TREATING LONG QT SYNDROME

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR

DEVELOPMENT

The present invention was made with government support under Grant No.

1R01HL132905 awarded by the National Institutes of Health. The United States

Government has certain rights in the invention.

CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application Serial No.

62/800,991, filed February 4, 2019, the disclosure of which is incorporated herein by reference in its entirety.

TECHNICAL FIELD

This disclosure relates to compounds useful for treating and preventing disorders associated with long QT syndrome such as cardiac arrhythmia, ventricular arrhythmia, hypertrophic cardiomyopathy, and congestive heart failure. Also provided herein are methods and materials for using such compounds to shorten myocardial repolarization time in a patient.

BACKGROUND

Long QT syndrome (LQTS) affects one in 3,000 live births and is responsible for approximately 4,000 deaths in the United States per year. The syndrome is defined clinically as an increased interval between the onset of the Q wave and the end of the T wave on the electrocardiogram, which represents a prolongation of the myocardial repolarization time. LQTS can either be congenital or acquired as a result of medication or metabolic disturbance. Many congenital forms of LQTS have been described, but the majority of cases are the result of mutations in one of three cardiac ion channel genes: KCNQ1, KCNH2, or SCN5a.

Mutations in KCNQ1 or SCN5a lead to LQTS 1 and 3, while defects in KCNH2 (also known as the human ether-a-go-go related gene hERG ) lead to LQTS 2.

SUMMARY

The present disclosure provides a compound of Formula (I):

or a pharmaceutically acceptable salt thereof,

wherein:

each R ¹ is independently selected from the group consisting of Ci- ₆ alkyl, Ci- ₆ alkoxy, and Ci- ₆ haloalkyl;

L is a bond or selected from the group consisting of C(O), C(0)NH, C(0)NCH ₃ , NHCO, NCH C(0), SO2NH, and NHS0 ₂ ;

A is a bond or a 5-membered heteroaryl; wherein the 5-membered heteroaryl has at least one ring-forming carbon atom and 1, 2, or 3 ring-forming N atoms;

Cy is selected from the group consisting of C6-10 aryl, 5-10 membered heteroaryl, and 4-14 membered heterocycloalkyl; wherein each 5-10 membered heteroaryl and 4-14 membered heterocycloalkyl has at least one ring-forming carbon atom and 1, 2, 3, or 4 ring forming N or S atoms; wherein the one or more ring-forming carbon atoms of the 5-10 membered heteroaryl and 4-14 membered heterocycloalkyl are optionally substituted by oxo to form a carbonyl group; and wherein the C6-10 aryl, 5-10 membered heteroaryl, and 4-14 membered heterocycloalkyl are each substituted with n R ⁶ ;

each R ⁶ is independently selected from the group consisting of Ci- ₆ alkyl, Ci- ₆ alkoxy, Ci- ₆ haloalkyl, and C(0)NH ₂ ;

m is 0, 1, 2, 3, 4 or 5; and

n is 0, 1, 2, 3, 4, or 5;

provided that when L is C(0)NH or NHC(O) and A is a bond, Cy is not unsubstituted or substituted phenyl.

In some embodiments, each R ¹ is independently selected from the group consisting of Ci- ₆ alkyl, Ci- ₆ alkoxy, and Ci- ₆ haloalkyl;

L is a bond or selected from the group consisting of C(O), C(0)NCH ₃ , NCH ₃ C(0), SO2NH, and NHSO2;

A is a bond or a 5-membered heteroaryl; wherein the 5-membered heteroaryl has at least one ring-forming carbon atom and 1, 2, or 3 ring-forming N atoms; Cy is selected from the group consisting of C6-10 aryl, 5-10 membered heteroaryl, and 4-14 membered heterocycloalkyl; wherein each 5-10 membered heteroaryl and 4-14 membered heterocycloalkyl has at least one ring-forming carbon atom and 1, 2, 3, or 4 ring forming N or S atoms; wherein the one or more ring-forming carbon atoms of the 5-10 membered heteroaryl and 4-14 membered heterocycloalkyl are optionally substituted by oxo to form a carbonyl group; and wherein the C6-10 aryl, 5-10 membered heteroaryl, and 4-14 membered heterocycloalkyl are each substituted with n R ⁶ ;

each R ⁶ is independently selected from the group consisting of Ci- ₆ alkyl, Ci- ₆ alkoxy, Ci- ₆ haloalkyl, and C(0)NH2;

m is 0, 1, 2, 3, 4 or 5; and

n is 0, 1, 2, 3, 4, or 5.

In some embodiments, Cy is selected from the group consisting of phenyl, benzoimidazolyl, tetrahydroisoquinolinyl, pyridyl, phenothiazinyl, dihydroisoquinolinonyl and isoindolinonyl; wherein the phenyl, benzoimidazolyl, tetrahydroisoquinolinyl, pyridyl, phenothiazinyl, dihydroisoquinolinonyl and isoindolinonyl are each substituted with n R ⁶ . In some embodiments, Cy is phenyl substituted with 1 or 2 R ⁶ .

In some embodiments, provided herein is a compound of Formula la, Formula lb, Formula Ic, or Formula Id:

or a pharmaceutically acceptable salt thereof,

wherein Xi and X ₂ are each N or CR ^X , provided both Xi and X ₂ are not both N;

Y is CH ₂ , CH ₂ -CH ₂ , or CH-N; and R ^x is hydrogen or R ⁶ .

In some embodiments, provided herein is a compound of Formula Ial, Formula Ia2, or Formula Ia3:

Ia3,

or a pharmaceutically acceptable salt thereof,

wherein one of Xi and X2 is N and the other of Xi and X2 is CR ^X , and R ^x is hydrogen or R ⁶ .

In some embodiments, provided herein is a Formula lb 1, Formula Ib2, or Formula Ib3:

or a pharmaceutically acceptable salt thereof.

In some embodiments, provided herein is a Formula Idl, Formula Id2, or Formula

Id3:

Id3,

or a pharmaceutically acceptable salt thereof.

In some embodiments, R ¹ is Ci- ₆ alkyl. In some embodiments, R ¹ is Ci- ₆ alkoxy. In some embodiments, R ¹ is Ci. ₆ haloalkyl. In some embodiments, each R ¹ is independently methyl, methoxy, or trifluorom ethyl. In some embodiments, each R ¹ is independently methoxy or trifluorom ethyl. In some embodiments, R ¹ is methoxy.

In some embodiments, each R ⁶ is independently Ci- ₆ alkyl, Ci- ₆ alkoxy, or Ci- ₆ haloalkyl. In some embodiments, each R ⁶ is independently Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some embodiments, R ⁶ is Ci- ₆ alkyl. In some embodiments, R ⁶ is Ci- ₆ haloalkyl. In some embodiments, each R ⁶ is independently methyl, trifluoromethyl, or C(0)NH _2.

In some embodiments, n is 0, 1 or 2. In some embodiments, n is 1.

In some embodiments, the compound is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof. In some embodiments, the present disclosure also includes a pharmaceutical composition comprising a compound provided herein and one or more pharmaceutically acceptable excipients.

Provided herein is a method of treating or preventing cardiac arrhyth ia, ventricular arrhythmia, hypertrophic cardiomyopathy, or congestive heart failure, comprising administering to a patient in need thereof a therapeutically effective amount of a compound provided herein, or a pharmaceutically acceptable salt thereof.

This disclosure also provides a method of shortening myocardial repolarization time comprising administering to a patient in need thereof a therapeutically effective amount of a compound of provided herein, or a pharmaceutically acceptable salt thereof.

In some embodiments, the patient has long QT syndrome. In some embodiments, the patient has long QT syndrome type 2. In some embodiments, the long QT syndrome is drug induced long QT syndrome.

In some embodiments, the administration of the compound is acute. In some embodiments, the administration of the compound is chronic.

Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound (2MMB, 50 uM) treated.

FIG. 2 shows a duplicate LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound (2MMB, 50 uM) treated.

FIG. 3 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound (50 uM) treated.

FIG. 4 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound (5 uM) treated. FIG. 5 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 1 (50 uM) treated.

FIG. 6 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 2 (50 uM) treated.

FIG. 7 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 3 (50 uM) treated.

FIG. 8 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 4 (50 uM) treated.

FIG. 9 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 5 (50 uM) treated.

FIG. 10 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 6 (50 uM) treated.

FIG. 11 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 7 (50 uM) treated.

FIG. 12 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 8 (50 uM) treated.

FIG. 13 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 9 (50 uM) treated.

FIG. 14 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 10 (50 uM) treated.

FIG. 15 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 11 (50 uM) treated.

FIG. 16 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 12 (50 uM) treated.

FIG. 17 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 13 (50 uM) treated.

FIG. 18 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 14 (50 uM) treated.

FIG. 19 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 15 (50 uM) treated.

FIG. 20 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 16 (50 uM) treated. FIG. 21 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 18 (5 uM) treated.

FIG. 22 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 19 (50 uM) treated.

FIG. 23 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 19 (50 uM) treated.

FIG. 24 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 19 (5 uM) treated.

FIG. 25 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 20 (50 uM) treated.

FIG. 26 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 20 (5 uM) treated.

FIG. 27 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 21 (50 uM) treated.

FIG. 28 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 21 (5 uM) treated.

FIG. 29 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 22 (50 uM) treated.

FIG. 30 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 22 (50 uM) treated.

FIG. 31 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 22 (5 uM) treated. FIG. 32 shows LQT3 cells recordings of action potentials with solid line representing baseline and dotted line represented compound 23 (50 uM) treated.

FIG. 33 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 23 (50 uM) treated.

FIG. 34 shows LQT1 cells recordings of action potentials carried out in duplicate well on the multiwell plate with solid line representing baseline and dotted line represented compound 23 (5 uM) treated.

FIG. 35 shows recordings of action potentials of control A.

FIG. 36 shows recordings of action potentials of control B.

FIG. 37 shows recordings of action potentials of control C.

DETAILED DESCRIPTION

This disclosure provides compounds useful for treating cardiac arrhyth ia, ventricular arrhythmia, hypertrophic cardiomyopathy, or congestive heart failure, for example, Long QT syndrome (LQTS). Also provided herein are methods and materials for using such compounds to shorten myocardial repolarization time in a patient. For example, a compound provided herein can be used to treat cardiac arrhythmias, such as LQTS (e.g., congenital LQTS and drug induced LQTS). In some cases, a patient is afflicted with a disease or disorder characterized by a prolonged myocardial repolarization time and can be treated with a compound provided herein (e.g., a compound of Formula (I) or a

pharmaceutically acceptable salt thereof). For example, a compound of Formula (I) can be used to shorten myocardial repolarization time in a patient.

The present disclosure provides a compound of Formula (I):

or a pharmaceutically acceptable salt thereof,

wherein: each R ¹ is independently selected from the group consisting of Ci- ₆ alkyl, Ci- ₆ alkoxy, and Ci- ₆ haloalkyl;

L is a bond or selected from the group consisting of C(O), C(0)NH, C(0)NCH ₃ , NHCO, NCH C(0), SO2NH, and NHS0 ₂ ;

A is a bond or a 5-membered heteroaryl; wherein the 5-membered heteroaryl has at least one ring-forming carbon atom and 1, 2, or 3 ring-forming N atoms;

Cy is selected from the group consisting of C6-10 aryl, 5-10 membered heteroaryl, and 4-14 membered heterocycloalkyl; wherein each 5-10 membered heteroaryl and 4-14 membered heterocycloalkyl has at least one ring-forming carbon atom and 1, 2, 3, or 4 ring forming N or S atoms; wherein the one or more ring-forming carbon atoms of the 5-10 membered heteroaryl and 4-14 membered heterocycloalkyl are optionally substituted by oxo to form a carbonyl group; and wherein the C6-10 aryl, 5-10 membered heteroaryl, and 4-14 membered heterocycloalkyl are each substituted with n R ⁶ ;

each R ⁶ is independently selected from the group consisting of Ci- ₆ alkyl, Ci- ₆ alkoxy, Ci- ₆ haloalkyl, and C(0)NH ₂ ;

m is 0, 1, 2, 3, 4 or 5; and

n is 0, 1, 2, 3, 4, or 5;

provided that when L is C(0)NH or NHC(O) and A is a bond, Cy is not unsubstituted or substituted phenyl.

In some embodiments, each R ¹ is independently selected from the group consisting of Ci- ₆ alkyl, Ci- ₆ alkoxy, and Ci- ₆ haloalkyl;

L is a bond or selected from the group consisting of C(O), C(0)NCH ₃ , NCH ₃ C(0), SO2NH, and NHSO2;

A is a bond or a 5-membered heteroaryl; wherein the 5-membered heteroaryl has at least one ring-forming carbon atom and 1, 2, or 3 ring-forming N atoms;

Cy is selected from the group consisting of C6-10 aryl, 5-10 membered heteroaryl, and 4-14 membered heterocycloalkyl; wherein each 5-10 membered heteroaryl and 4-14 membered heterocycloalkyl has at least one ring-forming carbon atom and 1, 2, 3, or 4 ring forming N or S atoms; wherein the one or more ring-forming carbon atoms of the 5-10 membered heteroaryl and 4-14 membered heterocycloalkyl are optionally substituted by oxo to form a carbonyl group; and wherein the C6-10 aryl, 5-10 membered heteroaryl, and 4-14 membered heterocycloalkyl are each substituted with n R ⁶ ; each R ⁶ is independently selected from the group consisting of Ci- ₆ alkyl, Ci- ₆ alkoxy, Ci- ₆ haloalkyl, and C(0)NH ₂ ;

m is 0, 1, 2, 3, 4 or 5; and

n is 0, 1, 2, 3, 4, or 5.

Cy can be selected from the group consisting of phenyl, benzoimidazolyl,

tetrahydroisoquinolinyl, pyridyl, phenothiazinyl, dihydroisoquinolinonyl and isoindolinonyl; wherein the phenyl, benzoimidazolyl, tetrahydroisoquinolinyl, pyridyl, phenothiazinyl, dihydroisoquinolinonyl and isoindolinonyl are each substituted with n R ⁶ . In some embodiments, Cy is phenyl substituted with 1 or 2 R ⁶ . For example, Cy is phenyl substituted with 1 R ⁶ . In some embodiments, Cy is phenyl substituted with 2 R ⁶ . Cy can be unsubstituted benzoimidazolyl or benzoimidazolyl substituted with 1 or 2 R ⁶ . In some examples, Cy can be unsubstituted tetrahydroisoquinolinyl or tetrahydroisoquinolinyl substituted with 1 R ⁶ . Cy can be pyridyl substituted with 1 R ⁶ . Cy can be unsubstituted phenothiazinyl. Cy can be dihydroisoquinolinonyl substituted with 1 R ⁶ . Cy can be isoindolinonyl substituted with 1 R ⁶ .

In some examples, the compound is represented by Formula la:

la,

or a pharmaceutically acceptable salt thereof, wherein Xi and X ₂ are each N or CR ^X , provided both Xi and X ₂ are not both N; R ^x is hydrogen or R ⁶ ; and R ¹ , R ⁶ , L, and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula lb:

lb, or a pharmaceutically acceptable salt thereof, wherein A, R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula Ic:

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula Id:

or a pharmaceutically acceptable salt thereof, wherein Y is CFh, CFh-CFh, or CH-N; and R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula Ial :

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2. In some examples, the compound is represented by Formula Ia2:

Ia2,

or a pharmaceutically acceptable salt thereof, wherein one of Xi and X2 is N and the other of Xi and X2 is CR ^X ; R ^x is hydrogen or R ⁶ ; and R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2. In some examples, Xi is N and X2 is CR ^X . In some examples, Xi is CR ^X and X2 is N.

In some examples, the compound is represented by Formula Ia3 :

Ia3,

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula Ibl :

Ibl,

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula IIb2:

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula IIb3 :

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula Idl :

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula Id2:

M2,

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, the compound is represented by Formula Id3 :

Id3,

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ⁶ , and n are as defined herein. For example, R ¹ can be Ci- ₆ alkoxy. R ⁶ can be Ci- ₆ alkyl or Ci- ₆ haloalkyl. In some examples, R ⁶ is alkyl. In some examples, R ⁶ is haloalkyl. In some examples, n is 1. In other examples, n is 2.

In some examples, R ¹ can be Ci- ₆ alkyl. In some examples, R ¹ is Ci- ₆ alkoxy. In other examples, R ¹ is Ci- ₆ haloalkyl. For example, each R ¹ can be independently methyl, methoxy, or trifluorom ethyl. In some examples, each R ¹ is independently methoxy or trifluorom ethyl.

In some embodiments, R ¹ is methoxy.

Each R ⁶ can be independently Ci- ₆ alkyl, Ci- ₆ alkoxy, or Ci- ₆ haloalkyl. In some examples, each R ⁶ is independently Ci- ₆ alkyl or Ci- ₆ haloalkyl. R ⁶ can be Ci- ₆ alkyl. In some examples, R ⁶ is Ci- ₆ haloalkyl. For example, each R ⁶ can be independently methyl, trifluoromethyl, or C(0)NH ₂ .

In some examples, n is 0, 1 or 2. For example, n is 1.

In some examples, the compound is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

In some examples, the compound is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

Uses of the Compounds The methods provided herein include methods for the treatment or prevention of cardiac arrhythmia, ventricular arrhyth ia, hypertrophic cardiomyopathy, and congestive heart failure. The methods provided herein also include methods for shortening myocardial repolarization time. The patient can have LQTS (e.g., congenital LQTS and drug induced LQTS). For example, the patient can have LQTS type 2. Generally, the methods include administering a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof as described herein (i.e., a compound of Formula (I)), to a patient who is in need of, or who has been determined to be in need of, such treatment.

Effective, efficient ventricular pumping action depends on proper cardiac function. Proper cardiac function, in turn, relies on the synchronized contractions of the heart at regular intervals. When normal cardiac rhythm is initiated at the sinoatrial node, the heart is said to be in sinus rhythm. However, when the heart experiences irregularities in its coordinated contraction, due to electrophysiologic disturbances, e.g., caused by a disease process or from an electrical disturbance, the heart is described as being arrhythmic. The resulting cardiac arrhythmia impairs cardiac efficiency and can be a potentially life threatening event.

Cardiac arrhythmia includes any disorder where there is abnormal electrical activity in the heart. For example, cardiac arrhythmia includes LQTS, premature atrial contractions, wandering atrial pacemaker, multifocal atrial tachycardia, atrial flutter, atrial fibrillation, supraventricular tachycardia, AV nodal reentrant tachycardia, junctional rhythm, junctional tachycardia, premature junctional complex, premature ventricular contractions, accelerated idioventricular rhythm, monomorphic ventricular tachycardia, polymorphic ventricular tachycardia, and ventricular fibrillation. In some embodiments, the cardiac arrhythmia is LQTS.

LQTS is characterized by a prolongation of the myocardial repolarization time and can either be congenital or acquired as a result of medication or a metabolic disturbance (e.g., drug induced LQTS).

Congenital LQTS can arise from a mutation of one of several genes. These mutations tend to prolong the ventricular action potential duration (APD), thus lengthening the QT interval. LQTS can be inherited in an autosomal dominant or an autosomal recessive fashion. The autosomal recessive forms of LQTS tend to have a more severe phenotype, with some variants having associated syndactyly (LQT8) or congenital neural deafness (LQT1). LQTS includes, for example, LQT1, LQT2, LQT3, LQT4, LQT5, LQT6, LQT7, LQT8, LQT9, LQT10, LQT11, LQT12, LQT13, Jervell and Lange-Nielsen syndrome (JLNS), and Romano- Ward syndrome. In some embodiments, the LQTS is LQT2.

Drug induced LQTS can be triggered by any drug that triggers a prolongation of myocardial repolarization time in a patient. Such drugs include, for example, Albuterol, Alfuzosin, Amantadine, Amiodarone, Amiodarone, Amitriptyline, Amphetamine, Arsenic trioxide, Astemizole, Atazanavir, Atomoxetine, Azithromycin, Bepridil, Chloral hydrate, Chloroquine, Chlorpromazine, Ciprofloxacin, Cisapride, Citalopram, Clarithromycin, Clomipramine, Clozapine, Cocaine, Desipramine, Dexmethylphenidate, Diphenhydramine, Diphenhydramine, Disopyramide, Dobutamine, Dofetilide, Dolasetron, Domperidone, Dopamine, Doxepin, Dronedarone, Droperidol, Ephedrine, Epinephrine, Erythromycin, Escitalopram, Escitalopram, Famotidine, Felbamate, Fenfluramine, Flecainide, Fluconazole, Fluoxetine, Foscamet, Fosphenytoin, Galantamine, Gatifloxacin, Gemifloxacin, Granisetron, Halofantrine, Haloperidol, Ibutilide, Imipramine, Indapamide, Isoproterenol, Isoproterenol, Isradipine, Itraconazole, Ketoconazole, Lapatinib, Lapatinib, Levalbuterol, Levofloxacin, Levomethadyl, Lisdexamfetamine, Lithium, Mesoridazine, Metaproterenol, Methadone, Methylphenidate, Midodrine, Moexipril/HCTZ, Moxifloxacin, Nicardipine, Nilotinib, Norepinephrine, Nortriptyline, Octreotide, Ofloxacin, Ondansetron, Oxytocin, Paliperidone, Paroxetine, Pentamidine, Perflutren lipid microspheres, Phentermine, Phenylephrine, Phenylpropanolamine, Pimozide, Probucol, Procainamide, Protriptyline, Pseudoephedrine, Quetiapine, Quinidine, Ranolazine, Risperidone, Ritodrine, Ritonavir, Roxithromycin, Salmeterol, Sertindole, Sertraline, Sibutramine, Solifenacin, Sotalol, Sparfloxacin, Sunitinib, Tacrolimus, Tamoxifen, Telithromycin, Terbutaline, Terfenadine, Thioridazine, Tizanidine, Tolterodine, Trazodone, Trimethoprim-Sulfa, Trimipramine, Vandetanib, Vardenafil, Venlafaxine, Voriconazole, Ziprasidone, and combinations thereof.

Without being bound by theory, it is believed that the compounds of Formula (I) inhibit late sodium current to shorten ventricular APD (i.e. myocardial repolarization time). Accordingly, a compound of Formula (I) can function as a selective inhibitor of the late sodium current relative to peak sodium channel current, and via this mechanism, it may decrease sodium-dependent intracellular calcium overload. See, for example, (Scirica BM, et al. Circulation. 2007; 116(15): 1647-52 and Morrow DA, et al. JAMA 2007; 297(16): 1775- 83). The methods provided herein also include methods for the treatment of disorders associated with sodium-dependent intracellular calcium overload. In some embodiments, the disorder is selected from coronary artery disease and atrial fibrillation.

The treatment provided herein can ameliorate at least one symptom of cardiac arrhythmia, ventricular arrhyth ia, hypertrophic cardiomyopathy, or congestive heart failure. The symptoms can include fluttering in the chest, irregular heartbeats, chest pain, shortness of breath, dizziness, sweating, fainting, heart murmur, congested lungs, fluid and water retention, seizure, etc. Cardiac arrhythmias can result in prolonged myocardial repolarization time; thus, a treatment can result in the shortening of myocardial repolarization time and a return or approach to a regular cardiac rhythm. In some embodiments, administration of a therapeutically effective amount of a compound described herein for the treatment of a disorder, disease, or condition can result in a shortened myocardial repolarization time. In some instances, LQTS can lead to sudden death, and administration of a therapeutically effective amount of a compound described herein can be used in resuscitation.

Depending on the patient being treated, the compounds provided herein may be formulated and administered acutely or chronically. As used herein,“acute” administration refers to administration of one or a few doses to a patient, typically at or near onset of symptoms, diagnosis of arrhythmia, or presentation for treatment, e.g., within 24 hours. In some embodiments, acute administration can be used to treat a subject who is presently experiencing arrhythmia.

As used herein,“chronic” administration refers to administration of more than one dose to a patient, wherein the doses are administered over a longer period of time; typically each dose is administered before the previous dose is completely cleared from the patient. In some embodiment, chronic administration can include daily administration, e.g., for a week, two weeks, a month or more, e.g., to a patient who has previously experienced arrhythmia, or is at high risk of experiencing an arrhyth ia (e.g., due to personal or family history, or genetic or environmental factors;“high” risk refers to a risk above that of a normal, healthy member of the general population). Chronic administration can be used to reduce a subject’s risk of experiencing a cardiac arrhythmia, e.g., in a subject who is at high risk of experiencing a cardiac arrhythmia. In some embodiments, a compound of Formula (I) as described herein is administered chronically.

In some embodiments, one or more of the compounds provided herein may be administered to a patient in the methods provided herein. For example, two compounds of Formula (I) can be administered chronically to prevent (e.g., reduce the risk of) or treat recurring a disorder in a patient. One or more of the compounds provided herein can be administered to the patient in combination. The compounds can be administered together or administration of one may precede administration of the other.

The methods provided herein include the manufacture and use of pharmaceutical compositions, which include compounds provided herein, e.g., compounds of Formula (I), or a pharmaceutically acceptable salt thereof. Also included are the pharmaceutical compositions themselves.

Pharmaceutical compositions typically include a pharmaceutically acceptable carrier. As used herein the language“pharmaceutically acceptable carrier” includes saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

A pharmaceutical composition is typically formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal, and rectal administration.

Methods of formulating suitable pharmaceutical compositions are known in the art, see, e.g., Remington: The Science and Practice of Pharmacy, 21st ed., 2005; and the books in the series Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs (Dekker, NY). For example, solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol, or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates, or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes, or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injection can include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. The composition should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyetheylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, and sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle, which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying, which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an edible carrier. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules, e.g., gelatin capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash.

Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds can be delivered in the form of an aerosol spray from a pressured container or dispenser that contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer. Such methods include those described in U.S. Patent No. 6,468,798.

Systemic administration of a therapeutic compound as described herein can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal

administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.

The pharmaceutical compositions can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

Additionally, intranasal delivery is possible, as described in, inter alia, Hamajima et ak, Clin. Immunol. Immunopathol. , 88(2), 205-10 (1998). Liposomes (e.g., as described in U.S. Patent No. 6,472,375) and microencapsulation can also be used. Biodegradable targetable microparticle delivery systems can also be used (e.g., as described in U.S. Patent No. 6,471,996).

In one embodiment, the therapeutic compounds are prepared with carriers that will protect the therapeutic compounds against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Such formulations can be prepared using standard techniques, or obtained commercially, e.g., from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to selected cells with monoclonal antibodies to cellular antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Patent No. 4,522,811.

The pharmaceutical composition may be administered at once, or may be divided into a number of smaller doses to be administered at intervals of time. It is understood that the precise dosage and duration of treatment is a function of the disease being treated and may be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test data. It is to be noted that concentrations and dosage values may also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular patient, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed

compositions.

Dosage forms or compositions containing a compound as described herein in the range of 0.005% to 100% with the balance made up from non-toxic carrier may be prepared. Methods for preparation of these compositions are known to those skilled in the art. The contemplated compositions may contain 0.001%-100% active ingredient, in one embodiment 0.1-95%, in another embodiment 75-85%.

The pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.

Synthesis

Compounds and salts thereof described herein can be prepared using known synthetic techniques. The compounds can be synthesized according to any of various possible synthetic routes, such as those in the schemes below. The reactions can be carried out in suitable solvents which can be readily selected by a person skilled in the art. Suitable solvents includes non-reactive with the starting materials (reactants), the intermediates or products at the temperatures at which the reactions are carried out. A particular reaction can be carried out in one solvent or a mixture of more than one solvent.

Reactions can be monitored according to any suitable methods known in the art. For example, product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy ( e.g ., 'H or ¹³ C), infrared spectroscopy, spectrophotometry ( e.g ., UV-visible), mass spectrometry or by chromatographic methods such as high performance liquid chromatography (HPLC) or thin layer chromatography (TLC).

The schemes below provide general guidance in connection with preparing the compounds described herein. One skilled in the art would understand that the preparations shown in the schemes can be modified or optimized using general knowledge of organic chemistry to prepare various compounds described herein.

For example, the compounds can be prepared according to Scheme 1.

Scheme 1

L' = S0 ₂ , CO

X = C1

Compound A1 is reacted with Compound A2 in the presence of one or more solvents, such as dichloromethane, and a base, such as trimethylamine and pyridine.

In some examples, compounds provided herein can be prepared according to Scheme

2

Scheme 2

X = I

Compound B1 is coupled to compound B2 in the presence of sodium azide and copper iodide and one or more solvents, such as dimethylsulfoxide and water.

In some examples, compounds of this disclosure can be prepared according to Scheme 3.

Scheme 3

Compound Cl is converted to Compound C2 in the presence of an amine (e.g., CH3NH2, NH3) in an alcohol solvent. Compound C2 can be reduced in the presence of a reducing agent, such as Raney N1/H2, to generate Compound C3, which can be coupled with

Compound C4 to form Compound C5. Compound C5 can be treated with an acid, such as acetic acid, to generate Compound Ic.

Certain compounds disclosed herein can also be prepared according to Scheme 4.

Scheme 4

Compound D1 can be converted to Compound D2 in the presence of an azide (e.g., sodium azide) and an acid (e.g., hydrochloric acid). Compound D2 can be coupled with a halobenzene (e.g., iodobenzene) in the presence of a copper catalyst to afford Compound Id.

Certain compounds described herein can be prepared according to the Scheme 5 below.

Scheme 5

Compound D3 can be converted to Compound D4 in the presence of e.g., SOCh and methanol. Compound D4 can be treated with N-bromosuccinimide, azobisisobutyronitrile, and carbon tetrachloride to generate D5, which can be coupled with D6 to form Compound Id.

Definitions

For the terms“for example” and“such as,” and grammatical equivalences thereof, the phrase“and without limitation” is understood to follow unless explicitly stated otherwise. As used herein, the term“about” is meant to account for variations due to experimental error.

As used herein, the singular forms“a,”“an,” and“the” include plural referents unless the context clearly dictates otherwise.

A“patient,” as used herein, includes both humans and other animals, particularly mammals. Thus, the methods are applicable to both human therapy and veterinary applications. In some embodiments, the patient is a mammal, for example, a primate. In some embodiments, the patient is a human.

A“therapeutically effective” amount of a compound provided herein is typically one which is sufficient to achieve the desired effect and may vary according to the nature and severity of the disease condition, and the potency of the compound. It will be appreciated that different concentrations may be employed for prophylaxis than for treatment of an active disease.

The term,“compound,” as used herein is meant to include all stereoisomers, geometric isomers, and tautomers of the structures depicted. Compounds herein identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise specified.

In some embodiments, a compound provided herein, or salt thereof, is substantially isolated. By“substantially isolated” is meant that the compound is at least partially or substantially separated from the environment in which it was formed or detected. Partial separation can include, for example, a composition enriched in the compound provided herein. Substantial separation can include compositions containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the compound provided herein, or salt thereof. Methods for isolating compounds and their salts are routine in the art.

The phrase“pharmaceutically acceptable” is used herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication,

commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salts" refers to when the compounds provided herein are converted to their salt forms. Examples of pharmaceutically acceptable salts include inorganic or organic acid salts. The pharmaceutically acceptable salts can be synthesized from basic or acidic moiety of the compound provided herein by conventional chemical methods. The nanoprobe provided herein can be positively charged and anions can be associated with the charged nanoprobe to form a salt. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17 ^th Ed., (Mack Publishing Company, Easton, 1985), p. 1418, Berge et al., ./. Pharm. Sci., 1977, 66( 1), 1-19 and in Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use , (Wiley, 2002).

The term“alkyl” includes straight-chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl) and branched-chain alkyl groups (isopropyl, /f/V-butyl, isobutyl, and sec-butyl), cycloalkyl (alicyclic) groups (cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl), alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups. In certain embodiments, a straight chain or branched chain alkyl has six or fewer carbon atoms in its backbone (e.g., C 1-C6 for straight chain; C3- C ₆ for branched chain). The term C 1-C6 includes alkyl groups containing 1 to 6 carbon atoms.

As used herein,“aryl” refers to monocyclic or polycyclic (e.g., having 2, 3 or 4 fused rings) aromatic hydrocarbons such as, for example, phenyl, naphthyl, anthracenyl, and phenanthrenyl. In some embodiments, aryl groups have from 6 to about 20 carbon atoms. As used herein,“heteroaryl” groups refer to aromatic heterocycles having at least one heteroatom ring member such as sulfur, oxygen, or nitrogen. Heteroaryl groups include monocyclic and polycyclic (e.g., having 2, 3 or 4 fused rings) systems. Examples of heteroaryl groups include without limitation, pyridyl, N-oxopyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furyl, quinolyl, isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl, pyrryl, oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl, isothiazolyl, benzothienyl, purinyl, carbazolyl, benzimidazolyl, and indolinyl. In some embodiments, the heteroaryl group has from 1 to about 20 carbon atoms. In some embodiments, the heteroaryl group contains 3 to about 14 ring-forming atoms. In other embodiments, the heteroaryl group has 1 to about 4

heteroatoms.

As used herein, the term“heterocycloalkyl” refers to non-aromatic heterocycles having at least one heteroatom ring member such as sulfur, oxygen, or nitrogen.

Heterocycloalkyl can include 4-10 ring members, 4-7 ring members, or 4-6 ring members. Heterocycloalkyl groups can include mono- or bicyclic or spirocyclic ring systems. Ring forming carbon atoms and heteroatoms of a heterocycloalkyl group can be optionally oxidized to form an oxo or sulfido group or other oxidized linkage (e.g., C(O), S(O), C(S) or S(0) ₂ , A-oxide etc.) or a nitrogen atom can be quaternized. The heterocycloalkyl group can be attached through a ring-forming carbon atom or a ring-forming heteroatom.

Heterocycloalkyl also include groups that have one or more aromatic rings fused to the heterocycloalkyl ring, e.g., benzo or thienyl derivatives of piperidine, morpholine, azepine, etc.

The term“halo” includes chloro, bromo, iodo, and fluoro.

The term“haloalkyl” refers to an alkyl as defined above having one or more its hydrogen replaced with a halogen.

The term "alkoxy," refers to a group of formula -O-alkyl, wherein the alkyl group is as defined above. Example alkoxy groups include methoxy, ethoxy, propoxy, /-butoxy, etc.

The terms“treat,”“treating,” or“treatment,” refer to reversing, inhibiting, or alleviating the disease, condition, or disorder in a subject who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder.

The terms“prevent,”“preventing,” or“prevention,” refer to reducing the risk of having a disease, condition, or disorder in a subject who may be predisposed to the disease, condition or disorder but does not yet experience or display the pathology or symptomatology of the disease, condition or disorder. For example, the compounds described herein can be administered to a patient who does not have any symptoms of a disease (e.g., cardiac arrhythmia, ventricular arrhyth ia, hypertrophic cardiomyopathy, or congestive heart failure), and such administration can prevent the onset or activation of the disease. The prevention can last for days, months, or years after the administration. In some cases, the patient may be predisposed for the disease either due to e.g., hereditary conditions or lifestyle factors. Administration of a compound described herein can also reduce the risk of having the disease in these predisposed patients.

It is further appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, can also be provided in combination in a single embodiment (while the embodiments are intended to be combined as if written in multiply dependent form). Conversely, various features of the invention which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable subcombination. Thus, it is contemplated as features described as

embodiments of the compounds of Formula (I) can be combined in any suitable combination.

EXAMPLES

The following examples are illustrative and not limiting. Example 1. 2-m ethoxy-N-(6-methylpyri din-3 -yl)benzamide (3)

To a solution of 6-methyl pyri din-3 -amine 1 (108 mg, 1 mmol) and triethylamine (202 mg, 2 mmol) in dichloromethane (10 mL) was added 2-methoxybenzoyl chloride 2 (170 mg,

1 mmol) at 0 °C, and the mixture stirred at 0 °C for 1 hour. The resulting mixture was washed with water (2 x 50 mL), brine (2 x 50 mL), dried over anhydrous sodium sulfate, and filtered through a pad of diatomaceous earth. The filtrate was concentrated in vacuo to afford a residue. The residue was purified by preparative thin-layer chromatography eluting with 2:3 petroleum ether/ethyl acetate to afford 2-methoxy-N-(6-methylpyridin-3-yl)benzamide 3 as a white solid (140 mg, 58%). LCMS analysis of 3: 100% purity and [M+H] ⁺ = 243. Example 2. 2-methoxy-N-(5-methylpyridin-2-yl)benzamide (6)

A mixture of 5-methylpyridin-2-amine 4 (108 mg, 1 mmol), 2-methoxybenzoic acid 5 (152 mg, 1 mmol), (l-[bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5-b]pyr idinium 3- oxide hexafluorophosphate) (570 mg, 1.5 mmol), and triethylamine (202 mg, 2 mmol) in di chi orom ethane (10 mL) was stirred at room temperature for 16 hours. The resulting mixture was concentrated in vacuo to afford a residue. The residue was purified by preparative thin-layer chromatography eluting with 1 : 1 petroleum ether/ethyl acetate to afford 2-methoxy-N-(5-methylpyridin-2-yl)benzamide 6 as a white solid (124 mg, 57%). LCMS analysis of 6: 100% and [M+H] ⁺ = 243.

Example 3. 2-(2-methoxyphenyl)-l-methyl-7-(trifluoromethyl)-lH-benzo[d] imidazole (11)

10 11 Preparation of N-methyl-2-nitro-6-(trifluoromethyl)aniline (8)

2-fluoro-l-nitro-3 -(triflu orom ethyl )benzene 7 (1.1 g, 5.26 mmol) in a methylamine alcohol solution (5 mL) was stirred at room temperature for 2 hours. The resulting mixture was concentrated in vacuo to afford a residue. The residue was purified by flash

chromatography over silica gel eluting with 10: 1 petroleum ether/ethyl acetate to afford N- methyl-2-nitro-6-(trifluoromethyl)aniline 8 as a yellow oil (1.1 g, 95%). LCMS analysis of 6: 100% and [M+H] ⁺ = 221.

Preparation of N1 -methyl -6-(trifluoromethyl)benzene-l, 2-diamine (9)

N-methyl-2-nitro-6-(trifluoromethyl)aniline 8 (1.1 g, 5 mmol) and Raney Nickel (1 g) were stirred in methanol (10 mL) under hydrogen at room temperature for 2 hours. The resulting mixture was filtered and concentrated in vacuo to afford crude Nl-methyl-6- (trifluorom ethyl )benzene- 1,2-diamine 9 as a brown oil (800 mg, 84%). LCMS analysis of 9: 63% and [M+H] ⁺ = 191.

Preparation of 2-methoxy-N-(2-(methylamino)-3-(trifluoromethyl)phenyl)benza mide (10)

To a solution of Nl-methyl-6-(trifluoromethyl)benzene-l, 2-diamine 9 (800 mg, 4.2 mmol) and triethylamine (850 mg, 8.4 mmol) in dichloromethane (20 mL) was added 2- methoxybenzoyl chloride 2 (785 mg, 4.62 mmol) dropwise at 0 °C, then stirred at 0 °C for 1 hour. The resulting mixture was washed with water (2 x 50 mL), brine (2 x 50 mL), dried over anhydrous sodium sulfate, and filtered through a pad of diatomaceous earth. The filtrate was concentrated in vacuo to afford crude 2-methoxy-N-(2-(methylamino)-3- (trifluoromethyl)phenyl)benzamide 10 as a yellow oil which was used as is in the next step (1.5 g, 80%). LCMS analysis of 10: 63% and [M+H] ⁺ = 325.

Preparation of 2-(2-methoxyphenyl)-l-methyl-7-(trifluoromethyl)-lH-benzo[d] imidazole

(11)

A solution of 2-methoxy-N-(2-(methylamino)-3 -(trifluorom ethyl )phenyl)benzamide 10 (1.3 g, 4 mmol) in acetic acid (10 mL) was stirred at 80 °C for 2 hours. The resulting mixture was concentrated in vacuo to provide a material which was taken up in ethyl acetate (50 mL) and saturated aqueous sodium hydrogen carbonate (100 mL) and stirred for 10 minutes. The layers were partitioned, and the aqueous layer partitioned with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered through a pad of diatomaceous earth and concentrated in vacuo to provide a residue. The residue was purified by preparative thin-layer

chromatography eluting with 1 :3 petroleum ether/dichl orom ethane to afford 2-(2- methoxyphenyl)-l-methyl-7-(trifluoromethyl)-lH-benzo[d]imida zole 11 as a white solid (350 mg, 35%). LCMS analysis of 11: 100%, and [M+H] ⁺ = 307.

Example 4. 2-(2-methoxyphenyl)-7-(trifluoromethyl)-lH-benzo[d]imidazole (15)

Preparation of 2-nitro-6-(trifluoromethyl)aniline (12)

2-fluoro-l-nitro-3 -(triflu orom ethyl )benzene 7 (1.1 g, 5.26 mmol) in a solution of ammonia in methanol (7N, 10 mL) was stirred at 70 °C for 16 hours. The resulting mixture was concentrated in vacuo to afford a residue. The residue was purified by flash

chromatography over silica gel eluting with 10: 1 petroleum ether/ethyl acetate to afford 2- nitro-6-(trifluoromethyl)aniline 12 as a yellow oil (1.0 g, 90%). LCMS analysis of 12:

97.4%, and [M+H] ⁺ = 207.

Preparation of 3 -(trifluoromethyl)benzene- 1,2-diamine (13) 2-nitro-6-(trifluoromethyl)aniline 12 (1.0 g, 4.85 mmol) and Raney Nickel (1 g) were stirred in methanol (30 mL) under hydrogen at room temperature for 2 hours. The resulting mixture was filtered and concentrated in vacuo to afford crude 3-(trifluoromethyl)benzene- 1, 2-diamine 13 as a brown oil which was used as is in the next step (850 mg, 95%). LCMS analysis of 13: 97% and [M+H] ⁺ = 177.

Preparation of N-(2-amino-3-(trifluoromethyl)phenyl)-2-methoxybenzamide (14)

To a solution of 3-(trifluoromethyl)benzene-l, 2-diamine 13 (850 mg, 4.83 mmol) and triethylamine (1 g, 9.66 mmol) in dichloromethane (20 mL) was added 2-methoxybenzoyl chloride 2 (820 mg, 4.83 mmol) dropwise at 0 °C, then stirred at 0 °C for 1 hour. The resulting mixture was washed with water (2 x 20 mL), brine (2 x 20 mL), dried over anhydrous sodium sulfate, and filtered through a pad of diatomaceous earth. The filtrate was concentrated in vacuo to afford crude N-(2-amino-3-(trifluorom ethyl )phenyl)-2- methoxybenzamide 14 as a yellow oil which was used as is in the next step (1.45 g, 85%). LCMS analysis of 14: 96% and [M+H] ⁺ = 311.

Preparation of 2-(2-methoxyphenyl)-7-(trifluoromethyl)-lH-benzo[d]imidazole (15)

A solution of N-(2-amino-3-(trifluoromethyl)phenyl)-2-methoxybenzamide 14 (1.45 g, 4.68 mmol) in acetic acid (10 mL) was stirred at 80 °C for 2 hours. The resulting mixture was concentrated in vacuo and purified by flash chromatography over silica gel eluting with 20: 1 petroleum ether/dichloromethane to afford 2-(2-methoxyphenyl)-7-(trifluoromethyl)- lH-benzo[d]imidazole 15 as a white solid (250 mg, 17%). LCMS analysis of 15: 100% and [M+H] ⁺ = 293.

Example 5. 2-(2-methoxyphenyl)-5-(p-tolyl)-lH-imidazole (19)

16 17 19 Preparation of 2-methoxybenzimidamide (17)

To ammonium chloride (2.16 g, 40 mmol) in toluene (80 mL) was added aluminum chloride (2N in toluene, 20 mL, 40 mmol) dropwise under nitrogen at 0 °C. After addition was complete, the mixture was stirred to room temperature for another 30 minutes. To the resulting mixture was added 2-methoxybenzonitrile 16 (5.32 g, 40 mmol) in toluene (20 mL) slowly, then stirred at 80 °C for 16 hours. The reaction mixture was cooled to room temperature and poured into a slurry of silica gel (100 g) in chloroform (200 mL). The mixture was stirred for 10 minutes and filtered. The residue was washed with methanol (200 mL) and the combined filtrates concentrated in vacuo. To the resulting residue was added dichloromethane (20 mL), and the mixture filtered to provide 2-methoxybenzimidamide 17 as a white solid (1.2 g, 20%). LCMS analysis of 17: 100% and [M+H]+ = 151.

Preparation of 2-(2-methoxyphenyl)-5-(p-tolyl)-lH-imidazole (19)

To 2-methoxybenzimidamide 17 (150 mg, 1 mmol) and potassium carbonate (414 mg, 3 mmol) in acetonitrile (30 mL) was added 2-bromo-l-(p-tolyl)ethan-l-one 18 (212 mg,

1 mmol) in acetonitrile (5 mL) dropwise at 0 °C. The mixture was allowed to slowly equilibrate to room temperature and stirred for 16 hours. The reaction mixture was filtered and concentrated in vacuo. The resulting residue was purified by preparative thin-layer chromatography eluting with dichloromethane to afford 2-(2-methoxyphenyl)-5-(p-tolyl)-lH- imidazole 19 as a white solid (70 mg, 27%). LCMS analysis of 19: 100% and [M+H]+ =

265.

Example 6. 2-(2-methoxyphenyl)-5-(2-(trifluoromethyl)phenyl)-lH-imidazo le (21)

To 2-methoxybenzimidamide 17 (150 mg, 1 mmol) and potassium carbonate (414 mg, 3 mmol) in acetonitrile (30 mL) was added 2-bromo-l-(2-(trifluorom ethyl )phenyl)ethan- 1-one 20 (266 mg, 1 mmol) in acetonitrile (10 mL) dropwise at 0 °C. The mixture was stirred an additional 2 hours at 0 °C, then filtered and concentrated in vacuo. Acetic acid (20 mL) was added to the resulting residue, which was stirred at 60 °C overnight. The mixture was concentrated in vacuo to provide a residue that was taken up in saturated aqueous sodium hydrogen carbonate (30 mL) and extracted with ethyl acetate (2 x 30 mL). The combined organic layers were washed with brine (20 mL) and dried over anhydrous sodium sulfate, then concentrated in vacuo to provide a residue that was purified by preparative thin-layer chromatography eluting with dichloromethane to afford 2-(2-methoxyphenyl)-5-(2- (trifluoromethyl)phenyl)-lH-imidazole 21 as a white solid (130 mg, 41%). LCMS analysis of 19: 100% and [M+H]+ = 319.

Example 7. N-benzhydryl-2-methoxybenzamide (23)

To a solution of 2-methoxybenzoyl chloride 2 (200 mg, 1.17 mmol) in

dichloromethane (15 mL) were added diphenylmethanamine 22 (280 mg, 1.53 mmol) and triethylamine (356 mg, 3.52 mmol) at room temperature, then stirred for 16 hours. The mixture was concentrated in vacuo and purified by flash chromatography over silica gel eluting over a gradient of 94:6 to 96:4 petroleum ether/ethyl acetate to afford N-benzhydryl- 2-methoxybenzamide 23 (250 mg). LCMS analysis of 23: [M+H] ⁺ = 318. Example 8. (2-methoxyphenyl)(10H-phenothiazin-10-yl)methanone (25)

To a solution of 2-methoxybenzoyl chloride 2 (200 mg, 1.17 mmol) in

dichloromethane (15 mL) were added 10H-phenothiazine 24 (304 mg, 1.53 mmol) and triethylamine (356 mg, 3.52 mmol) at room temperature, then stirred for 16 hours. The mixture was concentrated in vacuo and purified by flash chromatography over silica gel eluting over a gradient of 94:6 to 96:4 petroleum ether/ethyl acetate to afford (2- methoxyphenyl)(10H-phenothiazin-10-yl)methanone 25 (120 mg). LCMS analysis of 25:

[M+H] ⁺ = 334.

Example 9. 2-methoxy-N-methyl-N-(2-(trifluoromethyl)phenyl)benzamide (27)

c

2

A mixture of 2-methoxybenzoyl chloride 2 (200 mg, 1.17 mmol) and N-methyl-2- (trifluoromethyl)aniline 26 (267 mg, 1.53 mmol) in xylene (2 mL) was stirred at 140 °C for 16 hours. The mixture was concentrated in vacuo and purified by flash chromatography over silica gel eluting over a gradient of 8: 1 to 4: 1 petroleum ether/ethyl acetate to afford 2- methoxy-N-methyl-N-(2-(trifluoromethyl)phenyl)benzamide 27 (70 mg). LCMS analysis of 27: [M+H] ⁺ = 310.

Example 10. 2-methoxy-N-(4-methyl-2-(trifluoromethyl)phenyl)benzamide (29)

A mixture of 2-methoxybenzoyl chloride 2 (200 mg, 1.17 mmol) and 4-m ethyl-2- (trifluoromethyl)aniline 28 (350 mg, 1.53 mmol) in xylene (2 mL) was stirred at 140 °C for 16 hours. The mixture was concentrated in vacuo and purified by flash chromatography over silica gel eluting over a gradient of 30: 1 to 20: 1 petroleum ether/ethyl acetate to afford 2- methoxy-N-(4-methyl-2-(trifluoromethyl)phenyl)benzamide 29 (100 mg). LCMS analysis of 29: [M+H] ⁺ = 310. Example 11. N-(2-carbamoylphenyl)-2-methoxybenzamide (31)

To a solution of 2-methoxybenzoyl chloride 2 (200 mg, 1.17 mmol) in

dichloromethane (15 mL) were added 2-aminobenzamide 30 (208 mg, 1.53 mmol) and pyridine (286 mg, 3.52 mmol) and the resulting mixture stirred at room temperature for 16 hours. The mixture was diluted with water (100 mL) and filtered. The solid residue was washed with water (30 mL) and dichloromethane (20 mL), then dried to afford N-(2- carbamoylphenyl)-2-methoxybenzamide 31 (120 mg). LCMS analysis of 31: [M+H] ⁺ = 271.

Example 12. 2-methoxy-N-(2-(trifluoromethyl)phenyl)benzenesulfonamide (34)

Method 1: To a solution of 2-methoxybenzenesulfonyl chloride 32 (1 g, 5.88 mmol) in toluene (20 mL) were added 2-(trifluoromethyl)aniline 33 (1.23 g, 7.65 mmol), pyridine (1.39 g, 17.6 mmol), and 4-(N,N-dimethylamino)pyridine (358 mg, 2.94 mmol), and the resulting mixture stirred at 50 °C for 48 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by flash chromatography over silica gel eluting over a gradient of 9: 1 to 6: 1 petroleum ether/ethyl acetate to afford 2-methoxy-N-(2- (trifluoromethyl)phenyl)benzenesulfonamide 34 (450 mg). LCMS analysis of 34: [M+H] ⁺ = 332.

Method 2: A mixture of 2-methoxybenzenesulfonyl chloride 32 (206 mg, 1 mmol), 2- (trifluoromethyl)aniline 33 (161 mg, 1 mmol), pyridine (202 mg, 2.5 mmol), and

di chi orom ethane (10 mL) were stirred at 35 °C for 48 hours. The mixture was concentrated in vacuo and purified by preparative thin-layer chromatography eluting with 1 : 1 petroleum ether/ethyl acetate to afford 2-methoxy-N-(2-(trifluoromethyl)phenyl)benzenesulfonamide 34 (100 mg). LCMS analysis of 34: 100% and [M+H] ⁺ = 332.

Example 13. 2-methoxy-N-(p-tolyl)benzenesulfonamide (36)

A mixture of 2-methoxybenzenesulfonyl chloride 32 (206 mg, 1 mmol), p-toluidine 35 (112 mg, 1.05 mmol), triethylamine (278 mg, 2 mmol), and dichloromethane were stirred at room temperature for 18 hours. The reaction mixture was diluted with a saturated aqueous solution of sodium hydrogen carbonate (20 mL) and extracted with dichloromethane (3 x 20 mL). The combined organic layers were concentrated in vacuo and purified by preparative thin-layer chromatography eluting with 1 :2 petroleum ether/dichloromethane to afford 2- methoxy-N-(p-tolyl)benzenesulfonamide 36 as a white solid (100 mg). LCMS analysis of 36: 100% and [M+H] ⁺ = 278.

Example 14. 2,6-dimethoxy-N-(2-(trifluoromethyl)phenyl)benzamide (38)

A mixture of 2,6-dimethoxybenzoyl chloride 37 (200 mg, 1 mmol), 2- (trifluoromethyl)aniline 33 (161 mg, 1 mmol), triethylamine (202 mg, 2 mmol), and xylene (10 mL), and the resulting mixture stirred at 140 °C for 16 hours. The mixture was concentrated in vacuo. The resulting residue was purified by preparative thin-layer chromatography eluting with 3: 1 petroleum ether/ethyl acetate to afford 2,6-dimethoxy-N-(2- (trifluoromethyl)phenyl)benzamide 38 (200 mg, 61%) as a brown solid. LCMS analysis of 38: 100% and [M+H] ⁺ = 326.

Example 15. (2-methoxyphenyl)(6-m ethyl-3, 4-dihydroquinolin-l(2H)-yl)methanone (40)

2 40

A mixture of 2-methoxybenzoyl chloride 2 (170 mg, 1 mmol), 6-methyl-l, 2,3,4- tetrahydroquinoline 39 (147 mg, 1 mmol, triethylamine (202 mg, 2 mmol), and

dichloromethane (10 mL) were stirred at room temperature for 16 hours. The mixture was concentrated in vacuo and purified by preparative thin-layer chromatography eluting with 2: 1 petroleum ether/ethyl acetate to afford (2-methoxyphenyl)(6-m ethyl-3, 4-dihy droquinolin- l(2H)-yl)methanone 40 as a brown oil (150 mg, 53%). LCMS analysis of 40: [M+H] ⁺ = 282.

Example 16. (3,4-dihydroisoquinolin-2(lH)-yl)(2-methoxyphenyl)methanone (42)

2 42

A mixture of 2-methoxybenzoyl chloride 2 (340 mg, 2 mmol), 1, 2,3,4- tetrahydroisoquinoline 41 (266 mg, 2 mmol), triethylamine (404 mg, 4 mmol), and dichloromethane (15 mL) were stirred at room temperature for 15 hours. The mixture was concentrated in vacuo and purified by preparative thin-layer chromatography eluting with 3 : 1 petroleum ether/ethyl acetate to afford (3,4-dihydroisoquinolin-2(lH)-yl)(2- methoxyphenyl)methanone 42 as a brown oil (280 mg, 52%). LCMS analysis of 42: 97% and [M+H] ⁺ = 268. Example 17. 7-methoxy-2-(p-tolyl)isoindolin-l-one (46)

Preparation of methyl 2-methoxy-6-methylbenzoate (44)

To a solution of 2-methoxy-6-methylbenzoic acid 43 (1 g, 6 mmol) in methanol (20 mL) was added thionyl chloride (10 mL) at 0 °C. The mixture was then stirred at 85 °C for 16 hours. The mixture was concentrated in vacuo and basified with a saturated aqueous solution of sodium hydrogen carbonate (50 mL). The mixture was stirred for 15 minutes, then extracted with ethyl acetate (3 x 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford methyl 2-methoxy-6- methylbenzoate 44 as a brown oil (500 mg, 46%). LCMS analysis of 44: 90% and [M+H] ⁺ = 181.

Preparation of methyl 2-(bromomethyl)-6-methoxybenzoate (45)

To a round-bottomed flask were added methyl 2-methoxy-6-methylbenzoate 44 (500 mg, 2.8 mmol), N-bromosuccinimide (590 mg, 3.3 mmol), azobisisobutyronitrile (28 mg,

0.15 mmol), and carbon tetrachloride (10 mL) and the resulting mixture stirred at 100 °C for

16 hours. The mixture was cooled to room temperature and filtered. The filtrate concentrated in vacuo to afford methyl 2-(bromomethyl)-6-methoxybenzoate 45 as a brown oil (600 mg, 83%). LCMS analysis of 45: 78% and [M] ⁺ = 259, [M+2] ⁺ = 261.

Preparation of 7-methoxy-2-(p-tolyl)isoindolin-l-one (46)

To a round-bottomed flask were added methyl 2-(bromomethyl)-6-methoxybenzoate 45 (390 mg, 1.5 mmol), p-toluidine 35 (245 mg, 1.5 mmol), acetonitrile (20 mL), and potassium carbonate (420 mg, 3 mmol) and the mixture stirred at 65 °C for 2 hours. The reaction mixture was filtered and the filtrate was treated with sodium methoxide (35% w/w solution in methanol, 10 mL) and methanol (10 mL). The resulting mixture was stirred at 55 °C for 15 minutes, then diluted with water (30 mL) and extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo to provide a residue. The residue was purified by preparative thin-layer chromatography eluting with dichloromethane to afford 7-methoxy-2- (p-tolyl)isoindolin-l-one 46 as an off-white solid (30 mg, 8%). LCMS analysis of 46: 98% and [M+H] ⁺ = 254.

Example 18. 2-(2-(trifluoromethyl)phenyl)isoindolin-l-one (48)

Preparation of 2-(2-(trifluoromethyl)phenyl)isoindolin-l-one (48)

To a round-bottomed flask were added methyl 2-(bromomethyl)benzoate 47 (228 mg, 1 mmol), 2-(trifluoromethyl)aniline 33 (161 mg, 1 mmol), acetonitrile (10 mL), and potassium carbonate (280 mg, 2 mmol) and the mixture was stirred at 65 °C for 2 hours. The reaction mixture was filtered and the filtrate was treated with sodium methoxide (35% w/w solution in methanol, 10 mL) and methanol (5 mL). The resulting mixture was stirred at 55 °C for 15 minutes, then diluted with water (50 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo to provide a residue. The residue was purified by preparative thin-layer chromatography eluting with 2: 1 petroleum ether/ethyl acetate to afford 2-(2-(trifluoromethyl)phenyl)isoindolin-l-one 48 as a white solid (20 mg, 7%). LCMS analysis of 48: 100% and [M+H] ⁺ = 278.

Example 19. l-(2-methoxyphenyl)-4-(p-tolyl)-lH-l, 2, 3-triazole (52)

51

Preparation of l-(2-methoxyphenyl)-4-(p-tolyl)-lH-l, 2, 3-triazole (52)

To a round-bottomed flask were added l-iodo-2-methoxybenzene 49 (1.2 g, 5 mmol), l-ethynyl-4-methylbenzene 50 (580 mg, 5 mmol), DMSO/water (5: 1 v/v, 30 mL), copper iodide (100 mg, 0.52 mmol), sodium ascorbate (100 mg, 0.5 mmol), (1S,2S)-N1,N2- dimethyl cyclohexane- 1,2-diamine 51 (107 mg, 0.75 mmol), sodium azide (350 mg, 5.38 mmol) and the mixture stirred at room temperature for 16 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were concentrated in vacuo to provide a residue. The residue was purified by preparative thin-layer chromatography eluting with 3: 1 petroleum ether/ethyl acetate to afford 4-(2-methoxyphenyl)-l-(p-tolyl)-lH- 1,2, 3 -triazole 52 as a white solid (265 mg, 20%).

LCMS analysis of 52: 99% and [M+H] ⁺ = 266.

Example 20. 4-(2-methoxyphenyl)-l-(p-tolyl)-lH-l, 2, 3-triazole (55)

51

Preparation of 4-(2-methoxyphenyl)-l-(p-tolyl)-lH-l, 2, 3-triazole (55)

To a round-bottomed flask were added l-iodo-4-methylbenzene 53 (1.1 g, 5 mmol), l-ethynyl-2-methoxybenzene 54 (660 mg, 5 mmol), DMSO/water (5: 1 v/v, 30 mL), copper iodide (95 mg, 0.5 mmol), sodium ascorbate (100 mg, 0.5 mmol), (1S,2S)-N1,N2- dimethyl cyclohexane- 1,2-diamine 51 (107 mg, 0.75 mmol), sodium azide (350 mg, 5.38 mmol) and the mixture stirred at room temperature for 16 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were concentrated in vacuo to provide a residue. The residue was purified by preparative thin-layer chromatography eluting with 3 : 1 petroleum ether/ethyl acetate to afford 4-(2-methoxyphenyl)-l-(p-tolyl)-lH- 1,2, 3 -triazole 55 as an off-white solid (250 mg, 19%). LCMS analysis of 55: 99% and [M+H] ⁺ = 266.

Example 21. 7-methoxy-2-(2-(trifluoromethyl)phenyl)isoindolin-l-one (57)

Preparation of 7-methoxyisoindolin-l-one (56)

To a solution of methyl 2-(bromomethyl)-6-methoxybenzoate 45 (500 mg, 2 mmol) was added ammonia in methanol (7N, 3 mL, 21 mmol) and tetrahydrofuran. The mixture was then stirred at room temperature for 16 hours. The mixture was concentrated in vacuo and purified by preparative thin-layer chromatography eluting with 20: 1

dichloromethane/methanol to afford 7-methoxyisoindolin-l-one 56 as a white solid (260 mg, 80%). LCMS analysis of 56: 100% and [M+H] ⁺ = 164.

Preparation of 7-methoxy-2-(2-(trifluoromethyl)phenyl)isoindolin-l-one (58)

To a sealed tube were added 7-methoxyisoindolin-l-one 56 (82 mg, 0.5 mmol), 1- iodo-2-(trifluoromethyl)benzene 57 (177 mg, 0.65 mmol), copper iodide (7 mg, 0.037 mmol), (lR,2R)-Nl,N2-dimethylcyclohexane-l, 2-diamine 51a (20 mg, 0.14 mmol), potassium carbonate (140 mg, 1 mmol), and toluene (4 mL). The tube was filled with an atmosphere of nitrogen and sealed. The mixture was stirred at 115 °C for 16 hours. The reaction mixture was cooled and concentrated in vacuo to provide a residue. The residue was purified by preparative thin-layer chromatography eluting with di chi orom ethane to afford 7-methoxy-2- (2-(trifluoromethyl)phenyl)isoindolin-l-one 58 as a white solid (15 mg, 10%). LCMS analysis of 58: 98% and [M+H] ⁺ = 308.

Example 22. 8-methoxy-2-(2-(trifluorom ethyl )phenyl)-3,4-dihydroisoquinolin-l(2H)-one (61)

Preparation of 8-methoxy-3,4-dihydroisoquinolin-l(2H)-one (60)

To a solution of 7-m ethoxy-2, 3 -dihydro- lH-inden-1 -one 59 (1.1 g, 6.8 mmol) in concentrated hydrochloric acid (20 mL) was added sodium azide (900 mg, 13.8 mmol) portionwise. The reaction mixture was stirred at room temperature for 16 hours, then diluted with ice and basified with sodium carbonate. The resulting mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo to provide a residue. The residue was purified by flash chromatography over silica gel eluting with 1 : 1 dichloromethane/ethyl acetate to afford 8-methoxy-3,4-dihydroisoquinolin-l(2H)-one 60 as a white solid (400 mg, 33%). LCMS analysis of 60: 82% and [M+H] ⁺ = 178.

Preparation of 8-methoxy-2-(2-(trifluoromethyl)phenyl)-3,4-dihydroisoquinol in-l(2H)-one (61)

To a sealed tube were added 8-methoxy-3,4-dihydroisoquinolin-l(2H)-one 60 (250 mg, 1.4 mmol), l-iodo-2-(trifluoromethyl)benzene 57 (500 mg, 1.8 mmol), copper iodide (25 mg, 0.14 mmol), (lS,2S)-Nl,N2-dimethylcyclohexane- 1,2-diamine 51 (80 mg, 0.56 mmol), potassium carbonate (483 mg, 3.5 mmol), and toluene (10 mL). The tube was filled with an atmosphere of nitrogen and sealed. The mixture was stirred at 140 °C for 28 hours. The reaction mixture was cooled, diluted with water (30 mL), and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated in vacuo to provide a residue. The residue was purified by preparative thin- layer chromatography eluting with 3:2 petroleum ether/ethyl acetate to afford 8-methoxy-2- (2-(trifluoromethyl)phenyl)-3,4-dihydroisoquinolin-l(2H)-one 61 as an off-white solid (120 mg, 26%). LCMS analysis of 61: 100% and [M+H] ⁺ = 322.

The compounds provided herein may be tested in assays and models described in the literature, including e.g., Peal, D. S. et al., Circulation, 2011 Jan 4; 123(1): 23-30. Examples of assays and models are also described below.

Example A. LQT2 Testing

The compounds provided herein can be tested for the ability to rescue zebrafish LQT2 breakdance embryos. The zebrafish breakdance (hkd) mutant carries an I59S mutation in KCNH2, the LQTS 2 gene, and has an easily observed cardiac phenotype. Moreover, the 159S KCNH2 mutation faithfully recapitulates in the zebrafish embryos many features of human LQTS: ventricular action potential durations (APD) prolongation, spontaneous early afterdepolarizations, and 2: 1 atrioventricular conduction (AV) block in early stages of development.

Breakdance homozygotes (bkd ^{~ ~} ) develop 2: 1 atrioventricular conduction block (two atrial beats for each ventricular beat) that is the result of a prolonged refractory period in the ventricle. This phenotype can be scored in vivo , owing to the transparency of the embryo, enabling a straightforward small molecule suppressor screen. Breakdance homozygotes are viable and fertile, with phenotypic penetrance of 92-96%.

Tubingen AB (TuAB) and tb218 (bkdt ¹ ) fish are maintained using standard methods. For screening, tb218 ^/_ adults are crossed, and embryos re reared in E3 buffer at 25°C. 24 hours post-fertilization (hpf) animals are dechorionated with proteinase-K. In order to minimize false positive results, three bkcd ¹ embryos can be plated in each well and compounds are scored as a hit if all three embryos are rescued. Accordingly, dechorionated embryos are plated three per well in 96-well plates in a final volume of 200 pL. Compounds are added to a final concentration of 10 ng/pL. Zebrafish are visually scored at 72 hpf for presence or absence of a 2: 1 AV block. The embryos re tested after the onset of

atrioventricular block in order to identify compounds that could specifically treat LQTS rather than compounds that exert their effects primarily by altering cardiac development.

Example B. Optical mapping

In order to directly measure the effects of the compounds provided herein on myocardial repolarization, ventricular action potential durations can be measured using voltage sensitive optical mapping in bkd ^~ and wildtype fish treated with active compounds or diluent. Optical voltage mapping of embryonic hearts can be performed as described previously (see Brunner, M. et al. J Clin Invest 118, 2246-59 (2008)). Mapping is performed on tb218 or TuAB hearts from embryos that has been treated with 100 mM of 2-MMB and flurandrenolide starting at 48 hpf. Explanted hearts are mapped at 72-75 hpf for 2-MMB treatment, and at 96-99 hpf for flurandrenolide treatment. Control fish for each time point is treated with 0.1% DMSO.

Example C. iPSC derived cardiomyocytes LQT3 and LQT1 assay

Stem-cell cultures were maintained in feeder-free culture in Essential 8 medium on Geltrex (Invitrogen) extracellular matrix. Differentiation into cardio yocytes was accomplished by combining an extracellular matrix sandwich using an overlay of dilute Geltrex along with temporal modulation of canonical Wnt signaling. Beating cardiomyocytes were observed in 10-20 days and micro-dissected from surrounding cells between 30-60 days and dissociated using 0.25% trypsin solution into small clusters onto Geltrex-coated coverslips. Dissociated cardiomyocytes were lentivirally transduced with the genetically-encoded voltage reporter, ArcLight (Leyton -Mange JS1, et al., Stem Cell Reports. 2014 Feb

6;2(2): 163-70), and imaged 72 hours later on a Nikon Eclipse Ti scope fitted with an environmental control chamber (In Vivo Scientific: CH.IVS.100) surrounding the stage to maintain humidity, temperature at 37°C, and supplied with 5% CO2 / 21% O2 / balance N2 in cardiomyocyte extracellular solution (Table SI). Coverslips were placed within an imaging chamber capable of field-stimulation (Warner Instruments: RC-49MFSH) and paced at lHz (unless otherwise noted) with a biphasic 8V for 20ms stimulus with at least 5 stimulations prior to image acquisition. Images were acquired with a FITC filter cube and an Evolve 128 (Photometries) camera at 1000 frames/sec. Spatially-averaged intensity traces were exported to in-house-developed analysis software (MATLAB) where traces were filtered and normalized (-AF/FR _es tmg) after which individual AP’s were evaluated for activation and 80% recovery to determine APD80. Cells were assessed immediately prior to drug treatment and again after 10 minutes of treatment.

iPSC recordings are taken from the same cell cluster at Baseline (>=10 min acclimation within the testing chamber), and after 5min of exposure to 0.5, 5, or 50uM of compound. WT vs KCNJ11 recordings performed 3months later using the same DMSO- dissolved aliquot of the compounds used in the iPSC recordings. Each compound was first tested on a WT cell cluster to assess continued efficacy of the formulation before testing on KCNJ11 KO cells. 2MMB was a positive control.

Example D. Results of LQT1, LQT2, and LQT3 Testing

Certain compounds were assessed in Zebrafish larvae for LQT2 (Example A) and in iPSC derived cardiomyocytes LQT3 and LQT1 assay (Example C).

The LQT3 iPSC cells did not have very prolonged action potentials, which likely reduces the sensitivity of this assay. The potentials are depicted as tracings with single recordings - solid line is baseline, and dotted line is treated.

LQT1 cells had action potentials that were long, as expected. These recordings were carried out in duplicate well on the multiwell plate and are shown with double tracings - solid line is baseline and dotted line is drug treated.

For the zebrafish LQT2 assay, the genetic mutants did not display 100% phenotype and a known IKr blocker (dofetilide) was added to achieve 100% penetrance at baseline. This may have reduced the sensitivity of this assay. Zebrafish results are shown as (number of fish rescued)/ (number of fish tested).

Three sets of control tracings are shown at the end of the table to illustrate the variability of the assay.

2MMB was a positive control. The second compound in Table 1 appears to have a larger absolute effect on APD than 2MMB. Compounds 6 and 7 were active in both assays. Compounds 20, 21, and 22 were active in the iPSC assay, but not the zebrafish assay.

Table 1

Various modifications of the invention, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference, including without limitation all patent, patent applications, and publications, cited in the present application is incorporated herein by reference in its entirety.