RU2838187C2 - Compounds - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula (I)

or a pharmaceutically acceptable salt thereof, where: R¹, R², R³ and R⁴ are independently selected from: (i) H; (ii) C_1-3 alkyl, optionally substituted with: hydroxy, C_1-2 alkoxy, NH₂, phenyl, C_5-6 heteroaryl, C_1-4 alkylcarbamoyl, or acylamido; (iii) C_1-3 alkoxy optionally substituted with C_3-6 cycloalkyl or one or more fluoro groups; (iv) C_3-6 cycloalkyl; (v) halogen; (vi) COR^C, where R^C is selected from NR^N1R^N2, where R^N1 and R^N2 are independently selected from H and methyl; (vii) cyano, NH₂ or NO₂; and (viii) phenyl or C_5-6 heteroaryl, optionally substituted with methyl, cyano, hydroxy or methoxy; Ar is phenyl optionally substituted with one or more groups selected from: (i) C_1-4 alkyl optionally substituted with hydroxy, C_1-2 alkoxy, NH₂, C_1-4 alkylcarbamoyl or one or more fluoro groups; (ii) C_3-6 cycloalkyl; (iii) hydroxy; cyano; NR^N3R^N4, where R^N3 and R^N4 are independently selected from H and methyl; or acylamido; (iv) halogen; (v) C_1-3 alkoxy, optionally substituted with hydroxy, C(O)NH₂, C_3-6 cycloalkyl, phenyl, C_5-6 heteroaryl or one or more fluoro groups; (vi) phenoxy, optionally substituted with fluorine; (vii) phenyl or C_5-6 heteroaryl; or (viii) SF₅ or SO₂CH₃; and, where at least one of R¹, R², R³ and R⁴ is not H and where C_5-6 heteroaryl has 5 to 6 ring atoms, of which 1 to 3 ring atoms are heteroatoms selected from a nitrogen atom, oxygen or sulphur, as well as a pharmaceutical composition which inhibits lysine acetyltransferase (KAT).

EFFECT: novel compounds which inhibit the activity of one or more KATs from the MYST family are obtained, and can be used for treating cancer.

14 cl, 17 tbl, 252 ex

Description

The present invention relates to compounds that act as inhibitors of lysine acetyltransferase (KAT) of the MYST family.

Prerequisites for the creation of an invention

The MYST family is the largest KAT family and is named after the yeast and mammalian representatives in which they were first discovered: MOZ, Ybf2/Sas3, Sas2, and TIP60 (Dekker 2014) . MYST proteins mediate many biological functions, including gene regulation, DNA repair, cell cycle regulation, and development (Avvakumov 2007; Voss 2009) . MYST family KAT proteins play a key role in post-translational modification of histones and thus have a profound impact on chromatin structure in the eukaryotic nucleus (Avvakumov 2007) . The family currently includes five mammalian KATs: TIP60 (KAT5; HTATIP; MIM 601409), MOZ (KAT6A; MIM 601408; MYST3), MORF (KAT6b; QKF; MYST4), HBO (KAT8; HBO1; MYST2), and MOF (KAT8; MYST1) (Voss 2009) . These five MYST family members are present in humans, and dysfunction of MYST proteins is known to be associated with cancer (Avvakumov 2007) . The most commonly used names for MYST family members are:

Common name Name MYST Systematic name MOF MYST1 KAT8 HBO MYST2 KAT7 MOZ MYST3 KAT6A MORF MYST4 KAT6B TIP60 KAT5

MYST Functional Domains

MYST proteins function in multi-subunit protein complexes including adaptors such as ING proteins that mediate DNA binding (Avvakumov 2007) . For example, TIP60 is associated with the NuA4 multi-protein complex (which includes more than 16 members) (Zhang 2017) . However, several reports have also been made of a helix-turn-helix DNA-binding motif within the MOZ protein structure itself (Holbert 2007) , suggesting the ability to bind directly to DNA.

The acetyltransferase activity of MYST proteins is influenced by the MYST domain (catalytic domain). The MYST domain contains an acetyl-CoA binding motif, which is structurally conserved with other HATs, and an unusual HC-type _C2 zinc finger (Voss 2009) . The highly conserved MYST domain, including the acetyl-CoA binding motif and zinc finger, is considered a defining feature of this enzyme family (Avvakumov 2007) .

The role of MYST proteins

Acetylation of histone residues is usually associated with transcriptional activation. However, in some cases, MYST proteins have also been implicated in transcriptional repression (Voss 2009) . Individual members of the MYST family are known to participate in a wide range of important biochemical interactions:

HBO1 activates DNA replication initiation (Avvakumov 2007; Aggarwal 2004; Doyon 2006; Iizuka 2006) through acetylation of histone substrates, which appears to result in a more accessible chromatin conformation (Avvakumov 2007, Iizuka 2006 ). HBO1 is also known to play a role in breast cancer pathogenesis by promoting the enrichment of cancer stem-like cells (Duong 2013) and destabilizing estrogen receptor α (ERα) through ubiquitination, which occurs through the histone acetylating activity of HBO1 (Iizuka 2013) . HBO1 is also involved in the development of acute myeloid leukemia (AML) ( Shi 2015 ).

TIP60 (KAT5) is the most studied member of the MYST family. TIP60 plays an important role not only in transcriptional regulation but also in the repair of DNA damage, especially DNA double-strand breaks (DSBs) (Gil 2017) . TIP60 can acetylate p53, ATM, and c-Myc. TIP60 and MOF specifically acetylate lysine 120 (K120) in p53 upon DNA damage (Avvakumov 2007) . TIP60 also plays an important role in regulatory T cell (Treg) biology. FOXP3 is a master regulator in Treg development and function, and acetylation of FOXP3 by TIP60 has been shown to be required for FOXP3 activity ( Li 2007 , Xiao 2014 ). Highlighting this, conditional deletion of TIP60 in mice results in a lethal scurvy-like autoimmune disease mimicking the phenotype observed in FOXP3 knockout mice ( Xiao 2014 ). In malignancy, Treg cells can promote tumor progression by suppressing adaptive antitumor immunity.

MOF (males absent on the first protein) was initially identified as one of the components of dosage compensation in Drosophila and was classified as a member of the MYST family based on functional studies and sequencing analysis (Su 2016) . The human ortholog shows significant similarity to Drosophila MOF; it contains an acetyl-CoA binding site, a chromodomain (which binds histones), and a _C2 HC-type zinc finger (Su 2016) . MOF is a key enzyme for histone H4K16 acetylation, and MOF-containing complexes are involved in various essential cellular functions and have been linked to malignancies (Su 2016). In addition to global reduction in histone acetylation, MOF depletion in mammalian cells can result in abnormal gene transcription, particularly by causing abnormal expression of certain tumor suppressor genes or oncogenes, suggesting a critical role for MOF in tumorigenesis (Su 2016). For example, MOF KAT activity has been shown to be required for the maintenance of MLL-AF9 leukemia and may be important for multiple AML subtypes ( Valerio 2017 ).

KAT6B (Querkopf) was first identified in a screen for mutations in genes regulating the balance between proliferation and differentiation during embryonic development (Thomas 2000) . Mice homozygous for a mutant allele of KAT6B have severe defects in cerebral cortex development resulting from a severe reduction in both proliferation and differentiation, particularly in the cortical progenitor population, during embryonic development. KAT6B is required for the maintenance of the neural stem cell population in adults and is part of a system regulating the differentiation of stem cells into neurons (Merson 2006) . KAT6B is also mutated in rare forms of leukemia (Vizmanos 2003) .

The MOZ locus ranks as the 12th most frequently amplified region among all types of malignancies ( Zack 2013 ). MOZ is located within the 8p11-p12 amplicon, which is found at a frequency of about 10-15% in various malignancies, especially breast and ovarian cancers ( Turner-Ivey 2014 ). MOZ was first identified as a fusion partner of CREB-binding protein (CBP) during a study of a specific chromosomal translocation in acute myeloid leukemia (AML) ( Avvakumov 2007; Borrow 1996 ) . MOZ KAT activity is required to stimulate the expression of MEIS1 and HOXa9 proteins, which are commonly overexpressed in some lymphomas and leukemias. Increased survival of MOZ ^+/- heterozygous mice has been observed in a transgenic model of E μ-Myc B-cell lymphoma, where loss of one MOZ allele results in a biologically significant reduction in Meis1 and Hoxa9 levels in B-cell precursors ( Sheikh 2015 ).

Inhibitors of some MYSTs are known. For example, the following anacardic acid derivative (Ghizzoni 2012) is reported to inhibit TIP60 (IC ₅₀ =74 μM) and MOF (IC ₅₀ =47 μM):

Other known inhibitors include (Zhang 2017):

In light of the established role of KAT in general and MYST in particular in diseases such as malignancies, there is a need for new inhibitors of these proteins.

Disclosure of invention

The present invention provides compounds that inhibit the activity of one or more KATs of the MYST family, i.e., TIP60, KAT6B, MOZ, HBO1 and MOF.

In a first aspect the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in a method of therapy:

(I)

Where:

R ¹ , R ² , R ³ and R ⁴ are independently selected from:

(i) H;

(ii) C _1-3 alkyl, optionally substituted by:

hydroxy,

C _1-2 alkoxy, optionally substituted with one or more fluoro groups,

_NH2 ,

phenyl,

C _5-6 heteroaryl,

C _1-4 alkylcarbamoyl,

acylamido or

one or more fluoro groups;

(iii) C _1-3 alkoxy optionally substituted with C _3-6 cycloalkyl or one or more fluoro groups;

(iv) C _3-6 cycloalkyl;

(v) halogen;

(vi) COR ^C , where R ^C is selected from NR ^N1 R ^N2 , where R ^N1 and R ^N2 are independently selected from H and methyl;

(vii) cyano, NH ₂ or NO ₂ ; and

(viii) phenyl or C _5-6 heteroaryl optionally substituted with methyl, cyano, hydroxy or methoxy;

Ar is a phenyl, naphthyl or C _5-10 heteroaryl group, which groups are optionally substituted with one or more groups selected from:

(i) C _1-4 alkyl optionally substituted with hydroxy, C _1-2 alkoxy, NH ₂ , C _1-4 alkylcarbamoyl or one or more fluoro groups;

(ii) C _3-6 cycloalkyl;

(iii) hydroxy; cyano; NR ^N3 R ^N4 , where R ^N3 and R ^N4 are independently selected from H and methyl; or acylamido;

(iv) halogen;

(v) C _1-3 alkoxy optionally substituted with hydroxy, C(O)NH ₂ , C _3-6 cycloalkyl, phenyl, C _5-6 heteroaryl or one or more fluoro groups;

(vi) phenoxy optionally substituted with fluorine;

(vii) phenyl or C _5-6 heteroaryl;

(viii) SF ₅ or SO ₂ CH ₃ ;

(ix) -(CH ₂ ) _n -Y-, where Y is O or CH ₂ and n is 2 or 3; or

(x) C _1-4 alkyl ester.

The first aspect also provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof according to the provided definition, and a pharmaceutically acceptable excipient.

In a second aspect of the present invention there is provided a method for treating an oncological disease, comprising administering to a patient in need of treatment a compound or a pharmaceutically acceptable salt thereof as defined in the first aspect of the invention, or a pharmaceutical composition according to the first aspect of the invention. In a second aspect of the present invention there is also provided the use of a compound or a pharmaceutically acceptable salt thereof as defined in the first aspect of the invention in the manufacture of a medicament for treating an oncological disease, and a compound or a pharmaceutically acceptable salt thereof as defined in the first aspect of the invention, or a pharmaceutical composition thereof, for use in treating an oncological disease.

As described below, in the treatment of an oncological disease, the compound according to the definition provided in the first aspect can be administered simultaneously or sequentially with radiation therapy and/or chemotherapy.

In a third aspect of the invention there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof:

(I)

Where:

R ¹ , R ² , R ³ and R ⁴ are independently selected from:

(i) H;

(ii) C _1-3 alkyl, optionally substituted by:

hydroxy,

C _1-2 alkoxy, optionally substituted with one or more fluoro groups,

_NH2 ,

phenyl,

C _5-6 heteroaryl,

C _1-4 alkylcarbamoyl,

acylamido or

one or more fluoro groups;

(iii) C _1-3 alkoxy optionally substituted with C _3-6 cycloalkyl or one or more fluoro groups;

(iv) C _3-6 cycloalkyl;

(v) halogen;

(vi) COR ^C , where R ^C is selected from NR ^N1 R ^N2 , where R ^N1 and R ^N2 are independently selected from H and methyl;

(vii) cyano, NH ₂ or NO ₂ ; and

(viii) phenyl or C _5-6 heteroaryl optionally substituted with methyl, cyano, hydroxy or methoxy;

Ar is a phenyl, naphthyl or C _5-10 heteroaryl group, which groups are optionally substituted with one or more groups selected from:

(i) C _1-4 alkyl optionally substituted with hydroxy, C _1-2 alkoxy, NH ₂ , C _1-4 alkylcarbamoyl or one or more fluoro groups;

(ii) C _3-6 cycloalkyl;

(iii) hydroxy; cyano; NR ^N3 R ^N4 , where R ^N3 and R ^N4 are independently selected from H and methyl; or acylamido;

(iv) halogen;

(v) C _1-3 alkoxy optionally substituted with hydroxy, C(O)NH ₂ , C _3-6 cycloalkyl, phenyl, C _5-6 heteroaryl or one or more fluoro groups;

(vi) phenoxy optionally substituted with fluorine;

(vii) phenyl or C _5-6 heteroaryl;

(viii) SF ₅ or SO ₂ CH ₃ ;

(ix) -(CH ₂ ) _n -Y-, where Y is O or CH ₂ and n is 2 or 3; or

(x) a C _1-4 alkyl ester;

provided that:

(a) at least one of R ¹ , R ² , R ³ and R ⁴ is not H, and optionally R ³ is not CF ₃ ; or

(b) R ⁴ is OMe; or

(c) R ⁴ is Cl, and either R ¹ , R ² and R ³ are H, or R ² is C1-3 alkyl or bromo, and R ¹ and R ³ are H; or

(d) R ³ is C1-3 alkyl, and R ¹ , R ² and R ⁴ are H.

In a fourth aspect of the present invention there is provided a synthesis of compounds according to the definition provided in the first or third aspects of the invention, as described below.

Definitions

Unless otherwise specified, the term "substituted" as used herein refers to a parent group that has one or more substituents. The term "substituent" is used herein in its generally accepted sense and refers to a chemical moiety that is covalently attached to or, where appropriate, fused to the parent group. The phrase "optionally substituted" as used herein refers to a parent group that may be unsubstituted or may be substituted.

C _5-12 heteroaryl: The term "C _5-12 heteroaryl" as used herein refers to a monovalent moiety obtained by removing a hydrogen atom from an aromatic structure having from 5 to 12 ring atoms, of which from 1 to 3 ring atoms are heteroatoms. The term "aromatic structure" is used to denote single ring or fused ring systems having aromatic properties, and the term "ring heteroatom" refers to a nitrogen, oxygen or sulfur atom.

In this context, prefixes (e.g. C _5-12 , C _5-6 , etc.) denote the number of atoms that make up the aromatic structure, or a range of the number of atoms that make up the aromatic structure, whether carbon atoms or heteroatoms.

Examples of C _5-12 heteroaryl structures include, but are not limited to, those derived from:

N ₁ : pyrrole (azole) (C ₅ ), pyridine (azine) (C ₆ ); pyridone (C ₆ ); indole (C ₉ ); quinoline (C ₁₀ );

O ₁ : furan (oxol) (C ₅ );

S ₁ : thiophene (thiol) (C ₅ );

N ₁ O ₁ : oxazole (C ₅ ), isoxazole (C ₅ ), isoxazine (C ₆ );

N ₂ O ₁ : oxadiazole (furazan) (C ₅ );

N ₁ S ₁ : thiazole (C ₅ ), isothiazole (C ₅ );

N ₂ S ₁ : thiadiazole (C ₅ )

N ₂ : imidazole (1,3-diazole) (C ₅ ), pyrazole (1,2-diazole) (C ₅ ), pyridazine (1,2-diazine) (C ₆ ), pyrimidine (1,3-diazine) (C ₆ ) (e.g. cytosine, thymine, uracil), pyrazine (1,4-diazine) (C ₆ ); benzimidazole (C ₉ )

N ₃ : triazole (C ₅ ), triazine (C ₆ ).

Halogen: The term "halogen" as used herein refers to a group selected from fluorine, chlorine, bromine and iodine.

Cyano: The term "cyano" as used herein refers to the -C≡N group.

Hydroxy: The term "hydroxyl" as used herein refers to the group -OH.

Phenyl: The term “phenyl” as used herein refers to a monovalent moiety obtained by removing a hydrogen atom from a single aromatic ring structure containing 6 carbon atoms in the ring (-C ₆ H ₅ ).

Phenoxy: The term “phenoxy” as used herein refers to the monovalent moiety obtained by removing a hydrogen atom from an oxygen atom in phenol (-OC ₆ H ₅ ).

C _1-4 alkyl: The term “C _1-4 alkyl” as used herein refers to a monovalent moiety obtained by removing a hydrogen atom from a carbon atom of a saturated hydrocarbon compound containing from 1 to 4 carbon atoms.

Examples of saturated alkyl groups include, without limitation, methyl (C ₁ ), ethyl (C ₂ ), propyl (C ₃ ), and butyl (C ₄ ).

Examples of saturated linear alkyl groups include, without limitation, methyl (C ₁ ), ethyl (C ₂ ), n-propyl (C ₃ ), and n-butyl (C ₄ ).

Examples of saturated branched alkyl groups include iso -propyl ( _C3 ), iso -butyl ( _C4 ), sec -butyl ( _C4 ), and tert -butyl ( _C4 ).

_C3-6 cycloalkyl: The term " _C3-6 cycloalkyl" as used herein refers to a monovalent moiety derived by removing a hydrogen atom from a carbon atom of a saturated cyclic hydrocarbon compound containing from 3 to 6 carbon atoms. Examples of _C3-6 cycloalkyl groups include, but are not limited to, cyclopropyl ( _C3 ), cyclobutyl ( _C4 ), cyclopentyl ( _C5 ), and cyclohexyl ( _C6 ).

C _1-4 alkoxy: The term "C _1-4 alkoxy" as used herein refers to a monovalent moiety derived by removing a hydrogen atom from an oxygen atom of a saturated alcohol compound containing from 1 to 4 carbon atoms. It can be represented as -OC _1-4 alkyl. Examples of C _1-4 alkoxy groups include, but are not limited to, methoxy (C ₁ ), ethoxy (C ₂ ), propyloxy (C ₃ ), and butyloxy (C ₄ ).

C _1-4 alkylcarbamoyl: -NHC(=O)OR, wherein R is a C _1-4 alkyl group as defined above. Examples of C _1-4 alkylcarbamoyl include, but are not limited to, -N(H)C(=O)OCH ₃ , -N(H)C(=O)OCH ₂ CH ₃ and -N(H)C(=O)OC(CH ₃ ) ₃ .

Acylamido: -NR(C=O)R', where R and R' are independently selected from H and C _1-4 alkyl as defined above. R and R' can also be -(CH ₂ ) _n -, where n is 3 or 4. Examples of an acylamido group include, but are not limited to, -N(H)C(=O)CF ₃ , N(H)C(=O)Me, and:

C _1-4 alkyl ester: The term "C _1-4 alkyl ester" as used herein refers to a monovalent moiety derived by removing a hydrogen atom from an oxygen atom of a saturated carboxylic acid compound having from 1 to 5 carbon atoms. It can be represented as -OC(O)-C _1-4 alkyl. Examples of C _1-4 alkyl ester groups include, but are not limited to, acetoxy (-OC(O)-CH ₃ ), propanoyloxy (-OC(O)-CH ₂ CH ₃ ), butanoyloxy (-OC(O)-CH ₂ CH ₂ CH ₃ ) and pentanoyloxy (-OC(O)-CH ₂ CH ₂ CH ₂ CH ₃ ).

Inclusion of other forms

Unless otherwise indicated, the above description includes the well-known ionic, salt, solvate and protected forms of these substituents. For example, reference to a carboxylic acid (-COOH) also includes the anionic (carboxylate) form (-COO ^- ), a salt or solvate thereof, as well as the usual protected forms. Similarly, reference to an amino group includes the protonated form (-N ⁺ HR ¹ R ² ), a salt or solvate of the amino group, for example, the hydrochloride salt, as well as the usual protected forms of the amino group. Similarly, reference to a hydroxyl group also includes the anionic form (-O ^- ), a salt or solvate thereof, as well as the usual protected forms.

Salts

It may be convenient or desirable to prepare, purify and/or use a corresponding salt of the active compound, for example a pharmaceutically acceptable salt. Examples of pharmaceutically acceptable salts are described in Berge 1977 .

For example, if the compound is anionic or has a functional group that can be anionic (e.g., -COOH can be ^-COO- ), then a salt can be formed with a suitable cation. Examples of suitable inorganic cations include, but are not limited to, alkali metal ions such as Na ⁺ and K ⁺ , alkaline earth metal cations such as Ca2 ⁺ and Mg2 ⁺ , and other cations such as Al ⁺³ . Examples of suitable organic cations include ^, but are not limited to, ammonium ion (i.e., _NH4 ⁺ ) and substituted ammonium ions (e.g., _NH3R ⁺ , _{NH2R2 +} , _NHR3 ⁺ , _NR4 ⁺ ). Examples of some suitable substituted ammonium ions are those derived from: ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, ^{phenylbenzylamine} , choline, meglumine, and tromethamine, as well as amino acids such as lysine and arginine. An example of a common quaternary ammonium ion is N( _CH3 ) ₄₊ .

If the compound is cationic or has a functional group that can be cationic (e.g. _-NH2 can be _-NH3 ⁺ ), then the salt can be formed with a suitable anion. Examples of suitable inorganic anions include, but are not limited to, anions derived from the following inorganic acids: hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric, nitrous, phosphoric, and phosphorous.

Examples of suitable organic anions include, but are not limited to, salts derived from the following organic acids: 2-acetyloxybenzoic, acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric, edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucoheptone, gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalenecarboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic, methanesulfonic, mucinous, oleic, oxalic, palmitic, pamoic, pantothenic, phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic, succinic, sulfanilic, tartaric, toluenesulfonic trifluoroacetic acid and valerianic acid. Examples of suitable polymeric organic anions include, but are not limited to, those derived from the following polymeric acids: tannic acid, carboxymethylcellulose.

Solvates

It may be convenient or desirable to prepare, purify and/or use a corresponding solvate of the active compound. The term "solvate" is used herein in the usual sense to refer to a complex of a solute (e.g., an active compound, a salt of the active compound) and a solvent. If the solvent is water, the solvate may conveniently be referred to as a hydrate, e.g., a monohydrate, a dihydrate, a trihydrate, etc.

Isomers

Certain compounds of the invention may exist in one or more specific geometric, optical, enantiomeric, diastereomeric, epimeric, atropic, stereoisomeric, tautomeric, conformational, or anomeric forms, including, without limitation, cis and trans forms; E and Z forms; c, t, and r forms; endo and exo forms; R, S, and meso forms; D and L forms; d and I forms; (+) and (-) forms; keto, enol, and enolate forms; syn and anti forms; synclinal and anticlinal forms; α and β forms; acacial and equatorial forms; bath, armchair, twist, envelope, and half-armchair forms; and combinations thereof, which are hereinafter referred to by the general term "isomers" (or "isomeric forms").

The term "chiral" refers to molecules that have the property of being non-superimposable on the mirror image of their partner, whereas the term "achiral" refers to molecules that are superimposable on the mirror image of their partner.

The term "stereoisomer" refers to compounds that have the same chemical composition but differ in the arrangement of atoms or groups in space.

"Diastereomer" refers to a stereoisomer with two or more centers of chirality, the molecules of which are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers can be separated by high-resolution analytical procedures such as electrophoresis and chromatography.

"Enantiomers" refers to two stereoisomers of a compound that are non-superimposable mirror images of each other.

Stereochemical definitions and notations used herein are generally those of S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., “Stereochemistry of Organic Compounds,” John Wiley & Sons, Inc., New York, 1994. The compounds of the invention may contain asymmetric or chiral centers and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the invention, including, without limitation, diastereomers, enantiomers, and atropisomers, as well as mixtures thereof, such as racemic mixtures, are part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. When describing an optically active compound, the prefixes D and L, or R and S , are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and I, or (+) and (-), are used to denote the sign of the direction of rotation of plane-polarized light by the compound, with (-) or I indicating that the compound is levorotatory. A compound with the prefix (+) or d is dextrorotatory. For the chemical structure shown, these stereoisomers are the same except that they are mirror images of each other. Also, a particular stereoisomer may be called an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, and it may occur in a chemical reaction or process where there is no stereoselectivity or stereospecificity. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomeric compounds without optical activity.

In the present invention, the carbon atom to which R ¹ and Cy are bonded may be a stereochemical center, i.e., when R ¹ is not H and R ¹ and Cy are different. The compounds of the present invention may be a racemic mixture or may be in enantiomeric excess or substantially enantiomerically pure.

It should be noted that, except as described below for tautomeric forms specifically excluded from the term "isomers" as used herein, these are structural (or constitutional) isomers (i.e., isomers that differ in the bonds between atoms and not simply in the position of the atoms in space). For example, reference to a methoxy group, _-OCH3, should not be construed as a reference to its structural isomer, the hydroxymethyl group, _-CH2OH . Similarly, reference to ortho-chlorophenyl should not be construed as a reference to its structural isomer, meta-chlorophenyl. However, reference to a class of structures may include structurally isomeric forms falling within that class (e.g., _C1-7 alkyl includes n-propyl and isopropyl; butyl includes n-, iso-, sec-, and tert-butyl; methoxyphenyl includes ortho-, meta-, and para-methoxyphenyl).

The above exception does not apply to tautomeric forms, such as keto forms, enol and enolate forms, such as in the following tautomeric pairs: keto/enol (shown below), imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime, thioketone/enthiol, N-nitroso/hydroxyazo and nitro/acin-nitro.

The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible when crossing a low-energy barrier. For example, proton tautomers (also known as prototropic tautomers) involve interconversions involving a proton transfer, such as keto-enol and imine-enamine isomerizations. Valence tautomers involve interconversions involving rearrangement of some of the bonding electrons.

It should be noted that the term "isomer" specifically includes compounds with one or more isotopic substitutions. For example, H may be in any isotopic form including ¹ H, ² H (D), and ³ H (T); C may be in any isotopic form including ¹² C, ¹³ C, and ¹⁴ C; O may be in any isotopic form including ¹⁶ O and ¹⁸ O; etc.

Examples of isotopes that may be included in the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as, but not limited to, ² H (deuterium, D), ³ H (tritium), ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ Cl, and ¹²⁵ I. Included are various isotopically labeled compounds of the present invention, such as those into which radioactive isotopes such as 3 H, 13 C, and 14 C are incorporated. Such isotopically labeled compounds may be useful in metabolic studies, in reaction kinetic studies, in detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT) involving drug or substrate tissue distribution assays, or in radiation therapy of patients. Therapeutic compounds of the invention labeled or substituted with deuterium may have improved DMPK (drug metabolism and pharmacokinetics) properties related to absorption, distribution, metabolism and excretion (ADME). Substitution with heavier isotopes such as deuterium may provide some therapeutic advantages due to greater metabolic stability of the compounds, such as a longer half-life in vivo or a reduced dosage requirement. A compound labeled with 18F may be useful for PET or SPECT studies. The isotopically labeled compounds of the present invention and their prodrugs can be prepared essentially by carrying out the procedures according to the schemes or examples and preparation methods described below, by replacing a reagent not containing isotopically labeled atoms with an available reagent with isotopically labeled atoms. In addition, substitution with heavier isotopes (i.e., 2H or D) can provide some therapeutic advantages due to greater metabolic stability of the compounds, such as a longer half-life in vivo, or a reduced dose requirement, or an improved therapeutic index. It is understood that deuterium is considered as a substituent in this context. The concentration of such a heavier isotope, in particular deuterium, can be determined by an isotopic enrichment factor. In the compounds of the present invention, any atom not specifically designated as a particular isotope means any stable isotope of that atom.

Unless otherwise indicated, reference to a particular compound includes all such isomeric forms, including (in whole or in part) racemic and other mixtures thereof. Methods for preparing (e.g., asymmetric synthesis) and separating (e.g., fractional crystallization and chromatographic methods) such isomeric forms are either known in the art or are readily accomplished by adapting in a known manner the methods described herein or known methods.

Inhibition

The compounds of the present invention inhibit the activity of one or more KATs of the MYST family, i.e., TIP60, KAT6B, MOZ, HBO1 and MOF.

The inhibitory activity of the compounds of the present invention is likely to vary depending on the KAT of the MYST family.

The compounds of the present invention may selectively inhibit the activity of one or more MYST family KATs compared to other MYST family KATs, i.e., the inhibitory activity of the compound may be higher for one or more MYST family KATs compared to one or more other MYST family KATs.

The compounds of the present invention can (selectively) inhibit the activity of one KAT of the MYST family. The compounds of the present invention can inhibit the activity of TIP60, MORF, MOZ, HBO1 or MOF.

The compounds of the present invention can inhibit the activity of two KATs of the MYST family, such as MOZ and MORF.

The compounds of the present invention can inhibit the activity of three KATs of the MYST family, such as MOZ, MORF and HBO1.

The compounds of the present invention can inhibit the activity of four KATs of the MYST family, such as MOZ, MORF, HBO1 and TIP60.

The compounds of the present invention can inhibit the activity of all five KATs of the MYST family, thus the compounds can inhibit the activity of TIP60, MORF, MOZ, HBO1 and MOF.

The compounds of the present invention may, in particular, inhibit the activity of MOZ and/or KAT6B and/or HBO1 and/or TIP60.

Indications for use

The compounds disclosed herein may provide therapeutic benefit in a number of disorders, particularly in the treatment or prevention of malignancies.

Oncological diseases

Inhibitors of post-translational lysine acetylation mediated by MYST family KATs are considered promising antitumor agents and may therefore be useful therapeutic agents, for example for use in the treatment of cancer. Such agents may also be useful as therapeutic agents for the treatment of malignancies that exhibit overexpression of MYST proteins.

"Oncological disease" may be any form of malignant neoplasm. In particular, oncological disease may include any one or more of the following: leukemia, acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), non-Hodgkin's lymphoma, Hodgkin's disease, prostate cancer, lung cancer, melanoma, breast cancer, colorectal cancer, colon cancer, squamous cell carcinoma, and gastric cancer.

Alternatively, the cancer may be adrenal cancer, anal cancer, bladder cancer, hematological malignancy, bone cancer, brain tumor, gynecologic cancer, gynecologic cancer, central nervous system lymphoma, cervical cancer, childhood-onset rhabdomyosarcoma, childhood-onset sarcoma, endometrial cancer, endometrial sarcoma, esophageal cancer, eye cancer, gallbladder cancer, gastrointestinal cancer, hairy cell leukemia, head and neck cancer, hepatocellular carcinoma, laryngeal cancer, Kaposi's sarcoma, renal cancer, laryngeal cancer, liver cancer, malignant fibrous histiocytoma, malignant thymoma, mesothelioma, multiple myeloma, myeloma, cancer of the nasal cavity and paranasal sinuses, cancer of the nasopharynx, cancer of the nervous system, neuroblastoma, oral cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary tumor, plasma cell neoplasm, primary CNS lymphoma, rectal cancer, respiratory tract cancer, retinoblastoma, salivary gland cancer, skin cancer, small bowel cancer, soft tissue sarcoma, gastric cancer, stomach cancer, testicular cancer, thyroid cancer, genitourinary sarcoma, uterine sarcoma, vaginal cancer, vascular cancer, Waldenstrom's macroglobulinemia and/or Wilms' tumor.

Malignant tumors can be classified into a specific type. Examples of types of malignant neoplasms include lymphoma, melanoma, carcinoma (eg, adenocarcinoma, hepatocellular carcinoma, medullary carcinoma, papillary carcinoma, squamous cell carcinoma), astrocytoma, glioma, medulloblastoma, myeloma, meningioma, neuroblastoma, sarcoma (eg, angiosarcoma, chondrosarcoma, osteosarcoma).

The malignant neoplasm may overexpress MYST. The malignant neoplasm may overexpress MYST protein compared to non-malignant tissue. In some cases, the malignant neoplasm produces MYST mRNA in excess compared to non-malignant tissue. The overexpressed MYST protein or MYST mRNA may be any one KAT from the MYST family, i.e., may be any of TIP60, KAT6B, MOZ, HBO1, and MOF. In some embodiments, the malignant neoplasm may overexpress more than one KAT from the MYST family, such as two or more selected from the group consisting of TIP60, KAT6B, MOZ, HBO1, and MOF. The malignant neoplasm may be one that evades immune recognition, such as through tumor-associated Treg cells.

Alternatively or additionally, the malignant neoplasm may overexpress a bromodomain: the malignant cell may overexpress one or more bromodomain-containing proteins (herein referred to as "bromodomain proteins") compared to non-malignant tissue. It may overproduce one or more bromodomain mRNAs compared to non-malignant tissue. In some cases, the level of bromodomain protein and/or mRNA in the cell is at a level approximately equivalent to the level in the non-malignant cell. The malignant neoplasm may overexpress one or more bromodomain proteins selected from the group consisting of: bromodomain protein (namely, BRD2, BRD3, BRD4, BRD7, BRD8, BRD9 and BRDT), TAF1/TAF1L, TFIID, SMARC2 (also referred to as BRM) and SMARC4 (also referred to as BRG1). For example, some colon cancers overexpress BRD8. Some acute myeloid leukemia cells overexpress BRD4.

Treg cells as a target for malignant neoplasms

Treg cells are immunosuppressive cells that prevent autoimmunity in the healthy mammalian immune system. However, some types of malignancies upregulate Treg activity to evade the host immune system. Treg infiltration in many tumor types correlates with poor prognosis for patients, and Treg depletion in tumor models demonstrates enhanced antitumor immune responses (Melero 2015) . Tumor-associated Treg suppression of the host immune system has been reported in lung (Joshi 2015), (Tso 2012) , breast (Gobert 2009; Yan 2011) , prostate (Miller 2006) , and pancreatic (Wang X 2016) cancers. FOXP3 is considered a master regulator of Treg cell differentiation, development, and function.

Several studies have demonstrated that FOXP3 acetylation plays a critical role in the stability of the FOXP3 protein and in regulating its ability to access DNA; and FOXP3 acetylation is mediated by KAT (Dhuban 2017) . Reducing TIP60-mediated FOXP3 acetylation has been shown to attenuate Treg development, suggesting an additional mechanism by which inhibition of the acetylating activity of MYST proteins may be exploited to intervene in diseases such as malignancies.

Combination therapy

The agents described herein may be used in combination with other anti-cancer therapies. They may act synergistically with chemo- or radiation therapy and/or with targeted therapeutic agents, including, but not limited to, FGFR1 inhibitors and therapeutic agents targeting nuclear hormone receptors. For example, the agents described herein may be used in combination with bromodomain-targeted drugs, including BET inhibitors. BET inhibitors reversibly bind to the bromodomains of the BET proteins: BRD2, BRD3, BRD4, and BRDT.

It is likely that inhibition of MYST family KAT proteins to reduce histone lysine acetylation (and other nuclear proteins described herein) will sensitize tumor cells to chemotherapy and radiotherapy by reducing DNA damage repair, such as double-strand break (DSB) repair, which increases chemotherapy- and radiotherapy-induced cancer cell death. Therefore, inhibition of MYST family KAT proteins is likely to synergize well with low-dose chemotherapy or radiotherapy.

Thus, in some cases, the MYST protein antagonist described herein may be administered in combination with a radiation therapy or chemotherapy regimen. It may be administered simultaneously or sequentially with radiation therapy and/or chemotherapy. Suitable chemotherapeutic agents and radiation therapy protocols will be readily apparent to one skilled in the art. In particular, the compound described herein may be combined with low-dose chemotherapy or radiation therapy. Suitable doses for "low-dose" chemotherapy or radiation therapy will be readily apparent to a skilled practitioner.

In particular, when the compounds of the present invention are used to eliminate Treg suppression, they can be combined with immune checkpoint inhibitors (Melero 2015, Wang L 2016 ). In addition, the compounds of the present invention that eliminate Treg suppression can be used in combination with radiation therapy to reduce Treg depletion in tumors ( Persa 2015, Jeong 2016 ).

Treatment methods

The compounds of the present invention can be used in a method of therapy. Also provided is a method of treatment comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of the invention . The term "therapeutically effective amount" means an amount sufficient to provide a benefit to the patient. Such benefit may consist of at least an improvement in at least one symptom. The actual amount administered, as well as the frequency and duration of administration, will depend on the nature and severity of the disease being treated. Prescribing treatment, such as deciding on a dose, is within the competence of physicians and other doctors.

As described above, the treatment of malignant neoplasms defined in this description may employ monotherapy or may, in addition to administering a compound of the invention, include standard surgical procedures, radiation therapy or chemotherapy. Such chemotherapy may include one or more of the following categories of antitumor agents:

(i) other antiproliferative/antineoplastic agents and combinations thereof used in medical oncology such as alkylating agents (e.g. cisplatin, oxaliplatin, carboplatin, cyclophosphamide, chlormethine, melphalan, chlorambucil, busulfan, temozolamide and nitrosoureas); antimetabolites (e.g. gemcitabine and antifolates such as fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside and hydroxyurea); antineoplastic antibiotics (e.g. anthracyclines such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin); antimitotic agents (eg, vinca alkaloids such as vincristine, vinblastine, vindesine, and vinorelbine, and taxoids such as taxol and docetaxel (taxotere), and polokinase inhibitors); and topoisomerase inhibitors (eg, epipodophyllotoxins such as etoposide and teniposide, amsacrine, topotecan, and camptothecin);

(ii) cytostatic agents such as antiestrogens (eg, tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and iodoxifene), antiandrogens (eg, bicalutamide, flutamide, nilutamide and cyproterone acetate), LHRH antagonists or LHRH agonists (eg, goserelin, leuprorelin and buserelin), progestogens (eg, megestrol acetate), aromatase inhibitors (eg, anastrozole, letrozole, vorazole and exemestane) and 5*-reductase inhibitors such as finasteride;

(iii) anti-invasion agents (e.g. c-Src kinase family inhibitors such as 4-(6-chloro-2,3-methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-tetrahydropyran-4-yloxyquinazoline (AZD0530; International Patent Application WO 01/94341), N-(2-chloro-6-methylphenyl)-2-{6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-ylamino}thiazole-5-carboxamide (dasatinib, BMS-354825; J. Med. Chem., 2004, 47, 6658–6661 and 4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-(3-(4-methylpiperazin-1-yl)propoxy)quinoline-3-carbonitrile (bosutinib, SKI-606; Cancer research (2003), 63(2), 375-81) and metalloproteinase inhibitors such as marimastat, urokinase plasminogen activator receptor function inhibitors, or heparanase antibodies);

(iv) inhibitors of growth factor function; for example, such inhibitors include antibodies to growth factors or growth factor receptors (e.g., the anti-erbB2 antibody tastuzumab [Herceptin T], the anti-EGFR antibody panitumumab, the anti-erbB1 antibody cetuximab [Erbitkus, C225], and any of the antibodies to growth factors or growth factor receptors disclosed in Stern 2005 ; such inhibitors also include tyrosine kinase inhibitors, such as inhibitors of the epidermal growth factor family (e.g., EGFR family tyrosine kinase inhibitors such as N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, ZD1839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774) and 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)quinazolin-4-amine (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib; hepatocyte growth factor family inhibitors; platelet-derived growth factor family inhibitors such as imatinib; serine/threonine kinase inhibitors (eg, Ras/Raf signaling inhibitors such as farnesyltransferase inhibitors, eg, sorafenib (BAY 43-9006)), inhibitors of cell signaling through MEK and/or AKT kinase, hepatocyte growth factor family inhibitors, c-kit inhibitors, abl kinase inhibitors, IGF-receptor (insulin-like growth factor) kinase inhibitors; aurora kinase inhibitors (eg, AZD1152, PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528, and AX39459) and cyclin-dependent kinase inhibitors such as CDK2 and/or CDK4 inhibitors;

(v) antiangiogenic and antilymphangiogenic agents, such as those that inhibit the action of vascular endothelial growth factor [e.g., the anti-vascular endothelial cell growth factor A (VEGFA) antibody bevacizumab (Avastin T), the anti-vascular endothelial cell growth factor A (VEGFA) antibody ranibizumab, the anti-VEGF aptamer pegaptanib, the anti-vascular endothelial growth factor receptor 3 (VEGFR3) antibody IMC-3C5, the anti-vascular endothelial cell growth factor C (VEGFC) antibody VGX-100, the anti-vascular endothelial cell growth factor D (VEGFD) antibody VGX-200, the soluble form of vascular endothelial growth factor receptor 3 (VEGFR3) VGX-300, and VEGF receptor tyrosine kinase inhibitors such as 4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline (vandetanib; ZD6474; Example 2 of WO 01/32651), 4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1-ylpropoxy)quinazoline (sediranib; AZD2171; Example 240 of WO 00/47212), vatalanib (PTK787; WO 98/35985), pazopanib (GW786034), axitinib (AG013736), sorafenib and sunitinib (SU11248; WO 01/60814), compounds such as those disclosed in international patent applications WO97/22596, WO 97/30035, WO 97/32856 and WO 98/13354 and compounds with other mechanisms of action (e.g. linomid, inhibitors of integrin function avb3 and angiostatin)];

(vi) vascular damaging agents such as combretastatin A4 and the compounds disclosed in International Patent Applications WO 99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and WO 02/08213;

(vii) antisense therapeutics, such as those directed at the targets listed above, such as ISIS 2503, an antisense anti-ras;

(viii) gene therapy approaches, including examples of approaches to replace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2, GDEPT (gene-directed enzyme drug therapy) approaches such as using cytosine deaminase, thymidine kinase or bacterial nitroreductase enzyme, and approaches to increase patient tolerance to chemotherapy or radiotherapy such as multidrug resistance gene therapy; and

(ix) immunotherapeutic approaches, including, for example, ex vivo and in vivo approaches to enhance the immunogenicity of a patient's tumor cells, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colony-stimulating factor, approaches to reduce T-cell anergy, approaches using transfected immune cells such as cytokine-transfected dendritic cells, approaches using cytokine-transfected tumor cell lines and approaches using anti-idiotypic antibodies

Introduction

The active compound or a pharmaceutical composition comprising the active compound can be administered to a subject by any convenient route of administration, systemically/peripherally or at the site of desired action, including, but not limited to, the following routes: oral (e.g., ingestion); topical (including, e.g., transdermal, intranasal, ocular, buccal, and sublingual); pulmonary (e.g., by inhalation or insufflation therapy using, e.g., an aerosol, e.g., through the mouth or nose); rectal; vaginal; parenteral, e.g., by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intra-articular, subarachnoid, intravitreal, and intrasternal injection; by implantation of a depot, such as subcutaneously, intravitreally or intramuscularly. The subject may be a eukaryote, an animal, a vertebrate, a mammal, a rodent (e.g., a guinea pig, a hamster, a rat, a mouse), a murine (e.g., a mouse), a canine (e.g., a dog), a feline (e.g., a cat), an equine (e.g., a horse), a primate, a monkey (e.g., a marmoset or ape), a monkey (e.g., a baboon), an ape (e.g., a gorilla, a chimpanzee, an orangutan, a gibbon) or a human.

Preparations

Although it is entirely possible for the active compound to be administered alone, it is desirably presented as a pharmaceutical composition (e.g., a preparation) containing at least one active compound as described above, together with one or more other pharmaceutically acceptable carriers, adjuvants, excipients, diluents, fillers, buffers, stabilizers, preservatives, lubricants or other materials well known to those skilled in the art, and optionally other therapeutic or prophylactic agents.

Thus, the present invention further relates to pharmaceutical compositions as defined above and methods for preparing pharmaceutical compositions comprising a mixture of at least one active compound as described above together with one or more other pharmaceutically acceptable carriers, excipients, buffers, adjuvants, stabilizers or other materials as described herein.

The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions and/or dosage forms that are, within the scope of medical judgment, suitable for use in contact with the tissues of a subject (e.g., a human) without undue toxicity, irritation, allergic reaction or other problems or complications, with a reasonable benefit/risk ratio. Each carrier, excipient, etc. must also be "acceptable" in the sense of being compatible with the other ingredients of the preparation.

Descriptions of suitable carriers, excipients, etc. can be found in standard pharmaceutical texts, such as Remington's Pharmaceutical Sciences , 18th edition, Mack Publishing Company, Easton, Pa., 1990.

The preparations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Such methods include the step of bringing the active compound into association with a carrier which constitutes one or more accessory ingredients. The preparations are generally prepared by uniformly and intimately mixing the active compound with liquid carriers or finely divided solid carriers or both, and then shaping the product if necessary.

Medicines may be presented as liquids, solutions, suspensions, emulsions, elixirs, syrups, tablets, lozenges, granules, powders, capsules, wafers, pills, ampoules, suppositories, pessaries, ointments, gels, pastes, creams, sprays, mists, foams, lotions, oils, boluses, medicated slurries or aerosols.

Formulations suitable for oral administration (e.g., by mouth) may be presented as discrete units such as capsules, cachets, or tablets, each containing a predetermined amount of the active compound; as a powder or granules; as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion, as a bolus, stool, or paste.

Tablets may be prepared by conventional means, for example, by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be made by compressing in a suitable machine the active compound in a free-flowing form such as a powder or granules, optionally mixed with one or more binders (e.g. povidone, gelatin, acacia, sorbitol, tragacanth, hydroxypropyl methylcellulose); fillers or diluents (e.g. lactose, microcrystalline cellulose, calcium hydrogen phosphate); lubricants (e.g. magnesium stearate, talc, silicon dioxide); disintegrating agents (e.g. sodium starch glycollate, cross-linked povidone, cross-linked sodium carboxymethylcellulose); surface active or dispersing or wetting agents (e.g. sodium lauryl sulfate); and preservatives (e.g., methyl para-hydroxybenzoate, propyl para-hydroxybenzoate, sorbic acid). Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be film coated or scored, or may be formulated to provide slow or controlled release of the active compound therefrom using, for example, hydroxypropyl methylcellulose in varying proportions to achieve the desired release profile. The tablets may optionally be enteric coated to provide release in areas of the digestive tract other than the stomach.

Formulations suitable for topical administration (e.g. transdermal, intranasal, ocular, buccal and sublingual) may be formulated as an ointment, cream, suspension, lotion, powder, solution, paste, gel, spray, aerosol or oil. Alternatively, the formulation may be a patch or dressing, such as a bandage or adhesive plaster, impregnated with the active compound and, optionally, one or more excipients or diluents.

Formulations suitable for topical administration in the mouth include lozenges containing the active compound in a flavoured base, usually sucrose and acacia or tragacanth; pastilles containing the active compound in an inert base such as gelatin and glycerol or sucrose and acacia; and mouthwashes containing the active compound in a suitable liquid carrier.

Formulations suitable for topical ocular administration include eye drops in which the active compound is dissolved or suspended in a suitable vehicle, especially an aqueous vehicle for the active compound.

Formulations suitable for nasal administration in which the carrier is a solid include coarse powders having a particle size of, for example, in the range of about 20 to about 500 microns, administered by inhalation, for example by rapid inhalation through the nasal passage, from a container of powder by holding it close to the nose. Suitable formulations in which the carrier is a liquid for administration, for example in the form of a nasal spray, nasal drops or a means for aerosol administration using a nebulizer, include aqueous or oily solutions of the active compound.

Preparations suitable for administration by inhalation include those presented in the form of an aerosol spray in pressurized packaging using a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases.

Formulations suitable for topical application to the skin include ointments, creams and emulsions. In preparing an ointment, the active compound is formulated, optionally using a paraffinic or water-miscible ointment base. Alternatively, the active compounds can be formulated into a cream using an oil-in-water cream base. If desired, the aqueous phase of the cream base can include, for example, at least about 30% by weight of a polyhydric alcohol, i.e., an alcohol containing two or more hydroxyl groups, such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol and mixtures thereof. Formulations for topical application can, if desired, include a compound that enhances the absorption or permeation of the active compound through the skin or other areas of action. Examples of such skin permeation enhancers include dimethyl sulfoxide and related analogs.

When incorporated into an emulsion for topical use, the oil phase may optionally contain only an emulsifier (otherwise known as an emulsifier), or may contain a mixture of at least one emulsifier with a fat or oil or both a fat and an oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier, which acts as a stabilizer. Also preferred is the inclusion of both an oil and a fat. The emulsifier(s), together with the stabilizer(s) or without the stabilizer(s), constitute the so-called emulsifying wax, and the wax together with the oil and/or fat form the so-called emulsifying ointment base, which creates the oily dispersion phase of the cream preparation.

Suitable emulsifiers and emulsion stabilizers include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulfate. The choice of suitable oils or fats for the formulation is based on achieving the desired cosmetic properties, since the solubility of the active compound in most oils likely to be used in pharmaceutical emulsion formulations may be very low. Thus, the cream should preferably be a non-greasy, non-staining and rinseable formulation with a suitable consistency to avoid leakage from tubes or other containers. Mono- or dibasic straight or branched chain alkyl esters such as diisoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a mixture of branched chain esters known as Crodamol CAP may be used, the last three being the preferred esters. They may be used singly or in combination, depending on the properties required.

Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin, or other mineral oils may be used.

Preparations suitable for rectal administration may be presented as a suppository with a suitable base containing, for example, cocoa butter or a salicylate.

Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams and aerosols containing in addition to the active compound such carriers as are known in the art to be appropriate.

Formulations suitable for parenteral administration (e.g., by injection, including cutaneous, subcutaneous, intramuscular, intravenous, and intradermal) include aqueous or nonaqueous, isotonic, pyrogen-free, sterile injection solutions which may contain antioxidants, buffers, preservatives, stabilizers, bacteriostats, and solutes which render the preparation isotonic with the blood of the intended recipient; and aqueous and nonaqueous sterile suspensions which may include suspending agents and thickening agents, as well as liposomes or other microparticulate systems which are intended to target the compound to blood components or one or more organs. Examples of suitable isotonic vehicles for use in such preparations include sodium chloride injection, Ringer's solution, or lactated Ringer's solution for injection. Typically, the concentration of the active compound in the solution is from about 1 ng/mL to about 10 μg/mL, such as from about 10 ng/mL to about 1 μg/mL. The formulations may be presented in closed single-dose or multi-dose containers, such as ampoules and vials, and may be stored in a sublimated (lyophilized) state requiring only the addition of a sterile liquid carrier, such as water for injection, immediately before use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets. The formulations may be in the form of liposomes or other microparticulate systems that are designed to target the active compound to blood components or one or more organs.

Dosage

It will be appreciated by those skilled in the art that appropriate dosages of the compound and compositions containing the compound may vary among patients. Determination of the optimal dosage typically involves balancing the level of therapeutic benefit against any risk or undesirable side effects. The dosage level selected will depend on a variety of factors including, without limitation, the activity of the particular compound, the route of administration, the time of administration, the rate of elimination of the compound, the duration of treatment, other drugs, compounds, and/or materials used in combination, the severity of the condition, and the species, sex, age, body weight, pathological condition, general health, and previous medical history of the patient. The amount of compound and the route of administration are ultimately determined by the judgment of the physician, veterinarian, or clinician, although in general the dosage is selected to provide local concentrations at the site of action that provide the desired effect without causing significant harmful or dangerous side effects.

Administration may be in a single dose, continuously, or intermittently (e.g., in divided doses at appropriate intervals) throughout the course of treatment. Methods for determining the most effective agents and doses to administer are well known to those skilled in the art and will vary depending on the agents being used for therapy, the purpose of therapy, the target cell(s) of treatment, and the subject being treated. Single or multiple administrations may be administered using a dose level and administration schedule selected by the attending physician, veterinarian, or clinician.

In general, a suitable dose of the active compound is in the range of about 100 ng to about 25 mg (more typically: about 1 μg to about 10 mg) per kilogram of body weight of the subject per day. In the case where the active compound is a salt, ester, amide, prodrug, etc., the amount administered is calculated on the basis of the parent compound so as to proportionally increase the actual weight applied.

In one embodiment, the active compound is administered to a human patient according to the following dosage regimen: approximately 100 mg 3 times daily.

In one embodiment, the active compound is administered to a human patient according to the following dosage regimen: approximately 150 mg twice daily.

In one embodiment, the active compound is administered to a human patient according to the following dosage regimen: approximately 200 mg twice daily.

However, in one embodiment, the active compound is administered to a human patient according to the following dosage regimen: about 50 or about 75 mg 3 or 4 times daily.

In one embodiment, the active compound is administered to a human patient according to the following dosage regimen: about 100 or about 125 mg twice daily.

Treatment

The term "treatment" as used herein in the context of treating a condition refers generally to treatment and therapy of a human or animal (e.g., veterinary use) that achieves some desired therapeutic effect, such as inhibition of the progression of a condition, and includes decreasing the rate of progression, inhibiting the rate of progression, regressing the condition, alleviating the condition, and curing the condition. Also included is treatment as a prophylactic measure (i.e., prevention, avoidance).

The term "therapeutically effective amount" as used herein refers to an amount of an active compound or a material, composition or dosage form containing the active compound that is effective to achieve some desired therapeutic effect, associated with a reasonable benefit/risk ratio when administered in accordance with the intended treatment regimen.

Similarly, the term "prophylactically effective amount" as used herein refers to an amount of an active compound or a material, composition or dosage form containing the active compound that is effective to achieve some desired prophylactic effect, associated with a reasonable benefit/risk ratio when administered in accordance with the intended treatment regimen.

Subject/Patient

The subject/patient may be an animal, a mammal, a placental mammal, a marsupial (e.g., kangaroo, wombat), a monoaperture (e.g., platypus), a rodent (e.g., guinea pig, hamster, rat, mouse), a murine (e.g., mouse), a lagomorph (e.g., rabbit), an avian (e.g., bird), a canine (e.g., dog), a feline (e.g., cat), an equine (e.g., horse), a porcine (e.g., pig), an ovine (e.g., sheep), a ruminant (e.g., cow), a primate, a monkey (e.g., marmoset or ape), a monkey (e.g., marmoset, baboon), ape (e.g., gorilla, chimpanzee, orangutan, gibbon), or a human.

In addition, the subject/patient may be at any stage of development, such as a fetus. In one preferred embodiment, the subject/patient is a human.

General methods of synthesis

The compounds of the invention can be prepared using the following general methods and procedures, which are described in detail for the examples. The reaction conditions mentioned are illustrative and not limiting, for example, a person skilled in the art can use a wide range of synthetic methods to synthesize the desired compounds, such as, but not limited to, methods described in the literature (e.g., but not limited to, March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 7th edition or Larock's Comprehensive Organic Transformations: Comprehensive Organic Transformations: A Guide to Functional Group Preparations).

The compounds of formula (l) described above can be prepared using the synthetic strategies outlined below, to which the above definitions apply.

General Synthesis 1

Scheme 1A illustrates the formation of a sulfonamide linkage to form compounds of structure I by coupling the corresponding sulfonyl chloride compound of structure G2 with a primary or secondary amine such as benzisoxazolamine G3 .

Scheme 1A

Methods for forming such sulfonamides will be obvious to those skilled in the art, but include, for example, the use of a suitable base such as, but not limited to, pyridine, LiHMDS, n -BuLi, or NaH, and the use of activated forms of the sulfonic acid such as the corresponding sulfonyl halide. The formation of sulfonyl chlorides of structure G2 from the corresponding acids of structure G1 can be achieved, for example, using thionyl chloride or cyanuric acid chloride.

Alternatively, an activated form of the sulfonic acid, such as, but not limited to, a pentafluorophenyl sulfonate ester or a trichlorophenyl sulfonate ester of structure G5 , can be coupled to an appropriate primary or secondary amine, such as a benzisoxazolamine G3 (Scheme 1B).

Scheme 1B

Formation of the sulfonate ester of form G5 from the corresponding sulfonyl chloride G2 and the corresponding phenol ( ^R5 may be, for example, pentafluorophenyl or trichlorophenyl) can be achieved using a suitable base, such as, but not limited to, pyridine or triethylamine. Methods for forming the sulfonamides in I will be apparent to those skilled in the art, but include, for example, using a suitable base, such as, but not limited to, LiHMDS.

General Synthesis 2

Scheme 2A illustrates the formation of a sulfonyl chloride, such as G2 , as a substituent that is part of Ar.

Scheme 2A

This can be achieved by reacting an appropriate aryl compound of structure G6 , such as, but not limited to, with chlorosulfonic acid. Alternatively, the aryl compound G6 can be treated sequentially with a base, such as, but not limited to, nBuLi , and sulfur dioxide to produce a lithium arylsulfinate, which is further oxidized, such as with sulfuryl chloride, to produce the desired sulfonyl chloride of structure G2 . The product G2 can be isolated by methods known to those skilled in the art, or can be prepared in situ and used immediately in a subsequent step.

Additionally, the sulfonyl chloride in structure G2 can be formed from the arylthiol in structure G8 shown in Scheme 2B.

Scheme 2B

Methods for forming G2 include, for example, using a suitable oxidizing agent such as, but not limited to, hydrogen peroxide and potassium nitrate in the presence of a chloride source such as, but not limited to, chlorotrimethylsilane or thionyl chloride. The thiol structure G8 can be synthesized from a compound of structure G7 , where (X) can be a halogen, by methods known to those skilled in the art, including, but not limited to, nucleophilic substitution in the presence or absence of a transition metal.

Alternatively, sulfonation of an aryl compound such as G6 can provide the corresponding sulfonic acid of structure G1 . This can be achieved using any suitable reagent known to those skilled in the art, such as sulfur trioxide or sulfuric acid.

Scheme 2C

Sulfonic acid G1 can be converted to sulfonyl chloride G2 by the routes shown in Scheme 1A of general synthesis 1.

General Synthesis 3

Scheme 3A illustrates the formation of a benzisoxazolamine, such as G3 , from an aryl nitrile with an ortho substituent X, such as structure G9 . The group (X) can be, without limitation, a halogen, such as a chlorine or fluorine group, and is selected to be suitable for the reaction employed.

Scheme 3A

For example, starting material G9 can be reacted with an oxime, such as, but not limited to, acetone oxime or, for example, acetohydroxamic acid, in the presence of a suitable base, such as, but not limited to, potassium tert -butoxide, to form benzisoxazolamine G3 .

General Synthesis 4

Scheme 4A

Scheme 4A illustrates the addition of an R ⁷ group to a compound of structure G10 (where R ⁶ is H or a suitable protecting group, including, but not limited to, 2,4-dimethoxybenzyl (DMB); methods for removing said protecting groups are known to those skilled in the art (e.g., Greene's Protective Groups in Organic Synthesis, 4th edition )), as a substituent that is part of Ar. This can be achieved using any suitable coupling reaction known to those skilled in the art, such as a Suzuki coupling. The R ⁷ B ¹ and X groups are selected to be suitable for the coupling reaction employed. For example, in the case of a Suzuki coupling, (X) can be a halogen, triflate, or other suitable group, and B ¹ is a suitable boron compound, including, but not limited to, a boronic acid or a boronate ester.

Examples of B ¹ that can be used in the Suzuki coupling reaction include, but are not limited to, those shown below.

The types of R ⁷ B ¹ compounds that can be used in the Suzuki coupling reaction include, but are not limited to, those shown below.

In addition to Scheme 4A, the position of (X) and (B ¹ ) can be reversed, as shown below in Scheme 4B, to yield the same final compound G11 . Similar to Scheme 2A, the groups designated R ⁷ X and B ¹ are chosen to be suitable for the coupling reaction being used. For example, in the case of a Suzuki coupling reaction, (X) can be a halogen, triflate, or other suitable group, and B ¹ is a suitable boron compound, including, but not limited to, a boronic acid or a boronate ester.

Scheme 4B

For the introduction of the R ⁷ group, various coupling reactions other than the Suzuki coupling can be used, such as, for example, coupling reactions catalyzed by transition metals or, for example, by compounds of tin (Stille reaction) and zinc (Negishi reaction). Furthermore, when, for example, the group (X) is, without limitation, phenol (OH) or a primary or secondary amine (R'R''NH), a Chan-Lam type coupling can be used.

The transformations shown in Schemes 4A and 4B can also be accomplished with the ^R1 , ^R2 , ^R3 , or ^R4 substituent in the benzisoxazole moiety in structure G13 shown in Scheme 4C below.

Scheme 4C

Additionally, the R ⁷ substituent can be introduced prior to the formation of the sulfonamide and benzisoxazole on the nitrile precursor G9 (where R ¹ , or R ² , or R ³ , or R ⁴ = X or B ¹ ), as shown in Scheme 3D of the general synthesis 3.

General Synthesis 5

Scheme 5A illustrates the addition of a nitrogen-linked ^R8 group as a substituent that is part of Ar or on a benzisoxazole moiety to yield a compound of structure G16 . This can be achieved using any suitable coupling reaction known to one skilled in the art, such as by displacement of SnAr or by a Buchwald coupling reaction. The group designated (X) can be, without limitation, a halogen and is selected to be suitable for the coupling reaction employed.

Scheme 5A

Alternatively, a similar strategy can be used to synthesize ether-linked compounds as shown in Scheme 5B. This can be achieved using any suitable coupling reaction known to those skilled in the art, such as by SnAr displacement or by a Ullmann coupling reaction, to give compounds with structure G17.

Scheme 5B

Both of the above bonds can also be reversed so that the added group is R ⁸ -X.

Additionally, an OR ⁸ or NHR ⁸ substituent can be introduced prior to the formation of the sulfonamide and benzisoxazole on the nitrile precursor G9 (where R ¹ , or R ² , or R ³ , or R ⁴ = X), as shown in Scheme 3D of the general synthesis 3.

General Synthesis 6

Scheme 6A

Scheme 6A illustrates the addition of an amine (HNR ¹⁰ R ¹¹ ) to form the corresponding amide as a substituent that is part of Ar or on a benzisoxazole moiety to yield a compound of structure G19 . This can be achieved by coupling the appropriate carboxylic acid with a primary amine or a secondary amine NHR ¹⁰ R ¹¹ . Methods for forming such amides will be apparent to those skilled in the art but include, for example, the use of reagents such as HATU, HBTU, T3P, and EDCI/HOBt, and the use of activated forms of the carboxylic acid such as the corresponding acyl halide, mixed anhydride, or N- hydroxysuccinimide ester. The amide G19 can also be synthesized directly from the ester compound (when R ¹² =Alk, such as, but not limited to, methyl or ethyl). The formation of a carboxylic acid (when R ¹² =H) from the corresponding ester can be achieved, for example, by hydrolysis with a base such as an alkali metal hydroxide or an acid such as aqueous hydrochloric acid.

Further transformations can be carried out from the amide compound G19 , such as, but not limited to, reduction of the amide to form an amine or dehydration of the amide to form a nitrile. Methods for carrying out such a transformation will be known to those skilled in the art.

In addition, the amide CONR ¹⁰ R ¹¹ can be introduced prior to the formation of the sulfonamide and benzisoxazole on the nitrile precursor G9 (where R ¹ , or R ² , or R ³ , or R ⁴ =CO ₂ R ¹² , R ¹² =Alk or H), as shown in Scheme 3D of the general synthesis 3.

General Synthesis 7

The conversion of (X) in structure G20 in Scheme 7A to the ester in structure G18 (R ¹² = alkyl such as methyl or ethyl) will be apparent to those skilled in the art, but involves, for example, a carbonylation reaction, which can be achieved using carbon monoxide in the presence of a transition metal catalyst such as, but not limited to, PdCl ₂ dppf⋅DCM; and an alcoholic solvent such as, but not limited to, methanol or ethanol. Formation of the carboxylic acid in structure G18 (R = H) can be achieved, for example, by hydrolysis with a base such as an alkali metal hydroxide or an acid such as aqueous hydrochloric acid.

Scheme 7A

The ester or acid G18 in Scheme 7A can be reduced to a hydroxyl compound such as structure G21 . Methods for such a transformation are known to those skilled in the art but include, for example, the use of reducing agents such as lithium aluminum hydride (for the ester and carboxylic acid) and borohydride (for the carboxylic acid).

Further transformations such as, but not limited to, Mitsunobu or nucleophilic substitution reactions can be carried out from the hydroxyl compound G21 . Methods for carrying out such a transformation will be known to those skilled in the art.

Additionally, an ester group can be introduced prior to the formation of the sulfonamide and benzisoxazole on the nitrile precursor G9 (where R ¹ , or R ² , or R ³ , or R ⁴ = X), as shown in Scheme 3D of the general synthesis 3.

General Synthesis 8

Scheme 8A illustrates the reduction of the nitro group in structure G22 to form the corresponding amine in structure G23 as a substituent that is part of Ar, or on the benzisoxazole moiety.

Scheme 8A

Reduction of the nitro group to the primary amine G23 will be apparent to those skilled in the art and includes, without limitation, the use of such reducing conditions as the use of a transition metal (Fe, In, Zn) in the presence of HCl, hydrogenation in the presence of a transition metal, or a transition metal catalyst.

Further transformations such as, but not limited to, the formation of an amide bond can be carried out from the amine compound G23 . The methods for performing such a transformation are similar to those described for the general synthesis of 6.

Additionally, the amine group can be introduced prior to the formation of the sulfonamide and benzisoxazole on the nitrile precursor G9 (where R ¹ , or R ² , or R ³ , or R ⁴ = NO ₂ ), as shown in Scheme 3D of the general synthesis 3.

General Synthesis 9

Scheme 9A illustrates the introduction of a nitrile group into the G25 structure as a substituent that is part of Ar or on the benzisoxazole moiety.

Scheme 9A

The method for such a transformation will be obvious to those skilled in the art and includes, but is not limited to, substitution of SnAr or transition metal catalyzed coupling with a suitable cyanide reagent. The group designated as (X) in the G24 structure may be, but is not limited to, a halogen, triflate, or mesylate and is selected to be suitable for the reaction employed.

Additional preferences

The preferences described below may apply to all aspects of the invention described above, or may relate to one aspect. The preferences may be combined in any combination.

R ¹ , R ² , R ³ and R ⁴

In some embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be H. In some of these embodiments, one of R ¹ , R ² , R ³ and R ⁴ is H. In other of these embodiments, two of R ¹ , R ² , R ³ and R ⁴ are H. In other of these embodiments, three of R ¹ , R ² , R ³ and R ⁴ are H.

In some embodiments, at least one of R ¹ , R ² , R ³ , and R ⁴ is not H.

In some embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be C _1-3 alkyl optionally substituted with:

hydroxy,

C _1-2 alkoxy, optionally substituted with one or more fluoro groups;

_NH2 ,

phenyl,

C _5-6 heteroaryl,

C _1-4 alkylcarbamoyl,

acylamido or

one or more fluoro groups.

In these embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be C _1-3 alkyl. Furthermore, in these embodiments, the C _1-3 alkyl group can be methyl, ethyl or propyl. These groups can be unsubstituted. These groups can be substituted with one or more fluoro groups and can be perfluorinated, such as CF ₃ , C ₂ F ₅ . These groups can be substituted with one, two, three, four or five fluoro groups. In some embodiments, these groups can be substituted with one; one or two; or one, two or three fluoro groups.

If the alkyl group is substituted, the substituent may be selected from:

(i) hydroxy; or

(ii) unsubstituted C _1-2 alkoxy, i.e. methoxy, ethoxy; or C _1-2 alkoxy substituted by one or more fluoro groups, e.g. -OCH ₂ F, -OCH ₂ CF ₃ ; or

(iii) NH ₂ ; or

(iv) phenyl; or

(v) C _5-6 heteroaryl, such as N-pyrazolyl; or

(vi) C _1-4 alkylcarbamoyl, such as NHC(O)Me; or

(vii) acylamido, such as NHCO ₂ Me.

In some embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ may be C _1-3 alkoxy optionally substituted with C _3-6 cycloalkyl or one or more fluoro groups. In these embodiments, the C _1-3 alkoxy group may be methoxy, ethoxy or propyloxy. These groups may be unsubstituted. These groups may be substituted with one or more fluoro groups and may be perfluorinated, such as OCF ₃ , OC ₂ F ₅ . These groups may be substituted with one, two, three, four or five fluoro groups. In some embodiments, these groups may be substituted with one; one or two; or one, two or three fluoro groups. The alkoxy group may be substituted with C _3-6 cycloalkyl, such as cyclopropyl. Thus, the entire group may be OCH ₂ (cyclopropyl).

In some embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be C _3-6 cycloalkyl. In these embodiments, the C _3-6 cycloalkyl group can be cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In particular, the C _3-6 cycloalkyl group can be cyclopropyl.

In some embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ may be a halogen. In these embodiments, the halogen group may be fluorine, chlorine, bromine or iodine.

In some embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be ^CORC , wherein R ^C is selected from NR ^N1 R ^N2 , wherein R ^N1 and R ^N2 are independently selected from H and methyl. In these embodiments, the group can be selected from C(O)NH ₂ , C(O)NHCH ₃ and C(O)N(CH ₃ ) ₂ .

In some embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be cyano, NH ₂ , NO ₂ . In some of these embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be cyano. In other of these embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be NH ₂ . In other of these embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be NO ₂ .

In some embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be phenyl or C _5-6 heteroaryl, wherein these groups are optionally substituted with methyl, cyano, hydroxy or methoxy. In some of these embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be phenyl. In other of these embodiments, at least one of R ¹ , R ² , R ³ and R ⁴ can be C _5-6 heteroaryl, such as oxazolyl, pyrazolyl, triazolyl, pyridyl and pyrimidinyl. The phenyl C _5-6 heteroaryl group can be unsubstituted. In some embodiments, the phenyl group can be substituted with methyl, cyano or methoxy. In some embodiments, the C _5-6 heteroaryl group may be substituted with one or more methyl groups such that the entire group is, for example, dimethylpyrazolyl or N-methylpyrazolyl.

In some embodiments, R ⁴ is methoxy.

In some embodiments, R ⁴ is methoxy, R ² is CH ₂ OCH ₃ or CH ₂ OCH ₂ CH ₃ , and R ¹ and R ³ are H.

In some embodiments, R ⁴ is methoxy, R ² is phenyl optionally substituted with methyl or methoxy, and R ¹ and R ³ are H.

In some embodiments, R ⁴ is methoxy, R ² is C _5-6 heteroaryl optionally substituted with methyl.

In some embodiments, R ⁴ is methoxy and R ¹ , R ² and R ³ are H.

In some embodiments, R ⁴ is chloro, R ² is C _1-3 alkyl or bromo, and R ¹ and R ³ are H.

In some embodiments, R ⁴ is chloro, and R ¹ , R ² , and R ⁴ are H.

In some embodiments, R ³ is C _1-3 alkyl, and R ¹ , R ² and R ⁴ are H.

Ar

Ar is selected from phenyl, naphthyl and C _5-10 heteroaryl groups, which may be unsubstituted or substituted.

In some embodiments, Ar is phenyl.

In some embodiments, Ar is naphthyl.

In some embodiments, Ar is a C _5-10 heteroaryl group. The C _5-10 heteroaryl group can be selected from: quinolinyl, benzothiazolyl, quinoxalinyl, benzooxadiazolyl, benzothiadiazolyl, benzofuran, and benzotriazolyl. In some of these embodiments, Ar is quinolinyl or benzothiazolyl.

In some embodiments, Ar is a group:

.

In some embodiments, the Ar substituent is C _1-4 alkyl optionally substituted with hydroxy, C _1-2 alkoxy, NH ₂ , C _1-4 alkylcarbamoyl, or one or more fluoro groups. In these embodiments, the C _1-4 alkyl group can be methyl, ethyl, propyl, or butyl. These groups can be unsubstituted. These groups can be substituted with one or more fluoro groups and can be perfluorinated, such as CF ₃ , C ₂ F ₅ . If the alkyl group is substituted, the substituent can be selected from:

(i) hydroxy; or

(ii) C _1-2 alkoxy, i.e. methoxy, ethoxy; or

(iii) NH ₂ ; or

(iv) C _1-4 alkylcarbamoyl, such as NHC(O)CH ₃ .

In some embodiments, the Ar substituent is C _3-6 cycloalkyl. In these embodiments, the C _3-6 cycloalkyl group can be cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In particular, the C _3-6 cycloalkyl group can be cyclohexyl.

In some embodiments, the substituent Ar is hydroxy; cyano; NR ^N3 R ^N4 , wherein R ^N3 and R ^N4 are independently selected from H and methyl; or acylamido. In some of these embodiments, the substituent may be hydroxy. In other of these embodiments, the substituent may be cyano. In other of these embodiments, the substituent may be NR ^N3 R ^N4 , wherein R ^N3 and R ^N4 are independently selected from H and methyl, such that the substituent may be NH ₂ , NHCH ₃ , or N(CH ₃ ) ₂ . In other of these embodiments, the substituent may be acylamido, such as NHCO ₂ CH ₃ .

In some embodiments, the Ar substituent is a halogen. In these embodiments, the halogen group can be fluorine, chlorine, bromine, or iodine.

In some embodiments, the Ar substituent is C _1-3 alkoxy optionally substituted with hydroxy, C(O)NH ₂ , C _3-6 cycloalkyl, phenyl, C _5-6 heteroaryl, or one or more fluoro groups. In these embodiments, the C _1-3 alkoxy group can be methoxy, ethoxy, or propyloxy. These groups can be unsubstituted. These groups can be substituted with one or more fluoro groups and can be perfluorinated, such as OCF ₃ , OC ₂ F ₅ . The alkoxy group can be substituted with hydroxyl such that the entire group is, for example, OC ₂ H ₄ OH. The alkoxy group can be substituted with C(O)NH ₂ such that the entire group is, for example, OCH ₂ C(O)NH ₂ . The alkoxy group may be substituted by C _3-6 cycloalkyl, for example cyclopropyl, such that the entire group is, for example, OCH ₂ (cyclopropyl). The alkoxy group may be substituted by phenyl such that the entire group is, for example, benzyloxy. The alkoxy group may be substituted by C _5-6 heteroaryl, for example pyridyl, pyrazolyl such that the entire group is, for example, OCH ₂ (N-methylpyrazolyl) or OCH ₂ (methoxypyridyl).

In some embodiments, the Ar substituent is phenoxy, optionally substituted with fluorine. In some of these embodiments, the substituent may be phenoxy. In other of these embodiments, the substituent may be OC ₆ H ₄ F.

In some embodiments, the Ar substituent is phenyl or C _5-6 heteroaryl. In some of these embodiments, the substituent is phenyl. In other of these embodiments, the substituent may be C _5-6 heteroaryl, such as oxazolyl or N-pyrazolyl.

In some embodiments, the Ar substituent is SF ₅ or SO ₂ CH ₃ . In some of these embodiments, the substituent is SF ₅ . In other of these embodiments, the substituent is SO ₂ Me.

In some embodiments, the substituent Ar is -( _CH2 ) _n -Y-, where Y is O or _CH2 and n is 2 or 3. This substituent is particularly important when Ar is phenyl and forms a partially unsaturated fused ring with the phenyl. Thus, Ar can be tetralinyl (i.e., fused cyclohexane), indanyl (i.e., fused cyclopentane), chromanyl (i.e., fused tetrahydropyran), or dihydrobenzofuranyl.

In some embodiments, the Ar substituent is a C _1-4 alkyl ester. In some of these embodiments, the substituent is C(O)OCH ₃ . In other of these embodiments, the substituent is C(O)OC(CH ₃ ) ₃ .

Some embodiments of Ar can be represented by the formula (Ar-1):

(Ar-1)

where Y is N or CR ^A4 , and Z is N or CR ^A5 ; and

R ^A1 , R ^A2 , R ^A3 , R ^A4 (if present) and R ^A5 (if present) are independently selected from H and optional substituents Ar.

In some embodiments, R ^A2 is ethyl.

In some embodiments, R ^A3 is selected from cycloalkyl; phenoxy; phenyl; C _5-6 heteroaryl; SF ₅ ; and SO ₂ CH ₃ .

In some embodiments, Ar is 5-ethyl-2-methoxyphenyl.

In some embodiments, Ar is 5- _CF3-2 -methoxyphenyl.

In some embodiments, Ar is 2,6-dimethoxyphenyl.

In some embodiments, Ar is quinolinyl. These compounds may exhibit selective activity towards HBO1.

In some embodiments, R ⁴ is methoxy, R ² is selected from CH ₂ O CH ₃ , CH ₂ O CH ₂ CH ₃ and optionally substituted phenyl, and Ar is 2,6-dimethoxybenzene. These compounds may exhibit particular activity toward MOZ and MORF. Compounds in which R ² is selected from CH ₂ OCH ₃ and CH ₂ OCH ₂ CH ₃ may exhibit selective activity toward MOZ and MORF.

In some embodiments, for compounds in which R ¹ , R ² , R ^{3 ,} and R ⁴ are H, Ar is not 4-aminophenyl.

In some embodiments, for compounds wherein R ¹ , R ² , R ³ and R ⁴ are H, Ar is not 2,4,6-trimethylphenyl.

In some embodiments, for compounds wherein R ¹ , R ² , and R ⁴ are H and R ³ is CF ₃ , Ar is not 2-(difluoromethoxy)phenyl.

In some embodiments, for compounds wherein R ¹ , R ² , R ³ , and R ⁴ are H, Ar is not 4-fluoro-3-methylphenyl.

In some embodiments, for compounds wherein R ¹ , R ² , and R ³ are H and R ⁴ is methoxy, Ar is not unsubstituted naphthyl.

Compounds of particular interest include those in the examples.

EXAMPLES

The following examples are provided solely to illustrate the present invention and are not intended to limit the scope of the invention described herein.

Abbreviations

For convenience, many chemical moieties are represented using well-known abbreviations, including, but not limited to, methyl (Me), ethyl (Et), N- propyl (nPr), isopropyl (iPr), N -butyl (nBu), tert -butyl (tBu), phenyl (Ph), benzyl (Bn), methoxy (MeO), ethoxy (EtO), trimethylsilyl (TMS), and acetyl (Ac).

For convenience, many chemical compounds are represented using well-known abbreviations including, but not limited to, methanol (MeOH), deuterated methanol (methanol _- d4 ), ethanol (EtOH), isopropanol ( i -PrOH), ethyl acetate (EtOAc), acetic acid (AcOH), acetonitrile (MeCN or ACN), dichloromethane (methylene chloride, DCM), trifluoroacetic acid (TFA), N,N -dimethylformamide (DMF), tetrahydrofuran (THF), dimethyl sulfoxide (DMSO), N -methyl-2-pyrrolidone (NMP), deuterated acetone (acetone _- d6 ), deuterated chloroform ( _CDCl3 ), deuterated dimethyl sulfoxide (DMSO _- d6 ), 1,1'-bis(diphenylphosphino)ferrocene (dppf), triethylamine (Et ₃ N or TEA), N,N- diisopropylethylamine (DIPEA or DIEA), 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium(II) (PdCl ₂ (dppf)), trans -dichlorobis(triphenylphosphine)palladium(II) (PdCl ₂ (PPh ₃ ) ₂ ), tris(dibenzylideneacetone) dipalladium(0) (Pd ₂ (dba) ₃ ), tetrakis(triphenylphosphine)palladium(0) (Pd(PPh ₃ ) ₄ ), 2,4-dimethoxybenzyl (DMB), petroleum ether (petroleum ether). ether), lithium bis(trimethylsilyl)amide (LHMDS or LiHMDS), potassium bis(trimethylsilyl)amide (KHMDS), sodium bis(trimethylsilyl)amide (NaHMDS), n - butyllithium (n-BuLi), N- bromosuccinimide (NBS), N -chlorosuccinimide (NCS), pyridinium p -toluenesulfonate (PPTS), azobisisobutyronitrile (AIBN), tetramethylethylenediamine (TMEDA), tert- butyldimethylsilyl chloride (TBS-Cl), tetra -n -butylammonium fluoride (TBAF) and diisopropyl azodicarboxylate (DIAD).

In addition, TLC refers to thin layer chromatography.

Other abbreviations: retention time (RT or _RT ), minute(s) (min), hour(s) (hr), room temperature (RT), concentrated (conc), atmosphere (atm), aqueous (aq), saturated (sat), eq (equivalent(s)).

General details of the experiments

Unless otherwise stated, the following conventions apply. ¹ H NMR spectra were recorded on a Bruker Ultrashield Plus (400 MHz) or Bruker AVANCE III (400 MHz). Signal multiplicity is indicated by the following abbreviations: s: singlet; d: doublet; t: triplet; q: quartet; n: pentet; dd: doublet of doublets; dt: doublet of triplets; tt: triplet of triplets; br: broad; m, multiplet. All observed coupling constants, J , are expressed in hertz (Hz). Exchangeable protons are not always observed.

LCMS data were acquired using the following equipment: Agilent 6100 Series Single Quad (LCMS-A), Agilent 1260 Infinity Series LC/MS (LCMS-B), Agilent 1200 (LCMS-C and LCMS-D), Waters 2695 alliance (LCMS-E), Agilent 6120 Single Quad (LCMS-F), or mass-separated LC/MS. Chlorine isotopes are reported as ³⁵ Cl, bromine isotopes are reported as either ⁷⁹ Br or ⁸¹ Br, or both ⁷⁹ Br/ ⁸¹ Br.

LCMS Method A (LCMS-A):

Apparatus: Agilent 6100 Series Single Quad LC/MS

Agilent 1200 Series HPLC

Pump: 1200 Series G1311A Four Channel Pump

Automatic Sampler: 1200 Series G1329A Thermostated Automatic Sampler

Detector: G1314B 1200 Series Variable Wavelength Detector

LC conditions:

Analysis by reversed-phase HPLC

Column: Luna C8 (2) 5 µm 50 × 4.6 mm 100 Å

Column temperature: 30 °C

Injection volume: 5 µl

Solvent A: water, 0.1% formic acid

Solvent B: MeCN, 0.1% formic acid

Gradient: 5-100% solvent B in 10 min

Detection: 254 nm or 214 nm

MS conditions:

Ion source: quadrupole

Ion detection mode: multi-mode with electrospray ionization (ES)

Drying gas temperature: 300 °C

Evaporator temperature: 200 °C

Capillary voltage (V): 2000 (positive)

Capillary voltage (V): 4000 (negative)

Scan range: 100-1000

Step size: 0.1 s

Detection time: 10 min

LCMS Method B (LCMS-B):

Apparatus: Agilent 1260 Infinity series LC/MS

Pump: Dual Channel Pump 1260 Infinity G1312B

Automatic Sampler: 1260 Infinity G1367E 1260 HiP ALS

Detector: Diode Array (DAD) 1290 Infinity G4212A 1290

LC conditions:

Analysis by reversed-phase HPLC

Column: Poroshell 120 EC-C18 2.7 µm 50 × 3.0 mm

Column temperature: 35 °C

Volume of injected sample: 1 µl

Solvent A: water, 0.1% formic acid

Solvent B: MeCN, 0.1% formic acid

Gradient: 5-100% solvent B in 3.8 min

Detection: tracking at 254nm and 214nm

MS conditions:

Ion source: quadrupole

Ion detection mode: atmospheric pressure electrospray ionization (API-ES)

Drying gas temperature: 350 °C

Capillary voltage (V): 3000 (positive)

Capillary voltage (V): 3000 (negative)

Scan range: 100-1000

Step size: 0.1 s

Detection time: 5 min

LCMS, Method C (LCMS-C):

LC chromatograph model: Agilent 1200

(Pump type: dual channel pump, detector type: DAD )

MS Model: Agilent G6110A Quadrupole

LC conditions:

Column: Xbridge-C18, 2.5 µm, 2.1 x 30 mm

Column temperature: 30 °C

Detection wavelength: 214nm, 254nm

Mobile phase: A: 0.07% aqueous HCOOH, B: MeOH

MS conditions:

MS: Ion source: ES+ (or ES-) MS range: 50-900 m/z

Fragmenter: 60 Drying gas flow rate: 10 l/min

Atomizer Pressure: 35 PSI Drying Gas Temperature: 350°C

Vcap: 3.5 kV.

Gradient table:

Flow rate (ml/min) T (min) A (%) B (%) 0.5 0,0 70 30 0.5 0.2 70 30 0.5 1.8 5 95 0.5 2.4 5 95 0.5 2.6 70 30 0.5 3.5 70 30

Sample preparation:

The sample was dissolved in methanol, the concentration was about 0.11-1 mg/mL, then filtered through a 0.22 μm syringe filter. (Injection volume: 1-10 μL)

LCMS, Method D (LCMS-D):

LC chromatograph model: Agilent 1200

(Pump type: dual channel pump, detector type: DAD )

MS Model: Agilent G6110A Quadrupole

LCMS conditions:

LC: Column: Xbridge-C18, 2.5 µm, 2.1 x 30 mm

Column temperature: 30 °C

Detection wavelength: 214nm, 254nm

Mobile phase: A: 0.07% aqueous HCOOH, B: MeOH

MS conditions:

MS: Ion source: ES+ (or ES-) MS range: 50-900 m/z

Fragmenter: 60 Drying gas flow rate: 10 l/min

Atomizer Pressure: 35 PSI Drying Gas Temperature: 350°C

Vcap: 3.5 kV.

Gradient table:

Flow rate (ml/min) T (min) A (%) B (%) 0.5 0,0 70 30 0.5 0.3 70 30 0.5 0.6 50 50 0.5 0.9 40 60 0.5 1,2 30 70 0.5 3.2 5 95 0.5 3.5 5 95 0.5 4.0 70 30 0.5 5.0 70 30

Sample preparation:

The sample was dissolved in methanol, the concentration was about 0.11-1 mg/mL, then filtered through a 0.22 μm syringe filter. (Injection volume: 1-10 μL)

LCMS, Method E (LCMS-E):

Equipment information:

LC model: Waters 2695 alliance

(Pump type: four-channel pump, Detector: photodiode array detector 2996)

MS model: Micromass ZQ

LC conditions:

LC: Column: Xbridge-C18, 3.5 µm, 2.1 x 50 mm

Column temperature: 30 °C

Detection wavelength: 214nm, 254nm

Mobile phase: A: 0.07% aqueous HCOOH, B: MeOH

MS conditions:

MS: Ion source: ES+ (or ES-) MS range: 50-900 m/z

Capillary: 3 kV Cone: 3 V Extractor: 3 V

Drying gas flow rate: 600 l/h Cone: 50 l/h

Desolvation temperature: 300 °C

Source temperature: 100 °C.

Gradient table:

Flow rate (ml/min) T (min) A (%) B (%) 0.3 0,0 80 20 0.3 0.5 80 20 0.3 0.8 50 50 0.3 1,2 35 65 0.3 2.0 20 80 0.3 4.0 5 95 0.3 5.0 5 95 0.3 5.8 15 85 0.3 6.2 80 20 0.3 8.0 80 20

Sample preparation:

The sample was dissolved in methanol, the concentration was about 0.11-1 mg/mL, then filtered through a 0.22 μm syringe filter. (Injection volume: 1-10 μL)

LCMS, Method F (LCMS-F)

Apparatus: Agilent 6120 Series Single Quad LC/MS

Agilent 1200 Series HPLC

Pump: 1200 Series G1311A Four Channel Pump

Automatic Sampler: 1200 Series G1329A Thermostated Automatic Sampler

Detector: G1314B 1200 Series Variable Wavelength Detector

LC conditions:

Analysis by reversed-phase HPLC

Column: Luna C8 (2) 5 µm 50 x 4.6 mm 100 Å

Column temperature: 30 °C

Injection volume: 1-10 µl

Solvent A: water, 0.1% formic acid

Solvent B: MeCN, 0.1% formic acid

Gradient: 0-95% solvent B in 10 min

Detection: 254 nm or 214 nm

MS conditions:

Ion source: quadrupole

Ion detection mode: multi-mode with ES and APCI

Drying gas temperature: 250 °C

Evaporator temperature: 200 °C

Capillary voltage (V): 4000 (positive)

Capillary voltage (V): 4000 (negative)

Scan range: 100-1000

Step size: 0.1 s

Detection time: 10 min

Preparative mass-separated HPLC

Device:

Waters ZQ 3100 - mass detector

Waters 2545 - pump

Waters SFO - pneumatic hydro jet system organizer

Waters 2996 - Diode Array Detector

Waters 2767 Sample Management System

LC conditions:

Analysis by reversed-phase HPLC

Column: XBridge TM C18 5 µm 19 x 50 mm

The volume of the injected sample is 500 µl

Solvent A: water, 0.1% formic acid

Solvent B: acetonitrile, 0.1% formic acid

Gradient: 25-100% B in 10 min

Flow rate: 19 ml/min

Detection: 100-600 nm

MS conditions:

Ion source: single-quadrupole

Ion detection mode: ES-positive.

Source temperature: 150 °C

Desolvation temperature: 350 °C

Detection: Ion Counting

Capillary (KV)-3.00

Cone (B): 30

Extractor (B): 3

RF lens (B): 0.1

Scanning range: 100-1000 a.m.u.

Scan time: 0.5 sec

Detection time: 10 min

Gas flow

Desolvation, l/h - 650

Cone, l/h - 100

Preparative HPLC (prep. HPLC)

Machine type: Varian 940-LC series;

Pump type: four-channel pump;

Detector type: diode array detector.

HPLC conditions:

Waters Sunfire prep C18 OBD, 5 µm 19 x 100 mm column, eluting with a gradient of MeOH in water, with 0.07% TFA at a flow rate of 15 mL/min. Detection wavelength: 214 nm, 254 nm.

Analytical thin-layer chromatography was performed on Merck silica gel 60 F254 aluminum-backed plates visualized by fluorescence quenching under UV light or by immersion in basic KMnO ₄ or ninhydrin.

Preparative thin-layer chromatography (preparative TLC or prep. TLC) was performed using Tklst (China), grand grade: (HPTLC): 8 ± 2 μm >80%; (TLC): 10-40 μm. Type: GF254. Compounds were visualized by UV (254 nm).

Column chromatography was performed using the Biotage Isolera purification system with Grace or RediSep® silica gel cartridges or Tklst (China) grand-grade silica gel with a particle size of 100-200 mesh.

Microwave irradiation was performed using a CEM Explorer SP microwave reactor.

When necessary, anhydrous solvents were purchased from Sigma-Aldrich or dried using conventional methods.

Unless otherwise stated, acidification was performed with concentrated or aqueous HCl solution.

Additional used cartridges:

Phase separator:

Manufacturer: Biotage

Product: ISOLUTE® Phase Separator (unless otherwise stated, 3 ml)

Amine Modified Silica Cartridges:

Manufacturer: Biotage

Product: Isolute® NH2, 1 g/6 ml

or

Manufacturer: Silicycle

Product: Silica gel with amine, 500 mg or 1 g

Synthesis of intermediate compounds

i) 6-(Methoxymethyl)-5-methylbenzo[ d ]isoxazol-3-amine I4

a) methyl 4-cyano-5-fluoro-2-methylbenzoate I1

A mixture of 4-bromo-2-fluoro-5-methylbenzonitrile (3.5 g, 16.4 mmol), Pd(dppf) _Cl2⋅DCM (668 mg, 0.82 mmol), and _Et3N (5.0 g, 49.1 mmol) in MeOH (80 mL) was heated at 100 °C under CO (0.2 MPa) overnight. Additional Pd(dppf) _Cl2⋅DCM (340 mg, 0.4 mmol) was added and heating was continued under CO (0.2 MPa) overnight. The catalyst was removed by filtration, washed with MeOH, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=20/1 to 10/1 to 5/1) to give the title compound (2.4 g, 74%) as a yellow solid. LCMS-D: _sp 2.48 min; m/z 216.1 [M+Na] ⁺ .

b) 2-Fluoro-4-(hydroxymethyl)-5-methylbenzonitrile I2

To a solution of methyl 4-cyano-5-fluoro-2-methylbenzoate I1 (2.4 g, 12.4 mmol) in anhydrous THF (20 mL) at rt under N ₂ was added dropwise LiBH ₄ (2.0 M solution in THF, 12.4 mL, 24.8 mmol) and the mixture was heated at reflux for 2 h. The reaction was quenched with water (80 mL) and the mixture was extracted with EtOAc (90 mL × 3). The combined organic extracts were washed with water (100 mL × 3), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=20/1 to 10/1 to 5/1) to give the title compound (1.6 g, 79%) as a yellow solid. LCMS-D: _sp 1.43 min; m/z 166.1 [M+H] ⁺ , 188.1 [M+Na] ⁺ .

c) 2-Fluoro-4-(methoxymethyl)-5-methylbenzonitrile I3

To a solution of 2-fluoro-4-(hydroxymethyl)-5-methylbenzonitrile I2 (800 mg, 8.8 mmol) and iodomethane (3.4 g, 24.2 mmol) in DMF (12 mL) at 0 °C was added NaH (60 wt % dispersion in oil, 379 mg, 9.7 mmol) and the mixture was stirred at 0 °C for 30 min. Water was added and the mixture was extracted with EtOAc (50 mL × 3). The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=20/1 to 10/1 to 5/1) to give the title compound (660 mg, 76%) as a yellow solid. LCMS-D: _sp 2.44 min; m/z 180.1 [M+H] ⁺ , 202.1.1 [M+Na] ⁺ .

d) 6-(Methoxymethyl)-5-methylbenzo[d]isoxazol-3-amine I4

To a solution of acetohydroxamic acid (792 mg, 10.6 mmol) in anhydrous DMF (20 mL) at 0 °C was added potassium tert -butoxide (1.2 g, 10.6 mmol) and the mixture was stirred at rt for 2 h. Then 2-fluoro-4-(methoxymethyl)-5-methylbenzonitrile I3 (630 mg, 3.5 mmol) was added and the mixture was heated at 60 °C overnight. Water was added and the mixture was extracted with EtOAc (80 mL × 3). The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=20/1 to 10/1) to give the title compound (1.0 g, 77%) as a yellow solid. LCMS-D: _sp 1.75 min; m/z 193.1 [M+H] ⁺ .

ii) 4-Methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I9

a) 4-Bromo-2-fluoro-6-methoxybenzonitrile I5

To a solution of 4-bromo-2,6-difluorobenzonitrile (6.0 g, 27.5 mmol) in THF (100 mL) was added sodium methanolate (1.5 g, 55.0 mmol) and the mixture was stirred at rt for 48 h. Water was added and the mixture was extracted with EtOAc (150 mL x 3). The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=300/1 to 200/1) to give the title compound (4.3 g, 68%) as a white solid. LCMS-D: _sp 2.53 min; m/z 251.8/253.8 [M+Na] ⁺ .

b) Methyl 4-cyano-3-fluoro-5-methoxybenzoate I6

A mixture of 4-bromo-2-fluoro-6-methoxybenzonitrile I5 (4.3 g, 18.7 mmol), Pd(dppf) _Cl2⋅DCM (768 mg, 0.94 mmol) and _Et3N (5.7 g, 56.1 mmol) in MeOH (50 mL) was heated at 100 °C under CO atmosphere (0.2 MPa) overnight. The catalyst was removed by filtration, washed with MeOH and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=200/1 to 50/1) to give the title compound (2.9 g, 74%) as a white solid. LCMS-D: _sp 2.41 min; m/z 210.0 [M+H] ⁺ , 232.0 [M+Na] ⁺ .

c) 2-Fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7

To a solution of LiBH ₄ (2.0 M solution in THF, 13.9 mL, 27.8 mmol) in anhydrous THF (60 mL) at rt under N ₂ was added dropwise a solution of methyl 4-cyano-3-fluoro-5-methoxybenzoate I6 (2.9 g, 13.9 mmol) in anhydrous THF (10 mL) and the mixture was heated at reflux for 1 h. The reaction was quenched with 1 M aq HCl and extracted with EtOAc (100 mL × 3). The combined organic extracts were washed with water (100 mL x 3), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to give the title compound (2.5 g, 100%) as a white solid. LCMS-D: _sp 2.31 min; m/z 182.1 [M+H] ⁺ , 204.1 [M+Na] ⁺ .

d) 2-Fluoro-6-methoxy-4-(methoxymethyl)benzonitrile I8

To a solution of 2-fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7 (2.7 g, 14.9 mmol) and iodomethane (10.6 g, 74.5 mmol) in DMF (100 mL) at 0 °C was added NaH (60 wt % dispersion in oil, 1.2 g, 29.8 mmol) in small portions and the mixture was stirred at rt for 30 min. Water was added and the mixture was extracted with EtOAc (100 mL × 3). The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 5/1) to give the title compound (2.2 g, 76%) as a yellow solid. LCMS-D: _sp 2.22 min; m/z 218.0 [M+Na] ⁺ .

e) 4-Methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I9

To a solution of acetohydroxamic acid (2.3 g, 30.8 mmol) in anhydrous DMF (1500 mL) at rt was added potassium tert -butoxide (3.5 g, 30.8 mmol) and the mixture was stirred at rt for 1 h. Then 2-fluoro-6-methoxy-4-(methoxymethyl)benzonitrile I8 (2.0 g, 10.3 mmol) was added and stirring was continued at rt overnight. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=20/1 to 10/1 to 3/1) to give the title compound (580 mg, 27%) as a yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 6.92 (d, J= 0.8 Hz, 1H), 6.65 (s, 1H), 5.91 (s, 2H), 4.48 (s, 2H), 3.90 (s, 3H), 3.32 (s, 3H, hidden by aqueous peak). LCMS-D: _sp 1.33 min; m/z 209.0 [M+H] ⁺ .

iii) 4-Nitrobenzo[ d ]isoxazol-3-amine I10

To a solution of 2- _fluoro -6-nitrobenzonitrile (1.0 g, 6.17 mmol) in DMF/ _H2O (32 mL/32 mL) were added acetohydroxamic acid (2.78 g, 37.0 mmol) and _K2CO3 (10.23 g, 74.0 mmol), and the mixture was heated at 70 °C for 19 h. Water (200 mL) was added and the mixture was extracted with EtOAc (100 mL × 3). The combined organic extracts were washed with water, brine, dried over _{anhydrous Na2SO4} , filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=30/1 to 1/1) to give the title compound (380 mg, 35%) as a yellow solid. LCMS-D: _sp 2.82 min; m/z 180.1 [M+H] ⁺ .

iv) 4-Methoxy-6-(1-methoxyethyl)benzo[ d ]isoxazol-3-amine I15

a) 4-(1-Ethoxyvinyl)-2-fluoro-6-methoxybenzonitrile I11

To a solution of 4-bromo-2-fluoro-6-methoxybenzonitrile I5 (2.0 g, 8.7 mmol) in THF (40 mL) were added tributyl(1-ethoxyvinyl)stannane (3.4 g, 9.6 mmol), Pd( _PPh3 ) ₄ (201 mg, 0.174 mmol) and LiCl (1.15 g, 27.0 mmol), and the mixture was heated under reflux under _N2 for 48 h. The mixture was diluted with EtOAc and washed successively with water, 5% aqueous ammonium hydroxide solution and brine. The organic layer was dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=200/1) to give the title compound (1.6 g, 84%) as a light yellow solid. LCMS-C: _sp 2.41 min; m/z 222.0 [M+H] ⁺ .

b) 4-Acetyl-2-fluoro-6-methoxybenzonitrile I12

To a solution of 4-(1-ethoxyvinyl)-2-fluoro-6-methoxybenzonitrile I11 (1.0 g, 4.5 mmol) in THF (10 mL) was added 2 M aq. HCl (6.0 mL), and the mixture was stirred at rt for 3 h. The mixture was diluted with diethyl ether and washed with saturated aqueous NaHCO ₃ and water. The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give the title compound (710 mg, 81%) as a white solid. LCMS-C: _sp 1.42 min; m/z 194.0 [M+H] ⁺ .

c) 2-Fluoro-4-(1-hydroxyethyl)-6-methoxybenzonitrile I13

To a solution of 4-acetyl-2-fluoro-6-methoxybenzonitrile I12 (700 mg, 3.6 mmol) in THF (30 mL) was added sodium borohydride (206 mg, 5.4 mmol) and the mixture was stirred at rt overnight. Water was added and the mixture was extracted with EtOAc (30 mL x 3). The combined organic extracts _were dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure to give the title compound (675 mg, 95%) as a colorless oil. LCMS-C: _sp 0.98 min; m/z 196.0 [M+H] ⁺ .

d) 2-Fluoro-6-methoxy-4-(1-methoxyethyl)benzonitrile I14

To a solution of 2-fluoro-4-(1-hydroxyethyl)-6-methoxybenzonitrile I13 (670 mg, 3.4 mmol) and iodomethane (1.5 g, 10.3 mmol) in DMF (20 mL) at 0 °C was added NaH (60 wt % dispersion in oil, 274 mg, 6.8 mmol) in small portions and the mixture was stirred at 0 °C for 2 h. Water was added and the mixture was extracted with EtOAc (40 mL x 3). The combined organic extracts were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (650 mg, 90%) as a light yellow solid. LCMS-C: _sp 1.95 min; m/z 210.0 [M+H] ⁺ .

e) 4-Methoxy-6-(1-methoxyethyl)benzo[d]isoxazol-3-amine I15

To a solution of acetohydroxamic acid (698 mg, 9.3 mmol) in anhydrous DMF (20 mL) at 0 °C was added potassium tert -butoxide (1.04 g, 9.3 mmol) and the mixture was stirred at rt for 1 h. Then a solution of 2-fluoro-6-methoxy-4-(1-methoxyethyl)benzonitrile I14 (650 mg, 3.1 mmol) in anhydrous DMF (10 mL) was added dropwise and the mixture was stirred at rt overnight. Water was added and the mixture was extracted with EtOAc (50 mL × 3). The combined organic extracts were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=300/1 to 200/1) to give the title compound (130 mg, 19%) as a yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 6.90 (s, 1H), 6.64 (s, 1H), 5.92 (s, 2H), 4.39 (q, J= 6.4 Hz, 1H), 3.90 (s, 3H), 3.15 (s, 3H), 1.36 (d, J= 6.4 Hz, 3H). LCMS-C: _sp 0.73 min; m/z 223.0 [M+H] ⁺ .

v) 4-Methoxy-6-phenylbenzo[ d ]isoxazol-3-amine I17

a) 3-Fluoro-5-methoxy-[1,1'-biphenyl]-4-carbonitrile I16

To a solution of 4-bromo-2-fluoro-6-methoxybenzonitrile I5 (6.0 g, 26.1 mmol) and phenylboronic acid (6.36 g, 52.2 mmol) in 1,4-dioxane (200 mL) and water (50 mL) under _N2 were added Pd( _PPh3 ) ₄ (2.99 g, 2.66 mmol) and _Na2CO3 (8.29 g, 78.2 mmol) and _the mixture was heated at 100°C overnight. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 50/1) to give the title compound (5.45 g, 93%) as a yellow solid. LCMS-C: _sp 2.48 min; m/z 228.0 [M+H] ⁺ .

b) 4-Methoxy-6-phenylbenzo[d]isoxazol-3-amine I17

To a solution of acetohydroxamic acid (8.15 g, 23.98 mmol) in anhydrous DMF (200 mL) at 0 °C was added potassium tert -butoxide (5.5 g, 24.0 mmol) and the mixture was stirred at rt for 1 h. Then 3-fluoro-5-methoxy-[1,1'-biphenyl]-4-carbonitrile I16 (5.45 g, 7.99 mmol) was added and the mixture was heated at 60 °C for 4 h. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=10/1 to 8/1 to 6/1) to give the title compound (2.2 g, 38%) as a yellow solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.79-7.74 (m, 2H), 7.52-7.46 (m, 2H), 7.45-7.39 (m, 1H), 7.26 (d, J = 1.1 Hz, 1H), 6.95 (s, 1H), 5.97 (s, 2H), 4.00 (s, 3H). LCMS-C: _sp 2.15 min; m/z 241.0 [M+H] ⁺

vi) 3-(3-Amino-4-methoxybenzo[ d ]isoxazol-6-yl)phenol I19

a) 3-Fluoro-3'-hydroxy-5-methoxy-[1,1'-biphenyl]-4-carbonitrile I18

To a solution of 4-bromo-2-fluoro-6-methoxybenzonitrile I5 (650 mg, 2.8 mmol) and (3-hydroxyphenyl)boronic acid (1.2 g, 5.6 mmol) in 1,4-dioxane (40 mL) and water (10 mL) under _N2 were added Pd( _PPh3 ) ₄ (327 mg, 0.28 mmol) and _Na2CO3 (899 _mg , 8.5 mmol) and the mixture was heated at 100°C overnight. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=3/1) to give the title compound (687 mg, 94%) as a yellow solid. LCMS-C: _sp 2.07 min; m/z 244.0 [M+H] ⁺ .

b) 3-(3-Amino-4-methoxybenzo[d]isoxazol-6-yl)phenol I19

To a solution of acetohydroxamic acid (636 mg, 8.5 mmol) in anhydrous DMF (60 mL) at 0 °C was added potassium tert -butoxide (952 mg, 8.5 mmol) and the mixture was stirred at rt for 1 h. Then 3-fluoro-3'-hydroxy-5-methoxy-[1,1'-biphenyl]-4-carbonitrile I18 (687 mg, 2.8 mmol) was added and the mixture was heated at 60 °C for 4 h. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=1/1) to give the title compound (282 mg, 39%) as a yellow solid. LCMS-C: _sp 2.3 min; m/z 257.0 [M+H] ⁺ .

vii) 6-(Ethoxymethyl)-4-methoxybenzo[ d ]isoxazol-3-amine I21

a) 4-(Ethoxymethyl)-2-fluoro-6-methoxybenzonitrile I20

To a solution of 2-fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7 (1.15 g, 6.4 mmol) and iodoethane (5.0 g, 31.7 mmol) in DMF (40 mL) at 0 °C was added NaH (60 wt % dispersion in oil, 508 mg, 12.7 mmol) in small portions and the mixture was stirred at rt for 30 min. Water was added and the mixture was extracted with EtOAc (100 mL × 3). The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=2/1) to give the title compound (1.0 g, 79%) as a yellow solid. LCMS-C: _sp 2.15 min; m/z 210.0 [M+H] ⁺ .

b) 6-(Ethoxymethyl)-4-methoxybenzo[d]isoxazol-3-amine I21

To a solution of acetohydroxamic acid (1.1 g, 14.3 mmol) in anhydrous DMF (50 mL) at rt was added potassium tert -butoxide (1.6 g, 14.3 mmol) and the mixture was stirred at rt for 1 h. Then 4-(ethoxymethyl)-2-fluoro-6-methoxybenzonitrile I2O (1.0 g, 4.8 mmol) was added and the mixture was stirred at rt overnight. Water was added and the mixture was extracted with EtOAc. The organic extracts were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=20/1 to 10/1 to 3/1) to give the title compound (650 mg, 61%) as a yellow oil. ¹ H NMR (400 MHz, DMSO _-d6 ) δ 6.92 (s, 1H), 6.65 (s, 1H), 5.91 (s, 2H), 4.53 (s, 2H), 3.89 (s, 3H), 3.51 (k, J= 7.0 Hz, 2H), 1.17 (t, J= 7.0 Hz, 3H). JHMS-C: vr. _beat 0.82 min; m/z 223.0 [M+H] ⁺ .

viii) 6-Bromo-4-methoxybenzo[ d ]isoxazol-3-amine I22

To a solution of acetohydroxamic acid (2.0 g, 26.1 mmol) in anhydrous DMF (100 mL) at rt was added potassium tert -butoxide (2.9 g, 26.1 mmol) and the mixture was stirred at rt for 1 h. Then 4-bromo-2-fluoro-6-methoxybenzonitrile I5 (2.0 g, 8.7 mmol) was added and the mixture was stirred at rt overnight. Water was added and the mixture was extracted with EtOAc. The organic extracts were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=2/1) to give the title compound (296 mg, 14%) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.32 (d, J= 1.2 Hz, 1H), 6.90 (d, J= 1.2 Hz, 1H), 6.04 (s, 2H), 3.92 (s, 3H). LCMS-C: _sp 1.4 min; m/z 244.0 [M+H] ⁺ .

ix) 7-Ethoxybenzo[ d ]isoxazol-3-amine I26

a) 2-Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile I23

A mixture of 3-bromo-2-fluorobenzonitrile (3.0 g, 15.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (11.4 g, 45 mmol), potassium acetate (5.9 g, 60.0 mmol), and Pd(dppf) _Cl2 (2.2 g, 3.0 mmol) in DMSO (45 mL) and 1,4-dioxane (15 mL) was heated at 105 °C under _N2 for 3 h. The mixture was diluted with EtOAc (30 mL) and washed with water (30 mL × 3). The organic layer was dried over _{anhydrous Na2SO4} , filtered, concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=20/1) to give the title compound (3.9 g, >100%) as a white solid, which was used directly in the next step.

b) 2-Fluoro-3-hydroxybenzonitrile I24

To a solution of 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile I23 (1.9 g, 7.6 mol) in AcOH (19 mL) under _N2 was added dropwise _H2O2 (30% aqueous solution, _1.9 mL) and the mixture was stirred at rt for 2 h then poured into a mixture of EtOAc and _excess aqueous _Na2SO3 . The layers were separated and the organic layer was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered, concentrated under reduced pressure to give the title compound (650 mg, 62%) as an off-white waxy solid. LCMS-D: _sp 0.93 min; m/z 138.1 [M+H] ⁺ .

c) 3-Ethoxy-2-fluorobenzonitrile I25

To a solution of 2-fluoro-3-hydroxybenzonitrile I24 (360 mg, 2.6 mmol) in DMF (30 _{mL) were added Cs2CO3 (} 4.3 g, 13.1 mmol) and iodoethane (1.0 g, 6.6 mmol) and the mixture was stirred at rt overnight. The mixture was diluted with EtOAc (80 mL) and washed with water (50 mL×3 ₎ . The organic layer was dried over anhydrous _Na2SO4 , filtered, concentrated under reduced pressure to give the title compound (220 mg, 51%) as a yellow solid. LCMS-D: sp _temp 2.31 min; m/z 166.1 [M+H] ⁺ .

d) 7-Ethoxybenzo[d]isoxazol-3-amine I26

To a solution of acetohydroxamic acid (300 mg, 4.0 mmol) in DMF (15 mL) at 0 °C under N ₂ was added potassium tert -butoxide (450 mg, 4.0 mmol) and the mixture was heated at 30 °C for 1 h. A solution of 3-ethoxy-2-fluorobenzonitrile I25 (220 mg, 1.3 mmol) in DMF (10 mL) was added and the mixture was heated at 30 °C overnight. EtOAc (80 mL) was added and the mixture was washed with water (50 mL x 3). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered, concentrated under reduced pressure to give the title compound (170 mg, 70%) as a yellow solid. LCMS-D: _sp 1.68 min; m/z 179.1 [M+H] ⁺ .

x) 7-(Cyclopropylmethoxy)benzo[ d ]isoxazol-3-amine I28

a) 3-(Cyclopropylmethoxy)-2-fluorobenzonitrile I27

To a solution of 2-fluoro-3-hydroxybenzonitrile I24 (360 mg, 2.6 mmol) in DMF (30 _{mL) were added Cs2CO3 (} 4.3 g, 13.1 mmol), KI (87 mg, 0.5 mmol) and (bromomethyl)cyclopropane (880 mg, 6.6 mmol) and the mixture was stirred at rt overnight. EtOAc (80 mL) was added and the mixture was washed with water (50 mL x 3). The organic layer was dried over _{anhydrous Na2SO4} , filtered, concentrated under reduced pressure to give the title compound (150 mg, 30%) as a red solid. LCMS-D: _sp 2.54 min; m/z 192.1 [M+H] ⁺ .

b) 7-(Cyclopropylmethoxy)benzo[d]isoxazol-3-amine I28

Prepared from 3-(cyclopropylmethoxy)-2-fluorobenzonitrile I27 according to the procedure described for 7-ethoxybenzo[ d ]isoxazol-3-amine I26, step d. LCMS-D: sp _temp 2.23 min; m/z 205.1 [M+H] ⁺ .

xi) 6-Ethoxybenzo[ d ]isoxazol-3-amine I32

a) 2-Fluoro-4-((tetrahydro-2H-pyran-2-yl)oxy)benzonitrile I29

To a solution of 2-fluoro-4-hydroxybenzonitrile (20 g, 145.9 mmol) and PPTS (733 mg, 2.9 mmol) in DCM (500 mL) under _N2 was added 3,4-dihydro- 2H -pyran (24.5 g, 292 mmol) and the mixture was stirred at rt overnight. The solvent was removed under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=100/0 to 100/2) to give the title compound (27 g, 83%) as a white solid, which was used directly in the next step.

b) 6-((Tetrahydro-2H-pyran-2-yl)oxy)benzo[d]isoxazol-3-amine I30

To a solution of acetohydroxamic acid (13.7 g, 182.3 mmol) in DMF (60 mL) at 0 °C under N ₂ was added potassium tert -butoxide (20.4 g, 182.3 mmol) and the mixture was stirred at rt for 1 h. Then 2-fluoro-4-((tetrahydro-2 H -pyran-2-yl)oxy)benzonitrile I29 (13.4 g, 60.8 mmol) was added and the mixture was stirred at rt overnight. EtOAc (500 mL) was added and the mixture was washed with water (100 mL × 5). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 5/1) to give the title compound (12.1 g, 85%) as a white solid. LCMS-D: _sp 2.31 min; m/z 235.1 [M+H] ⁺ .

c) 3-Aminobenzo[d]isoxazol-6-ol I31

To a solution of 6-((tetrahydro- 2H -pyran-2-yl)oxy)benzo[ d ]isoxazol-3-amine I30 (3.5 g, 15 mmol) in THF (50 mL) was added 2 M aq. HCl (20 mL) and the mixture was stirred at rt for 3 h. The mixture was diluted with EtOAc (300 mL) and washed with water (2 times). The organic layer was dried over anhydrous _Na2SO4 , filtered, concentrated under reduced pressure to give the title compound (2.1 g, 94%) as _a white solid, which was used directly in the next step.

d) 6-Ethoxybenzo[d]isoxazol-3-amine I32

A mixture _of 3-aminobenzo[ d ]isoxazol-6-ol I31 (300 mg, 2 mmol), _Cs2CO3 (2.0 g, 6 mmol), KI (66 mg, 0.4 mmol) and bromoethane (436 mg, 4 mmol) in DMF (30 mL) was heated at 50 °C under _N2 overnight. The mixture was diluted with EtOAc (300 mL) and washed with water (100 mL×5). The organic layer was dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 5/1) to give the title compound (270 mg, 76%) as a white solid. LCMS-D: _sp 0.37 min; m/z 179.0 [M+H] ⁺ .

xii) 6-(Cyclopropylmethoxy)benzo[ d ]isoxazol-3-amine I33

Prepared from 3-aminobenzo[ d ]isoxazol-6-ol I31 according to the procedure described for 6-ethoxybenzo[ d ]isoxazol-3-amine I32, step d (395 mg, 97%). LCMS-D: sp _temp 2.27 min; m/z 205.1 [M+H] ⁺ .

xiii) 6-(1 H -1,2,3-Triazol-1-yl)benzo[ d ]isoxazol-3-amine I36

a) 4-Azido-2-fluorobenzonitrile I34

A mixture of 4-amino-2-fluorobenzonitrile (2.0 g, 14.7 mmol) in water (4 mL), ACN (32 mL), and concentrated HCl (10 mL) was stirred at rt under N ₂ overnight. NaNO ₂ (2.0 g, 29.4 mmol) was then added portionwise and stirring was continued at rt for 2 h. The mixture was cooled to 0 °C, NaN ₃ (1.9 g, 29.4 mmol) was added portionwise, and stirring was continued at rt for 2 h. Water (50 mL) was added, and most of the organic solvent was removed under reduced pressure. After that, the remaining aqueous mixture was extracted with DCM (50 mL x 4) and the combined organic extracts were washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/0 to 20/1) to give the title compound (1.5 g, 62%) as a yellow solid, which was used directly in the next step.

b) 2-Fluoro-4-(1H-1,2,3-triazol-1-yl)benzonitrile I35

A mixture of 4-azido-2-fluorobenzonitrile I34 (500 mg, 3.1 mmol), ethynyltrimethylsilane (454 mg, 4.6 mmol) and CuI (704 mg, 3.7 mmol) in THF (50 mL) was heated at 50 °C under N ₂ for 24 h. Additional ethynyltrimethylsilane (454 mg, 4.6 mmol) was added and the mixture was heated at 50 °C for an additional 24 h, then concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 10/1) to give 2-fluoro-4-(5-(trimethylsilyl)-1H - 1,2,3-triazol-1-yl)benzonitrile (410 mg), which was dissolved in 1 M TBAF in THF (50 mL) and heated at 45 °C under N ₂ atmosphere overnight. The solvent was removed under reduced pressure, and the residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 2/1) to give the title compound (200 mg, 34%) as a white solid, which was used directly in the next step.

c) 6-(1H-1,2,3-Triazol-1-yl)benzo[d]isoxazol-3-amine I36

To a solution of acetohydroxamic acid (239 mg, 3.16 mmol) in DMF (25 mL) at 0 °C under N ₂ was added potassium tert -butoxide (357 mg, 3.18 mmol) and the mixture was stirred at rt for 2 h. Then a solution of 2-fluoro-4-(1 H -1,2,3-triazol-1-yl)benzonitrile I35 (200 mg, 1.06 mmol) in DMF (15 mL) was added and stirring was continued at rt overnight. EtOAc (100 mL) was added and the mixture was washed with water (5 times). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 2/1) to give the title compound (150 mg, 70%) as a white solid. LCMS-D: _sp 0.47 min; m/z 202.1 [M+H] ⁺ .

xiv) 6-(Pyrimidin-2-yl)benzo[ d ]isoxazol-3-amine I39

a) 2-Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile I37

A mixture of 4-bromo-2-fluorobenzonitrile (1.0 g, 5.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (1.3 g, 5.0 mmol), potassium acetate (5.9 g, 20.0 mmol) and Pd(dppf) _Cl2 (2.0 g, 1.0 mmol) in DMSO (50 mL) and 1,4-dioxane (10 mL) was heated at 105 °C under _N2 for 3 h. The mixture was diluted with EtOAc (200 mL) and washed with water (100 mL × 3). The organic layer was dried over _{anhydrous Na2SO4} , filtered, concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=100/0 to 50/1) to give the title compound (1.1 g, 89%) as a white solid, which was used directly in the next step.

b) 2-Fluoro-4-(pyrimidin-2-yl)benzonitrile I38

To a solution of 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile I37 (464 mg, 2 mmol) and 2-bromopyrimidine (736 mg, 4 mmol) in water (40 mL), toluene (40 mL) and i -PrOH (10 mL) under N ₂ atmosphere were added Pd(dppf)Cl ₂ (146 mg, 0.2 mmol) and K _{3PO 4} 3H ₂ O (1.33 g, 5.0 mmol) and the mixture was heated at 85 °C for 4 h. The mixture was diluted with EtOAc (200 mL) and washed with water (50 mL). The organic layer was dried over _{anhydrous Na2SO4} , filtered, concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 2/1) to give the title compound (270 mg, 68%) as a white solid. LCMS-D: _sp 2.38 min; m/z 200.1 [M+H] ⁺ .

c) 6-(Pyrimidin-2-yl)benzo[d]isoxazol-3-amine I39

To a solution of acetohydroxamic acid (306 mg, 4.07 mmol) in DMF (20 mL) at 0 °C under N ₂ was added potassium tert -butoxide (457 mg, 4.07 mmol) and the mixture was heated at 30 °C for 1 h. Then a solution of 2-fluoro-4-(pyrimidin-2-yl)benzonitrile I38 (270 mg, 1.36 mmol) in DMF (10 mL) was added and heating was continued at 30 °C overnight. The mixture was diluted with EtOAc (100 mL) and washed with water (50 mL × 3). The organic layer was dried over _{anhydrous Na2SO4} , filtered, concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 2/1) to give the title compound (200 mg, 69%) as a white solid. LCMS-D: _sp 0.38 min; m/z 213.1 [M+H] ⁺ , 235.1 [M+Na] ⁺ .

xv) 5-Bromobenzo[ d ]isoxazol-3-amine I40

To a solution of acetohydroxamic acid (23.7 g, 0.315 mol) in DMF (800 mL) at 0 °C under N ₂ was added t -BuOK (35.4 g, 0.315 mol) and the mixture was stirred at 15 °C for 2 h. Then 5-bromo-2-fluorobenzonitrile (21.0 g, 0.105 mol) was added and the mixture was stirred at rt overnight. The mixture was diluted with EtOAc (1.5 L) and washed with water (400 mL × 4). The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 3/1) to give the title compound (19 g, 86%) as a white solid. LCMS-D: _sp 2.13 min; m/z 212.9/214.9 [M+H] ⁺ .

xvi) 4-Bromobenzo[ d ]isoxazol-3-amine I41

To a solution of acetohydroxamic acid (11.25 g, 0.15 mol) in DMF (220 mL) at 0 °C under N ₂ was added t -BuOK (16.8 g, 0.15 mol) and the mixture was stirred at 25 °C for 1 h. Then a solution of 2-bromo-6-fluorobenzonitrile (10.0 g, 0.05 mol) in DMF (80 mL) was added dropwise and stirring was continued at 25 °C overnight. The mixture was diluted with water (200 mL) and extracted with EtOAc (400 mL). The organic extract was washed with water (400 mL × 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (7.0 g, 66%) as a light red solid. LCMS-D: _sp 2.05 min; m/z 212.9/214.9 [M+H] ⁺ .

xvii) 4-(Trifluoromethyl)benzo[ d ]isoxazol-3-amine I42

To a solution of acetohydroxamic acid (2.25 g, 30 mmol) in DMF (80 mL) at 0 °C under N ₂ was added t -BuOK (3.37 g, 30 mmol) and the mixture was heated at 30 °C for 1 h. Then a solution of 2-fluoro-6-(trifluoromethyl)benzonitrile (1.89 g, 10 mmol) in DMF (20 mL) was added and heating was continued at 30 °C overnight. The mixture was partitioned between EtOAc (300 mL) and water (100 mL), the layers were separated and the organic layer was washed with water (100 mL × 3), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 1/1) to give the title compound (1.3 g, 64%) as a white solid. LCMS-D: _sp 2.19 min; m/z 203.0 [M+H] ⁺ .

xviii) 5-Bromo-4-chlorobenzo[ d ]isoxazol-3-amine I44

a) 3-Bromo-2-chloro-6-fluorobenzonitrile I43

To a solution of 2-chloro-6-fluorobenzonitrile (1.0 g, 6.4 mmol) in trifluoromethanesulfonic acid (10 mL) at 0 °C under N ₂ was added NBS (1.1 g, 6.4 mmol) and the mixture was stirred at rt overnight. The mixture was poured onto ice and extracted with EtOAc (30 mL × 2). The combined organic extracts were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc = 200/1 to 100/1) to give the title compound (705 mg, 47%) as a white solid, which was used directly in the next step.

b) 5-Bromo-4-chlorobenzo[d]isoxazol-3-amine I44

To a solution of acetohydroxamic acid (5.1 g, 67.8 mmol) in DMF (150 mL) at 0 °C under N ₂ was added t -BuOK (7.6 g, 6.4 mmol) and the mixture was stirred at rt for 2 h. Then 3-bromo-2-chloro-6-fluorobenzonitrile I43 (5.3 g, 22.6 mmol) was added and the mixture was stirred at rt overnight. The mixture was diluted with EtOAc (500 mL) and washed with water (3 times), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 2/1) to give the title compound (3.1 g, 52%) as a white solid, which was used directly in the next step.

xix) 5-Bromo-4-methoxybenzo[ d ]isoxazol-3-amine I48

a) 3-Bromo-6-fluoro-2-methoxybenzoic acid I45

To a solution of diisopropylamine (5.4 g, 53.7 mmol) in THF (150 mL) at -78 °C under N ₂ was added n- BuLi (2.5 M solution in hexanes, 23.4 mL, 58.5 mmol) dropwise and the mixture was stirred at -78 °C for 1 h. The resulting mixture was added dropwise to a solution of 1-bromo-4-fluoro-2-methoxybenzene (10.0 g, 48.8 mmol) in THF (50 mL) at -78 °C and stirring was continued for 90 min. CO ₂ was bubbled through the mixture with stirring at -78 °C for 20 min, then it was allowed to warm to rt and stirred for 15 min. The reaction mixture was adjusted to pH=1 with HCl, diluted with water and extracted with DCM (500 mL). The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=100/1 to 30/1) to give the title compound (8.0 g, 66%) as a colorless oil. LCMS-D: sp _temp 2.12 min; m/z 248.9/250.9 [M+H] ⁺ .

b) 3-Bromo-6-fluoro-2-methoxybenzamide I46

A mixture of 3-bromo-6-fluoro-2-methoxybenzoic acid I45 (8.0 g, 32.1 mmol) and _SOCl2 (30 mL) was heated at 85 °C for 3 h. The mixture was concentrated under reduced pressure, the residue was dissolved in DCM (5 mL) and added dropwise to conc. _NH4OH (20 mL) at 0 °C. The mixture was allowed to warm to rt, stirred for 20 min and then extracted with DCM (50 mL × 3). The combined organic extracts were washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 1/1) to give the title compound (6.8 g, 80%) as a white solid. LCMS-E: _sp 2.24 min; m/z 247.8/249.8 [M+H] ⁺

c) 3-Bromo-6-fluoro-2-methoxybenzonitrile I47

A mixture of 3-bromo-6-fluoro-2-methoxybenzamide I46 (6.8 g, 25.6 mmol) and _SOCl2 (30 mL) was heated at 80 °C overnight and then concentrated under reduced pressure. The residue was partitioned between water and EtOAc, the phases were separated and the organic layer was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 20/1) to give the title compound (3.5 g, 55%) as a colorless oil, which was used directly in the next step.

d) 5-Bromo-4-methoxybenzo[d]isoxazol-3-amine I48

To a solution of acetohydroxamic acid (3.4 g, 45.7 mmol) in DMF (150 mL) at 0 °C under N ₂ was added t -BuOK (5.1 g, 45.7 mmol) and the mixture was stirred at rt for 90 min. Then a solution of 3-bromo-6-fluoro-2-methoxybenzonitrile I47 (3.5 g, 15.2 mmol) in DMF (30 mL) was added and the mixture was heated at 70 °C overnight. The mixture was diluted with EtOAc (1000 mL) and washed with water (3 times), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 3/1) to give the title compound (3.2 g, 86%) as a white solid. LCMS-D: _sp 2.24 min; m/z 243.0/244.9 [M+H] ⁺ .

xx) 4-(Methoxymethyl)benzo[ d ]isoxazol-3-amine I51

a) 2-(Bromomethyl)-6-chlorobenzonitrile I49

A mixture of 2-chloro-6-methylbenzonitrile (2.0 g, 13.2 mmol), NBS (2.5 g, 13.8 mmol) and AIBN (660 mg, 4.0 mmol) in _CCl4 (60 mL) was heated at 85 °C under _N2 overnight. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 50/1) to give the title compound (1.7 g, 37%) as a white solid, which was used directly in the next step.

b) 2-Chloro-6-(methoxymethyl)benzonitrile I50

Sodium metal (115 mg, 4.8 mmol) was dissolved in MeOH (5 mL) and THF (5 mL) and the mixture was stirred at rt for 20 min. Then 2-(bromomethyl)-6-chlorobenzonitrile I49 (560 mg, 2.4 mmol) was added and the mixture was stirred at rt for 5 h. The solvent was removed under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=50/1) to give the title compound (340 mg, 77%) as a colorless oil. ¹ H NMR (400 MHz, DMSO -d ₆ ) δ 7.75-7.70 (m, 3H), 4.82 (s, 2H), 3.35 (s, 3H).

c) 4-(Methoxymethyl)benzo[d]isoxazol-3-amine I51

To a solution of acetohydroxamic acid (422 mg, 5.6 mmol) in DMF (25 mL) at -78 °C under N ₂ was added t -BuOK (630 mg, 5.6 mmol) and the mixture was stirred at 0 °C for 1 h. Then 2-chloro-6-(methoxymethyl)benzonitrile I50 (340 mg, 1.9 mmol) was added and the mixture was stirred at rt overnight and then heated at 85 °C overnight. The mixture was diluted with water (70 mL) and extracted with EtOAc (100 mL × 2). The combined organic extracts were washed with water (200 mL × 3), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOA=5/1 to 3/1) to give the title compound (105 mg, 31%) as a light yellow oil. LCMS-D: _sp 1.57 min; m/z 179.1 [M+H] ⁺ .

xxi) 4-Ethoxybenzo[ d ]isoxazol-3-amine I53

a) 2-Ethoxy-6-fluorobenzonitrile I52

A mixture of 2-fluoro-6-hydroxybenzonitrile (2.0 g, 14.6 mmol), _K2CO3 (6.04 g, 43.8 mmol) and bromoethane (2.38 g, 21.9 mmol) in DMF (4 mL) was stirred at rt under _N2 overnight. _The mixture was diluted with EtOAc (300 mL), washed with water (100 mL×5), brine, dried over _Na2SO4 , _filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 5/1) to give the title compound (1.6 g, 67%) as a white solid. LCMS-E: _sp 5.24 min; m/z 166.1 [M+H] ⁺ .

b) 4-Ethoxybenzo[d]isoxazol-3-amine I53

To a solution of acetohydroxamic acid (2.18 g, 29 mmol) in DMF (40 mL) at 0 °C under N ₂ was added t -BuOK (3.26 g, 29 mmol) and the mixture was stirred at rt for 1 h. Then 2-ethoxy-6-fluorobenzonitrile I52 (1.6 g, 9.7 mmol) was added and the mixture was stirred at rt overnight. The mixture was diluted with DCM (80 mL), washed with water (60 mL x 4), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc = 5/1) to give the title compound (240 mg, 15%) as a white solid. LCMS-E: temp. _spec. 5.05 min; m/z 179.0 [M+H] ⁺ .

xxii) 5-Methoxybenzo[ d ]isoxazol-3-amine I54

To a solution of acetohydroxamic acid (1.49 mg, 19.8 mmol) in DMF (35 mL) at 0 °C under N ₂ was added t -BuOK (2.23 mg, 19.8 mmol) and the mixture was heated at 30 °C for 1 h. Then a solution of 2-fluoro-5-methoxybenzonitrile (1.0 g, 6.6 mmol) in DMF (5 mL) was added and the mixture was heated at 30 °C overnight. The mixture was diluted with water (70 mL) and extracted with EtOAc (100 mL × 3). The combined organic extracts were washed with water (200 mL x 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (110 mg, 11%) as a yellow solid, which was used directly in the next step.

xxiii) 5-Ethoxybenzo[ d ]isoxazol-3-amine I57

a) 2-Fluoro-5-hydroxybenzonitrile I55

A mixture of 2-fluoro-5-methoxybenzonitrile (1.7 g, 1.2 mmol) and pyridine HCl (17 g) was heated at 80 °C under N ₂ for 5 h and then diluted with DCM (40 mL) and washed with 2 M aq. HCl (8 mL) and water (2 × 40 mL). The organic layer was extracted with aqueous K ₂ CO ₃ (50 mL × 2), the combined aqueous extracts were washed with DCM (70 mL × 2), then the pH was adjusted to 3-4 with 2 M aq. HCl and extracted with DCM (80 mL × 3). The combined organic extracts _were dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure to give the title compound (430 mg, 28%) as an off-white solid, which was used directly in the next step.

b) 5-Ethoxy-2-fluorobenzonitrile I56

To a solution of 2-fluoro-5-hydroxybenzonitrile I55 (430 mg, 3.1 mmol) in DMF (15 _mL ) was added _K2CO3 (1.3 g, 9.4 mmol) and the mixture was stirred at rt under _N2 for 30 min. Then bromoethane (512 mg, 4.7 mmol) was added and stirring was continued at rt overnight. The mixture was diluted with water (70 mL) and extracted with EtOAc (100 mL × 2). The combined organic extracts were washed with water (200 mL × ₃ ), dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=30/1 to 20/1) to give the title compound (480 mg, 92%) as a white solid. LCMS-D: _sp 2.44 min; m/z 166.0 [M+H] ⁺ 188.0 [M+Na] ⁺ .

c) 5-Ethoxybenzo[d]isoxazol-3-amine I57

To a solution of acetohydroxamic acid (645 mg, 8.7 mmol) in DMF (35 mL) at 0 °C under N ₂ was added t -BuOK (978 mg, 8.7 mmol) and the mixture was heated at 30 °C for 1 h. Then a solution of 5-ethoxy-2-fluorobenzonitrile I56 (480 mg, 2.9 mmol) in DMF (5 mL) was added and the mixture was heated at 30 °C overnight. The mixture was diluted with water (60 mL) and extracted with EtOAc (80 mL × 2). The combined organic extracts were washed with water (150 mL x 2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (400 mg, 77%) as a light yellow solid. LCMS-D: _sp 2.02 min; m/z 179.1 [M+H] ⁺ .

xxiv) 6-(3,5-Dimethyl-1 H -pyrazol-1-yl)benzo[ d ]isoxazol-3-amine I59

a) 2-Chloro-4-(3,5-dimethyl-1H-pyrazol-1-yl)benzonitrile I58

A mixture of 3,5-dimethyl- 1H- pyrazole (5 g, 0.052 mol), NaH (60% dispersion in oil, 2.6 g, 0.065 mol) in DMF (50 mL) was stirred at rt for 1 h. Then a solution of 2-chloro-4-fluorobenzonitrile (6.74 g, 0.043 mol) in DMF (50 mL) was added and stirring was continued at rt for 1 h. The reaction was quenched with water and the mixture was extracted with EtOAc. The organic extract was concentrated under reduced pressure to give the title compound (11.0 g, 92%) as a yellow solid. LCMS-D: _sp 2.58 min; m/z 232.1 [M+H] ⁺ .

b) 6-(3,5-Dimethyl-1H-pyrazol-1-yl)benzo[d]isoxazol-3-amine I59

To a solution of acetohydroxamic acid (972 mg, 12.9 mmol) in DMF (20 mL) at 0 °C under N ₂ was added t -BuOK (1.45 g, 12.9 mmol) and the mixture was heated at 30 °C for 1 h. Then 2-chloro-4-(3,5-dimethyl-1 H -pyrazol-1-yl)benzonitrile I58 (1 g, 4.3 mmol) was added and the mixture was heated at 60 °C for 5 h. The mixture was diluted with water and extracted with EtOAc. The organic extract was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (MeOH/DCM=1/20) to give the title compound (150 mg, 15%) as a white solid. LCMS-D: _sp 2.22 min; m/z 229.1 [M+H] ⁺ .

xxv) 5-Methylbenzo[ d ]isoxazol-3-amine I60

To a solution of acetohydroxamic acid (8.33 g, 0.11 mol) in DMF (200 mL) was added t -BuOK (12.5 g, 0.11 mol) and the mixture was stirred at rt for 1 h. Then 2-fluoro-5-methylbenzonitrile (5 g, 0.37 mol) was added and the mixture was heated at 60 °C overnight. The mixture was diluted with water and extracted with EtOAc. The organic extract was concentrated under reduced pressure and the residue was purified by silica gel chromatography (DCM/MeOH = 200/1 to 50/1) to give the title compound (3.0 g, 55%) as a white solid. LCMS-D: temp. _spec. 1.75 min, m/z 149.0 [M+H] ⁺ .

xxvi) 6-(Methoxymethyl)benzo[ d ]isoxazol-3-amine I62

a) 2-Fluoro-4-(methoxymethyl)benzonitrile I61

A mixture of MeI (2.0 g, 13.2 mmol) and NaH (60% suspension in oil, 790 mg, 19.8 mmol) in THF (50 mL) was stirred at 0 °C for 10 min and then 2-fluoro-4-(hydroxymethyl)benzonitrile (2.0 g, 13.2 mmol) was added and the mixture was stirred at rt for 2 h. The reaction was quenched with water and the mixture was extracted with EtOAc. The organic extract was concentrated under reduced pressure and the residue was purified by silica gel chromatography (pet. ether/EtOAc=100/1 to 20/1) to give the title compound (1.7 g, 78%) as a white solid. LCMS-D: _sp 2.01 min; m/z 166.0 [M+H] ⁺ 187.9 [M+Na] ⁺ .

b) 6-(Methoxymethyl)benzo[d]isoxazol-3-amine I62

To a solution of acetohydroxamic acid (1.5 g, 9.1 mmol) in DMF (50 mL) was added t -BuOK (3.06 g, 27.2 mmol) and the mixture was stirred at rt for 1 h. Then 2-fluoro-4-(methoxymethyl)benzonitrile I61 (1.5 g, 9.1 mmol) was added and the mixture was heated at 40 °C overnight. The mixture was diluted with water and extracted with EtOAc. The organic extract was concentrated under reduced pressure and the residue was purified by silica gel chromatography (pet. ether/EtOAc=100/1 to 10/1) to give the title compound (1 g, 62%) as a yellow solid. LCMS-D: temp. _spec. 0.95 min, m/z 179.0 [M+H] ⁺ .

xxvii) 5-(Trifluoromethoxy)benzo[ d ]isoxazol-3-amine I63

To a solution of acetohydroxamic acid (2.2 g, 0.029 mol) in DMF (50 mL) was added t -BuOK (3.28 g, 0.029 mol) and the mixture was stirred at rt for 1 h. Then 2-fluoro-5-(trifluoromethoxy)benzonitrile (2 g, 9.75 mmol) was added and the mixture was heated at 60 °C overnight. The mixture was diluted with water and extracted with EtOAc. The organic extract was concentrated under reduced pressure to give the title compound (1.6 g, 75%) as a yellow solid. LCMS-D: sp _temp 2.43 min; m/z 219.0 [M+H] ⁺ .

xxviii) 5-Methyl-6-(oxazol-2-yl)benzo[ d ]isoxazol-3-amine I65

a) 2-Fluoro-5-methyl-4-(oxazol-2-yl)benzonitrile I64

To a solution of oxazole (90 mg, 1.31 mmol) in THF (10 mL) at -70 °C under N ₂ was added n- BuLi (2.5 M solution in hexane, 1.1 mL, 2.66 mmol) and the mixture was stirred for 10 min. Solid ZnCl ₂ (380 mg, 2.79 mmol) was added and the mixture was allowed to warm to rt. 4-Bromo-2-fluoro-5-methylbenzonitrile (200 mg, 0.93 mmol) was added and the mixture was heated at 60 °C overnight. The solvent was removed under reduced pressure and the residue was purified by silica gel chromatography (pet. ether/EtOAc 10/1) to give the title compound (50 mg, 27%) as a white solid. LCMS-D: _sp 2.51 min; m/z 203.0 [M+H] ⁺ .

b) 5-Methyl-6-(oxazol-2-yl)benzo[d]isoxazol-3-amine I65

To a solution of acetohydroxamic acid (189 mg, 2.52 mmol) in DMF (10 mL) at 0 °C under N ₂ atmosphere was added t -BuOK (377 mg, 3.26 mmol), and the mixture was stirred at 0 °C for 1 h. Then, 2-fluoro-5-methyl-4-(oxazol-2-yl)benzonitrile I64 (170 mg, 0.84 mmol) was added, and the mixture was heated at 50 °C overnight. The mixture was diluted with water and extracted with EtOAc. The organic extract was dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (DCM/MeOH = 10/1) to give the title compound (90 mg, 50%) as a white solid. LCMS-D: _sp 2.10 min; m/z 216.0 [M+H] ⁺ .

xxix) 7-Bromobenzo[ d ]isoxazol-3-amine I66

To a solution of acetohydroxamic acid (3.75 g, 0.05 mol) in DMF (60 mL) at 0 °C was added t -BuOK (5.6 g, 0.05 mol) and the mixture was stirred at rt for 1 h. Then, a solution of 3-bromo-2-fluorobenzonitrile (5.0 g, 0.025 mol) in DMF (90 mL) was added dropwise and stirring was continued at rt overnight. The mixture was diluted with DCM (300 mL), washed with water (250 mL x 4), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (4.0 g, 63%) as a white solid. LCMS-D: temp. _spec. 2.09 min, m/z 213.0/215.0 [M+H] ⁺ .

xxx) 7-(Pyrimidin-2-yl)benzo[ d ]isoxazol-3-amine I68

a) 2-Fluoro-3-(pyrimidin-2-yl)benzonitrile I67

A mixture of 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile I23 (1.5 g, 6.1 mmol), 2-bromopyrimidine (1.9 g, 12.0 mmol), Pd(dppf) _Cl2 (1.3 g, 1.8 mmol) and _{K3PO4 (} 6.5 g, 24.2 mmol) in water (60 mL), toluene (60 mL) and i -PrOH (15 mL) was heated at 85 °C under _N2 for 4 h. The mixture was diluted with EtOAc (50 mL) and washed with water (80 mL × 3). The organic layer was dried over _{anhydrous Na2SO4} , filtered, concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=4/1) to give the title compound (450 mg, 38%) as a light yellow solid. LCMS-E: _sp 4.82 min; m/z 199.9 [M+H] ⁺ .

b) 7-(Pyrimidin-2-yl)benzo[d]isoxazol-3-amine I68

To a solution of acetohydroxamic acid (243 mg, 3.2 mmol) in DMF (15 mL) at 0 °C under N ₂ was added t -BuOK (363 mg, 3.2 mmol), and the mixture was stirred for 1 h. Then, a solution of 2-fluoro-3-(pyrimidin-2-yl)benzonitrile I67 (400 mg, 1.6 mmol) in DMF (5 mL) was added dropwise, and the mixture was stirred at rt overnight. The mixture was diluted with EtOAc (80 mL) and washed with water (60 mL x 3). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give the title compound (230 mg, 67%) as a yellow solid. LCMS-D: temp. _spec. 0.80 min, m/z 213.1 [M+H] ⁺ .

xxxi) 6-(1 H -Pyrazol-1-yl)benzo[ d ]isoxazol-3-amine I70

a) 2-fluoro-4-(1H-pyrazol-1-yl)benzonitrile I69

A mixture of 4-bromo-2-fluorobenzonitrile (400 mg, 2.0 mmol), 1 H -pyrazole (177 mg, 2.6 mmol), CuI (381 mg, 2.0 mmol), K ₃ PO ₄ (849 mg, 4.0 mmol), and (1 S ,2 S )- N ¹ , N ² -dimethylcyclohexane-1,2-diamine (28 mg, 0.2 mmol) in DMF (20 mL) was heated at 100 °C in a microwave reactor for 1 h. The mixture was partitioned between EtOAc (200 mL) and water (100 mL), the layers were separated, and the organic layer was washed with water (3 times), brine, dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 20/1) to give the title compound (120 mg, 32%) as a white solid. LCMS-D: _sp 2.20 min, m/z 188.1 [M+H] ⁺ .

b) 6-(1H-Pyrazol-1-yl)benzo[d]isoxazol-3-amine I70

To a solution of acetohydroxamic acid (215 mg, 2.9 mmol) in DMF (25 mL) at 0 °C was added t -BuOK (322 mg, 2.9 mmol) and the mixture was heated at 30 °C for 2 h. Then 2-fluoro-4-(1 H -pyrazol-1-yl)benzonitrile I69 (120 mg, 0.64 mmol) was added and the mixture was heated at 30 °C overnight. The mixture was partitioned between EtOAc (100 mL) and water (50 mL), the layers were separated and the organic layer was washed with water (3 times), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 2/1) to give the title compound (72 mg, 57%) as a white solid, which was used directly in the next step.

xxxii) 6-(2 H -1,2,3-Triazol-2-yl)benzo[ d ]isoxazol-3-amine I72

a) 2-Chloro-4-(2H-1,2,3-triazol-2-yl)benzonitrile I71

A mixture of 2 H -1,2,3-triazole (553 mg, 8.0 mmol) and NaH (60% dispersion in oil, 192 mg, 4.8 mmol) in DMF (20 mL) was stirred at 0 °C for 30 min, after which a solution of 2-chloro-4-fluorobenzonitrile (622 mg, 4.0 mmol) in DMF (10 mL) was added. The mixture was stirred at 0 °C for 2 h, then allowed to warm to rt and stirred for 2 h. The mixture was partitioned between EtOAc (300 mL) and water (100 mL), the layers were separated and the organic layer was washed with water (3 times), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 20/1) to give the title compound (200 mg, 24%) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.31-8.28 (m, 3H), 8.19-8.14 (m, 2H).

b) 6-(2H-1,2,3-Triazol-2-yl)benzo[d]isoxazol-3-amine I72

To a solution of acetohydroxamic acid (221 mg, 2.9 mmol) in DMF (25 mL) at 0 °C was added t -BuOK (330 mg, 2.9 mmol) and the mixture was heated at 30 °C for 2 h. Then 2-chloro-4-(2 H -1,2,3-triazol-2-yl)benzonitrile I71 (200 mg, 0.98 mmol) was added and the mixture was heated at 30 °C overnight. The mixture was partitioned between EtOAc (100 mL) and water (50 mL), the layers were separated and the organic layer was washed with water (3 times), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 2/1) to give the title compound (90 mg, 46%) as a white solid. LCMS-D: _sp 1.93 min, m/z 202.1 [M+H] ⁺ .

xxxiii) 6-(Pyridin-2-yl)benzo[ d ]isoxazol-3-amine I74

a) 2-Fluoro-4-(pyridin-2-yl)benzonitrile I73

A mixture of 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile I37 (494 mg, 2.0 mmol), 2-bromopyridine (948 mg, 6.0 mmol), Pd(dppf) _Cl2 (293 mg, 0.4 mmol) and _{K3PO4 3H2O} (2.66 g, 10.0 mmol) in _H2O (40 mL), toluene (40 mL) and i -PrOH (10 mL) was _heated at 85°C under _N2 for 4 h. The mixture was partitioned between EtOAc (200 mL) and water (30 mL), the layers were separated and the organic layer was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=30/1 to 10/1) to give the title compound (190 mg, 48%) as a white solid. LCMS-D: _sp 2.32 min, m/z 199.1 [M+H] ⁺ .

b) 6-(Pyridin-2-yl)benzo[d]isoxazol-3-amine I74

To a solution of acetohydroxamic acid (216 mg, 2.88 mmol) in DMF (50 mL) at 0 °C was added t -BuOK (323 mg, 2.88 mmol) and the mixture was stirred at rt for 1 h. Then a solution of 2-fluoro-4-(pyridin-2-yl)benzonitrile I73 (190 mg, 0.96 mmol) in DMF (10 mL) was added and the mixture was stirred at rt overnight. The mixture was partitioned between EtOAc (200 mL) and water (50 mL), the layers were separated and the organic layer was washed with water (50 mL × 3), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=100/0 to 100/1) to give the title compound (105 mg, 52%) as a white solid. LCMS-D: _sp 0.83 min, m/z 212.1 [M+H] ⁺ .

xxxiv) 6-Bromobenzo[ d ]isoxazol-3-amine I75

To a solution of acetohydroxamic acid (13.7 g, 182 mmol) in DMF (60 mL) at 0 °C was added t -BuOK (20.5 g, 182 mmol) and the mixture was stirred at 0 °C for 1 h. Then a solution of 4-bromo-2-fluorobenzonitrile (12.2 g, 60.8 mmol) in DMF (30 mL) was added and the mixture was stirred at rt overnight. The mixture was partitioned between EtOAc (500 mL) and water (200 mL), the layers were separated and the organic layer was washed with water (2 times), brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 30/1) to give the title compound (8.1 g, 63%) as a white solid. LCMS-D: _sp 2.34 min; m/z 213.0/215.0 [M+H] ⁺ .

xxxv) 4-Methoxy-6-(pyridin-2-yl)benzo[ d ]isoxazol-3-amine I77

a) 2-Fluoro-6-methoxy-4-(pyridin-2-yl)benzonitrile I76

A mixture of 4-bromo-2-fluoro-6-methoxybenzonitrile I5 (244 mg, 1.06 mmol), pyridin _- 2-ylboronic acid (195 mg, 1.59 mmol), CuCl (105 mg, 1.06 mmol), Pd(OAc) ₂ (24 mg, 0.106 mmol), XPhos (100 mg, 0.212 mmol) and _Cs2CO3 (1.38 g, 4.24 mmol) in DMF (10.6 mL) was heated under nitrogen at 100 °C in a sealed 20 mL tube for 16 h. The reaction was repeated three more times on the same scale and the four reactions were quenched with saturated aqueous _NH4Cl , combined and extracted with EtOAc (80 mL × 3). The combined organic extracts were washed with water, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=10/1 to 5/1) to give the title compound (190 mg, 20%) as a yellow solid. LCMS-C: _sp 2.13 min; m/z 229.0 [M+H] ⁺ .

b) 4-Methoxy-6-(pyridin-2-yl)benzo[d]isoxazol-3-amine I77

To a solution of acetohydroxamic acid (178 mg, 2.37 mmol) in anhydrous DMF (20 mL) at 0 °C was added potassium tert -butoxide (266 mg, 2.37 mmol) and the mixture was stirred at 0 °C for 1 h. Then 2-fluoro-6-methoxy-4-(pyridin-2-yl)benzonitrile I76 (180 mg, 0.79 mmol) was added and the mixture was heated at 40 °C overnight. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=200/1 to 100/1 to 60/1) to give the title compound (70 mg, 37%) as a yellow solid. LCMS-C: _sp 0.52 min; m/z 242.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO -d ₆ ) δ 8.73-8.68 (m, 1H), 8.14-8.09 (m, 1H), 7.95-7.88 (m, 1H), 7.70 (d, J = 1.0 Hz, 1H), 7.46 (s, 1H), 7.44-7.38 (m, 1H), 6.01 (s, 2H), 4.01 (s, 3H).

xxxvi) 4-Methoxy-7-phenylbenzo[ d ]isoxazol-3-amine I80

a) 3-Bromo-2-fluoro-6-methoxybenzonitrile I78

A solution of _Br2 (507 mg, 3.2 mmol) in _CCl4 (4.0 mL) was added to a solution of 2-fluoro-6-methoxybenzonitrile (480 mg, 3.2 mmol) and Fe (8.0 mg, 0.1 mmol) in _CCl4 ( _4.0 mL) at -10 °C over a period of 30 min, after which the mixture was allowed to warm to rt and stirred overnight. The mixture was partitioned between water and EtOAc, the layers were separated and the organic layer was washed with saturated aqueous _Na2SO3 ( ₂ times), brine, dried over anhydrous _Na2SO4 , filtered and concentrated to give the title compound (580 mg, 80%) as a white solid. LCMS-C: _sp 2.12 min; m/z 229.9 [M+H] ⁺ .

b) 2-Fluoro-4-methoxy-[1,1'-biphenyl]-3-carbonitrile I79

To a solution of 3-bromo-2-fluoro-6-methoxybenzonitrile I78 (600 mg, 2.6 mmol), _{phenylboronic} acid (636 mg, 5.2 mmol) and _Na2CO3 (829 mg, 7.8 mmol) in 1,4-dioxane (40 mL) and water (10 mL) under _N2 was added Pd( _PPh3 ) ₄ (300 mg, 0.26 mmol) and the mixture was heated at 100°C overnight. The mixture was partitioned between water and EtOAc, the layers were separated and the organic layer was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=10/1 to 3/1) to give the title compound (538 mg, 90%) as a white solid. LCMS-C: _sp 2.43 min; m/z 228.0 [M+H] ⁺ .

c) 4-Methoxy-7-phenylbenzo[d]isoxazol-3-amine I80

To a solution of acetohydroxamic acid (533 mg, 7.11 mmol) in anhydrous DMF (30 mL) at rt was added potassium tert -butoxide (797 mg, 7.11 mmol) and the mixture was stirred at rt for 1 h. Then 2-fluoro-4-methoxy-[1,1'-biphenyl]-3-carbonitrile I79 (538 mg, 2.37 mmol) was added and the mixture was heated at 60 °C overnight. Water was added and the mixture was extracted with EtOAc (30 mL × 3). The combined organic extracts were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=10/1 to 5/1) to give the title compound (413 mg, 72%) as an orange solid. LCMS-C: _sp 1.33 min; m/z 209.0 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.84-7.79 (m, 2H), 7.61 (d, J = 8.1 Hz, 1H), 7.46 (t, J = 7.7 Hz, 2H), 7.38-7.32 (m, 1H), 6.67 (d, J = 8.2 Hz, 1H), 4.00 (s, 3H).

xxxvii) 4-Methoxy-7-(1-methyl-1 H- pyrazol-4-yl)benzo[ d ]isoxazol-3-amine I82

a) 2-Fluoro-6-methoxy-3-(1-methyl-1H-pyrazol-4-yl)benzonitrile I81

To a solution of 3-bromo-2-fluoro-6-methoxybenzonitrile I78 (720 mg, 3.31 mmol), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H - pyrazole (1.30 g, 6.26 _mmol ) and _Na2CO3 (995 mg, 9.39 mmol) in 1,4-dioxane (50 mL) and water (10 mL) under _N2 was added Pd( _PPh3 ) ₄ (358 mg, 0.30 mmol) and the mixture was heated at 100°C overnight. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=100/1) to give the title compound (485 mg, 63%) as a white solid. LCMS-C: _sp 1.26 min; m/z 232.0 [M+H] ⁺ .

b) 4-Methoxy-7-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-amine I82

To a solution of acetohydroxamic acid (474 mg, 6.24 mmol) in anhydrous DMF (20 mL) at rt was added potassium tert -butoxide (700 mg, 6.24 mmol) and the mixture was stirred at rt for 1 h. Then 2-fluoro-6-methoxy-3-(1-methyl- 1H- pyrazol-4-yl)benzonitrile I81 (485 mg, 2.04 mmol) was added and the mixture was heated at 60 °C overnight. Water was added and the mixture was extracted with EtOAc (30 mL × 3). The combined organic extracts were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=50/1) to give the title compound (225 mg, 45%) as a yellow solid. LCMS-C: _sp 0.46 min; m/z 245.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.19 (s, 1H), 7.95 (s, 1H), 7.69 (d, J =8.2 Hz, 1H), 6.74 (d, J =8.2 Hz, 1H), 6.00 (s, 2H), 3.91 (s, 3H), 3.90 (s, 3H).

xxxviii) 5-Chloro-4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I83

To a solution of 4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I9 (300 mg, 1.4 mmol) in DMF (10 mL) was added NCS (192 mg, 1.4 mmol), and the mixture was heated at 50 °C for 2 h. The mixture was diluted with EtOAc (100 mL) and washed with H ₂ O (40 mL × 3). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=7/1 to 5/1) to give the title compound (190 mg, 54%) as a white solid. LCMS-C: _sp 1.21 min; m/z 242.9 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.32 (s, 1H), 6.19 (s, 2H), 4.55 (s, 2H), 3.93 (s, 3H), 3.41 (s, 3H).

xxxix) 4-Methoxy-6-(oxazol-2-yl)benzo[d]isoxazol-3-amine I86

a) 2-(Tributylstannyl)oxazole I84

To a solution of oxazole (500 mg, 7.25 mmol) in THF (15 mL) at -78 °C under N ₂ was added n- BuLi (2.5 M solution in hexanes, 2.9 mL, 7.32 mmol) dropwise and the mixture was stirred at -78 °C for 30 min. Tributyl chlorostannane (1.96 mL, 7.25 mmol) was then added and the mixture was allowed to warm to rt and stirred for 1 h. The solvent was removed under reduced pressure and the residue was taken up in hexanes (50 mL). The resulting precipitate was removed by filtration and the filtrate was concentrated under reduced pressure to give the title compound (2.0 g, 77%) as a colorless oil. ¹ H NMR (400 MHz, DMSO - _d6 ) δ 8.20 (s, 1H), 7.20 (s, 1H), 1.59-1.49 (m, 6H), 1.31-1.26 (m, 6H), 1.16-1.10 (m, 6H), 0.83 (t, J = 7.3 Hz, 9H).

b) 2-Fluoro-6-methoxy-4-(oxazol-2-yl)benzonitrile I85

To a solution of 4-bromo-2-fluoro-6-methoxybenzonitrile I5 (305 mg, 1.33 mmol) in 1,4-dioxane (25 mL) were added 2-(tributylstannyl)oxazole I84 (1.43 g, 3.98 mmol) and Pd( _PPh3 ) ₄ (154 mg, 0.133 mmol), and the mixture was heated at 90°C overnight. The mixture was diluted with water and extracted with EtOAc. The combined organic extracts were washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated. The residue was purified by column chromatography (pet. ether/EtOAc= 8/1) to give the title compound (370 mg, 96%) as a white solid. LCMS-C: _sp 1.86 min; m/z 218.9 [M+H] ⁺ .

c) 4-Methoxy-6-(oxazol-2-yl)benzo[d]isoxazol-3-amine I86

To a solution of acetohydroxamic acid (382 mg, 5.09 mmol) in DMF (25 mL) at 0 °C was added potassium tert -butoxide (570 mg, 5.09 mmol) and the mixture was stirred at rt for 1 h. Then 2-fluoro-6-methoxy-4-(oxazol-2-yl)benzonitrile I85 (370 mg, 1.7 mmol) was added and the mixture was heated at 60 °C for 2 h. The mixture was diluted with EtOAc and washed with water, brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to give the title compound (100 mg, 26%) as a yellow solid. LCMS-C: _sp 0.57 min; m/z 232.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.29 (s, 1H), 7.56 (s, 1H), 7.47-7.42 (m, 1H), 7.27 (s, 1H), 6.09 (s, 2H), 4.00 (s, 3H).

xl) 6-(Oxazol-2-yl)benzo[ d ]isoxazol-3-amine I90

a) 4-Cyano-3-fluorobenzoyl chloride I87

To a solution of 4-cyano-3-fluorobenzoic acid (1.0 g, 6.1 mmol) in DCM (20 mL) at 0 °C was added DMF (0.1 mL) and oxalyl chloride (1.86 g, 12.1 mmol) dropwise and the mixture was stirred at rt overnight. The solvent was removed under reduced pressure to give the title compound (1.2 g) as a white solid, which was used directly in the next step without further purification.

b) 4-Cyano-N-(2,2-dimethoxyethyl)-3-fluorobenzamide I88

To a solution of 4-cyano-3-fluorobenzoyl chloride I87 (1.1 g, 6.06 mmol) and _Et3N (1.84 g, 18 mmol) in DCM (20 mL) was added 2,2-dimethoxyethanamine (955 mg, 9.1 mmol) at 0 °C, and the mixture was stirred for 2 h. The organic extract was washed with water and brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=1/1, v/v) to give the title compound (1.2 g, 78%) as a white solid. LCMS-D: sp _. 1.55 min, m/z 274.9 [M+Na] ⁺ .

c) 3-Amino-N-(2,2-dimethoxyethyl)benzo[d]isoxazole-6-carboxamide I89

To a solution of acetohydroxamic acid (448 mg, 5.95 mmol) in DMF (30 mL) at 0 °C was added t -BuOK (889 mg, 7.92 mmol) portionwise and the mixture was stirred for 1 h. Then 4-cyano- N- (2,2-dimethoxyethyl)-3-fluorobenzamide I88 (500 mg, 1.98 mmol) was added and the mixture was heated at 40 °C overnight. The mixture was poured into water and extracted with EtOAc. The organic layer was washed with water and brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=1/1, v/v) to give the title compound (380 mg, 72%) as a yellow solid. LCMS-D: _sp 0.59 min, m/z 287.9 [M+Na] ⁺ .

d) 6-(Oxazol-2-yl)benzo[d]isoxazol-3-amine I90

A mixture of 3-amino- N- (2,2-dimethoxyethyl)benzo[ d ]isoxazole-6-carboxamide I89 (240 mg, 0.9 mmol) and _P2O5 (193 mg, 1.36 mmol) _in methanesulfonic acid (10 mL) was heated at 150 °C under microwave irradiation for 30 min. The mixture was poured into water, basified with aqueous KOH solution and extracted with EtOAc. The organic extract was washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=10/1, v/v) to give the title compound (50 mg, 28%) as a solid. LCMS-D: ep. _beat 1.57 min, m/z 201.9 [M+H] ⁺ .

xli) 5-Ethyl-1-methyl-2-oxo-1,2-dihydropyridine-3-sulfonyl chloride I94

a) 5-Ethyl-2-methoxypyridine I91

To a solution of 5-bromo-2-methoxypyridine (10.2 g, 54.25 mmol) in THF (200 mL) at -78 °C under N ₂ was added n- BuLi (2.5 M solution in hexane, 24.0 mL, 60.0 mmol) dropwise and the mixture was stirred at -78 °C for 1.5 h. Iodoethane (12.7 g, 81.4 mmol) was then added dropwise and the mixture was stirred at -78 °C for 20 min, then warmed to rt and stirred for 30 min. The reaction was quenched with water (5 mL) and the solvent was removed under reduced pressure. The residue was dissolved in DCM, washed with water, brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 5/1) to give the title compound (2.8 g, 38%) as a colorless oil. LCMS-D: _sp 1.80 min; m/z 138.1 [M+H] ⁺ .

b) 5-Ethylpyridin-2(1H)-one I92

A solution of 5-ethyl-2-methoxypyridine I91 (1.6 g, 11.66 mmol) in conc. HCl (30 mL) was heated at 100 °C overnight. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography (DCM/MeOH=20/1) to give the title compound (800 mg, 56%) as a white solid, which was used directly in the next step.

c) 5-Ethyl-1-methylpyridin-2(1H)-one I93

A mixture of 5-ethylpyridin-2( 1H )-one I92 (800 mg, 6.5 mmol), _{K2CO3 (} 1.8 g, 13 mmol) and iodomethane (1.85 g, 13 mmol) in MeOH (20 mL) was heated at 50 °C under _N2 overnight. The solvent was removed under reduced pressure, the residue was dissolved in DCM (100 mL), washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=40/1) to give the title compound (500 mg, 56%) as a colorless oil. LCMS-D: _sp 0.68 min; m/z 138.1 [M+H] ⁺ .

d) 5-Ethyl-1-methyl-2-oxo-1,2-dihydropyridine-3-sulfonyl chloride I94

A mixture of chlorosulfonic acid (6 mL) and 5-ethyl-1-methylpyridin-2( 1H )-one I93 (0.6 g, 4.37 mmol) was heated at 150 °C under N ₂ for 3 h, then allowed to cool to rt and poured onto ice (100 g). The mixture was extracted with DCM (50 mL x 3), the combined organic extracts were washed twice with ice water, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH = 100/1 to 20/1) to give the title compound (200 mg, 12%) as a yellow solid. ¹ H NMR (400 MHz, DMSO _-d6 ) δ 7.87-7.84 (m, 1H), 7.71 (s, 1H), 3.47 (s, 3H), 2.39 (k, J= 7.6 Hz, 2H), 1.09 (t, J= 7.5 Hz, 3H). LCMS-D: vr. _beat 1.58 min; m/z 236.0 [M+H] ⁺ .

xlii) 5-Bromo-2,3-dihydrobenzofuran-7-sulfonyl chloride I95

5-Bromo-2,3-dihydrobenzofuran (2.0 g, 10 mmol) was slowly added to chlorosulfonic acid (6 mL) at -5 °C, and the mixture was stirred at -5 °C for 30 min. The mixture was poured into ice water (100 mL) and extracted with EtOAc (180 mL × 2). The combined organic extracts were washed with water (250 mL × 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=15/1) to give the title compound (1.45 g, 48%) as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.80-7.76 (m, 1H), 7.63-7.58 (m, 1H), 4.91 (m, 2H), 3.35 (m, 2H). LCMS-D: vr. _beat 2.74 min; m/z 318.8/320.8 [M+Na] ⁺ .

xliii) 4,6-Dimethoxy-2,3-dihydro-1 H -indene-5-sulfonyl chloride I98

a) 5,7-Dimethoxy-2,3-dihydro-1H-inden-1-one I96

A mixture of 3-(3,5-dimethoxyphenyl)propanoic acid (5 g, 23.8 mmol) and methanesulfonic acid (24 mL) was heated at 90 °C for 10 min, cooled to rt and poured into water. The pH of the mixture was adjusted to 9 with 10 M aq KOH and extracted with EtOAc (5 times). The combined organic extracts were washed with water, brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (3.5 g, 76%) as a white solid. LCMS-D: _sp 1.76 min; m/z 193.1 [M+H] ⁺ .

b) 4,6-Dimethoxy-2,3-dihydro-1H-indene I97

A mixture of 5,7-dimethoxy-2,3-dihydro- 1H -inden-1-one I96 (3.0 g, 15.6 mmol) and triethylsilane (7.3 g, 62.4 mmol) in TFA (20 mL) was stirred at rt under _N2 for 11 h. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=20/1) to give the title compound (2.0 g, 72%) as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 6.42 (s, 1H), 6.29 (d, J= 2.0 Hz, 1H), 3.81 (s, 3H), 3.80 (s, 3H), 2.89 (t, J= 7.5 Hz, 2H), 2.80 (t, J= 7.3 Hz, 2H), 2.13-2.02 (m, 2H).

c) 4,6-Dimethoxy-2,3-dihydro-1H-indene-5-sulfonyl chloride I98

To a solution of 4,6-dimethoxy-2,3-dihydro- 1H- indene I97 (1 g, 5.6 mmol) and TMEDA (0.72 g, 6.17 mmol) in n- hexane (20 mL) at -70 °C was added n- BuLi (2.5 M in hexane, 2.5 mL, 6.17 mmol) dropwise and the mixture was allowed to warm to 0 °C and stirred for 2 h. The mixture was recooled to -65 °C, bubbled with SO ₂ gas for 20 min and then allowed to warm slowly to 10 °C. The resulting precipitate was collected by filtration and washed with dry diethyl ether. The solid was suspended in n -hexane (20 mL), cooled to 0 °C, and _SO2Cl2 (0.83 g, 6.2 mmol) was added dropwise. The mixture was stirred at 0 °C under _N2 for 1 h and then filtered. _The filter cake was dissolved in diethyl ether and washed with water, brine, dried over _{anhydrous Na2SO4} , filtered, and concentrated under reduced pressure to give the title compound (550 mg, 35%) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.72 (s, 1H), 3.95 (s, 3H), 3.94 (s, 3H), 2.96 (q, J= 7.6 Hz, 4H), 2.18-2.08 (m, 2H).

xliv) 2-Methoxy-5-phenoxybenzenesulfonyl chloride I100

a) 2-Methoxy-5-phenoxybenzenesulfonic acid I99

To a solution of 1-methoxy-4-phenoxybenzene (2 g, 10 mmol) in DCM (15 mL) at 0 °C was added dropwise a solution of chlorosulfonic acid (0.35 mL) in DCM (10 mL), and the mixture was stirred at 0 °C for 15 min. The mixture was slowly poured into ice water (100 mL), and then concentrated under reduced pressure. The residue was washed with DCM (100 mL × 2) and dried to give the title compound (560 mg, 40%) as an off-white solid. LCMS-D: _sp 0.62 min; m/z 281.0 [M+H] ⁺ .

b) 2-Methoxy-5-phenoxybenzenesulfonyl chloride I100

A mixture of 2-methoxy-5-phenoxybenzenesulfonic acid I99 (250 mg, 0.9 mmol) and PCl ₅ (284 mg, 1.3 mmol) in POCl ₃ (3 mL) was heated at 90 °C under N ₂ for 1 h. The mixture was then slowly added to ice water (20 mL) and extracted with DCM (15 mL x 2). The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (51 mg, 18%) as a yellow oil. LCMS-D: _sp 2.72 min; m/z 295.0 [M-Cl+OCH ₃ ] ⁺ , 317.0 [M-Cl+OCH ₃ +Na] ⁺ .

xlv) 2,6-Dimethoxy-3-(trifluoromethyl)benzenesulfonyl chloride I102

a) 2,4-Dimethoxy-1-(trifluoromethyl)benzene I101

To a mixture _{of (2,4-dimethoxyphenyl)boronic acid (3.0 g, 16.5 mmol), CF3SO2Na (} 18.0 g, 115.4 mmol), Cu(OAc) ₂ (748 mg, 4.1 mmol), imidazole (281 mg, 4.1 mmol), 2,4,6-collidine (3.0 g, 33.0 mmol) and _NH4Cl (11.3 g, 206.0 mmol) in water (16.5 mL) and DCM (100 mL) at 0 °C was added dropwise t -BuOOH (3.6 mL, 4.1 mmol) and the mixture was stirred at rt for 16 h. After that, the layers were separated and the aqueous layer was extracted with DCM. The combined organic layers were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether) to give the title compound (900 mg, 27%) as an oil. ¹ H NMR (400 MHz, _CDCl3 ) δ 7.42-7.36 (m, 1H), 6.45-6.37 (m, 2H), 3.79 (s, 3H), 3.76 (s, 3H).

b) 2,6-Dimethoxy-3-(trifluoromethyl)benzenesulfonyl chloride I102

To a solution of 2,4-dimethoxy-1-(trifluoromethyl)benzene I101 (1.5 g, 7.3 mmol) and TMEDA (0.93 g, 8.0 mmol) in n- hexane (30 mL) at -78 °C under N ₂ was added dropwise n -BuLi (2.5 M in hexane, 3.2 mL, 8.0 mmol) and the mixture was stirred at 0 °C for 1 h. The mixture was bubbled with SO ₂ gas at -78 °C for 20 min, then allowed to warm to 0 °C and stirred for 1 h. The resulting precipitate was collected by filtration and washed with hexane. The filter cake was suspended in n -hexane (30 mL), cooled to 0 °C and _{SO2Cl2 (} 1.1 g, 8.0 mmol) was added dropwise. The mixture was stirred at 0 °C for 1 h, the solids were collected by filtration and washed with cold n -hexane. The filter cake was dissolved in ether and washed with water. The aqueous phase was extracted with ether, the combined organic layers were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=5/1) to give the title compound (1.5 g, 68%) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.90–7.84 (m, 1H), 6.98–6.92 (m, 1H), 4.09 (s, 3H), 4.04 (s, 3H).

xlvi) 3-Ethyl-2,6-dimethoxybenzenesulfonyl chloride I106

a) 2,4-Dimethoxy-1-vinylbenzene I103

A suspension of 1-bromo-2,4-dimethoxybenzene (4.0 g, 18.4 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (3.4 g, 22.1 mmol), Pd(dppf) _Cl2⋅DCM (753 mg, 0.92 mmol), and _K2CO3 (7.6 g, 55.2 mmol) _in 1,4-dioxane (40 mL) and water (10 mL) was heated at 90 °C under _N2 overnight. The mixture was filtered through a pad of celite and washed with EtOAc. The filtrate was diluted with water and extracted with EtOAc (60 mL × 3). The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether) to give the title compound (2.6 g, 86%) as a yellow oil. LCMS-D: _sp 2.28 min; m/z 165.0 [M+H] ⁺ .

b) Ethyl-2,4-dimethoxybenzene I104

To a solution of 2,4-dimethoxy-1-vinylbenzene I103 (2.6 g, 15.8 mmol) in EtOAc (50 mL) was added 10% Pd/C (300 mg) and the mixture was stirred under _H2 at rt overnight. The catalyst was removed by filtration through celite and washed with EtOAc. The filtrate was concentrated under reduced pressure to give the title compound (2.0 g, 83%) as a yellow oil. LCMS-D: sp _temp 2.39 min; m/z 167.1 [M+H] ⁺ .

c) 3-Ethyl-2,6-dimethoxybenzenesulfonic acid I105

Prepared from ethyl 2,4-dimethoxybenzene I104 according to the procedure described for 2,6-dimethoxybenzenesulfonyl chloride I111. The resulting product was found to be mainly 3-ethyl-2,6-dimethoxybenzenesulfonic acid. LCMS-D: _sp 2.36 min; m/z 247.0 [M+H] ⁺ .

d) 3-Ethyl-2,6-dimethoxybenzenesulfonyl chloride I106

A mixture of 3-ethyl-2,6-dimethoxybenzenesulfonic acid I105 (300 mg, 1.22 mmol) and thionyl chloride (6 mL) was heated at 95 °C for 3 h and then concentrated under reduced pressure to give the title compound (322 mg, 100%) as a brown oil, which was used directly in the next step.

xlvii) 7-Methoxyquinoline-8-sulfonyl chloride I107

Chlorosulfonic acid (1.8 g, 15.7 mmol) was added dropwise to 7-methoxyquinoline (500 mg, 3.14 mmol) at 0 °C and the mixture was heated at 100 °C for 1 h. The mixture was allowed to cool to rt, poured onto ice and then neutralized with saturated aqueous NaHCO ₃ solution. The mixture was extracted with EtOAc (30 mL x 3), the combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (280 mg, 34%) as a yellow solid. LCMS-D: _sp 0.27 min; m/z 239.9 [M-Cl+ _H2O ] ⁺

xlviii) 6-Methoxy-2,3-dihydro-1 H -indene-5-sulfonyl chloride I108

To a solution of 5-methoxy-2,3-dihydro- 1H -indene (3.1 g, 20.9 mmol) in DCM (40 mL) at -5 °C under N ₂ atmosphere was added dropwise chlorosulfonic acid (6.5 g, 62.8 mmol), and the mixture was stirred at -5 °C for 40 min. The reaction was quenched with ice water (20 mL), and the mixture was extracted with EtOAc (30 mL × 2). The combined organic extracts were washed with water (50 mL × 3), dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=20/1) to give the title compound (3.1 g, 60%) as a white solid, which was used directly in the next step. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.77 (d, J= 1.1 Hz, 1H), 6.98 (s, 1H), 4.02 (s, 3H), 2.99 (t, J= 7.5 Hz, 2H), 2.91 (t, J= 7.4 Hz, 2H), 2.19-2.10 (m, 2H).

xlix) 3-Methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonyl chloride I109

Prepared from 6-methoxy-1,2,3,4-tetrahydronaphthalene and chlorosulfonic acid according to the procedure described for 6-methoxy-2,3-dihydro- 1H -indene-5-sulfonyl chloride I108 and used directly in the next step without purification.

l) 4-Bromo-2-methoxybenzenesulfonyl chloride I110

To chlorosulfonic acid (16 mL) at -5 °C was slowly added 1-bromo-3-methoxybenzene (15.0 g, 80 mmol), and the mixture was stirred at -5 °C for 5 min. The mixture was poured into ice water (50 mL) and extracted with EtOAc (80 mL × 2). The combined organic extracts were washed with water (150 mL × 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1) to give the title compound (2.6 g, 17%) as a yellow solid, which was used directly in the next step.

li) 2,6-Dimethoxybenzenesulfonyl chloride I111

To a solution of 1,3-dimethoxybenzene (5.0 g, 36 mmol) and TMEDA (4.6 g, 39.8 mmol) in n -hexane (100 mL) at 0 °C under N ₂ was added dropwise n- BuLi (2.5 M solution in hexanes, 16.0 mL, 39.8 mmol) while maintaining the internal reaction temperature below 5 °C. The mixture was stirred at 0 °C for 20 min, then cooled to -78 °C and bubbled with SO ₂ gas for 20 min. The mixture was then allowed to warm slowly to 10 °C, and the resulting precipitate was collected by filtration and washed with dry diethyl ether. The solid was suspended in n -hexane (100 mL), cooled to 0 °C, and a solution of _SO2Cl2 ( _4.9 g, 36 mmol) in n -hexane (20 mL) was added dropwise while maintaining the internal temperature below 3 °C. The mixture was then stirred at 0 °C for 1 h, the solids were collected by filtration and washed with cold n -hexane. The solids were then partitioned between diethyl ether and water, the layers were separated, and the aqueous layer was further extracted with _diethyl ether. The combined organic extracts were dried over anhydrous _Na2SO4 , filtered, and concentrated under reduced pressure to give the title compound (4.0 g, 47%) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.54 (t, J= 8.4 Hz, 1H), 6.66 (d, J= 8.4 Hz, 2H), 3.97 (s, 6H).

lii) 5-Ethyl-2-methoxybenzenesulfonyl chloride I112

1-Ethyl-4-methoxybenzene (5.0 g, 37 mmol) was added dropwise to chlorosulfonic acid (20 mL) at 0 °C, the mixture was stirred at rt for 2 h, then poured onto ice and extracted with EtOAc (50 mL x 3). The combined organic extracts were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc = 100/1 to 30/1) to give the title compound (4.6 g, 53%) as a white solid. LCMS-D: _sp 2.70 min; m/z 256.9 [M+Na] ⁺ .

liii) 2,4-Dimethoxy-[1,1'-biphenyl]-3-sulfonyl chloride I114

a) 2,4-Dimethoxy-1,1'-biphenyl I113

A suspension of 1-bromo-2,4-dimethoxybenzene (5.0 g, 23.0 mmol), phenylboronic acid (3.4 g, 27.6 mmol), Pd( _PPh3 ) ₄ (1.3 g, 1.15 mmol), and potassium carbonate (7.3 g, 69.0 mmol) in 1,4-dioxane (30 mL) and water (6 mL) was heated at 90 °C under _N2 for 16 h. The mixture was filtered through a pad of celite and washed with EtOAc. The filtrate was diluted with water and extracted with _EtOAc (30 mL×3). The combined organic extracts were dried over _Na2SO4 , filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 10/1) to give the title compound (2.8 g, 57%) as a yellow oil. LCMS-D: _sp 2.46 min; m/z 215.0 [M+H] ⁺ .

b) 2,4-Dimethoxy-[1,1'-biphenyl]-3-sulfonyl chloride I114

To a solution of 2,4-dimethoxy-1,1'-biphenyl I113 (1.0 g, 4.70 mmol) and TMEDA (601 mg, 5.20 mmol) in n- hexane (40 mL) at 0 °C under N ₂ was added dropwise n- BuLi (2.5 M solution in hexanes, 2.1 mL, 5.20 mmol) while maintaining the internal reaction temperature below 5 °C. The mixture was stirred at 0 °C for 20 min, then cooled to -70 °C and bubbled with SO ₂ gas for 20 min. The mixture was then allowed to warm slowly to 10 °C, the resulting precipitate was collected by filtration and washed with dry diethyl ether. The solid was suspended in n -hexane (40 mL), cooled to 0 °C, and a solution of _SO2Cl2 ( ₆₃₄ mg, 4.7 mmol) in n -hexane (5 mL) was added dropwise while maintaining the internal temperature below 3 °C. The mixture was then stirred at 0 °C for 1 h, the solids were collected by filtration and washed with cold n -hexane. The solids were then partitioned between diethyl ether and water, the layers were separated, and the aqueous layer was further extracted with _diethyl ether. The combined organic extracts were dried over _Na2SO4 , filtered, and concentrated under reduced pressure to give the title compound (590 mg, 40%) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.48-7.35 (m, 4H), 7.34-7.21 (m, 2H), 6.87 (m, 1H), 3.76 (s, 3H), 3.29 (s, 3H).

liv) 3,5-Dimethoxy-[1,1'-biphenyl]-4-sulfonyl chloride I116

a) 3,5-Dimethoxy-1,1'-biphenyl I115

A suspension of 1-bromo-3,5-dimethoxybenzene (5.0 g, 23.0 mmol), phenylboronic acid (2.8 g, 23.0 mmol), Pd(dppf)Cl ₂ (0.57 g, 0.69 mmol) and potassium carbonate (4.8 g, 34.6 mmol) in 1,4-dioxane (80 mL) and water (20 mL) was heated at 90 °C under N ₂ for 4 h. The mixture was diluted with water, extracted with EtOAc, the combined organic extracts were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=500/1 to 200/1 to 100/1) to give the title compound (5.2 g, 100%) as a white solid. LCMS-C: _sp 2.47 min; m/z 215.0 [M+H] ⁺ .

b) 3,5-Dimethoxy-[1,1'-biphenyl]-4-sulfonyl chloride I116

Prepared from 3,5-dimethoxy-1,1'-biphenyl I115 according to the procedure described for 2,4-dimethoxy-[1,1'-biphenyl]-3-sulfonyl chloride I114, ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.62-7.55 (m, 2H), 7.54-7.44 (m, 3H), 6.81 (s, 2H), 4.04 (s, 6H).

lv) 4-Methoxy-6-((2,2,2-trifluoroethoxy)methyl)benzo[ d ]isoxazol-3-amine I118

a) 2-Fluoro-6-methoxy-4-((2,2,2-trifluoroethoxy)methyl)benzonitrile I117

To a solution of 2-fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7 (500 mg, 2.76 mmol) in dry THF (50 mL) at 0 °C under N ₂ was added NaH (60 wt % dispersion in oil, 331 mg, 8.28 mmol) followed by 2,2,2-trifluoroethyl trifluoromethanesulfonate (1.9 g, 8.28 mmol) and the mixture was stirred at 0 °C for 1 h, then allowed to warm to rt and stirred overnight. The mixture was diluted with water and extracted with EtOAc. The organic extract was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The reaction was repeated twice using 2-fluoro-4-(hydroxymethyl)-6-methoxybenzonitrile I7 (100 mg, 0.55 mmol), the three batches were combined and purified by column chromatography (pet. ether/EtOAc=5/1 to 2/1) to give the title compound (577 mg, 57%) as a white solid. LCMS-C: _sp 2.43 min; m/z 263.9 [M+H] ⁺ .

b) 4-Methoxy-6-((2,2,2-trifluoroethoxy)methyl)benzo[d]isoxazol-3-amine I118

A suspension of acetohydroxamic acid (86 mg, 1.14 mmol) and t -BuOK (128 mg, 1.14 mmol) in anhydrous DMF (10 mL) was stirred at rt for 1 h. Then 2-fluoro-6-methoxy-4-((2,2,2-trifluoroethoxy)methyl)benzonitrile I117 (100 mg, 0.38 mmol) was added and the mixture was stirred at rt overnight. The mixture was diluted with water and extracted with EtOAc. The organic extract was washed with brine, dried over _Na2SO4 , _filtered and concentrated under reduced pressure. The reaction was scaled up accordingly using 2-fluoro-6-methoxy-4-((2,2,2-trifluoroethoxy)methyl)benzonitrile I117 (400 mg, 1.52 mmol), the two batches were combined and purified by column chromatography (pet. ether/EtOAc=20/1 to 5/1) to give the title compound (350 mg, 67%) as a yellow solid. LCMS-C: _sp 2.08 min; m/z 277.0 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 6.96 (s, 1H), 6.68 (s, 1H), 5.94 (s, 2H), 4.74 (s, 2H), 4.13 (k, J =9.4 Hz, 2H), 3.90 (s, 3H).

lvi) 6-(Difluoromethoxy)-4-methoxybenzo[ d ]isoxazol-3-amine I121

a) 4-(Difluoromethoxy)-2,6-difluorobenzonitrile I119

To a suspension of KOH (22.0 g, 392 mmol) in acetonitrile (30 mL) and water (30 mL) at -20 °C was added 2,6-difluoro-4-hydroxybenzonitrile (3.1 g, 20.0 mmol) portionwise, followed by diethyl (bromodifluoromethyl)phosphonate (10.0 g, 37.4 mmol), and the mixture was stirred at rt overnight. Water was added and the mixture was extracted with EtOAc (50 mL x 3). The combined organic extracts were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (4.0 g, 97%) as a colorless oil. LCMS-C: _sp 2.11 min; m/z 205.9 [M+H] ⁺ .

b) 4-(Difluoromethoxy)-2-fluoro-6-methoxybenzonitrile I120

To a solution of 4-(difluoromethoxy)-2,6-difluorobenzonitrile I119 (2.52 g, 12.3 mmol) in dry THF (30 mL) was added NaOMe (1.32 g, 24.57 mmol) portionwise and the mixture was heated at 40 °C overnight. Water was added and the mixture was extracted with EtOAc (30 mL x 3). The combined organic extracts were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc = 10/1 to 5/1) to give the title compound (663 mg, 25%) as a white solid. LCMS-C: _sp 2.11 min; m/z 217.9 [M+H] ⁺ .

c) 6-(Difluoromethoxy)-4-methoxybenzo[d]isoxazol-3-amine I121

A suspension of acetohydroxamic acid (680 mg, 9.15 mmol) and t- BuOK (1.03 g, 9.15 mmol) in anhydrous DMF (50 mL) was stirred at rt for 1 h. Then 5-(difluoromethoxy)-1-fluoro-2-isocyano-3-methoxybenzene I120 (663 mg, 3.05 mmol) was added and the mixture was stirred at rt overnight. The mixture was diluted with water and extracted with EtOAc (30 mL × 3). The combined organic extracts were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=10/1 to 8/1) to give the title compound (186 mg, 26%) as a light orange solid. LCMS-C: _sp 1.14 min; m/z 231.0 [M+H] ⁺ .

lviii) 5-Methyl-6-(pyridin-2-yl)benzo[ d ]isoxazol-3-amine I124

a) 2-Fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile I122

A mixture of 4-bromo-2-fluoro-5-methylbenzonitrile (500 mg, 2.34 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (1.78 g, 2.34 mmol), potassium acetate (918 mg, 9.36 mmol) and Pd(dppf) _Cl2 (188 mg, 0.23 mmol) in 1,4-dioxane (20 mL) was heated at reflux under _N2 for 3 h. The mixture was diluted with water, extracted with EtOAc (300 mL), the organic layer was washed with water (50 mL × 3), dried _{over Na2SO4} , filtered and concentrated under reduced pressure to give the title compound. the title compound (1.56 g, 88%), which was used in the next step without further purification. LCMS-C: _sp 2.75 min; m/z 262.0 [M+H] ⁺ .

b) 2-Fluoro-5-methyl-4-(pyridin-2-yl)benzonitrile I123

To a solution of 2-fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile I122 (1.32 g, 5.1 mmol) and 2-bromopyridine (1.69 g, 7.65 mmol) in 1,4-dioxane (50 mL) and water (10 mL) under _N2 were added Pd( _PPh3 ) ₄ (589 mg, 0.5 _mmol ) and _Na2CO3 (2.16 g, 20.4 mmol) and the mixture was heated at 100°C for 3 h. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=8/1 to 3/1) to give the title compound (440 mg, 88%) as a red solid. LCMS-C: _sp 1.09 min; m/z 213.0 [M+H] ⁺ .

c) 5-Methyl-6-(pyridin-2-yl)benzo[d]isoxazol-3-amine I124

A suspension of acetohydroxamic acid (255 mg, 3.39 mmol) and t -BuOK (381 mg, 3.39 mmol) in anhydrous DMF (30 mL) was stirred at 0 °C for 1 h. Then 2-fluoro-5-methyl-4-(pyridin-2-yl)benzonitrile I123 (240 mg, 1.13 mmol) was added and the mixture was allowed to warm to rt and stirred overnight. Water was added and the mixture was extracted with EtOAc (50 mL × 3). The combined organic extracts were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=8/1 to 3/1) to give the title compound (180 mg, 73%) as a white solid. LCMS-C: _sp 0.50 min; m/z 226.0 [M+H] ⁺ .

lix) 7-Bromo-5-methylbenzo[ d ]isoxazol-3-amine I129

a) 3-Bromo-2-fluoro-5-methylbenzoic acid I125

To a solution of 2-bromo-1-fluoro-4-methylbenzene (10.0 g, 53 mmol) and diisopropylamine (5.9 g, 58 mmol) in anhydrous THF (200 mL) at -78 °C under N ₂ was added dropwise n -BuLi (2.5 M solution in hexanes, 25.6 mL, 64.0 mmol) and the mixture was stirred at -78 °C for 1 h. Excess solid CO ₂ (dry ice) was added and stirring was continued at -78 °C for 3 h. The mixture was diluted with water (500 mL) and extracted with EtOAc (500 mL). The organic layer was washed with brine, dried over _Na2SO4 , filtered and concentrated under reduced pressure to give the title compound (12.3 g, 100%) as _a brown solid, which was used in the next step without further purification. LCMS-C: _sp 2.03 min; m/z 232.8 [M+H] ⁺ .

b) 3-Bromo-2-fluoro-5-methylbenzoyl chloride I126

To a solution of 3-bromo-2-fluoro-5-methylbenzoic acid I125 (12.3 g, 53 mmol) and DMF (4 drops) in DCM (100 mL) at rt under _N2 was added oxalyl chloride (13.0 g, 106 mmol) dropwise and the mixture was stirred for 2 h. The mixture was concentrated under reduced pressure to give the title compound (14.0 g, 100%) as a brown solid, which was used in the next step without further purification.

c) 3-Bromo-2-fluoro-5-methylbenzamide I127

A solution of 3-bromo-2-fluoro-5-methylbenzoyl chloride I126 (14.0 g, 53 mmol) in DCM (100 mL) was added dropwise to 30% aqueous ammonium hydroxide solution (100 mL) and the mixture was stirred for 2 h. The mixture was diluted with EtOAc (200 mL), washed with water (200 mL × 3) and brine, _the organic layer was dried over _Na2SO4 , filtered and concentrated under reduced pressure to give the title compound (12.0 g, 97%) as a brown solid, which was used in the next step without further purification. LCMS-C: _sp 1.01 min; m/z 231.9 [M+H] ⁺ .

d) 3-Bromo-2-fluoro-5-methylbenzonitrile I128

A solution of 3-bromo-2-fluoro-5-methylbenzamide I127 (10.0 g, 43.0 mmol) and thionyl chloride (15.4 g, 129 mmol) in DMF (100 mL) was heated at 100 °C for 3 h. The mixture was diluted with EtOAc (200 mL), washed with water (400 mL × 5) and brine, the organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (5.0 g, 54%) as a brown solid, which was used in the next step without further purification. LCMS-C: _sp 2.50 min; m/z 213.9 [M+H] ⁺ .

e) 7-Bromo-5-methylbenzo[d]isoxazol-3-amine I129

A suspension of acetohydroxamic acid (5.27 g, 70.2 mmol) and t -BuOK (7.88 g, 70.2 mmol) in anhydrous DMF (200 mL) was stirred at 0 °C for 1 h. Then 3-bromo-2-fluoro-5-methylbenzonitrile I128 (5.0 g, 23.4 mmol) was added, the mixture was allowed to warm to rt and stirred overnight. The mixture was diluted with EtOAc (300 mL), washed with water (600 mL × 4) and brine, the organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=10/1) to give the title compound (2.8 g, 52%) as a yellow solid. LCMS-C: _sp 0.50 min; m/z 226.9 [M+H] ⁺ .

Synthesis of compounds by examples

Examples 1-45 (Table A):

Method AA

To a solution of 4-chlorobenzo[ d ]isoxazol-3-amine (50 mg, 0.297 mmol) in THF (3 mL) was added LiHMDS (1 M in THF, 445 μL, 0.445 mmol) and stirred at room temperature for 10 min. Sulfonyl chloride (0.445 mmol) was added and the reaction was stirred for 16 h at room temperature. The volume of volatiles was reduced to approximately 1 mL, after which DCM (3 mL) and water (3 mL) were added and the mixture was stirred for 10 min. The mixture was passed through a phase separator, then the organic fraction was loaded onto a 1 g amine-modified silica gel cartridge (Biotage), the cartridge was washed with MeOH (6 mL), and the product was eluted with HCl (2 M, 1:1 methanol:1,4-dioxane 6 mL). The HCl washes were then evaporated in vacuo to give the desired product.

Method AB

To a solution of 4-chlorobenzo[ d ]isoxazol-3-amine (50 mg, 0.297 mmol) in THF (3 mL) was added LiHMDS (1 M in THF, 445 μL, 0.445 mmol) and stirred at room temperature for 10 min. Sulfonyl chloride (0.445 mmol) was added and the reaction was stirred for 16 h at room temperature. The volume of volatiles was reduced to approximately 1 mL, after which DCM (3 mL) and water (3 mL) were added and the mixture was stirred for 10 min. The mixture was passed through a phase separator, then the organic fraction was loaded onto a 1 g amine-modified silica gel cartridge (Biotage), the cartridge was washed with MeOH (6 mL), and the product was then eluted with HCl solution (2 M, 1:1 methanol:1,4-dioxane 6 mL). The HCl washings were then evaporated in vacuo to give the crude product, which was loaded onto silica gel and purified by silica gel column chromatography (Biotage Isolera, SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline, 40-60 °C) to give the desired product.

AC Method

To a solution of 4-chlorobenzo[ d ]isoxazol-3-amine (50 mg, 0.297 mmol) in THF (3 mL) was added LiHMDS (1 M in THF, 445 μL, 0.445 mmol) and stirred at room temperature for 10 min. Sulfonyl chloride (0.445 mmol) was added and the reaction was stirred for 16 h at room temperature. The volume of volatiles was reduced to approximately 1 mL, after which DCM (3 mL) and water (3 mL) were added and the mixture was stirred for 10 min. The mixture was passed through a phase separator, then the organic fraction was loaded onto a 1 g amine-modified silica gel cartridge (Biotage), the cartridge was washed with MeOH (10 mL), and the product was eluted with HCl in methanol (~1.25 M, 10 mL). The HCl washes were then evaporated in vacuo to yield the desired product.

AD Method

A solution of 4-chlorobenzo[ d ]isoxazol-3-amine (50 mg, 0.298 mmol) and sulfonyl chloride (2 equiv, 0.595 mmol) in pyridine (1.5 mL) was irradiated in a microwave reactor for 2 h at 100 °C. After cooling, the reaction mixture was loaded onto silica gel and purified by silica gel column chromatography (Biotage Isolera, 24 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline, 40-60 °C) to afford the desired product.

AE Method

A suspension of sulfonyl chloride (2 equiv, 1.19 mmol) and 4-chlorobenzo[ d ]isoxazol-3-amine (100 mg, 0.593 mmol) in pyridine (1.5 mL) was irradiated in a microwave reactor for 2 h at 100 °C. After cooling, the mixture was loaded onto silica gel and purified by silica gel column chromatography (Biotage Isolera, 24 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline, 40-60 °C, then 0-40% MeOH in EtOAc) to afford the desired product.

AF Method

A suspension of sulfonyl chloride (2 equiv, 1.19 mmol) and 4-chlorobenzo[ d ]isoxazol-3-amine (100 mg, 0.593 mmol) in pyridine (1 mL) was irradiated in a microwave reactor for 1 h at 80 °C. After cooling, the mixture was loaded onto silica gel and purified by silica gel column chromatography (Biotage Isolera, 24 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline, 40-60 °C) to afford the desired product.

Table A

Structure Name Analytical data Method 1 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-(methoxymethyl)benzenesulfonamide ZHMS-A : vr. beat 6.538 min, m/z 352.1 [MH] ^- Method AB 2 N- (4-chlorobenzo
[ d ]isoxazol-3-yl)-3-(methoxymethyl)
benzenesulfonamide ZHMS-A: vr. beat 6.374 min, m/z 353.1 [M+H] ⁺ Method AA 3 3,4-dichloro- N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-methoxy benzenesulfonamide ZHMS-A: vr. beat 6.751 min, m/z 409.0 [M+H] ⁺ Method AA 4 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-phenoxybenzene sulfonamide ZHMS-A: vr. beat 6.745 min, m/z 401.1 [M+H] ⁺ Method AA 5 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-5-fluoro-2-methoxybenzene sulfonamide ZHMS-A: vr. beat 6.360 min, m/z 357.1 [M+H] ⁺ Method AA 6 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-methyl-2 H -benzo[ d ][1,2,3]triazole-4-sulfonamide ZHMS-A: vr. beat 6,200 min, m/z 364.1 [M+H] ⁺ Method AA 7 N- (4-chlorobenzo
[ d ]isoxazol-3-yl)benzo[ c ][1,2,5]thiadiazole-4-sulfonamide ZHMS-A: vr. beat 6.346 min, m/z 367.0 [M+H] ⁺ Method AA 8 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-hydroxybenzo[ d ]oxazole-6-sulfonamide ZHMS-A: vr. beat 5.856 min, m/z 364.0 [MH] ^- Method AA 9 4-(benzyloxy) -N- (4-chlorobenzo[ d ]isoxazol-3-yl)benzenesulfonamide ZHMS-A: vr. beat 6.764 min, m/z 415.1 [M+H] ⁺ Method AA 10 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2,5-bis(2,2,2-trifluoroethoxy)benzenesulfonamide ZHMS-A: vr. beat 6.785 min, m/z 505.0 [M+H] ⁺ Method AA 11 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-(oxazol-5-yl)benzenesulfonamide ZHMS-A: vr. beat 6.322 min, m/z 376.1 [M+H] ⁺ Method AA 12 methyl 2-( N- (4-chlorobenzo[ d ]isoxazol-3-yl)sulfamoyl)benzoate ZHMS-A: vr. beat 6.484 min, m/z 367.1 [M+H] ⁺ Method AA 13 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-3-methyl-2-oxo-2,3-dihydrobenzo[ d ]oxazole-6-sulfonamide ZHMS-A: vr. beat 6.103 min, m/z 378.0 [MH] ^- Method AA 14 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-3,4-dihydro-2 H -benzo[ b ][1,4]dioxepin-7-sulfonamide ZHMS-A: vr. beat 6.362 min, m/z 381.1 [M+H] ⁺ Method AA 15 N- (4-chlorobenzo[ d ]isoxazol-3-yl)benzo[ d ]thiazole-4-sulfonamide ZHMS-A: vr. beat 6.212 min, m/z 366.0 [M+H] ⁺ Method AA 16 N- (4-chlorobenzo[ d ]isoxazol-3-yl)benzo[ c ][1,2,5]oxadiazole-4-sulfonamide ZHMS-A: vr. beat 6.973 min, m/z 351.0 [M+H] ⁺ Method AA 17 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-(4-fluorophenoxy)benzenesulfonamide ZHMS-A: vr. beat 6.781 min, m/z 419.1 [M+H] ⁺ Method AA 18 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-methyl-5-(2-methyloxazol-5-yl)benzenesulfonamide ZHMS-A: vr. beat 6.410 min, m/z 404.1 [M+H] ⁺ Method AA 19 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-5-(isoxazol-5-yl)-2-methylbenzenesulfonamide ZHMS-A: vr. beat 6.542 min, m/z 390.1 [M+H] ⁺ Method AA 20 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-methyl-5-( 1H- pyrazol-1-yl)benzenesulfonamide ZHMS-A: vr. beat 6.493 min, m/z 389.1 [M+H] ⁺ Method AA 21 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-(methylsulfonyl)benzenesulfonamide ZHMS-A: vr. beat 6.317 min, m/z 385.0 [MH] ^- Method AA 22 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-isopropylbenzene sulfonamide ZHMS-A: vr. beat 6.745 min, m/z 351.1 [M+H] ⁺ Method AA 23 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-(trifluoromethoxy)benzenesulfonamide ZHMS-A: vr. beat 6.763 min, m/z 391.0 [MH] ^- Method AA 24 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-isopropoxybenzenesulfonamide ZHMS-A: vr. beat 6.620 min, m/z 367.1 [M+H] ⁺ Method AA 25 4-( tert -butyl)- N- (4-chlorobenzo[ d ]isoxazol-3-yl)benzenesulfonamide ZHMS-A: vr. beat 6.826 min, m/z 365.1 [M+H] ⁺ Method AA 26 4-chloro- N- (4-chlorobenzo[ d ]isoxazol-3-yl)benzenesulfonamide ZHMS-A: vr. beat 6.614 min, m/z 343.0, 345.0 [M+H] ⁺ Method AA 27 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-methylbenzenesulfonamide ZHMS-A: vr. beat 6.447 min, m/z 321.0 [MH] ^- AC Method 28 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-( 1H- pyrazol-1-yl)benzenesulfonamide ZHMS-A: vr. beat 6.321 min, m/z 375.1 [M+H] ⁺ AC Method 29 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2,6-difluorobenzene sulfonamide ZHMS-A: vr. beat 6.657 min, m/z 343.0 [MH] ^- AC Method 30 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-fluorobenzenesulfonamide ZHMS-A: vr. beat 6.389 min, m/z 327.0 [M+H] ⁺ AC Method 31 3-chloro- N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-methylbenzenesulfonamide ZHMS-A: vr. beat 6.808 min, m/z 355.0, 357.0 [MH] ^- AC Method 32 4-bromo- N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-(trifluoromethoxy)benzenesulfonamide ZHMS-A: vr. beat 7.440 min, m/z 468.9, 470.9 [MH] ^- AC Method 33 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-methoxy-4,5-dimethylbenzenesulfonamide ZHMS-A: vr. beat 6.538 min, m/z 367.1 [M+H] ⁺ AC Method 34 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-(difluoromethoxy)benzenesulfonamide ZHMS-A: vr. beat 6.487 min, m/z 373.0 [MH] ^- AC Method 35 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-4-(pentafluoro-λ ⁶ -sulfanyl)benzene sulfonamide ZHMS-A: vr. beat 7.021 min; m/z 435.0 [M+H] ⁺ AD Method 36 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-3-(pentafluoro-λ ⁶ -sulfanyl)benzene sulfonamide ZHMS-A: vr. beat 7.219 min; m/z 435.0 [M+H] ⁺ AD Method 37 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-5-isopropyl-2-methoxy-4-methylbenzenesulfonamide ZHMS-A: vr. beat 7.537 min; m/z 395.1 [M+H] ⁺ AD Method 38 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-(difluoromethoxy)benzenesulfonamide ZHMS-A: vr. beat 6.695 min; m/z 375.0 [M+H] ⁺ AD Method 39 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2,3-dihydrobenzo[ b ][1,4]dioxin-5-sulfonamide ZHMS-A: vr. beat 6,620 min; m/z 367.0 [M+H] ⁺ AD Method 40 5-( tert -butyl)- N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide ZHMS-B: vr. beat 3.876 min, m/z 395.2 [M+H] ⁺ AD Method 41 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-5-isopropyl-2-methoxybenzenesulfonamide ZHMS-A: vr. beat 6.684 min, m/z 381.1 [M+H] ⁺ AD Method 42 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-5,6,7,8-tetrahydronaphthalene-2-sulfonamide ZHMS-B: vr. beat 3.859 min, m/z 363.1 [M+H] ⁺ AD Method 43 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-[1,1'-biphenyl]-3-sulfonamide ZHMS-B: vr. beat 3.823 min, m/z 385.1 [M+H] ⁺ AD Method 44 N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2,3-dihydro-1 H -indene-5-sulfonamide ZHMS-B: vr. beat 3.785 min, m/z 349.1 [M+H] ⁺ AE Method 45 5-bromo- N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide ZHMS-B: vr. beat 3.752 min, m/z 417.0, 419.0 [M+H] ⁺ AF Method

Examples 46-71 (Table D):

BA Method

A mixture of benzo[ d ]isoxazol-3-amine and sulfonyl chloride in pyridine (1 mL) was stirred at room temperature for 16 h. The reaction mixture was concentrated, diluted with 5% aqueous HCl (1 mL), and sonicated for a minimum of 30 min. The resulting precipitate was collected by filtration or extraction with DCM (2 × 1 mL) and purified by silica gel column chromatography (EtOAc/petroleum gasoline gradient 40-60 °C) or preparative HPLC with mass separation to afford the desired product. For reaction components and amounts used, as well as purification conditions, see Table B.

BB Method

A mixture of benzo[ d ]isoxazol-3-amine and sulfonyl chloride in pyridine (0.5 mL) was stirred at room temperature for 64 h. The reaction mixture was concentrated, diluted with 5% aqueous HCl (1 mL), and sonicated for a minimum of 30 min. The resulting precipitate was collected by filtration, and a portion of the crude material (50 mg or less) was purified by preparative HPLC with mass separation to afford the desired product. For reaction components and amounts used, see Table B.

Method BC

A mixture of benzo[ d ]isoxazol-3-amine and sulfonyl chloride in pyridine (0.5 mL) was stirred at room temperature for 16 h. The reaction mixture was concentrated, diluted with 5% aqueous HCl (1 mL), and sonicated for a minimum of 30 min. The resulting precipitate was collected by filtration, and a portion of the crude material (50 mg or less) was purified by preparative HPLC with mass separation to afford the desired product. For reaction components and amounts used, see Table B.

Table B

Product Benzenesulfonyl chloride Benz[ d ] isoxazol-3-amine Method/processing/cleaning method Structure and name Weight (g) Moles (mmol) Weight (g) Moles (mmol) 46 0.143 0.602 0,030 0.23 Method BA/filtration/column chromatography, gradient 0-45% 47 0,150 0.682 0.031 0.23 Method BA/filtration/column chromatography, gradient 0-40% 48 0.148 0.563 0.033 0.25 Method BA/filtration/column chromatography, gradient 0-40% 49 0,140 0.561 0.031 0.23 BA Method/Filtration/
column chromatography, gradient 0-40% 50 0.142 0.535 0,030 0.22 BA Method/Filtration/
column chromatography, gradient 0-100% 51 0.156 0.611 0.033 0.24 BA Method/Filtration/
column chromatography, gradient 0-100% 52 0.036 0,180 0.028 0.21 Method BA/DCM extraction/column chromatography, gradient 0-50% 53 0.074 0.360 0.032 0.24 Method BA/DCM extraction/column chromatography, gradient 0-50% 54 0.148 0.54 0.034 0.25 BA Method/Filtration/
column chromatography, gradient 0-100% 55 0.158 0.58 0,030 0.22 BA Method/Filtration/
column chromatography, gradient 0-100% 56 0.063 0.27 0,030 0.22 Method BA/DCM extraction/column chromatography, gradient 0-100% 57 0,170 0.68 0.027 0.20 BA Method/Filtration/
column chromatography, gradient 0-40%. Triturated minimal amount of acetone, the product contained approx. 10% by-product sulfonic acid 58 0.101 0.488 0.029 0.21 BA/filtration/HPLC method with mass separation 59 0.105 0.466 0.033 0.25 BB Method 60 0.171 0.698 0.032 0.24 BB Method 61 0,100 0,640 0.032 0.24 BB Method 62 0,170 0.694 0.031 0.23 BB Method 63 0.107 0.454 0.033 0.25 Method BC 64 0,120 0.553 0.029 0.22 Method BC 65 0.103 0.447 0.032 0.24 Method BC 66 0,111 0,490 0,030 0.23 Method BC 67 0.1025 0.396 0.033 0.25 Method BC 68 0,111 0.432 0.033 0.25 BB Method

CA Method

A mixture of benzo[ d ]isoxazol-3-amine and sulfonyl chloride in pyridine (1 mL) was stirred at room temperature for 16 h, after which a second portion of benzenesulfonyl chloride was added and stirred for an additional 64 h. The reaction mixture was concentrated, diluted with 5% aqueous HCl (1 mL), and sonicated for a minimum of 30 min. The resulting precipitate was collected by filtration and purified either by preparative mass-separated HPLC (up to 50 mg crude material) or by silica gel column chromatography (0-40% EtOAc/petroleum gasoline, 40-60 °C) to afford the desired product. For reaction components and amounts used, as well as purification conditions, see Table C.

Table C

Product Benzenesulfonyl chloride (added in 2 parts) Benz[ d ] isoxazol-3-amine Method/method of purification Structure and name Part Weight (g) Moles (mmol) Weight (g) Moles (mmol) 69 1st 0.117 0.463 0,060 0.45 CA/HPLC method with mass separation 2nd 0.103 0.408 70 1st 0.092 0.36 0.052 0.39 CA method/column chromatography 2nd 0.088 0.35 71 1st 0.103 0.398 0.065 0.48 CA method/column chromatography 2nd 0.101 0.390

Table D

Structure Name Analytical data 46 N- (benzo[ d ]isoxazol-3-yl)-2,5-dimethoxybenzenesulfonamide LCMS-B: sp temp 3.560 min; m/z 335.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 10.01 (br s, 1H), 8.14-8.06 (m, 1H), 7.64 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.55 (dt, J= 8.5, 0.8 Hz, 1H), 7.43-7.36 (m, 2H), 7.17 (dd, J= 9.1, 2.9 Hz, 1H), 7.13 (d, J= 8.8 Hz, 1H), 3.82 (s, 3H), 3.77 (s, 3H) 47 N- (benzo[ d ]isoxazol-3-yl)-2-methoxy-5-methylbenzenesulfonamide LCMS-B: sp temp 3.585 min; m/z 319.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone -d ₆ ) δ 9.94 (br s, 1H), 8.14-8.06 (m, 1H), 7.70-7.65 (m, 1H), 7.63 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.54 (dt, J= 8.5, 0.8 Hz, 1H), 7.43-7.36 (m, 2H), 7.08 (d, J= 8.5 Hz, 1H), 3.84 (s, 3H), 2.28 (s, 3H). 48 N- (benzo[ d ]isoxazol-3-yl)-5-( tert -butyl)-2-methoxybenzenesulfonamide LCMS-B: sp temp 3.806 min; m/z 361.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 10.00 (br s, 1H), 8.13-8.08 (m, 1H), 7.91 (d, J= 2.5 Hz, 1H), 7.66-7.60 (m, 2H), 7.55-7.51 (m, 1H), 7.38 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H), 7.10 (d, J= 8.7 Hz, 1H), 3.83 (s, 3H), 1.26 (s, 9H) 49 N- (benzo[ d ]isoxazol-3-yl)-5-isopropyl-2-methoxybenzenesulfonamide ZHMS-B: vr. beat 3.744 min; m/z 347.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 9.97 (br s, 1H), 8.12-8.08 (m, 1H), 7.75 (d, J= 2.4 Hz, 1H), 7.63 (ddd, J= 8.4, 7.0, 1.2 Hz, 1H), 7.53 (dt, J= 8.5, 0.9 Hz, 1H), 7.48 (ddd, J= 8.5, 2.4, 0.6 Hz, 1H), 7.38 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H), 7.10 (d, J= 8.6 Hz, 1H), 3.84 (s, 3H), 2.95-2.87 (m, 1H), 1.18 (d, J= 6.9 Hz, 6H) 50 methyl 3-( N- (benzo[ d ]isoxazol-3-yl)sulfamoyl)-4-methoxy benzoate ZHMS-A: vr. beat 6.024 min; m/z 363.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 10.24 (br s, 1H), 8.55 (d, J = 2.2 Hz, 1H), 8.24 (dd, J = 8.8, 2.2 Hz, 1H), 8.09 (dt, J = 8.1, 1.0 Hz, 1H), 7.66 (ddd, J = 8.4, 7.0, 1.2 Hz, 1H), 7.57 (dt, J = 8.5, 0.8 Hz, 1H), 7.42 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H), 7.35 (d, J = 8.8 Hz, 1H), 4.00 (s, 3H), 3.89 (s, 3H) 51 N- (benzo[ d ]isoxazol-3-yl)-5-chloro-2-methoxy-4-methylbenzenesulfonamide LCMS-B: sp temp 3.738 min; m/z 353.1/355.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.10-8.06 (m, 1H), 7.81 (s, 1H), 7.65 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.58-7.54 (m, 1H), 7.43-7.37 (m, 1H), 7.21 (s, 1H), 3.87 (s, 3H), 2.38 (s, 3H) 52 N- (benzo[ d ]isoxazol-3-yl)-3-ethylbenzene sulfonamide LCMS-B: sp temp 3.707 min; m/z 303.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.02-7.97 (m, 1H), 7.85-7.82 (m, 1H), 7.82-7.78 (m, 1H), 7.68-7.62 (m, 1H), 7.59-7.55 (m, 1H), 7.54-7.45 (m, 2H), 7.41-7.35 (m, 1H), 2.70 (q, J= 7.6 Hz, 2H)*, 1.19 (t, J= 7.6 Hz, 3H). *Partially overlaps with the aqueous peak 53 N- (benzo[ d ]isoxazol-3-yl)-4-ethylbenzenesulfonamide LCMS-B: sp temp 3.690 min; m/z 303.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.03-7.98 (m, 1H), 7.93-7.89 (m, 2H), 7.65 (ddd, J= 8.2, 7.0, 1.1 Hz, 1H), 7.60-7.55 (m, 1H), 7.46-7.41 (m, 2H), 7.41-7.36 (m, 1H), 2.71 (q, J= 7.6 Hz, 2H)*, 1.21 (t, J= 7.6 Hz, 3H). *Partially overlaps with the aqueous peak 54 N- (benzo[ d ]isoxazol-3-yl)-2-methoxy-5-(1 H -pyrazol-1-yl)benzene sulfonamide ZHMS-B: vr. beat 3.554 min; m/z 371.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.36 (d, J= 2.8 Hz, 1H), 8.33-8.31 (m, 1H), 8.13-8.07 (m, 1H), 8.05 (dd, J= 9.0, 2.8 Hz, 1H), 7.68-7.61 (m, 2H), 7.56-7.52 (m, 1H), 7.40 (ddd, J= 8.0, 7.1, 0.8 Hz, 1H), 7.33 (d, J= 9.0 Hz, 1H), 6.49 (dd, J= 2.5, 1.7 Hz, 1H), 3.93 (s, 3H). 55 N- (benzo[ d ]isoxazol-3-yl)-5-(isoxazol-5-yl)-2-methoxybenzenesulfonamide LCMS-B: sp 3.516 min; m/z 372.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.46 (d, J = 1.9 Hz, 1H), 8.36 (d, J = 2.3 Hz, 1H), 8.13-8.07 (m, 2H), 7.63 (ddd, J = 8.3, 7.1, 1.2 Hz, 1H), 7.56-7.52 (m, 1H), 7.43-7.36 (m, 2H), 6.90 (d, J = 1.9 Hz, 1H), 3.97 (s, 3H) 56 N- (benzo[ d ]isoxazol-3-yl)-2,5-diethylbenzenesulfonamide ZHMS-B: vr. beat 3,801 min; m/z 331.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.01-7.97 (m, 1H), 7.95-7.92 (m, 1H), 7.66-7.61 (m, 1H), 7.57-7.52 (m, 1H), 7.45-7.41 (m, 1H), 7.40-7.34 (m, 2H), 3.10 (q, J= 7.5 Hz, 2H)*, 2.66 (q, J= 7.5 Hz, 2H)*, 1.22 (t, J= 7.5 Hz, 3H), 1.17 (t, J= 7.6 Hz, 3H). *Partially overlaps with the water peak 57 N- (benzo[ d ]isoxazol-3-yl)-2-ethoxy-5-ethylbenzenesulfonamide ZHMS-B: vr. beat 3.763 min; m/z 347.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 9.78 (br s, 1H), 8.13-8.08 (m, 1H), 7.74 (d, J= 2.3 Hz, 1H), 7.63 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.56-7.52 (m, 1H), 7.44-7.41 (m, 1H), 7.38 (ddd, J= 8.0, 7.1, 0.8 Hz, 1H), 7.09 (d, J= 8.5 Hz, 1H), 4.16 (q, J= 7.0 Hz, 2H), 1.29 (t, J= 7.0 Hz, 3H), 1.16 (t, J= 7.6 Hz, 3H). NB: about 10% sulfonic acid impurities 58 N- (benzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide HPLC-MS: sp 5.11 min; m/z 305.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 10.35-9.70 (br s, 1H), 8.12-8.08 (m, 1H), 7.90-7.86 (dd, J= 7.9, 1.7 Hz, 1H), 7.66-7.58 (m, 2H), 7.56-7.52 (m, 1H), 7.41-7.36 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H), 7.21-7.17 (dd, J= 8.4, 0.8 Hz, 1H), 7.09-7.03 (m, 1H), 3.90-3.87 (s, 3H). 59 N- (benzo[ d ]isoxazol-3-yl)-3-chloro-2-methylbenzenesulfonamide HPLC-MS: sp temp 6.38 min; m/z 323.22/325.17 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 11.10-10.01 (br s, 1H), 8.15-8.10 (dd, J= 8.0, 1.0 Hz, 1H), 8.02-7.97 (m, 1H), 7.74-7.69 (m, 1H), 7.68-7.62 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.59-7.54 (m, 1H), 7.46-7.41 (m, 1H), 7.41-7.37 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H), 2.77-2.74 (s, 3H) 60 N- (benzo[ d ]isoxazol-3-yl)-2,3-dichlorobenzenesulfonamide HPLC-MS: sp 6.27 min; m/z 343.13/345.15/347.10 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.25-8.22 (dd, J= 8.0, 1.5 Hz, 1H), 8.06-8.02 (m, 1H), 7.93-7.90 (dd, J= 8.1, 1.5 Hz, 1H), 7.69-7.64 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.64-7.59 (m, 1H), 7.59-7.55 (m, 1H), 7.44-7.38 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H) 61 N- (benzo[ d ]isoxazol-3-yl)-3,4-dichlorobenzenesulfonamide HPLC-MS: sp temp 6.62 min; m/z 343.13/345.15/347.10 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.19-8.17 (d, J = 2.1 Hz, 1H), 7.99-7.94 (m, 2H), 7.85-7.82 (d, J = 8.5 Hz, 1H), 7.70-7.65 (ddd, J = 8.2, 7.0, 1.2 Hz, 1H), 7.62-7.58 (m, 1H), 7.43-7.37 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H) 62 N- (benzo[ d ]isoxazol-3-yl)-2,4-dichlorobenzenesulfonamide HPLC-MS: sp temp 6.42 min; m/z 343.13/345.15/347.16 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.26-8.22 (d, J = 8.6 Hz, 1H), 8.06-8.02 (m, 1H), 7.75-7.72 (d, J = 2.0 Hz, 1H), 7.70-7.63 (m, 2H), 7.60-7.56 (m, 1H), 7.44-7.38 (ddd, J = 8.0, 7.0, 0.9 Hz, 1H) 63 N- (benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide HPLC-MS: sp 5.39 min; m/z 333.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 10.19-9.73 (br s, 1H), 8.13-8.08 (ddd, J= 8.1, 1.0, 1.0 Hz, 1H), 7.73-7.69 (d, J= 2.3 Hz, 1H), 7.66-7.60 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.56-7.52 (ddd, J= 8.5, 0.8, 0.8 Hz, 1H), 7.47-7.42 (dd, J= 8.5, 2.3 Hz, 1H), 7.42-7.36 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H), 7.13-7.07 (d, J= 8.5 Hz, 1H), 3.86-3.82 (s, 3H), 2.65-2.56 (k, J= 7.6 Hz, 2H), 1.18-1.12 (t, J= 7.6 Hz, 3H) 64 N- (benzo[ d ]isoxazol-3-yl)-2,3-dihydro-1 H -indene-5-sulfonamide HPLC-MS: sp 5.45 min; m/z 315.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 10.80-9.43 (br s, 1H), 8.04-7.99 (ddd, J= 8.1, 1.0, 1.0 Hz, 1H), 7.84-7.80 (m, 1H), 7.78-7.73 (m, 1H), 7.68-7.62 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.60-7.54 (ddd, J= 8.5, 0.8, 0.8 Hz, 1H), 7.42-7.35 (m, 2H), 2.97-2.88 (t, J= 7.5 Hz, 4H), 2.12-2.05 (dd, J= 15.0, 7.6 Hz, 2H). 65 N- (benzo[ d ]isoxazol-3-yl)-5,6,7,8-tetrahydronaphthalene-2-sulfonamide HPLC-MS: sp 5.57 min; m/z 329.5 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 10.44-9.90 (br s, 1H), 8.03-7.98 (ddd, J= 8.1, 1.1, 1.1 Hz, 1H), 7.69-7.63 (m, 3H), 7.60-7.55 (ddd, J= 8.5, 0.8, 0.8 Hz, 1H), 7.42-7.36 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H), 7.25-7.20 (d, J= 8.3 Hz, 1H), 2.81-2.76 (m, 4H), 1.83-1.73 (m, 4H). 66 N- (benzo[ d ]isoxazol-3-yl)naphthalene-2-sulfonamide HPLC-MS: sp 5.46 min; m/z 325.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 11.03-9.58 (br s, 1H), 8.66-8.62 (m, 1H), 8.15-8.11 (m, 1H), 8.11-8.08 (d, J= 8.9 Hz, 1H), 8.04-7.98 (m, 3H), 7.73-7.60 (m, 3H), 7.56-7.52 (ddd, J= 8.5, 0.8, 0.8 Hz, 1H), 7.41-7.35 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H). 67 N- (benzo[ d ]isoxazol-3-yl)-3-(5-methyl-1,3,4-oxadiazol-2-yl)benzenesulfonamide HPLC-MS: sp 5.00 min; m/z 357.7 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.62-8.61 (dd, J= 1.6, 1.6 Hz, 1H), 8.29-8.26 (ddd, J= 7.8, 1.4, 1.4 Hz, 1H), 8.23-8.19 (ddd, J= 8.0, 1.8, 1.1 Hz, 1H), 7.99-7.96 (ddd, J= 8.1, 0.9, 0.9 Hz, 1H), 7.85-7.80 (dd, J= 7.9, 7.9 Hz, 1H), 7.67-7.63 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.58-7.55 (m, 1H), 7.41-7.36 (ddd, J= 7.9, 7.0, 0.8 Hz, 1H), 2.63-2.60 (s, 3H). 68 N- (benzo[ d ]isoxazol-3-yl)-5-bromopyridin-2-sulfonamide ZHMS-B: vr. beat 3.583 min; m/z 354/356 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 11.19-10.23 (br s, 1H), 8.81-8.78 (dd, J= 2.3, 0.7 Hz, 1H), 8.38-8.34 (dd, J= 8.4, 2.3 Hz, 1H), 8.15-8.10 (dd, J= 8.4, 0.7 Hz, 1H), 8.06-8.01 (ddd, J= 8.1, 1.0, 1.0 Hz, 1H), 7.69-7.64 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.59-7.56 (ddd, J= 8.5, 0.8, 0.8 Hz, 1H), 7.43-7.38 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H). 69 N- (benzo[ d ]isoxazol-3-yl)-[1,1'-biphenyl]-4-sulfonamide HPLC-MS: time beat 6.62 min; m/z 351.19 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone _-d6 ) δ 8.11-8.06 (m, 2H), 8.04-8.00 (m, 1H), 7.90-7.85 (m, 2H), 7.76-7.68 (m, 2H), 7.66 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.58 (dt, J= 8.5, 0.8 Hz, 1H), 7.53-7.47 (m, 2H), 7.46-7.37 (m, 2H). 70 N- (benzo[ d ]isoxazol-3-yl)-[1,1'-biphenyl]-3-sulfonamide LCMS-B: sp temp 3.754 min; m/z 351.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.29-8.26 (m, 1H), 8.03-8.00 (m, 1H), 7.97 (dddd, J= 9.0, 7.8, 1.8, 0.9 Hz, 2H), 7.72-7.63 (m, 4H), 7.58 (dt, J= 8.5, 0.8 Hz, 1H), 7.54-7.48 (m, 2H), 7.46-7.37 (m, 2H) 71 N- (benzo[ d ]isoxazol-3-yl)-4-cyclohexyl benzenesulfonamide ZHMS-B: vr. beat 3,999 min; m/z 357.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.02-7.98 (m, 1H), 7.93-7.89 (m, 2H), 7.65 (ddd, J= 8.3, 7.0, 1.2 Hz, 1H), 7.59-7.55 (m, 1H), 7.47-7.43 (m, 2H), 7.38 (ddd, J= 8.0, 7.0, 0.9 Hz, 1H), 2.66-2.58 (m, 2H), 1.85-1.78 (m, 4H), 1.76-1.69 (m, 1H), 1.50-1.34 (m, 4H), 1.32-1.24 (m, 1H).

Examples 72-88 (Table E):

EA Method

To a solution of 4-methoxybenzo[ d ]isoxazol-3-amine (50 mg, 0.305 mmol) in THF (3.0 mL) was added NaH (60% in mineral oil, 49 mg, 1.22 mmol) and stirred at room temperature for 10 min. Sulfonyl chloride (1 eq, 0.305 mmol) was added and the reaction was stirred for 16 h. The volume of volatiles was reduced to approximately 1 mL, after which DCM (3 mL) and water (3 mL) were added and the mixture was stirred for 10 min. The mixture was passed through a phase separator, then the organic fraction was loaded onto a 1 g amine-modified silica gel cartridge (Biotage), the cartridge was washed with MeOH (6 mL), and the product was eluted with HCl (1.25 M in methanol, 6 mL). The HCl washings were evaporated in vacuo to obtain the desired product.

EB Method

To a solution of 4-methoxybenzo[ d ]isoxazol-3-amine (50 mg, 0.305 mmol) in DMF (3.0 mL) was added NaH (60% in mineral oil, 61 mg, 1.52 mmol) and stirred at room temperature for 10 min. Sulfonyl chloride (1 eq, 0.305 mmol) was added and the reaction was stirred for 16 h. The resulting mixture was loaded onto silica gel and purified by column chromatography (0-100% petroleum gasoline 40-60 °C, then 0-60% MeOH in EtOAc) to give the desired product.

EC Method

To a solution of 4-methoxybenzo[ d ]isoxazol-3-amine (50 mg, 0.305 mmol) in DMF (5 mL) was added NaH (60% in mineral oil, 22 mg, 0.914 mmol) and stirred at room temperature for 10 min. Sulfonyl chloride (1 eq, 0.305 mmol) was added and the reaction was stirred for 16 h. The resulting mixture was quenched with water (3 mL), stirred for 10 min at room temperature, then loaded onto silica gel and purified by column chromatography (0-100% petroleum gasoline, 40-60 °C, then 0-60% MeOH in EtOAc) to give the desired product.

ED Method

To a solution of 4-methoxybenzo[ d ]isoxazol-3-amine (100 mg, 0.609 mmol) in THF (5.0 mL) was added NaH (60% in mineral oil, 5 or 10 equiv) and stirred at room temperature for 10 min. Sulfonyl chloride (1 equiv, 0.609 mmol) was added and the reaction was stirred for 16 h. The resulting mixture was loaded onto silica gel and purified by column chromatography (0-100% petroleum gasoline 40-60 °C, then 0-60% MeOH in EtOAc) to give the desired product.

EF Method

To a solution of 4-methoxybenzo[ d ]isoxazol-3-amine (100 mg, 0.609 mmol) in THF (5.0 mL) was added NaH (60% in mineral oil, 122 mg, 3.05 mmol) and stirred at room temperature for 10 min. Sulfonyl chloride (1 eq, 0.609 mmol) was added and the reaction was stirred for 16 h. The resulting mixture was loaded onto silica gel and purified by column chromatography (0-100% petroleum gasoline 40-60 °C, then 0-60% MeOH in EtOAc), the isolated solids were sonicated in MeOH (1 mL) and collected by filtration to give the desired product.

EG Method

To a solution of 4-methoxybenzo[ d ]isoxazol-3-amine (100 mg, 0.609 mmol) in THF (5.0 mL) was added NaH (60% in mineral oil, 122 mg, 3.05 mmol) and stirred at room temperature for 10 min. Sulfonyl chloride (1 eq, 0.609 mmol) was added and the reaction was stirred for 16 h at room temperature. The volume of volatiles was reduced to approximately 1 mL, after which DCM (3 mL) and water (3 mL) were carefully added and stirred for 10 min. The mixture was passed through a phase separator, then the organic fraction was loaded onto a 1 g amine-modified silica gel cartridge (Biotage), the cartridge was washed with MeOH (6 mL), and the product was eluted with HCl (2 M, 1:1 methanol:1,4-dioxane, 6 mL). The HCl washes were then evaporated in vacuo to give the desired product.

EH Method

A suspension of 4-methoxybenzo[ d ]isoxazol-3-amine (48 mg, 0.29 mmol) and NaH (60% in mineral oil, 0.117 mg, 2.93 mmol) in DMF (10 mL) was stirred at room temperature for 10 min and then cooled to -78 °C. To this cooled mixture was added sulfonyl chloride (1.5 equiv, 0.439 mmol) and the mixture was stirred at -78 °C for 1 h, then warmed to room temperature and stirred for 16 h. The reaction mixture was loaded onto silica gel and purified by column chromatography (Biotage Isolera, ₂₄ g SiO2 cartridge, 0-100% EtOAc in petroleum gasoline 40-60 °C, then 0-40% MeOH in EtOAc) to give a solid that was suspended in diethyl ether (25 mL) and sonicated for 5 min. The solid was collected by filtration and air dried to give the desired product.

EI Method

A mixture of 4-methoxybenzo[ d ]isoxazol-3-amine (0.035 g, 0.21 mmol) and sulfonyl chloride (1.05 equiv, 0.22 mmol) in pyridine (1 mL) was stirred at room temperature for 16 h. The reaction mixture was concentrated, diluted with 5% aqueous HCl (1 mL), and sonicated for a minimum of 30 min. Extraction with DCM (2 x 1 mL) and purification by silica gel column chromatography (0-100% EtOAc in petroleum gasoline, 40-60 °C) provided the desired product.

EJ Method

To a solution of 4-methoxybenzo[ d ]isoxazol-3-amine (1 eq.) in THF (3 mL) was added LiHMDS (1 M in THF, 1.5 eq.). After 10 min of stirring, sulfonyl chloride (1.5 eq.) was added and the reaction mixture was allowed to stir for 17 h in open air. THF was removed in vacuo, followed by the addition of DCM (3 mL) and water (3 mL) and stirring for 10 min. After separation of the layers, the organic fraction was loaded onto a 1 g amine-modified silica gel cartridge (Biotage). The cartridge was washed with MeOH (6 mL) and then stripped with 1.25 M HCl in MeOH (6 mL). The HCl washings were then evaporated in vacuo to afford the desired product.

Table E

Structure and output (if applicable) Name Analytical data Method 72
15 mg, 13% 2-methoxy- N- (4-methoxybenzo[ d ]isoxazol-3-yl)-4,6-dimethylbenzene sulfonamide ZHMS-A: vr. beat 6.474 min, m/z 363.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.51-9.42 (s, 1H), 7.61-7.51 (t, J= 8.2 Hz, 1H), 7.19-7.10 (d, J= 8.4 Hz, 1H), 6.90-6.87 (s, 1H), 6.87-6.82 (d, J= 8.0 Hz, 1H), 6.80-6.74 (s, 1H), 3.99-3.93 (s, 3H), 3.84-3.78 (s, 3H), 2.58-2.54 (s, 3H), 2.31-2.25 (s, 3H). EA Method 73
10 mg, 8% N- (4-methoxy benzo[ d ]isoxazol-3-yl)-3-methyl quinoline-8-sulfonamide ZHMS-A: vr. beat 6.469 min, m/z =370.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ=8.94 (d, J= 2.1 Hz, 1H), 8.37 (dd, J= 7.3, 1.3 Hz, 1H), 8.33 (s, 1H), 8.25 (d, J= 8.0 Hz, 1H), 7.76 (t, J= 7.8 Hz, 1H), 7.50 (t, J= 8.2 Hz, 1H), 7.10 (d, J= 8.4 Hz, 1H), 6.79 (d, J= 8.1 Hz, 1H), 3.89 (s, 3H), 2.52 (s, 3H). EB Method 74
35 mg, 29% 5-methoxy- N- (4-methoxybenzo[ d ] isoxazol-3-yl)quinoline-8-sulfonamide ZHMS-A: vr. beat 6.461 min, m/z 386.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ=9.08 (dd, J= 4.3, 1.7 Hz, 1H), 8.62 (dd, J= 8.5, 1.7 Hz, 1H), 8.42 (d, J= 8.4 Hz, 1H), 7.67 (dd, J= 8.5 Hz, 4.3, 1H), 7.49 (t, J= 8.2 Hz, 1H), 7.22 (d, J= 8.5 Hz, 1H), 7.08 (d, J= 8.4 Hz, 1H), 6.78 (d, J= 8.0 Hz, 1H), 4.08 (s, 3H), 3.94 (s, 3H) EC Method 75
29 mg, 24% 4-methoxy- N- (4-methoxybenzo[ d ]isoxazol-3-yl)quinoline-8-sulfonamide ZHMS-A: vr. beat 6.253 min, m/z 386.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ=8.93 (d, J= 5.4 Hz, 1H), 8.52-8.37 (m, 2H), 7.75 (t, J= 7.8 Hz, 1H), 7.50 (t, J= 8.3 Hz, 1H), 7.22 (d, J= 5.3 Hz, 1H), 7.10 (d, J= 8.6 Hz, 1H), 6.80 (d, J= 8.1 Hz, 1H), 4.08 (s, 3H), 3.93 (s, 3H) EC Method 76
59 mg, 26% 2,4-dimethoxy- N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide ZHMS-A: vr. beat 6.188 min, m/z =365.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ=9.76 (s, 1H), 7.80-7.73 (m, 1H), 7.55 (t, J= 8.2 Hz, 1H), 7.15 (d, J= 8.4 Hz, 1H), 6.84 (d, J= 8.0 Hz, 1H), 6.73-6.62 (m, 2H), 3.92 (s, 3H), 3.84 (s, 3H), 3.79 (s, 3H) ED Method 10 equiv NaH were used 77
68 mg, 30% N- (4-methoxy benzo[ d ]isoxazol-3-yl)-7-methylquinoline-8-sulfonamide ZHMS-A: vr. beat 6.419 min, m/z =370.2 [M+H] ⁺ EF Method 78
4 mg, 2% N- (4-methoxy benzo[ d ]isoxazol-3-yl)-6-methylquinoline-8-sulfonamide ZHMS-A: vr. beat 6.332 min, m/z =370.1 [M+H] ⁺ ED Method 5 equiv NaH were used 79
15 mg, 7% N- (4-methoxy benzo[ d ]isoxazol-3-yl)-4-methylquinoline-8-sulfonamide ZHMS-A: vr. beat 6.325 min, m/z =370.1 [M+H] ⁺ ED Method 5 equiv NaH were used 80
50 mg, 22% N- (4-methoxy benzo[ d ]isoxazol-3-yl)-2-methylquinoline-8-sulfonamide ZHMS-A: vr. beat 6.349 min, m/z =370.1 [M+H] ⁺ ED Method 5 equiv NaH were used 81
38 mg, 18% N- (4-methoxy benzo[ d ]isoxazol-3-yl)benzofuran - 7-sulfonamide ZHMS-A: vr. beat 6.225 min, m/z =345.1 [M+H] ⁺ EG Method 82
44 mg, 20% N- (4-methoxy benzo[ d ]isoxazol-3-yl)chroman-8-sulfonamide ZHMS-A: vr. beat 6.282 min, m/z =361.1 [M+H] ⁺ EG Method 83
10 mg, 9% N- (4-methoxy benzo[ d ]isoxazol-3-yl)quinoline-8-sulfonamide ZHMS-A: vr. beat 6,198 min; m/z =356.2 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ=9.08 (dd, J= 4.3, 1.7 Hz, 1H), 8.57 (dd, J= 8.4, 1.6 Hz, 1H), 8.46 (dd, J= 7.4, 1.4 Hz, 1H), 8.36 (d, J= 8.0 Hz, 1H), 7.81 (t, J= 7.8 Hz, 1H), 7.73 (dd, J= 8.3, 4.3 Hz, 1H), 7.50 (t, J= 8.2 Hz, 1H), 7.10 (d, J= 8.4 Hz, 1H), 6.78 (d, J= 8.0 Hz, 1H), 3.88 (s, 3H) EH Method 84
9 mg, 12% N- (4-methoxy benzo[ d ]isoxazol-3-yl)-5,6,7,8-tetrahydronaphthalene-2-sulfonamide LCMS-A: sp temp 6.955 min; m/z 359.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, acetone- d ₆ ) δ 7.81-7.76 (m, 2H), 7.55-7.49 (m, 1H), 7.23 (d, J= 7.7 Hz, 1H), 7.06 (d, J= 8.4 Hz, 1H), 6.79 (d, J= 8.0 Hz, 1H), 3.96 (s, 3H), 1.81-1.77 (m, 4H). NB: solvent obscures aliphatic 2xCH ₂ EI Method 85 2,4,6-trimethoxy- N- (4-methoxy benzo[ d ]isoxazol-3-yl)benzene sulfonamide ZHMS-A: vr. beat 6.095 min; m/z 395.1 [M+H] ⁺ EJ Method 86 2,6-dimethoxy- N- (4-methoxy benzo[ d ]isoxazol-3-yl)benzenesulfonamide ZHMS-A: vr. beat 5,991 min; m/z 365.1 [M+H] ⁺ EJ Method 87 2-methoxy- N- (4-methoxybenzo[ d ]isoxazol-3-yl)-5-(trifluoromethyl)benzenesulfonamide ZHMS-B: vr. beat 3.379 min; m/z 403.0 [M+H] ⁺ EJ Method 88 N- (4-methoxy benzo[ d ]isoxazol-3-yl)-2-(methoxy methyl)benzenesulfonamide ZHMS-B: vr. beat 3,344 min; m/z 349.1 [M+H] ⁺ EJ Method

Examples 89-147 (Table F)

EJ Method

To a solution of the amine (0.5 mmol, 1.0 equiv.) in anhydrous THF (10 mL) at -78 °C under N ₂ was added dropwise LiHMDS (1 M solution in THF, 3 equiv.) and the mixture was stirred at -78 °C for 30 min. Then a solution of sulfonyl chloride (1.5 equiv.) in anhydrous THF (2.0 mL) was added dropwise and the mixture was allowed to warm to rt and stirred overnight. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography or prep. TLC to give the title compound. Variations of the above conditions are noted in Table F.

FB Method

To a solution of the amine (0.3 mmol, 1.0 equiv) in pyridine (5 mL) under _N2 was added sulfonyl chloride (2.0 equiv) and the mixture was heated at 100 °C overnight. The reaction was quenched with 1 M aq. HCl, followed by the addition of water and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography or prep. TLC to give the title compound. Variations of the above conditions are noted in Table F.

FC Method

To a solution of the amine (0.5 mmol, 1.0 equiv.) in anhydrous THF (10 mL) at -20 °C under N ₂ was added n -BuLi (2.5 M in hexanes, 1.5 equiv.) dropwise and the mixture was stirred at -20 °C for 1 h. Then a solution of sulfonyl chloride (1.5 equiv.) in anhydrous THF (2.0 mL) was added dropwise and the mixture was allowed to warm to rt and stirred overnight. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH = 20/1) to give the title compound.

FD Method

To a solution of the amine (0.5 mmol, 1.0 equiv.) in anhydrous THF (10 mL) at -78 °C under N ₂ was added dropwise KHMDS (1 M solution in THF, equivalents listed in Table F) and the mixture was stirred at -78 °C or 0 °C for a period of 30 min to 1 h (listed in Table F). Then a solution of the sulfonyl chloride (equivalents listed in Table F) in anhydrous THF (2.0 mL) was added dropwise and the mixture was allowed to warm to rt and stirred overnight. Water was added and the mixture was extracted with EtOAc. The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography or prep. TLC to give the title compound.

Table F

Name and structure Analytical data Intermediate connections (if applicable) Method Notes 89
N -(6-Bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide JHMS-C: vr. _Ud. 2.18 min, m/z 442.9 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.80 (s, 1H), 7.54-7.48 (m, 2H), 7.05 (s, 1H), 6.78 (d, J= 8.4 Hz, 2H), 3.92 (s, 3H), 3.76 (s, 6H). 2,6-Dimethoxybenzenesulfonyl chloride I111
6-Bromo-4-methoxybenzo[ d ]isoxazol-3-amine I22 FA 4 equiv. LiHMDS were used.
Rev. TLC (DCM/MeOH=100/1) 90
N -(4-Methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-C: _sp 1.97 min, m/z 304.9 [M+H] ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.21 (d, J = 7.2 Hz, 2H), 7.82 (s, 1H), 7.61-7.54 (m, 3H), 7.43 (t, J = 8.0 Hz, 1H), 7.02 (d, J = 8.0 Hz, 1H), 6.59 (d, J = 7.6 Hz, 1H), 3.91 (s, 3H). FA The mixture was stirred for 2 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=100/1) 91
3,5-Dimethoxy -N- (4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)-[1,1'-biphenyl]-4-sulfonamide JHMS-C: vr. _beat 2.33 min, m/z 485.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.26 (s, 1H), 7.52 (d, J= 6.8 Hz, 2H), 7.46-7.42 (m, 3H), 6.98 (s, 1H), 6.73 (s, 2H), 6.65 (s, 1H), 4.51 (s, 2H), 4.04 (s, 3H), 3.94 (s, 6H), 3.41 (s, 3H). 3,5-Dimethoxy-[1,1'-biphenyl]-4-sulfonyl chloride I116
4-Methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I9 FA 5 equiv. LiHMDS were used and stirred for 2 h before adding the sulfonyl chloride.
Column chromatography (pet. ether/EtOAc=2/1) 92
N -(6-(3-Hydroxyphenyl)-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide JHMS-C: vr. _beat 2.19 min, m/z 457.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.64 (s, 1H), 9.62 (s, 1H), 7.53 (t, J= 8.8 Hz, 1H), 7.35 (s, 1H), 7.30 (t, J= 8.0 Hz, 1H), 7.19 (d, J= 7.6 Hz, 1H), 7.12 (s, 1H), 7.01 (s, 1H), 6.85-6.78 (m, 3H), 4.01 (s, 3H), 3.79 (s, 6H). 2,6-Dimethoxybenzenesulfonyl chloride I111
3-(3-Amino-4-methoxybenzo[ d ]isoxazol-6-yl)phenol I19 FA 8 equiv. LiHMDS were used and stirred for 1 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=30/1) 93
2,6-Dimethoxy -N- (4-methoxy-6-(1-methoxyethyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide JHMS-C: vr. _beat 1.94 min, m/z 423.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.56 (s, 1H), 7.51 (t, J= 8.4 Hz, 1H), 7.08 (s, 1H), 6.80 (d, J= 8.4 Hz, 2H), 6.78 (s, 1H), 4.45 (k, J= 6.4 Hz, 1H), 3.92 (s, 3H), 3.78 (s, 6H), 3.17 (s, 3H), 1.37 (d, J= 6.4 Hz, 3H). 2,6-Dimethoxybenzenesulfonyl chloride I111
4-Methoxy-6-(1-methoxyethyl)benzo[ d ]isoxazol-3-amine I15 FA The mixture was stirred for 2 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=75/1) 94
3-Ethyl-2,6-dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-C: _sp 2.31 min, m/z 393.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 9.57 (s, 1H), 7.57 (t, J= 8.0 Hz, 1H), 7.46 (d, J= 8.8 Hz, 1H), 7.16 (d, J= 8.8 Hz, 1H), 6.93 (d, J= 8.8 Hz, 1H), 6.85 (d, J= 8.0 Hz, 1H), 3.92 (s, 3H), 3.78 (s, 3H), 3.75 (s, 3H), 2.60 (k, J= 7.2 Hz, 2H), 1.15 (t, J= 7.2 Hz, 3H). 3-Ethyl-2,6-dimethoxybenzenesulfonyl chloride I106 FA The mixture was stirred for 3 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=75/1) 95
3,5-Dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)-[1,1'-biphenyl]-4-sulfonamide LCMS-C: _sp 2.34 min, m/z 441.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.60 (s, 1H), 7.79 (d, J= 6.8 Hz, 2H), 7.58-7.44 (m, 4H), 7.17 (d, J= 8.0 Hz, 1H), 6.99 (s, 2H), 6.87 (d, J= 7.6 Hz, 1H), 3.95 (s, 3H), 3.88 (s, 6H). 3,5-Dimethoxy-[1,1'-biphenyl]-4-sulfonyl chloride I116 FA 3.3 equiv of LiHMDS were used and stirred at -70°C for 3 h before adding 1.8 equiv of sulfonyl chloride.
Purification: precipitated from DCM using pet. ether 96
2,4-Dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)-[1,1'-biphenyl]-3-sulfonamide JHMS-C: vr. _beat 2.42 min, m/z 441.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.84 (s, 1H), 7.59-7.54 (m, 2H), 7.48-7.42 (m, 4H), 7.39-7.35 (m, 1H), 7.18 (d, J= 8.0 Hz, 1H), 7.07 (d, J= 8.8 Hz, 1H), 6.87 (d, J= 8.0 Hz, 1H), 3.91 (s, 3H), 3.81 (s, 3H), 3.34 (s, 3H). 2,4-Dimethoxy-[1,1'-biphenyl]-3-sulfonyl chloride I114 FA 2 equiv. LiHMDS were used and
stirred for 3 hours before adding sulfonyl chloride.
Rev. TLC (DCM/MeOH=50/1) 97
2,6-Dimethoxy -N- (6-(methoxymethyl)-5-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: vr. _beat 2.38 min, m/z 393.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.3 (s, 1H), 7.81 (s, 1H), 7.48-7.45 (m, 2H), 6.75-6.73 (m, 2H), 4.51 (s, 2H), 3.73 (s, 6H), 3.35 (s, 3H), 2.31 (s, 3H). 2,6-Dimethoxybenzenesulfonyl chloride I111
6-(Methoxy
methyl)-5-methylbenzo[ d ]isoxazol-3-amine I4 FA 2 equiv. LiHMDS were used and stirred for 2.5 h before adding 3 equiv. sulfonyl chloride.
Purified using prep. HPLC 98
7-Methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)quinoline-8-sulfonamide LCMS-D: _sp 1.79 min, m/z 385.8 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.6 (s, 1H), 8.98 (dd, J= 4.4, 1.6 Hz, 1H), 8.46 (dd, J= 8.0, 1.6 Hz, 1H), 8.31 (d, J= 5.6 Hz, 1H), 7.55 (dd, J= 8.0, 4.0 Hz, 1H), 7.52 (d, J= 6.0 Hz, 1H), 7.50 (t, J= 8.0 Hz, 1H), 7.08 (d, J= 8.4 Hz, 1H), 6.81 (d, J= 8.0 Hz, 1H), 4.06 (s, 3H), 3.99 (s, 3H). 7-Methoxyquinoline-8-sulfonyl chloride I107 FA At -60°C, LiHMDS was added and stirred at 0°C for 1 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=20/1) 99
2-Methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)-5-phenoxybenzenesulfonamide LCMS-D: _sp 2.91 min, m/z 427.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.79 (d, J= 8.8 Hz, 2H), 7.56 (t, J= 8.4 Hz, 1H), 7.15-6.99 (m, 7H), 6.82 (d, J= 8.0 Hz, 1H), 3.90 (s, 3H), 3.76 (s, 3H). 2-Methoxy-5-phenoxybenzenesulfonyl chloride I100 FA 1.1 equiv. LiHMDS and 1.05 equiv. sulfonyl chloride were used.
The product has not been cleaned. 100
2,6-Dimethoxy -N- (5-(trifluoromethoxy)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: _sp 2.61 min, m/z 419.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, methanol- d ₄ ) δ 8.01 (d, J= 0.8 Hz, 1H), 7.59-7.42 (m, 3H), 6.71 (d, J= 8.8 Hz, 2H), 3.82 (s, 6H). 2,6-Dimethoxybenzenesulfonyl chloride I111
5-(Trifluoromethoxy)benzo[ d ]isoxazol-3-amine I63 FA At -60°C, LiHMDS was added and stirred at 0°C for 1 h before adding the sulfonyl chloride.
Prep. TLC (pet. ether/EtOAc=5/1) 101
2,6-Dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)-3-(trifluoromethyl)benzenesulfonamide LCMS-D: vr. _beat . 2.64 min, m/z 433.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 10.5 (s, 1H), 7.92 (d, J= 8.8 Hz, 1H), 7.59 (t, J= 8.0 Hz, 1H), 7.20-7.14 (m, 2H), 6.85 (d, J= 8.4 Hz, 1H), 3.87 (s, 6H), 3.85 (s, 3H). 2,6-Dimethoxy-3-(trifluoromethyl)benzenesulfonyl chloride I102 FA 2 equiv LiHMDS were added at 0 °C.
Prep. TLC (pet. ether/EtOAc=1/1) 102
2,6-Dimethoxy -N- (5-methyl-6-(oxazol-2-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: vr. _beat 2.42 min, m/z 415.9 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 11.5 (s, 1H), 8.32 (s, 1H), 7.07 (t, J= 8.4 Hz, 2H), 7.50-7.30 (m, 2H), 6.76 (d, J= 8.4 Hz, 2H), 3.75 (s, 6H), 2.70 (s, 3H). 2,6-Dimethoxybenzenesulfonyl chloride I111
5-Methyl-6-(oxazol-2-yl)benzo[ d ]isoxazol-3-amine I65 FA 1.5 equiv. LiHMDS were used.
Prep. TLC (pet. ether/EtOAc=1/2) 103
2,6-Dimethoxy -N- (6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: vr. _beat 2.22 min, m/z 378.9 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.4 (s, 1H), 8.04 (d, J= 8.4 Hz, 1H), 7.50-7.45 (m, 2H), 7.31 (d, J= 8.4 Hz, 1H), 6.75 (d, J= 8.8 Hz, 2H), 4.55 (s, 2H), 3.73 (s, 6H), 3.32 (s, 3H). 2,6-Dimethoxybenzenesulfonyl chloride I111
6-(Methoxymethyl)benzo[d]isoxazol-3-amine I62 FA At -78°C, LiHMDS was added and stirred at 0°C for 30 min before adding the sulfonyl chloride.
Prep. TLC (pet. ether/EtOAc=2/1) 104
2,6-Dimethoxy -N- (5-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: vr. _beat 2.40 min, m/z 348.9 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.3 (s, 1H), 7.83 (s, 1H), 7.49-7.44 (m, 3H), 6.75 (d, J= 8.4 Hz, 2H), 3.74 (s, 6H), 2.40 (s, 3H). 2,6-Dimethoxybenzenesulfonyl chloride I111
5-Methylbenzo[ d ]isoxazol-3-amine I60 FA At -60°C, LiHMDS was added and stirred at 0°C for 1 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=20/1) 105
N -(6-(3,5-Dimethyl-1 H -pyrazol-1-yl)benzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide LCMS-D: vr. _beat 2.44 min, m/z 429.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.5 (s, 1H), 8.16 (d, J= 8.8 Hz, 1H), 7.74 (d, J= 1.6 Hz, 1H), 7.57 (dd, J= 8.8, 2.0 Hz, 1H), 7.48 (t, J= 8.8 Hz, 1H), 6.75 (d, J= 8.4 Hz, 2H), 6.13 (s, 1H), 3.75 (s, 6H), 2.36 (s, 3H), 2.19 (s, 3H). 2,6-Dimethoxybenzenesulfonyl chloride I111
6-(3,5-Dimethyl- 1H -pyrazol-1-yl)benzo[ d ]isoxazol-3-amine I59 FC 106
4,6-Dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)-2,3-dihydro-1 H -indene-5-sulfonamide LCMS-D: vr. _beat 2.58 min, m/z 405.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 9.40 (s, 1H), 7.58 (t, J= 8.0 Hz, 1H), 7.16 (d, J= 8.0 Hz, 1H), 6.86-6.84 (m, 2H), 3.96 (s, 3H), 3.80 (s, 3H), 3.75 (s, 3H), 2.88-2.84 (m, 4H), 2.02-1.99 (m, 2H). 4,6-Dimethoxy-2,3-dihydro- 1H -indene-5-sulfonyl chloride I98 FA 1.33 equiv LiHMDS was added at -78°C and stirred at 0°C for 1 h before adding 0.67 equiv sulfonyl chloride.
Column chromatography (pet. ether/EtOAc=1/1) 107
N -(4-Chlorobenzo[ d ]isoxazol-3-yl)-5-ethyl-1-methyl-2-oxo-1,2-dihydropyridine-3-sulfonamide LCMS-D: vr. _beat 2.34 min, m/z 368.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 10.5 (s, 1H), 7.98 (s, 2H), 7.70-7.62 (m, 2H), 7.45 (d, J= 7.2 Hz, 1H), 3.50 (s, 3H), 2.46 (k, J= 7.2 Hz, 2H), 1.13 (t, J= 7.2 Hz, 3H). 5-Ethyl-1-methyl-2-oxo-1,2-dihydropyridine-3-sulfonyl chloride I94 FA 2.1 equiv LiHMDS were added at -78°C and stirred at 0°C for 1 h before adding 2 equiv sulfonyl chloride.
Column chromatography (DCM/MeOH=50/1) 108
N -(5-Bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: vr. _beat 2.82 min, m/z 441.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 10.8 (s, 1H), 7.86 (d, J= 8.8 Hz, 1H), 7.62 (d, J= 1.2 Hz, 1H), 7.50-7.43 (m, 2H), 7.17 (d, J= 8.8 Hz, 1H), 3.91 (s, 3H), 3.79 (s, 3H), 2.64 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
5-Bromo-4-methoxybenzo[ d ]isoxazol-3-amine I48 FD 2 equiv KHMDS were added and stirred at 0°C for 1 h before adding 2 equiv sulfonyl chloride.
Column chromatography (pet. ether/EtOAc=50/1 to 3/1) 109
N -(5-Ethoxybenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: _sp 2.78 min, m/z 377.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.5 (s, 1H), 7.69 (d, J= 2.0 Hz, 1H), 7.50-7.44 (m, 3H), 7.20 (dd, J= 9.8, 2.4 Hz, 1H), 7.10 (d, J= 8.8 Hz, 1H), 4.06 (k, J= 7.4 Hz, 2H), 3.74 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.38 (t, J= 6.8 Hz, 3H), 1.16 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
5-Ethoxy benzo[ d ]isoxazol-3-amine I57 FB Column chromatography (pet. ether/EtOAc=4/1 to 3/1) 110
5-Ethyl-2-methoxy -N- (4-(methoxymethyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: _sp 2.66 min, m/z 377.1 [M+H] ⁺ ; 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
4-(Methoxymethyl)benzo[ d ]isoxazol-3-amine I51 FB Prep. TLC (pet. ether/EtOAc=2/1) 111
5-Ethyl-2-methoxy -N- (5-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: vr. _beat 2.65 min, m/z 363.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.5 (s, 1H), 7.69 (d, J= 2.0 Hz, 1H), 7.52-7.44 (m, 3H), 7.23 (dd, J= 9.8, 2.4 Hz, 1H), 7.10 (d, J= 8.8 Hz, 1H), 3.79 (s, 3H), 3.75 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
5-Methoxybenzo[ d ]isoxazol-3-amine I54 FB Prep. TLC 112
N -(4-Ethoxybenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide JHMS-E: vr. _beat 5.57 min m/z 376.7 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.68 (s, 1H), 7.67 (d, J= 2.0 Hz, 1H), 7.56-7.45 (m, 2H), 7.16-7.08 (m, 2H), 6.83 (d, J= 8.0 Hz, 1H), 4.19 (k, J= 7.2 Hz, 2H), 3.63 (s, 3H), 2.64 (k, J= 7.6 Hz, 2H), 1.31 (t, J= 7.4 Hz, 3H), 1.17 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
4-Ethoxy benzo[ d ]isoxazol-3-amine I53 FA 1.5 equiv LiHMDS was added at -78°C and stirred at 0°C for 1 h before adding 1.5 equiv sulfonyl chloride.
Column chromatography (pet. ether/EtOAc=3/1) 113
N -(5-Bromobenzo[ d ]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonamide LCMS-D: _sp 3.07 min, m/z 437.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 11.6 (s, 1H), 8.25 (s, 1H), 7.78 (dd, J= 8.8, 2.0 Hz, 1H), 7.62-7.56 (m, 2H), 6.85 (s, 1H), 3.69 (s, 3H), 2.73-2.69 (m, 4H), 1.71 (m, 4H). 3-Methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonyl chloride I109
5-Bromo
benzo[ d ]isoxazol-3-amine I40 FA 2 equiv LiHMDS were added at -78°C and stirred at 0°C for 1 h before adding 2 equiv sulfonyl chloride.
Column chromatography (DCM/MeOH=200/1 to 100/1) 114
5-Bromo -N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-methylbenzenesulfonamide LCMS-D: _sp 2.82 min, m/z 401.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.02 (d, J= 2.0 Hz, 1H), 7.73-7.76 (m, 3H), 7.43-7.35 (m, 2H), 2.49 (s, 3H). FA 1 equiv LiHMDS was added at -78°C and stirred at 0°C for 1 h before adding 0.67 equiv sulfonyl chloride.
Column chromatography (pet. ether/EtOAc=6/1) 115
Methyl 4-chloro-2-( N -(4-chlorobenzo[ d ]isoxazol-3-yl)sulfamoyl)benzoate LCMS-D: _sp 2.78 min, m/z 401.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.09 (d, J= 2.0 Hz, 1H), 7.88 (d, J= 8.4 Hz, 1H), 7.74-7.65 (m, 3H), 7.48 (d, J= 7.6 Hz, 1H), 3.64 (s, 3H). FD 0.67 equiv KHMDS was added and stirred at 0°C for 1 h before adding 0.67 equiv sulfonyl chloride.
Column chromatography (pet. ether/EtOAc=1/1) 116
5-Ethyl-2-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: vr. _beat 2.56 min, m/z 363.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.96 (s, 1H), 7.66 (d, J= 1.6 Hz, 1H), 7.57 (t, J= 8.0 Hz, 1H), 7.48-7.46 (m, 1H), 7.12-7.11 (m, 2H), 6.84 (d, J= 8.0 Hz, 1H), 3.87 (s, 3H), 3.75 (s, 3H), 2.64 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112 FA 2 equiv LiHMDS were added at -78°C and stirred at 0°C for 1 h before adding 2 equiv sulfonyl chloride.
Column chromatography (DCM/MeOH=30/1) 117
N -(4-Chlorobenzo[ d ]isoxazol-3-yl)-2,3,4-trifluorobenzenesulfonamide LCMS-D: _sp 2.59 min, m/z 363.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.74-7.69 (m, 1H), 7.54-7.40 (m, 3H), 7.31 (d, J= 7.0 Hz, 1H). FA 1 equiv LiHMDS was added at -78°C and stirred at 0°C for 1 h before adding 1 equiv sulfonyl chloride.
Column chromatography (DCM/MeOH=100/1 to 30/1) 118
N -(4-Chlorobenzo[ d ]isoxazol-3-yl)-3-methylquinoline-8-sulfonamide LCMS-D: _sp 2.62 min, m/z 374.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.81 (d, J= 2.4 Hz, 1H), 8.34-8.25 (m, 3H), 7.76-7.61 (m, 3H), 7.43 (d, J= 8.8 Hz, 1H), 2.50 (s, 3H). FA 1.5 equiv LiHMDS was added at -78°C and stirred at 0°C for 1 h before adding 1.5 equiv sulfonyl chloride.
Column chromatography (DCM/MeOH=20/1) 119
N -(4-Chlorobenzo[ d ]isoxazol-3-yl)-6-methoxy-2,3-dihydro-1 H -indene-5-sulfonamide LCMS-D: vr. _beat 2.69 min, m/z 379.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.4 (s, 1H), 7.75-7.63 (m, 2H), 7.58 (s, 1H), 7.47 (d, J= 7.2 Hz, 1H), 7.11 (s, 1H), 3.69 (s, 3H), 2.93 (t, J= 7.2 Hz, 2H), 2.84 (t, J= 7.2 Hz, 2H), 2.07-2.00 (m, 2H). 6-Methoxy-2,3-dihydro- 1H -indene-5-sulfonyl chloride I108 FD 0.67 equiv KHMDS was added and stirred at 0°C for 1 h before adding 0.67 equiv sulfonyl chloride.
Column chromatography (pet. ether/EtOAc=1/1) 120
N -(4-Chlorobenzo[ d ]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonamide LCMS-D: vr. _beat 2.86 min, m/z 393.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.4 (s, 1H), 7.73-7.64 (m, 2H), 7.48 (d, J= 7.6 Hz, 1H), 7.45 (s, 1H), 6.91 (s, 1H), 3.67 (s, 3H), 2.78 (m, 2H), 2.67 (m, 2H), 1.72 (m, 4H). 3-Methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonyl chloride I109 FA 1 equiv LiHMDS was added at -78°C and stirred at 0°C for 1 h before adding 1 equiv sulfonyl chloride.
Column chromatography (pet. ether/EtOAc=50/1 to 2/1) 121
5-Ethyl-2-methoxy -N- (4-(trifluoromethyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: vr. _beat 2.74 min, m/z 401.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 10.3 (s, 1H), 8.14 (t, J= 6.4 Hz, 1H), 7.91-7.88 (m, 2H), 7.55 (d, J= 2.4 Hz, 1H), 7.51 (dd, J= 8.8, 2.4 Hz, 1H), 7.20 (d, J= 8.4 Hz, 1H), 3.82 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
4-(Trifluoromethyl)benzo[ d ]isoxazol-3-amine I42 FA 2 equiv LiHMDS were added at -78°C and stirred at 0°C for 1 h before adding 2 equiv sulfonyl chloride.
Column chromatography (DCM/MeOH=20/1) 122
N -(4-Chlorobenzo[ d ]isoxazol-3-yl)quinoxaline-5-sulfonamide LCMS-D: _sp 2.30 min, m/z 361.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 11.0 (s, 1H), 9.09 (d, J= 1.4 Hz, 1H), 9.08 (d, J= 1.4 Hz, 1H), 8.47-8.43 (m, 2H), 8.05 (t, J= 8.0 Hz, 1H), 7.72-7.62 (m, 2H), 7.42 (d, J= 7.8 Hz, 1H). FA 2 equiv. LiHMDS were used.
Purification: dissolved in 2 M aq NaOH, washed with EtOAc, then acidified to pH 2 and extracted with DCM 123
N -(4-Chlorobenzo[ d ]isoxazol-3-yl)quinoline-8-sulfonamide LCMS-D: vr. _beat 2.41 min, m/z 360.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 8.93 (dd, J= 4.3, 1.8 Hz, 1H), 8.58 (dd, J= 8.4, 1.8 Hz, 1H), 8.40-8.35 (m, 2H), 7.79 (t, J= 7.8 Hz, 1H), 7.73-7.60 (m, 3H), 7.41 (d, J= 7.4 Hz, 1H). FA 2 equiv. LiHMDS were used.
Purification: dissolved in 2 M aq NaOH, washed with EtOAc, then acidified to pH 2 and extracted with DCM 124
N -(4-Chlorobenzo[ d ]isoxazol-3-yl)naphthalene-1-sulfonamide LCMS-D: _sp 2.60 min, m/z 359.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.73 (t, J= 8.4 Hz, 1H), 8.31-8.24 (m, 2H), 8.14 (d, J= 7.8 Hz, 1H), 7.74-7.63 (m, 5H), 7.46 (d, J= 7.2 Hz, 1H). FA 2 equiv. LiHMDS were used.
Column chromatography (pet. ether/EtOAc=50/1 to 2/1 125
N -(6-(1 H -1,2,3-Triazol-1-yl)benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: vr. _beat 2.48 min, m/z 400.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.9 (s, 1H), 8.96 (s, 1H), 8.26-8.23 (m, 2H), 8.04-8.01 (m, 2H), 7.73 (d, J= 2.0 Hz, 1H), 7.49 (dd, J= 8.4, 2.0 Hz, 1H), 7.11 (d, J= 8.4 Hz, 1H), 3.73 (s, 3H), 2.65 (k, J= 8.0 Hz, 2H), 1.23 (t, J= 8.0 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
6-(1H - 1,2,3-Triazol-1-yl)benzo[ d ]isoxazol-3-amine I36 FB Column chromatography (DCM/MeOH=100/0 to 50/1) 126
5-Ethyl-2-methoxy -N- (6-(pyrimidin-2-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: _sp 2.65 min, m/z 411.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 8.98 (d, J= 4.8 Hz, 2H), 8.43 (d, J= 8.4 Hz, 1H), 8.48 (s, 1H), 8.20 (d, J= 8.4 Hz, 1H), 7.73 (d, J= 2.0 Hz, 1H), 7.55 (t, J= 4.8 Hz, 1H), 7.48 (dd, J= 8.4, 2.0 Hz, 1H), 7.11 (d, J= 8.8 Hz, 1H), 3.72 (s, 3H), 2.65 (k, J= 7.2 H, 2H), 1.17 (t, J= 7.2 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
6-(Pyrimidin-2-yl)benzo[ d ]isoxazol-3-amine I39 FB Rev. TLC (DCM/MeOH=50/1) 127
5-Bromo -N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2,3-dihydrobenzofuran-7-sulfonamide LCMS-D: vr. _beat 2.66 min,
m/z 428.9 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO -d ₆ ) δ 11.2 (s, 1H), 7.75-7.65 (m, 3H), 7.58 (d, J= 2.0 Hz, 1H), 7.48 (d, J= 7.2 Hz, 1H), 4.57 (t, J= 8.8 Hz, 2H), 3.26 (t, J= 8.8 Hz, 2H). 5-Bromo-2,3-dihydrobenzofuran-7-sulfonyl chloride I95 FA At -78°C, LiHMDS was added and stirred at 0°C for 1 h before adding the sulfonyl chloride.
Column chromatography (pet. ether/EtOAc=5/1) 128
N -(5-Bromo-4-chlorobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide JHMS-E: vr. _beat 5.81 min, m/z 444.7 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 10.7 (s, 1H), 8.02 (d, J= 8.8 Hz, 1H), 7.74 (d, J= 8.8 Hz, 1H), 7.60 (d, J= 2.4 Hz, 1H), 7.50-7.48 (m, 1H), 7.16 (d, J= 8.4 Hz, 1H), 3.66 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
5-Bromo-4-chlorobenzo[ d ]isoxazol-3-amine I44 FD 1 equiv KHMDS was added, stirred at -78°C for 30 min, then 2 equiv sulfonyl chloride was added.
Column chromatography (pet. ether/EtOAc=50/1 to 2/1) 129
N -(4-Bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: vr. _beat 2.69 min, m/z 410.9 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.4 (s, 1H), 7.78 (d, J= 8.0 Hz, 1H), 7.65-7.56 (m, 3H), 7.49 (dd, J= 8.4, 2.4 Hz, 1H), 7.15 (d, J= 8.4 Hz, 1H), 3.68 (s, 3H), 2.60 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
4-Bromobenzo[ d ]isoxazol-3-amine I41 FA 1.5 equiv. LiHMDS were used.
Column chromatography (pet. ether/EtOAc=2/1 to 1/1) 130
N -(6-(2 H -1,2,3-Triazol-2-yl)benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: vr. _beat 2.77 min, m/z 400.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.9 (s, 1H), 8.24-8.17 (m, 3H), 8.12-8.10 (m, 2H), 7.70 (s, 1H), 7.48 (d, J= 8.4 Hz, 1H), 7.11 (d, J= 8.4 Hz, 1H), 3.76 (s, 3H), 2.65 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
6-(2 H -1,2,3-Triazol-2-yl)benzo[ d ]isoxazol-3-amine I72 FB 3 equiv. of sulfonyl chloride were used.
Rev. TLC (DCM/MeOH=50/1) 131
N -(6-(1 H -Pyrazol-1-yl)benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: _sp 2.61 min, m/z 399.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO -d ₆ ) δ 11.8 (s, 1H), 8.66 (d, J= 2.4 Hz, 1H), 8.15 (d, J= 8.4 Hz, 1H), 8.07 (d, J= 2.0 Hz, 1H), 7.97 (dd, J= 8.8, 2.0 Hz, 1H), 7.83 (d, J= 1.2 Hz, 1H), 7.72 (d, J= 2.0 Hz, 1H), 7.48 (dd, J= 8.4, 2.0 Hz, 1H), 7.11 (d, J= 8.4 Hz, 1H), 6.62 (d, J= 1.2 Hz, 1H), 3.73 (s, 3H), 2.65 (k, J= 7.6 Hz, 2H), 1.18 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
6-(1 H -Pyrazol-1-yl)benzo[ d ]isoxazol-3-amine I70 FB 3 equiv. of sulfonyl chloride were used.
Column chromatography (DCM/MeOH=50/1) 132
N -(7-Ethoxybenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: _sp 2.67 min, m/z 377.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 7.69 (d, J= 2.0 Hz, 1H), 7.57 (d, J= 7.6 Hz, 1H), 7.46 (dd, J= 8.8, 2.0 Hz, 1H), 7.26 (d, J= 8.0 Hz, 1H), 7.15 (d, J= 8.0 Hz, 1H), 7.08 (d, J= 8.4 Hz, 1H), 4.21 (k, J= 6.8 Hz, 2H), 3.72 (s, 3H), 3.62 (k, J= 7.6 Hz, 2H), 1.36 (t, J= 6.8 Hz, 3H), 1.16 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
7-Ethoxybenzo[ d ]isoxazol-3-amine I26 FB Column chromatography (pet. ether/EtOAc=5/1) 133
N -(7-(Cyclopropylmethoxy)benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: _sp 2.87 min, m/z 403.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 7.69 (d, J= 1.6 Hz, 1H), 7.56 (d, J= 8.0 Hz, 1H), 7.46 (d, J= 8.0 Hz, 1H), 7.24 (t, J= 8.0 Hz, 1H), 7.14 (d, J= 7.6 Hz, 1H), 7.09 (d, J= 8.4 Hz, 1H), 3.99 (d, J= 7.2 Hz, 2H), 3.72 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.30 (m, 1H), 1.16 (t, J= 7.6 H, 3H), 0.59-0.54 (m, 2H), 0.35-0.30 (m, 2H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
7-(Cyclopropylmethoxy)benzo[ d ]isoxazol-3-amine I28 FB Column chromatography (DCM/MeOH=200/1) 134
N -(6-(Cyclopropylmethoxy) benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: _sp 2.89 min, m/z 403.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.5 (s, 1H), 7.87 (d, J= 8.8 Hz, 1H), 7.68 (d, J= 2.4 Hz, 1H), 7.46 (dd, J= 8.4, 2.0 Hz, 1H), 7.09-7.07 (m, 2H), 6.96 (dd, J= 8.8, 2.0 Hz, 1H), 3.88 (d, J= 6.8 Hz, 2H), 3.73 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.30 (m, 1H), 1.16 (t, J= 7.6 Hz, 3H), 0.60-0.55 (m, 2H), 0.25-0.22 (m, 2H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
6-(Cyclopropyl methoxy)benzo[ d ]isoxazol-3-amine I33 FB Purified using prep. HPLC 135
N -(5-Bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: vr. _beat 2.83 min, m/z 410.9 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 8.25 (s, 1H), 8.78 (d, J= 8.8 Hz, 1H), 7.71 (s, 1H), 7.62 (d, J= 8.8 Hz, 1H), 7.48 (d, J= 8.0 Hz, 1H), 7.11 (d, J= 8.4 Hz, 1H), 3.72 (s, 3H), 3.64 (k, J= 7.2 Hz, 2H), 1.17 (t, J= 7.2 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
5-Bromo
benzo[ d ]isoxazol-3-amine I40 FA 2 equiv. LiHMDS and 2 equiv. sulfonyl chloride were used.
Prep. TLC (pet. ether/EtOAc=2/1) 136
5-Ethyl-2-methoxy -N- (7-(pyrimidin-2-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: _sp 2.52 min, m/z 411.1 [M+H] ⁺ ; ^1H NMR (400 MHz, methanol _-d4 / _CDCl3 ) δ 8.87 (d, J= 4.8 Hz, 2H), 8.51 (d, J= 7.6 Hz, 1H), 8.15-8.13 (m, 1H), 7.76 (d, J= 2.4 Hz, 1H), 7.49 (t, J= 8.0 Hz, 1H), 7.37-7.33 (m, 2H), 6.95 (d, J= 8.8 Hz, 1H), 3.81 (s, 3H), 2.61 (k, J= 7.6 Hz, 2H), 1.18 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
7-(Pyrimidin-2-yl)benzo[ d ]isoxazol-3-amine I68 FB Prep. TLC (pet. ether/EtOAc=1/1) 137
N -(7-Bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: _sp 2.85 min, m/z 411.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.9 (s, 1H), 8.08 (d, J= 8.0 Hz, 1H), 7.89 (d, J= 7.2 Hz, 1H), 7.71 (d, J= 2.0 Hz, 1H), 7.48 (d, J= 8.4 Hz, 1H), 7.34 (d, J= 7.6 Hz, 1H), 7.10 (d, J= 8.4 Hz, 1H), 3.71 (s, 3H), 2.64 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
7-Bromobenzo[ d ]isoxazol-3-amine I66 FB 3 equiv. of sulfonyl chloride were used.
Prep. TLC (pet. ether/EtOAc=5/1) 138
N -(6-Bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: vr. _beat 2.93 min, m/z 411.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 8.08-7.99 (m, 2H), 7.70 (d, J= 2.0 Hz, 1H), 7.58 (dd, J= 8.8, 1.6 Hz, 1H), 7.48 (dd, J= 8.8, 2.0 Hz, 1H), 7.10 (d, J= 8.4 Hz, 1H), 3.71 (s, 3H), 2.64 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
6-Bromobenzo
[ d ]isoxazol-3-amine I75 FB 1.8 equiv of sulfonyl chloride were used.
Column chromatography (pet. ether/EtOAc=from 100/0 to 5/1) 139
N -(6-Ethoxybenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: _sp 2.76 min, m/z 377.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.5 (s, 1H), 7.88 (d, J= 8.8 Hz, 1H), 7.66 (d, J= 2.0 Hz, 1H), 7.47-7.44 (m, 1H), 7.11-7.08 (m, 1H), 6.94 (dd, J= 8.8, 2.0 Hz, 1H), 4.11 (k, J= 7.2 Hz, 2H), 3.73 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.36 (t, J= 7.2 Hz, 3H), 1.17 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
6-Ethoxy benzo[ d ]isoxazol-3-amine I32 FB Column chromatography (pet. ether/EtOAc=from 100/0 to 5/1) 140
5-Ethyl-2-methoxy -N- (6-(pyridin-2-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: _sp 2.63 min, m/z 410.1 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 8.74-8.69 (m, 1H), 8.26 (s, 1H), 8.18-8.09 (m, 3H), 7.98-7.90 (m, 1H), 7.72 (d, J= 2.3 Hz, 1H), 7.50-7.39 (m, 2H), 7.13-7.06 (m, 1H), 3.73 (s, 3H), 2.61 (k, J= 7.6 Hz, 2H), 1.15 (t, J= 7.6 Hz, 3H). 5-Ethyl-2-methoxybenzenesulfonyl chloride I112
6-(Pyridin-2-yl)benzo[ d ]isoxazol-3-amine I74 FB Column chromatography (DCM/MeOH=100/0 to 100/1) 141
N -(Benzo[ d ]isoxazol-3-yl)-4-bromo-2-methoxybenzenesulfonamide LCMS-D: _sp 2.58 min, m/z 382.9 [M+H] ⁺ ; 4-Bromo-2-methoxybenzenesulfonyl chloride I110 FB 1.2 equiv of sulfonyl chloride were used.
Column chromatography (pet. ether/EtOAc=5/1 to 2/1) 142
2,6-Dimethoxy- N -(4-methoxy-7-phenylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-C: _sp 2.39 min, m/z 441.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.62 (s, 1H), 7.90-7.69 (m, 3H), 7.59-7.32 (m, 4H), 6.97 (d, J = 8.4 Hz, 1H), 6.86-6.73 (m, 2H), 3.98 (s, 3H), 3.79 (s, 6H). 2,6-Dimethoxybenzenesulfonyl chloride I111
4-Methoxy-7-phenylbenzo[ d ]isoxazol-3-amine I80 FA 4 equiv. LiHMDS were used and stirred for 2 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=100/1) 143
2,6-Dimethoxy- N -(4-methoxy-7-(1-methyl-1 H -pyrazol-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide JHMS-C: vr. _beat 1.89 min, m/z 445.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 9.58 (s, 1H), 8.22 (s, 1H), 7.96 (s, 1H), 7.80 (d, J =8.2 Hz, 1H), 7.50 (t, J =8.5 Hz, 1H), 6.88 (d, J =8.3 Hz, 1H), 6.78 (d, J =8.5 Hz, 2H), 3.95 (s, 3H), 3.88 (s, 3H), 3.79 (s, 6H). 2,6-Dimethoxybenzenesulfonyl chloride I111
4-Methoxy-7-(1-methyl-1 H- pyrazol-4-yl)benzo[ d ]isoxazol-3-amine I82 FA 4 equiv. LiHMDS were used and stirred for 2 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=30/1) 144
2,6-Dimethoxy- N -(4-methoxy-6-(oxazol-2-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide JHMS-C: vr. _beat 1.81 min, m/z 431.9 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.81 (s, 1H), 8.33 (d, J =0.9 Hz, 1H), 7.73 (d, J =0.9 Hz, 1H), 7.51 (t, J =8.5 Hz, 1H), 7.48 (d, J =0.8 Hz, 1H), 7.38 (d, J =1.0 Hz, 1H), 6.79 (d, J =8.5 Hz, 2H), 4.01 (s, 3H), 3.78 (s, 6H). 2,6-Dimethoxybenzenesulfonyl chloride I111
4-Methoxy-6-(oxazol-2-yl)benzo[d]isoxazol-3-amine I86 FA The mixture was stirred for 2 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=75/1) 145
N -(5-Chloro-4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide JHMS-C: vr. _beat 2.27 min, m/z 442.9 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.4 (s, 1H), 7.56-7.49 (m, 2H), 6.80 (d, J =8.5 Hz, 2H), 4.58 (s, 2H), 3.93 (s, 3H), 3.78 (s, 6H), 3.41 (c, 3H). 2,6-Dimethoxybenzenesulfonyl chloride I111
5-Chloro-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine I83 FA 1.5 equiv. LiHMDS was used and stirred for 2 h before adding 2 equiv. sulfonyl chloride.
Rev. TLC (DCM/MeOH=100/1) 146
2,6-Dimethoxy- N -(4-methoxy-6-(pyridin-2-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide JHMS-C: vr. _beat 1.89 min, m/z 442.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.67 (s, 1H), 8.75-8.68 (m, 1H), 8.15 (d, J =8.0 Hz, 1H), 7.98-7.90 (m, 1H), 7.86 (s, 1H), 7.59 (s, 1H), 7.51 (t, J =8.5 Hz, 1H), 7.47-7.41 (m, 1H), 6.79 (d, J =8.6 Hz, 2H), 4.02 (s, 3H), 3.79 (s, 6H). 2,6-Dimethoxybenzenesulfonyl chloride I111
4-Methoxy-6-(pyridin-2-yl)benzo[ d ]isoxazol-3-amine I77 FA 4 equiv LiHMDS were used and stirred for 1 h before adding the sulfonyl chloride.
Rev. TLC (DCM/MeOH=50/1) 147
2,6-Dimethoxy- N -(6-(oxazol-2-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: vr. _beat 2.31 min, m/z 401.8 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 8.32 (s, 1H), 8.19-8.18 (m, 1H), 8.07 (s, 1H), 7.96 (d, J =8.4 Hz, 1H), 7.45-7.40 (m, 2H), 6.73 (d, J =8.4 Hz, 2H), 3.71 (s, 6H). 2,6-Dimethoxybenzenesulfonyl chloride I111
6-(Oxazol-2-yl)benzo[d]isoxazol-3-amine I90 FB Prep. TLC (pet. ether/EtOAc=1/1)

Example 148: N- (benzo[ d ]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide 148

A solution of 2,4-dimethoxybenzenesulfonyl chloride (0.18 g, 0.75 mmol) and benzo[ d ]isoxazol-3-amine (0.10 g, 0.75 mmol) in pyridine (1 mL) was irradiated in a microwave reactor at 110 °C for 2 h. The resulting mixture was loaded onto silica gel and the product was purified twice by column chromatography (4 g SiO ₂ cartridge, 0-45% EtOAc in petroleum gasoline 40-60 °C, then 4 g SiO ₂ cartridge, 0-35% EtOAc in petroleum gasoline 40-60 °C) to give two crops (78 mg and 5 mg) of the title compound (total weight 83 mg, yield 33%) as white solids. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.11 (d, J = 8.05 Hz, 1H), 7.79 (s, 1H), 7.70 (d, J = 8.81 Hz, 1H), 7.57-7.50 (m, 1H), 7.47-7.40 (m, 1H), 7.37-7.29 (m, 1H), 6.50 (d, J = 2.27 Hz, 1H), 6.42 (dd, J = 2.25, 8.81 Hz, 1H), 3.98 (s, 3H), 3.81 (s, 3H). LCMS-B: temp. spec. 3.20 min, m/z =356.8 [M+Na] ⁺ , 334.8 [M+H] ⁺ .

Example 149: N- (benzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 149

A solution of 2,6-dimethoxybenzene-1-sulfonyl chloride I111 (0.088 g, 0.37 mmol) and benzo[ d ]isoxazol-3-amine (0.050 g, 0.37 mmol) in pyridine (1 mL) was irradiated in a microwave reactor at 110 °C for 2 h and then at 120 °C for 2 h. The reaction mixture was loaded onto silica gel and purified by column chromatography (12 g SiO ₂ cartridge, 0-35% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (3.9 mg, 3.1% yield) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.30 (s, 1H), 8.17 (dt, J= 1.04, 8.15 Hz, 1H), 7.55-7.47 (m, 1H), 7.47-7.34 (m, 2H), 7.34-7.28 (m, 1H), 6.60 (d, J= 8.52 Hz, 2H), 3.91 (s, 6H). ZHMS-B: vr. beat 3.13 min, m/z =334.8 [M+H] ⁺ .

Example 150: N- (5-chlorobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 150

a) 5-Chlorobenzo[d]isoxazol-3-amine A1

To a suspension of acetohydroxamic acid (531 mg, 7.07 mmol) in DMF (10 mL) was added potassium tert -butoxide (793 mg, 7.07 mmol) and stirred at room temperature for 30 min. 5-Chloro-2-fluorobenzonitrile (1.00 g, 6.43 mmol) was added and the reaction mixture was heated to 50 °C for 1 h. After cooling, the reaction mixture was diluted with aqueous saturated NaCl solution (15 mL), the aqueous layer was extracted with EtOAc (3 × 100 mL), the organic layers were combined, dried (Na ₂ SO ₄ ), filtered and the volatiles were removed in vacuo. The residue was loaded onto silica gel and the product was purified by column chromatography (Biotage Isolera, 40 g SiO ₂ cartridge, 0-40% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound as a white solid (507 mg, 47%). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 7.94 (dd, J = 2.1, 0.6, 1H), 7.59-7.48 (m, 1H), 6.51 (s, 1H).

b) N-(5-chlorobenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 150

A suspension of 5-ethyl-2-methoxybenzene-1-sulfonyl chloride (150 mg, 0.639 mmol) and 5-chlorobenzo[ d ]isoxazol-3-amine A1 (108 mg, 0.639 mmol) in pyridine (1.5 mL) was irradiated in a microwave reactor at 110 °C for 2 h. 10 M aqueous KOH (1 mL) was added and the resulting mixture was stirred for 4 h at room temperature. The reaction mixture was loaded onto silica gel and the product was purified by column chromatography (0-100% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound as a white solid (53 mg, 23%). ¹ H NMR (400 MHz, DMSO _-d6 ) δ=8.11 (t, J= 1.4, 1H), 7.71 (d, J= 2.3, 1H), 7.68 (d, J= 1.4, 2H), 7.48 (dd, J= 8.5, 2.3, 1H), 7.10 (d, J= 8.6, 1H), 3.72 (s, 3H), 2.62 (k, J= 7.6, 2H), 1.16 (t, J= 7.6, 3H). ZHMS-A: vr. beat 6.637 min; m/z 367.0 [M+H] ⁺ .

Example 151: N- (4-chlorobenzo[ d ]isoxazol-3-yl)benzenesulfonamide 151

a) N-(4-chlorobenzo[d]isoxazol-3-yl)-N-(phenylsulfonyl)benzenesulfonamide A2

A solution of 4-chlorobenzo[ d ]isoxazol-3-amine (50 mg, 0.298 mmol) and benzenesulfonyl chloride (2 equiv, 0.595 mmol) in pyridine (1.5 mL) was irradiated in a microwave reactor for 2 h at 100 °C. After cooling, the reaction mixture was loaded onto silica gel and purified by silica gel column chromatography (Biotage Isolera, 24 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline 40-60 °C) to give the title compound. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ=7.97 (d, J= 8.6, 1H), 7.90-7.77 (m, 6H), 7.71-7.64 (m, 7H), 7.57 (d, J= 7.7, 1H).

b) N-(4-chlorobenzo[d]isoxazol-3-yl)benzenesulfonamide 151

A suspension of N- (4-chlorobenzo[ d ]isoxazol-3-yl) -N- (phenylsulfonyl)benzenesulfonamide A2 (50 mg, 0.11 mmol) in THF (10 mL) and 10 M aqueous KOH (1 mL) was stirred at room temperature for 16 h. The reaction mixture was diluted with water (25 mL), the aqueous layer was extracted with EtOAc (3×50 mL), the combined organic layers were washed with brine (25 mL) _, dried ( _Na2SO4 ), filtered and concentrated under reduced pressure. The resulting gum was dissolved in a minimum amount of acetone, petroleum naphtha 40-60 °C (50 mL) was added, the precipitate was filtered and air-dried to give the title compound as a tan solid (10 mg, 29%). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 7.89-7.80 (m, 2H), 7.41-7.30 (m, 4H), 7.26 (dd, J = 8.3, 0.8, 1H), 7.10 (dd, J = 7.5, 0.8, 1H). LCMS-A: sp 6.334 min, m/z 307.0 [MH] ^- .

Example 152: 5-ethyl- N- (7-fluorobenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 152

a) 7-fluorobenzo[d]isoxazol-3-amine A3

To a suspension of acetohydroxamic acid (594 mg, 7.91 mmol) in DMF (10 mL) was added potassium tert -butoxide (887 mg, 7.91 mmol) and the reaction mixture was stirred at room temperature for 30 min. 2,3-Difluorobenzonitrile (1.00 g, 7.19 mmol) was added and the reaction mixture was heated to 50 °C for 1 h. After cooling, the reaction mixture was diluted with aqueous saturated NaCl (15 mL), the aqueous layer was extracted with EtOAc (3 × 100 mL), the organic layers were combined, dried (Na ₂ SO ₄ ), filtered and the volatiles were removed in vacuo. The resulting gum was loaded onto silica gel and purified by column chromatography (Biotage Isolera, 24 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound as a white solid (303 mg, 27%). ¹ H NMR (400 MHz, CDCl ₃ ) δ = 7.33 (dd, J = 7.6, 1.3, 1H), 7.30-7.26 (m, 1H), 7.26-7.19 (m, 1H), 4.45 (s, 2H). LCMS-B: sp 3.371 min, m/z 153.2 [M+H] ⁺ .

b) 5-Ethyl-N-(7-fluorobenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide 152

A solution of 7-fluorobenzo[ d ]isoxazol-3-amine A3 (100 mg, 0.657 mmol) and 2-methoxy-5-ethylsulfonyl chloride I112 (154 mg, 0.657 mmol) in pyridine (2 mL) was irradiated in a microwave reactor for 2 h at 100 °C. After cooling, the reaction mixture was loaded onto silica gel and purified by column chromatography (Biotage Isolera, 24 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline 40-60 °C) to give the title compound as a white solid (127 mg, 55%). ¹ H NMR (400 MHz, DMSO _-d6 ) δ=7.91 (dd, J= 8.1, 0.8, 1H), 7.73 (d, J= 2.3, 1H), 7.57 (dd, J= 11.9, 8.0, 1H), 7.48 (dd, J= 8.5, 2.3, 1H), 7.38 (td, J= 8.0, 4.1, 1H), 7.10 (d, J= 8.6, 1H), 3.73 (s, 3H), 2.63 (k, J= 7.6, 2H), 1.16 (t, J= 7.6, 3H). ZHMS-A: vr. beat 6.429 min, m/z 351.1 [M+H] ⁺ .

Example 153: N- (4-chloro-5-methylbenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 153

a) 4-Chloro-5-methylbenzo[d]isoxazol-3-amine A4

To a suspension of acetohydroxamic acid (487 mg, 6.49 mmol) in DMF (10 mL) was added potassium tert -butoxide (728 mg, 6.49 mmol) and stirred at room temperature for 30 min. 2-Chloro-6-fluoro-3-methylbenzonitrile (1.00 g, 5.90 mmol) was added and the reaction mixture was heated to 50 °C for 1 h. After cooling, the reaction mixture was diluted with aqueous saturated NaCl solution (15 mL), the aqueous layer was extracted with EtOAc (3 × 100 mL), the organic layers were combined, dried (Na ₂ SO ₄ ), filtered and the volatiles were removed in vacuo. The resulting solid was sonicated in acetone (10 mL) followed by the addition of petroleum naphtha 40-60 °C (50 mL) and the precipitate collected by filtration and air dried to give the product as a white solid (524 mg, 49%). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 7.51 (d, J = 8.5, 1H), 7.38 (d, J = 8.5, 1H), 6.15 (s, 2H), 2.38 (s, 3H). LCMS-B: sp 3.562 min, m/z 183.1 [M+H] ⁺ .

b) N-(4-Chloro-5-methylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 153

A solution of 4-chloro-5-methylbenzo[ d ]isoxazol-3-amine A4 (100 mg, 0.548 mmol) and 2-methoxy-5-ethylsulfonyl chloride I112 (129 mg, 0.548 mmol) in pyridine (2 mL) was irradiated in a microwave reactor for 2 h at 100 °C. After cooling, the reaction mixture was added to water, the precipitate was removed by filtration, the filtrate was loaded onto silica gel and purified by column chromatography (Biotage Isolera, 24 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline 40-60 °C) to give the title compound as a white solid (33 mg, 16%). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ=10.48 (s, 1H), 7.69-7.57 (m, 3H), 7.48 (dd, J= 8.5, 2.3, 1H), 7.15 (d, J= 8.5, 1H), 3.68 (s, 3H), 2.60 (q, J= 7.5, 2H), 2.42 (s, 3H), 1.15 (t, J= 7.6, 3H). LCMS-A: sp 6.665 min, m/z 381.1 [M+H] ⁺ .

Example 154: N- (4-chlorobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 154

A mixture of 5-ethyl-2-methoxybenzenesulfonyl chloride I112 (0.414 g, 1.77 mmol) and 4-chlorobenzo[ d ]isoxazol-3-amine (0.225 g, 1.34 mmol) in pyridine (2.0 mL) was stirred at 30 °C for 40 h under nitrogen atmosphere. The reaction mixture was concentrated and then sonicated for 2 h using aqueous HCl (5%) and the resulting precipitate was collected. The residue was purified by silica gel column chromatography (0-100% ethyl acetate/petroleum gasoline, 40-60 °C) to afford the title compound as two fractions (A and B) in a combined yield of 0.060 g, 12% yield.

Fraction A : yield 0.038 g. ¹ H NMR (400 MHz, acetone -d ₆ ) δ 8.88 (br s, 1H), 7.75 (d, J= 2.3 Hz, 1H), 7.66 (dd, J= 8.5, 7.6 Hz, 1H), 7.57 (dd, J= 8.5, 0.6 Hz, 1H), 7.49 (dd, J= 8.5, 2.3 Hz, 1H), 7.42 (dd, J= 7.6, 0.6 Hz, 1H), 7.14 (d, J= 8.5 Hz, 1H), 3.91 (s, 3H), 2.66 (q, J= 7.6 Hz, 2H), 1.21 (t, J= 7.6 Hz, 3H). LCMS-B: sp 3.766 min; m/z 367.1/369.1 [M+H] ⁺ .

Fraction B: yield 0.021 g. ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.88 (br s, 1H), 7.75 (d, J = 2.3 Hz, 1H), 7.65 (dd, J = 8.5, 7.6 Hz, 1H), 7.59-7.55 (m, 1H), 7.49 (dd, J = 8.5, 2.3 Hz, 1H), 7.44-7.40 (m, 1H), 7.14 (d, J = 8.5 Hz, 1H), 3.90 (s, 3H), 2.66 (q, J = 7.6 Hz, 2H), 1.21 (t, J = 7.6 Hz, 3H). LCMS-B: sp. 3.755 min; m/z 367.1/369.1 [M+H] ⁺

Example 155: N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 155

A mixture of 4-chlorobenzo[ d ]isoxazol-3-amine (0.034 g, 0.200 mmol) and 2-methoxybenzenesulfonyl chloride (0.092 g, 0.450 mmol) in pyridine (1.0 mL) and triethylamine (0.1 mL) was stirred at room temperature for 16 h. The reaction mixture was concentrated, diluted with 5% aqueous HCl (1 mL), and sonicated for a minimum of 30 min. The resulting precipitate was collected by filtration and purified by preparative HPLC with mass separation to afford the title compound. ¹ H NMR (400 MHz, acetone- d ₆ ) δ 7.94-7.91 (dd, J= 7.8, 1.7 Hz, 1H), 7.69-7.63 (m, 2H), 7.60-7.57 (dd, J= 8.5, 0.7 Hz, 1H), 7.44-7.42 (dd, J= 7.6, 0.7 Hz, 1H), 7.25-7.22 (m, 1H), 7.16-7.11 (m, 1H), 3.94-3.94 (s, 3H). HPLC-MS: sp temp 6.02 min; m/z 339.16/341.18 [M+H] ⁺ .

Example 156: N- (4-fluorobenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 156

A mixture of 4-fluorobenzo[ d ]isoxazol-3-amine (0.032 g, 0.21 mmol) and 2-methoxybenzenesulfonyl chloride (0.109 g, 0.529 mmol) in pyridine (1.0 mL) and triethylamine (0.1 mL) was stirred at room temperature for 16 h. The reaction mixture was concentrated, diluted with 5% aqueous HCl (1 mL), and sonicated for a minimum of 30 min. The resulting precipitate was collected by filtration and purified by preparative HPLC with mass separation to afford the title compound. ¹ H NMR (400 MHz, acetone- d ₆ ) δ 9.73-9.45 (br s, 1H), 7.92-7.88 (dd, J= 7.9, 1.7 Hz, 1H), 7.70-7.61 (m, 2H), 7.44-7.40 (d, J= 8.5 Hz, 1H), 7.25-7.21 (d, J= 8.3 Hz, 1H), 7.13-7.07 (m, 2H), 3.95-3.91 (s, 3H). HPLC-MS: sp temp 5.72 min; m/z 323.16 [M+H] ⁺ .

Example 157: N- (6-bromobenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 157

A mixture of 6-bromobenzo[ d ]isoxazol-3-amine I75 (0.039 g, 0.180 mmol) and 2-methoxybenzenesulfonyl chloride (0.101 g, 0.490 mmol) in pyridine (1.0 mL) and triethylamine (0.1 mL) was stirred at room temperature for 16 h. The reaction mixture was concentrated, diluted with 5% aqueous HCl (1 mL), and sonicated for a minimum of 30 min. The resulting precipitate was collected by filtration and purified by preparative HPLC with mass separation to afford the title compound. ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.06-8.02 (dd, J= 8.6, 0.5 Hz, 1H), 7.90-7.87 (dd, J= 7.9, 1.7 Hz, 1H), 7.85-7.83 (dd, J= 1.6, 0.5 Hz, 1H), 7.64-7.59 (ddd, J= 8.4, 7.4, 1.8 Hz, 1H), 7.58-7.54 (dd, J= 8.6, 1.6 Hz, 1H), 7.21-7.18 (dd, J= 8.4, 0.8 Hz, 1H), 7.10-7.05 (m, 1H), 3.88-3.85 (s, 3H). HPLC-MS: time beat 6.32 min; m/z 383.1/385.2 [M+H] ⁺ .

Example 158: N- (6-chlorobenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 158

A mixture of 6-chlorobenzo[ d ]isoxazol-3-amine (0.033 g, 0.200 mmol) and 2-methoxybenzenesulfonyl chloride (0.095 g, 0.460 mmol) was stirred in pyridine (1.0 mL) and triethylamine (0.1 mL) at room temperature for 16 h. The reaction mixture was concentrated and diluted with 5% aqueous HCl (1 mL) and sonicated for a minimum of 30 min. The resulting precipitate was collected by filtration and purified by preparative HPLC with mass separation to afford the title compound. ¹ H NMR (400 MHz, acetone- d ₆ ) δ 8.12-8.07 (dd, J= 8.6, 0.5 Hz, 1H), 7.91-7.87 (dd, J= 7.9, 1.7 Hz, 1H), 7.68-7.65 (dd, J= 1.7, 0.5 Hz, 1H), 7.64-7.58 (ddd, J= 8.4, 7.4, 1.8 Hz, 1H), 7.44-7.39 (dd, J= 8.6, 1.7 Hz, 1H), 7.21-7.17 (m, 1H), 7.10-7.05 (m, 1H), 3.88-3.86 (s, 3H). HPLC-MS: sp 6.26 min; m/z 339.16/341.18 [M+H]+.

Example 159: N- (4-chlorobenzo[ d ]isoxazol-3-yl)isoquinoline-8-sulfonamide 159

A solution of 4-chlorobenzo[ d ]isoxazol-3-amine (0.050 g, 0.30 mmol) in anhydrous THF (2 mL) was cooled to -78 °C under nitrogen. A solution of lithium bis(trimethylsilyl)amide (1.0 M in THF, 0.59 mL, 0.59 mmol) was carefully added and the mixture was stirred at 0 °C for 1 h. The mixture was cooled to -78 °C, a solution of 8-isoquinolinesulfonyl chloride (0.068 g, 0.30 mmol) in anhydrous THF (1 mL) was added and the mixture was allowed to warm to room temperature. After stirring for 3 h, TLC showed only the presence of starting material. The mixture was cooled to -78 °C, sodium hydride (60% dispersion in mineral oil, 0.059 g, 1.5 mmol) was added, the mixture was allowed to warm back to room temperature and stirred overnight. Water (10 mL) was added and the pH was adjusted to ~3 using aq. HCl (2 M). The aqueous phase was extracted with DCM (3 x 20 mL), the organic layers were combined, dried (MgSO ₄ ) and the solvent was removed in vacuo. The solid residue was purified by column chromatography (Biotage Isolera, 12 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline 40-60 °C then 0-40% MeOH in EtOAc) to give the title compound as a white solid (0.026 g, 24%). ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.31 (s, 1H), 8.53 (d, J= 5.8 Hz, 1H), 8.28-8.21 (m, 1H), 8.09 (d, J= 8.2 Hz, 1H), 7.92 (d, J= 5.7 Hz, 1H), 7.89-7.82 (m, 1H), 7.39 (t, J= 7.9 Hz, 1H), 7.31 (d, J= 8.3 Hz, 1H), 7.17 (d, J= 7.5 Hz, 1H). LCMS-A: sp 5.43 min; m/z 360.1 [M+H] ⁺ .

Example 160: N -(7-iodo-4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 160

a) 7-iodo-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine A5

A portion of 4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I9 (0.121 g, 0.581 mmol) was dissolved in N,N -dimethylformamide (2 mL) and N -iodosuccinimide (0.131 g, 0.581 mmol) was added. After complete addition, the reaction mixture was heated at 50 °C for 2 h. At the end of this period, the reaction mixture was poured onto ice and then diluted with EtOAc (15 mL). The resulting mixture was washed with H ₂ O (3 × 8 mL) and brine (8 mL), dried over Na ₂ SO ₄ and filtered. The volatiles were removed under reduced pressure and the residue was purified twice by column chromatography (12 g SiO ₂ cartridge, 0-35% EtOAc in petroleum gasoline 40-60 °C, then 12 g SiO ₂ cartridge, 0-25% EtOAc in petroleum gasoline 40-60 °C) to give the title compound (0.038 g, 20% yield) as an off-white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.78 (s, 1H), 4.73 (s, 2H), 4.54 (s, 2H), 3.96 (s, 3H), 3.50 (s, 3H). LCMS-A: sp. 3.26 min, m/z 334.7 [M+H] ⁺ .

b) N-(7-iodo-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 160

A solution of 7-iodo-4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine A5 (0.024 g, 0.099 mmol) and 2,6-dimethoxybenzenesulfonyl chloride I111 (0.023 g, 0.099 mmol) in pyridine (0.5 mL) was irradiated in a microwave reactor at 110 °C for 2 h. The reaction mixture was cooled to room temperature and loaded wet onto a silica gel cartridge. The residue was purified by column chromatography (12 g SiO ₂ cartridge, 0-70% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (0.032 g, 53% yield) as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.38 (t, J =8.5 Hz, 1H), 6.86 (s, 1H), 6.58 (d, J =8.5 Hz, 2H), 4.52 (s, 2H), 4.04 (s, 3H), 3.88 (s, 6H), 3.51 (c, 3H). ZHMS-A: vr. beat 5.86 min, m/z 534.6 [M+H] ⁺ .

Example 161: N- (7-chloro-4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 161

a) 7-Chloro-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-amine A6

4-Methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I9 (0.150 g, 0.720 mmol) was dissolved in N,N- dimethylformamide (2 mL) and N- chlorosuccinimide (96 mg, 0.72 mmol) was added. After complete addition, the reaction mixture was heated at 50 °C for 2 h. At the end of this period, the reaction mixture was poured onto ice and diluted with EtOAc (15 mL). The resulting mixture was washed with H ₂ O (3 × 8 mL) and brine (8 mL), dried over Na ₂ SO ₄ and filtered. The volatiles were removed under reduced pressure and the residue was purified by column chromatography (12 g SiO ₂ cartridge, 0-40% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (0.0240 g, 14% yield) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 6.77 (s, 1H), 4.63 (d, J= 0.6 Hz, 2H), 3.97 (s, 3H), 3.49 (s, 3H).

b) N-(7-chloro-4-methoxy-6-(methoxymethyl)benzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 161

A solution of 7-chloro-4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine A6 (0.024 g, 0.099 mmol) and 2,6-dimethoxybenzenesulfonyl chloride I111 (0.023 g, 0.099 mmol) in pyridine (0.5 mL) was irradiated in a microwave reactor at 110 °C for 2 h. The reaction mixture was cooled to room temperature and loaded wet onto a silica gel cartridge. The residue was purified by column chromatography (12 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline 40-60 °C) to give the title compound (0.0094 g, 21% yield) as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 8.20 (s, 1H), 7.39 (t, J= 8.5 Hz, 1H), 6.85 (s, 1H), 6.59 (d, J= 8.5 Hz, 2H), 4.61 (d, J= 0.6 Hz, 2H), 4.04 (s, 3H), 3.88 (s, 6H), 3.49 (s, 3H). ZHMS-F: vr. beat 6.39 min, m/z 442.8 [M+H] ⁺ .

Example 162: 5-methoxy- N- (4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)quinoline-8-sulfonamide 162

A solution of 4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I9 (0.0500 g, 0.240 mmol) and 5-methoxyquinoline-8-sulfonyl chloride (0.0619 g, 0.240 mmol) in pyridine (0.500 mL) was irradiated in a microwave reactor at 110 °C for 2 h. The reaction mixture was cooled to room temperature and added to DCM (10 mL). The organic layer was washed with 0.5 M HCl (10 mL) and the layers were separated using a phase separation cartridge. The collected organic layers were dried in vacuo and the residue was purified by column chromatography (SiO2 cartridge ₁₂ g, 0-80% EtOAc in petroleum gasoline 40-60 °C) to give the title compound (0.0110 g, 11% yield) as an off-white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 8.98 (dd, J= 4.3, 1.8 Hz, 1H), 8.64-8.53 (m, 2H), 7.45 (dd, J= 8.5, 4.3 Hz, 1H), 6.91 (d, J= 8.4 Hz, 1H), 6.87 (d, J= 0.9 Hz, 1H), 6.54 (s, 1H), 4.44 (s, 2H), 4.05 (s, 3H), 4.03 (s, 3H), 3.36 (s, 3H). ZHMS-B: vr. beat 3.49 min, m/z =429.8 [M+H] ⁺ .

Example 163: 2-hydroxy-6-methoxy- N- (4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 163

A solution of 4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I9 (0.0440 g, 0.211 mmol) and 2,6-dimethoxybenzenesulfonyl chloride I111 (0.0500 g, 0.211 mmol) in pyridine (0.500 mL) was irradiated in a microwave reactor at 120 °C for 2 h and then at 120 °C for 1 h. The reaction mixture was cooled to room temperature and added to DCM (10 mL). The organic layers were washed with 1 M HCl (2 × 10 mL) and then dried over MgSO ₄ . The crude material was purified twice by silica gel chromatography (24 g SiO ₂ cartridge, 0-85% EtOAc in petroleum gasoline 40-60 °C, then 12 g SiO ₂ cartridge, 0-75% EtOAc in petroleum gasoline 40-60 °C) to give the title compound (1.5 mg). ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.62 (s, 1H), 8.14 (s, 1H), 7.33 (t, J= 8.4 Hz, 1H), 6.99 (k, J= 0.9 Hz, 1H), 6.70-6.61 (m, 2H), 6.37 (dd, J= 8.3, 1.0 Hz, 1H), 4.51 (s, 1H), 4.03 (s, 3H), 3.87 (s, 3H), 3.42 (s, 3H). ZHMS-A: vr. beat 3.54 min, m/z 394.8 [M+H] ⁺

Example 164: 6-methoxy- N -(6-(methoxymethyl)-5-methylbenzo[ d ]isoxazol-3-yl)pyridin-3-sulfonamide 164

A solution of 6-methoxypyridine-3-sulfonyl chloride (0.0540 g, 0.260 mmol) and 6-(methoxymethyl)-5-methylbenzo[ d ]isoxazol-3-amine I4 (0.050 g, 0.26 mmol) in pyridine (0.500 mL) was irradiated in a microwave reactor at 120 °C for 2 h. The reaction mixture was cooled to room temperature, then poured into DCM and washed (2 times) with 1 M HCl. The organic layer was dried in vacuo , then loaded wet onto silica gel and the product was purified by column chromatography ( ₂₄ g SiO2 cartridge, 0-80% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (15.6 mg, 17% yield) as a colorless oil. ^1H NMR (400 MHz, CDCl ₃ ) δ 8.63 (dd, J= 0.71, 2.62 Hz, 1H), 7.99-7.89 (m, 2H), 7.72 (s, 1H), 7.55 (s, 1H), 6.75 (dd, J= 0.71, 8.94 Hz, 1H), 4.54 (s, 2H), 3.95 (s, 3H), 3.50 (s, 3H), 2.39 (s, 3H). ZHMS-F: vr. beat 6.39 min, m/z 348.1 [M+H] ⁺ .

Example 165: N- (6-(methoxymethyl)-5-methylbenzo[ d ]isoxazol-3-yl)pyridin-3-sulfonamide 165

A solution of pyridine-3-sulfonyl chloride (0.0462 g, 0.260 mmol) and 6-(methoxymethyl)-5-methylbenzo[ d ]isoxazol-3-amine I4 (0.050 g, 0.26 mmol) in pyridine (0.500 mL) was irradiated in a microwave reactor at 120 °C for 2 h. The reaction mixture was cooled to room temperature and then loaded wet onto silica gel and the product was purified by column chromatography (24 g SiO ₂ cartridge, 0-80% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (0.0220 g, 25% yield) as a yellow solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.09 (s, 1H), 8.83 (s, 1H), 8.18 (d, J = 8.09 Hz, 1H), 7.70 (s, 1H), 7.57 (s, 1H), 7.46 (s, 1H), 4.54 (s, 2H), 3.50 (s, 3H), 2.40 (s, 3H). LCMS-F: sp 6.12 min m/z 334.1 [M+H] ⁺ , 332.0 [MH] ^- .

Example 166: 2,4-dimethoxy- N- (6-(methoxymethyl)-5-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 166

A solution of 2,4-dimethoxybenzenesulfonyl chloride (0.052 g, 0.22 mmol) and 6-(methoxymethyl)-5-methylbenzo[ d ]isoxazol-3-amine I4 (0.042 g, 0.22 mmol) in pyridine (0.500 mL) was irradiated in a microwave reactor at 110 °C for 2 h. The resulting mixture was loaded onto silica gel and the product was purified by column chromatography (4 g SiO ₂ cartridge, 0-45% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (0.0413 g, 48% yield) as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.96 (s, 1H), 7.83 (s, 1H), 7.71 (d, J= 8.33 Hz, 1H), 7.47 (s, 1H), 6.48 (d, J= 1.82 Hz, 1H), 6.42 (d, J= 8.36 Hz, 1H), 4.50 (s, 2H), 3.95 (s, 3H), 3.80 (s, 3H), 3.47 (s, 3H), 2.37 (s, 3H). ZHMS-A: vr. beat 5.78 min, m/z =392.8 [M+H] ⁺ , 414.7 [M+Na] ⁺ .

Example 167: N- (6-(hydroxymethyl)benzo[ d ]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide 167

a) Methyl 3-aminobenzo[d]isoxazole-6-carboxylate A9

To a solution of ethanehydroxamic acid (0.629 g, 8.37 mmol) in DMF (5 mL) was added potassium 2-methylpropane - 2-olate (0.94 g, 8.4 mmol) and the reaction mixture was stirred for 30 min. Methyl 4-cyano-3-fluorobenzoate (1.0 g, 5.6 mmol) was added followed by DMF (2 mL) and the reaction mixture was stirred for an additional 2 h at room temperature. The reaction mixture was diluted with ethyl acetate (50 mL) and water (50 mL), the aqueous layer was extracted with ethyl acetate, the combined organic layers were washed with water, dried, filtered and concentrated. The crude material was purified by silica gel chromatography (12 g SiO ₂ cartridge, 0-50% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (0.55 g, 51% yield) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.12 (d, J = 1.02 Hz, 1H), 7.96 (dd, J = 1.24, 8.29 Hz, 1H), 7.59 (dd, J = 0.77, 8.25 Hz, 1H), 3.98 (s, 3H). LCMS: sp 2.97 min, m/z 193.0 [M+H] ⁺ .

b) Methyl 3-((2,4-dimethoxyphenyl)sulfonamido)benzo[d]isoxazole-6-carboxylate A10

A solution of 2,4-dimethoxybenzene-1-sulfonyl chloride (0.67 g, 2.8 mmol) and methyl 3-aminobenzo[ d ]isoxazole-6-carboxylate A9 (0.55 g, 2.8 mmol) in pyridine (4 mL) was irradiated in a microwave reactor at 130 °C for 3 h. The reaction mixture was cooled to room temperature and diluted with DCM (40 mL). The organic layers were washed with 1 M HCl (40 mL), the aqueous layer was extracted again with DCM (2 × 40 mL). The combined organic layers were dried in vacuo and the residue was purified twice by column chromatography ( ₂₄ g SiO2 cartridge, 0-35% EtOAc in petroleum gasoline, 40-60 °C) to give two crops of the title compound (0.369 g crude and 0.0310 g yield, 2.8% with >95% purity) as white solids. ^1H NMR (400 MHz, methanol - _d4 ) δ 8.13-8.06 (m, 2H), 7.97 (dd, J= 1.25, 8.47 Hz, 1H), 7.86-7.80 (m, 1H), 6.58 (dk, J= 2.29, 4.60 Hz, 2H), 3.95 (s, 3H), 3.83 (s, 3H), 3.79 (s, 3H). JHMS: vr. beat 3.26 min, m/z 392.8 [M+H] ⁺ , 415.8 [M+Na] ⁺ .

c) N-(6-(hydroxymethyl)benzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide 167

To a suspension of lithium aluminum hydride powder (0.0758 g, 2.00 mmol) in anhydrous THF (4 mL) under nitrogen atmosphere was added a solution of methyl 3-((2,4-dimethoxyphenyl)sulfonamido)benzo[ d ]isoxazole-6-carboxylate A10 (crude, 0.392 g, 0.500 mmol) in THF (8 mL). The mixture was stirred overnight at room temperature. The reaction was quenched under nitrogen atmosphere by dropwise addition of wet THF followed by 1 mL of water. After gas evolution ceased, 0.5 M aqueous HCl solution was added and the aqueous layer was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with water, brine, dried over MgSO ₄ and the solvent was removed in vacuo. The crude residue was purified by column chromatography ( ₂₄ g SiO2 cartridge, 0-100% EtOAc in petroleum gasoline 40-60 °C) to give the title compound (0.191 g, 100% yield) as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 8.08 (dd, J= 0.78, 8.30 Hz, 1H), 7.77 (s, 1H), 7.69 (d, J= 8.82 Hz, 1H), 7.47 (t, J= 1.03 Hz, 1H), 7.30 (dd, J= 1.34, 8.29 Hz, 1H), 6.49 (d, J= 2.26 Hz, 1H), 6.42 (dd, J= 2.30, 8.79 Hz, 1H), 4.84 (s, 2H), 3.98 (s, 3H), 3.80 (s, 3H). ZHMS-B: vr. beat 3.02 min, m/z 364.8 [M+H] ⁺ , 386.8 [M+Na] ⁺ .

Example 168: 3-((5-methoxyquinoline)-8-sulfonamido)-5-methylbenzo[ d ]isoxazole-6-carboxamide 168

a) Methyl 3-amino-5-methylbenzo[d]isoxazole-6-carboxylate A11

To a solution of ethanehydroxamic acid (0.126 g, 1.69 mmol) in DMF (2 mL) was added potassium 2-methylpropane - 2-olate (0.19 g, 1.7 mmol) and the reaction mixture was stirred for 30 min. Methyl 4-cyano-5-fluoro-2-methylbenzoate (0.22 g, 1.1 mmol) was added followed by DMF (3 mL) and the reaction mixture was stirred for an additional 2 h at 40 °C. The reaction mixture was diluted with ethyl acetate (50 mL) and water (50 mL). The aqueous layer was extracted with ethyl acetate and the combined organic layers were washed with water, dried, filtered and concentrated. The crude material was purified by silica gel chromatography (12 g SiO ₂ cartridge, 0-50% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (0.11 g, 48% yield) as a white solid. ¹ H NMR (400 MHz, methanol -d ₄ ) δ 7.85 (s, 1H), 7.64 (t, J = 0.79 Hz, 1H), 3.92 (s, 3H), 2.62 (d, J = 0.85 Hz, 3H). LCMS: sp 3.02 min, m/z 207.0 [M+H] ⁺ .

b) Methyl 3-((5-methoxyquinoline)-8-sulfonamido)-5-methylbenzo[d]isoxazole-6-carboxylate hydrochloride salt A12

A solution of 5-methoxyquinoline-8-sulfonyl chloride (0.12 g, 0.48 mmol) and methyl 3-amino-5-methylbenzo[ d ]isoxazole-6-carboxylate A11 (0.10 g, 0.48 mmol) in pyridine (3 mL) was irradiated in a microwave at 110 °C for 2 h. The reaction mixture was irradiated in a microwave at 110 °C for an additional 1.5 h. The resulting mixture was loaded onto silica gel and the product was purified by column chromatography (4 g SiO ₂ cartridge, 0-45% EtOAc in petroleum gasoline, 40-60 °C). The product was further purified by solid phase extraction (1 g, Si-amine, 3 void volumes of MeOH, followed by 4 void volumes of methanolic HCl). The acidic eluates were collected and dried in vacuo to give the title compound (17.7 mg, 7.9% yield) as a pale yellow solid. ¹ H NMR (400 MHz, methanol -d ₄ ) δ 9.40-9.33 (m, 2H), 8.77 (d, J = 8.66 Hz, 1H), 8.14-8.05 (m, 1H), 7.91 (s, 1H), 7.80-7.73 (m, 1H), 7.47 (d, J = 8.73 Hz, 1H), 4.23 (s, 3H), 3.91 (s, 3H), 2.62 (s, 3H). LCMS: sp. 3.35 min, m/z 427.8 [M+H] ⁺ .

c) 3-((5-Methoxyquinoline)-8-sulfonamido)-5-methylbenzo[d]isoxazole-6-carboxamide 168

To a 15 mL pressure test tube equipped with a magnetic stirrer under nitrogen atmosphere were added methyl 3-((5-methoxyquinoline)-8-sulfonamido)-5-methylbenzo[ d ]isoxazole-6-carboxylate hydrochloride salt A12 (15.0 mg, 0.0323 mmol), ammonia solution (2.0 M in methanol, 0.50 mL, 1.0 mmol), and calcium dichloride (3.59 mg, 0.0323 mmol). The reaction vessel was sealed and heated at 80 °C for 3 days. The solvent was removed in a stream of air, and ammonia solution (7.0 M in methanol, 0.50 mL, 3.5 mmol) and calcium dichloride (3.6 mg, 0.032 mmol) were added. The reaction vessel was sealed and heated at 80 °C for 24 h. The reaction mixture was cooled to room temperature and the solvent was removed in vacuo. The compound was purified by column chromatography (4 g SiO ₂ cartridge, 0-100% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (0.00180 g, 13% yield) as a white solid. ¹ H NMR (400 MHz, methanol- d ₄ ) δ 9.01 (dd, J = 1.78, 4.24 Hz, 1H), 8.65 (dd, J = 1.78, 8.52 Hz, 1H), 8.43 (d, J = 8.47 Hz, 1H), 7.73 (d, J = 12.46 Hz, 2H), 7.57 (dd, J = 4.28, 8.54 Hz, 1H), 7.08 (d, J = 8.50 Hz, 1H), 4.09 (s, 3H), 2.57 (s, 3H). LCMS-B: sp 3.15 min, m/z 413.8 [M+H] ⁺ .

Example 169: N- (6-cyano-5-methylbenzo[ d ]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide 169

a) 3-Amino-5-methylbenzo[d]isoxazole-6-carbonitrile A13

To a solution of ethanehydroxamic acid (0.176 g, 2.34 mmol) in DMF (5 mL) was added potassium 2- methylpropane 2-olate (0.26 g, 2.3 mmol) and the reaction mixture was stirred for 30 min. 2-Fluoro-5-methyl-terephthalonitrile (0.25 g, 1.6 mmol) was added followed by DMF (3 mL) and the reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted with ethyl acetate (50 mL) and water (50 mL), the aqueous layer was extracted with ethyl acetate, the combined organic layers were washed with water, dried, filtered and concentrated. The crude material was purified by silica gel chromatography (12 g SiO ₂ cartridge, 0-50% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (0.16 g, 59% yield) as an off-white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.72 (s, 1H), 7.47 (t, J = 0.86 Hz, 1H), 2.65 (d, J = 0.86 Hz, 3H). LCMS-B: sp 2.90 min, m/z 174.0 [M+H] ⁺ .

b) N-(6-cyano-5-methylbenzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide 169

A solution of 2,4-dimethoxybenzene-1-sulfonyl chloride (0.22 g, 0.92 mmol) and 3-amino-5-methyl-1,2-benzoxazole-6-carbonitrile A13 (0.16 g, 0.92 mmol) in pyridine (2.5 mL) was irradiated in a microwave reactor at 130 °C for 2 h. The reaction was carried out at room temperature for 50 min, and then the reaction mixture was irradiated in a microwave reactor at 130 °C for an additional 2 h. The reaction mixture was cooled to room temperature and diluted with DCM (40 mL). The organic layers were washed with 1 M HCl (40 mL), the aqueous layer was extracted twice with DCM (2 × 40 mL). The combined organic layers were dried in vacuo, the residue was loaded onto silica gel, and the product was purified by column chromatography ( ₂₄ g SiO2 cartridge, 0-45% EtOAc in petroleum gasoline, 40-60 °C) to give a yellow solid. The solid was dissolved in warm MeOH and DCM and purified by solid phase extraction (1 g Si-amine, 3 void volumes of MeOH, followed by 3 void volumes of ~1.25 M methanolic ammonia). The acidic eluate was dried in vacuo to give a white solid. The solid was taken up in MeOH and the MeOH was removed in vacuo (repeated 3 times). The residue was re-purified by column chromatography ( ₂₄ g SiO2 cartridge, 45% EtOAc in petroleum gasoline, 40-60 °C) to give two crops of the title compound (22 and 78 mg, total weight 100 mg, yield 29%) as an off-white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.09 (s, 1H), 7.82 (s, 1H), 7.72 (s, 1H), 7.66 (d, J= 8.80 Hz, 1H), 6.52 (d, J= 2.25 Hz, 1H), 6.44 (dd, J= 2.24, 8.85 Hz, 1H), 3.98 (s, 3H), 3.82 (s, 3H), 2.67 (s, 3H). LCMS-B: sp 3.30 min, m/z = 373.8 [M+H] ⁺ , 371.9 [MH] ^- .

Example 170: 2,4-dimethoxy- N- (5-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 170

A solution of 2,4-dimethoxybenzene-1-sulfonyl chloride (0.0799 g, 0.337 mmol) and 5-methylbenzo[ d ]isoxazol-3-amine I60 (0.050 g, 0.34 mmol) in pyridine (1 mL) was irradiated in a microwave reactor at 110 °C for 2 h. The resulting mixture was loaded onto silica gel and the product was purified by column chromatography (12 g SiO ₂ cartridge, 0-45% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (55.0 mg, 42% yield) as a white solid. ^1H NMR (400 MHz, CDCl ₃ ) δ 7.86-7.82 (m, 1H), 7.72 (d, J= 8.81 Hz, 1H), 7.37-7.28 (m, 2H), 6.49 (d, J= 2.26 Hz, 1H), 6.43 (dd, J= 2.29, 8.82 Hz, 1H), 3.96 (s, 3H), 3.80 (s, 3H), 2.47 (s, 3H). ZHMS-A: vr. beat 5.66 min, m/z 348.8 [M+H] ⁺ , 347.1 [MH] ^- .

Example 171: N- (6-bromo-5-methylbenzo[ d ]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide 171

a) 6-Bromo-5-methylbenzo[d]isoxazol-3-amine A14

To a solution of ethanehydroxamic acid (0.263 g, 3.50 mmol) in N,N- dimethylformamide (5 mL) was added t -BuOK (393 mg, 3.50 mmol) and the reaction mixture was stirred for 30 min. To the reaction mixture was added 4-bromo-2-fluoro-5-methylbenzonitrile (0.50 g, 2.3 mmol) and stirred for additional 2 h at room temperature. The reaction mixture was diluted with ethyl acetate (50 mL) and water (50 mL), the aqueous layer was extracted using ethyl acetate, the combined organic layers were washed with water, dried, filtered and concentrated. The crude material was purified by silica gel chromatography (12 g SiO ₂ cartridge, 0-50% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (0.30 g, 56% yield) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.68 (s, 1H), 7.37 (s, 1H), 4.35 (br s, 2H), 2.49 (s, 3H). LCMS-B: sp 3.18 min, m/z 229.8 [M+H] ⁺ .

b) N-(6-bromo-5-methylbenzo[d]isoxazol-3-yl)-2,4-dimethoxybenzenesulfonamide 171

A solution of 2,4-dimethoxybenzenesulfonyl chloride (0.052 g, 0.22 mmol) and 6-bromo-5-methylbenzo[ d ]isoxazol-3-amine A14 (0.050 g, 0.22 mmol) in pyridine (1 mL) was irradiated twice in a microwave reactor at 110 °C for 2 h and then at 130 °C for 2 h. The resulting mixture was loaded onto silica gel and the product was purified by column chromatography (4 g SiO ₂ cartridge, 0-45% EtOAc in petroleum gasoline, 40-60 °C) to give the title compound (quantitative yield 102 mg) as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.98-7.94 (m, 1H), 7.70-7.65 (m, 2H), 6.50 (d, J= 2.24 Hz, 1H), 6.43 (dd, J= 2.26, 8.82 Hz, 1H), 3.97 (s, 3H), 3.81 (s, 3H), 2.51 (d, J= 0.87 Hz, 4H). LCMS-A: sp 6.08 min, m/z 426.9 [M+H] ⁺ .

Example 172: N -(6-(ethoxymethyl)-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 172

To a solution of 6-(ethoxymethyl)-4-methoxybenzo[ d ]isoxazol-3-amine I21 (250 mg, 1.13 mmol) in anhydrous THF (25 mL) at -78 °C under N ₂ was added dropwise LiHMDS (1 M solution in THF, 4.5 mL, 4.5 mmol) and the mixture was stirred at -78 °C for 2 h. Then a solution of 2,6-dimethoxybenzenesulfonyl chloride I111 (400 mg, 1.69 mmol) in anhydrous THF (2 mL) was added dropwise and the mixture was allowed to warm to rt and stirred overnight. The mixture was acidified to pH 4-5 with 2 M aqueous HCl and extracted with EtOAc. The combined organic extracts were washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=100/1) to give the title compound (170 mg, 96% purity) as a white solid. Further purification by prep. HPLC afforded the title compound (60 mg, 100% purity, 13% yield). LCMS-C: _sp 2.08 min; m/z 423.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.53 (s, 1H), 7.51 (t, J =8.4 Hz, 1H), 7.09 (s, 1H), 6.78-6.76 (m, 3H), 4.55 (s, 2H), 3.91 (s, 3H), 3.77 (s, 6H), 3.54 (k, J =6.8 Hz, 2H), 1.19 (t, J =6.8 Hz, 3H).

Example 173: 2,6-dimethoxy- N -(4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 173

To a solution of 4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I9 (3.0 g, 14.4 mmol) in anhydrous THF (200 mL) at -78 °C under N ₂ was added dropwise LiHMDS (1 M solution in THF, 43.2 mL, 43.2 mmol) and the mixture was stirred at -78 °C for 2 h. Then a solution of 2,6-dimethoxybenzenesulfonyl chloride I111 (5.1 g, 21.6 mmol) in anhydrous THF (10 mL) was added dropwise and the mixture was allowed to warm to rt and stirred overnight. The mixture was acidified to pH 4-5 with 2 M aqueous HCl and extracted with EtOAc. The combined organic extracts were washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The reaction was repeated using 4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-amine I9 (2.0 g, 9.6 mmol) in 150 mL THF, the two crops were combined and purified by column chromatography (pet. ether/EtOAc=8/1 to 2/1) to give the title compound (4.1 g, 42%) as a white solid. LCMS-C: _sp 1.96 min; m/z 409.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.58 (s, 1H), 7.52 (t, J = 8.4 Hz, 1H), 7.09 (s, 1H), 6.78 (d, J = 8.4 Hz, 2H), 6.76 (s, 1H), 4.51 (s, 2H), 3.91 (s, 3H), 3.77 (s, 6H), 3.33 (s, 3H).

Example 174: 2,6-dimethoxy- N -(4-methoxy-6-phenylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 174

To a solution of 4-methoxy-6-phenylbenzo[ d ]isoxazol-3-amine I17 (2.5 g, 10.4 mmol) in anhydrous THF (60 mL) at -78 °C under N ₂ atmosphere was added dropwise LiHMDS (1 M solution in THF, 31.0 mL, 31.0 mmol) and the mixture was stirred at -78 °C for 2 h. Then, a solution of 2,6-dimethoxybenzenesulfonyl chloride I111 (3.7 g, 15.6 mmol) in anhydrous THF (20 mL) was added dropwise and the mixture was allowed to warm to 0 °C and stirred overnight. Water was added and the mixture was washed with EtOAc (50 mL × 2). The aqueous layer was acidified to pH 3 with 1 M aqueous HCl solution and extracted with EtOAc (50 mL × 3). The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=300/1) and further purified by column chromatography (DCM/MeOH=200/1) to give the title compound (1.5 g, 33%) as a white solid. ¹ H NMR (400 MHz, DMSO _-d6 ) δ 9.61 (s, 1H), 7.80 (d, J= 7.2 Hz, 2H), 7.50-7.44 (m, 5H), 7.09 (s, 1H), 6.80 (d, J= 8.8 Hz, 2H), 4.02 (s, 3H), 3.79 (s, 6H); JHMS-C: vr. _beat 2.46 min; m/z 441.0 [M+H] ⁺ .

Example 175: 3-chloro-2,6-dimethoxy- N -(4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 175

To a solution of 2,6-dimethoxy- N- (4-methoxy-6-(methoxymethyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 173 (50 mg, 0.123 mmol) in DMF (10 mL) was added NCS (14 mg, 0.123 mmol), and the mixture was heated at 50 °C for 2 h. Then, the mixture was diluted with EtOAc (150 mL) and washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (DCM/MeOH=120/1) to give the title compound (15 mg, 27%) as a white solid. LCMS-C: _sp 2.21 min; m/z 441.0 [M+H] ⁺ . ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.56 (d, J = 9.1 Hz, 1H), 7.01 (s, 1H), 6.91 (d, J = 9.1 Hz, 1H), 6.76 (s, 1H), 4.55 (s, 2H), 3.99 (s, 6H), 3.76 (s, 3H), 3.41 (s, 3H).

Example 176: 2,6-dimethoxy- N -(4-methoxy-6-(2-methoxyphenyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 176

A mixture of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 89 (30 mg, 0.068 mmol), (2-methoxyphenyl)boronic acid (21 mg, 0.135 mmol), Pd( _PPh3 ) ₄ (9 mg, 0.007 mmol) and _{Na2CO3 (} 22 mg, 0.203 mmol) in 1,4-dioxane (4 mL) and water (1 mL) was heated at 100 °C under _N2 overnight. The mixture was diluted with water and extracted with EtOAc. The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified with prep. TLC (pet. ether/EtOAc=1/2) to give the title compound (10 mg, 31%) as a white solid. LCMS-C: _sp 2.36 min, m/z 471.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 9.60 (s, 1H), 7.51 (t, J =8.5 Hz, 1H), 7.44-7.37 (m, 2H), 7.22 (s, 1H), 7.16 (d, J =8.8 Hz, 1H), 7.05 (t, J= 7.4 Hz, 1H), 6.90 (s, 1H), 6.79 (d, J =8.6 Hz, 2H), 3.94 (s, 3H), 3.81 (s, 6H), 3.79 (s, 3H).

Example 177: 2,6-dimethoxy N -(4-methoxy-6-(3-methoxyphenyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 177

A mixture of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 89 (50 mg, 0.113 mmol), (3-methoxyphenyl)boronic acid (35 mg, 0.226 mmol), Pd( _PPh3 ) ₄ (14 mg, 0.011 mmol) and _{Na2CO3 (} 36 mg, 0.339 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was heated at 100 °C under _N2 overnight. The mixture was cooled to rt, adjusted to pH 4-5, diluted with water and extracted with EtOAc. The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=50/1) to give the title compound (8 mg, 15%) as a white solid. LCMS-C: _sp 2.35 min; m/z 471.0 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.63 (s, 1H), 7.51 (t, J =8.5 Hz, 1H), 7.46 (d, J =1.0 Hz, 1H), 7.41 (t, J =7.9 Hz, 1H), 7.37-7.29 (m, 2H), 7.07 (s, 1H), 7.03-6.98 (m, 1H), 6.79 (d, J =8.5 Hz, 2H), 4.02 (s, 3H), 3.84 (s, 3H), 3.79 (s, 6H).

Example 178: 5-ethyl-2-methoxy -N- (7-phenylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 178

A suspension of N- (7-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 137 (100 mg, 0.24 mmol), phenylboronic acid (60 mg, 0.48 mmol), Pd(dppf) _Cl2 (18 mg, 0.024 mmol) and _{K3PO4 3H2O} (260 mg, 0.97 mmol) in toluene (5 mL), isopropanol (2 mL) and water (5 mL) was heated at 100 °C under _N2 for 2 h. _The mixture was allowed to cool to rt, diluted with EtOAc and washed with water (25 mL×3). The organic layer was dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by preparative TLC (pet. ether/EtOAc=2/1) to give the title compound (55 mg, 55%) as a white solid. LCMS-D: _sp 3.06 min; m/z 409.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 8.08-8.02 (m, 1H), 7.89-7.78 (m, 3H), 7.72 (d, J= 2.3 Hz, 1H), 7.55-7.40 (m, 5H), 7.13-7.08 (m, 1H), 3.75 (s, 3H), 2.61 (k, J= 7.5 Hz, 2H), 1.15 (t, J= 7.5 Hz, 3H).

Example 179: 5-ethyl-2-methoxy -N- (6-phenylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 179

A mixture of N- (6-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 138 (120 mg, 0.3 mmol), Pd(dppf) _Cl2 (45 mg, 0.06 mmol), _{phenylboronic} acid (150 mg, 1.2 mmol) and _{K3PO4 3H2O} (399 mg, 1.5 mmol) in water (10 mL), toluene (10 mL) and isopropanol (5 mL) was heated at 85 °C under _N2 for 4 h. The mixture was allowed to cool to rt, diluted with water (200 mL) and extracted with diethyl ether (200 mL × 3). The combined organic extracts were washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 5/1) to give the title compound (60 mg, 50%) as a white solid. LCMS-D: _sp 3.10 min; m/z 409.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.7 (s, 1H), 8.14-8.08 (m, 1H), 7.87 (s, 1H), 7.79-7.74 (m, 2H), 7.73-7.67 (m, 2H), 7.54-7.39 (m, 4H), 7.12-7.07 (m, 1H), 3.74 (s, 3H), 2.61 (k, J= 7.5 Hz, 2H), 1.15 (t, J= 7.6 Hz, 3H).

Example 180: N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2,3-dihydrobenzofuran-7-sulfonamide 180

To a solution of 5-bromo -N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2,3-dihydrobenzofuran-7-sulfonamide 127 (100 mg, 0.23 mmol) in THF (10 mL) were added 10% Pd/C (20 mg) and KOAc (20 mg, 0.28 mmol) and the mixture was stirred at 40 °C under H ₂ for 2 h and then at rt overnight. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=30/1) to give the title compound (22 mg 27%) as a light yellow solid. LCMS-D: sp _temp 2.32 min; m/z 351.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.9 (s, 1H), 7.76-7.62 (m, 2H), 7.53-7.43 (m, 3H), 6.96 (t, J= 7.7 Hz, 1H), 4.50 (t, J= 8.8 Hz, 2H), 3.22 (t, J= 8.8 Hz, 2H).

Example 181: N- (4-chlorobenzo[ d ]isoxazol-3-yl)-5-ethyl-2,3-dihydrobenzofuran-7-sulfonamide 181

a) N-(4-Chlorobenzo[d]isoxazol-3-yl)-5-vinyl-2,3-dihydrobenzofuran-7-sulfonamide A15

To a solution of 5-bromo -N- (4-chlorobenzo[ d ]isoxazol-3-yl)-2,3-dihydrobenzofuran-7-sulfonamide 127 (200 mg, 0.47 mmol) in 1,4-dioxane (20 mL), EtOH ( ₁₀ mL) and _{H2O (10 mL) were added K2CO3 (} 206 mg, 1.86 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (140 mg, 0.93 mmol) and Pd( _PPh3 ) ₄ (54 mg, 0.047 mmol) and the mixture was heated at 90°C under _N2 overnight. The solvents were removed under reduced pressure and the residue was partitioned between DCM (50 mL), water (45 mL) and 2 M aq. HCl (5 mL). The layers were separated and the organic layer was washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (pet. ether/EtOAc=3/1) to give the title compound (120 mg, 70%) as a yellow solid. LCMS-D: _sp 0.40 min; m/z 377.0 [M+H] ⁺ .

b) N-(4-chlorobenzo[d]isoxazol-3-yl)-5-ethyl-2,3-dihydrobenzofuran-7-sulfonamide 181

To a solution of N- (4-chlorobenzo[ d ]isoxazol-3-yl)-5-vinyl-2,3-dihydrobenzofuran-7-sulfonamide A15 (120 mg, 0.32 mmol) in MeOH (15 mL) was added 10% Pd/C (24 mg) and the mixture was stirred at rt under H ₂ atmosphere overnight. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=30/1) to give the title compound (33 mg, 27%) as a white solid. LCMS-D: _sp 2.59 min; m/z 379.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.8 (s, 1H), 7.75-7.70 (m, 1H), 7.69-7.63 (m, 1H), 7.47-7.43 (m, 1H), 7.37 (s, 1H), 7.29 (s, 1H), 4.46 (t, J= 8.7 Hz, 2H), 3.18 (t, J= 8.7 Hz, 2H), 2.56 (k, J= 7.6 Hz, 2H), 1.13 (t, J= 7.6 Hz, 3H).

Example 182: N- (benzo[ d ]isoxazol-3-yl)-4-ethyl-2-methoxybenzenesulfonamide 182

a) N-(Benzo[d]isoxazol-3-yl)-2-methoxy-4-vinylbenzenesulfonamide A16

To a solution of N- (benzo[ d ]isoxazol-3-yl)-4-bromo-2-methoxybenzenesulfonamide 141 (200 mg, 0.52 mmol) in toluene (16 mL), water (8 mL) and isopropanol (8 mL) were added K ₂ CO ₃ (288 mg, 2.09 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (160 mg, 1.04 mmol) and Pd(PPh ₃ ) ₄ (60 mg, 0.052 mmol) and the mixture was heated at 90 °C under N ₂ for 2 h. The mixture was diluted with water (50 mL), 2 M aq. HCl (20 mL) and extracted with EtOAc (80 mL × 2). The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=5/1) to give the title compound (150 mg, 76%) as a light yellow solid. LCMS-D: _sp 2.47 min; m/z 331.0 [M+H] ⁺ , 353.0 [M+Na] ⁺ .

b) N-(Benzo[d]isoxazol-3-yl)-4-ethyl-2-methoxybenzenesulfonamide 182

To a solution of N- (benzo[ d ]isoxazol-3-yl)-2-methoxy-4-vinylbenzenesulfonamide A16 (80 mg, 0.24 mmol) in MeOH (10 mL) was added 10% Pd/C (16 mg) and the mixture was stirred at 25 °C overnight under H ₂ atmosphere. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by prep. TLC (pet. ether/EtOAc=5/1) to give the title compound (20 mg, 25%) as a light yellow solid. LCMS-D: _sp 2.55 min; m/z 333.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.7 (s, 1H), 8.07 (d, J= 8.0 Hz, 1H), 7.79 (d, J= 8.0 Hz, 1H), 7.66-7.55 (m, 2H), 7.44-7.31 (m, 1H), 7.01 (s, 1H), 6.96 (d, J= 8.4 Hz, 1H), 3.76 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H).

Example 183: N- (Benzo[ d ]isoxazol-3-yl)-3-methoxy-[1,1'-biphenyl]-4-sulfonamide 183

To a solution of N- (benzo[ d ]isoxazol-3-yl)-4-bromo-2-methoxybenzenesulfonamide 141 (100 mg, 0.26 mmol) in toluene (7 mL), water (7 mL) and isopropanol (2.5 mL) were added K ₂ CO ₃ (144 mg, 10 mmol), phenylboronic acid (64 mg, 0.52 mmol) and Pd(PPh ₃ ) ₄ (30 mg, 0.026 mmol) and the mixture was heated at 90 °C under N ₂ for 2 h. The mixture was diluted with water (50 mL), 2 M aq. HCl (10 mL) and extracted with EtOAc (70 mL × 2). The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=100/1 to 5/1) to give the title compound (55 mg, 46%) as a light yellow solid. LCMS-D: _sp 2.79 min; m/z 381.0 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.9 (s, 1H), 8.10 (d, J= 8.0 Hz, 1H), 7.98 (d, J= 8.0 Hz, 1H), 7.79-7.72 (m, 2H), 7.66-7.58 (m, 2H), 7.54-7.35 (m, 6H), 3.89 (s, 3H).

Example 184: 3-(5-ethyl-2-methoxyphenylsulfonamido) -N- methylbenzo[ d ]isoxazole-7-carboxamide 184

a) N-(7-Bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A17

To a solution of N- (7-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 137 (2.3 g, 5.5 mmol), (2,4-dimethoxyphenyl)methanol (1.4 g, 8.4 mmol) and _PPh3 (3.6 g, 14.2 mmol) in THF (400 mL) at 0 °C under _N2 was added DIAD (3.3 g, 16.4 mmol) and the mixture was stirred at rt over the weekend. The mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (pet. ether/EtOAc=5/1) to give the title compound (1.0 g, 32%) as a blue solid. LCMS-D: _sp 3.35 min; m/z 583.1/585.1 [M+Na] ⁺ .

b) Methyl 3-(N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxyphenylsulfonamido) benzo[d]isoxazole-7-carboxylate A18

To a solution of N- (7-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A17 (250 mg, 1.34 mmol) in MeOH (5 mL) and DMF (45 mL) were added _Et3N (675 mg, 6.68 mmol) and Pd(dppf) _Cl2 (98 mg, 0.13 mmol), and the mixture was heated at 80 °C under CO overnight. The solvent was removed under reduced pressure, the residue was diluted with EtOAc (50 mL), washed with water (50 mL×3), brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=4/1) to give the title compound (210 mg, 31%) as a white solid. LCMS-D: _sp 3.10 min; m/z 541.2 [M+H] ⁺ .

c) 3-(N-((2,4-Dimethoxybenzyl)oxy)-5-ethyl-2-methoxyphenylsulfonamido)-N-methylbenzo[d]isoxazole-7-carboxamide A19

A mixture of methyl 3-( N -((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxyphenylsulfonamido) _benzo [ d ]isoxazole-7-carboxylate A18 (20 mg, 0.037 mmol) and _CH3NH2 (33% solution in EtOH, 4 mL) was heated at 100 °C in a sealed tube for 30 min. The solvent was removed under reduced pressure to give the title compound (20 mg, 100%), which was used in the next step without further purification. LCMS-D: sp _temp 2.86 min; m/z 540.2 [M+H] ⁺ .

d) 3-(5-Ethyl-2-methoxyphenylsulfonamido)-N-methylbenzo[d]isoxazole-7-carboxamide 184

A mixture of 3-( N -((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxyphenylsulfonamido) -N- methylbenzo[ d ]isoxazole-7-carboxamide A19 (40 mg, 0.07 mmol) and TFA (2 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by preparative TLC to give the title compound (18 mg, 64%) as a white solid. LCMS-D: _sp 2.35 min; m/z 390.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.9 (s, 1H), 8.31-8.11 (m, 2H), 8.00-7.90 (m, 1H), 7.72 (s, 1H), 7.52-7.41 (m, 2H), 7.14-7.04 (m, 1H), 3.73 (s, 3H), 2.81 (s, 3H), 2.62 (k, J= 8.0, 7.6 Hz, 2H), 1.15 (t, J= 7.9 Hz, 3H).

Example 185: 3-(5-ethyl-2-methoxyphenylsulfonamido)- N, N -dimethylbenzo[ d ]isoxazole-7-carboxamide 185

a) Methyl 3-(5-ethyl-2-methoxyphenylsulfonamido)benzo[d]isoxazole-7-carboxylate A20

A mixture of methyl 3-( N -((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxyphenylsulfonamido)benzo[ d ]isoxazole-7-carboxylate A18 (150 mg, 0.28 mmol) and TFA (5 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=3/1) to give the title compound (80 mg, 74%) as a white solid. LCMS-D: _sp 2.63 min; m/z 391.1 [M+H] ⁺ , 413.1 [M+Na] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.9 (s, 1H), 8.39-8.32 (m, 1H), 8.19-8.12 (m, 1H), 7.72 (d, J= 2.2 Hz, 1H), 7.56-7.44 (m, 2H), 7.12-7.05 (m, 1H), 3.89 (s, 3H), 3.71 (s, 3H), 2.62 (k, J= 7.5 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

b) 3-((5-Ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-7-carboxylic acid A21

To a suspension of methyl 3-(5-ethyl-2-methoxyphenylsulfonamido) benzo[ d ]isoxazole-7-carboxylate A20 (200 mg, 0.5 mmol) in MeOH (10 mL) and THF (10 mL) was added 2 M aq. NaOH (1.28 mL) and the mixture was stirred at rt overnight. The solvent was removed under reduced pressure, the residue was diluted with water and the pH was adjusted to 2-3. The resulting precipitate was collected by filtration to afford the title compound (144 mg, 75%) as an off-white solid. LCMS-D: sp _temp 2.43 min; m/z 377.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 13.5 (s, 1H), 11.8 (s, 1H), 8.35-8.29 (m, 1H), 8.15-8.09 (m, 1H), 7.72 (d, J= 2.3 Hz, 1H), 7.53-7.44 (m, 2H), 7.12-7.05 (m, 1H), 3.71 (s, 3H), 2.62 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

c) 3-(5-Ethyl-2-methoxyphenylsulfonamido)benzo[d]isoxazole-7-carbonyl chloride A22

A mixture of 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-7-carboxylic acid A21 (30 mg, 0.08 mmol) and _SOCl2 (5 mL) was heated at 85°C under _N2 for 3 h and then concentrated under reduced pressure to give the title compound (31 mg, 100%), which was used directly in the next step without further purification.

d) 3-(5-Ethyl-2-methoxyphenylsulfonamido)-N, N-dimethylbenzo[d]isoxazole-7-carboxamide 185

To a solution of 3-(5-ethyl-2-methoxyphenylsulfonamido)benzo[ d ]isoxazole-7-carbonyl chloride A22 (31 mg, 0.08 mmol) in THF (1 mL) was added dropwise dimethylamine (40% solution in water, 2 mL) and the mixture was stirred at rt for 1 h. The mixture was concentrated under reduced pressure, the residue was diluted with water, adjusted to pH 2-3 and extracted with DCM (30 mL x 3). The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC to give the title compound (18 mg, 56%) as a white solid. LCMS-D: _sp 2.36 min; m/z 404.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.9 (s, 1H), 8.21-8.06 (m, 1H), 7.78-7.56 (m, 2H), 7.45 (s, 2H), 7.17-6.97 (m, 1H), 3.71 (s, 3H), 3.01 (s, 3H), 2.80 (s, 3H), 2.61 (m, 2H), 1.15 (m, 3H).

Example 186: 5-ethyl -N- (7-(hydroxymethyl)benzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 186

A mixture of 3-(5-ethyl-2-methoxyphenylsulfonamido)benzo[ d ]isoxazole-7-carboxylic acid A21 (30 mg, 0.08 mmol) and _BH3⋅THF (1 M solution in THF, 3 mL, 3 mmol) was stirred at rt under _N2 for 5 h. The reaction was quenched with water and most of the THF was removed under reduced pressure. The residue was adjusted to pH 2-3 and extracted with DCM (15 mL×3). The combined organic extracts were dried over anhydrous _{Na2SO4 ,} filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM/MeOH=30/1) to give the title compound (20 mg, 71%) as a light red solid. LCMS-D: _sp 2.35 min; m/z 363.1 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 11.7 (s, 1H), 7.96-7.89 (m, 1H), 7.70 (d, J= 2.3 Hz, 1H), 7.61-7.55 (m, 1H), 7.46 (dd, J= 8.5, 2.3 Hz, 1H), 7.34 (t, J= 7.6 Hz, 1H), 7.12-7.05 (m, 1H), 5.39 (t, J= 5.7 Hz, 1H), 4.70 (d, J= 5.4 Hz, 2H), 3.73 (s, 3H), 2.61 (k, J= 7.2 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

Example 187: 5-ethyl-2-methoxy -N- (7-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 187

To a suspension of N- (7-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 137 (150 mg, 0.36 mmol) in 1,4-dioxane (18 mL) and water (4.5 mL) were added _K2CO3 ( ₁₅₀ mg, 1.09 mmol), methylboronic acid (45 mg, 0.73 mmol) and Pd(dppf) _Cl2 (27 mg, 0.036 mmol) and the mixture was heated at 90 °C under _N2 for 4 h. The mixture was concentrated under reduced pressure and the residue was partitioned between EtOAc (50 mL), water (40 mL) and 1 M aq. HCl (15 mL). The layers were separated and the organic layer was washed with 0.5 M aq. HCl solution (40 mL×2), brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the title compound (20 mg, 16%) as a white solid. LCMS-D: sp _temp 2.25 min; m/z 347.1 [M+H] ⁺ , 369.1 [M+Na] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.7 (s, 1H), 7.89-7.82 (m, 1H), 7.70 (d, J= 2.3 Hz, 1H), 7.50-7.38 (m, 2H), 7.25 (t, J= 7.6 Hz, 1H), 7.12-7.04 (m, 1H), 3.73 (s, 3H), 2.60 (k, J= 7.6 Hz, 2H), 2.39 (s, 3H), 1.15 (t, J= 7.6 Hz, 3H).

Example 188: 5-ethyl -N- (7-ethylbenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 188

a) N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxy-N-(7-vinylbenzo[d]isoxazol-3-yl)benzenesulfonamide A23

To a suspension of N- (7-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A17 (200 mg, 0.36 mmol) in 1,4-dioxane (15 mL) and water (3 mL) were added 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (110 mg, 0.71 mmol), _K2CO3 (148 mg, 1.07 mmol) and Pd(dppf) _Cl2 (26 mg, 0.036 mmol), and the mixture was heated at 90 °C under _N2 for 4 _h . The mixture was concentrated under reduced pressure, the residue was diluted with EtOAc (30 mL) and washed with water (25 mL). × 3). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=2/1) to give the title compound (95 mg, 52%) as a white solid. LCMS-D: _sp 3.28 min; m/z 509.0 [M+H] ⁺ .

b) 5-Ethyl-2-methoxy-N-(7-vinylbenzo[d]isoxazol-3-yl)benzenesulfonamide A24

A mixture of N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxy -N- (7-vinylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide A23 (95 mg, 0.18 mmol) and TFA (4 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=2/1) to give the title compound (40 mg, 61%) as a white solid. LCMS-D: _sp 2.78 min; m/z 359.1 [M+H] ⁺

c) 5-Ethyl-N-(7-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide 188

To a solution of 5-ethyl-2-methoxy -N- (7-vinylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide A24 (35 mg, 0.098 mmol) in EtOAc (5 mL) was added 10% Pd/C (7 mg) and the mixture was stirred at rt under H ₂ atmosphere overnight. The mixture was filtered, the filtrate was concentrated and the residue was purified by silica gel chromatography (pet. ether/EtOAc=4/1) to give the title compound (30 mg, 85%) as a white solid. LCMS-D: _sp 2.83 min; m/z 361.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.7 (s, 1H), 7.90-7.83 (m, 1H), 7.70 (d, J= 2.3 Hz, 1H), 7.50-7.40 (m, 2H), 7.28 (t, J= 7.6 Hz, 1H), 7.12-7.05 (m, 1H), 3.73 (s, 3H), 2.80 (k, J= 7.5 Hz, 2H), 2.61 (k, J= 7.6 Hz, 2H), 1.22 (t, J= 7.5 Hz, 3H), 1.15 (t, J= 7.6 Hz, 3H).

Example 189: N- (7-cyclopropylbenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 189

a) N-(7-cyclopropylbenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A25

To a suspension of N- (7-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-diethylbenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A17 (200 mg, 0.36 mmol) in 1,4-dioxane (15 mL) and _H2O (3 mL) were added cyclopropylboronic acid (61 mg, 0.71 mmol), _K2CO3 (148 mg, 1.07 mmol) and Pd(dppf) _Cl2 (26 mg, 0.036 mmol), and the mixture was heated at 90 _° C under _N2 for 4 h. The mixture was concentrated under reduced pressure, the residue was diluted with EtOAc (30 mL) and washed with water (25 mL × 3). The organic layer was dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=3/1) to give the title compound (140 mg, 75%) as a white solid. LCMS-D: _sp 3.34 min; m/z 523.2 [M+H] ⁺ .

b) N-(7-Cyclopropylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 189

A mixture of N- (7-cyclopropylbenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A25 (140 mg, 0.27 mmol) and TFA (6 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=2/1) to give the title compound (80 mg, 81%) as a white solid. LCMS-D: _sp 2.86 min; m/z 373.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.7 (s, 1H), 7.81 (dd, J= 7.0, 2.1 Hz, 1H), 7.70 (d, J= 2.3 Hz, 1H), 7.50-7.43 (m, 1H), 7.27-7.19 (m, 2H), 7.12-7.06 (m, 1H), 3.73 (s, 3H), 2.61 (k, J= 7.6 Hz, 2H), 2.17-2.10 (m, 1H), 1.15 (t, J= 7.5 Hz, 3H), 1.04-0.97 (m, 2H), 0.90-0.84 (m, 2H).

Example 190: N- (7-cyclohexylbenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 190

a) N-(7-(cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A26

To a suspension of N- (7-bromobenzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-

methoxybenzenesulfonamide A17 (200 mg, 0.36 mmol) in 1,4-dioxane (15 mL) and water (3 mL) were added cyclohex-1-en _- 1-ylboronic acid (90 mg, 0.71 mmol), Pd(dppf) _Cl2 (26 mg, 0.036 mmol) and _K2CO3 (148 mg, 1.07 mmol) and the mixture was heated at 90 °C under _N2 for 4 h. The mixture was concentrated under reduced pressure, the residue was diluted with EtOAc (30 mL) and washed with water (25 mL × 3). The organic layer was dried over anhydrous _Na2SO4 , _filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=3/1) to give the title compound (150 mg, 75%) as a white solid, which was used directly in the next step.

b) N-(7-(Cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide A27

A mixture of N- (7-(cyclohex-1-en-1-yl)benzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A26 (150 mg, 0.26 mmol) and TFA (7 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=2/1) to give the title compound (65 mg, 60%) as a white solid. LCMS-D: _sp 3.64 min; m/z 413 [M+H] ⁺ .

c) N-(7-Cyclohexylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 190

A mixture of N- (7-(cyclohex-1-en-1-yl)benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide A27 (65 mg, 0.15 mmol) and 10% Pd/C (13 mg) in EtOAc (10 mL) was stirred at rt under H ₂ for 3 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to give the title compound (40 mg, 61%) as a white solid. LCMS-D: _sp 3.34 min; m/z 415.2 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.7 (s, 1H), 7.87-7.82 (m, 1H), 7.70 (d, J= 2.3 Hz, 1H), 7.49-7.41 (m, 2H), 7.28 (t, J= 7.6 Hz, 1H), 7.12-7.06 (m, 1H), 3.72 (s, 3H), 2.62 (k, J= 7.6 Hz, 2H), 1.85-1.75 (m, 4H), 1.75-1.67 (m, 1H), 1.61-1.48 (m, 2H), 1.44-1.20 (m, 4H), 1.15 (t, J= 7.6 Hz, 3H).

Example 191: 5-ethyl-2-methoxy -N- (7-(1-methyl-1 H -pyrazol-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 191

a) N-((2,4-Dimethoxybenzyl)oxy)-5-ethyl-2-methoxy-N-(7-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide A29

To a mixture of N- (7-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A17 (150 mg, 0.27 mmol) in 1,4-dioxane ( ₁₅ mL) and _{H2O (3 mL) were added K2CO3 (} 110 mg, 0.80 mmol), (1-methyl- 1H- pyrazol-4-yl)boronic acid (67 mg, 0.33 mmol) and Pd(dppf) _Cl2 (20 mg, 0.027 mmol) and the mixture was heated at 90 °C under _N2 overnight. The mixture was diluted with 0.5 M aq. HCl (30 mL) and most of the organic solvent was removed under reduced pressure. The remaining aqueous mixture was extracted with DCM (40 mL x 3), the combined organic extracts were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM/MeOH=100/1) to give the title compound (80 mg, 53%) as a light yellow solid, which was used directly in the next step.

b) 5-Ethyl-2-methoxy-N-(7-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide 191

A mixture of N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxy -N- (7-(1-methyl- 1H -pyrazol-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide A29 (80 mg, 0.14 mmol) and TFA (3 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM/MeOH=80/1) to give the title compound (50 mg, 86%) as a light yellow solid. LCMS-D: _sp 2.59 min; m/z 413.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 8.34 (s, 1H), 8.06 (s, 1H), 7.91-7.84 (m, 2H), 7.72 (d, J= 2.3 Hz, 1H), 7.50-7.43 (m, 1H), 7.41-7.32 (m, 1H), 7.13-7.06 (m, 1H), 3.90 (s, 3H), 3.74 (s, 3H), 2.62 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.5 Hz, 3H).

Example 192: 5-Ethyl-2-methoxy -N- (7-(pyrimidin-5-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 192

To a suspension of N- (7-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 137 (200 mg, 0.49 mmol) in toluene (16 mL), water (8 mL) and isopropanol (8 mL) were added pyrimidin-5-ylboronic acid (181 mg, 1.46 mmol), _{K3PO4 (} 518 mg, 1.95 mmol) and Pd(dppf) _Cl2 (36 mg, 0.049 mmol) and the mixture was heated at 90 °C under _N2 overnight. The pH of the mixture was adjusted to 5-6 and extracted with EtOAc (20 mL×3). The combined organic extracts were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM/MeOH=40/1) to give the title compound (18 mg, 9%) as a brown solid. LCMS-D: _sp 2.49 min; m/z 411.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.9 (s, 1H), 9.34-9.18 (m, 2H), 8.17 (d, J= 8.1 Hz, 1H), 8.06 (d, J= 7.4 Hz, 1H), 7.73 (s, 1H), 7.62-7.42 (m, 3H), 7.17-7.06 (m, 1H), 3.75 (s, 3H), 2.62 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

Example 193: 5-ethyl-2-methoxy -N- (6-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 193

a) N-(6-Bromobenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A30

To a solution of N- (6-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 138 (4.1 g, 10.0 mmol), (2,4-dimethoxyphenyl)methanol (2.5 g, 15.0 mmol) and _PPh3 (6.6 g, 25.0 mmol) in THF (100 mL) at 0 °C under _N2 was added DIAD (4.0 g, 20.0 mmol) and the mixture was stirred at rt overnight. The mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (pet. ether/EtOAc=50/1 to 5/1) to give the title compound (4.0 g, 71%) as a white solid, which was used directly in the next step.

b) N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxy-N-(6-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide A31

A mixture of N- (6-bromobenzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A30 (120 mg, 0.214 mmol), _CH3B (OH) ₂ (64 mg, 1.07 mmol), Pd(dppf) _Cl2 (31 mg, 0.428 mmol) and _K2CO3 (148 mg, 1.07 mmol) _in 1,4-dioxane (10 mL) and water (2 mL) was heated at 90°C under _N2 overnight. The solvent was removed under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 5/1) to give the title compound (72 mg, 68%) as a white solid, which was used directly in the next step.

c) 5-Ethyl-2-methoxy-N-(6-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide 193

A mixture of N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxy -N- (6-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide A31 (72 mg, 0.145 mmol) and TFA (3 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 3/1) to give the title compound (45 mg, 90%) as a white solid. LCMS-D: sp _temp 2.76 min; m/z 347.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 7.94-7.87 (m, 1H), 7.68 (d, J= 2.3 Hz, 1H), 7.49-7.42 (m, 1H), 7.39 (s, 1H), 7.23-7.16 (m, 1H), 7.13-7.05 (m, 1H), 3.72 (s, 3H), 2.60 (k, J= 7.5 Hz, 2H), 2.43 (s, 3H), 1.14 (t, J= 7.5 Hz, 3H).

Example 194: 5-ethyl -N- (6-ethylbenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 194

a) N-(2,4-Dimethoxybenzyl)-5-ethyl-N-(6-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide A32

A mixture of N- (6-bromobenzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A30 (200 mg, 0.356 mmol), ethyl boronic acid (132 mg, 1.78 mmol), Pd(dppf) _Cl2 (52 mg, 0.071 mmol), and _K2CO3 (246 mg, 1.78 mmol) in 1,4-dioxane (20 mL) and water (4 mL) was heated at 90 _° C under _N2 overnight. The solvent was removed under reduced pressure and the residue was partitioned between DCM (200 mL) and water (50 mL). The layers were separated, the organic layer was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 5/1) to give the title compound (120 mg, 67%) as a white solid, which was used directly in the next step.

b) 5-Ethyl-N-(6-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide 194

A mixture of N- (2,4-dimethoxybenzyl)-5-ethyl -N- (6-ethylbenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide A32 (120 mg, 0.24 mmol) and TFA (5 mL) was stirred at rt overnight and then concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 3/1) to give the title compound (80 mg, 94%) as a white solid. LCMS-D: _sp 2.84 min; m/z 361.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 7.97-7.90 (m, 1H), 7.69 (d, J= 2.3 Hz, 1H), 7.49-7.37 (m, 2H), 7.26-7.19 (m, 1H), 7.11-7.04 (m, 1H), 3.72 (s, 3H), 2.73 (k, J= 7.6 Hz, 2H), 2.60 (k, J= 7.6 Hz, 2H), 1.20 (t, J= 7.6 Hz, 3H), 1.14 (t, J= 7.6 Hz, 3H).

Example 195: N- (6-cyclopropylbenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 195

A mixture of N- (6-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 138 (206 mg, 0.5 mmol), cyclopropylboronic acid (215 mg, 2.5 mmol), Pd(dppf) _Cl2 (73 mg, 0.1 mmol) and _{K2CO3 (} 345 mg, 2.5 mmol) in 1,4-dioxane (20 mL) and water (4 mL) was heated at 90 °C under _N2 overnight. The solvent was removed under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 3/1) to give the title compound (80 mg, 43%) as a white solid. LCMS-D: sp _. 2.86 min; m/z 373.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 7.91-7.85 (m, 1H), 7.68 (d, J= 2.3 Hz, 1H), 7.48-7.42 (m, 1H), 7.27 (s, 1H), 7.12-7.04 (m, 2H), 3.72 (s, 3H), 2.60 (k, J= 7.6 Hz, 2H), 2.10-2.00 (m, 1H), 1.14 (t, J= 7.5 Hz, 3H), 1.07-0.99 (m, 2H), 0.83-0.75 (m, 2H).

Example 196: 5-ethyl-2-methoxy -N- (6-(1-methyl-1H-pyrazol-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 196

a) N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxy-N-(6-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide A33

A mixture of N- (6-bromobenzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A30 (120 mg, 0.214 mmol), (1-methyl- 1H- pyrazol-4-yl)boronic acid (54 mg, 0.428 mmol), Pd(dppf) _Cl2 (31 mg, 0.043 _mmol ), and _K2CO3 (148 mg, 1.07 mmol) in 1,4-dioxane (10 mL) and water (2 mL) was heated at 90 °C under _N2 overnight. The solvent was removed under reduced pressure and the residue was partitioned between DCM (150 mL) and water (50 mL). The layers were separated, the organic layer was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 5/1) to give the title compound (90 mg, 75%) as a brown solid. LCMS-D: _sp 3.05 min; m/z 563.0 [M+H] ⁺ .

b) 5-Ethyl-2-methoxy-N-(6-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide 196

A mixture of N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxy -N- (6-(1-methyl- 1H -pyrazol-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide A33 (90 mg, 0.16 mmol) and TFA (5 mL) was stirred at rt overnight and then concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=50/1 to 5/1) to give the title compound (30 mg, 46%) as a brown solid. LCMS-D: _sp 2.57 min; m/z 413.2 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 8.29 (s, 1H), 8.06-7.95 (m, 2H), 7.78 (s, 1H), 7.70 (d, J= 2.3 Hz, 1H), 7.64-7.56 (m, 1H), 7.50-7.42 (m, 1H), 7.13-7.05 (m, 1H), 3.87 (s, 3H), 3.71 (s, 3H), 2.61 (k, J= 7.6 Hz, 2H), 1.15 (t, J= 7.5 Hz, 3H).

Example 197: 5-Ethyl-2-methoxy -N- (6-(pyrimidin-5-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 197

A mixture of N- (6-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 138 (100 mg, 0.24 mmol), pyrimidin-5-ylboronic acid (45 mg, 0.36 mmol), Pd( _PPh3 ) ₄ (28 mg, 0.024 mmol) and _{K2CO3 (} 166 mg, 1.2 mmol) in toluene (8 mL), water (8 mL) and isopropanol (2 mL) was heated at 90 °C under _N2 for 4 h. 2 M aq. NaOH (15 mL) was added and the mixture was stirred at rt for 20 min. The mixture was washed with EtOAc (20 mL×2) and then adjusted to pH 2 with conc. HCl solution and extracted with DCM (50 mL x 2). The combined organic extracts were washed with water, brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (DCM/MeOH=30/1) to give the title compound (15 mg, 15%) as a white solid. LCMS-D: sp _temp 2.97 min; m/z 411.0 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 9.24 (s, 3H), 8.21-8.15 (m, 1H), 8.11 (s, 1H), 7.85-7.78 (m, 1H), 7.72 (d, J= 2.3 Hz, 1H), 7.49-7.42 (m, 1H), 7.13-7.05 (m, 1H), 3.73 (s, 3H), 2.61 (k, J= 7.6 Hz, 2H), 1.15 (t, J= 7.6 Hz, 3H).

Example 198: 5-ethyl-2-methoxy -N- (4-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 198

a) N-(4-Bromobenzo[d]isoxazol-3-yl)-N-((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A34

To a solution of N- (4-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 129 (2.1 g, 5.1 mmol), (2,4-dimethoxyphenyl)methanol (1.3 g, 7.7 mmol) and _PPh3 (3.35 g, 12.8 mmol) in THF (200 mL) at 0 °C under _N2 atmosphere was added DIAD (3.1 g, 15.3 mmol) and the mixture was stirred at 0 °C for 1 h and then allowed to warm to rt and stirred overnight. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=5/1) to give the title compound (1.2 g, 42%) as a white solid. LCMS-D: _sp 3.40 min; m/z 583.0 [M+H] ⁺ .

b) N((2,4-Dimethoxybenzyl)oxy)-5-ethyl-2-methoxy-N-(4-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide A35

A mixture of N- (4-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A34 (200 mg, 0.36 mmol), methyl boronic acid (43 mg, _0.71 mmol), _K2CO3 (148 mg, 1.07 mmol) and Pd(dppf) _Cl2 (26 mg, 0.036 mmol) in 1,4-dioxane (15 mL) and water (3 mL) was heated at 90 °C under _N2 for 4 h. The pH of the mixture was adjusted to 2-3 and most of the solvent was removed under reduced pressure. The residue was diluted with water (10 mL) and extracted with DCM (25 mL × 3). The combined organic layers were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EA=5/1) to give the title compound (125 mg, 71%) as a white solid, which was used directly in the next step.

c) 5-Ethyl-2-methoxy-N-(4-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide 198

A mixture of N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxy -N- (4-methyl benzo[ d ]isoxazol-3-yl)benzenesulfonamide A35 (125 mg, 0.26 mmol) and TFA (3 mL) was stirred at rt for 3 h, then diluted with DCM (100 mL), washed with water, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=3/1) to give the title compound (85 mg, 92%) as a white solid. LCMS-D: _sp 2.69 min m/z 347.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.6 (s, 1H), 7.58-7.56 (m, 1H), 7.52-7.46 (m, 3H), 7.18-7.14 (m, 2H), 3.73 (s, 3H), 2.62-2.56 (m, 5H), 1.14 (t, J= 7.6 Hz, 3H)

Example 199: 5-ethyl -N- (4-ethylbenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 199

a) N-(2,4-Dimethoxybenzyl)-5-ethyl-N-(4-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide A36

A mixture of N- (4-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A34 (200 mg, 0.36 mmol), ethyl boronic acid (53 mg, 0.71 _mmol ), _K2CO3 (148 mg, 1.07 mmol) and Pd(dppf) _Cl2 (26 mg, 0.036 mmol) in 1,4-dioxane (15 mL) and _H2O (3 mL) was heated at 90 °C under _N2 for 4 h. The pH of the mixture was adjusted to 2-3 and most of the solvent was removed under reduced pressure. The residue was diluted with water (15 mL) and extracted with DCM (20 mL × 3). The combined organic layers were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=5/1) to give the title compound (120 mg, 66%) as a white solid, which was used directly in the next step.

b) 5-Ethyl-N-(4-ethylbenzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide 199

A mixture of N- (2,4-dimethoxybenzyl)-5-ethyl -N- (4-ethylbenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide A36 (120 mg, 0.23 mmol) and TFA (3 mL) was stirred at room temperature under N ₂ for 3 h, then diluted with DCM (100 mL), washed with water, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=3/1) to give the title compound (75 mg, 89%) as a white solid. LCMS-D: _sp 2.80 min 361.1 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 10.5 (s, 1H), 7.60-7.49 (m, 4H), 7.22-7.18 (m, 2H), 3.81 (s, 3H), 8.09 (k, J= 7.6 Hz, 2H), 2.59 (k, J= 7.6 Hz, 2H), 1.25 (t, J= 7.6 Hz, 3H), 1.14 (t, J= 7.6 Hz, 3H).

Example 200: N- (4-cyclopropylbenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 200

a) N-(4-Cyclopropylbenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A37

A mixture of N- (4-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A34 (200 mg, 0.36 mmol), cyclopropylboronic acid (61 mg, 0.71 _mmol ), _K2CO3 (148 mg, 1.07 mmol) and Pd(dppf) _Cl2 (26 mg, 0.036 mmol) in 1,4-dioxane (15 mL) and _H2O (3 mL) was heated at 90 °C under _N2 for 4 h. The pH of the mixture was adjusted to 2-3 and most of the solvent was removed under reduced pressure. The residue was diluted with water (15 mL) and extracted with DCM (20 mL × 3). The combined organic extracts were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=5/1) to give the title compound (120 mg, 66%) as a yellow solid, which was used directly in the next step.

b) N-(4-Cyclopropylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 200

A mixture of N- (4-cyclopropylbenzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A37 (130 mg, 0.25 mmol) and TFA (3 mL) was stirred at rt under _N2 for 3 h and then concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=3/1) to give the title compound (85 mg, 92%) as a yellow solid. LCMS-D: _sp 2.81 min, 373.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.6 (s, 1H), 7.58-7.57 (d, J= 2.0 Hz, 1H), 7.53-7.42 (m, 3H), 7.18-7.16 (m, 1H), 6.86-6.84 (m, 1H), 3.74 (s, 3H), 2.72 (m, 1H), 2.60 (k, J= 7.6 Hz, 2H), 1.14 (t, J= 7.6 Hz, 3H), 1.02-0.98 (m, 2H), 0.82-0.78 (m, 2H).

Example 201: N- (4-cyclohexylbenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 201

a) N-(4-(Cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A38

A mixture of N- (4-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A34 (200 mg, 0.36 mmol), cyclohex-1- _{en-1-ylboronic acid (90 mg, 0.71 mmol), K2CO3 (} 148 mg, 1.07 mmol) and Pd(dppf) _Cl2 (26 mg, 0.036 mmol) in 1,4-dioxane (15 mL) and _H2O (3 mL) was heated at 90 °C under _N2 for 4 h. Most of the solvent was removed under reduced pressure, the residue was diluted with water (30 mL), adjusted to pH 1-2 and extracted with DCM (35 mL × 3). The combined organic extracts _were dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=100/1) to give the title compound (150 mg, 75%) as a white solid, which was used directly in the next step.

b) N-(4-(Cyclohex-1-en-1-yl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide A39

A mixture of N- (4-(cyclohex-1-en-1-yl)benzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A38 (150 mg, 0.27 mmol) and TFA (3 mL) was stirred at rt under _N2 for 3 h and then concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=2/1) to give the title compound (80 mg, 73%) as a white solid, which was used directly in the next step.

c) N-(4-Cyclohexylbenzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 201

A mixture of N- (4-(cyclohex-1-en-1-yl)benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide A39 (80 mg, 0.19 mmol) and 10% Pd/C (16 mg) in EtOAc (10 mL) was stirred at rt under H ₂ for 3 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to give the title compound (75 mg, 94%) as a white solid. LCMS-D: _sp 3.24 min, 415.2 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 10.5 (s, 1H), 7.58-7.48 (m, 4H), 7.26 (d, J= 7.2 Hz, 1H), 7.19 (d, J= 8.8 Hz, 1H), 3.80 (s, 3H), 3.62-3.56 (m, 1H), 2.59 (k, J= 7.6 Hz, 2H), 1.91-1.71 (m, 5H), 1.45-1.23 (m, 5H), 1.15 (t, J= 7.6 Hz, 3H).

Example 202: 5-ethyl-2-methoxy -N- (4-(1-methyl-1 H -pyrazol-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 202

a) N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxy-N-(4-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide A40

A mixture of N- (4-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A34 (200 mg, 0.36 mmol), (1-methyl- 1H -pyrazol-4-yl)boronic _acid (90 mg, 0.71 mmol), _K2CO3 (148 mg, 1.07 mmol) and Pd(dppf) _Cl2 (26 mg, 0.036 mmol) in 1,4-dioxane (15 mL) and _H2O (3 mL) was heated at 90 °C under _N2 for 4 h. Most of the solvent was removed under reduced pressure, the residue was diluted with water (30 mL), adjusted to pH 1-2 and extracted with DCM (35 mL× 3). The combined organic extracts were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=100/1) to give the title compound (105 mg, 53%) as a yellow solid, which was used directly in the next step.

b) 5-Ethyl-2-methoxy-N-(4-(1-methyl-1H-pyrazol-4-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide 202

A mixture of N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxy -N- (4-(1-methyl- 1H -pyrazol-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide A40 (105 mg, 0.19 mmol) and TFA (2 mL) was stirred at rt under _N2 for 3 h and then concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=80/1) to give the title compound (60 mg, 79%) as a yellow solid. LCMS-D: _sp 2.587 min, m/z 413.1 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 10.2 (s, 1H), 8.12 (s, 1H), 7.90 (s, 1H), 7.66-7.45 (m, 4H), 7.39 (d, J= 7.2 Hz, 1H), 7.13 (d, J= 8.4 Hz, 1H), 3.88 (s, 3H), 3.75 (s, 3H), 2.58 (k, J= 7.6 Hz, 2H), 1.15 (t, J= 7.6 Hz, 3H).

Example 203: 5-Ethyl-2-methoxy -N- (4-(pyrimidin-5-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 203

a) N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxy-N-(4-(pyrimidin-5-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide A41

A mixture of N- (4-bromobenzo[ d ]isoxazol-3-yl) -N- ((2,4-dimethoxybenzyl)oxy)-5-ethyl-2-methoxybenzenesulfonamide A34 (200 mg, 0.36 mmol), pyrimidin-5-ylboronic acid (43 mg, 0.71 mmol), _K2CO3 (148 mg, 1.07 mmol) and Pd(dppf) _Cl2 (26 mg, 0.036 mmol) in 1,4-dioxane (15 mL) and _H2O (3 mL) was heated at 90 _° C under _N2 for 4 h. Most of the solvent was removed under reduced pressure, the residue was diluted with water (30 mL), adjusted to pH 1-2 and extracted with DCM (35 mL × 3). The combined organic extracts _were dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=60/1) to give the title compound (95 mg, 47%) as a yellow solid, which was used directly in the next step.

b) 5-Ethyl-2-methoxy-N-(4-(pyrimidin-5-yl)benzo[d]isoxazol-3-yl)benzenesulfonamide 203

A mixture of N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxy -N- (4-(pyrimidin-5-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide A41 (95 mg, 0.17 mmol) and TFA (2 mL) was stirred at rt under _N2 for 3 h and then concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=40/1) to give the title compound (45 mg, 65%) as a red solid. LCMS-D: _sp 2.50 min, m/z 411.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.5 (s, 1H), 9.24 (s, 1H), 8.97 (s, 2H), 7.86-7.79 (m, 2H), 7.53-7.38 (m, 3H), 7.13-7.11 (m, 1H), 3.75 (s, 3H), 2.55 (t, J= 7.6 Hz, 2H), 1.11 (t, J= 7.6 Hz, 3H).

Example 204: N- (5-ethylbenzo[ d ]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonamide 204

A mixture of N- (5-bromobenzo[ d ]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonamide 113 (219 mg, 0.5 mmol), ethyl boronic acid (185 mg, 2.5 mmol), Pd(dppf) _Cl2 (73 mg, 0.1 mmol) and _K2CO3 (345 mg, 2.5 mmol) in 1,4-dioxane (20 mL) and water (20 mL) was heated at ₉₀ °C under _N2 overnight. The pH of the mixture was then adjusted to 1 and most of the solvent was removed under reduced pressure. The residue was diluted with water (50 mL) and extracted with DCM (100 mL). The organic extract was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=10/1 to 2/1) followed by preparative TLC (pet. ether/EtOAc=1/1) to give the title compound (52 mg, 27%) as a white solid. LCMS-D: _sp 3.06 min, m/z 387.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.4 (s, 1H), 7.83 (s, 1H), 7.54 (s, 1H), 7.51-7.43 (m, 2H), 6.84 (s, 1H), 3.71 (s, 3H), 2.77-2.65 (m, 6H), 1.75-1.66 (m, 4H), 1.24 (t, J= 7.6 Hz, 3H).

Example 205: N- (5-cyclopropylbenzo[ d ]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonamide 205

A mixture of N- (5-bromobenzo[ d ]isoxazol-3-yl)-3-methoxy-5,6,7,8-tetrahydronaphthalene-2-sulfonamide 113 (219 mg, 0.5 mmol), cyclopropylboronic acid ₍ 215 mg, 2.5 mmol), Pd(dppf) _Cl2 (73 mg, 0.1 mmol) and _K2CO3 (345 mg, 2.5 mmol) in 1,4-dioxane (20 mL) and water (4 mL) was heated at 90 °C under _N2 overnight. The pH of the mixture was then adjusted to 1 with conc. HCl and most of the solvent was removed under reduced pressure. The residue was diluted with water (30 mL) and extracted with DCM (100 mL). The organic extract was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=10/1 to 2/1) followed by preparative TLC (pet. ether/EtOAc=1/1) to give the title compound (50 mg, 25%) as a white solid. LCMS-D: _sp 3.07 min, m/z 399.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.4 (s, 1H), 7.68 (s, 1H), 7.54 (s, 1H), 7.47-7.43 (m, 1H), 7.40-7.35 (m, 1H), 6.84 (s, 1H), 3.71 (s, 3H), 2.77-2.65 (m, 4H), 2.08-2.00 (m, 1H), 1.74-1.66 (m, 4H), 1.04-0.96 (m, 2H), 0.68-0.61 (m, 2H).

Example 206: N- (4-chloro-5-ethylbenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 206

A mixture of N- (5-bromo-4-chlorobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 128 (200 mg, 0.449 mmol), ethyl boronic acid (166 mg, 2.244 mmol), Pd(dppf) _Cl2 (66 mg, 0.09 mmol), and _K2CO3 (310 mg, 2.244 mmol) in 1,4-dioxane (20 mL) and water (4 mL) was heated at 90 _° C under _N2 overnight. The pH of the mixture was then adjusted to 2 with 1 M aq. HCl and most of the solvent was removed under reduced pressure. The residue was diluted with water and extracted with DCM. The organic extract was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=50/1 to 2/1) followed by preparative TLC (DCM/MeOH=10/1) to give the title compound (15 mg, 9%) as a white solid. LCMS-D: _sp 2.89 min, m/z 395.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.4 (s, 1H), 7.64 (s, 2H), 7.61-7.57 (m, 1H), 7.49-7.44 (m, 1H), 7.15-7.10 (m, 1H), 3.66 (s, 3H), 2.80 (k, J= 7.6 Hz, 2H), 2.60 (k, J= 7.6 Hz, 2H), 1.21-1.11 (m, 6H).

Example 207: N- (4-chloro-5-cyclopropylbenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 207

A mixture of N- (5-bromo-4-chlorobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 128 (200 mg, 0.449 mmol), cyclopropylboronic acid (193 mg, 2.244 mmol), Pd(dppf) _Cl2 (66 mg, 0.09 mmol), and _K2CO3 (310 mg, 2.244 mmol) in 1,4-dioxane (20 mL) and water (4 mL) was heated at 90 _° C under _N2 overnight. The pH of the mixture was then adjusted to 2 with 1 M aq. HCl and most of the solvent was removed under reduced pressure. The residue was diluted with water and extracted with DCM. The organic extract was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=50/1 to 3/1) followed by preparative TLC (DCM/MeOH=100/1) to afford the title compound (20 mg, 11%) as a white solid. LCMS-D: _sp 2.91 min, m/z 407.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.4 (s, 1H), 7.64-7.56 (m, 2H), 7.52-7.44 (m, 1H), 7.34-7.27 (m, 1H), 7.17-7.11 (m, 1H), 3.67 (s, 3H), 2.60 (k, J= 7.6 Hz, 2H), 2.27-2.17 (m, 1H), 1.15 (t, J= 7.6 Hz, 3H), 1.08-1.00 (m, 2H), 0.78-0.70 (m, 2H).

Example 208: N- (5-cyclopropyl-4-methoxybenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 208

A mixture of N- (5-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 108 (100 mg, 0.227 mmol), cyclopropylboronic acid (97 mg, 1.133 mmol), Pd(dppf) _Cl2 (33 mg, 0.046 mmol), and _K2CO3 (156 mg, 1.133 mmol) _in 1,4-dioxane (20 mL) and water (4 mL) was heated at 90 °C under _N2 overnight. The pH of the mixture was then adjusted to 2 with 1 M aq. HCl and most of the solvent was removed under reduced pressure. The residue was diluted with water and extracted with DCM. The organic extract was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (pet. ether/EtOAc=50/1 to 3/1) followed by preparative TLC (DCM/MeOH=100/1) to afford the title compound (30 mg, 33%) as a white solid. LCMS-D: _sp 2.87 min; m/z 403.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 10.4 (s, 1H), 7.63 (d, J= 2.3 Hz, 1H), 7.50-7.45 (m, 1H), 7.32-7.26 (m, 1H), 7.19-7.12 (m, 2H), 3.92 (s, 3H), 3.81 (s, 3H), 2.61 (k, J= 7.6 Hz, 2H), 2.22-2.15 (m, 1H), 1.15 (t, J= 7.5 Hz, 3H), 1.03-0.94 (m, 2H), 0.76-0.64 (m, 2H).

Examples 209, 210 and 211

a) tert-Butyl((3,5-dimethoxybenzyl)oxy)dimethylsilane A42

To a solution of (3,5-dimethoxyphenyl)methanol (10 g, 0.059 mol) and imidazole (6.07 g, 0.089 mol) in DMF (100 mL) at rt was added TBS-Cl (9.86 g, 0.065 mol) and the mixture was stirred overnight. Water was added and the mixture was extracted with EtOAc. The organic extract was washed with water, concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=20/1 to 10/1) to give the title compound (13 g, 77%) as a colorless oil, which was used directly in the next step.

b) Lithium 4-(((tert-butyldimethylsilyl)oxy)methyl)-2,6-dimethoxybenzenesulfinate A43

To a solution of tert -butyl((3,5-dimethoxybenzyl)oxy)dimethylsilane A42 (5 g, 0.018 mol) and TMEDA (2.26 g, 0.020 mol) in n -hexane (100 mL) at -70 °C was added n- BuLi (2.5 M solution in hexanes, 7.8 mL, 0.020 mol) and the mixture was stirred at 0 °C for 1 h. The mixture was recooled to -60 °C and bubbled with SO ₂ gas for 30 min. The resulting precipitate was collected by filtration and washed with n -hexane to give the title compound (6.2 g, 100%), which was used directly in the next step.

c) 4-(((tert-Butyldimethylsilyl)oxy)methyl)-2,6-dimethoxybenzenesulfonyl chloride A44

To a mixture of lithium 4-((( tert -butyldimethylsilyl)oxy)methyl)-2,6-dimethoxy benzenesulfinate A43 (6.2 g, 0.018 mol) in n -hexane (100 ml) at 0°C was added _{SO2Cl2 (} 2.39 g, 0.018 mol), and the mixture was stirred at 0°C for 1 h. The solids were collected by filtration, washed with n -hexane, and then dissolved in EtOAc and washed with water. The organic layer was concentrated under reduced pressure, and the residue was purified by column chromatography (pet. ether/EtOAc=20/1 to 5/1) to give the title compound (2.4 g, 36%) as a yellow solid, which was used directly in the next step.

d) 4-(((tert-Butyldimethylsilyl)oxy)methyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide A45

To a solution of 4-methoxybenzo[ d ]isoxazol-3-amine (819 mg, 4.99 mmol) in THF (20 mL) at -78 °C was added LiHMDS (1 M solution in THF, 5.98 mL, 5.98 mmol) and the mixture was stirred at 0 °C for 30 min and then cooled to -60 °C. A solution of 4-((( tert -butyldimethylsilyl)oxy)methyl)-2,6-dimethoxybenzenesulfonyl chloride A44 (1.9 g, 4.99 mmol) in THF (20 mL) was added and the mixture was stirred at rt overnight. The pH of the mixture was adjusted to 5 with 1 M aq. HCl and extracted with EtOAc. The organic extract was concentrated under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 20/1) to give the title compound (170 mg, 7%) as a yellow solid. LCMS-D: _sp 3.33 min, m/z 509.0 [M+H] ⁺ .

e) 4-(Hydroxymethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide 209

To a solution of 4-((( tert -butyldimethylsilyl)oxy)methyl)-2,6-dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide A45 (170 mg, 0.334 mmol) in THF (10 mL) was added TBAF (175 mg, 0.668 mmol), and the mixture was stirred at rt for 3 h. The reaction was quenched with 1 M aq. HCl and the mixture was extracted with EtOAc. The organic extract was concentrated under reduced pressure, and the residue was purified by preparative TLC (DCM/MeOH=100/1 to 50/1) to give the title compound (30 mg, 23%) as a white solid. LCMS-D: temp. _spec. 1.93 min, m/z 394.9 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.40 (s, 1H), 7.55 (t, J= 8.2 Hz, 1H), 7.18-7.12 (m, 1H), 6.89-6.82 (m, 1H), 6.72 (s, 2H), 5.39 (t, J= 5.8 Hz, 1H), 4.49 (d, J= 5.8 Hz, 2H), 3.95 (s, 3H), 3.77 (s, 6H).

f) 4-(Bromomethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide A46

A solution of 4-(hydroxymethyl)-2,6-dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide 209 (1.0 g, 2.53 mmol), _CBr4 (2.52 g, 7.60 mmol) and _PPh3 (1.99 g, 7.60 mmol) in DCM (20 mL) was stirred at rt overnight. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography (DCM/MeOH=50/1 to 10:1) to give the title compound (600 mg, 52%) as a yellow solid, which was used directly in the next step.

g) 4-(Azidomethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide A47

A solution of 4-(bromomethyl)-2,6-dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide A46 (200 mg, 0.44 mmol) and _NaN3 (85 mg, 1.31 mmol) in DMF (5 mL) was heated at 50 °C for 3 h. The mixture was poured into water and the precipitate was collected by filtration. The filter cake was washed with water and dried to give the title compound (100 mg, 54%) as a gray solid. LCMS-D: _sp 2.45 min, m/z 420.1 [M+H] ⁺ .

h) 4-(Aminomethyl)-2,6-dimethoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide hydrochloride 210

A solution of 4-(azidomethyl)-2,6-dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide A47 (300 mg, 0.716 mmol) and _PPh3 (207 mg, 0.788 mmol) in THF (8 mL) and EtOH (6 mL) was stirred at rt for 3 h. Water (2 mL) and concentrated HCl (2 mL) were added and the mixture was heated at 55 °C for 3 h. Most of the organic solvents were removed under reduced pressure and the aqueous residue was washed with DCM. The aqueous phase was concentrated under reduced pressure to give the title compound (150 mg, 53%) as a yellow solid. LCMS-D: temp. _spec. 0.38 min, m/z 394.1 [M+H] ⁺ . ¹ H NMR (400 MHz, methanol- d ₄ ) δ 7.54 (t, J= 8.2 Hz, 1H), 7.06-7.01 (m, 1H), 6.87 (s, 2H), 6.86-6.82 (m, 1H), 4.12 (s, 2H), 4.07 (s, 3H), 3.89 (s, 6H).

i) N-(3,5-dimethoxy-4-(N-(4-methoxybenzo[d]isoxazol-3-yl)sulfamoyl)benzyl)acetamide 211

A mixture of 4-(aminomethyl)-2,6-dimethoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide hydrochloride 210 (130 mg, 0.33 mmol), _NaHCO3 (83 mg, 0.99 mmol) and acetic anhydride (101 mg, 0.99 mmol) in THF (5 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by preparative TLC to give the title compound (50 mg, 34%) as a yellow solid. LCMS-D: _sp 1.93 min, m/z 436.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.47 (s, 1H), 8.36 (t, J= 5.8 Hz, 1H), 7.55 (m, 1H), 7.18-7.12 (m, 1H), 6.89-6.81 (m, 1H), 6.66 (s, 2H), 4.24 (d, J= 5.9 Hz, 2H), 3.94 (s, 3H), 3.76 (s, 6H), 1.88 (s, 3H).

Example 212: 2-fluoro-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide 212

a) 2,6-Difluoro-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide A48

To a solution of 4-methoxybenzo[ d ]isoxazol-3-amine (4.0 g, 24.0 mmol) in THF (200 mL) at -78 °C under nitrogen atmosphere was added dropwise LiHMDS (1 M solution in THF, 31.2 mL, 31.2 mmol) and the mixture was stirred at -78 °C for 3 h. Then, a solution of 2,6-difluorophenylsulfonyl chloride (10.2 g, 48 mmol) in THF (20 mL) was added dropwise and the mixture was allowed to warm slowly to rt and stirred overnight. Water was added and the mixture was extracted with EtOAc (200 mL x 3). The combined organic extracts were washed with water, brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=6/1 to 2/1) to give the title compound (2.9 g, 35%) as a white solid. LCMS-D: _sp 1.94 min, m/z 341.1 [M+H] ⁺ .

b) 2-Fluoro-6-methoxy-N-(4-methoxybenzo[d]isoxazol-3-yl)benzenesulfonamide 212

A mixture of 2,6-difluoro -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide A48 (450 mg, 1.32 mmol) and NaOMe (428 mg, 7.92 mmol) in MeOH (6 mL) was heated at 120 °C overnight and then allowed to cool to rt and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative TLC (DCM/MeOH=50/1) to give the title compound (100 mg, 21%) as a white solid. LCMS-D: _sp 2.22 min, m/z 353.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.6 (s, 1H), 7.62 (m, 1H), 7.56 (t, J= 8.3 Hz, 1H), 7.18 (d, J= 8.4 Hz, 1H), 7.04 (d, J= 8.5 Hz, 1H), 7.00-6.92 (m, 1H), 6.84 (d, J= 8.0 Hz, 1H), 3.85 (s, 3H), 3.80 (s, 3H).

Example 213: 2-hydroxy-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide 213

A mixture of 2-fluoro-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide 212 (20 mg, 0.06 mmol), (6-methoxypyridin-3-yl)methanol (50.1 mg, 0.36 mmol) and t -BuOK (20.2 mg, 0.18 mmol) in NMP (1 mL) was heated at 120 °C for 16 h. LCMS analysis showed that only the hydrolysis product F was formed. The pH of the mixture was adjusted to < 4 with aq. HCl, diluted with water and extracted with EtOAc. The organic extract was washed with water, brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The procedure was repeated with another batch of 2-fluoro-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide 212 (52 mg, 0.15 mmol), the two crude products were combined and purified by prep. TLC (EtOAc/pet. ether = 1/13, then DCM/MeOH = 50/1 to 100/1) to give the title compound (135 mg, 19%) as a white solid. LCMS-D: _sp 2.39 min; m/z 350.9 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 10.2 (s, 1H), 7.57 (t, J= 8.4 Hz, 1H), 7.41 (t, J= 8.4 Hz, 1H), 7.16 (d, J= 8.4 Hz, 1H), 6.84 (d, J= 8.0 Hz, 1H), 6.60-6.56 (m, 2H), 3.91 (s, 3H), 3.75 (s, 3H).

Example 214: 2-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)-6-(methylamino)benzenesulfonamide 214

A solution of 2-fluoro-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide 212 (30 mg, 0.085 mmol), methylamine HCl (23 mg, 0.34 mmol) and _Et3N (52 mg, 0.51 mmol) in ethanol (3 mL) was heated at 120 °C in a sealed tube overnight. The mixture was concentrated under reduced pressure and the residue was purified by prep. TLC (DCM) to give the title compound (4 mg, 13%) as a yellow solid. LCMS-D: sp _temp 2.49 min; m/z 364.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.56 (t, J= 8.0 Hz, 1H), 7.30 (t, J= 8.0 Hz, 1H), 7.14 (d, J= 8.0 Hz, 1H), 6.84 (d, J= 8.0 Hz, 1H), 6.33 (d, J= 8.0 Hz, 1H), 6.24 (d, J= 8.0 Hz, 1H), 3.94 (s, 3H), 3.70 (s, 3H), 2.79 (s, 3H).

Example 215: 2-(cyclopropylmethoxy)-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide 215

To a solution of 2-fluoro-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide 212 (30 mg, 0.08 mmol) in THF (4 mL) was added t -BuOK (27 mg, 0.24 mmol) followed by cyclopropylmethanol (17 mg, 0.24 mmol) and the mixture was heated at 50 °C overnight. The mixture was diluted with water and extracted with EtOAc (20 mL x 3). The combined organic extracts were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified with prep. TLC (pet. ether/EtOAc=1/1) to give the title compound (4.4 mg, 13%) as a grey solid. LCMS-D: _sp 2.54 min, m/z 405.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.33 (s, 1H), 7.57 (t, J= 8.0 Hz, 1H), 7.48 (t, J= 8.0 Hz, 1H), 7.15 (d, J= 8.4 Hz, 1H), 6.85 (d, J= 8.0 Hz, 1H), 6.77-6.75 (m, 2H), 3.91 (s, 3H), 3.89 (d, J= 6.8 Hz, 2H), 3.77 (s, 3H), 0.86-0.83 (m, 1H), 0.43-0.24 (m, 4H).

Example 216: 2-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)-6-((2-methoxypyridin-4-yl)methoxy)benzenesulfonamide 216

To a solution of 2-fluoro-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide 212 (30 mg, 0.08 mmol) in NMP (3 mL) was added t -BuOK (48 mg, 0.43 mmol) followed by (2-methoxypyridin-4-yl)methanol (118 mg, 0.85 mmol), and the mixture was stirred at rt for 6 h. The mixture was diluted with water, adjusted to pH 5-6 with 1 M aq. HCl and extracted with EtOAc (20 mL×3). The combined organic extracts were washed with brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified with prep. TLC (DCM/MeOH=40/1) to give the title compound (5 mg, 12%) as a yellow solid. LCMS-D: _sp 2.43 min, m/z 472.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.65 (s, 1H), 8.07 (d, J= 5.2 Hz, 1H), 7.51-7.39 (m, 2H), 7.07-6.98 (m, 3H), 6.79-6.75 (m, 3H), 5.22 (s, 2H), 3.84 (s, 3H), 3.83 (s, 3H), 3.81 (s, 3H).

Examples 217-222 (Table G)

Table G. The following examples were synthesized according to the methods described for Example 215 or 216. Variations in conditions are noted in the table.

R-OH Name and structure Analytical data Method Note 217 Iso-propanol
2-Isopropoxy-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide JHMS-C: vr. _beat 2.18 min, m/z 393.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.13 (s, 1H), 7.58 (t, J= 8.0 Hz, 1H), 7.49 (t, J= 8.0 Hz, 1H), 7.17 (d, J= 8.0 Hz, 1H), 6.87 (d, J= 8.0 Hz, 1H), 6.79-6.75 (m, 2H), 4.75-4.65 (m, 1H), 3.95 (s, 3H), 3.80 (s, 3H), 1.17 (d, J= 6.0 Hz, 6H). As for example 216 120 °C, 16 h;
Fluoride/base/R-OH ratio 1:3:3 218 Ethanol
2-ethoxy-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide JHMS-C: vr. _beat 2.03 min, m/z 379.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.13 (s, 1H), 7.58 (t, J= 8.0 Hz, 1H), 7.50 (t, J= 8.0 Hz, 1H), 7.17 (d, J= 8.0 Hz, 1H), 6.86 (d, J= 8.0 Hz, 1H), 6.78 (d, J= 8.0 Hz, 2H), 4.01 (k, J= 7.2 Hz, 2H), 3.94 (s, 3H), 3.76 (s, 3H), 1.12 (t, J= 7.2 Hz, 3H). As for example 216 120 °C, 16 h;
Fluoride/base/R-OH ratio 1:3:3 219
2-(3-Methoxy-2-( N -(4-methoxybenzo[ d ]isoxazol-3-yl)sulfamoyl)phenoxy)acetamide LCMS-D: _sp 0.31 min, m/z 408.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.0 (s, 1H), 7.61-7.45 (m, 4H), 7.15 (d, J= 8.4 Hz, 1H), 7.83-6.74 (m, 3H), 4.51 (s, 2H), 3.83 (s, 3H), 3.77 (s, 3H). As for example 216 120 °C, 16 h;
Fluoride/base/R-OH ratio 1:6:10;
Eluent: DCM/MeOH
= 80/1 220
2-(2-Hydroxyethoxy)-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: vr. _beat 2.11 min, m/z 395.0 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.70 (s, 1H), 7.57-7.48 (m, 2H), 7.15 (d, J= 8.4 Hz, 1H), 6.84-6.80 (m, 3H), 5.10 (br s, 1H), 4.06 (t, J= 5.0 Hz, 2H), 3.90 (s, 3H), 3.80 (s, 3H), 3.63 (t, J= 5.0 Hz, 2H). As for example 216 k.t. during the night
then 80 °C, 16 h;
fluoride/base/R-OH ratio 1:6:10;
Eluent: DCM/MeOH
= 60/1 221
2-Methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)-6-((1-methyl-1H-pyrazol-4-yl)methoxy)benzenesulfonamide LCMS-D: _sp 2.25 min, m/z 445.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 7.62 (s, 1H), 7.54 (t, J= 8.4 Hz, 1H), 7.46 (t, J= 8.4 Hz, 1H), 7.40 (s, 1H), 7.12 (d, J= 8.4 Hz, 1H), 6.86 (d, J= 8.4 Hz, 1H), 6.81 (d, J= 8.0 Hz, 1H), 6.76 (d, J= 8.4 Hz, 1H), 5.04 (s, 2H), 3.86 (s, 3H), 3.75 (s, 3H), 3.73 (s, 3H). As for example 215 50 °C, 16 h;
Fluoride/base/R-OH ratio 1:3:3 222
2-(Benzyloxy)-6-methoxy -N- (4-methoxybenzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: _sp 2.67 min, m/z 441.1 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.51-7.43 (m, 4H), 7.36-7.30 (m, 3H), 7.05 (d, J= 8.4 Hz, 1H), 6.79-6.72 (m, 3H), 5.25 (s, 2H), 3.89 (s, 3H), 3.75 (s, 3H). As for example 215 50 °C, 16 h;
Fluoride/base/R-OH ratio 1:3:3

Example 223: 2,6-Dimethoxy -N- (4-nitrobenzo[ d ]isoxazol-3-yl)benzenesulfonamide 223

To a solution of 4-nitrobenzo[ d ]isoxazol-3-amine I10 (478 mg, 2.63 mmol) in THF (9 mL) at -78 °C under nitrogen atmosphere was added LiHMDS (1 M solution in THF, 5.30 mL, 5.30 mmol) and the mixture was stirred at -78 °C for 3 h. Then 2,6-dimethoxysulfonyl chloride I111 (1.25 g, 5.27 mmol) was added, the mixture was allowed to warm to rt and stirred overnight. The reaction was quenched with water and the mixture was extracted with EtOAc (80 mL × 3). The combined organic extracts were washed with water (100 mL × 3), brine (50 mL × 3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=1/0 to 50/1) to give the title compound (108 mg, 11%) as a yellow solid. LCMS-D: sp _temp 2.23 min; m/z 380.1 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.2 (s, 1H), 8.19 (d, J= 8.4 Hz, 2H), 7.92 (t, J= 8.4 Hz, 1H), 7.79 (d, J= 8.4 Hz, 1H), 6.79 (d, J= 8.4 Hz, 2H), 3.71 (s, 6H).

Example 224: N- (4-aminobenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 224

To a solution of 2,6-dimethoxy -N- (4-nitrobenzo[ d ]isoxazol-3-yl)benzenesulfonamide 223 (70.0 mg, 0.18 mmol) in EtOAc (30 mL) was added 10% Pd/C (40.0 mg), and the mixture was stirred at rt under hydrogen atmosphere overnight. The catalyst was removed by filtration, washed with EtOAc, and the filtrate was concentrated under reduced pressure. The procedure was repeated with an additional crop of 2,6-dimethoxy -N- (4-nitrobenzo[ d ]isoxazol-3-yl)benzenesulfonamide 223 (30 mg, 0.079 mmol), the crude products were combined, purified twice by prep. TLC (DCM/MeOH=50/1) and twice by prep. HPLC to afford the title compound (2.5 mg, 3%) as a white solid. LCMS-D: _sp 2.25 min; m/z 350.1 [M+H] ⁺ . ¹ H NMR (400 MHz, FDMSO- d ₆ ) δ 7.52 (t, J= 8.4 Hz, 1H), 7.25 (t, J= 8.0 Hz, 1H), 6.80 (d, J= 8.4 Hz, 2H), 6.66 (d, J= 8.0 Hz, 1H), 6.43 (d, J= 8.0 Hz, 1H), 3.78 (s, 6H).

Example 225: N- (5-((1 H -pyrazol-1-yl)methyl)benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 225

a) N-(5-Bromobenzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A49

To a solution of N- (5-bromobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 135 (8.2 g, 19.9 mmol), (2,4-dimethoxyphenyl)methanol (5.03 g, 29.9 mmol) and _PPh3 (13.1 g, 49.9 mmol) in THF (400 mL) at 0 °C under nitrogen atmosphere was slowly added DIAD (8.06 g, 39.9 mmol) dropwise and the mixture was stirred at rt overnight. The solvent was removed under reduced pressure and the residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 5/1) to give the title compound (4.7 g, 42%) as a white solid. LCMS-D: vr. _beat 3.39 min, m/z 583.1 [M+Na] ⁺

b) Methyl 3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylate A50

A mixture of N- (5-bromobenzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A49 (1.5 g, 2.67 mmol), Et ₃ N (10 mL) and Pd(dppf)Cl ₂ (732 mg, 1 mmol) in MeOH (150 mL) was heated at 120 °C under carbon monoxide (20 bar) for 7 h. The reaction was repeated using 1.7 g of N- (5-bromobenzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A49 and 1.5 g of N- (5-bromobenzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamido A49, the mixtures were combined and concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=100/1 to 3/1) to give the title compound (2.2 g, 49%) as a white solid. LCMS-D: _sp 3.17 min, m/z 563.1 [M+Na] ⁺

c) 3-((N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylic acid A51

To a solution of methyl 3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carboxylate A50 (2.2 g, 4.07 mmol) in methanol (30 mL) was added 3 M aq. NaOH (20 mL) and the mixture was stirred at rt overnight. The pH of the mixture was adjusted _to 1 with aq. HCl and the solvent was removed under reduced pressure. The residue was diluted with water (100 mL) and extracted with DCM (200 mL). The organic extract was washed with water, brine, dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure to give the title compound (1.9 g, 89%) as a light yellow solid. LCMS-D: vr. _beat 2.99 min, m/z 549.1 [M+Na] ⁺ .

d) N-(2,4-Dimethoxybenzyl)-5-ethyl-N-(5-(hydroxymethyl)benzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide A52

To a solution of 3-(( N -(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido) benzo[ d ]isoxazole-5-carboxylic acid (800 mg, 1.52 mmol) A51 in THF (5 mL) at 0 °C under nitrogen atmosphere was slowly added dropwise BH ₃ ⋅THF (1 M solution in THF, 10 mL, 10 mmol), the mixture was allowed to warm to rt and stirred for 2 h. The mixture was cooled to 0 °C and the reaction was quenched by slow addition of methanol (20 mL). The solvent was removed under reduced pressure, the residue was diluted with water and extracted with DCM (200 mL). The organic extract was washed with water, brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=100/0 to 40/1) to give the title compound (700 mg, 90%) as a white solid. LCMS-D: _sp 2.99 min, m/z 513.1 [M+H] ⁺ .

e) N-(5-((1H-Pyrazol-1-yl)methyl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A53

To a solution of N- (2,4-dimethoxybenzyl)-5-ethyl -N- (5-(hydroxymethyl)benzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide A52 (200 mg, 0.39 mmol) and _Et3N (197 mg, 1.95 mmol) in DCM (10 mL) at 0°C under nitrogen was added methanesulfonyl chloride (89 mg, 0.78 mmol) and the mixture was stirred at rt for 1.5 h and then used directly in the next step without workup and isolation.

To a solution of pyrazole (133 mg, 1.95 mmol) in DMF (20 mL) at 0 °C under nitrogen atmosphere was added t -BuOK (219 mg, 1.95 mmol) portionwise and the mixture was stirred at rt for 1.5 h. Then the reaction mixture containing (3-(( N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazol-5-yl)methyl methanesulfonate (ca. 0.39 mmol) was slowly added and the resulting mixture was stirred at rt for 30 min. The mixture was diluted with water and extracted with EtOAc. The organic extract was washed with water (x 3), brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=40/1) to give the title compound (60 mg, 27%) as a white solid. LCMS-D: _sp 3.01 min, m/z 563.2 [M+H] ⁺ .

f) N-(5-((1H-Pyrazol-1-yl)methyl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 225

A mixture of N- (5-((1 H -pyrazol-1-yl)methyl)benzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A53 (60 mg, 0.107 mmol) and TFA (5 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=20/1) to give the title compound (30 mg, 68%) as a white solid. LCMS-D: _sp 6.03 min, m/z 412.5 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.9 (s, 1H), 7.89 (s, 1H), 7.85 (d, J= 2.0 Hz, 1H), 7.68 (d, J= 2.0 Hz, 1H), 7.58-7.49 (m, 3H), 7.46 (dd, J= 8.4, 2.0 Hz, 1H), 7.08 (d, J= 8.8 Hz, 1H), 6.31 (t, J= 2.0 Hz, 1H), 5.54 (s, 2H), 3.68 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

Example 226: N- (5-(aminomethyl)benzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide trifluoroacetate 226

a) 3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxamide A54

A mixture of 3-(( N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carboxylic acid A51 (600 mg, 1.14 mmol) and thionyl chloride (5 mL) was heated at 85 °C for 1.5 h and then concentrated under reduced pressure. The residue was dissolved in DCM (3 mL) and added dropwise to conc. NH ₄ OH (10 mL) at 0 °C and the mixture was stirred at rt for 30 min. Most of the solvent was removed under reduced pressure and the residue was diluted with water (20 mL). The resulting precipitate was collected by filtration, washed twice with water and dried to give the title compound (400 mg, 67%) as a light yellow solid. LCMS-D: ep. _beat 2.86 min, m/z 526.2 [M+H] ⁺ .

b) N-(5-(Aminomethyl)benzo[d]isoxazol-3-yl)-N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A55

A mixture of 3-(( N-( 2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carboxamide A54 (400 mg, 0.76 mmol) and _BH3THF (1 M solution in THF, 20 mL) was heated at 75 °C under nitrogen atmosphere overnight. The mixture was then cooled to 0 °C and the reaction was quenched by slow addition of methanol (10 mL). The solvent was removed under reduced pressure, the residue was diluted with water and extracted with DCM (100 mL). The organic extract was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=100/1 to 10/1) to give the title compound (150 mg, 39%) as a light yellow solid, which was used directly in the next step.

c) N-(5-(Aminomethyl)benzo[d]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide trifluoroacetate 226

A mixture of N- (5-(aminomethyl)benzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A55 (50 mg, 0.098 mmol) and TFA (5 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=10/1) to give the title compound (30 mg, 86%) as a white solid. LCMS-D: _sp 1.93 min, m/z 362.1[M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (br s, 1H), 8.53 (br s, 3H), 8.11 (s, 1H), 7.83 (d, J= 8.4 Hz, 1H), 7.70-7.68 (m, 2H), 7.48 (d, J= 8.0 Hz, 1H), 7.12 (d, J= 8.8 Hz, 1H), 4.13 (s, 2H), 3.71 (s, 3H), 2.64 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H).

Example 227: methyl ((3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazol-5-yl)methyl) carbamate 227

a) Methyl ((3-((N-(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazol-5-yl)methyl) carbamate A56

To a solution of N- (5-(aminomethyl)benzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A55 (70 mg, 0.137 mmol) and _Et3N (69 mg, 0.685 mmol) in DCM (20 mL) at 0 °C was added dropwise methyl chloroformate (39 mg, 0.411 mmol) and the mixture was stirred at rt for 20 min. The solvent was removed under reduced pressure, the residue was diluted with water and extracted with DCM. The organic extract was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=100/0 to 40/1) to give the title compound (30 mg, 43%) as a white solid, which was used directly in the next step.

b) Methyl ((3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazol-5-yl)methyl) carbamate 227

A mixture of methyl ((3-(( N -(2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido) benzo[ d ]isoxazol-5-yl)methyl)carbamate A56 (30 mg, 0.0558 mmol) and TFA (3 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=40/1) to give the title compound (15 mg, 64%) as a white solid. LCMS-D: _sp 2.45 min, m/z 420.1 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 11.7 (s, 1H), 7.91 (s, 1H), 7.79 (t, J= 5.6 Hz, 1H), 7.69 (d, J= 2.0 Hz, 1H), 7.55-7.43 (m, 3H), 7.09 (d, J= 8.4 Hz, 1H), 4.29 (d, J= 6.0 Hz, 2H), 3.73 (s, 3H), 3.58 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

Example 228: N- ((3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazol-5-yl)methyl)acetamide 228

a) N-((3-((N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazol-5-yl)methyl)acetamide A57

To a solution of N- (5-(aminomethyl)benzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A55 (100 mg, 0.195 mmol) and _Et3N (98 mg, 0.975 mmol) in DCM (10 mL) at 0 °C under nitrogen atmosphere was added acetyl chloride (31 mg, 0.391 mmol) and the mixture was stirred at rt for 30 min. The mixture was diluted with water and extracted with DCM. The organic extract was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified with prep. TLC (DCM/MeOH=30/1) to give the title compound (60 mg, 56%) as a white solid, which was used directly in the next step.

b) N-((3-((5-Ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazol-5-yl)methyl)acetamide 228

A mixture of N- ((3-(( N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazol-5-yl)methyl)acetamide A57 (60 mg, 0.108 mmol) and TFA (5 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=40/1) to give the title compound (30 mg, 68%) as a white solid. LCMS-D: _sp 2.35 min, m/z 404.1 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 11.7 (br s, 1H), 8.55 (br s, 1H), 7.85 (s, 1H), 7.69 (s, 1H), 7.47-7.39 (m, 3H), 7.06 (d, J= 7.2 Hz, 1H), 4.33 (d, J= 3.6 Hz, 2H), 3.72 (s, 3H), 2.60 (k, J= 6.8 Hz, 2H), 1.91 (s, 3H), 1.17 (t, J= 6.8 Hz, 3H).

Example 229: 5-ethyl -N- (5-(hydroxymethyl)benzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide 229

a) Methyl 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylate A58

A mixture of methyl 3-(( N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxyphenyl)sulfonamido) benzo[ d ]isoxazole-5-carboxylate A50 (435 mg, 0.8 mmol) and TFA (9 mL) was stirred at rt for 3 h and then concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=2/1) to give the title compound (200 mg, 89%) as a light yellow solid. LCMS-D: _sp 2.69 min, m/z 391.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO -d6 ) δ 11.9 (br s, 1H), 8.82 (s, 1H), 8.18 (dd, J= 9.2, 1.6 Hz, 1H), 7.72-7.70 (m, 2H), 7.48 ( _dd , J= 8.8, 2.0 Hz, 1H), 7.10 (d, J= 8.8 Hz, 1H), 3.90 (s, 3H), 3.71 (s, 3H), 2.64 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

b) 3-((5-Ethyl-2-methoxyphenyl)sulfonamido)benzo[d]isoxazole-5-carboxylic acid A59

To a solution of methyl 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carboxylate A58 (220 mg, 0.56 mmol) in THF/MeOH (10 mL/10 mL) was added 2 M aq. NaOH (1.4 mL, 2.8 mmol) and the mixture was stirred at rt overnight. The solvent was removed under reduced pressure, the residue was dissolved in water (10 mL) and the pH of the solution was adjusted to 2-3. The resulting precipitate was collected by filtration to afford the title compound (180 mg, 85%) as a light yellow solid. LCMS-D: _sp 2.51 min, m/z 377.2 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 13.2 (s, 1H), 11.86 (s, 1H), 8.78 (s, 1H), 8.16 (dd, J= 8.8, 1.2 Hz, 1H), 7.71-7.66 (m, 2H), 7.47 (dd, J= 8.4, 1.6 Hz, 1H), 7.10 (d, J= 8.4 Hz, 1H), 3.70 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

c) 5-Ethyl-N-(5-(hydroxymethyl)benzo[d]isoxazol-3-yl)-2-methoxybenzenesulfonamide 229

A mixture of 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carboxylic acid A59 (50 mg, 0.13 mmol) and _BH3THF (1 M solution in THF, 5 mL, 5 mmol) was stirred at rt under nitrogen for 5 h. The reaction was quenched with water (10 mL) and most of the THF was removed under reduced pressure. The aqueous residue was adjusted to pH 2-3 and extracted with DCM (15 mL×3). The combined organic layers were dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM/MeOH=30/1) to give the title compound (30 mg, 62%) as a light yellow solid. LCMS-D: _sp 2.46 min, m/z 363.1 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 7.98 (s, 1H), 7.69 (d, J= 2.0 Hz, 1H), 7.53 (s, 2H), 7.46 (dd, J= 8.4, 2.0 Hz, 1H), 7.10 (d, J= 8.4 Hz, 1H), 5.35 (t, J= 5.6 Hz, 1H), 4.59 (d, J= 5.6 Hz, 2H), 3.72 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

Example 230: 3-((5-ethyl-2-methoxyphenyl)sulfonamido) -N- methylbenzo[ d ]isoxazole-5-carboxamide 230

A mixture of methyl 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carboxylate A58 (50 mg, 0.13 mmol) and methylamine (33% solution in EtOH, 5 mL) was heated in a sealed tube at 90 °C for 80 min. The solvent was removed under reduced pressure and the residue was purified by prep. TLC to give the desired product (18 mg, 36%) as a light yellow solid. LCMS-D: _sp 2.39 min, m/z 390.2[M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 8.53-8.51 (m, 2H), 8.03 (d, J= 8.8 Hz, 1H), 7.69 (d, J= 2.0 Hz, 1H), 7.62 (d, J= 8.8 Hz, 1H), 7.42 (d, J= 8.0 Hz, 1H), 7.06 (d, J= 8.4 Hz, 1H), 3.69 (s, 3H), 2.81 (d, J= 4.4 Hz, 3H), 2.62 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

Example 231: 3-((5-ethyl-2-methoxyphenyl)sulfonamido)- N, N -dimethylbenzo[ d ]isoxazole-5-carboxamide 231

A solution of 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carboxylic acid A59 (70 mg, 0.19 mmol) in thionyl chloride (10 mL) was heated at 85 °C for 3 h under nitrogen. The mixture was then concentrated under reduced pressure to give 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carbonyl chloride, which was dissolved in THF (2.5 mL) and treated with dimethylamine (40% solution in water, 5 mL). The mixture was stirred at rt overnight. Most of the THF was removed under reduced pressure and the pH of the aqueous residue was adjusted to 2-3. The precipitate was collected by filtration to afford the title compound (60 mg, 80%) as a light yellow solid. LCMS-D: _sp 2.40 min, m/z 404.2 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 8.13 (s, 1H), 7.71-7.63 (m, 3H), 7.48 (dd, J= 8.8, 2.0 Hz, 1H), 7.10 (d, J= 8.8 Hz, 1H), 3.70 (s, 3H), 3.01 (s, 3H), 2.96 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

Example 232: 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carboxamide 232

To a solution of 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carbonyl chloride (63 mg, 0.16 mmol), prepared according to the procedure described for 3-((5-ethyl-2-methoxyphenyl)sulfonamido)-N ,N- dimethylbenzo[ d ]isoxazole-5-carboxamide 231, in THF (2.5 mL) was added conc. NH ₄ OH (5 mL) and the mixture was stirred at rt over the weekend. Most of the THF was removed under reduced pressure and the aqueous residue was adjusted to pH 2-3. The resulting precipitate was collected by filtration to afford the title compound (37 mg, 62%) as a light yellow solid. LCMS-D: temp. _spec. 2.33 min, m/z 376.1 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 8.49 (s, 1H), 8.04 (d, J= 7.6 Hz, 2H), 7.69 (d, J= 2.4 Hz, 1H), 7.54 (d, J= 8.4 Hz, 2H), 7.36 (d, J= 6.8 Hz, 1H), 7.02 (d, J= 8.4 Hz, 1H), 3.68 (s, 3H), 2.61 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H).

Example 233: N- (5-cyanobenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide 233

A solution of 3-((5-ethyl-2-methoxyphenyl)sulfonamido)benzo[ d ]isoxazole-5-carboxamide 232 (22 mg, 0.058 mmol) in _POCl3 was heated at 110 °C overnight under nitrogen. The reaction was quenched with ice water (25 mL) and the mixture was extracted with DCM (35 mL x 2). The combined organic extracts were washed with water (50 mL x ₃ ), dried over anhydrous _Na2SO4 , filtered and concentrated under reduced pressure to give the title compound (19.8 mg, 90%) as a light yellow solid. LCMS-D: _sp 2.61 min, m/z 358.1 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 12.0 (s, 1H), 8.54 (s, 1H), 8.07 (d, J= 8.8, 1.2 Hz, 1H), 7.86 (d, J= 8.8 Hz, 1H), 7.73 (d, J= 2.0 Hz, 1H), 7.49 (dd, J= 8.4, 1.6 Hz, 1H), 7.11 (d, J= 8.4 Hz, 1H), 3.70 (s, 3H), 2.65 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H).

Example 234: 5-ethyl-2-methoxy -N- (5-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 234

a) N-(2,4-Dimethoxybenzyl)-5-ethyl-2-methoxy-N-(5-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide A60

A mixture of N- (5-bromobenzo[ d ]isoxazol-3-yl) -N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxybenzenesulfonamide A49 (112 mg, 0.2 mmol), _methyl boronic acid (60 mg, 1 mmol), Pd(dppf) _Cl2 (29 mg, 0.04 mmol) and _K2CO3 (138 mg, 1 mmol) in 1,4-dioxane (10 mL) and water (2 mL) was heated at 90 °C under nitrogen atmosphere overnight. The solvent was removed under reduced pressure, the residue was diluted with water (30 mL) and extracted with EtOAc (100 mL). The organic extract was washed with water, brine, dried over _{anhydrous Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep TLC (pet. ether/EtOAc=3/1) to give the title compound (60 mg, 61%) as a white solid. LCMS-D: _sp 2.72 min, m/z 347.2[M-DMB] ⁺ .

b) 5-Ethyl-2-methoxy-N-(5-methylbenzo[d]isoxazol-3-yl)benzenesulfonamide 234

A mixture of N- (2,4-dimethoxybenzyl)-5-ethyl-2-methoxy -N- (5-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide A60 (60 mg, 0.12 mmol) and TFA (5 mL) was stirred at rt for 4 h and then concentrated under reduced pressure. The residue was purified by prep. TLC (pet. ether/EtOAc=2/1) to give the title compound (30 mg, 71%) as a white solid. LCMS-D: _sp 2.72 min, m/z 347.2 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 7.78 (s, 1H), 7.69 (d, J= 2.0 Hz, 1H), 7.49-7.44 (m, 3H), 7.10 (d, J= 8.4 Hz, 1H), 3.72 (s, 3H), 2.61 (k, J= 7.6 Hz, 2H), 2.40 (s, 3H), 1.16 (t, J= 7.6 Hz, 3H).

Examples 235-238 (Table H)

Table H. The following target compounds were prepared according to the procedure described for 5-ethyl-2-methoxy -N- (5-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 234

Example Source materials Name and structure LCMS ¹ H NMR 235
5-Ethyl -N- (5-ethylbenzo[ d ]isoxazol-3-yl)-2-methoxybenzenesulfonamide LCMS-D: _sp 2.84 min, m/z 361.1 [M+H] ^{+ 1H} NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 7.83 (s, 1H), 7.69 (s, 1H), 7.51-7.44 (m, 3H), 7.10 (d, J= 8.4 Hz, 1H), 3.74 (s, 3H), 2.74 (k, J= 7.6 Hz, 2H), 2.63 (k, J= 7.6 Hz, 2H), 1.24 (t, J= 7.6 Hz, 3H), 1.16 (t, J= 7.6 Hz, 3H). 236
N -(5-Cyclopropylbenzo[ d ]isoxazol-3-yl)-5-ethyl-2-methoxybenzenesulfonamide LCMS-D: _sp 2.87 min, m/z 373.1 [M+H] ^{+ 1} H NMR (400 MHz, DMSO _-d6 ) δ 11.5 (s, 1H), 7.70 (d, J= 2.8 Hz, 1H), 7.69 (s, 1H), 7.47 (d, J= 8.8 Hz, 2H), 7.38-7.36 (m, 1H), 7.10 (d, J= 8.8 Hz, 1H), 3.74 (s, 3H), 2.63 (k, J= 7.6 Hz, 2H), 2.06-2.01 (m, 1H), 1.17 (t, J= 7.6 Hz, 3H), 1.02-0.96 (m, 2H), 0.68-0.64 (m, 2H). 237
5-Ethyl-2-methoxy -N- (5-(1-methyl-1 H -pyrazol-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: _sp 2.57 min, m/z 413.1 [M+H] ^{+ 1} H NMR (400 MHz, DMSO _-d6 ) δ 11.6 (s, 1H), 8.17 (s, 1H), 8.10 (s, 1H), 7.82-7.80 (m, 2H), 7.72 (d, J= 1.6 Hz, 1H), 7.60 (d, J= 8.8 Hz, 1H), 7.47 (dd, J= 8.4, 1.6 Hz, 1H), 7.11 (d, J= 8.8 Hz, 1H), 3.90 (s, 3H), 3.75 (s, 3H), 2.64 (k, J= 7.6 Hz, 2H), 1.16 (t, J= 7.6 Hz, 3H). 238
5-Ethyl-2-methoxy -N- (5-(pyrimidin-5-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: _sp 2.52 min, m/z 442.9 [M+H] ^{+ 1H} NMR (400 MHz, DMSO _-d6 ) δ 11.8 (s, 1H), 9.24 (s, 1H), 9.14 (s, 2H), 8.44 (s, 1H), 8.09 (d, J= 8.8 Hz, 1H), 7.80-7.74 (m, 2H), 7.48 (d, J= 7.6 Hz, 1H), 7.12 (d, J= 8.8 Hz, 1H), 3.74 (s, 3H), 2.65 (k, J= 7.6 Hz, 2H), 1.17 (t, J= 7.6 Hz, 3H).

Examples 239-242 (Table I)

Method IA:

To a solution of the amine (0.2 mmol, 1.0 equiv) in pyridine (2 mL) was added sulfonyl chloride (1.5 equiv) and the mixture was heated at 120 °C under microwave irradiation for 2 h. The mixture was partitioned between water and EtOAc, the layers were separated and the organic layer was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC to give the title compound. Variations of the above conditions are noted in Table I.

Table I. The following examples were synthesized according to the IA method. Variations in conditions are noted in the table.

Pr. Name and structure Analytical data Intermediate connections (if applicable) Method Notes 239
N- (7-Bromo-5-methylbenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide JHMS-C: vr. _beat 2.45 min; m/z 427.0, 429.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO _-d6 ) δ 11.5 (s, 1H), 7.85 (s, 1H), 7.73 (s, 1H), 7.48 (t, J =8.4 Hz, 1H), 6.75 (d, J =8.4 Hz, 2H), 3.74 (s, 6H), 2.40 (s, 3H). 2,6-Dimethoxy
benzenesulfone
yl chloride I111
7-Bromo-5-methylbenzo[ d ]isoxazol-3-amine I132 I.A. 0.2 equiv. DMAP was used.
The organic layer was washed with 0.1 M aq HCl during workup.
Rev. TLC (DCM/MeOH, 20/1) 240
2,6-Dimethoxy- N- (4-methoxy-6-((2,2,2-trifluoroethoxy)methyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide JHMS-C: vr. _beat 2.33 min; m/z 476.9 [M+H] ⁺ ; ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.59 (br s, 1H), 7.49 (s, 1H), 7.18-7.06 (m, 1H), 6.86-6.62 (m, 3H), 4.75 (s, 2H), 4.15-4.11 (m, 2H), 3.91 (s, 3H), 3.76 (s, 6H). 2,6-Dimethoxy
benzenesulfone
yl chloride I111
4-Methoxy-6-((2,2,2-trifluoroethoxy)methyl)benzo[ d ]isoxazol-3-amine I118 I.A. 1 ml of pyridine was used.
Prep. TLC (pet. ether/EtOAc=1/1) 241
2,6-Dimethoxy- N- (5-methyl-6-(pyridin-2-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide LCMS-D: _sp 3.32 min; m/z 426.0 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 11.4 (br s, 1H), 8.68 (d, J = 3.6 Hz, 1H), 7.96-7.90 (m, 2H), 7.59-7.43 (m, 4H), 6.76 (d, J = 8.4 Hz, 2H), 3.76 (s, 6H), 2.34 (s, 3H). 2,6-Dimethoxy
benzenesulfone
yl chloride I111
5-Methyl-6-(pyridin-2-yl)benzo[ d ]isoxazol-3-amine I127 I.A. 5 ml of pyridine were used.
0.1 equiv. DMAP was used.
Prep. TLC (pet. ether/EtOAc=1/1) 242
N- (6-(Difluoromethoxy)-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide LCMS-C temp _sp 2.13 min; m/z 430.9 [M+H] ⁺ ; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.73 (br s, 1H), 7.52 (t, J = 8.4 Hz, 1H), 7.40 (t, J = 73.2 Hz, 1H), 7.04 (d, J = 1.6 Hz, 1H), 6.78 (d, J = 8.4 Hz, 2H), 6.69 (d, J = 1.6 Hz, 1H), 3.91 (s, 3H), 3.76 (s, 6H). 2,6-Dimethoxybenzenesulfonyl chloride I111
6-(Difluoromethoxy)-4-methoxybenzo[ d ]isoxazol-3-amine I121 I.A. heated for 1 hour.
The pH of the aqueous phase was adjusted to pH 5 using 1 M aq. HCl solution during the workup.
Rev. TLC (DCM/MeOH=
50/1)

Example 243: N -(6-(3-cyanophenyl)-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 243

To a solution of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6 _- dimethoxybenzenesulfonamide 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) were added (3-cyanophenyl)boronic acid (34 mg, 0.226 mmol), _Na2CO3 (36 mg, 0.339 mmol) and Pd( _PPh3 ) ₄ (14 mg, 0.011 mmol), and the mixture was heated at reflux under _N2 overnight. The pH of the mixture was adjusted to 4-5 with 1 M aqueous HCl and extracted with EtOAc. The combined organic extracts were washed with water, brine, dried _{over Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=100/1) to give the title compound (24 mg, 46%) as a white solid. LCMS-C: _sp 2.44 min, m/z 466.0 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.70 (s, 1H), 8.34 (s, 1H), 8.16 (d, J =7.5 Hz, 1H), 7.90 (d, J =7.7 Hz, 1H), 7.70 (t, J =7.8 Hz, 1H), 7.57 (s, 1H), 7.51 (t, J = 8.4 Hz, 1H), 7.19 (s, 1H), 6.79 (d, J = 8.5 Hz, 2H), 4.03 (s, 3H), 3.78 (s, 6H).

Example 244: 2,6-dimethoxy- N -(4-methoxy-6-methylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 244

To a solution of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) were added methylboronic _acid (14 mg, 0.226 mmol), _Na2CO3 (36 mg, 0.339 mmol) and Pd( _PPh3 ) ₄ (14 mg, 0.011 mmol) and the mixture was heated at reflux under _N2 overnight. The pH of the mixture was adjusted to 4-5 with 1 M aqueous HCl and extracted with EtOAc. The combined organic extracts were washed with water, brine, dried _{over Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC to give the title compound (8 mg, 19%) as a white solid. LCMS-C: _sp 2.18 min, m/z 379.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 9.45 (s, 1H), 7.49 (t, J =8.5 Hz, 1H), 6.97 (s, 1H), 6.77 (d, J =8.5 Hz, 2H), 6.70 (s, 1H), 3.91 (s, 3H), 3.77 (s, 6H), 2.42 (s, 3H).

Example 245: 2,6-dimethoxy- N -(4-methoxy-6-(pyridin-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 245

To a solution of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) were added pyridin-4- _ylboronic acid (28 mg, 0.226 mmol), _Na2CO3 (36 mg, 0.339 mmol) and Pd( _PPh3 ) ₄ (14 mg, 0.011 mmol) and the mixture was heated at reflux under _N2 overnight. The pH of the mixture was adjusted to 4-5 with 1 M aqueous HCl and extracted with EtOAc. The combined organic extracts were washed with water, brine, dried _{over Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=30/1) to give the title compound (22 mg, 44%) as a white solid. LCMS-C: _sp 0.58 min, m/z 442.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 9.75 (s, 1H), 8.69 (d, J =5.3 Hz, 2H), 7.84 (d, J =5.3 Hz, 2H), 7.63 (s, 1H), 7.50 (t, J =8.4 Hz, 1H), 7.20 (s, 1H), 6.78 (d, J =8.5 Hz, 2H), 4.03 (s, 3H), 3.78 (s, 6H).

Example 246: 2,6-dimethoxy- N -(4-methoxy-6-(pyridin-3-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 246

To a solution of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6 _- dimethoxybenzenesulfonamide 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) were added 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (47 mg, 0.226 mmol), _Na2CO3 (36 mg, 0.339 mmol) and Pd( _PPh3 ) ₄ (14 mg, 0.011 mmol) and the mixture was heated at reflux under _N2 overnight. The pH of the mixture was adjusted to 4-5 with 1 M aqueous HCl solution and extracted with EtOAc. The combined organic extracts were washed with water, brine, dried _{over Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=30/1) to give the title compound (9 mg, 18%) as a white solid. LCMS-C: _sp 0.77 min, m/z 442.0 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.67 (s, 1H), 9.02 (d, J =2.5 Hz, 1H), 8.63 (dd, J =4.8, 1.6 Hz, 1H), 8.21 (dt, J =8.0, 2.0 Hz, 1H), 7.56 (s, 1H), 7.54-7.48 (m, 2H), 7.17 (s, 1H), 6.79 (d, J =8.5 Hz, 2H), 4.03 (s, 3H), 3.79 (s, 6H).

Example 247: 2,6-dimethoxy- N -(4-methoxy-6-(6-methoxypyridin-3-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 247

To a solution of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 89 (50 mg, 0.113 mmol) in 1,4-dioxane (8 mL) and water (2 mL) were added (6-methoxypyridin-3-yl) _boronic acid (35 mg, 0.226 mmol), _Na2CO3 (36 mg, 0.339 mmol) and Pd( _PPh3 ) ₄ (14 mg, 0.011 mmol) and the mixture was heated at reflux under _N2 overnight. The pH of the mixture was adjusted to 4-5 with 1 M aqueous HCl solution and extracted with EtOAc. The combined organic extracts were washed with water, brine, dried _{over Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=60/1) to give the title compound (45 mg, 85%) as a white solid. LCMS-C: _sp 2.33 min, m/z 472.0 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.61 (s, 1H), 8.63 (d, J =2.6 Hz, 1H), 8.15 (dd, J =8.7, 2.6 Hz, 1H), 7.51 (t, J =8.5 Hz, 1H), 7.47 (s, 1H), 7.10 (s, 1H), 6.94 (d, J =8.6 Hz, 1H), 6.79 (d, J =8.5 Hz, 2H), 4.02 (s, 3H), 3.91 (s, 3H), 3.79 (s, 6H).

Example 248: 2,6-dimethoxy- N -(4-methoxy-6-(3-methoxy-5-methylphenyl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 248

a) 2-(3-Methoxy-5-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane A61

A mixture of 1-bromo-3-methoxy-5-methylbenzene (500 mg, 2.49 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (1.9 g, 7.49 mmol), potassium acetate (977 mg, 9.96 mmol) and Pd(dppf) _Cl2 (196 mg, 0.25 mmol) in 1,4-dioxane (20 mL) was heated at reflux under _N2 for 3 h. The mixture was diluted with EtOAc (300 mL), washed with water (50 mL×3), and the organic layer was dried over _Na2SO4 , _filtered , concentrated under reduced pressure. The residue was purified by column chromatography (pet. ether/EtOAc=20/1) to give the title compound (300 mg, 49%) as a white solid. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.08 (s, 1H), 6.94 (d, J = 2.6 Hz, 1H), 6.87 (s, 1H), 3.73 (s, 3H), 2.30-2.25 (m, 3H), 1.28 (s, 12H).

b) 2,6-Dimethoxy-N-(4-methoxy-6-(3-methoxy-5-methylphenyl)benzo[d]isoxazol-3-yl)benzenesulfonamide 248

To a solution of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 89 (50 mg, 0.11 mmol) in 1,4-dioxane (8 mL) and water (2 mL) were added 2-(3-methoxy-5-methylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane A61 (55 _mg , 0.22 mmol), _K2CO3 (61 mg, 0.44 mmol) and Pd( _PPh3 ) ₄ (13 mg, 0.011 mmol) and the mixture was heated at reflux under _N2 overnight. The pH of the mixture was adjusted to 4-5 with 1 M aqueous HCl solution and extracted with EtOAc. The combined organic extracts were washed with water, brine, dried _{over Na2SO4} , filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC to give the title compound (4.3 mg, 8%) as a white solid. LCMS-C: _sp 2.56 min, m/z 485.3 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 9.62 (s, 1H), 7.51 (t, J =8.5 Hz, 1H), 7.43 (s, 1H), 7.18 (s, 1H), 7.10 (s, 1H), 7.05 (s, 1H), 6.83 (s, 1H), 6.79 (d, J =8.5 Hz, 2H), 4.02 (s, 3H), 3.81 (s, 3H), 3.79 (s, 6H), 2.36 (s, 3H).

Example 249: 2,6-dimethoxy- N -(4-methoxy-6-(2-methoxypyridin-4-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 249

To a solution of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6 _- dimethoxybenzenesulfonamide 89 (50 mg, 0.11 mmol) in 1,4-dioxane (4 mL) and water (1 mL) were added (2-methoxypyridin-4-yl)boronic acid (33 mg, 0.22 mmol), _Na2CO3 (35 mg, 0.33 mmol) and Pd( _PPh3 ) ₄ (13 mg, 0.011 mmol) and the mixture was heated at reflux under _N2 overnight. The mixture was poured into water and extracted with DCM. _The combined organic extracts were dried over _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified with prep. HPLC to afford the title compound (5 mg, 10%) as a white solid. LCMS-C: _sp 2.29 min, m/z 472.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO _-d6 ) δ 9.71 (br s, 1H), 8.26 (d, J = 5.4 Hz, 1H), 7.59 (s, 1H), 7.50 (t, J = 8.5 Hz, 1H), 7.42 (dd, J = 5.4, 1.6 Hz, 1H), 7.25 (s, 1H), 7.16 (s, 1H), 6.78 (d, J= 8.5 Hz, 2H), 4.02 (s, 3H), 3.90 (s, 3H), 3.78 (s, 6H).

Example 250: 2,6-dimethoxy- N -(4-methoxy-6-(5-methoxypyridin-3-yl)benzo[ d ]isoxazol-3-yl)benzenesulfonamide 250

To a solution of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 89 (50 mg, 0.11 mmol) in 1,4-dioxane (4 mL) and water (1 mL) were added 3-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine ( ₅₂ mg, 0.22 mmol), _Na2CO3 (35 mg, 0.33 mmol) and Pd( _PPh3 ) ₄ (13 mg, 0.011 mmol), and the mixture was heated at reflux under _N2 atmosphere overnight. The mixture was poured into water and extracted with DCM. The combined organic extracts were dried over _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC to afford the title compound (5 mg, 10%) as a white solid. LCMS-C: _sp 2.25 min, _m /z 472.0 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO _-d6 ) δ 9.67 (s, 1H), 8.63 (s, 1H), 8.38 (d, J =2.7 Hz, 1H), 7.81-7.79 (m, 1H), 7.59 (s, 1H), 7.51 (t, J =8.5 Hz, 1H), 7.18 (s, 1H), 6.79 (d, J= 8.5 Hz, 2H), 4.04 (s, 3H), 3.94 (s, 3H), 3.79 (s, 6H).

Example 251: N -(6-(3-cyano-5-methoxyphenyl)-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 251

a) 3-Methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile A62

A mixture of 3-bromo-5-methoxybenzonitrile (500 mg, 2.35 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (1.8 g, 7.07 mmol), potassium acetate (923 mg, 9.4 mmol) and Pd(dppf) _Cl2 ⋅DCM (196 mg, 0.24 mmol) was heated at reflux under _N2 for 4 h. Water was added and the mixture was extracted with EtOAc (20 mL × 3). The combined organic extracts were washed with brine, dried over _Na2SO4 , _filtered and concentrated under reduced pressure. The residue was purified with prep. TLC (EtOAc/pet. ether = 1/5) to give the title compound (600 mg, 98%) as a yellow oil. LCMS-C: _sp 2.66 min; m/z 260.0 [M+H] ⁺ .

b) N-(6-(3-Cyano-5-methoxyphenyl)-4-methoxybenzo[d]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 251

To a solution of N- (6-bromo-4-methoxybenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 89 (415 mg, 0.94 mmol) in 1,4-dioxane (80 mL) and water (20 mL) were added 3-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile _A62 (500 mg, 2.82 mmol), _Na2CO3 (399 mg, 3.77 mmol) and Pd( _PPh3 ) ₄ (116 mg, 0.1 mmol) and the mixture was heated at reflux under _N2 overnight. The mixture was diluted with water and extracted with EtOAc. The combined organic extracts _were dried over _Na2SO4 , filtered and concentrated under reduced pressure. The residue was purified by prep. TLC (DCM/MeOH=30/1) to give the title compound (40 mg, 8.6%) as a white solid. LCMS-C: _sp 2.48 min, m/z 495.9 [M+H] ⁺ . ^1H NMR (400 MHz _{, DMSO-d6} ) δ 9.69 (br s, 1H), 7.89 (s, 1H), 7.67 (s, 1H), 7.57 (m, 1H), 7.49 (s, 2H), 7.15 (s, 1H), 6.77 (d, J = 8.5 Hz, 2H), 4.02 (s, 3H), 3.90 (s, 3H), 3.77 (s, 6H).

Example 252: 2,6-dimethoxy- N -(5-methyl-7-phenylbenzo[ d ]isoxazol-3-yl)benzenesulfonamide 252

To a solution of N- (7-bromo-5-methylbenzo[ d ]isoxazol-3-yl)-2,6-dimethoxybenzenesulfonamide 239 (50 mg, 0.117 mmol) in 1,4-dioxane (8 mL) and water (2 mL) were added phenylboronic _acid (22 mg, 0.176 mmol), _Na2CO3 (50 mg, 0.468 mmol) and Pd( _PPh3 ) ₄ (14 mg, 0.012 mmol) and the mixture was heated at reflux under _N2 overnight. The mixture was diluted with water and extracted with EtOAc (30 mL×3). The combined organic extracts were washed with brine, dried over _Na2SO4 , _filtered and concentrated under reduced pressure. The residue was purified by prep TLC (pet. ether/EtOAc=3/1) to afford the title compound (40 mg, 80%) as a white solid. LCMS-C: _sp 2.49 min; m/z 425.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO -d ₆ ) δ 7.87-7.80 (m, 2H), 7.67-7.55 (m, 2H), 7.48 (t, J =7.7 Hz, 2H), 7.42-7.32 (m, 2H), 6.69 (d, J =8.4 Hz, 2H), 3.70 (s, 6H), 2.45 (s, 3H).

Tests

Protein preparation

KAT5

Molecular biology. A codon-optimized DNA sequence (for expression in Escherichia coli ) encoding amino acid residues 2–461 (Uniprot accession number Q92993-2) of the human KAT5 isoform was synthesized at GenScript USA Inc (Piscataway, NJ, USA). It was ligated into a modified E. coli expression vector pET43a designed to encode an N-terminal hexahistidine tag followed by a tobacco etch virus (TEV) protease cleavage site and the KAT5 sequence. The resulting protein sequence is shown below.

MGHHHHHHGTENLYFQGSAEVGEIIEGCRLPVLRRNQDNEDEWPLAEILSVKDISGRKLFYVHYIDFNKRLDEWVTHERLDLKKIQFPKKEAKTPTKNGLPGSRPGSPEREVKRKVEVVS PATPVPSETAPASVFPQNGAARRAVAAQPGRKRKSNCLGTDEDSQDSSDGIPSAPRMTGSLVSDRSHDDIVTRMKNIECIELGRHRLKPWYFSPYPQELTTLPVLYLCEFCLKYGRSLKC LQRHLTKCDLRHPPGNEIYRKGTISFFEIDGRKNKSYSQNLCLLAKCFLDHKTLYYDTDPFLFYVMTEYDCKGFHIVGYFSKEKESTEDYNVACILTLPPYQRRGYGKLLIEFSYELSKV EGKTGTPEKPLSDLGLLSYRSYWSQTILEILMGLKSESGERPQITINEISEITSIKKEDVISTLQYLNLINYYKGQYILTLSEDIVDGHERAMLKRLLRIDSKCLHFTPKDWSKRGKWAS*

Protein expression. To produce recombinant KAT5 protein, the expression plasmid was transformed into E. coli BL21 DE3 and grown with shaking at 37 °C in 1 L Terrific broth (TB) supplemented with 100 μg/mL ampicillin and 50 μM zinc until an OD600 of 0.8 was achieved. Cultures were transferred at 18 °C, protein expression was induced by adding isopropyl β-D-1-thiogalactopyranoside to a final concentration of 0.5 mM, and the cultures were shaken overnight for another 16 h. After expression, cell cultures were centrifuged at 5000 x g for 20 min and the cell pellet was stored frozen at -20 °C.

Protein purification. Protein purification was initiated by thawing the cell pellet (25 g wet weight) in lysis buffer (50 mM Hepes pH 7.4, 500 mM NaCl, 5 mM imidazole, 5% [v/v] glycerol, 0.1% [w/v] CHAPS, 2 mM 2-mercaptoethanol, 3 mM _MgCl2 , 0.5 mg/mL lysozyme, benzonase endonuclease [EMD Millipore], 1 mM PMSF, EDTA-free complete protease inhibitor tablets [Roche]) using a ratio of 6 mL buffer per g of cells. Cells were further lysed by sonication using a Misonix Liquid Processor (6 x 30 sec pulses, 60 [70 W] amplitude) and then centrifuged at 48,000 x g at 4 °C. The supernatant (cell lysate) was mixed with 20 ml Q-Sepharose FF resin (GE Healthcare) pre-equilibrated with Q buffer (20 mM Hepes pH 7.4, 1 M NaCl). The unbound Q-Sepharose FF fraction was then incubated with 5 ml cOmplete His-Tag Purification Resin (Roche) pre-equilibrated with IMAC wash buffer (20 mM Hepes pH 7.4, 500 mM NaCl, 35 mM imidazole). The resin was washed with IMAC wash buffer and bound KAT5 was eluted with IMAC elution buffer (20 mM Hepes, pH 7.4, 500 mM NaCl, 300 mM imidazole). The IMAC-eluted protein was immediately desalted in storage buffer (50 mM sodium citrate, pH 6.5, 500 mM NaCl, 5% [v/v] glycerol) using two HiPrep 26/10 desalting columns (GE Healthcare) in series. The desalted protein was further purified by passing through a HiLoad 26/60 Superdex 75 column pre-equilibrated in storage buffer. Finally, KAT5 protein was concentrated to 1.5 mg/mL using an Amicon Ultra centrifugal filtration device (Utra-15 MWCO 10 kDa), flash frozen in liquid nitrogen, and stored in a -70 °C freezer.

KAT6A

Molecular biology. The DNA sequence encoding amino acid residues 507-778 (Uniprot accession number Q92794-1) of human KAT6A was amplified by PCR and ligated into a modified E. coli pET expression vector designed to encode a NusA solubility tag followed by a hexahistidine tag, a tobacco etch virus (TEV) protease cleavage site, and the KAT6A sequence. The resulting protein sequence is shown below.

MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDVRVQIDRKSGDFDTFRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIE SVTFDRITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVILREDMLPRENFRPGDRVRGVLYSVRPEARGAQLFV TRSKPEMLIELFRIEVPEIGEEVIEIKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPAQFVINAMAPADVASI VVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMTVDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEI EGLDEPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLEDLAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDE ATSGSGHHHHHHSAGENLYFQGAMGRCPSVIEFGKYEIHTWYSSPYPQEYSRLPKLYLCEFCLKYMKSRTILQQHMKKCGWFHPPVNEIYRKNNISVFE VDGNVSTIYCQNLCLLAKLFLDHKTLYYDVEPFLFYVLTQNDVKGCHLVGYFSKEKHCQQKYNVSCIMILPQYQRKGYGRFLIDFSYLLSKREGQAGSP EKPLSDLGRLSYMAYWKSVILECLYHQNDKQISIKKLSKLTGICPQDITSTLHHLRMLDFRSDQFVIIRREKLIQDHMAKLQLNLRPVDVDPECLRWTP

Protein expression. To produce recombinant KAT6A protein, the expression plasmid was transformed into E. coli strain BL21 DE3 and grown with shaking at 37 °C in 1 L Terrific broth (TB) supplemented with 100 μg/mL ampicillin until an OD600 of 0.8 was achieved. Cultures were transferred at 18 °C, protein expression was induced by adding isopropyl β-D-1-thiogalactopyranoside to a final concentration of 0.5 mM, and the cultures were shaken overnight for another 16 h. After expression, cell cultures were centrifuged at 5000 x g for 20 min and the cell pellet was stored frozen at -20 °C.

Protein purification. Protein purification was initiated by thawing the cell pellet (40 g wet weight) in lysis buffer (25 mM Tris-HCl pH 7.8, 500 mM NaCl, 5 mM DTT, 0.01% [v/v] Triton-X 100, 5% [v/v] glycerol, 2 mM MgCl2, ₁₀ mM imidazole, 0.5 mg/mL lysozyme, benzonase endonuclease [EMD Millipore], 1 mM PMSF, EDTA-free complete protease inhibitor tablets [Roche]) using a ratio of 5 mL buffer per g of cells. Additional cell lysis was performed with 3 passes (15,000 psi) through an ice-cold Avestin C5 cell grinder and then centrifuged at 48,000 x g at 4°C. The supernatant (cell lysate) was filtered through a 5 μm filter and loaded onto a 5 ml HiTrap IMAC Sepharose FF column (GE Healthcare) pre-equilibrated with IMAC wash buffer (25 mM Tris-HCl pH 7.8, 500 mM NaCl, 5 mM DTT, 0.01% [v/v] Triton-X 100, 5% [v/v] glycerol, 20 mM imidazole) using a Profinia Affinity Chromatography Cleanup System (Bio-Rad). The IMAC column was then washed with IMAC wash buffer and the bound KAT6A protein was eluted with IMAC elution buffer (25 mM Tris-HCl pH 7.8, 500 mM NaCl, 5% [v/v] glycerol, 5 mM DTT, 250 mM imidazole). The IMAC buffer-eluted protein was further purified by passing through a HiLoad 26/60 Superdex 200 column pre-equilibrated in storage buffer (25 mM Tris-HCl pH 7.8, 500 mM NaCl, 5 mM DTT, 5% [v/v] glycerol). Finally, KAT6A protein was concentrated to ≤1 mg/mL using an Amicon Ultra centrifugal filtration device (Utra-15 MWCO 10 kDa), flash frozen in liquid nitrogen, and stored in a -70 °C freezer.

KAT6B was purchased from SignalChem, catalog ID: K315-381BG

KAT7

Molecular biology. A codon-optimized DNA sequence encoding amino acid residues 325–611 (Uniprot accession number O95251-1) of human KAT7 was synthesized at GenScript USA Inc (Piscataway, NJ, USA). It was ligated into a modified E. coli pET43a expression vector engineered to encode an N-terminal hexahistidine tag, followed by a tobacco etch virus (TEV) protease cleavage site and the KAT7 sequence. The resulting protein sequence is shown below.

MGHHHHHHGTENLYFQGSRLQGQITEGSNMIKTIAFGRYELDTWYHSPYPEEYARLGRLYMCEFCLKYMKSQTILRRHMAKCVWKHPPGDEIYRKGSISVFEVDGKKNKIYCQNLCLLAKLFLDHKTLYYDVEPFLFYVMTEADNTGCHLIGY FSKEKNSFLNYNVSCILTMPQYMRQGYGKMLIDFSYLLSKVEEKVGSPERPLSDLGLISYRSYWKEVLLRYLHNFQGKEISIKEISQETAVNPVDIVSTLQALQMLKYWKGKHLVLKRQDLIDEWIAKEAKRSNSNKTMDPSCLKWTPPKGTAS

Protein expression. To produce recombinant KAT7 protein, the expression plasmid was transformed into E. coli strain BL21 DE3 RIL and grown with shaking at 37 °C in 1 L Terrific broth (TB) supplemented with 100 μg/mL ampicillin and 50 μM zinc until an OD600 of 0.8 was achieved. Cultures were transferred at 18 °C, protein expression was induced by adding isopropyl β-D-1-thiogalactopyranoside to a final concentration of 0.5 mM, and the cultures were shaken overnight for another 16 h. After expression, cell cultures were centrifuged at 5000 x g for 20 min and the cell pellet was stored frozen at -20 °C.

Protein purification. Protein purification was initiated by thawing the cell pellet (10 g wet weight) in lysis buffer (50 mM Hepes, pH 7.5, 300 mM NaCl, 5 mM DTT, 5 mM imidazole, 0.05% [v/v] Brij 35, 10% [v/v] glycerol, 3 mM _MgCl2 , 0.5 mg/mL lysozyme, benzonase endonuclease [EMD Millipore], 1 mM PMSF, EDTA-free complete protease inhibitor tablets [Roche]) using a ratio of 10 mL buffer per g of cells. Cells were further lysed by sonication using a Misonix Liquid Processor (6 x 30 sec pulses, 60 [70 W] amplitude) and then centrifuged at 48,000 x g at 4 °C. The supernatant (cell lysate) was incubated with 1 ml cOmplete His-Tag Purification Resin (Roche) pre-equilibrated with IMAC Wash Buffer 1 (25 mM Hepes pH 7.5, 800 mM NaCl, 5 mM imidazole, 10% [v/v] glycerol, 5 mM DTT, 0.01% [v/v] Brij 35, 50 mM arginine, 50 mM glutamic acid). The resin was washed sequentially with IMAC wash buffer 1 and IMAC wash buffer 2 (25 mM Hepes pH 7.5, 300 mM NaCl, 20 mM imidazole, 10% [v/v] glycerol, 5 mM DTT, 0.01% [v/v] Brij 35, 50 mM arginine, 50 mM glutamic acid). Bound KAT7 protein was eluted with IMAC elution buffer (25 mM Hepes pH 7.5, 200 mM NaCl, 500 mM imidazole, 10% [v/v] glycerol, 5 mM DTT, 0.01% [v/v] Brij 35, 50 mM arginine, 50 mM glutamic acid). The eluted protein was collected directly in 4 volumes of desalting buffer (50 mM Na citrate, pH 6.5, 200 mM NaCl, 0.01% [v/v] Brij 35, 10% [v/v] glycerol, 5 mM DTT) to obtain a final imidazole concentration of 100 mM. The protein eluted by IMAC was immediately desalted in desalting buffer using two HiPrep 26/10 desalting columns (GE Healthcare) in series. The desalted protein was further purified by passing through a HiLoad 26/60 Superdex 75 column pre-equilibrated in storage buffer (50 mM Na citrate, pH 6.5, 200 mM NaCl, 10% [v/v] glycerol, 5 mM DTT). Finally, the KAT7 protein was concentrated to 3.5 mg/mL using an Amicon Ultra centrifugal filter device (Utra-15 MWCO 10 kDa), flash frozen in liquid nitrogen and stored in a -70 °C freezer.

KAT8

Molecular biology. A codon-optimized DNA sequence (for expression in E. coli) encoding amino acid residues 177–447 (Uniprot accession number Q9H7Z6-1) of human KAT8 was synthesized at Thermo Fisher Scientific GENEART GmbH (Regensberg, Germany). It was ligated into the E. coli expression vector pPROEX Hta, engineered to encode an N-terminal hexahistidine tag, and was followed by a tobacco etch virus (TEV) protease cleavage site and the KAT8 sequence. The resulting protein sequence is shown below.

MSYYHHHHHHDYDIPTTENLYFQGAKYVDKIHIGNYEIDAWYFSPFPEDYGKQPKLWLCEYCLKYMKYEKSYRFHLGQCQWRQPPGKEIYRKSNISVYEVDGKDHKIYCQNLCLLAKLFLDHKTLYFDVEPFVFYILTEVDRQGAHIV GYFSKEKESPDGNNVACILTLPPYQRRGYGKFLIAFSYELSKLESTVGSPEKPLSDLGKLSYRSYWSWVLLEILRDFRGTLSIKDLSQMTSITQNDIISTLQSLNMVKYWKGQHVICVTPKLVEEHLKSAQYKKPPITVDSVCLKWAP*

Protein expression. To produce recombinant KAT8 protein, the expression plasmid was transformed into E. coli BL21 DE3 and grown with shaking at 37 °C in 1 L Terrific broth (TB) supplemented with 100 μg/mL ampicillin until an OD600 of 0.8 was achieved. Cultures were transferred at 18 °C, protein expression was induced by adding isopropyl β-D-1-thiogalactopyranoside to a final concentration of 0.5 mM, and the cultures were shaken overnight for another 16 h. After expression, cell cultures were centrifuged at 5000 x g for 20 min and the cell pellet was stored frozen at -20 °C.

Protein purification. Protein purification was initiated by thawing the cell pellet (wet weight 34 g) in lysis buffer (20 mM Hepes pH 7.5, 500 mM NaCl, 5 mM imidazole, 5% [v/v] glycerol, 0.01% [v/v] Triton-X 100, 5 mM 2-mercaptoethanol, 2 mM _MgCl2 , 0.5 mg/ml lysozyme, benzonase endonuclease [EMD Millipore], 1 mM PMSF, complete protease inhibitor tablets without EDTA [Roche]) using a ratio of 3 ml buffer per 1 g of cells. Additional cell lysis was performed with 3 passes (15,000 psi) through an ice-cold Avestin C5 cell grinder and then centrifuged at 48,000 x g at 4°C. The supernatant (cell lysate) was filtered through a 0.2 μm filter and loaded onto a 5 ml HiTrap IMAC Sepharose FF column (GE Healthcare) pre-equilibrated with IMAC wash buffer 1 (20 mM Hepes, pH 7.5, 500 mM NaCl, 0.5 mM TCEP, 5 mM imidazole) using a Profinia Affinity Chromatography Purification System (Bio-Rad). The IMAC column was washed sequentially with IMAC wash buffer 1 and IMAC wash buffer 2 (20 mM Hepes pH 7.5, 500 mM NaCl, 0.5 mM TCEP, 10 mM imidazole) and the bound KAT8 protein was eluted with IMAC elution buffer (20 mM Hepes pH 7.5, 500 mM NaCl, 0.5 mM TCEP, 500 mM imidazole). The protein eluted with IMAC buffer was further purified by passing through a HiLoad 26/60 Superdex 200 column pre-equilibrated in storage buffer (20 mM Hepes pH 7.5, 500 mM NaCl, 1 mM TCEP). Finally, KAT8 protein was concentrated to ≤0.2 mg/mL using an Amicon Ultra centrifugal filtration device (Utra-15 MWCO 10 kDa), snap-frozen in liquid nitrogen, and stored in a -70 °C freezer.

Biochemical analysis of acetyltransferase

To determine the inhibition of KAT enzymatic activity by test compounds, assay reactions were performed in 8 μl volumes in 384-well low volume assay plates. Reactions were performed in assay buffer (100 mM Tris-HCl, pH 7.8, 15 mM NaCl, 1 mM EDTA, 0.01% Tween-20, 1 mM dithiothreitol, and 0.01% w/v hen egg white albumin).

Reactions were performed using 1 μM acetyl coenzyme A, 100 nM full-length recombinant histone tagged with limited biotinylation (KAT6A, KAT6B, KAT7: H3,1, KAT5, KAT8: H4), 10/ 5/ 8/ 40/ 20 nM KAT5/KAT6A/KAT6B/KAT7/KAT8 enzyme, respectively, and a specific antibody to acetyl lysine (H3,1: Cell Signaling Technology, H4: Abcam). Eleven point serial dilutions of test compounds were prepared in DMSO; a volume of 100 nL was transferred with a pin into the assay plates containing the substrates, after which the enzyme was added to initiate the reaction. Positive (no compound, DMSO only) and negative (no AcCoA) control reactions were included in the same plates and received the same amount of DMSO as the compound-treated wells. After addition of all reagents, the plates were sealed with plate adhesive films and incubated for 90 min at room temperature. An additional 4 μl of assay buffer containing lphaScreen® AA protein acceptor beads and streptavidin donor beads (PerkinElmer, Waltham, MA, USA) were then added to a final concentration of 8 μg/ml. After incubation for 2 h, the plate signal was measured using an EnVision 2103 Multilabel Plate Reader (PerkinElmer) in HTS AlphaScreen® mode. IC50 values were obtained from the raw data by calculating the percent inhibition (%I) for each reaction compared to controls on the same plate (%I=(I-CN)/(CP-CN), where CN/CP are the mean negative/positive reactions, respectively), then fitting the %I data versus compound concentration [I] to %I=(A+((BA)/(1+((C/[I])^D))), where A is the lower asymptote, B is the upper asymptote, C is the IC50 value, and D is the slope.

The results are presented in Tables 1-5 below:

Table 1 (TIP60-KAT5)

Example IC50 (µM) 1 > 125,000 2 > 125,000 3 = 65,106 4 = 35,221 5 = 114,325 6 = 94,934 7 > 125,000 8 > 125,000 9 = 40,976 10 = 93,664 11 > 125,000 12 > 125,000 13 = 119,896 14 > 125,000 15 = 7,294 16 = 30,179 17 = 27,659 18 = 118,055 19 = 64,983 20 = 81,458 21 > 125,000 22 = 38,877 23 = 72,865 24 > 125,000 25 = 120,445 26 > 125,000 27 > 125,000 28 > 125,000 29 > 125,000 30 > 125,000 31 > 125,000 32 = 56,003 33 = 90,452 34 > 125,000 35 = 33,836 36 = 38,979 37 > 125,000 38 = 80,086 39 > 125,000 40 = 121,024 41 = 65,079 42 = 11,568 43 = 23,002 44 = 60,208 45 = 34,341 46 > 125,000 47 = 123,081 48 = 92,895 49 = 74,577 50 > 125,000 51 = 87,660 52 > 125,000 53 > 125,000 54 > 125,000 55 > 125,000 56 > 125,000 57 > 125,000 58 > 125,000 59 > 125,000 60 > 125,000 61 = 85,401 62 > 125,000 63 > 125,000 64 > 125,000 65 > 125,000 66 = 123,371 67 = 114,876 68 > 125,000 69 = 124,236 70 = 36,766 71 = 28,431 72 > 125,000 73 = 2,161 74 = 8,132 75 = 33,535 76 = 2,578 77 = 51,770 78 > 125,000 79 = 17,451 80 = 10,913 81 = 31,488 82 = 8,247 83 = 17,898 84 = 50,464 85 = 28,466 86 = 97,635 88 > 125,000 89 = 5,785 90 = 90,684 91 = 2,183 92 = 1,387 93 = 53,063 94 = 124,242 95 = 13,967 96 = 28,640 97 = 6,181 98 = 31,286 99 = 9,699 100 = 51,711 101 = 98,204 102 = 9,330 103 = 69,326 104 = 28,112 105 > 125,000 106 = 14,254 107 = 74,084 108 = 41,946 109 = 69,262 110 > 125,000 111 = 26,608 112 = 16,494 113 = 4,836 114 = 59,355 115 = 23,660 116 = 10,463 117 > 125,000 118 = 37,082 119 = 8,726 120 = 5,781 121 = 29,010 122 = 58,591 123 = 59,355 124 = 27,879 125 > 125,000 126 = 51,346 127 = 49,699 128 = 6,301 129 = 18,536 130 > 125,000 131 = 69,740 132 = 53,030 133 = 45,717 134 = 51,523 135 = 4,788 136 > 125,000 137 = 14,209 138 = 5,115 139 = 34,292 140 = 47,841 141 > 125,000 142 = 32,183 143 > 125,000 144 = 11,487 145 = 20,967 146 = 21,951 147 > 125,000 148 > 125,000 149 > 125,000 150 = 50,012 151 > 125,000 152 = 38,945 153 = 4,055 154 = 25,313 155 = 0.672 156 > 125,000 157 > 125,000 158 > 125,000 159 > 125,000 160 = 46,217 161 > 125,000 162 = 13,954 163 > 125,000 164 > 125,000 165 > 125,000 166 = 3,093 167 = 32,157 168 = 3,415 169 = 63,197 170 = 4,826 171 = 4,374 172 = 9,070 173 = 4,050 174 = 1,833 175 = 43,215 176 = 5,670 177 = 7,082 178 = 7,535 179 = 32,129 180 = 25,476 181 = 23,827 182 = 65,635 183 = 33,231 184 > 125,000 185 > 125,000 186 = 84,702 187 = 65,279 188 = 69,912 189 = 31,983 190 = 19,884 191 = 68,651 192 = 29,888 193 = 53,125 194 = 37,844 195 = 16,708 196 = 95,399 197 > 125,000 198 = 73,654 199 = 14,892 200 = 4,733 201 = 21,848 202 = 110,730 203 = 74,251 204 = 4,565 205 = 3,729 206 = 6,361 207 = 4,605 208 = 28,461 209 = 28,674 210 > 125,000 211 = 7,931 212 = 9,302 213 > 125,000 214 > 125,000 215 > 125,000 216 = 123,671 217 > 125,000 218 > 125,000 219 > 125,000 220 > 125,000 221 > 125,000 222 = 26,758 223 > 125,000 224 > 125,000 225 > 125,000 226 = 80,348 227 > 125,000 228 = 124,363 229 > 125,000 230 > 125,000 231 > 125,000 232 > 125,000 233 > 125,000 234 = 10,877 235 = 9,044 236 = 10,055 237 > 125,000 238 > 125,000 239 = 17,373 240 = 122,168 241 = 26,350 242 = 27,314 243 = 18,076 244 = 30,172 245 = 57,554 246 = 27,066 247 = 96,592 248 = 9,331 249 = 69,187 250 = 12,220 251 = 46,983 252 = 101,708

Table 2 (MOZ-KAT6A)

Example IC50 (µM) 1 = 91,387 2 = 94,607 3 = 23,759 4 = 6,223 5 = 31,283 6 = 30,156 7 = 62,687 8 = 113,061 9 = 26,816 10 = 17,503 11 > 125,000 12 > 125,000 13 = 63,854 14 = 45,004 15 = 1,907 16 = 15,105 17 = 11,820 18 = 74,876 19 = 32,241 20 = 29,373 21 = 24,799 22 = 5,206 23 = 21,776 24 > 125,000 25 = 33,179 26 = 60,096 27 = 71,527 28 = 117,346 29 > 125,000 30 = 89,484 31 = 36,075 32 = 31,124 33 = 6,847 34 > 125,000 35 = 4,632 36 = 17,653 37 = 24,848 38 = 27,525 39 = 38,220 40 = 2,128 41 = 4,274 42 = 5,947 43 = 5,971 44 = 10,569 45 = 2,085 46 = 36,202 47 = 12,863 48 = 7,410 49 = 8,133 50 = 123,076 51 = 15,032 52 = 85,314 53 = 90,683 54 = 63,015 55 = 103,246 56 = 72,793 57 = 56,212 58 = 28,364 59 = 49,410 60 = 116,146 61 = 51,918 62 = 43,709 63 = 2,558 64 = 26,746 65 = 27,934 66 = 14,554 67 = 22,711 68 = 85,089 69 = 42,890 70 = 31,339 71 = 11,578 72 = 46,210 73 = 4,547 74 = 3,914 75 = 23,533 76 = 0.688 77 = 10,814 78 = 93,778 79 = 13,890 80 = 3,473 81 = 43,616 82 = 6,128 83 = 13,571 84 = 18,678 85 = 3,866 86 = 5,890 88 = 41,205 89 = 0.285 90 = 4,779 91 = 0,009 92 = 0.006 93 = 0.181 94 = 6,105 95 = 0,430 96 = 1,203 97 = 0.061 98 = 5,602 99 = 2,099 100 = 0.972 101 = 3,798 102 = 0.143 103 = 0.810 104 = 0.786 105 = 2,903 106 = 0.782 107 = 14,870 108 = 3,089 109 = 1,207 110 = 7,890 111 = 0.842 112 = 1,463 113 = 0.775 114 = 29,278 115 = 28,986 116 = 0,560 117 = 85,409 118 = 10,003 119 = 0,570 120 = 0.310 121 = 1,236 122 = 24,400 123 = 26,864 124 = 11,011 125 = 24,458 126 = 10,472 127 = 9,165 128 = 0,250 129 = 0.772 130 = 2,956 131 = 3,106 132 = 7,454 133 = 4,449 134 = 6,449 135 = 0.563 136 = 34,274 137 = 8,579 138 = 2,892 139 = 2,144 140 = 2,256 141 = 39,557 142 = 3,296 143 = 32,391 144 = 0.261 145 = 0.127 146 = 0.153 147 = 93,597 148 = 23,965 149 = 5,272 150 = 4,966 151 = 27,867 152 = 6,276 153 = 0.437 154 = 0.516 155 = 2,116 156 = 1,535 157 = 23,036 158 = 62,560 159 = 66,556 160 = 1,048 161 = 1,871 162 = 0.147 163 = 0.884 164 = 27,173 165 = 99,899 166 = 0.123 167 = 8,666 168 = 10,006 169 = 24,793 170 = 1,504 171 = 1,876 172 = 0.037 173 = 0,015 174 = 0,013 175 = 0.441 176 = 0,030 177 = 0.127 178 = 1,440 179 = 2,864 180 = 9,437 181 = 1,058 182 = 15,217 183 = 5,601 184 > 125,000 185 = 104,789 186 = 25,161 187 = 5,664 188 = 9,228 189 = 4,347 190 = 9,733 191 = 12,582 192 = 9,394 193 = 1,950 194 = 1,507 195 = 1,322 196 = 13,919 197 = 20,970 198 = 2,818 199 = 0.709 200 = 0.364 201 = 1,482 202 = 18,907 203 = 22,648 204 = 0,400 205 = 0.115 206 = 0.302 207 = 0.104 208 = 1,629 209 = 2,029 210 = 8,532 211 = 2,128 212 = 2,117 213 = 8,280 214 = 36,431 215 = 4,469 216 = 0.625 217 = 10,237 218 = 6,594 219 = 33,313 220 = 3,497 221 = 37,464 222 = 0.655 223 = 25,496 224 = 50,368 225 = 3,625 226 = 10,774 227 = 41,520 228 = 75,246 229 = 85,020 230 = 48,075 231 = 58,983 232 = 46,464 233 = 9,950 234 = 0.381 235 = 0.395 236 = 0.318 237 = 4,950 238 = 12,039 239 = 2,132 240 = 1,828 241 = 0.157 242 = 3,232 243 = 0.654 244 = 2,126 245 = 3,901 246 = 0.676 247 = 3,476 248 = 0.139 249 = 1,271 250 = 0.423 251 = 1,156 252 = 4,160

Table 3 (HBO-KAT7)

Example IC50 (µM) 1 = 53,948 2 = 15,521 3 = 23,243 4 = 5,168 5 = 6,011 6 = 6,277 7 = 14,175 8 > 125,000 9 = 8,418 10 = 54,053 11 = 60,488 12 = 49,922 13 = 63,834 14 = 15,174 15 = 1,456 16 = 9,635 17 = 14,575 18 = 34,064 19 = 36,094 20 = 41,258 21 = 25,506 22 = 6,300 23 = 23,893 24 = 33,854 25 = 41,948 26 = 34,465 27 = 32,121 28 = 54,786 29 = 94,098 30 = 11,481 31 = 53,590 32 = 14,923 33 = 11,409 34 = 31,493 35 = 8,748 36 = 26,267 37 = 114,461 38 = 6,698 39 = 10,116 40 = 19,929 41 = 11,845 42 = 8,384 43 = 12,914 44 = 10,794 45 = 6,833 46 = 74,439 47 = 29,419 48 = 54,767 49 = 77,831 50 > 125,000 51 = 28,277 52 > 125,000 53 > 125,000 54 > 125,000 55 > 125,000 56 > 125,000 57 > 125,000 58 = 16,607 59 > 125,000 60 > 125,000 61 > 125,000 62 = 70,618 63 = 60,060 64 = 105,707 65 = 59,720 66 = 113,991 67 > 125,000 68 > 125,000 69 = 94,149 70 = 32,029 71 = 21,593 72 = 11,413 73 = 0.508 74 = 1,665 75 = 5,748 76 = 0.937 77 = 12,022 78 > 125,000 79 = 4,059 80 = 1,129 81 = 6,726 82 = 1,496 83 = 3,792 84 = 20,038 85 = 1,769 86 = 1,981 88 = 7,509 89 = 0.168 90 = 18,889 91 = 0,079 92 = 0,060 93 = 2,799 94 = 16,059 95 = 0.754 96 = 4,265 97 = 0.444 98 = 3,411 99 = 5,739 100 = 2,240 101 = 32,782 102 = 0.568 103 = 5,928 104 = 3,132 105 = 63,160 106 = 3,977 107 = 11,732 108 = 2,038 109 = 4,067 110 = 17,497 111 = 4,536 112 = 3,014 113 = 0.914 114 = 41,609 115 = 95,520 116 = 1,435 117 > 125,000 118 = 0.852 119 = 1,146 120 = 0.503 121 = 5,211 122 = 4,122 123 = 4,198 124 = 13,017 125 = 85,834 126 = 28,884 127 = 9,487 128 = 0.618 129 = 1,318 130 = 21,712 131 = 23,780 132 = 11,785 133 = 5,342 134 = 27,644 135 = 1,426 136 = 28,316 137 = 6,558 138 = 3,683 139 = 10,043 140 = 9,895 141 = 24,336 142 = 1,165 143 = 7,883 144 = 0.286 145 = 0.259 146 = 0.511 147 > 125,000 148 = 36,783 149 = 9,432 150 = 8,039 151 = 25,571 152 = 9,866 153 = 0.507 154 = 3,128 155 = 0.248 156 = 1,975 157 = 15,514 158 = 44,862 159 = 37,620 160 = 1,577 161 = 2,506 162 = 0.969 163 = 13,103 164 = 61,638 165 = 30,654 166 = 0.328 167 = 6,854 168 = 36,401 169 = 8,646 170 = 0.982 171 = 0.292 172 = 0.426 173 = 0.169 174 = 0.031 175 = 5,949 176 = 0.138 177 = 0,021 178 = 0.482 179 = 13,481 180 = 1,759 181 = 5,137 182 = 15,608 183 = 4,480 184 = 30,983 185 = 60,714 186 = 47,092 187 = 21,961 188 = 40,080 189 = 8,016 190 = 18,459 191 = 1,576 192 = 24,230 193 = 15,518 194 = 6,872 195 = 4,963 196 = 23,280 197 = 45,687 198 = 6,951 199 = 0.632 200 = 0.678 201 = 0.773 202 = 54,375 203 = 23,360 204 = 3,045 205 = 0.936 206 = 0.626 207 = 0.257 208 = 4,654 209 = 1,054 210 = 6,539 211 = 0.917 212 = 1,048 213 = 20,519 214 > 125,000 215 = 16,630 216 = 3,106 217 = 18,406 218 = 6,444 219 = 60,688 220 = 11,008 221 = 63,721 222 = 1,422 223 = 43,526 224 > 125,000 225 = 31,135 226 = 37,985 227 = 63,139 228 > 125,000 229 = 115,218 230 = 109,417 231 > 125,000 232 > 125,000 233 = 17,872 234 = 3,739 235 = 0.820 236 = 1,070 237 = 18,016 238 = 94,180 239 = 3,003 240 = 10,198 241 = 2,510 242 = 1,785 243 = 0,720 244 = 1,113 245 = 2,761 246 = 0.478 247 = 5,897 248 = 0.375 249 = 0.889 250 = 0,050 251 = 0.473 252 = 1,303

Table 4 (MOF-KAT8)

Example IC50 (µM) 11 = 35,565 18 = 63,474 19 = 40,985 20 = 61,440 21 = 39,655 26 = 29,480 73 = 3,459 81 = 9,425 93 = 96,545 94 = 60,943 95 = 6,447 96 = 53,767 100 = 62,604 102 = 11,485 107 = 42,081 109 = 9,122 111 = 3,221 132 = 15,285 151 = 34,717 154 = 2,570 161 = 117,753 162 = 10,649 163 > 125,000 164 > 125,000 165 > 125,000 166 = 2,920 168 = 6,458 170 = 9,666 173 = 3,522 178 = 0,770 204 = 3,063 205 = 3,441 211 = 29,532 214 > 125,000 216 = 11,172 219 = 69,922 221 = 27,895 223 = 42,389 224 = 112,662 236 = 3,682

Table 5 (QKF-KAT6B)

Example IC50 (µM) 73 = 4,047 91 = 0.037 92 = 0.053 95 = 1,640 96 = 3,395 97 = 0.049 102 = 0.282 116 = 1,616 143 = 50,697 144 = 0.057 146 = 0.291 147 > 125,000 162 = 0,180 166 = 0.227 172 = 0.042 173 = 0,040 174 = 0,018 175 = 0.821 176 = 0.121 177 = 0.092 178 = 2,455 207 = 1,454

Analysis of the histone H3 lysine 23 acetylation biomarker

Compounds were tested for their ability to inhibit acetylation of the histone marker H3K23 in the following assay:

The U2OS cell line was seeded at a density of 9000 cells/well in 96-well optical tissue culture grade plates in RPMI medium and 10% fetal bovine serum and allowed to adhere for 24 hours under standard culture conditions (37 degrees Celsius, 5% CO2). At the end of this period, the medium was aspirated. Dilutions of compounds prepared in DMSO were added to the medium, with negative control wells treated with DMSO only, and positive control wells with 100% inhibition were treated with a strong inhibitor compound (e.g. cas 2055397-28-7, benzoic acid, 3-fluoro-5-(2-pyridinyl)-, 2-[(2-fluorophenyl)sulfonyl]hydrazide) (Baell, J., Nguyen, H.N., Leaver, D.J., Cleary, B.L., Lagiakos, H.R., Sheikh, B.N., Thomas. T.J., Aryl sulfonohydrazides, WO2016198507A1, 2016) at a concentration of 10 μM and 200 μl were transferred to the cells. After incubation for 24 hours, cells were fixed with 3.7% formaldehyde in phosphate-buffered saline (PBS) for 20 minutes at room temperature, washed (5 x 5 minutes) with phosphate-buffered saline containing 0.1% Tween 20, and blocked with Odyssey blocking buffer (LI-COR, Lincoln, NE, USA) containing 0.1% TritonX100. Specific antibody to H3K23ac (Abcam ab177275) in Odyssey blocking buffer containing 0.1% Tween 20 was added and incubated for 16 hours at 4 °C. After washing (as above), secondary antibody labeled with Alexa 647 (Life Technologies) and Hoechst 33342 (1 μg/ml, Sigma Aldrich) was added and incubated for 1 h. Plates were washed as previously described and read on a PerkinElmer Phenix simultaneous multiparameter imaging platform. Using a Columbus image analyzer pipeline, individual nuclei stained with Hoechst 33342 were localized and the level of acetylation was calculated from the intensity in the same region bound by Alexa647. The resulting mean intensity per cell was converted directly to percent inhibition relative to the control in the same plate and the data were fitted using a four-parameter logistic model to determine the 50% inhibition concentration (IC50).

The results are presented in Table 6 below.

Table 6

Example IC50 (µM) 73 > 10,000 86 = 1,579 91 = 0.182 92 = 0,013 95 > 10,000 96 > 10,000 97 = 0.045 116 = 3,455 144 = 0.128 145 = 0.098 146 = 0.045 147 > 10,000 155 > 10,000 162 = 0.144 166 = 0.398 172 = 0,014 173 = 0,020 174 = 0,017 176 = 0.685 177 = 1,486 178 > 10,000 191 > 10,000 242 = 0.672 250 = 3,713

Analysis of the histone H3 lysine 14 acetylation biomarker

Compounds were tested for their ability to inhibit acetylation of the histone H3 marker lysine 14 in the following assay:

The U2OS cell line was seeded at a density of 3,000 cells/well in 384-well optical-grade tissue culture plates in RPMI medium supplemented with 10% fetal bovine serum and 10 mM Hepes. Cells were allowed to adhere for 24 h under standard culture conditions (37 °C, 5% CO2). At the end of this period, cells were washed with serum-free medium. Dilutions of compounds prepared in DMSO were added to the serum-free medium, with negative control wells treated with DMSO only and positive control wells with 100% inhibition receiving a strong inhibitory compound (e.g., ( Z )-4-fluoro- N -((3-hydroxyphenyl)sulfonyl)-5-methyl-[1,1'-biphenyl]-3-carbohydrazonic acid) at a concentration of 10 μM. After incubation for 24 hours, cells were fixed with 4% formaldehyde in PBS for 15 minutes at room temperature, washed with phosphate-buffered saline containing 0.2% TritonX100 and 2% BSA. Specific antibody to H3K14ac (Cell Signaling Technologies) in blocking buffer was added and incubated overnight at 4 °C. After washing, secondary antibody labeled with AlexaFluor 488 (ThermoFisher) and Hoechst 33342 (1 μg/ml, Life Technologies) was added and incubated for 2 hours at room temperature. Plates were washed and read on a PerkinElmer Opera HCS simultaneous multiparameter imaging platform. Using the Columbus image analyzer pipeline, individual nuclei stained with Hoechst 33342 were localized and the level of acetylation was calculated from the intensity in the same region bound to AlexaFluor 488. The resulting mean intensity per cell was converted to percent inhibition relative to control in the same plate and the data were fitted using a four-parameter logistic model to determine the 50% inhibition concentration (IC50).

The results are presented in Table 7 below.

Table 7

Example IC50 (µM) 73 = 12,451 74 = 31,675 75 > 40,000 80 = 11,511 82 = 12,600 89 = 3,957 91 = 0.874 92 = 0.703 95 = 9,089 97 = 1,093 102 = 3,686 144 = 2,954 145 = 3,383 146 = 2,304 147 > 40,000 155 = 26,130 162 = 1,527 163 = 20,356 166 = 3,686 171 = 2,574 172 = 1,453 173 = 0.533 174 = 0.068 176 = 1,071 177 = 0.135 178 = 3,068 191 = 17,835 211 > 40,000 212 = 10,347 218 > 40,000 236 > 40,000 250 = 3,482 252 = 24,719

H2A.Z Lysine 7 Acetylation Biomarker Analysis

Compounds were tested for their ability to inhibit acetylation of the histone marker H2A.Z lysine 7 in the following assay:

The U2OS cell line was seeded at a density of 3,000 cells/well in 384-well optical tissue culture grade plates in RPMI medium supplemented with 10% fetal bovine serum and 10 mM Hepes. Cells were allowed to adhere for 24 hours under standard culture conditions (37 degrees Celsius, 5% CO2). At the end of this period, cells were washed with serum-free medium. Dilutions of compounds prepared in DMSO were added to the serum-free medium, with negative control wells treated with DMSO only and positive control wells with 100% inhibition receiving enantiomer 1 of the potent inhibitor compound 7-iodo- N- (2-(oxazol-2-yl)-2-phenylethyl)-2H - benzo[ e ][1,2,4]thiadiazine-3-carboxamide-1,1-dioxide, which is compound 146 from co-pending patent application GB1713962.7, filed August 31, 2018, at a concentration of 30 μM. After incubation for 24 hours, cells were fixed with 4% formaldehyde in PBS for 15 minutes at room temperature, washed with phosphate-buffered saline containing 0.2% TritonX100 and 2% BSA. Specific antibody to H2A.ZK7ac (Abcam) in blocking buffer was added and incubated overnight at 4 °C. After washing, secondary antibody labeled with AlexaFluor 488 (ThermoFisher) and Hoechst 33342 (1 μg/ml, Life Technologies) was added and incubated for 2 hours at room temperature. Plates were washed and read on a PerkinElmer Opera HCS simultaneous multiparameter imaging platform. Using the Columbus image analyzer pipeline, individual nuclei stained with Hoechst 33342 were localized and the level of acetylation was calculated from the intensity in the same region bound to AlexaFluor 488. The resulting mean intensity per cell was converted to percent inhibition relative to control in the same plate and the data were fitted using a four-parameter logistic model to determine the 50% inhibition concentration (IC50).

The results are presented in Table 8 below.

Table 8

Example IC50 (µM) 73 = 36,725 89 = 19,816 91 = 8,022 92 = 4,619 95 > 40,000 97 = 27,988 102 > 40,000 144 = 18,752 145 > 40,000 146 = 15,065 147 > 40,000 155 > 40,000 162 = 29,540 166 = 33,304 168 > 40,000 172 = 29,159 173 = 26,225 174 = 1,544 176 = 7,612 177 = 1,088 178 = 5,589 191 > 40,000 234 > 40,000 236 > 40,000 250 = 13,197 252 > 40,000

It is stated:

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof for use in a method of therapy:

(I)

where R ¹ , R ² , R ³ and R ⁴ are independently selected from:

(i) H;

(ii) C _1-3 alkyl, optionally substituted by:

hydroxy,

C _1-2 alkoxy, optionally substituted with one or more fluoro groups,

_NH2 ,

phenyl,

C _5-6 heteroaryl,

C _1-4 alkylcarbamoyl,

acylamido or

one or more fluoro groups;

(iii) C _1-3 alkoxy optionally substituted with C _3-6 cycloalkyl or one or more fluoro groups;

(iv) C _3-6 cycloalkyl;

(v) halogen;

(vi) COR ^C , where R ^C is selected from NR ^N1 R ^N2 , where R ^N1 and R ^N2 are independently selected from H and methyl;

(vii) cyano, NH ₂ or NO ₂ ; and

(viii) phenyl or C _5-6 heteroaryl optionally substituted with methyl, cyano, hydroxy or methoxy;

Ar is a phenyl, naphthyl or C _5-10 heteroaryl group, which groups are optionally substituted with one or more groups selected from:

(i) C _1-4 alkyl optionally substituted with hydroxy, C _1-2 alkoxy, NH ₂ , C _1-4 alkylcarbamoyl or one or more fluoro groups;

(ii) C _3-6 cycloalkyl;

(iii) hydroxy; cyano; NR ^N3 R ^N4 , where R ^N3 and R ^N4 are independently selected from H and methyl; or acylamido;

(iv) halogen;

(v) C _1-3 alkoxy optionally substituted with hydroxy, C(O)NH ₂ , C _3-6 cycloalkyl, phenyl, C _5-6 heteroaryl or one or more fluoro groups;

(vi) phenoxy optionally substituted with fluorine;

(vii) phenyl or C _5-6 heteroaryl;

(viii) SF ₅ or SO ₂ CH ₃ ;

(ix) -(CH ₂ ) _n -Y-, where Y is O or CH ₂ and n is 2 or 3; or

(x) C _1-4 alkyl ester.

2. A compound for use according to claim 1, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is H.

3. A compound for use according to any one of claims 1 or 2, characterized in that one of R ¹ , R ² , R ³ and R ⁴ is H.

4. A compound for use according to any one of claims 1 or 2, characterized in that two of R ¹ , R ² , R ³ and R ⁴ are H.

5. A compound for use according to any one of claims 1 or 2, characterized in that three of R ¹ , R ² , R ³ and R ⁴ are H.

6. A compound for use according to any one of claims 1 or 2, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is not H.

7. A compound for use according to any one of claims 1 to 6, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is C _1-3 alkyl, optionally substituted with:

hydroxy,

C _1-2 alkoxy, optionally substituted with one or more fluoro groups,

_NH2 ,

phenyl,

C _5-6 heteroaryl,

C _1-4 alkylcarbamoyl,

acylamido or

one or more fluoro groups.

8. A compound for use according to claim 7, characterized in that the C _1-3 alkyl group is unsubstituted.

9. A compound for use according to claim 7, characterized in that the C _1-3 alkyl group is perfluorinated.

10. A compound for use according to claim 7, characterized in that the C _1-3 alkyl group is substituted by a group selected from:

(i) hydroxy;

(ii) C _1-2 alkoxy;

(iii) _NH2 ;

(iv) phenyl;

(v) C _5-6 heteroaryl;

(vi) C _1-4 alkylcarbamoyl; and

(vii) acylamido.

11. A compound for use according to any one of claims 1 to 10, wherein at least one of R ¹ , R ² , R ³ and R ⁴ is C _1-3 alkoxy optionally substituted with C _3-6 cycloalkyl or one or more fluoro groups.

12. A compound for use according to claim 11, characterized in that the C _1-3 alkoxy group is unsubstituted.

13. A compound for use according to claim 11, characterized in that the C _1-3 alkoxy group is perfluorinated.

14. A compound for use according to claim 11, characterized in that the C _1-3 alkoxy group is substituted by C _3-6 cycloalkyl.

15. A compound for use according to any one of claims 1 to 14, wherein at least one of R ¹ , R ² , R ³ and R ⁴ is C _3-6 cycloalkyl.

16. A compound for use according to claim 15, characterized in that the C _3-6 cycloalkyl group is cyclopropyl.

17. A compound for use according to any one of claims 1 to 16, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is a halogen.

18. A compound for use according to any one of claims 1 to 17, wherein at least one of R ¹ , R ² , R ³ and R ⁴ is COR ^C , wherein R ^C is selected from NR ^N1 R ^N2 , wherein R ^N1 and R ^N2 are independently selected from H and methyl.

19. A compound for use according to any one of claims 1 to 18, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is selected from cyano, NH ₂ and NO ₂ .

20. A compound for use according to claim 19, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is cyano.

21. A compound for use according to claim 19, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is NH ₂ .

22. A compound for use according to claim 19, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is NO ₂ .

23. A compound for use according to any one of claims 1 to 22, wherein at least one of R ¹ , R ² , R ³ and R ⁴ is phenyl or C _5-6 heteroaryl optionally substituted with methyl, cyano, hydroxy or methoxy.

24. A compound for use according to claim 23, wherein at least one of R ¹ , R ² , R ³ and R ⁴ is phenyl, optionally substituted with methyl or methoxy.

25. A compound for use according to claim 23, wherein at least one of R ¹ , R ² , R ³ and R ⁴ is C _5-6 heteroaryl optionally substituted with one or more methyl groups.

26. A compound for use according to claim 1, wherein R ⁴ is methoxy, R ² is CH ₂ OCH ₃ or CH ₂ OCH ₂ CH ₃ , and R ¹ and R ³ are H.

27. A compound for use according to claim 1, wherein R ⁴ is methoxy, R ² is phenyl optionally substituted with methyl or methoxy, and R ¹ and R ³ are H.

28. A compound for use according to claim 1, wherein R ⁴ is methoxy, R ² is C _5-6 heteroaryl optionally substituted with methyl.

29. A compound for use according to claim 1, wherein R ⁴ is methoxy, R ² is C ₆ heteroaryl optionally substituted with methyl.

30. A compound for use according to claim 1, wherein R ⁴ is methoxy and R ¹ , R ² and R ³ are H.

31. A compound for use according to claim 1, wherein R ⁴ is chlorine, R ² is C _1-3 alkyl or bromine, and R ¹ and R ³ are H.

32. A compound for use according to claim 1, characterized in that R ⁴ is chlorine, and R ¹ , R ² and R ⁴ are H.

33. A compound for use according to claim 1, wherein R ³ is C _1-3 alkyl, and R ¹ , R ² and R ⁴ are H.

34. A compound for use according to any one of claims 1 to 33, wherein Ar is phenyl, which may be unsubstituted or substituted.

35. A compound for use according to any one of claims 1 to 33, wherein Ar is naphthyl, which may be unsubstituted or substituted.

36. A compound for use according to any one of claims 1 to 33, wherein Ar is substituted phenyl.

37. A compound for use according to any one of claims 1 to 33, wherein Ar is a C _5-10 heteroaryl group which may be unsubstituted or substituted.

38. A compound for use according to claim 37, wherein the C _5-10 heteroaryl group is selected from: quinolinyl, benzothiazolyl, quinoxalinyl, benzooxadiazolyl, benzothiadiazolyl, benzofuran and benzotriazolyl.

39. A compound for use according to claim 37, wherein the C _5-10 heteroaryl group is quinolinyl or benzothiazolyl.

40. A compound for use according to any one of paragraphs 1-33, characterized in that Ar is a group:

.

41. A compound for use according to any one of claims 33 to 39, wherein Ar is substituted by C _1-4 alkyl optionally substituted by hydroxide, C _1-2 alkoxy, NH ₂ , C _1-4 alkylcarbamoyl or one or more fluoro groups.

42. A compound for use according to claim 41, characterized in that the C _1-4 alkyl group is unsubstituted.

43. A compound for use according to claim 41, characterized in that the C _1-4 alkyl group is perfluorinated.

44. A compound for use according to claim 41, characterized in that the C _1-4 alkyl group is substituted by a group selected from:

(i) hydroxy;

(ii) C _1-2 alkoxy;

(iii) NH ₂ ; and

(iv) C _1-4 alkylcarbamoyl.

45. A compound for use according to any one of paragraphs 33-39, characterized in that Ar is substituted by C _3-6 cycloalkyl.

46. A compound for use according to claim 45, characterized in that the C _3-6 cycloalkyl group is cyclohexyl.

47. A compound for use according to any one of claims 33 to 39, wherein Ar is substituted by hydroxy; cyano; NR ^N3 R ^N4 , where R ^N3 and R ^N4 are independently selected from H and methyl; or acylamido.

48. A compound for use according to claim 47, wherein the substituent is hydroxy.

49. A compound for use according to claim 47, wherein the substituent is cyano.

50. A compound for use according to claim 47, wherein the substituent is NR ^N3 R ^N4 , wherein R ^N3 and R ^N4 are independently selected from H and methyl.

51. A compound for use according to claim 47, characterized in that the substituent is acylamido.

52. A compound for use according to any one of paragraphs 33-39, characterized in that Ar is substituted by a halogen.

53. A compound for use according to any one of claims 33 to 39, wherein Ar is substituted by C _1-3 alkoxy optionally substituted by hydroxy, C(O)NH ₂ , C _3-6 cycloalkyl, phenyl, C _5-6 heteroaryl or one or more fluoro groups.

54. A compound for use according to claim 53, characterized in that the C _1-3 alkoxy group is unsubstituted.

55. A compound for use according to claim 53, characterized in that the C _1-3 alkoxy group is perfluorinated.

56. A compound for use according to claim 53, characterized in that the C _1-3 alkoxy group is substituted by hydroxy.

57. A compound for use according to claim 53, characterized in that the C _1-3 alkoxy group is substituted with C(O)NH ₂ .

58. A compound for use according to claim 53, characterized in that the C _1-3 alkoxy group is substituted by C _3-6 cycloalkyl.

59. A compound for use according to claim 53, characterized in that the C _1-3 alkoxy group is substituted by phenyl.

60. A compound for use according to claim 53, characterized in that the C _1-3 alkoxy group is substituted by a C _5-6 heteroaryl.

61. A compound for use according to any one of claims 33 to 39, wherein Ar is substituted by phenoxy, optionally substituted by fluorine.

62. A compound for use according to claim 61, characterized in that Ar is substituted by phenoxy.

63. A compound for use according to claim 61, characterized in that Ar is substituted by OC ₆ H ₄ F.

64. A compound for use according to any one of claims 33-39, characterized in that Ar is substituted by phenyl or C _5-6 heteroaryl.

65. A compound for use according to claim 64, characterized in that Ar is substituted by phenyl.

66. A compound for use according to claim 64, characterized in that Ar is substituted with C _5-6 heteroaryl.

67. A compound for use according to any one of paragraphs 33-39, characterized in that Ar is replaced by SF ₅ or SO ₂ CH ₃ .

68. A compound for use according to claim 67, characterized in that Ar is replaced by SF ₅ .

69. A compound for use according to claim 67, characterized in that Ar is replaced by SO ₂ CH ₃ .

70. A compound for use according to any one of claims 33 to 39, wherein Ar is substituted with -(CH ₂ ) _n -Y-, where Y is O or CH ₂ , and n is 2 or 3.

71. A compound for use according to claim 70, wherein Ar is phenyl.

72. A compound for use according to any one of paragraphs 33-39, characterized in that Ar is substituted by a C _1-4 alkyl ester.

73. A compound for use according to claim 72, wherein Ar is replaced by C(O)OCH ₃ .

74. A compound for use according to claim 72, wherein Ar is replaced by C(O)OC(CH ₃ ) ₃ .

75. A compound for use according to any one of paragraphs 1-33 and 41-74, characterized in that Ar is represented by the formula (Ar-1):

(Ar-1)

where Y is N or CR ^A4 , and Z is N or CR ^A5 ; and

R ^A1 , R ^A2 , R ^A3 , R ^A4 (if present) and R ^A5 (if present) are independently selected from H and optional substituents Ar.

76. A compound for use according to claim 75, wherein R ^A2 is ethyl.

77. A compound for use according to claim 75, wherein R ^A3 is selected from cycloalkyl; phenoxy; phenyl; C _5-6 heteroaryl; SF ₅ ; and SO ₂ CH ₃ .

78. A compound for use according to any one of claims 1 to 33, wherein Ar is 5-ethyl-2-methoxyphenyl.

79. A compound for use according to any one of claims 1 to 33, wherein Ar is 5-CF ₃ -2-methoxyphenyl.

80. A compound for use according to any one of claims 1 to 33, wherein Ar is 2,6-dimethoxyphenyl.

81. A compound for use according to claim 1, provided that when:

R ¹ , R ² , R ³ and R ⁴ represent H,

Ar is not 4-aminophenyl.

82. A pharmaceutical composition comprising a compound according to the definition provided in any of paragraphs 1-81 and a pharmaceutically acceptable excipient.

83. A method for treating an oncological disease, comprising administering to a patient in need of treatment a compound according to the definition presented in any of paragraphs 1-81, or a pharmaceutical composition according to paragraph 82.

84. The method according to claim 83, characterized in that the compound is administered simultaneously or sequentially with radiation therapy and/or chemotherapy.

85. Use of a compound according to the definition provided in any of paragraphs 1-81 in the manufacture of a medicinal product for the treatment of an oncological disease.

86. A compound according to the definition provided in any of claims 1-81, or a pharmaceutical composition according to claim 82 for use in the treatment of an oncological disease.

87. A compound or pharmaceutical composition according to claim 86, characterized in that the treatment is intended for simultaneous or sequential use with radiation therapy and/or chemotherapy.

88. A compound according to the definition provided in any of paragraphs 1-81, or a pharmaceutical salt thereof.

89. The compound according to claim 88, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is not H.

90. The compound according to claim 89, characterized in that R ³ does not represent CF ₃ .

91. The compound according to claim 89, characterized in that R ³ is not substituted C _1-3 alkyl.

92. The compound according to claim 89, characterized in that R ³ is ethyl or propyl.

93. A compound according to claim 89, wherein R ³ is not C _1-3 alkyl optionally substituted by hydroxy, C _1-2 alkoxy, NH ₂ , phenyl, C _5-6 heteroaryl, C _1-4 alkylcarbamoyl, acylamido or one or more fluoro groups.

94. The compound according to claim 88, characterized in that R ⁴ is methoxy.

95. The compound according to claim 88, characterized in that R ⁴ is Cl, and R ¹ , R ² and R ³ are H.

96. The compound according to claim 88, characterized in that R ⁴ is Cl, and R ² is C _1-3 alkyl or bromine, and R ¹ and R ³ are H.

97. The compound according to claim 88, characterized in that R ³ is C _1-3 alkyl, and R ¹ , R ² and R ⁴ are H.

98. The connection according to item 88, provided that when:

R ¹ , R ² and R ³ are H and R ⁴ is methoxy,

Ar is not unsubstituted naphthyl.

99. The connection according to item 88, provided that when:

R ¹ , R ² , R ³ and R ⁴ represent H,

Ar is not 2,4,6-trimethylphenyl.

100. The connection according to paragraph 88, provided that when:

R ¹ , R ² and R ⁴ are H and R ³ is CF ₃ ,

Ar is not 2-(difluoromethoxy)phenyl.

101. The connection according to paragraph 88, provided that when:

R ¹ , R ² , R ³ and R ⁴ represent H,

Ar is not 4-fluoro-3-methylphenyl.

102. A compound for use according to paragraph 88, provided that when:

R ¹ , R ² , R ³ and R ⁴ represent H,

Ar is not 4-aminophenyl.

103. A method for synthesizing a compound according to the definition provided in any of paragraphs 88-102.

104. A compound for use according to claim 1, characterized in that:

R ¹ , R ² , R ³ and R ⁴ are independently selected from:

(i) H;

(ii) C _1-3 alkyl optionally substituted with: hydroxy, C _1-2 alkoxy, NH ₂ , phenyl, C _5-6 heteroaryl, C _1-4 alkylcarbamoyl, acylamido or one or more fluoro groups;

(iii) C _1-3 alkoxy optionally substituted with C _3-6 cycloalkyl or one or more fluoro groups;

(iv) C _3-6 cycloalkyl;

(v) halogen;

(vi) COR ^C , where R ^C is selected from NR ^N1 R ^N2 , where R ^N1 and R ^N2 are independently selected from H and methyl;

(vii) cyano, NH ₂ , NO ₂ ;

(viii) phenyl or C _5-6 heteroaryl optionally substituted with methyl, hydroxy or methoxy;

Ar is a phenyl, naphthyl or C _5-10 heteroaryl group, which groups are optionally substituted with one or more groups selected from:

(i) C _1-4 alkyl optionally substituted with hydroxy, C _1-2 alkoxy, NH ₂ , C _1-4 alkylcarbamoyl or one or more fluoro groups;

(ii) C _3-6 cycloalkyl;

(iii) hydroxy; cyano; NR ^N3 R ^N4 , where R ^N3 and R ^N4 are independently selected from H and methyl; acylamido;

(iv) halogen;

(v) C _1-3 alkoxy optionally substituted with hydroxy, C(O)NH ₂ , C _3-6 cycloalkyl, phenyl, C _5-6 heteroaryl or one or more fluoro groups;

(vi) phenoxy optionally substituted with fluorine;

(vii) phenyl, C _5-6 heteroaryl;

(viii) SF ₅ , SO ₂ CH ₃ ;

(ix) -(CH ₂ ) _n -Y-, where Y is O or CH ₂ and n is 2 or 3.

105. A compound for use according to claim 104, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is not H.

106. A compound for use according to any one of claims 104 or 105, wherein at least one of R ¹ , R ² , R ³ and R ⁴ is C _1-3 alkyl optionally substituted with: hydroxy, C _1-2 alkoxy, NH ₂ , phenyl, C _5-6 heteroaryl, C _1-4 alkylcarbamoyl, acylamido or one or more fluoro groups.

107. A compound for use according to any one of paragraphs 104-106, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is C _1-3 alkoxy optionally substituted with C _3-6 cycloalkyl or one or more fluoro groups.

108. A compound for use according to any one of paragraphs 104-107, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is C _3-6 cycloalkyl.

109. A compound for use according to any one of paragraphs 104-108, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is a halogen.

110. A compound for use according to any one of claims 104 to 109, wherein at least one of R ¹ , R ² , R ³ and R ⁴ is COR ^C , wherein R ^C is selected from NR ^N1 R ^N2 , wherein R ^N1 and R ^N2 are independently selected from H and methyl.

111. A compound for use according to any one of paragraphs 104-110, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is selected from cyano, NH ₂ and NO ₂ .

112. A compound for use according to any one of paragraphs 104-111, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is phenyl or C _5-6 heteroaryl optionally substituted with methyl, hydroxy or methoxy.

113. A connection for use according to paragraph 104, characterized in that:

(a) R ⁴ is methoxy, R ² is CH ₂ OCH ₃ or CH2OCH ₂ CH ₃ and R ¹ and R ³ are H;

(b) R ⁴ is methoxy, R ² is phenyl optionally substituted with methyl or methoxy, and R ¹ and R ³ are H;

(c) R ⁴ is methoxy, R ² is C _5-6 heteroaryl optionally substituted with methyl;

(d) R ⁴ is methoxy, and R ¹ , R ² and R ³ are H;

(e) R ⁴ is chlorine, R ² is C _1-3 alkyl or bromo, and R ¹ and R ³ are H;

(f) R ⁴ is chlorine and R ¹ , R ² and R ⁴ are H; or

(g) R ³ is C _1-3 alkyl, and R ¹ , R ² and R ⁴ are H.

114. A compound for use according to any one of paragraphs 104-113, characterized in that Ar is:

(a) phenyl, which may be substituted or unsubstituted;

(b) naphthyl, which may be substituted or unsubstituted; or

(c) a C _5-10 heteroaryl group which may be substituted or unsubstituted.

115. A compound for use according to claim 114, wherein Ar is substituted by C _1-4 alkyl optionally substituted by hydroxy, C _1-2 alkoxy, NH ₂ , C _1-4 alkylcarbamoyl or one or more fluoro groups.

116. A compound for use according to claim 114, characterized in that Ar is substituted with C _3-6 cycloalkyl.

117. A compound for use according to claim 114, wherein Ar is substituted by hydroxy; cyano; NR ^N3 R ^N4 , where R ^N3 and R ^N4 are independently selected from H and methyl; or acylamido.

118. A compound for use according to claim 114, characterized in that Ar is substituted by a halogen.

119. A compound for use according to claim 114, wherein Ar is substituted by C _1-3 alkoxy optionally substituted by hydroxy, C(O)NH ₂ , C _3-6 cycloalkyl, phenyl, C _5-6 heteroaryl or one or more fluoro groups.

120. A compound for use according to claim 114, wherein Ar is substituted by phenoxy, optionally substituted by fluorine.

121. A compound for use according to claim 114, characterized in that Ar is substituted by phenyl or C _5-6 heteroaryl.

122. A compound for use according to claim 114, characterized in that Ar is replaced by SF ₅ or SO ₂ CH ₃ .

123. A compound for use according to claim 114, wherein Ar is substituted with -(CH ₂ ) _n -Y-, where Y is O or CH ₂ , and n is 2 or 3.

124. A compound for use according to any one of paragraphs 104-113, characterized in that Ar is represented by the formula (Ar-1):

(Ar-1)

where Y is N or CR ^A4 , and Z is N or CR ^A5 ; and

R ^A1 , R ^A2 , R ^A3 , R ^A4 (if present) and R ^A5 (if present) are independently selected from H and optional substituents Ar.

125. A connection for use according to paragraph 124, characterized in that:

(a) R ^A2 is ethyl; or

(b) R ^A3 is selected from cycloalkyl; phenoxy; phenyl; C _5-6 heteroaryl; SF ₅ and SO ₂ CH ₃ .

126. A compound for use according to any one of paragraphs 104-113, characterized in that Ar is:

(a) 5-ethyl-2-methoxyphenyl;

(b) 5-CF ₃ -2-methoxyphenyl; or

(c) 2,6-dimethoxyphenyl.

127. A compound according to the definition provided in any of paragraphs 104-126, or a pharmaceutical salt thereof.

128. The connection according to item 127, characterized in that:

(a) at least one of R ¹ , R ² , R ³ and R ⁴ is not H;

(b) R ³ is not CF ₃ ;

(c) R ³ is not substituted C _1-3 alkyl;

(d) R ³ is ethyl or propyl;

(e) R ³ is not C _1-3 alkyl optionally substituted with hydroxy, C _1-2 alkoxy, NH ₂ , phenyl, C _5-6 heteroaryl, C _1-4 alkylcarbamoyl, acylamido, or one or more fluoro groups;

(f) R ⁴ is methoxy;

(g) R ⁴ is Cl, and R ¹ , R ² and R ³ are H;

(h) R ⁴ is Cl, and R ² is C _1-3 alkyl or bromo, and R ¹ and R ³ are H; or

(i) R ³ is C _1-3 alkyl, and R ¹ , R ² and R ⁴ are H.

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Claims (69)

1. Compound of formula (I) (I) or a pharmaceutically acceptable salt thereof, wherein: R ¹ , R ² , R ³ and R ⁴ are independently selected from: (i) H; (ii) C _1-3 alkyl, optionally substituted by: hydroxy, C _1-2 alkoxy, _NH2 , phenyl, C _5-6 heteroaryl, C _1-4 alkylcarbamoyl, or acylamido; (iii) C _1-3 alkoxy optionally substituted with C _3-6 cycloalkyl or one or more fluoro groups; (iv) C _3-6 cycloalkyl; (v) halogen; (vi) COR ^C , where R ^C is selected from NR ^N1 R ^N2 , where R ^N1 and R ^N2 are independently selected from H and methyl; (vii) cyano, NH ₂ or NO ₂ ; and (viii) phenyl or C _5-6 heteroaryl optionally substituted with methyl, cyano, hydroxy or methoxy; Ar is phenyl optionally substituted with one or more groups selected from: (i) C _1-4 alkyl optionally substituted with hydroxy, C _1-2 alkoxy, NH ₂ , C _1-4 alkylcarbamoyl or one or more fluoro groups; (ii) C _3-6 cycloalkyl; (iii) hydroxy; cyano; NR ^N3 R ^N4 , where R ^N3 and R ^N4 are independently selected from H and methyl; or acylamido; (iv) halogen; (v) C _1-3 alkoxy optionally substituted with hydroxy, C(O)NH ₂ , C _3-6 cycloalkyl, phenyl, C _5-6 heteroaryl or one or more fluoro groups; (vi) phenoxy optionally substituted with fluorine; (vii) phenyl or C _5-6 heteroaryl; or (viii) SF ₅ or SO ₂ CH ₃ ; and , where at least one of R ¹ , R ² , R ³ and R ⁴ is not H and wherein the C _5-6 heteroaryl has from 5 to 6 ring atoms, of which from 1 to 3 ring atoms are heteroatoms selected from nitrogen, oxygen or sulfur. 2. A compound or salt for use according to claim 1, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is C _1-3 alkyl, optionally substituted with: hydroxy, C _1-2 alkoxy, _NH2 , phenyl, C _5-6 heteroaryl, C _1-4 alkylcarbamoyl, or acylamido. 3. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 and 2, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is C _1-3 alkoxy optionally substituted with C _3-6 cycloalkyl or one or more fluoro groups. 4. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that at least one of R ¹ , R ² , R ³ and R ⁴ is a) C _3-6 cycloalkyl; b) COR ^C , wherein R ^C is selected from NR ^N1 R ^N2 , wherein R ^N1 and R ^N2 are independently selected from H and methyl; c) cyano, NH ₂ or NO ₂ ; d) phenyl or C _5-6 heteroaryl optionally substituted with methyl, hydroxy or methoxy. 5. A compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: (a) R ⁴ is methoxy, R ² is CH ₂ OCH ₃ or CH ₂ OCH ₂ CH ₃ , and R ¹ and R ³ are H; (b) R ⁴ is methoxy, R ² is phenyl optionally substituted with methyl or methoxy, and R ¹ and R ³ are H; (c) R ⁴ is methoxy, R ² is C _5-6 heteroaryl optionally substituted with methyl; (d) R ⁴ is methoxy, and R ¹ , R ² and R ³ are H; (e) R ⁴ is chlorine, R ² is C _1-3 alkyl or bromine, and R ¹ and R ³ are H; or (f) R ³ is C _1-3 alkyl, and R ¹ , R ² and R ⁴ are H. 6. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein Ar is substituted by C _1-4 alkyl, optionally substituted by hydroxy, C _1-2 alkoxy, NH ₂ , C _1-4 alkylcarbamoyl or one or more fluoro groups. 7. A compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, characterized in that Ar is substituted by C _1-3 alkoxy, optionally substituted by hydroxy, C(O)NH ₂ , C _3-6 cycloalkyl, phenyl, C _5-6 heteroaryl or one or more fluoro groups. 8. A compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs 1-5, characterized in that Ar is substituted a) C _3-6 cycloalkyl; b) hydroxy; c) cyano; d) NR ^N3 R ^N4 , where R ^N3 and R ^N4 are independently selected from H and methyl; or acylamido; e) phenoxy, optionally substituted with fluorine; f) phenyl or C _5-6 heteroaryl; g) SF ₅ or SO ₂ CH ₃ . 9. A compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs 1-5, characterized in that Ar is: (a) 5-ethyl-2-methoxyphenyl; (b) 5-CF ₃ -2-methoxyphenyl; or (c) 2,6-dimethoxyphenyl. 10. A pharmaceutical composition inhibiting lysine acetyltransferase (KAT), comprising an effective amount of a compound according to any one of claims 1-9 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 11. A compound or salt according to any one of claims 1 to 9 for use in a method of therapy for inhibiting lysine acetyltransferase (KAT). 12. A compound or salt according to any one of claims 1 to 9 for use in the treatment of cancer associated with inhibition of lysine acetyltransferase (KAT). 13. The compound or salt for use according to claim 12, wherein the cancer is selected from leukemia, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, Hodgkin's disease, prostate cancer, lung cancer, melanoma, breast cancer, colorectal cancer, colon cancer, squamous cell carcinoma, and gastric cancer. 14. A compound or salt for use according to claim 12, wherein the compound is administered simultaneously or sequentially with radiation therapy and/or chemotherapy.