includes the use of a chiral chromatography column optimally chosen to maximize the separation of the enantiomers. Still another available method involves synthesis of covalent diastereoisomeric molecules by reacting compounds of Formula (I), (la), (lb), (II) or (III) with an optically pure acid in an activated form or an optically pure isocyanate. The synthesized diastereoisomers can be separated by conventional means such as chromatography, distillation, crystallization or sublimation, and then hydrolyzed to obtain the enantiomerically pure compound. The optically active compounds of Formulae (I), (la), (lb), (II) and (III) can likewise be obtained by utilizing optically active starting materials and/or by utilizing a chiral catalyst. These isomers may be in the form of a free acid, a free base, an ester or a salt. Examples of chiral separation techniques are given in Chiral Separation Techniques, A Practical Approach, 2nd ed. by G. Subramanian, Wiley-VCH, 2001.

“Geometric isomer” means the diastereomers that owe their existence to hindered rotation about double bonds. These configurations are differentiated in their names by the prefixes cis and trans, or Z and E, which indicate that the groups are on the same or opposite side of the double bond in the molecule according to the Cahn-Ingold-Prelog rules.

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Furthermore, the structures and other compounds discussed in this disclosure include all atropic isomers thereof. “Atropic isomers” are a type of stereoisomer in which the atoms of two isomers are arranged differently in space. Atropic isomers owe their existence to a restricted rotation caused by hindrance of rotation of large groups about a central bond. Such atropic isomers typically exist as a mixture, however as a result of recent advances in chromatography techniques; it has been possible to separate mixtures of two atropic isomers in select cases.

“Tautomer” is one of two or more structural isomers that exist in equilibrium and is readily converted from one isomeric form to another. This conversion results in the formal migration of a hydrogen atom accompanied by a switch of adjacent conjugated double bonds. Tautomers exist as a mixture of a tautomeric set in solution. In solid form, usually one tautomer predominates. In solutions where tautomerization is possible, a chemical equilibrium of the tautomers will be reached. The exact ratio of the tautomers depends on several factors, including temperature, solvent and pH. The concept of tautomers that are interconvertible by tautomerizations is called tautomerism.

Of the various types of tautomerism that are possible, two are commonly observed. In keto-enol tautomerism a simultaneous shift of electrons and a hydrogen atom occurs. Ringchain tautomerism arises as a result of the aldehyde group (-CHO) in a sugar chain molecule reacting with one of the hydroxy groups (-OH) in the same molecule to give it a cyclic (ringshaped) form as exhibited by glucose.

Common tautomeric pairs are: ketone-enol, amide-nitrile, lactam-lactim, amideimidic acid tautomerism in heterocyclic rings (e.g., in nucleobases such as guanine, thymine and cytosine), amine-enamine and enamine-enamine. It is to be understood that the compounds of the present disclosure may be depicted as different tautomers. It should also be understood that when compounds have tautomeric forms, all tautomeric forms are intended to be included in the scope of the present disclosure, and the naming of the compounds does not exclude any tautomer form.

The term “crystal polymorphs”, “polymorphs” or “crystal forms” means crystal structures in which a compound (or a salt or solvate thereof) can crystallize in different crystal packing arrangements, all of which have the same elemental composition. Different crystal forms usually have different X-ray diffraction patterns, infrared spectral, melting points, density hardness, crystal shape, optical and electrical properties, stability and solubility. Recrystallization solvent, rate of crystallization, storage temperature, and other

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PCT/EP2015/069808 factors may cause one crystal form to dominate. Crystal polymorphs of the compounds can be prepared by crystallization under different conditions.

Additionally, the compounds of the present disclosure, for example, the salts of the compounds, can exist in either hydrated or unhydrated (the anhydrous) form or as solvates with other solvent molecules. Nonlimiting examples of hydrates include monohydrates, dihydrates, etc. Nonlimiting examples of solvates include ethanol solvates, acetone solvates, etc.

“Solvate” means solvent addition forms that contain either stoichiometric or nonstoichiometric amounts of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming a solvate. If the solvent is water the solvate formed is a hydrate; and if the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one molecule of the substance in which the water retains its molecular state as H2O.

The present disclosure is intended to include all isotopes of atoms occurring in the present compounds. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include C-13 and C-14.

Throughout the description and claims of this specification, the words “comprise” and “contain” and variations of the words, for example “comprising” and “comprises”, mean “including but not limited to” and do not exclude other moieties, additives, components, integers or steps. Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

All references, including any patent or patent application, cited in this specification are hereby incorporated by reference. No admission is made that any reference constitutes prior art. Further, no admission is made that any of the prior art constitutes part of the common general knowledge in the art.

Method of treatment

In another aspect, the present disclosure relates to a method of treating a disease or disorder in which a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) plays a role comprising administering to the subject in need thereof a therapeutically effective

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PCT/EP2015/069808 amount of one or more compounds of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof.

PCT/EP2015/069808 medicament comprises a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a pharmaceutical composition for use in a method for treating a disease or condition mediated by ACMSD, wherein the medicament comprises a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a pharmaceutical composition for use in a method for preventing a disease or condition mediated by ACMSD, wherein the medicament comprises a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use as a medicament for treating, preventing or reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

Another aspect of the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use as a medicament for treating a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

In another aspect, the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use as a medicament for preventing a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

Another aspect of the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use as a medicament for reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

In another aspect, the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use as a medicament for ameliorating a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

Another aspect of the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use as a medicament for treating, preventing or reducing the risk of a disorder associated with mitochondrial dysfunction.

In another aspect, the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use as a medicament for treating a disorder associated with mitochondrial dysfunction.

Another aspect of the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use as a medicament for preventing a disorder associated with mitochondrial dysfunction.

In another aspect, the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable

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PCT/EP2015/069808 salt thereof for use as a medicament for reducing the risk of a disorder associated with mitochondrial dysfunction.

Another aspect of the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use as a medicament for ameliorating a disorder associated with mitochondrial dysfunction.

Another aspect of the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use in treating, preventing or reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

In another aspect, the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use in treating a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

Another aspect of the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use in preventing a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

In another aspect, the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use in reducing the risk of a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

Another aspect of the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use in ameliorating a disease or disorder associated with reduced nicotinamide adenine dinucleotide (NAD+) levels.

In another aspect, the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use in for treating, preventing or reducing the risk of a disorder associated with mitochondrial dysfunction.

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In another aspect, the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use in for preventing a disorder associated with mitochondrial dysfunction.

Another aspect of the present disclosure relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof for use in for reducing the risk of a disorder associated with mitochondrial dysfunction.

O O

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or a pharmaceutically acceptable salt thereof for use in treating, preventing, ameliorating or reducing the risk of a disease or disorder associated with a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction.

In another aspect, the present disclosure relates to a compound having the one of the following Formula:

O O

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associated with a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction.

As used herein, “treating” or “treat” describes the management and care of a patient for the purpose of reversing, inhibiting, or combating a disease, condition, or disorder and includes the administration of a compound of the present disclosure (i.e., a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III)), or a pharmaceutically acceptable salt, prodrug, metabolite, polymorph or solvate thereof, to reverse the disease, condition, or disorder, eliminate the disease, condition, or disorder, or inhibit the process of the disease, condition, or disorder.

A compound of the present disclosure (i.e., a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III)), or a pharmaceutically acceptable salt, prodrug,

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PCT/EP2015/069808 metabolite, polymorph or solvate thereof, can also be used to prevent a disease, condition, or disorder or one or more symptoms of such disease, condition, or disorder. As used herein, “preventing” or “prevent” describes reducing or eliminating the onset of the symptoms or complications of the disease, condition, or disorder.

A compound of the present disclosure (i.e., a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III)), or a pharmaceutically acceptable salt, prodrug, metabolite, polymorph or solvate thereof, can also be used to alleviate one or more symptoms of such disease, condition, or disorder. As used herein, the term alleviate is meant to describe a process by which the severity of a sign or symptom of a disorder is decreased. Importantly, a sign or symptom can be alleviated without being eliminated. Preferably treatment is curative or ameliorating.

Methods for the preparation of compounds of Formulae (I), (la), (lb), (II) and (III)

The compounds of the present disclosure (e.g., compounds of Formula (I), Formula (la), Formula (lb), Formula (II), and Formula (II)) can be prepared in a number of ways well known to those skilled in the art of organic synthesis. By way of example, compounds of the present disclosure can be synthesized using the methods described below, together with synthetic methods known in the art of synthetic organic chemistry, or variations thereon as appreciated by those skilled in the art. Preferred methods include but are not limited to those methods described below. The final products of the reactions described herein may be isolated by conventional techniques, e.g., by extraction, crystallisation, distillation, chromatography, etc.

Compounds of the present disclosure can be synthesized by following the steps outlined in General Scheme A to E which comprise different sequences of assembling intermediates la-Ih and Ij-Io. Starting materials are either commercially available or made by known procedures in the reported literature or as illustrated. Useful steps that may be used in the preparation steps of the compounds will be known to the skilled person. The method below is given as a non-limiting example on how the compounds may be prepared.

General Scheme A

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PCT/EP2015/069808 wherein R1, Rc, Rd, and L are defined as in Formula (I).

The general way of preparing compounds of Formula (I) by using intermediates la, and lb is outlined in General Scheme A. Coupling of la with lb using a base, i.e., potassium carbonate (K2CO3), in a solvent, i.e., acetonitrile (CH3CN), optionally at elevated temperature provides the desired produce of Formula (I). Bases that can be used include, but are not limited to, sodium carbonate (Na2CO3), potassium carbonate (K2CO3), N,Ndiisopropylethylamine (DIPEA) and triethylamine. Solvents used in the coupling reaction can be polar or non-polar solvents. For example, the solvent can be acetonitrile (CH3CN), acetone, or dimethylsulfoxide (DMSO).

General Scheme B

(I)

Ic Id wherein X is a good leaving group, i.e., Cl, Br, -SCH3, or S(O)2CH3, and R1, R2, Rc, Rd, and p are defined as in Formula (I).

Alternatively, compounds of Formula (I) can be prepared using intermediates Ic and Id as outlined in General Scheme B. Amination of Intermediate Ic with Ie using a base, i.e., sodium hydroxide (NaOH), potassium hydroxide (KOH), etc., in a solvent, i.e., methanol (MeOH), ethanol (EtOH), water (H2O), etc., provides compounds of Formula (I).

General Scheme C

O

O

(I)

Ie If wherein X is a good leaving group, i.e., Cl, Br, -SCH3, or S(O)2CH3, and R1, R2, Rc, Rd, and p are defined as in Formula (I).

Compounds of Formula (I) can also be prepared using intermediates Ie and If as outlined in General Scheme C. Amination of Intermediate Ie with If using a base, i.e.,

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PCT/EP2015/069808 sodium hydroxide (NaOH), potassium hydroxide (KOH), etc., in a solvent, i.e., methanol (MeOH), ethanol (EtOH), water (H2O), etc., provides compounds of Formula (I).

General Scheme D

Ih Ij

Ik Im I wherein and R1, Rc, and Rd are defined as in Formula (I).

Alternatively, compounds of Formula (I) can also be prepared using intermediates Ig, Ih, Ij, Ik, and Im as outlined in General Scheme D. Olefination of intermediate Ig using a base i.e., potassium carbonate (K2CO3) and diethyl (cyanomethyl)phosphonate in a solvent, i.e., tetrahydrofuran (THF), water (H2O), optionally at an elevated temperature provides Intermediate Ih. Hydrogenation of Ih using a metal catalyst, i.e., palladium on carbon (Pd/C), platinum dioxide (PtO2), etc, and hydrogen (H2) gas in a solvent, i.e., ethanol (EtOH) and/ or tetrahydrofuran (THF), provides Intermediate Ij. Intermediate Ik is obtained by treating Intermediate Ij with an acid, i.e., hydrochloric acid (HC1) in a solvent, i.e., ethanol (EtOH), dichloromethane (CH2C12), etc., and then subsequent treatment with a base, i.e., ammonia (NH3). Cyclization of Intermediate Ik and Im using a base, i.e., sodium hydroxide (NaOH), potassium hydroxide (KOH), etc., in a solvent, i.e., dimethylacetamide (DMA), optionally at elevated temperature provides compounds of Formula (I).

General Scheme E

O

wherein and R1, Rc, and Rd are defined as in Formula (I).

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Alternatively, compounds of Formula (I) can be prepared using intermediates In and 10 as outlined in General Scheme D. Acylation of Intermediate In with 10 using a base, i.e., sodium hydroxide (NaOH), potassium hydroxide (KOH), etc., in a solvent, i.e., methanol (MeOH), ethanol (EtOH), water (H2O), etc., provides compounds of Formula (I).

A mixture of enantiomers, diastereomers, cis/trans isomers resulting from the process described above can be separated into their single components by chiral salt technique, chromatography using normal phase, reverse phase or chiral column, depending on the nature of the separation.

It should be understood that in the description and formula shown above, the various groups R1, R2, X, L, Y, Ra, Rb, Rc, Rd, Re, Rf, Rx, Ry, Rz, m, n, p, q, r and other variables are as defined herein above, except where otherwise indicated. Furthermore, for synthetic purposes, the compounds of General Schemes 1 and 2 are mere representative with elected radicals to illustrate the general synthetic methodology of the compounds of Formula (I) as defined herein.

Biological Assays and Animals Studies

Method of Screening ACMSD1 Inhibition

The activity of compounds as inhibitors of ACMSD1 is determined in a spectrophotometrical in vitro assay. The pre-assay mixture is incubated and a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, and ACMSD 1 solution is then added. The effect of ACMS concentration on the enzyme activity is investigated by varying 3-hydroxyanthranilic acid (30H-HA) concentration in the pre-assay mixture. Kinetic parameters are calculated from the initial velocity data using a Lineweaver-Burk plot.

Cellular Assay Methods

The mouse hepatocytes cell lines are grown and plated. The cells are maintained in culture at 37 °C and once the cells are attached, different concentrations of a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, or DMSO are added. Primary hepatocytes are harvested about 24 hrs later.

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Determination of ACMSD-1 Modulation in HEK293T Cells.

HEK293T cells are seeded and transfected to transiently express ACMSD. The cells are then stimulated with different concentrations of Compound 1, and then lysed to measure the ACMSD activity in a spectrophotometrical in vitro assay. The amount of the whole protein content in cell lysates is detected by Bradford analysis and used to get the specificity activity of the enzyme normalized in all samples.

Determination of NAD+ content in Human Primary Hepatocytes

Primary hepatocytes are treated with different concentrations of a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, or MEHP (control) after seeding. The compound is replaced every 24 hours, and then cells are directly harvested and lysed to detect NAD+ content through LC MS/MS (liquid chromatography mass spectrometry/mass spectroscopy).

Modulation ofSOD2 activity in AML12 cells and Murine Primary Hepatocytes

Primary hepatocytes or AML-12 cells are lysed and total protein concentration is determined using the Bradford assay. SOD2 activity is determined at indicated times after treatment with a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, using a SOD Assay Kit. Absorbance is determined and results are expressed in U/ml/mg of protein according to the standard curve and measured protein concentration.

Determination of NAD+ content in Murine Primary Hepatocytes

NAD+ is extracted using acidic extraction method and samples are collected and homogenized. After insoluble protein parts are pelleted, the samples are separated by highperformance liquid chromatography (HPLC) and analyzed by mass-spectrometry. The proteins in the pellet are quantified by Bradford assay and are used for normalization.

RNA preparation and RT-qPCR analysis of ACMSD and SIRT1 -regulated Genes in Cells,

Cells (AML-12, Hepa-1.6, HEK-293, primary human and murine hepatocytes) are treated with different concentrations of a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof and the gene expression of ACMSD, Pgcla, Sodl, and Sod2 (MnSOD) is determined using RT-qPCR. Total RNA is extracted from cells and the extracted RNA is treated with DNase and used for reverse transcription (RT).

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Modulation of Caspase 3/7 Activity in MDCK Cells

MDCK cells are cultured in base medium to a final concentration of 10%. Cells are plated into 96 wells and 24 hours after cell plating the medium is changed with fresh medium supplemented with 1% FBS. Cisplatin is then used to induce cell injury. Different concentrations of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof (in DMSO) are added in combination with cisplatin or prior to adding cisplatin. Caspase 3/7 activity (Promega) is determined according to standard procedures using a luminescent signal readout on a plate reader. Each experiment/condition is performed in triplicate. Caspase activity is analyzed as percentage effect normalized to the cisplatin alone and vehicle treated cells.

Cytotoxicity and hERG screening

HePG2 and AML-12 cells are seeded and a dose-response of the compound is performed at various concentrations. Cells are stimulated and the supernatant is used to perform LDH release as a measure of necrosis while the cells are lysed to detect ATP levels for determining cell viability.

The Predictor hERG assay kit is stably transfected with hERG potassium channel and a high-affinity red fluorescent hERG channel ligand and is used for the determination of hERG channel affinity binding of compounds of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof. Compounds that bind to the hERG channel protein (competitors) are identified by their ability to displace the tracer which results in a lower fluorescence polarization.

C.elegans experiments - ACMSD1 silencing, lifespan assays, mobility assessment and GFP quantification

ACMSD1 silencing: Bacterial feeding RNAi experiments to determine the effects of downregulation or silencing of acmsd-1 on gene expression and survival are carried out in the nematode Caenorhabditis elegans (C. elegans). The clones used for the bacterial feeding experiments are acmsd-1, SIR-2.1 and DAF-16. Total RNA is extracted from cells and the extracted RNA is treated with DNase, and used for reverse transcription (RT).

Worms are grown on NGM agar plates additionally containing Carbenicillin and IPTG and seeded with bacterial cultures. After RNAi treatment, worms are transferred to plates containing paraquat and seeded with RNAi bacteria. Control animals are grown on RNAi bacteria containing an empty vector (control) and then transferred to plates containing

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PCT/EP2015/069808 paraquat and seeded with RNAi bacteria. Quantification of gene expression of sod-3 at mRNA levels and protein levels using RT-qPCR and survival analyses are performed. The movement of worms is recorded at days 1,3, and 5 of adulthood.

Anti-diabetic Effects studies in C57BL/6J and KK-Ay mice

Mice are fed with regular chow or a high fat diet (HFD). A compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, is dosed daily and blood and tissues are harvested for RNA isolation, lipid measurements and histology. Oxygen consumption is measured and histological analysis and transmission electron microscopy are performed. An oral glucose tolerance test and an intraperitoneal insulin tolerance test are also performed to quantify glucose and to measure plasma insulin concentrations.

Anti-diabetic and Anti-obesity studies in db/db Mice with LepR Mutation

Animals are fed a high-fat diet (HFD). For subchronic intervention, the animals are treated once/day with a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, for 14 days. Blood samples are collected and glucose concentrations of each blood sample are determined. For acute intervention, initial blood samples are collected and then compounds of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, are administered. Diet-access is then restricted, and a second blood sample is collected. The mice are subjected to an oral glucose tolerance test and blood glucose concentrations are determined.

For the euglycemic-hyperinsulinemic clamps assay, the animals receive a primedcontinuous [3-3H]glucose infusion and a blood sample is then collected to determine plasma insulin, glucose and [3-3H]glucose concentrations and to calculate basal endogenous glucose appearance rates. The mice then receive vehicle or a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, via gavage. Subsequently, the animals receive a [3-3H]glucose infusion containing insulin causing a moderate net-increase in plasma insulin concentrations. Blood glucose concentrations are measured and target glycemia is established by adjusting the rate of glucose infusion. 2-deoxy-D-[l-14C] glucose is then given intravenously and blood samples are collected. The mice are then sacrificed. Gastrocnemius muscle and epididymal adipose

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PCT/EP2015/069808 tissue are collected and plasma [3H]- and [14C]-radioactivity is determined in deproteinized plasma.

Body weights are assessed and brown adipose tissue (BAT) and gonadal white adipose tissue (WAT) are dissected and weighed. Volume oxygen (VO2) and volume carbon dioxide production (VCO2) are measured and are reported as average VO2 per hour normalized to body weight (mL/h/kg). Activity counts by infrared beam interruptions and food intake are simultaneously measured.

Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) Studies in Male C57BL/6J mice

Mice are fed a ‘Western’ HF-HSD (high fat-high sucrose diet) or normal chow diet (NCD) as control. The animals are then treated with a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, for 4, 12 or 20 weeks, and then sacrificed. Body weight and food intake are monitored weekly and total fat mass is analysed. An intraperitoneal glucose tolerance test (IPGTT) is also performed and tail vein glucose levels are measured after glucose administration. Insulin resistance is calculated using the Homeostasis Model of Insulin Resistance. The mice are then sacrificed by blood sampling via cardiac puncture. Plasma is obtained and tissues were collected together with the plasma for further biochemical and molecular analyses or for histological analysis.

Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) Studies in in Methionine and Choline Deficient mice

Mice weighing 25 g are either fed a methionine- and choline-deficient diet (MCD to induce NASH) or chow diet (as a control). Animal experiments and evaluation of NAFLD and NASH are conducted as described above in for C57BL/6J mice fed the high fat and high sucrose diet.

Atherosclerosis Studies in High Cholesterol Fed LDL-R Knockout mice

LDL-R knockout (KO) mice are sacrificed about 12 weeks after the initiation of the atherogenic diet, after which the heart and aorta are perfused with PBS and subsequently fixed. Atherosclerosis and biochemistry parameters are measured with the appropriate commercially available kits. For the in vivo lipopolysaccharide (LPS) study, mice are

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PCT/EP2015/069808 intraperitoneally injected with LPS, and blood is taken from the tail vein. TNFa levels are quantified with a Mouse TNFa ELISA assay. Blood cell counts are determined.

Dosage and administration are adjusted to provide sufficient levels of the active agent(s) or to maintain the desired effect. Factors which may be taken into account include the severity of the disease state, general health of the subject, age, weight, and gender of the subject, diet, time and frequency of administration, drug combination(s), reaction sensitivities, and tolerance/response to therapy. Long-acting pharmaceutical compositions may be administered every 3 to 4 days, every week, or once every two weeks depending on half-life and clearance rate of the particular formulation.

A suitable dosage of the compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, will depend on the age and condition of the patient, the severity of the disease to be treated and other factors well known to the practicing physician. The compound may be administered for example either orally, parenterally or topically according to different dosing schedules, e.g., daily or with intervals, such as weekly intervals. In general a single dose will be in the range from 0.01 to 500 mg/kg body weight, preferably from about 0.05 to 100 mg/kg body weight, more preferably between 0.1 to 50 mg/kg body weight, and most preferably between 0.1 to 25 mg/kg body weight. The compound may be administered as a bolus (i.e., the entire daily dose is administered at once) or in divided doses two or more times a day. Variations based on the aforementioned dosage ranges may be made by a physician of ordinary skill taking into account known considerations such as weight, age, and condition of the person being treated, the severity of the affliction, and the particular route of administration.

As used herein, a “subject” or “subject in need thereof’ is a subject having a disease or disorder associated with a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) dysfunction. A “subject” includes a mammal. The mammal can be e.g., any

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PCT/EP2015/069808 mammal, e.g., a human, primate, bird, mouse, rat, fowl, dog, cat, cow, horse, goat, camel, sheep or a pig. Preferably, the mammal is a human.

The compounds of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, may also be prepared in a pharmaceutical composition comprising one or more further active substances alone, or in combination with pharmaceutically acceptable carriers, diluents, or excipients in either single or multiple doses. The suitable pharmaceutically acceptable carriers, diluents and excipients are as described herein above, and the one or more further active substances may be any active substances, or preferably an active substance as described in the section “combination treatment” herein below.

Clinical conditions and other uses of compounds

The compounds according to Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable form thereof, compositions, medicaments, and compounds for use, as defined herein, are useful for treatment of a disease or disorder in which a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) modulation plays a role. The compounds may be used either in human or in veterinary medicine and the patient may be any mammal, but especially a human. The treatment may include administering to any mammal, but especially a human, suffering from a disease or disorder in which a-amino-3־carboxymuconate-8-semialdehyde decarboxylase (ACMSD) modulation plays a role, a therapeutically effective amount of a compound according to Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, as defined herein.

The present disclosure also relates to a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, as defined herein, for use in a disease or disorder associated with a-amino-3־carboxymuconateε-semialdehyde decarboxylase (ACMSD) dysfunction, such as obesity, type II diabetes and its complications (e.g., diabetic retinopathy and nephropathy), non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or chronic kidney disease.

By the term “disease or disorder associated with a-amino-3־carboxymuconate-8semialdehyde decarboxylase (ACMSD) dysfunction” is meant any disease characterized by reduced nicotinamide adenine dinucleotide (NAD+) expression and/or activity in at least in some instances of the disease, or a disease which is ameliorated by elevation of the levels of NAD+.

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The methods, medicaments and compounds for use of the present disclosure are useful to treat, alleviate the symptoms of, or delay the onset of a disorder associated with aberrant mitochondrial function. Disorders associated with aberrant mitochondrial function include, for example, metabolic disorders, neurodegenerative disorders, aging related disorders, and chronic inflammatory disorders. Mitochondrial disorders also include diseases with inherited and/or acquired mitochondrial dysfunction (i.e., Charcot-Marie-Tooth disease, Type 2A2, Mitochondrial Encephalopathy Lactic Acidosis and Stroke (MELAS), Leigh syndrome, Barth syndrome, and Leber’s optic neuropathy), fatty acid oxidation disorders, inherited forms of deafness and blindness, and metabolic abnormalities induced by exposure to toxic chemicals and/or drugs (e.g., cisplatin induced deafness).

Metabolic disorders include, for example, type II diabetes, obesity, hyperglycemia, glucose intolerance, insulin resistance (i.e., hyperinsulinemia, metabolic syndrome, syndrome X), hypercholesterolemia, hypertension, hyperlipoproteinemia, hyperlipidemia (e.g., dyslipidemia), hypertriglylceridemia, cardiovascular disease, atherosclerosis, peripheral vascular disease, kidney disease, ketoacidosis, thrombotic disorders, nephropathy, diabetic neuropathy, diabetic retinopathy, sexual dysfunction, dermatopathy, dyspepsia, hypoglycemia, cancer, and edema.

Neurodegenerative disorders include diseases such as photoreceptor degeneration (i.e., retinitis pigmentosa), Dementia, Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease.

Chronic inflammatory diseases include diseases such as celiac disease, vasculitis, lupus, chronic obstructive pulmonary disease (COPD), irritable bowel disease, atherosclerosis, arthritis, and psoriasis.

Aging related disorders include diseases such as cancer, dementia, cardiovascular disease (i.e., arteriosclerosis), hypertension, diabetes mellitus (type I or type II), arthritis, cataracts, Alzheimer's disease, macular degeneration, and osteoporosis.

The subject can be suffering from or susceptible to developing a metabolic disorder. Subjects suffering from or at risk of developing a metabolic disorder are identified by methods known in the art. For example, diabetes can be diagnosed by measuring fasting blood glucose levels or insulin or by glucose tolerance test. Normal adult glucose levels are between about 60-126 mg/dl. Normal insulin levels are about 7 mU/mL + 3mU. Hypertension can be diagnosed by a blood pressure reading consistently at or above about 140/90. Cardiovascular disease can be diagnosed by measuring cholesterol levels. For example, LDL cholesterol above about 137 or total cholesterol above about 200 is indicative

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PCT/EP2015/069808 of cardiovascular disease. Hyperglycemia can be diagnosed by a blood glucose level higher than about 10 mmol/1 (180 mg/dl). Glucose intolerance can be diagnosed by glucose levels of 140 to 199 mg per dL (7.8 to 11.0 mmol) after conducting a 75 g oral two-hour glucose tolerance test. Insulin resistance can be diagnosed by a fasting serum insulin level of greater than approximately 60 pmol/L. Hypoglycemia can be diagnosed by a blood glucose level lower than about 2.8 to 3.0 mmol/L (50 to 54 mg/dl). Obesity can be diagnosed, for example, by body mass index. Body mass index (BMI) is measured in kg/m2 (or lb/in2 X 704.5). Alternatively, waist circumference (estimates fat distribution), waist-to-hip ratio (estimates fat distribution), skinfold thickness (if measured at several sites, estimates fat distribution), or bioimpedance (based on principle that lean mass conducts current better than fat mass (i.e., fat mass impedes current), estimates % fat) can be measured. The parameters for normal, overweight, or obese individuals are as follows: Underweight: BMI < 18.5; Normal: BMI about 18.5 to about 24.9; Overweight: BMI = about 25 to about 29.9. Overweight individuals are characterized as having a waist circumference of > 94 cm for men or > 80 cm for women and waist to hip ratios of > 0.95 in men and > 0.80 in women. Obese individuals are characterized as having a BMI of 30 to 34.9, being greater than 20% above normal weight for height, having a body fat percentage > 30% for women and 25% for men, and having a waist circumference >102 cm (40 inches) for men or 88 cm (35 inches) for women. Individuals with severe or morbid obesity are characterized as having a BMI of > 35.

The methods described herein may lead to a reduction in the severity or the alleviation of one or more symptoms of a metabolic disorder. For example, symptoms of diabetes include elevated fasting blood glucose levels, blood pressure at or above 140/90 mm/Hg; abnormal blood fat levels, such as high-density lipoproteins (HDL) less than or equal to 35 mg/dL, or triglycerides greater than or equal to 250 mg/dL (mg/dL = milligrams of glucose per deciliter of blood). Efficacy of treatment is determined in association with any known method for diagnosing the metabolic disorder. Alleviation of one or more symptoms of the metabolic disorder indicates that the compound confers a clinical benefit.

The methods of the present disclosure are useful to treat, alleviate the symptoms of, or delay the onset of a kidney disorder. Kidney disorders include acute kidney injury (AKI) and chronic kidney disease (CKD).

The subject can be suffering from or susceptible to developing acute kidney injury (AKI). The acute kidney injury can be characterized by one or more clinical criteria or conditions (i.e., an abrupt decrease in the ability of the kidneys to excrete nitrogenous waste products from the blood, resulting in azotemia). Subjects suffering from or at risk of

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PCT/EP2015/069808 developing acute kidney injury (AKI) are identified by methods known in the art. For example, the acute kidney injury can be characterized by an increase in serum creatinine by at least 50% over baseline, an absolute increase in serum creatinine of at least 0.3 mg/dL over baseline, a reduction in glomerular filtration rate of at least 25% compared to baseline, a decrease in urine output to 0.5 ml per kilogram of body weight or less per hour persisting for at least 6 hours, or any combination thereof. An acute kidney injury may be caused by ischemia, drugs or toxic agents (i.e., radiocontrast media, a non-steroidal anti-inflammatory drug (NSAID), alcohol, or a chemotherapy agent), viruses, and obstruction.

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WO 03/000250; WO 03/002530; WO 03/002531; WO 03/002553; WO 03/002593; WO 03/000180; and WO 03/000181.

Sulfonylureas useful in the present disclosure include, but are not limited to, acetohexamide, chlorpropamide, diabinese, glibenclamide, glipizide, glyburide, glimepiride, gliclazide, glipentide, gliquidone, glisolamide, tolazamide, and tolbutamide, pharmaceutically acceptable salts or esters thereof. Meglitinides useful in the present disclosure include, but are not limited to, repaglinide and nateglinide, and pharmaceutically acceptable salts or esters thereof.

Alpha glucoside hydrolase inhibitors (or glucoside inhibitors) useful in the present disclosure include, but are not limited to, acarbose, adiposine, camiglibose, emiglitate, miglitol, voglibose, pradimicin-Q, salbostatin, CKD-711, MDL-25,637, MDL-73,945, and MOR 14, and pharmaceutically acceptable salts or esters thereof, and the compounds disclosed in U.S. Pat. Nos. 4,062,950, 4,174,439, 4,254,256, 4,701,559, 4,639,436, 5,192,772, 4,634,765, 5,157,116, 5,504,078, 5,091,418, 5,217,877, and 5,091,524. Alphaamylase inhibitors useful in the present disclosure include, but are not limited to, tendamistat, trestatin, and Al-3688, and pharmaceutically acceptable salts and esters thereof, and the compounds disclosed in U.S. Pat. Nos. 4,451,455, 4,623,714, and 4,273,765.

Insulin secreatagogues useful in the present disclosure include, but are not limited to, linogliride and A-4166, and pharmaceutically acceptable salts and esters thereof.

Fatty acid oxidation inhibitors useful in the present disclosure include, but are not limited to, clomoxir, and etomoxir, and pharmaceutically acceptable salts and esters thereof. A2 antagonists useful in the present disclosure include, but are not 'limited to, midaglizole, isaglidole, deriglidole, idazoxan, earoxan, fluparoxan, and pharmaceutically acceptable salts and esters thereof. Insulin or insulin mimetics useful in the present disclosure include, but are not limited to, biota, LP-100, novarapid, insulin detemir, insulin lispro, insulin glargine, insulin zinc suspension (lente and ultralente), Lys-Pro insulin, GLP-1 (73-7) (insulintropin), and GLP-1 (7-36)-NH2), and pharmaceutically acceptable salts or esters thereof.

Glycogen phosphorylase inhibitors useful in the present disclosure include, but are not limited to, CP-368, 296, CP-316,819, BAYR3401, and compounds disclosed in WO 01/94300, and WO 02/20530, and pharmaceutically acceptable salts or esters thereof. GLP-1 agonists useful in the present disclosure include, but are not limited to, exendin-3 and exendin-4, and compounds disclosed in US 2003087821 and NZ 504256, and pharmaceutically acceptable salts or esters thereof.

Examples of HMG-C0A reductase inhibitors include, but are not limited to, lovastatin, simvastatin, pravastatin and fluvastatin. Examples of HMG-C0A synthase inhibitors are the beta-lactone derivatives disclosed in U.S. Patents 4,806,564, 4,816,477,

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4,847,271, and 4,751,237; the beta-lactam derivatives disclosed in U.S. 4,983,597 and U.S.S.N. 07/540,992 filed June 20, 1990; and the substituted oxacyclopropane analogues disclosed in European Patent Publication EP 0 411 703. Examples of squalene epoxidase inhibitors are disclosed in European Patent Publication EP 0 318 860 and in Japanese Patent Publication J02 169-571 A. Examples of LDL-receptor gene inducer molecules are disclosed in U.S. Patent 5,182,298 filed March 18, 1991. Other cholesterol lowering agents that may be administered include niacin, probucol, fibric acids (i.e., clofibrate and gemfibrozil), and LDL-receptor gene inducers.

Examples of PARP inhibitors include, but are not limited to, iodonitocoumarin, 5iodo-6-nitrocoumarin, 3,4-dihydro-5-methyl-isoquinolinone, 4-amino-l,8-naphthalimide, 3methoxybenzamide, 8-hydroxy-2-methyl-3- hydro-quinazolin-4-one, 2-{3-[4-(4fluorophenyl)-3,6-dihydro-l(2h)-pyridinyl]propyl}-8-methyl-4(3h)- quinazolinone, 5-fluorol-[4-(4-phenyl-3,6-dihydropyridin-l(butyl]quinazoline-2,4(lh,3h)-dione, 3-(4-chlorophenyl) quinoxaline-5-carboxamide, 2-(3'-methoxyphenyl)benzirnidazole-4-carboxam, benzamide, 3aminobenzamide, 3-aminophtalhydrazide, and 1,5-dihydroxyisoquinoline.

The above-mentioned compounds, which can be used in combination with a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, can be prepared and administered as described in the art such as in the documents cited above.

The above compounds are only illustrative of the ACMSD inhibitors, anti-diabetic agents, anti-obesity agents, cholesterol lower agent, compounds that boost NAD levels, compounds that inhibit NAD consumption that can be used in the compositions of the present disclosure. As this listing of compounds is not meant to be comprehensive, the methods of the present disclosure may employ any anti-obesity agent and any anti-diabetic agent, and are not limited to any particular structural class of compounds.

As used herein, “combination therapy” includes the administration of a compound of the present disclosure, or a pharmaceutically acceptable salt, prodrug, metabolite, polymorph or solvate thereof, and at least a second agent as part of a specific treatment regimen intended to provide the beneficial effect from the co-action of these therapeutic agents. The beneficial effect of the combination includes, but is not limited to, a cooperative, e.g., synergistic, effect and/or a pharmacokinetic or pharmacodynamic co-action, or any combination thereof, resulting from the combination of therapeutic agents. Administration of these therapeutic agents in combination typically is carried out over a defined time period (usually minutes, hours, days or weeks depending upon the combination selected). “Combination therapy”

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PCT/EP2015/069808 may be, but generally is not, intended to encompass the administration of two or more of these therapeutic agents as part of separate monotherapy regimens that incidentally and arbitrarily result in the combinations of the present disclosure.

“Combination therapy” is intended to embrace administration of these therapeutic agents in a sequential manner, wherein each therapeutic agent is administered at a different time and in any order, or in alternation and in any order, as well as administration of these therapeutic agents, or at least two of the therapeutic agents, in a substantially simultaneous manner. Substantially simultaneous administration can be accomplished, for example, by administering to the subject a single capsule having a fixed ratio of each therapeutic agent or in multiple, single capsules for each of the therapeutic agents. Sequential or substantially simultaneous administration of each therapeutic agent can be effected by any appropriate route including, but not limited to, oral routes, intravenous routes, intramuscular routes, and direct absorption through mucous membrane tissues. The therapeutic agents can be administered by the same route or by different routes. For example, a first therapeutic agent of the combination selected may be administered by intravenous injection while the other therapeutic agents of the combination may be administered orally. Alternatively, for example, all therapeutic agents may be administered orally or all therapeutic agents may be administered by intravenous injection. The sequence in which the therapeutic agents are administered is not narrowly critical.

All percentages and ratios used herein, unless otherwise indicated, are by weight. Other features and advantages of the present disclosure will become apparent from the different examples. The provided examples illustrate different components and methodology useful in practicing the present disclosure. Generally speaking, the disclosure extends to any novel one, or any novel combination, of the features disclosed in this specification (including the accompanying claims and drawings). The examples do not limit the claimed disclosure. Thus, features, integers, characteristics, compounds or chemical moieties described in conjunction with a particular aspect, embodiment or example of the disclosure are to be understood to be applicable to any other aspect, embodiment or example described herein, unless incompatible therewith. Based on the present disclosure the skilled artisan can identify and employ other components and methodology useful for practicing the present disclosure. Moreover, unless stated otherwise, any feature disclosed herein may be replaced by an alternative feature serving the same or a similar purpose.

The Disclosure will now be described by way of example only with reference to the Examples below:

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EXEMPLIFICATION

I. COMPOUND PREPARATION

General Methods and Materials

All chemicals were purchased from Sigma-Aldrich, Alfa Aesar. 1H NMR spectra were recorded at 200 and 400 MHz and 13C NMR spectra were recorded at 100.6 and 50.3 MHz by using deuterated solvents indicated below. TLC were performed on aluminium backed silica plates (silica gel 60 F254). All the reactions were performed under nitrogen atmosphere using distilled solvents. All tested compounds were found to have > 95% purity determined by HPLC analysis. HPLC-grade water was obtained from a tandem MilliRo/Milli-Q apparatus. The analytical HPLC measurements were made on a Shimadzu LC20AProminence equipped with a CBM-20A communication bus module, two LC-20AD dual piston pumps, a SPD-M20A photodiode array detector and a Rheodyne 77251 injector with a 20 pL stainless steel loop.

Scheme 1: Preparation of Intermediate 1.4

Example 1: Preparation of Intermediate 1.4

To a stirred solution of compound 1.1 (0.52 ml, 4.9 mmol), 1.2 (372 mg, 4.9 mmol) and 1.3 (0.5 mL, 0.83 mL, 4.9 mmol) in ethanol (25 mL) was added K2CO3 (812 mg, 5.88 mmol). Stirring was continued at reflux overnight. The pale yellow solid was collected after cooling, taken up with boiling water and filtered again. The aqueous phase was acidified to pH 5 with AcOH (15 drops), the precipitate was filtered and dried under vacuum. The title compound 1.4 was obtained as pale yellow solid (500 g, 2.18 mmol). Yield 44%.

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Scheme 2: Preparation of Intermediate 2.2

NC CO2Et +

H2N nh2

2.1

Reflux

K2CO3, EtOH,

2.2

1.1 1.2

Example 2: Preparation of Intermediate 2.2

To a stirred solution of compound 1.1 (0.96 g, 8.8 mmol), 1.2 (672 mg, 8.8 mmol) and 2.1 (1g, 0.83 mL) in ethanol (55 mL) was added K2CO3 (1.57 g, 11.44 mmol). Stirring was continued at reflux overnight. The yellowish solid was collected after cooling, taken up with hot water and filtered again. The aqueous phase was acidified to pH 1, the precipitate was filtered and dried under vacuum. The title compound 2.2 was obtained as yellowish solid (1 g, 4.25 mmol). Yield 49%.1HNMR (200 MHz, DMSO) δ 7.22 (m, 1H), 7.68 (m, 1H), 7.85 (d, J = 4.8 Hz, 1H), 8.05 (s, 1H).

Scheme 3: Preparation of Intermediate 3.2

O

1.1 1.2 3.1 3.2

Example 3: Preparation of Intermediate 3.2

To a stirred solution of compound 1.1(0.96 mL, 8.8 mmol), 1.2 (672 mg, 8.8 mmol) and 3.1 (1g, 1.29 mL) in ethanol (55 mL) was added K2CO3 (1.57 g, 11.44 mmol). Stirring was continued at reflux overnight. The yellowish solid was collected after cooling, taken up with hot water and filtered again. The aqueous phase was acidified to pH 1, the precipitate was filtered and dried under vacuum. The title compound 3.2 was obtained as yellowish solid (1 g, 4.25 mmol). Yield 49%.

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Scheme 4: Preparation of Intermediate 4.2

1.1 1.2 4.1 4.2

Example 4: Preparation of Intermediate 4.2

To a stirred solution of compound 1.1 (1.42 mL, 13.37 mmol), 1.2 (1.01 g, 13.3 mmol) and 4.1 (1.62 mL, 13.3 mL) in ethanol (50 mL) was added piperidine (2.64 mL, 26.7 mmol). Stirring was continued at reflux overnight. The solid was collected after cooling, taken up with hot water and filtered again. The aqueous phase was acidified to pHl and extracted with EtOAc (3 x 25 mL). The organic phase was washed with brine and dried over Na2SO4.The crude of reaction was purified by flash chromatography ICHCL/McOH as gradient, from 0 to 2% for product), affording the title compound 4.2 (930 mg, 3.95 mmol) as white solid. Yield 30%.

Scheme 5: Preparation of Intermediate 5.2

1.1 1.2 5.1 5.2

Example 5: Preparation of Intermediate 5.2

To a stirred solution of compound 1.1 (0.49 mL, 4.67 mmol), 1.2 (355 mg, 4.67 mmol) and 5.1 (0.44 mL, 4.67 mmol) in ethanol (25 mL) was added K2CO3 (773 mg, 5.6 mmol). Stirring was continued at reflux overnight. The white solid was collected after cooling, dried under vacuum and used for the next step without further purification. The title compound 5.2 was obtained as white solid (300 mg, 1.3 mmol). Yield 29%?H NMR (400 MHz, DMSO) δ 7.64 (d, J = 4.7 Hz, 2H), 8.78 (d, J = 4.7 Hz, 2H), 12.98 (s, 1H).

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Scheme 6: Preparation of Intermediate 6.2

NC״CO2Et

X ^OEt

O

X h2n nh2

EtO־Na+, EtOH reflux

6.1 1.2 6.2

Example 6: Preparation of Intermediate 6.2

To a stirred solution NaOEt (1.02 mL, 2.73 mmol) in EtOH abs (20 mL) was added compound 6.1 (500 mg, 2.73 mmol) and 1.2 (207 mg, 2.73 mmol). Stirring was continued at reflux 4 h. The volatiles were removed under vacuum. The crude of reaction was taken up with water and acidified with AcOH. The precipitate was collected dissolved in water, washed with a mixture of CHC13 and MeOH. The aqueous phase was extracted with EtOAc (3 x 20 mL). The collected organic phase was washed with brine, dried over Na2SO4. The title compound 6.2 was obtained as white solid (250 mg, 1.49 mmol). Yield 55%.

Scheme 7: Preparation of Intermediate 7.3

Example 7a: Preparation of Intermediate 7.2

To a stirred solution of compound 1.1 (0.14 mL, 1.3 mmol) and 7.1 (150 mg, 1.3 mmol) in EtOH (5 mL) was added piperidine (1 drop). Stirring was continued at room temperature overnight. The solvent was removed under vacuum. The crude of reaction was purified by flash chromatography affording the title compound 7.2 (160 mg, 0.77 mmol) as yellowish solid. Yield 58%.

Example 7b: Preparation of Intermediate 7.3

To a stirred suspension of compound 7.2 (150 mg, 0.72 mmol) and compound 1.2 (55 mg, 0.72 mmol) in EtOH (5 mL) was added K2CO3 (99 mg, 0.72 mmol). Stirring was continued at reflux overnight. The white precipitate was collected and used as well for the

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PCT/EP2015/069808 next step without further purification. The title compound 7.3 (150 mg, 0.48 mmol) was obtained as yellowish solid as z/z-potassium salt. Yield 67%.

Scheme 8: Preparation of Intermediate 8.5

NaHCO3 NH2OH*HCI,

H2O, EtOH, 80 °C;

NH

CIOCOOEt, Me2CO,

NaOH, H2O

8.1 8.2 8.3

8.4

8.5

Example 8a: Preparation of Intermediate 8.2

To a stirred solution of NH2OH*HC1 and NaHCO3 in water (7 mL) was gradually added a solution of m-tolunitrile (8.1) (2 mL, 17.0 mmol) in EtOH (13.3 mL). Stirring was continued at 80°C for 4 h. The volatiles were removed under vacuum. The crude of reaction was taken up with water, extracted with EtOAc (3 x 25 mL). The organic phase were collected, washed with brine and dried over Na2SO4 affording the title compound 8.2 (1.5 g, 9 mmol) as white solid. Yield 59%.

Example 8b: Preparation of Intermediate 8.3

To a solution of compound 8.2 (1g, 6 mmol) in dry acetone (5 mL), was added dropwise at 0 °C EtOCOCl (0.63 mL, 6.6 mmol). Stirring was continued at this temperature for Ih. Then a 5% NaOH solution was added to the mixture. Stirring was continued for additional Ih. The solvent ws removed under vacuum. The crude of reaction was poured in water, extracted with EtOAc (3 x 50 mL). The collected organic phase was washed with brine, dried over Na2SO4. The title compound 8.3 (600 mg, 2.7 mmol) was obtained as white solid. Yield 45%.

Example 8c: Preparation o f Intermediate 8.4

To a solution of compound 8.3 (300 mg, 1.35 mmol) in EtOH abs (5 mL) was added sodium (50 mg) portion wise. Stirring was continued at room temperature for additional 4h. The reaction was quenched by the addition of MeOH. The solvent was removed under reduced pressure and the crude was purified by flash chromatography. The title compound 8.4 (150 mg, 0.85 mmol) was obtained as white solid. Yield 63%.

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Example 8d: Preparation of Intermediate 8.5

To a suspension of compound 8.4 (326 mg, 1.85 mmol) in CC14 (10 mL) was added AIBN (60.7 mg, 0.37 mmol) and NBS (493 mg, 2.77 mmol). Stirring was continued at reflux overnight. The solvent was removed under reduced pressure. The reaction was taken up with water, extracted with EtOAc (3 x 20 mL) washed with brine and dried over Na2SO4.The crude was purified by flash chromatography, eluting with Petroleum ether (Pet. Ether) /EtOAc (30% for product) affording the title compound 8.5 (280 mg, 1.09 mmol) was obtained as white solid. Yield 59%.

Scheme 9: Preparation of Intermediate 9.2

NBS, AIBN, Br

CCI41 reflux;

9.1 9.2

Example 9: Preparation of Intermediate 9.2

To a suspension of compound 9.1 (750 mg, 5 mmol) in CC14 (15 mL) was added AIBN (41 mg, 0.25 mmol) and NBS (933.7 mg, 5.24 mmol). Stirring was continued at reflux overnight. The solvent was removed under reduced pressure. The reaction was taken up with water, extracted with EtOAc (3 x 20 mL) washed with brine and dried over Na2SO4.The crude was purified by flash chromatography, eluting with CH2C12/MeOH (3% for product) affording the title compound 9.2 (800 mg, 3.49 mmol) as white solid. Yield 70%.

Scheme 10: Preparation of Intermediate 10.

N'N n-n

Et3NH+CI-, NaN3N NBS, AIBN, '

h ץ ץ ־·---- ״it ״------toluene, 110°C CH3CN, reflux;

10.2 10.3

Example 10a: Preparation of Intermediate 10.2

A mixture of compound 10.1 (1.02 mL, 8.54 mmol), NaN3 (832 mg, 12.8 mmol) and Et3N*HCl (1.76 g, 12.8 mmol) was heated at reflux 4 h. The solvent was removed under vacuum. The crude was poured in water, acidified to pH 1 with 3N HC1 and extracted with EtOAc (3 x 20 mL). The organic phase was washed with brine, dried over Na2SO4 and

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Example 10b: Preparation of Intermediate 10.3

Scheme 27: Preparation of Compound 17

K2CO3 ch3cn reflux

2.2 10.3 17

Example 27: Preparation of Compound 17

To a stirred suspension of intermediate 2.2 (160 mg, 0.66 mmol) and DIPEA (0.09 mL, 0.55 mmol) in DMSO (3 mL) was added intermediate 10.3 (171 mg, 0.55 mmol). Stirring was continued overnight at room temperature. The crude was poured in water, washed with EtOAc then acidified to pH 3 and extracted with EtOAc (3 x 50 mL). The title compound 17 has been obtained (90 mg, 0.22 mmol) as pure orange solid after flash chromatography purification eluting with CH2C12/MeOH (5% for product) and prior titration with a mixture of Et2O/Acetone. Yield 23%. 1H NMR (400 MHz, DMSO) δ 4.59 (s, 2H), 7.29 (t, 7= 4.6 Hz, 1H), 7.54 (t, 7= 7.6 Hz, 1H), 7.68 (t, 7= 7.8 Hz, 1H), 7.91 (d, 7 = 7.7 Hz, 1H), 7.96 (d, 7 = 4.9 Hz, 1H), 8.16 (s, 1H), 8.22 (d, 7= 3.8 Hz, 1H); 13C NMR (100 MHz, DMSO) δ 33.9, 88.9, 117.6, 125.0, 126.2, 127.9, 129.6, 129.9, 131.2, 131.9, 134.4, 139.3, 140.4, 155.8, 159.1, 163.8, 167.0. HPLC 96.2%.

Scheme 28: Preparation of Compound 23

2.2

K2CO3, Acetone

11.2 23

Example 28: Preparation of Compound 23

To a stirred suspension of intermediate 2.2 (150 mg, 0.63 mmol) and K2CO3 (96.6 mg, 0.70 mmol) in acetone (10 mL) was added intermediate 11.2 (173 mg, 0.72 mmol). Stirring was continued overnight at room temperature. The volatiles were removed under vacuo. The crude was taken up with water, acidified to pH 3 and extracted with EtOAc (3 x 50 mL). The title compound 23 has been obtained (150 mg, 0.39 mmol) as pure orange solid

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PCT/EP2015/069808 after flash chromatography purification eluting with CH2C12/M6OH (5% for product) and prior titration with hot EtOAc. Yield 62%. 1H NMR (400 MHz, DMSO) δ 3.92 (s, 2H), 6.72 (t, J = ר.ר Hz, 1H), 6.79 (d, J = 7.6 Hz, 1H), 6.88 (t, J = 7.3 Hz, 1H), 7.20 (t, J = 4.26 Hz, 1H), 7.77 (d, J = 4.7 Hz, 1H); 7.92 (d, 7= 7.8 Hz, 1H), 8.0 (d, 7= 3.5 Hz, 1H); 9.59 (s, 1H), 9.80 (s, 1H); 13C NMR (100 MHz, DMSO) δ 35.12, 86.2, 115.5, 119.2, 120.1, 121.0, 124.4, 127.0, 128.7, 129.2, 131.4, 141.8, 147.3, 159.2, 167.9, 169.1, 170.5. HPLC 97.7%.

If. BIOLOGICAL ACTIVITY

Example 29: Determination of ACMSD1 inhibition

The activity of compounds 1-19 and 21-23 as inhibitors of ACMSD 1 was determined by measuring the conversion of 3OH-Anthranilic Acid into product (i.e., ACMS) in a spectrophotometrical in vitro assay.

The pre-assay mixture consisting of 3-hydroxyanthranilic acid (3OH-HA), 3hydroxyanthranilic acid, 3,4-diOxygenase (HAO), and a dialyzed crude extract of E. coli BL21 (DE3) cells expressing the recombinant enzyme, was incubated at 25 °C with monitoring of the increase in absorbance at 360 nm due to the formation of ACMS from 3OH-HA. After the reaction was completed within ~ 2 mins, an aliquot of ACMSD 1 solution (prepared and purified from Pichia Pastoris overexpressing the recombinant enzyme) was added, and the decrease in absorbance at 360 nm was followed at 15 second intervals. The effect of ACMS concentration on the enzyme activity was investigated by varying 3OH-HA concentration from 2 to 20 μΜ. Kinetic parameters were calculated from the initial velocity data by using the Lineweaver-Burk plot.

The rate of the decrease in absorbance caused by ACMSD 1 was calculated by subtracting that of the control reaction mixture without ACMSD from that described above. One unit of ACMSD activity was indicated as the amount of enzyme that converts 1 mmol of ACMS per minute at 25 °C. The absence or a reduction of ACMSD 1 activity (e.g., by using ACMSD inhibitors) results in a slow ACMS-spontaneous degradation (i.e., cyclization to form quinolic acid).

The enzymatic activity was determined at a HAA concentration of 10μΜ in the presence of the compounds in Table 1 below. The compounds were tested at the concentration of about 5 μΜ and 10μΜ and the IC50 was calculated for compounds showing inhibitory activity higher than 50%. The results are shown in Table 1.

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TABLE 1:

<td> Compound No.</td><td> Structure</td><td> Activity hACMSD IC50</td>

<td> 1</td><td> 0 ncAnh</td><td> 0.050</td>

<td> Compound No.</td><td> Structure</td><td> Activity hACMSD IC50</td>

Example 30: Determination of ACMSD-1 modulation in HEK293T cells

HEK293T cells (ATCC) were seeded in six-well plates and transfected using Fugene HD to express transiently ACMSD. 24 hrs post transfection, the cells were stimulated for 48 hrs to 72 hrs with different concentrations of Compound 1 and then lysed to measure the ACMSD activity, by measuring the conversion of 3 OH-Anthranilic Acid into product (i.e. aamino-beta-carboxymuconate-8-semialdehyde, ACMS) in a spectrophotometrical in vitro assay. The amount of the whole protein content in cell lysates was detected by Bradford analysis. This value was used to get the specificity activity of the enzyme normalized in all samples (mU/ml or ΔΕ/At/mg of total protein).

ACMSD-1 enzyme is known to be expressed in liver, kidney and brain; available cell lines for these cell types were therefore tested to determine the expression levels of ACMSD. We determined that ACMSD-1 is not expressed in transformed cell lines from liver and

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PCT/EP2015/069808 kidney, such as HepG2, HEK293T, Hep3B etc. Transfection of ACMSD was performed to express the enzyme in different cellular backgrounds such as COS-7, HEK293T, and HepG2. The HEK293T cellular background proved to be the best system, with the highest protein production allowing robust measurement ACMSD 1 enzyme activity. This is probably due to the better transfection efficacy observed in HEK293T.

Having determined the optimum stimulation time and transfection protocol cells were stimulated with different concentrations of Compound 1 (about 50 nM to about 5 uM). Compound 1 inhibited ACMSD-1 activity, in a dose dependent manner, in this overexpression cell-based assay.

Example 31: Determination of NAD+ content in Human Primary Hepatocytes Treated with Compound 4

The NAD+ concentration or content was determined in human primary hepatocytes treated with Compound 4. Vehicle (NT) was used as a control.

At least three experiments were run treating primary hepatocytes with different concentrations of Compound 4 (0.5 μΜ and 5 μΜ) after 48 hrs from seeding. Compound 4 was replaced every 24 hrs, and then cells were directly harvested and lysed with ACN/H2O (ratio 5:1). LCMS/MS was used to detect and measure NAD+ concentration/content. Screening data showed that Compound 4 inhibits ACMSD-1 enzyme at concentrations as low as 0.5μΜ and 5μΜ. (FIG. 1)

Example 32: Determination of NAD+ content in Human Primary Hepatocytes Treated with Compound 1

The NAD+ concentration or content was determined in human primary hepatocytes treated with Compound 1 and MEHP, a known ACMSD inhibitor. MEHP was used as a control.

At least three experiments were run treating primary hepatocytes with different concentrations of Compound 1 (0.5 μΜ, 5 μΜ, and 50 μΜ) after 48 hrs from seeding. Compound 1 was replaced every 24 hrs, and then cells were directly harvested and lysed with ACN/H2O (ratio 5:1). LCMS/MS was used to detect and measure NAD+ concentration/content. Screening data showed that 500μΜ of MEHP inhibits 70% of purified ACMSD-1 enzyme, and that 0.5μΜ of Compound 1 has similar inhibition activity as 250μΜ of MEHP. (FIG. 3)

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<td> 8</td><td> o £j ״a'sA1 ci ^^co2h</td><td> Not Toxic</td><td> Not Toxic</td><td> No Activity</td>

<td> 4</td><td> 0 ncYnh</td><td> Not Toxic</td><td> Not Toxic</td><td> No Activity</td>

<td> 19</td><td> ץ N'Q 1 Λ (ד n ΑΓ1 & V-s YY</td><td> Not Toxic</td><td> Not Toxic</td><td> No Activity</td>

<td> 5</td><td> ץ ך! nh / N S η Vs</td><td> Not Toxic</td><td> Not Toxic</td><td> No Activity</td>

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<td rowspan="2"> Compound</td><td> Compound Structure</td><td colspan="2"> Cytotoxicity</td><td rowspan="2"> hERG</td>

<td> 1</td><td> 0 1״A</td><td> Not Toxic</td><td> Not Toxic</td><td> No Activity</td>

<td> 11</td><td> 0</td><td> Not Toxic</td><td> Not Toxic</td><td> No Activity</td>

Example 38: C.elegans experiments - ACMSD1 silencing, lifespan assays, mobility assessment and GFP fluorescence quantification

C. elegans (Caenorhabditis elegans) strains were provided by the Caenorhabditis Genetics Center (University of Minnesota). Worms were maintained on Nematode Growth Medium (NGM) agar plates seeded with E.coli OP50 bacteria at 20°C, unless stated otherwise. The strains used for the experiments were the following: Bristol N2, NL2099 (rrf3(pkl426)II), KN259 (huIs33[sod-3::GFP+pRF4(rol-6(sul006))]).

Bacterial feeding RNAi experiments were carried out as follows: worms were grown on NGM agar plates containing Carbenicillin and IPTG at final concentrations of 25 pg/ml and ImM respectively and seeded with bacterial cultures taken from Ahringer library. Clones used were acmsd-1 (Y71D11A.3), sir-2.1 (R11A8.4), and daf-16 (R13H8.1). Clones were purchased from GeneService and their identity was confirmed by sequencing. For the double RNAi experiments bacterial cultures were mixed before seeding on NGM plates. The control RNAi in this kind of experiments was 50% diluted with control empty vector RNAi bacteria.

The nematode Caenorhabditis elegans was used as a model system to confirm the activation of the oxidative stress defence that we have observed in cells at the level of an intact organism. The effects of acmsd-1 RNAi were assessed in C. elegans by RT-qPCR.

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The total RNA was extracted from cells using TRIzol (Invitrogen) according to the manufacturer’s instructions. RNA was treated with DNase, and 2 pg of RNA was used for reverse transcription (RT). 50X diluted cDNA was used for RT-quantitative PCR (RTqPCR) reactions. The RT-qPCR reactions were performed using the Light-Cycler system (Roche Applied Science) and a qPCR Supermix (QIAGEN) with the indicated primers. The average of at least three technical repeats was used for each biological data point.

C. elegans lifespan assays were carried at 20 °C as follows. Animals were exposed to NAC (N-acetyl cysteine) at a final concentration of 5 mM from a 0.5 M aqueous stock from the young adult stage. Sodium pyruvate was added at a final concentration of 2.5 mM to NGM plates containing carbenicillin (100 1g mL)l) and seeded with UV-killed OP50. After 5 days of RNAi treatment, worms were transferred to plates containing paraquat and seeded with acmsd-1 RNAi bacteria. Control animals were grown during the first 5 days of adulthood on RNAi bacteria containing the empty vector and then transferred to plates containing paraquat and seeded with acmsd-1 RNAi bacteria. Survival analyses were performed using the Kaplan-Meier method, and the significance of differences between survival curves was calculated using the log rank test. The statistical software used was XLSTAT 2007 (XLSTAT, Brooklyn, NY, USA), and all P-values < 0.05 were considered significant. 100 worms were used per condition and scored every 2 days. The reasons for censoring were the «exploded vulva» phenotype or worms that crawled off the plate. Where indicated, paraquat was added on top of the agar plates at the indicated concentration. Once the paraquat solution was completely dried, L4 worms were transferred to these agar plates and monitored for 5-6 days every day. By day 6 all the paraquat tests were stopped because a small percentage in worm population could start to die naturally and rather than dying due to the paraquat effects.

The movement of worms was recorded for 45 seconds at days 1,3, and 5 of adulthood using a Nikon DS-L2 / DS-Fil camera and controller setup, attached to both a computer and a standard bright field microscope. For each condition five plates of worms, with 10 worms per plate were used. The movement of worms during aging was calculated by taking an integral of the speed value which was assessed by following the worm centroids with a modified version of the freely-available for the Parallel Worm Tracker for MATLAB.

Fluorescence intensity in worm strains expressing GFP-reporter proteins was quantified using Victor X4 plate reader (Perkin Elmer). The animals were prepared in the following way: eighty worms per condition (at the corresponding ages) were picked (20

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PCT/EP2015/069808 worms per well of a black-walled 96-well plate) and placed into the M9 medium. Each experiment was repeated at least twice.

The expression level of acmsd-1 mRNA was significantly reduced confirming the efficacy of RNAi mediated gene knock-down (FIG. 7A). The worm ortholog of MnSOD, SOD-3, was induced at its mRNA level with concomitant downregulation of the acmsd-1 gene with RNAi. A significant increase at the protein level of SOD-3 was also observed at Day 3 of adulthood. (FIG. 7B) ACMSD downregulation improved worm lifespan and this improvement was SIR-2.1- and DAF-16-dependent. (FIG. 7C)

Analyses of disease onset and survival

Disease onset is defined as the last day of individual peak body weight before gradual loss occurs. The stages of disease are defined as follows: the early stage of disease is defined as the duration of time between peak body weight until loss of 10% of peak body weight. The late stage of disease is defined as the duration of time between 10% loss of peak body weight until the end stage of disease. The end stage of disease is defined as the day when an animal could no longer right itself within 30 s for three consecutive trials when placed on its side. Animals are euthanized at the end stage of disease.

Body composition measurements

Body weights are assessed weekly for at least 13 weeks. Brown adipose tissue (BAT) and gonadal white adipose tissue (WAT) are dissected and weighed at the indicated age. Total lean mass, % of WAT and BMD (bone mineral density) are determined by DEXA (PIXImus DEXA; GE).

Indirect calorimetry, food intake and activity

Animals are initially weighed and acclimated to the test cage. Volume oxygen (VO2) and volume carbon dioxide production (VCO2) are measured every 20 min using the Oxymax Comprehensive Laboratory Animal Monitoring System (CLAMS) (Columbus Instruments) and are reported as average VO2 per hour normalized to body weight (mL/h/kg). Using the CLAMS machine, activity counts by infrared beam interruptions and food intake are simultaneously measured. More specifically, food intake is measured by deducting the weight of powderized food pellets at the end of experimentation from the starting weight at the beginning of experimentation. To complement this experiment and to control for a novel environment that may affect feeding behaviour, we also perform a more ‘manual’ experiment, wherein a set weight of food pellets is placed at the same time each day into a clean home cage, which holds a mouse. The next day the weight of the remaining pellets is recorded and deducted from the starting weight. This experiment is performed for 14 days straight. The body weight of each mouse is also recorded daily. Results for each genotype are similar to that acquired from the CLAMS.

Statistical analyses.

Considering a l-β larger than 0.9 statistically powerful, we estimate appropriate group numbers from pilot studies a priori. One- or two-way Analyses of Variance (Bonferroni post-tests) or t-tests are performed.

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Example 41: Study of the Effects of Compounds of Formula CD, Formula (la). Formula (lb). Formula (II), or Formula (III) on Non-alcoholic Fatty Liver Disease (NAFLD) and Non-alcoholic Steatohepatitis (NASH) in Mice

A study is performed to determine the effects of compounds of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, on non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) in male C57BL/6I fed a high fat and high sucrose diet.

PCT/EP2015/069808

Compounds of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, are dissolved in water and added to a standard powder diet (Mucedola, Italy) at the appropriate concentration of 50 mg/Kg/day. Pellets containing the compounds of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, or the vehicles are reconstituted by hand and kept frozen at - 20°C until needed. The diet supply is changed every three days, and only the amount needed is thawed at each time and administered ad libitum for one month. Sco2KO/KI mice are maintained in a temperature- and humidity-controlled animal-care facility, with a 12 hrs light/dark cycle and free access to water and food. Animals are sacrificed by cervical dislocation.

Morphological Analysis

For histochemical analysis, tissues are frozen in liquid-nitrogen precooled isopentane. Series of 8 mm thick sections are stained for COX and SDH.

Biochemical Analysis ofMRC Complexes

Muscle quadriceps samples stored in liquid nitrogen are homogenized in 10 mM phosphate buffer (pH 7.4), and the spectrophotometric activity of cl, ell, cIII, and cIV, as well as CS, is measured as described. Note that in all panels the activity of ell is multiplied by 10 for visualization clarity.

PCT/EP2015/069808 instructions (BioVision, Milpitas, CA). In addition, the proinflammatory cytokines TNF-a, IL-lb, and IL-6 from serum or homogenates from kidney tissue are quantified by ELISA (Quantikine Kit; R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions. For measuring cytokines, kidney tissue is homogenized in phosphate buffered saline containing 0.05% Tween-20. Aliquots containing 300 mg of total protein are used. A metabolic cage is used for collecting urine to analyse the level of urinary cytokines. The sample size for each group is five.

Alternative Study of the Effects of Compounds of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III) on Kidney Disease

Alternatively, C57BL/6J WT mice are numbered and kept in acclimatization for a period of 5-7 days before initiation of the experiment. (Wei, Q., et al.. “Mouse model of ischemic acute kidney injury: technical notes and tricks” American Journal of PhysiologyRenal Physiology, 303(11), F1487-F1494) Mice are randomized into different treatment groups based on their body weight. Different groups are maintained on Harlan diet 2916. Mice are then maintained on the respective diets for 10 days prior to bilateral Ischemic kidney injury. Body weight measurement is made once at randomization and once on day 7. Food consumption is evaluated once on day 7. Blood is collected by retro-orbital puncture under mild Isoflurane anesthesia and used for analysis of basal blood urea nitrogen levels (BUN) on day 9.

Mice are anesthetized with ketamine (80 mg/kg i.p.) and/or Xylazine (10 mg/kg, i.p.) and placed on a surgical platform in a dorsal position. Both kidneys are exposed through flank incisions and renal pedicles are occluded using vascular clamps for 25 minutes. The clamp is then removed and the surgical site is sutured. 1ml of physiological saline is administered intra-peritoneally after closing the wound to prevent dehydration. The shamoperated group is subjected to similar surgical procedures, except that the occluding clamp is not applied. Animals are monitored until recovery from anesthesia and returned to their home cage. Animals are observed every day for general clinical signs and symptoms and mortality.

One day prior to termination, animals are individually housed in metabolic cages for 12h and urine is collected for estimation of urea, creatinine, sodium and potassium.

On days 12, 14, & 16 blood is collected by retro orbital puncture under mild isoflurane anesthesia and plasma is used for analysis of blood urea nitrogen levels (BUN) and serum creatinine. Animals are then euthanized by CO2 inhalation and organs are collected. One kidney is fixed in 10% neutral buffered formalin and the other is flash frozen in liquid

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PCT/EP2015/069808 nitrogen, stored at -80°C and used for the estimation of lipid peroxidation, GSH, MPO and SOD levels.

Histological analysis and neutrophil counting

Mouse kidneys are fixed in 4% formaldehyde and embedded in paraffin wax. The 5mm-thick sections are deparaffinised in xylene and rehydrated through graded concentrations of ethanol. H&E and PAS staining are performed using standard protocols. Images are collected and analysed using a light microscope (1X71, Olympus, Tokyo, Japan) with DP analyser software (DP70-BSW, Tokyo, Japan). Tubular damage in PAS-stained kidney sections is examined under a light microscope and scored based on the percentage of cortical tubular necrosis: 0 = normal, I = 1-10, 2= 11-25, 3 = 26-45, 4 = 46-75, and 5 = 76-100%. Slides are scored in a blinded manner, and results are means ± s.d. of 10 representative fields/group. Severity criterion for tubular necrosis displaying the loss of the proximal tubular brush border and cast formation are used to classify samples. The sample size for each group is 10. Neutrophil infiltration is quantitatively assessed on PAS stained tissue by a renal pathologist by counting the number of neutrophils per high-power field (x400). At least 10 fields are counted in the outer stripe of the outer medulla for each slide.

All values are represented as mean ± s.d. One-way analysis of variance is used to calculate the statistical significance of the results of all assays and P-values < 0.05 are considered statistically significant.

Example 47: Study of the Effects of Compounds of Formula (I), Formula (la). Formula (lb). Formula (II), or Formula (III) on Ischemia/Reperfusion-induced Acute Kidney Injury

CD-I (ICR) mice are purchased from Charles River Laboratory (Wilmington, MA). Mice are housed in a temperature- and humidity-controlled environment with a 12:12 hrs light-dark cycle and are allowed freely access to standard rodent chow (TekLad, Madison, WI) and tap water.

Mice are subjected to a midline back incision, and both renal pedicles are clamped for 45 min with microaneurysm clamps (00396-01; Fine Science Tools, Foster City, CA). After

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PCT/EP2015/069808 removal of the clamp, the kidneys are inspected for the restoration of blood flow. The animals are allowed to recover, and they are sacrificed 48 hrs after reperfusion. Mice are treated with 100 mg/kg of a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, by oral gavage once per day. CD-I mice are divided into four groups: (1) young mice with sham injury (n = 4) (6--7 weeks old); (2) young mice with I/R injury (n = 8); (3) adult mice with sham injury (n = 4) (20-24 weeks old); and (4) adult mice with PR injury (n = 11). An additional 27 adult mice (20-24 weeks old) are randomized into two groups: 13 mice received a compound of Formula (I), Formula (la), Formula (lb), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, and the other 14 mice received the vehicle as a control.

The serum creatinine level is measured using the QuantiChrom Creatinine Assay Kit (DICT-500, BioAssay Systems, Hayward, CA). BUN measurements are recorded using the Infinity Urea (Nitrogen) Liquid Stable Reagent (TR12421; ThermoTrace, Victoria, AU).

Evaluation of renal tissue

Kidneys are fixed in 4% paraformaldehyde, embedded in paraffin, and stained with hematoxylin and eosin (4 mm thick). Tubular injury is scored on a scale of 0-4 on the basis of the percentage of tubules with necrosis, dilatation, or cell swelling: 0, less than 5%; 1, 525%; 2, 25-50%; 3, 50-75%; and 4, over 75%. All high-power fields (x 400) in the cortex and outer medulla are evaluated by a pathologist in a blinded manner.

All values are expressed as mean ± s.e. Statistical analysis is carried out using GraphPad Prism 4.00 (San Diego, CA) with unpaired Student’s t testing for two sets of data and an analysis of variance with a Bonferroni post-test for multiple groups. P < 0.05 was considered significant.

Example 48: Determination of the Effects of Compounds 1 and 17 on FoxOl Phosphorylation levels

AML-12 cells were treated with different concentrations of Compound 1 or Compound 17 for 24 hours. Cells were then lysed in lysis buffer (50 mM Tris, 150 mM KC1, EDTA IrnM, NP40 1%) containing protease and phosphatase inhibitors, and analyzed by SDS-PAGE/westem blot. Blocking and antibody incubations were done in 5% milk. Each protein present was detected with its specific antibody. Tubulin antibody was obtained from Sigma Inc, FoxOl and phopho-FoxOl (Ser256) antibodies were obtained from Cell Signaling. Antibody detection reactions were developed by enhanced chemiluminescence (Advansta, CA, USA) using x-ray films.

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FoxOl phosphorylation at Ser256 results in its nuclear export and in inhibition of its transcription factor activity. A decrease in FoxOl phosphorylation at Ser256 with increasing dose of Compound 1 and 17 was observed (FIG. 6), indicating an increase in nucleartranslocated FoxOl and therefore increased FoxOl transcriptional activity.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the specification, the singular forms also include the plural unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice of testing the present disclosure, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are hereby expressly incorporated by reference. The references cited herein are not admitted to be prior art of the claimed disclosure. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting.

Example 49: Synthesis of Exemplified Compounds

2)H2, 10% Pd/C, EtOH, THF, rt, 2 h

1) diethyl (cyanomethyl) phosphonate, K2CO3, THF, H2O, 60 C, 1 h;

MeO2C

2a 2b 2c

See, Hirose M, et al., “Design and synthesis of novel DFG-out RAF/vascular endothelial growth factor receptor 2 (VEGFR2) inhibitors: 3. Evaluation of 5-amino-linked thiazolo[5,4-d]pyrimidine and thiazolo[5,4-b]pyridine derivatives.” Bioorg. Med. Chem. 2012, 15;20(7 8):5600-15.

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Benzoylchloroformate,

NaHCO3 H2O, Dioxane

78%

CH2N2 MeOH Wolff rearr.

7b 7c 7d

2M, NaOH, MeOH

See, Clift MD, Silverman RB., “Synthesis and evaluation of novel aromatic substrates and competitive inhibitors of GABA aminotransferase,” Bioorg. Med. Chem. Lett,. 2008,

15;18(10):3122-5.

base, DMF

X = Cl, Br, SMe, SO2Me

2M, NaOH, MeOH

A. M. El-Reedy, A. O. Ayyad and A. S. Ali, “Azolopyrimidines and pyrimidoquinazolines from 4-chloropyrimidines,” J. Het. Chem. 1989, 26, 313-16.

11c

; DMF, CO2CI2 I DME, 40 °C ' ♦

base, DMF

2M, NaOH, MeOH

11f Commercial

See, Iwahashi M, et al., “Design and synthesis of new prostaglandin D2 receptor antagonists,” Rzoorg. Med. Chem. 2011, 19(28):5361-71.

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2M, NaOH, MeOH

13d

NH3-THF, THF, rt, 34h, rt

13e

See, U.S. 2008/004,302(Al); and U.S. 8,716,470 (B2).

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments and methods described herein. Such equivalents are intended to be encompassed by the scope of the present disclosure.

Contents263

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Priority claims
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201462043853	United States of America	P	2014-08-29
201462043853	United States of America	P	2014-08-29
62043853	United States of America	–	2014-08-29
2015069808	European Patent Office (EPO)	W	2015-08-28
2015069808	European Patent Office (EPO)	W	2015-08-28
62043853	–	–	–
PCTEP2015069808	–	–	–
US201462043853P	–	–	–
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IL250730D0	Israel	D0	2017-04-30
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US9708272B2	United States of America	B2	2017-07-18
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Abstract

Term

RELATED APPLICATIONS

FIELD OF THE DISCLOSURE

BACKGROUND OF THE DISCLOSURE

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SUMMARY OF THE DISCLOSURE

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BRIEF DESCRIPTION OF THE DRAWINGS

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DETAILED DESCRIPTION OF THE DISCLOSURE

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