HK1240585B - Substituted dihydroisoquinolinone compounds - Google Patents

Description

Substituted dihydroisoquinolinone compounds

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority to U.S. Provisional Application No. 62/013,410, filed on June 17, 2014, and U.S. Provisional Application No. 62/156,533, filed on May 4, 2015, the entire contents of each of which are incorporated herein by reference.

Field of the Invention

The present invention relates to compounds of Formulas I, I', II, II', and III, and pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising the compounds and salts, and uses thereof. The compounds, salts, and compositions of the present invention can be used to treat or ameliorate abnormal cell proliferative disorders (e.g., cancer).

background

Epigenetic alterations play a crucial role in cellular processes, including cell proliferation, differentiation, and survival. Epigenetic silencing of tumor suppressor genes and activation of oncogenes can occur through altered CpG island methylation patterns, histone modifications, and dysregulation of DNA-binding proteins. The Drosophila Pc gene represents one of a group of epigenetic effectors. EZH2 (enhancer of zeste homolog 2) is the catalytic component of the Drosophila Pc gene repressor complex 2 (PRC2), a conserved multi-subunit complex that represses gene transcription by methylating lysine 27 (H3K27) on histone H3. EZH2 plays a key role in regulating gene expression patterns, which regulate cell fate decisions such as differentiation and self-renewal. EZH2 is overexpressed in certain cancer cells and has been linked to cell proliferation, invasion, chemoresistance, and metastasis.

High EZH2 expression is associated with poor prognosis, high cancer grade, and high cancer stage in several cancer types, including breast, colorectal, endometrial, gastric, liver, kidney, lung, melanoma, ovarian, pancreatic, prostate, and bladder cancers. See Crea et al., Crit. Rev. Oncol. Hematol. 2012, 83:184-193, and references cited therein; see also Kleer et al., Proc. Natl. Acad. Sci. USA 2003, 100:11606-11; Mimori et al., Eur. J. Surg. Oncol. 2005, 31:376-80; Bachmann et al., J. Clin. Oncol. 2006, 24:268-273; Matsukawa et al., Cancer Sci. 2006, 97:484-491; Sasaki et al., Lab. Invest. 2008, 88:873-882; Sudo et al., Br. J. Cancer 2005, 92(9):1754-1758; Breuer et al., Neoplasia 2004, 6:736-43; Lu et al., Cancer Res. 2007, 67:1757-1768; Ougolkov et al., Clin. Cancer Res. 2008, 14:6790-6796; Varambally et al., Nature 2002, 419:624-629; Wagener et al., Int. J. Cancer 2008, 123:1545-1550; and Weikert et al., Int. J. Mol. Med. 2005, 16:349-353.

Recurrent somatic mutations of EZH2 have been identified in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL). Mutations altering EZH2 tyrosine 641 (e.g., Y641C, Y641F, Y641N, Y641S, and Y641H) have been reported in up to 22% of germinal center B-cell DLBCL and 7% of FL. Morin et al. Nat. Genetics 2010 Feb;42(2):181-185. Mutations at alanine 677 (A677) and alanine 687 (A687) have also been reported. McCabe et al. Proc. Natl. Acad. Sci. USA 2012, 109:2989-2994; Majer et al. FEBS Letters 2012, 586:3448-3451. It has been proposed that EZH2 activating mutations alter substrate specificity, which results in high levels of trimethylated H3K27 (H3K27me3).

Therefore, compounds that inhibit the activity of wild-type and/or mutant forms of EZH2 may be of interest for the treatment of cancer.

Overview

The present invention provides, in part, novel compounds and pharmaceutically acceptable salts. The compounds can modulate the activity of EZH2, thereby achieving biological functions, such as by inhibiting cell proliferation and invasion, inhibiting metastasis, inducing apoptosis, or inhibiting angiogenesis. The present invention also provides pharmaceutical compositions and medicaments comprising the compounds or salts of the present invention, alone or in combination with other therapeutic agents or palliatives. The present invention also provides, in part, methods for preparing the novel compounds, salts, and compositions thereof, and methods for using the novel compounds, salts, and compositions thereof.

In one aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof,

in:

R ¹ is selected from the group consisting of H, F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C(O)R ⁵ , C ₃ -C ₈ cycloalkyl, 3 to 12-membered heterocyclyl, and 5 to 12-membered heteroaryl, wherein each of the C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy is optionally substituted with one or more R ⁶ , and each of the C ₃ -C ₈ cycloalkyl, 3 to 12-membered heterocyclyl, or 5 to 12-membered heteroaryl is optionally substituted with one or more R ⁷ ;

R ² is H, F or C ₁ -C ₄ alkyl;

L is a bond or C ₁ -C ₄ alkylene;

R ³ is selected from the group consisting of C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, OH, CN, C(O)R ⁸ , COOR ⁹ , NR ¹⁰ R ¹¹ , OR ¹² , C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, and 5 to 12 membered heteroaryl, wherein each of the C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy is optionally substituted with one or more R ⁶ , and each of the C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl is optionally substituted with one or more R ⁷ ;

R ⁴ is H, halogen or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl is optionally substituted with one or more R ⁶ ;

R ⁵ is C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

each R ⁶ is independently OH, F, CN or C ₁ -C ₄ alkoxy;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or a 3- to 6-membered heterocyclyl;

R ⁸ is C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

R ⁹ is H or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

R ¹⁰ and R ¹¹ are independently H or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl is optionally substituted with one or more R ¹⁴ ;

R ¹² is selected from the group consisting of C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, and 5 to 12 membered heteroaryl, wherein said C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl is each optionally substituted with one or more R ⁷ ;

each R ¹³ is independently C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

R ¹⁴ and R ¹⁵ are each independently OH, F, CN or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In some embodiments, the compound of Formula (I) has the absolute stereochemistry at the carbon atom bearing the R ¹ and R ² substituents, as shown in Formula (IA) or (IB):

or a pharmaceutically acceptable salt thereof,

in:

R ¹ , R ² , L, R ³ , R ⁴ , X and Z have the same meanings as defined for formula (I).

In another aspect, the present invention provides a compound of formula (II), (II-A) or (II-B), or a pharmaceutically acceptable salt thereof,

in:

R ¹ , L, R ³ and X have the same meanings as defined for formula (I); and

R ⁴ is H, Cl, Br, F or CH ₃ .

In another aspect, the present invention provides a compound of formula (III), or a pharmaceutically acceptable salt thereof,

in:

R ¹ and R ³ are taken together to form a 3- to 12-membered heterocyclyl group optionally substituted with one or more R ⁷ ;

R ² is H, F or C ₁ -C ₄ alkyl;

R ⁴ is H, halogen or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl is optionally substituted with one or more R ⁶ ;

R ⁶ are each independently OH, F, CN or C ₁ -C ₄ alkoxy;

R ⁷ is each independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ or a 3- to 6-membered heterocyclyl;

R ¹³ is each independently C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

R ¹⁵ is each independently OH, F, CN or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of one of the formulae described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. In some embodiments, the pharmaceutical composition comprises two or more pharmaceutically acceptable carriers and/or excipients.

The present invention also provides methods of treatment and uses, which comprise administering the compound of the present invention or a pharmaceutically acceptable salt thereof.

In one aspect, the present invention provides a method for treating abnormal cell growth in a patient, comprising administering to the patient a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof.

In common embodiments of the methods provided herein, the abnormal cell growth is cancer. In some embodiments, the methods provided herein result in one or more of the following effects: (1) inhibiting cancer cell proliferation; (2) inhibiting cancer cell invasion; (3) inducing cancer cell apoptosis; (4) inhibiting cancer cell metastasis; or (5) inhibiting angiogenesis.

In another aspect, the present invention provides a method for treating an EZH2-mediated disorder in a patient, comprising administering to the patient a compound of the present invention or a pharmaceutically acceptable salt thereof in an amount effective to treat the disorder. The compounds and salts of the present invention can inhibit wild-type and certain mutant human histone methyltransferase EZH2.

In another aspect, the present invention provides a compound of one of the formulae described herein, or a pharmaceutically acceptable salt thereof, for use in treating abnormal cell growth in a patient.

In another aspect, the present invention provides the use of a compound of one of the formulae described herein, or a pharmaceutically acceptable salt thereof, for treating abnormal cell growth in a patient.

In another aspect, the present invention provides the use of a compound of one of the formulae described herein or a pharmaceutically acceptable salt thereof for preparing a medicament for treating abnormal cell growth.

In common embodiments, the abnormal cell growth is cancer and the patient is a human.

In some embodiments, the methods described herein further comprise administering to the patient an amount of an anticancer therapeutic or mitigating agent that is effective together to treat the abnormal cell growth. In some embodiments, the one or more anticancer therapeutic agents are selected from anti-tumor agents, anti-angiogenic agents, signal transduction inhibitors, and anti-proliferative agents, and the amounts are effective together to treat the abnormal cell growth. In some such embodiments, the anti-tumor agent is selected from the group consisting of: mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, radiation, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxic agents, anti-hormones, and anti-androgens.

In other embodiments, the uses described herein include the use of a compound of one of the formulae described herein or a pharmaceutically acceptable salt thereof in combination with one or more substances selected from anti-tumor agents, anti-angiogenic agents, signal transduction inhibitors and anti-proliferative agents.

In some embodiments, the drugs described herein may be suitable for use in combination with one or more agents selected from anti-tumor agents, anti-angiogenic agents, signal transduction inhibitors, and anti-proliferative agents.

Each embodiment of the compounds of the present invention described below can be combined with one or more other embodiments of the compounds of the present invention described herein (where such other embodiments are not inconsistent with the embodiments with which they are combined). In addition, the following descriptions of the various embodiments of the present invention contemplate the inclusion within their scope of pharmaceutically acceptable salts of the compounds of the present invention. Therefore, the phrase "or a pharmaceutically acceptable salt thereof" is implicit in the descriptions of all compounds described in this application.

Detailed description

The present invention can be more readily understood with reference to the following detailed description of preferred embodiments of the present invention and the included Examples. It should be understood that the terms used in this application are intended only to describe the purpose of specific embodiments and are not intended to limit them. It should be further understood that, unless expressly defined herein, the meaning of the terms used in this application is the customary meaning known in the relevant art.

As used herein, the singular forms "a," "an," and "the" include plural references unless otherwise indicated. For example, "a" substituent includes one or more substituents.

"Alkyl" refers to a saturated monovalent aliphatic hydrocarbon radical, including straight-chain and branched groups, having the specified number of carbon atoms. Alkyl substituents typically contain 1 to 20 carbon atoms (" _Ci - _C20 alkyl"), preferably 1 to 12 carbon atoms (" _Ci - _Ci2 alkyl"), more preferably 1 to 8 carbon atoms (" _Ci - _C8 alkyl"), or 1 to 6 carbon atoms (" _Ci - _C6 alkyl"), or 1 to 4 carbon atoms (" _Ci - _C4 alkyl"). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, n-heptyl, n-octyl, and the like. Alkyl groups may be substituted or unsubstituted. In particular, unless otherwise specified, an alkyl group may be substituted with one or more (up to the total number of hydrogen atoms present on the alkyl group) halo groups. Thus, C ₁ -C ₄ alkyl includes haloalkyl, particularly fluoroalkyl of 1 to 4 carbon atoms, for example, trifluoromethyl or difluoroethyl (ie, CF ₃ and -CH ₂ CHF ₂ ).

Alkyl groups described herein as optionally substituted may be substituted with one or more substituents, which are independently selected unless otherwise indicated. The total number of substituents may be equal to the total number of hydrogen atoms on the alkyl moiety, as long as the substitution is chemically reasonable. Optionally substituted alkyl groups typically contain 1 to 6 optional substituents, sometimes 1 to 5 optional substituents, preferably 1 to 4 optional substituents, or more preferably 1 to 3 optional substituents.

Suitable optional substituents for the alkyl group include, but are not limited to, C3- _{C8 cycloalkyl} , 3- to 12-membered heterocyclyl, _C6 - _C12 aryl, and 5- to 12-membered heteroaryl, halogen, ═O(oxo), ═S(thiocarbonyl) ^, ═N- ^CN , ═N- ^ORx , _═NRx , ^-CN , -C(O) ^Rx , _-CO2Rx , -C(O) ^NRxRy , ^-SRx , ^-SORx , -SO2Rx, ^{-SO2NRxRy ,} _{-NO2 ,} ^-NRxRy , ^-NRxC (O) ^Ry , ^-NRxC (O ^{) NRxRy} _, ^{-NRxC (} _O ^{) ORx} , ^-NRxSO2Ry , ^-NRxSO2NRxRy , ^-ORx , ^-OC (O ^{) Rx ,} and -OC(O) ^NRxR wherein ^{Rx and Ry are} each independently H, _C1 - _C8 alkyl, C1- _C8 acyl, _C2 -C8 alkenyl, _C2 - _C8 alkynyl, _C3 - _C8 cycloalkyl, 3- to ₁₂ -membered heterocyclyl, _C6 - _C12 aryl, or _5- to 12-membered heteroaryl, or ^Rx and ^Ry may be taken together with the nitrogen atom to which they are attached to form a 3- to 12-membered heterocyclyl or 5- to 12-membered heteroaryl, each optionally containing 1, 2, or 3 additional heteroatoms selected from O, N, and S; and ^Rx and ^Ry are each optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, =O, =S, =N-CN, =N-OR', =NR', -CN, -C(O)R', _-CO2R ', -C(O) _NR'2 , -SR', -SOR', _-SO2 R', -SO ₂ NR' ₂ , -NO ₂ , -NR' ₂ , -NR'C(O)R', -NR'C(O)NR' ₂ , -NR'C(O)OR', -NR'SO ₂ R', -NR'SO ₂ NR' ₂ , -OR', -OC(O)R' and -OC(O)NR'₂; wherein each R' is independently H, C ₁ -C ₈ alkyl, C ₁ -C ₈ acyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, C ₆ -C ₁₂ aryl or C ₅ -C ₁₂ heteroaryl; and wherein said C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, C ₆ -C _{The 12-} membered aryl and 5- to 12-membered heteroaryl groups are each optionally substituted as further defined herein.

Typical substituents on alkyl groups include halogen, -OH, C ₁ -C ₄ alkoxy, -OC ₆ -C ₁₂ aryl, -CN, =O, -COOR ^x , -OC(O)R ^x , -C(O)NR ^x R ^y , -NR ^x C(O)R ^y , -NR ^x R ^y , C ₃ -C ₈ cycloalkyl, C ₆ -C ₁₂ aryl, 5 to 12 membered heteroaryl and 3 to 12 membered heterocyclyl; wherein R ^x and R ^y are each independently H or C ₁ -C ₄ alkyl, or R ^x and R ^y may be taken together with the nitrogen to which they are attached to form a 3 to 12 membered heterocyclyl or 5 to 12 membered heteroaryl ring, each of which optionally contains 1, 2 or 3 additional heteroatoms selected from O, N and S; wherein the C ₃ -C ₈ cycloalkyl, C ₆ -C 12 aryl, 5 to 12 membered heteroaryl and 3 to 12 membered heterocyclyl; The _5- to 12-membered aryl, 5- to 12-membered heteroaryl, and 3- to 12-membered heterocyclyl are each optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, -OH, =O, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₄ alkoxy-C ₁ -C ₆ alkyl, -CN, -NH ₂ , -NH(C ₁ -C ₄ alkyl) and -N(C ₁ -C ₄ alkyl) ₂ .

In some embodiments, the alkyl group is optionally substituted with one or more substituents, and preferably 1 to 3 substituents, independently selected from the group consisting of halogen, -OH, C ₁ -C ₄ alkoxy, -OC ₆ -C ₁₂ aryl, -CN, ＝O, -COOR ^x , -OC(O)R x , -C(O)NR ^x R ^y , -NR x C(O)R ^y , -NR ^x R ^y , C ₃ -C ₈ cycloalkyl, C ₆ -C ₁₂ aryl, 5- ^to 12-membered heteroaryl, and 3- to 12-membered heterocyclyl; wherein R ^{x and R y are} each independently H or C ₁ -C ₄ alkyl, or R ^x and R ^y can be taken together with the N to which they are attached to form a 3- to 12-membered heterocyclyl or 5- to 12-membered heteroaryl ring, each of which optionally contains 1, 2, or 3 additional heteroatoms selected from O, N, and S; and the C ₃ -C ₈ -cycloalkyl, C ₆ -C ₁₂ aryl, 5- to 12-membered heteroaryl, and 3- to 12-membered heterocyclyl are each optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, -OH, =O, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₄ alkoxy-C ₁ -C ₆ alkyl, -CN, -NH ₂ , -NH(C ₁ -C ₄ alkyl) and -N(C ₁ -C ₄ alkyl) ₂ .

In other embodiments, the alkyl group is optionally substituted with one or more substituents, and preferably with 1 to 3 substituents, independently selected from the group consisting of halogen, -OH, C ₁ -C ₄ alkoxy, -CN, -NR ^x R ^y , C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, C ₆ -C ₁₂ aryl, and 5 to 12 membered heteroaryl; wherein R ^x and R ^y are each independently H or C ₁ -C ₄ alkyl, or R ^x and R ^y may be taken together with the N to which they are attached to form a 3 to 12 membered heterocyclyl or 5 to 12 membered heteroaryl ring, each of which optionally contains 1, 2 or 3 additional heteroatoms selected from O, N and S; and wherein the cycloalkyl, heterocyclyl, aryl or heteroaryl group is each optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, -OH, =O, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₄ alkoxy-C ₁ -C ₆ alkyl, -CN, -NH ₂ , -NH(C ₁ -C ₄ alkyl) and -N(C ₁ -C ₄ alkyl) ₂ .

In some cases, substituted alkyl groups may be specifically named based on the substituents. For example, "haloalkyl" refers to an alkyl group having the specified number of carbon atoms that is substituted with one or more halogen substituents, and typically contains 1 to 6 carbon atoms and 1, 2, or 3 halogen atoms (i.e., "C ₁ -C ₆ haloalkyl") or sometimes 1 to 4 carbon atoms and 1, 2, or 3 halogen atoms (i.e., "C ₁ -C ₄ haloalkyl"). Thus, C ₁ -C ₄ haloalkyl includes trifluoromethyl (-CF ₃ ) and difluoromethyl (-CF ₂ H). More specifically, fluorinated alkyl groups may be specifically referred to as fluoroalkyl groups, for example, C ₁ -C ₆ or C ₁ -C ₄ fluoroalkyl.

Similarly, "hydroxyalkyl" refers to an alkyl group having the specified number of carbon atoms, substituted with one or more hydroxy substituents, and typically contains 1 to 6 carbon atoms and 1, 2, or 3 hydroxy groups (i.e., " _C1 - _C6 hydroxyalkyl"). Thus, _C1 - _C6 hydroxyalkyl includes hydroxymethyl ( _-CH2OH ) and 2- _hydroxyethyl ( _-CH2CH2OH ).

"Alkoxyalkyl" refers to an alkyl group having the specified number of carbon atoms, substituted with one or more alkoxy substituents. Alkoxyalkyl groups typically contain 1 to 6 carbon atoms in the alkyl portion and are substituted with 1, 2, or 3 _C1 - _C4 alkoxy substituents. This group is sometimes referred to herein as _C1 - _C4 alkoxy- _C1 - _C6 alkyl.

"Aminoalkyl" refers to an alkyl group having the specified number of carbon atoms, substituted with one or more substituted or unsubstituted amino groups, as further defined herein. Aminoalkyl groups typically contain from 1 to 6 carbon atoms in the alkyl portion and are substituted with 1, 2, or 3 amino substituents. Thus, _C1 - _C6 aminoalkyl groups include, for example, aminomethyl ( _-CH2NH2 ), N,N- _{dimethylaminoethyl} ( _-CH2CH2N ( _CH3 ) ₂ ), 3-(N-cyclopropylamino)propyl _{( -CH2CH2CH2NH} - ^cPr ) _, and N-pyrrolidinylethyl _{( -CH2CH2} - _N -pyrrolidinyl).

"Alkenyl" refers to an alkyl group as defined herein, consisting of at least two carbon atoms and at least one carbon-carbon double bond. Typically, an alkenyl group has 2 to 20 carbon atoms (" _C2 - _C20 alkenyl"), preferably 2 to 12 carbon atoms (" _C2 - _C12 alkenyl"), more preferably 2 to 8 carbon atoms (" _C2 - _C8 alkenyl"), or 2 to 6 carbon atoms (" _C2 - _C6 alkenyl"), or 2 to 4 carbon atoms (" _C2 - _C4 alkenyl"). Representative examples include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, or 3-butenyl. An alkenyl group may be unsubstituted or substituted with groups suitable for an alkyl group as described herein.

"Alkynyl" refers to an alkyl group, as defined herein, consisting of at least two carbon atoms and at least one carbon-carbon triple bond. Alkynyl groups have 2 to 20 carbon atoms (" _C2 - _C20 alkynyl"), preferably 2 to 12 carbon atoms (" _C2 - _C12 alkynyl"), more preferably 2 to 8 carbon atoms (" _C2 - _C8 alkynyl"), or 2 to 6 carbon atoms (" _C2 - _C6 alkynyl"), or 2 to 4 carbon atoms (" _C2 - _C4 alkynyl"). Representative examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, or 3-butynyl. Alkynyl groups may be unsubstituted or substituted with the groups described herein as suitable for alkyl groups.

As used herein, "alkylene" refers to a divalent hydrocarbon radical having a specified number of carbon atoms that can link two other groups together. Sometimes, alkylene refers to a -( _CH2 ) _n- radical, where n is 1 to 8, preferably n is 1 to 4. When specified, alkylene may also be substituted with other groups and may include one or more unsaturations (i.e., alkenylene or alkynylene) or rings. The open valences of an alkylene group do not necessarily have to be at opposite ends of the chain. Thus, branched alkylene groups, such as -CH(Me)- and -C(Me) _2- , are also included within the scope of the term "alkylene," including cyclic groups such as cyclopropane-1,1-diyl and unsaturated groups such as ethylene (-CH=CH-) or propylene (-CH2 _- CH=CH-). When an alkylene group is described as optionally substituted, the substituents include those typically present on an alkyl group as described herein.

“Heteroalkylene” refers to an alkylene group as defined above, wherein one or more non-linked carbon atoms of the alkylene chain are replaced by —N(R)—, —O—, or —S(O) _q —, wherein R is H or C ₁ -C ₄ alkyl and q is 0 to 2. For example, the group —O—(CH ₂ ) _1-4 - is a “C ₂ -C ₅ ”-heteroalkylene group, wherein one of the carbon atoms of the corresponding alkylene group is replaced by O.

"Alkoxy" refers to a monovalent -O-alkyl group, wherein the alkyl portion has the specified number of carbon atoms. Alkoxy groups typically contain 1 to 8 carbon atoms (" _Ci - _Cs alkoxy"), or 1 to 6 carbon atoms (" _Ci - _C6 alkoxy"), or 1 to 4 carbon atoms ("Ci _{- C4} alkoxy"). For example, Ci- _C4 alkoxy groups include _-OCH3 , _-OCH2CH3 , _-OCH ( _CH3 ) ₂ , -OC( _CH3 ) ₃ , and the like. These groups may also be referred to herein _as methoxy, ethoxy, isopropoxy, tert-butoxy, and the like. Alkoxy groups may be unsubstituted or substituted on the alkyl portion with the groups described herein as appropriate for alkyl groups. In particular, alkoxy groups may be substituted with one or more (up to the total number of hydrogen atoms present on the alkyl portion) halo groups. Thus, C ₁ -C ₄ alkoxy includes halogenated alkoxy groups, for example, trifluoromethoxy and 2,2-difluoroethoxy (i.e., -OCF ₃ and -OCH ₂ CHF ₂ ). In some cases, a halogenated alkoxy group may be referred to as a "haloalkoxy" (or, when fluorinated, more specifically, a "fluoroalkoxy"), which has the specified number of carbon atoms and is substituted with one or more halogen substituents, and typically contains 1 to 6 carbon atoms and 1, 2, or 3 halogen atoms (i.e., "C ₁ -C ₆ haloalkoxy") or sometimes contains 1 to 4 carbon atoms and 1, 2, or 3 halogen atoms (i.e., "C ₁ -C ₄ haloalkoxy"). Thus, C ₁ -C ₄ haloalkoxy includes trifluoromethoxy (-OCF ₃ ) and difluoromethoxy (-OCF ₂ H). More specifically, a fluorinated alkoxy group may be specifically referred to as a fluoroalkoxy group, for example, a C ₁ -C ₆ or C ₁ -C ₄ fluoroalkoxy group.

Similarly, "thioalkoxy" refers to a monovalent -S-alkyl group, wherein the alkyl portion has the specified number of carbon atoms and is optionally substituted with groups described herein as suitable for alkyl groups _. For example, _C1 - _C4 thioalkoxy includes _-SCH3 and _-SCH2CH3 .

"Cycloalkyl" refers to a non-aromatic, saturated or partially unsaturated carbon ring system containing the specified number of carbon atoms, which can be a monocyclic, bridged or fused bicyclic or polycyclic ring system, which is attached to the base molecule through a carbon atom on the cycloalkyl ring. Typically, the cycloalkyl groups of the present invention contain 3 to 12 carbon atoms (" _C3 - _C12 cycloalkyl"), preferably 3 to 8 carbon atoms (" _C3 - _C8 cycloalkyl"). Representative examples include, for example, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclohexadiene, cycloheptane, cycloheptatriene, adamantane, and the like. The cycloalkyl group can be unsubstituted or substituted with the groups described herein as suitable for an alkyl group.

Examples of cycloalkyl rings include, but are not limited to

"Cycloalkylalkyl" may be used to describe a cycloalkyl ring, typically a _C3 - _C8 cycloalkyl group, which is attached to the base molecule via an alkylene linker, typically a _C1 - _C4 alkylene group. Cycloalkylalkyl groups are described by the total number of carbon atoms in the carbon ring and the linker, which typically contains 4 to 12 carbon atoms (" _C4 - _C12 cycloalkylalkyl"). Thus, cyclopropylmethyl is a _C4 -cycloalkylalkyl group, and cyclohexylethyl is a _C8 -cycloalkylalkyl group. Cycloalkylalkyl groups may be unsubstituted or substituted on the cycloalkyl and/or alkylene portions with the groups described herein as suitable for alkyl groups.

The terms "heterocyclyl,""heterocycle," or "heteroalicyclic" are used interchangeably herein to refer to a non-aromatic, saturated or partially unsaturated ring system containing the specified number of ring atoms (including at least one heteroatom selected from N, O, and S as a ring member), wherein the heterocycle is attached to the base molecule via a ring atom (which may be C or N). The heterocycle may be fused to one or more other heterocycles or carbocycles, and the fused ring may be saturated, partially unsaturated, or aromatic. Preferably, the heterocycle contains from 1 to 4 (more preferably from 1 to 2) ring heteroatoms selected from N, O, and S as ring members, provided that the heterocycle does not contain two adjacent oxygen atoms. The heterocyclyl group may be unsubstituted or substituted with the groups described herein as suitable for alkyl, aryl, or heteroaryl groups. Additionally, the ring N atoms may be optionally substituted with groups suitable for amines (e.g., alkyl, acyl, carbamoyl, sulfonyl substituents, etc.), and the ring S atoms may be optionally substituted with one or two oxo groups (i.e., S(O) _q , where q is 0, 1, or 2).

Preferred heterocycles include 3 to 12 membered heterocyclyls as defined herein.

Examples of partially unsaturated heterocyclic groups include, but are not limited to:

Examples of bridged and fused heterocyclic groups include, but are not limited to:

Examples of saturated heterocyclic groups include, but are not limited to:

In common embodiments, the heterocycle contains 3 to 12 ring members, which include carbon and non-carbon heteroatoms, preferably 4 to 6 ring members. In certain embodiments, the substituents comprising a 3 to 12-membered heterocycle are selected from the group consisting of: azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl and thiomorpholinyl, each of which may be optionally substituted as long as the substitution is chemically reasonable. In other embodiments, the substituents comprising a 3 to 12-membered heterocycle are selected from the group consisting of: oxetane, tetrahydrofuranyl and tetrahydropyranyl, each of which may be optionally substituted as long as the substitution is chemically reasonable. In specific embodiments, the 3 to 12-membered heterocycle is oxetane, which may be optionally substituted as long as the substitution is chemically reasonable.

It should be understood that generally no more than two N, O or S atoms are linked sequentially, with the exception of the case where an oxo group is linked to a N or S group to form a nitro or sulfonyl group, or certain heteroaromatic rings such as triazine, triazole, tetrazole, diazole, thiadiazole, etc.

The term "heterocycloalkyl" may be used to describe a heterocyclic group of a specified size, which is attached to the base molecule via an alkylene linker of a specified length. Typically, a heterocycloalkyl group contains an optionally substituted 3- to 12-membered heterocyclic ring attached to the base molecule via a _C1 - _C4 alkylene linker. Where indicated, the heterocycloalkyl group may be optionally substituted on the alkylene portion with groups as described herein for alkyl groups, and on the heterocyclic portion with groups as described herein for heterocycles.

"Aryl" or "aromatic group" refers to an optionally substituted monocyclic, biaryl, or fused bicyclic or polycyclic ring system having the well-known aromatic properties, wherein at least one ring contains a completely conjugated pi-electron system. Typically, an aryl group contains 6 to 20 carbon atoms (" _C6 - _C20 aryl"), preferably 6 to 14 carbon atoms (" _C6 - _C14 aryl"), or more preferably 6 to 12 carbon atoms (" _C6 - _C12 aryl") as ring members. Fused aryl groups can include an aryl ring (e.g., a benzene ring) fused to another aryl ring or to a saturated or partially unsaturated carbocyclic or heterocyclic ring. The point of attachment to the base molecule on the fused aryl ring system can be a C atom of the aromatic portion or a C or N atom of the non-aromatic portion of the ring system. Non-limiting examples of aryl groups include phenyl, biphenyl, naphthyl, anthracenyl, phenanthrenyl, indanyl, indenyl, and tetrahydronaphthyl. Aryl groups can be unsubstituted or substituted as further described herein.

Similarly, "heteroaryl" or "heteroaromatic" refers to a monocyclic, heterobiaryl, or fused bicyclic or polycyclic ring system having well-known aromaticity and containing a specified number of ring atoms and including at least one heteroatom selected from N, O, or S as a ring member in the aromatic ring. The inclusion of heteroatoms allows 5- and 6-membered rings to have aromaticity. Typically, a heteroaryl group contains 5 to 20 ring atoms ("5-20 membered heteroaryl"), preferably 5 to 14 ring atoms ("5-14 membered heteroaryl"), and more preferably 5 to 12 ring atoms ("5-12 membered heteroaryl"). The heteroaryl ring is connected to the base molecule through the ring atoms of the heteroaromatic ring to maintain aromaticity. Thus, a 6-membered heteroaryl ring can be connected to the base molecule through a C atom on the ring, while a 5-membered heteroaryl ring can be connected to the base molecule through a C or N atom on the ring. Examples of unsubstituted heteroaryl groups generally include, but are not limited to, pyrrole, furan, thiophene, pyrazole, imidazole, isoxazole, oxazole, isothiazole, thiazole, triazole, diazole, thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, pyrazine, benzofuran, benzothiophene, indole, benzimidazole, indazole, quinoline, isoquinoline, purine, triazine, naphthyridine, and carbazole. In a common preferred embodiment, the 5- or 6-membered heteroaryl group is selected from the group consisting of pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, triazolyl, pyridyl, pyrimidinyl, pyrazinyl, and pyridazinyl rings. The heteroaryl group can be unsubstituted or substituted, as further described herein.

The optionally substituted aryl, heteroaryl and heterocyclic radicals described herein may be substituted with one or more substituents, which are independently selected unless otherwise indicated. The total number of substituents may be equal to the total number of hydrogen atoms on the aryl, heteroaryl or heterocyclic radical, as long as the substitution is chemically reasonable and maintains aromaticity in the aryl and heteroaryl rings. The optionally substituted aryl, heteroaryl or heterocyclic radical typically contains 1 to 5 optional substituents, sometimes contains 1 to 4 optional substituents, preferably 1 to 3 optional substituents, or more preferably 1 to 2 optional substituents.

Suitable optional substituents for aryl, heteroaryl, and heterocyclyl rings include, but are not limited to, C1 _{- C8} alkyl, _C2 - _C8 alkenyl, _C2 - _{C8 alkynyl, C3-C8 cycloalkyl} , 3- to ₁₂ -membered heterocyclyl, _C6 - _C12 aryl _, ^and 5- to 12-membered heteroaryl; as well ^as halogen, ⁼ O, ^-CN , -C(O ₎ ^Rx , _-CO2Rx , -C ⁽ O ^{) NRxRy} , ^-SRx , ^-SORx , -SO2Rx, ^{-SO2NRxRy ,} _-NO2 , ^-NRxRy , ^-NRxC (O) ^Ry , ^-NRxC (O ₎ ^NRxRy , ^{-NRxC (} O ^{) ORx} _, ^-NRxSO2Ry , ^-NRxSO2NRxRy , ^-ORx wherein ^{Rx and Ry} are each independently H, _C1 -C8 alkyl, C1- _C8 acyl, C2-C8 alkenyl, _C2 - _C8 alkynyl, _C3 - _C8 cycloalkyl, 3- to 12 _{-membered heterocyclyl, C6-C12 aryl, or 5-} ^to _{12 -} ^membered _heteroaryl , or ^Rx and ^Ry may be taken together with the nitrogen atom to which they are attached to form a _3- to 12 _- membered heterocyclyl or 5- to 12-membered heteroaryl, each optionally containing 1, 2, or 3 additional heteroatoms selected from O, N, and S; and ^Rx and ^Ry are each optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, ＝O, ＝S, ＝N-CN, ＝N-OR′, ＝NR′, -CN, -C(O)R′, _-CO2R ′, -C(O)NR′ ₂ , -SR', -SOR', -SO ₂ R', -SO ₂ NR' ₂ , -NO ₂ , -NR' ₂ , -NR'C(O)R', -NR'C(O)NR' ₂ , -NR'C(O)OR', -NR'SO ₂ R', -NR'SO ₂ NR' ₂ , -OR', -OC(O)R' and -OC(O)NR' ₂ , wherein each R' is independently H, C ₁ -C ₈ alkyl, C ₁ -C ₈ acyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, C ₆ -C ₁₂ aryl or 5 to 12 membered heteroaryl; and the C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₃ -C 8 ₈ -cycloalkyl, 3- to 12-membered heterocyclyl, C ₆ -C ₁₂ aryl, and 5- to 12-membered heteroaryl are each optionally substituted as further defined herein.

In typical embodiments, optional substituents on the aryl, heteroaryl, and heterocyclyl rings include one or more substituents, preferably 1 to 3 substituents, independently selected from the group consisting of halogen, _C1 - _C8 alkyl, -OH, _C1 - _C8 alkoxy, CN, ＝O, -C(O) ^Rx , ^-COORx , -OC(O) ^Rx , -C(O) ^NRxRy , ^-NRxC (O)Ry, ^{-SRx , -SORx} , ^-SO2Rx , _{-SO2NRxRy ,} ^{-NO2 ,} _-NRxRy , ^-NRxC (O) _Ry ^{, -NRxC (} O) ^NRxRy , ^-NRxC (O ₎ ^ORy , ^{-NRxSO2Ry , -NRxSO2NRxRy , -OC} (O) ^{Rx ,} -OC( ^{O ) NRx} R ^y , C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, C ₆ -C ₁₂ aryl, 5 to 12 membered heteroaryl, -O-(C ₃ -C ₈ cycloalkyl), -O-(3 to 12 membered heterocyclyl), -O-(C ₆ -C ₁₂ aryl) and -O-(5 to 12 membered heteroaryl); wherein R ^x and R ^y are each independently H or C ₁ -C ₄ alkyl, or R ^x and R ^y may be taken together with the N to which they are attached to form a 3 to 12 membered heterocyclyl or 5 to 12 membered heteroaryl ring, each of which optionally contains 1, 2 or 3 additional heteroatoms selected from O, N and S; and wherein said C ₁ -C ₈ alkyl, C ₁ -C ₈ alkoxy, C 3 -C 8 cycloalkyl, 3 to 12 membered heterocyclyl, C 6 -C 12 aryl, 5 to 12 membered heteroaryl, -O-(C ₃ -C ₈ cycloalkyl), -O-(3 to 12 membered heterocyclyl), -O-(C ₆ -C ₁₂ aryl) and -O-(5 to 12 membered heteroaryl) _{-C 3} -C ₈ cycloalkyl), -O-(3- to 12-membered heterocyclyl), -O-(C ₆ -C ₁₂ aryl), and -O-(5- to 12-membered heteroaryl), which are described as optional substituents or a portion of R ^x or R ^y is optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, -OH, =O, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ hydroxyalkyl, C ₁ -C ₄ alkoxy-C ₁ -C ₆ alkyl, -CN, -NH ₂ , -NH(C ₁ -C ₄ alkyl), -N(C ₁ -C ₄ alkyl) ₂ , and N-pyrrolidinyl.

Examples of monocyclic heteroaryl groups include, but are not limited to

Examples of fused ring heteroaryl groups include, but are not limited to:

"Arylalkyl" refers to an aryl group as described herein that is attached to a base molecule via an alkylene group or a similar linker. Arylalkyl groups are described by the total number of carbon atoms in the ring and linker. Thus, benzyl is a _C7 -arylalkyl and phenethyl is a _C8 -arylalkyl. Typically, arylalkyl groups contain 7 to 16 carbon atoms (" _C7 - _C16 arylalkyl"), with the aryl portion containing 6 to 12 carbon atoms and the alkylene portion containing 1 to 4 carbon atoms. Arylalkyl groups can also be represented as _-C1 - _C4 alkylene- _C6 - _C12 aryl.

"Heteroarylalkyl" refers to a heteroaryl group as defined above, attached to the base molecule via an alkylene linker, and differs from "arylalkyl" in that at least one ring atom in the aromatic group is a heteroatom selected from N, O, and S. Heteroarylalkyl groups are sometimes described herein by the total number of non-hydrogen atoms (i.e., C, N, S, and O atoms) in the combined ring and linker (excluding substituents). Thus, for example, pyridylmethyl can be represented as " _C7 "-heteroarylalkyl. Typically, unsubstituted heteroarylalkyl groups contain 6 to 20 non-hydrogen atoms (including C, N, S, and O atoms), with the heteroaryl portion typically containing 5 to 12 atoms and the alkylene portion typically containing 1 to 4 carbon atoms. Heteroarylalkyl groups can also be represented as _-C1 - _C4alkylene -5- to 12-membered heteroaryl.

Similarly, "arylalkoxy" and "heteroarylalkoxy" refer to aryl and heteroaryl groups attached to the base molecule through a heteroalkylene linker (i.e., -O-alkylene), wherein the group is described based on the total number of non-hydrogen atoms (i.e., C, N, S, and O atoms) in the bound ring and linker. Thus, -O- _CH2 -phenyl and -O- _CH2 -pyridyl are represented as _C8 -arylalkoxy and _C8 -heteroarylalkoxy, respectively.

When an arylalkyl, arylalkoxy, heteroarylalkyl, or heteroarylalkoxy group is described as being optionally substituted, the substituents may be on the divalent linker portion of the group or on the aryl or heteroaryl portion. The substituents optionally present on the alkylene or heteroalkylene portion are generally the same as those described above for an alkyl or alkoxy group, and the substituents optionally present on the aryl or heteroaryl portion are generally the same as those described above for an aryl or heteroaryl group.

"Hydroxy" refers to an -OH group.

"Acyloxy" refers to a monovalent radical -OC(O)alkyl, wherein the alkyl portion has the specified number of carbon atoms (typically _C1 - _C8 , preferably _C1 - _C6 or _C1 - _C4 ), and is optionally substituted with groups suitable for an alkyl group. Thus, _C1 - _C4 acyloxy includes -OC(O) _C1 - _C4 alkyl substituents, for example -OC(O) _CH3 .

"Acylamino" refers to a monovalent radical, -NHC(O)alkyl or -NRC(O)alkyl, wherein the alkyl portion has the specified number of carbon atoms (typically _C1 - _C8 , preferably _C1 - _C6 or _C1 - _C4 ), and is optionally substituted with groups suitable for an alkyl group. Thus, _C1 - _C4 acylamino includes -NHC(O) _C1 - _C4 alkyl substituents, for example, -NHC(O) _CH3 .

"Aryloxy" or "heteroaryloxy" refers to an optionally substituted -O-aryl or -O-heteroaryl group, with aryl and heteroaryl being as further defined herein in each instance.

"Arylamino" or "heteroarylamino" refers to an optionally substituted -NH-aryl, -NR-aryl, -NH-heteroaryl or -NR-heteroaryl, in each case as further defined herein, and R represents a substituent suitable for an amine, such as alkyl, acyl, carbamoyl or sulfonyl.

"Cyano" refers to a -C≡N group.

"Unsubstituted amino" refers to a _-NH2 group. When an amino group is described as being substituted or optionally substituted, the term includes the form ^-NRxRy , where ^{Rx and Ry are} each independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, acyl, sulfonyl, aryl, heteroaryl, heteroalkylalkyl, arylalkyl, or heteroarylalkyl, in each case having the specified number of atoms and being optionally substituted as described herein. For example, "alkylamino" refers to a ^-NRxRy group, where one of ^Rx and ^Ry is ^alkyl and the other is H, and "dialkylamino" refers to a ^-NRxRy group, where both ^Rx and ^Ry are alkyl, where the alkyl group has the specified number of carbon atoms, for example, -NH- _{C1 - C4alkyl} or -N( ^C1 - _C4alkyl ). Typically, alkyl substituents on amines contain 1 to ₈ carbon atoms, preferably 1 to 6 carbon atoms, or more preferably 1 to 4 carbon atoms. The term also includes forms in which ^R and ^R, taken together with the nitrogen to which they are attached, form a 3- to 12-membered heterocyclyl or 5- to 12-membered heteroaryl ring, which itself may optionally replace the heterocyclyl or heteroaryl ring as described herein, and which may contain 1 to 3 additional heteroatoms selected from N, O and S as ring members, provided that the ring does not contain two consecutive oxygen atoms.

"Halogen" or "halo" refers to fluorine, chlorine, bromine and iodine (F, Cl, Br, I). Preferably, halogen refers to fluorine or chlorine (F or Cl).

Sometimes, "heteroform" is used herein to refer to a derivative of a group (e.g., an alkyl, aryl, or acyl group) in which at least one carbon atom in the designated carbocyclic group is replaced by a heteroatom selected from N, O, and S. Thus, heteroforms of alkyl, alkenyl, alkynyl, acyl, aryl, and arylalkyl are heteroalkyl, heteroalkenyl, heteroalkynyl, heteroacyl, heteroaryl, and heteroarylalkyl, respectively. It should be understood that generally no more than two N, O, or S atoms are attached sequentially, with the exception of oxo and N or S to form a nitro or sulfonyl group.

"Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

The terms "optionally substituted" and "substituted or unsubstituted" can be used interchangeably to indicate that the particular group being described may have no non-hydrogen substituents (i.e., unsubstituted) or may have one or more non-hydrogen atom substituents (i.e., substituted). Unless otherwise specified, the total number of substituents that may be present is equal to the number of H atoms present in the unsubstituted form of the group being described, as long as the substitution is chemically reasonable. When an optional substituent is attached via a double bond (e.g., an oxo (=O) substituent), it occupies two available valences, so the total number of other substituents that may be included is reduced by 2. In cases where optional substituents are independently selected from a list of alternatives, the selected groups may be the same or different.

In one aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof,

in:

R ¹ is selected from the group consisting of H, F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C(O)R ⁵ , C ₃ -C ₈ cycloalkyl, 3 to 12-membered heterocyclyl, and 5 to 12-membered heteroaryl, wherein each of the C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy is optionally substituted with one or more R ⁶ , and each of the C ₃ -C ₈ cycloalkyl, 3 to 12-membered heterocyclyl, or 5 to 12-membered heteroaryl is optionally substituted with one or more R ⁷ ;

R ² is H, F or C ₁ -C ₄ alkyl;

L is a bond or C ₁ -C ₄ alkylene;

R ³ is selected from the group consisting of C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, OH, CN, C(O)R ⁸ , COOR ⁹ , NR ¹⁰ R ¹¹ , OR ¹² , C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, and 5 to 12 membered heteroaryl, wherein each of the C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy is optionally substituted with one or more R ⁶ , and each of the C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl is optionally substituted with one or more R ⁷ ;

R ⁴ is H, halogen or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl is optionally substituted with one or more R ⁶ ;

R ⁵ is C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

Each R ⁶ is independently OH, F, CN or C ₁ -C ₄ alkoxy;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or a 3- to 6-membered heterocyclyl;

R ⁸ is C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

R ⁹ is H or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

R ¹⁰ and R ¹¹ are independently H or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl is optionally substituted with one or more R ¹⁴ ;

R ¹² is selected from the group consisting of C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, and 5 to 12 membered heteroaryl, wherein said C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl is each optionally substituted with one or more R ⁷ ;

each R ¹³ is independently C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

R ¹⁴ and R ¹⁵ are each independently OH, F, CN or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof has an absolute stereochemistry at the carbon atom bearing the R ¹ and R ² substituents, as shown in formula (IA) or formula (IB):

in:

R ¹ , R ² , L, R ³ , R ⁴ , X and Z have the same meanings as defined for formula (I).

All aspects and embodiments described in this application in relation to formula (I) also apply to compounds of formula (I-A) or (I-B).

In common embodiments of formula (I), R ² is H.

In embodiments of general Formula (I), R ⁴ is H, Cl, Br, F, or CH ₃ . In some such embodiments, R ⁴ is H or Cl. In some embodiments, R ⁴ is H. In other embodiments, R ⁴ is Cl. In further embodiments, R ⁴ is Cl or Br.

In the compounds of formula (I), X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy, or C ₁ -C ₄ fluoroalkoxy. In some embodiments, Z is C ₁ -C ₄ alkyl, such as CH ₃ or C ₂ H ₅ (i.e., methyl or ethyl). In some embodiments, X is C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, or C ₁ -C ₄ alkoxy. In specific embodiments, X is CH ₃ , OCH ₃ , or OCHF ₂ (i.e., methyl, methoxy, or difluoromethoxy). In other embodiments, X is CH ₃ , OCH ₃ , or OCHF ₂ , and Z is CH ₃ .

In some embodiments of Formula (I), R ¹ is H or F.

In other embodiments of Formula (I), R ¹ is C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy, each of which is optionally substituted with one or more R ^6. In some such embodiments, the alkyl or alkoxy group is substituted with at least one OH or CN group. In specific embodiments, R ¹ is CH ₃ , C ₂ H ₅ , CH ₂ OH, CH ₂ CH ₂ OH, CH(OH)CH ₃ or CH ₂ CN (i.e., methyl, ethyl, hydroxymethyl, 2-hydroxyethyl, 1-hydroxyethyl or cyanomethyl). In other specific embodiments, R ¹ is OCH ₃ (i.e., methoxy).

In other embodiments of Formula (I), ^R is C(O) ^R , wherein ^R is C-C alkyl optionally substituted with one or more R. In some such embodiments, ^R is _{C - C} alkyl optionally substituted with ^OH . In specific embodiments, ^R is CH, _CHOH , _CHCHOH , or CH( _{CH )} OH, such that ^R is C(O ₎ CH, C( _O ) _CHOH , C(O) _CHCHOH , or C(O)CH( _{CH )} OH (i.e., _acetyl , α-hydroxyacetyl, 3-hydroxypropanoyl, or 2-hydroxypropanoyl).

In other embodiments of Formula (I), R ¹ is C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl, wherein said C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl is each optionally substituted with one or more R ⁷ .

In some such embodiments, R ¹ is C ₃ -C ₈ cycloalkyl optionally substituted with one or more R ^7. In some such embodiments, R ¹ is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each of which is optionally substituted with one or more R ⁷ .

In other embodiments, ^R is a 3- to 12-membered heterocyclyl optionally substituted with one or more ^R. In some such embodiments, the 3- to 12-membered heterocyclyl is selected from the group consisting of tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, pyrrolidinyl, piperidinyl, and morpholinyl, each of which is optionally substituted with one or more ^R. In other such embodiments, the 3- to 12-membered heterocyclyl is selected from the group consisting of oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl, each of which is optionally substituted with one or more ^R. In specific embodiments, the 3- to ¹² -membered heterocyclyl is oxetanyl optionally substituted with one or more R. In some such embodiments, the oxetanyl is unsubstituted.

In other such embodiments, ^R is a 5- to 12-membered heteroaryl optionally substituted with one or more ^R. In some such embodiments, ^R is a 5- or 6-membered heteroaryl. In certain embodiments, the 5- or 6-membered heteroaryl is selected from the group consisting of pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, and triazolyl, each of which is optionally substituted with one or more ^R.

In certain embodiments, when R ¹ is C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl, each R ⁷ is independently CH ₃ , OH, F, CN, OCH ₃ , ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or 3 to 6 membered heterocyclyl, wherein each R ¹³ is CH ₃ or C ₂ H ₅ optionally substituted with OH (e.g., R ¹³ is CH ₃ , CH ₂ OH, CH ₂ CH ₂ OH, or CH(CH ₃ )OH).

In compounds of formula (I), L is a bond or C ₁ -C ₄ alkylene. In some embodiments of formula (I), L is a bond. In other embodiments of formula (I), L is C ₁ -C ₄ alkylene. In specific embodiments, L is methylene or ethylene.

In the compounds of formula (I), R ³ is selected from the group consisting of C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, OH, CN, C(O)R ⁸ , COOR ⁹ , NR ¹⁰ R ¹¹ , OR ¹² , C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl and 5 to 12 membered heteroaryl, wherein each of the C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy is optionally substituted with one or more R ⁶ , and each of the C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl or 5 to 12 membered heteroaryl is optionally substituted with one or more R ⁷ .

In some embodiments, R ³ is C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or 3 to 12 membered heterocyclyl, which are optionally substituted as described above. In some embodiments, R ³ is C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy, particularly CH ₃ or OCH ₃ (ie, methyl or methoxy).

In other embodiments, ^R is a 3- to 12-membered heterocyclyl optionally substituted with one or more ^R. In some such embodiments, the 3- to 12-membered heterocyclyl is selected from the group consisting of tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, pyrrolidinyl, piperidinyl, and morpholinyl, each of which is optionally substituted with one or more ^R. In other such embodiments, the 3- to 12-membered heterocyclyl is selected from the group consisting of oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl, each of which is optionally substituted with one or more ^R. In specific embodiments, the 3- to ¹² -membered heterocyclyl is an oxetanyl optionally substituted with one or more R. In some such embodiments, the oxetanyl is unsubstituted.

In other embodiments, L is a bond, and R ³ is C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or a 3- to 12-membered heterocyclyl, which is optionally substituted as described above. In particular embodiments, L is a bond, and R ³ is C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy, particularly CH ₃ or OCH ₃ (i.e., methyl or methoxy).

In further embodiments, L is a bond, and ^R is a 3- to 12-membered heterocyclyl optionally substituted with one or more ^R. In some such embodiments, L is a bond, and the 3- to 12-membered heterocyclyl is selected from the group consisting of tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, pyrrolidinyl, piperidinyl, and morpholinyl, each of which is optionally substituted with one or more ^R. In other such embodiments, L is a bond, and the 3- to 12-membered heterocyclyl is selected from the group consisting of oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl, each of which is optionally substituted with one or more ^R. In specific embodiments, L is a bond, and the 3- to ¹² -membered heterocyclyl is oxetanyl optionally substituted with one or more R. In some such embodiments, the oxetanyl is unsubstituted.

In other such embodiments, L is C ₁ -C ₄ alkylene, and R ³ is C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, or 3 to 12 membered heterocyclyl, optionally substituted as described above.

In other embodiments, R ³ is OH, CN, C(O)R ⁸ or COOR ⁹ , wherein R ⁸ is C ₁ -C ₄ alkyl optionally substituted with one or more R ¹⁴ , and R ⁹ is H or C ₁ -C ₄ alkyl optionally substituted with one or more R ¹⁴ .

In some such embodiments, L is a bond, and R ³ is OH, CN, C(O)R ⁸ or COOR ⁹ , wherein R ⁸ and R ⁹ are as described above.

In other embodiments, L is C ₁ -C ₄ alkylene, and R ³ is OH, CN, C(O)R ⁸ or COOR ⁹ , wherein R ⁸ and R ⁹ are as described above. In particular embodiments, L is C ₁ -C ₄ alkylene, such as methylene or ethylene, and R ³ is OH or CN.

In further embodiments, R ³ is OR ¹² , C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl, wherein said C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl are each optionally substituted with one or more R ⁷ , and wherein R ¹² is C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl, each of which are optionally substituted with one or more R ⁷ .

In some such embodiments, L is a bond, and R ³ is OR ¹² , C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl, as described above. In other such embodiments, L is C ₁ -C ₄ alkylene, and R ³ is OR ¹² , C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl, as described above.

In the compounds of formula (I), each R ⁶ is independently OH, F, CN, or C ₁ -C ₄ alkoxy. In common embodiments, at least one R ⁶ is OH or F.

In the compounds of Formula (I), each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or 3- to 6-membered heterocyclyl. In some embodiments, at least one R ⁷ is C(O)R ¹³ , wherein R ¹³ is C ₁ -C ₄ alkyl and the C ₁ -C ₄ alkyl is optionally further substituted with one or more R ¹⁵ . In some embodiments, at least one R ⁷ is SO ₂ R ¹³ , wherein R ¹³ is C ₁ -C ₄ alkyl and the C ₁ -C ₄ alkyl is optionally further substituted with one or more R ¹⁵ . In other specific embodiments, at least one R ⁷ is OH, F, or C ₁ -C ₄ alkyl (e.g., CH ₃ ).

In certain embodiments, R ¹ and/or R ³ is a 3- to 12-membered heterocyclyl substituted with one or more R ⁷ , wherein at least one R ⁷ is CHO or C(O)R ¹³ , and wherein R ¹³ is CH ₃ , CH ₂ OH or CH ₂ CN, such that R ⁷ is CHO, C(O)CH ₃ , C(O)CH ₂ OH or C(O)CH ₂ CN (i.e., formyl, acetyl, hydroxyacetyl or cyanoacetyl, respectively).

In certain embodiments, ^R1 and/or ^R3 is a 3- to 12-membered heterocyclyl substituted with one or more ^R7 , wherein at least one ^R7 is _SO2R13 , and _wherein ^R13 is ^CH3 , _CH2OH , or _CH2CN , such _that ^R7 is _{SO2CH3 , SO2CH2OH} , or _{SO2CH2CN .}

In further specific embodiments, R ¹ and/or R ³ is a 3- to 12-membered heterocyclyl substituted with one or more R ⁷ , wherein at least one R ⁷ is OH.

In the compounds of formula (I), R ⁸ is C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ .

In compounds of formula (I), R ⁹ is H or C ₁ -C ₄ alkyl, wherein each said C ₁ -C ₄ alkyl is optionally substituted with one or more R ^14. In some such embodiments, R ⁹ is H. In other such embodiments, R ⁹ is C ₁ -C ₄ alkyl optionally substituted as described above.

In the compounds of formula (I), R ¹⁰ and R ¹¹ are independently H or C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁴ .

R ¹⁴ and R ¹⁵ are each independently OH, F, CN or C ₁ -C ₄ alkoxy.

In a preferred embodiment, the present invention provides a compound of formula (I), (I-A) or (I-B), or a pharmaceutically acceptable salt thereof, wherein:

R ¹ is 3 to 12 membered heterocyclyl or 5 to 12 membered heteroaryl, wherein said 3 to 12 membered heterocyclyl or 5 to 12 membered heteroaryl is each optionally substituted with one or more R ⁷ ;

^R2 is H;

L is C ₁ -C ₄ alkylene;

^R3 is OH or CN;

^R4 is H or Cl;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or a 3- to 6-membered heterocyclyl;

each R ¹³ is independently C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

each R ¹⁵ is independently OH, F, CN, or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In another preferred embodiment, the present invention provides a compound of formula (I), (I-A) or (I-B), or a pharmaceutically acceptable salt thereof, wherein:

R ¹ is H or C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is optionally substituted with one or more R ⁶ ;

^R2 is H;

L is a bond or C ₁ -C ₄ alkylene;

R ³ is selected from the group consisting of C ₁ -C ₄ alkyl, OH, CN, C(O)R ⁸ , COOR ⁹ , NR ¹⁰ R ¹¹ , C ₃ -C ₈ cycloalkyl, 3 to 12-membered heterocyclyl, and 5 to 12-membered heteroaryl, wherein each of the C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ⁶ , and each of the C ₃ -C ₈ cycloalkyl, 3 to 12-membered heterocyclyl, or 5 to 12-membered heteroaryl groups is optionally substituted with one or more R ⁷ ;

^R4 is H or Cl;

each R ⁶ is independently OH, F, CN or C ₁ -C ₄ alkoxy;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or a 3- to 6-membered heterocyclyl;

R ⁸ is C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

R ⁹ is H or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

R ¹⁰ and R ¹¹ are independently H or C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁴ ;

each R ¹³ is independently C ₁ -C ₄ alkyl, each optionally substituted with one or more R ¹⁵ ;

R ¹⁴ and R ¹⁵ are each independently OH, F, CN or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In another preferred embodiment, the present invention provides a compound of formula (I), (I-A) or (I-B), or a pharmaceutically acceptable salt thereof, wherein:

R ¹ is C ₁ -C ₄ alkoxy, which is optionally substituted with one or more R ⁶ ;

^R2 is H;

L is a bond or C ₁ -C ₄ alkylene;

R ³ is selected from the group consisting of: C ₁ -C ₄ alkyl, OH, C(O)R ⁸ and 3-12 membered heterocyclyl, wherein each of the C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ⁶ , and each of the 3-12 membered heterocyclyl groups is optionally substituted with one or more R ⁷ ;

^R4 is H or Cl;

each R ⁶ is independently OH, F, CN or C ₁ -C ₄ alkoxy;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or a 3- to 6-membered heterocyclyl;

R ⁸ is C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

each R ¹³ is independently C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

R ¹⁴ and R ¹⁵ are each independently OH, F, CN or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In another preferred embodiment, the present invention provides a compound of formula (I), (I-A) or (I-B), or a pharmaceutically acceptable salt thereof, wherein:

R ¹ is C ₁ -C ₄ alkyl, which is optionally substituted with one or more R ⁶ ;

^R2 is H;

L is a bond or C ₁ -C ₄ alkylene;

R ³ is OR ¹² ;

^R4 is H or Cl;

each R ⁶ is independently OH, F, CN or C ₁ -C ₄ alkoxy;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or a 3- to 6-membered heterocyclyl;

R ¹² is selected from the group consisting of C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, and 5 to 12 membered heteroaryl, wherein said C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl is each optionally substituted with one or more R ⁷ ;

each R ¹³ is independently C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

each R ¹⁵ is independently OH, F, CN, or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In another preferred embodiment, the present invention provides a compound of formula (I), (I-A) or (I-B), or a pharmaceutically acceptable salt thereof, wherein:

^R1 is _C1 - _C4 alkoxy;

^R2 is H;

L is a key;

R ³ is a 3- to 12-membered heterocyclyl, which is preferably selected from the group consisting of oxetanyl, tetrahydrofuranyl and tetrahydropyranyl, each of which is optionally substituted with one or more R ⁷ ;

^R4 is H or Cl;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or a 3- to 6-membered heterocyclyl;

each R ¹³ is independently C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

each R ¹⁵ is independently OH, F, CN, or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In another preferred embodiment, the present invention provides a compound of formula (I), (I-A) or (I-B), or a pharmaceutically acceptable salt thereof, wherein:

R ¹ is a 3- to 12-membered heterocyclyl, which is preferably selected from the group consisting of oxetanyl, tetrahydrofuranyl and tetrahydropyranyl, each of which is optionally substituted with one or more R ⁷ ;

^R2 is H;

L is a key;

R ³ is C ₁ -C ₄ alkoxy, which is optionally substituted with one or more R ⁶ ;

^R4 is H or Cl;

Each R ⁶ is independently OH, F, CN or C ₁ -C ₄ alkoxy;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or a 3- to 6-membered heterocyclyl;

each R ¹³ is independently C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

each R ¹⁵ is independently OH, F, CN, or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In another aspect, the present invention provides a compound of formula (II), (II-A) or (II-B), or a pharmaceutically acceptable salt thereof,

in:

R ¹ , L, R ³ and X have the meanings defined for formula (I); and

R ⁴ is H, Cl, Br, F or CH ₃ .

The embodiments of formula (I), (I-A) and (I-B) described in this application also apply to compounds of formula (II), (II-A) and (II-B), as long as they are not contradictory.

In another aspect, the present invention provides a compound of formula (III), or a pharmaceutically acceptable salt thereof,

in:

R ¹ and R ³ are taken together to form a 3- to 12-membered heterocyclyl group optionally substituted with one or more R ⁷ ;

R ² is H, F or C ₁ -C ₄ alkyl;

R ⁴ is H, halogen or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl is optionally substituted with one or more R ⁶ ;

each R ⁶ is independently OH, F, CN or C ₁ -C ₄ alkoxy;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or a 3- to 6-membered heterocyclyl;

each R ¹³ is independently C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

each R ¹⁵ is independently OH, F, CN, or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In some embodiments of Formula (III), R ² is F or CH ₃ .

In the compound of formula (III), R ¹ and R ³ are taken together to form a 3- to 12-membered heterocyclyl, which is optionally substituted with one or more R ^7. In some such embodiments, the 3- to 12-membered heterocyclyl is selected from the group consisting of azetidinyl, pyrrolidinyl, piperidinyl, and homopiperidinyl, each of which is optionally substituted with one or more R ^7. In other such embodiments, the 3- to 12-membered heterocyclyl is selected from the group consisting of oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl, each of which is optionally substituted with one or more R ⁷ .

In the compound of formula (III), each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, CHO, C(O)R ¹³ , SO ₂ R ¹³ , or 3- to 6-membered heterocyclyl. In some embodiments, R ⁷ is CHO, C(O)R ¹³ , or SO ₂ R ¹³ , wherein each R ¹³ is independently C ₁ -C ₄ alkyl optionally substituted with one or more R ^15. In some such embodiments, R ¹³ is C ₁ -C 4 alkyl optionally substituted with one or more R ¹⁵ and each ^{R 15} _is independently OH, F, CN, or C ₁ -C ₄ alkoxy. In particular embodiments, R ¹³ is C ₁ -C ₄ alkyl optionally substituted with OH. In particular embodiments, R ¹³ is CH ₃ or CH ₂ OH, such that R ⁷ is C(O)CH ₃ , C(O)CH ₂ OH, SO ₂ CH ₃ , or SO ₂ CH ₂ OH (ie acetyl, α-hydroxyacetyl, methylsulfonyl, or α-hydroxymethylsulfonyl).

In some embodiments, R ⁴ is H, CH ₃ or Cl.

In some embodiments, Z is CH ₃ .

In some embodiments, X is CH ₃ or OCH ₃ .

In a preferred embodiment, the present invention provides a compound of formula (III), or a pharmaceutically acceptable salt thereof, wherein:

R ² is F or CH ₃ ;

R ¹ and R ³ are taken together to form a 3- to 12-membered heterocyclic group, each of which is optionally substituted by one or more R ⁷ ;

R ⁴ is H, CH ₃ or Cl;

Z is CH ₃ ; and

X is CH ₃ or OCH ₃ .

In another preferred embodiment, the present invention provides a compound of formula (III), or a pharmaceutically acceptable salt thereof, wherein:

R ² is F or CH ₃ ;

R ¹ and R ³ are taken together to form a 3- to 12-membered heterocyclyl selected from the group consisting of azetidinyl, pyrrolidinyl, piperidinyl and homopiperidinyl, each of which is optionally substituted with one or more R ⁷ ;

R ⁴ is H, CH ₃ or Cl;

R ⁷ is C(O)R ¹³ or SO ₂ R ¹³ ;

each R ¹³ is independently C ₁ -C ₄ alkyl optionally substituted with one or more R ¹⁵ ;

each R ¹⁵ is independently OH, F, CN or C ₁ -C ₄ alkoxy;

Z is CH ₃ ; and

X is CH ₃ or OCH ₃ .

In another aspect, the present invention provides a compound of formula (I'), or a pharmaceutically acceptable salt thereof,

in:

R ¹ is selected from the group consisting of H, F, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C(O)R ⁵ , C ₃ -C ₈ cycloalkyl, 3 to 12-membered heterocyclyl, and 5 to 12-membered heteroaryl, wherein each of the C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy is optionally substituted with one or more R ⁶ , and each of the C ₃ -C ₈ cycloalkyl, 3 to 12-membered heterocyclyl, or 5 to 12-membered heteroaryl is optionally substituted with one or more R ⁷ ;

R ² is H, F or C ₁ -C ₄ alkyl;

L is a bond or C ₁ -C ₄ alkylene;

R ³ is selected from the group consisting of C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, OH, CN, C(O)R ⁸ , COOR ⁹ , NR ¹⁰ R ¹¹ , OR ¹² , C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, and 5 to 12 membered heteroaryl, wherein each of the C ₁ -C ₄ alkyl or C ₁ -C ₄ alkoxy is optionally substituted with one or more R ⁶ , and each of the C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl is optionally substituted with one or more R ⁷ ;

R ⁴ is H, halogen or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl is optionally substituted with one or more R ⁶ ;

R ⁵ is C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

each R ⁶ is independently OH, F, CN or C ₁ -C ₄ alkoxy;

each R ⁷ is independently C ₁ -C ₄ alkyl, OH, F, CN, C ₁ -C ₄ alkoxy, ═O, or C(O)R ¹³ ;

R ⁸ is C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

R ⁹ is H or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl groups is optionally substituted with one or more R ¹⁴ ;

R ¹⁰ and R ¹¹ are independently H or C ₁ -C ₄ alkyl, wherein each of said C ₁ -C ₄ alkyl is optionally substituted with one or more R ¹⁴ ;

R ¹² is selected from the group consisting of C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, and 5 to 12 membered heteroaryl, wherein said C ₃ -C ₈ cycloalkyl, 3 to 12 membered heterocyclyl, or 5 to 12 membered heteroaryl is each optionally substituted with one or more R ⁷ ;

each R ¹³ is independently H or C ₁ -C ₄ alkyl, wherein said C ₁ -C ₄ alkyl is each optionally substituted with one or more R ¹⁵ ;

R ¹⁴ and R ¹⁵ are each independently OH, F, CN or C ₁ -C ₄ alkoxy; and

X and Z are independently C ₁ -C ₄ alkyl, C ₁ -C ₄ fluoroalkyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ fluoroalkoxy.

In some embodiments, the compound of formula (I') or a pharmaceutically acceptable salt thereof has an absolute stereochemistry at the carbon atom bearing the ^R1 and ^R2 substituents, as shown in formula (I-A') or formula (I-B'):

in:

R ¹ , R ² , L, R ³ , R ⁴ , X and Z have the same meanings as defined for formula (I).

The embodiments of formula (I), (I-A) and (I-B) described in this application also apply to compounds of formula (I'), (I-A') and (I-B'), as long as they are not contradictory.

In another aspect, the present invention provides a compound of formula (II'), (II-A') or (II-B'), or a pharmaceutically acceptable salt thereof,

in:

R ¹ , L, R ³ and X have the same meanings as defined for formula (I'); and

R ⁴ is H, Cl, Br, F or CH ₃ .

The embodiments of formula (I), (I-A) and (I-B) described in this application also apply to compounds of formula (II'), (II-A') and (II-B'), as long as they are not contradictory.

A "pharmaceutical composition" refers to a mixture of one or more of the compounds described herein or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof as an active ingredient, and at least one pharmaceutically acceptable carrier or excipient.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of one of the formulae described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. In some embodiments, the pharmaceutical composition comprises two or more pharmaceutically acceptable carriers and/or excipients.

In some embodiments, the pharmaceutical composition may further comprise at least one additional anticancer therapeutic agent or mitigating agent. In some such embodiments, the at least one additional drug or pharmaceutical substance is an anticancer agent as described below. In some such embodiments, the combination provides an additive, greater than additive, or synergistic anticancer effect. In some such embodiments, the one or more anticancer therapeutic agents are selected from the group consisting of an antitumor agent, an anti-angiogenic agent, a signal transduction inhibitor, and an antiproliferative agent.

In one aspect, the present invention provides a method of treating abnormal cell growth in a patient, comprising administering to the patient a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof.

In some embodiments, the methods described herein further comprise administering to the patient an amount of an anticancer therapeutic or palliative agent, said amount being effective together to treat said abnormal cell growth. In some embodiments, said one or more anticancer therapeutic agents are selected from anti-tumor agents, anti-angiogenic agents, signal transduction inhibitors, and anti-proliferative agents, said amount being effective together to treat said abnormal cell growth. In some such embodiments, said anti-tumor agent is selected from the group consisting of: mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, radiation, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxic agents, anti-hormones, and anti-androgens.

In common embodiments of the methods provided herein, the abnormal cell growth is cancer. In some embodiments, the methods provided herein result in one or more of the following effects: (1) inhibiting cancer cell proliferation; (2) inhibiting cancer cell invasion; (3) inducing cancer cell apoptosis; (4) inhibiting cancer cell metastasis; or (5) inhibiting angiogenesis.

In another aspect, the present invention provides a method for treating an EZH2-mediated disorder in a patient, comprising administering to the patient a compound of the present invention, or a pharmaceutically acceptable salt thereof, in an amount effective to treat the disorder.

Unless otherwise indicated, all references to compounds of the invention in this application include reference to salts, solvates, hydrates and complexes thereof, and reference to solvates, hydrates and complexes thereof, including polymorphs, stereoisomers and isotopically radiolabeled forms.

The compounds of the present invention may exist as pharmaceutically acceptable salts (e.g., acid addition salts or base addition salts) of a compound of one of the formulae provided herein. The term "pharmaceutically acceptable salts" as used herein refers to salts that retain the biological effectiveness and properties of the parent compound. The phrase "pharmaceutically acceptable salts" as used herein, unless otherwise indicated, includes salts of acidic or basic groups that may be present in a compound of a formula disclosed herein.

For example, basic compounds of the present invention can form a variety of salts with a variety of inorganic and organic acids. Although the salts must be pharmaceutically acceptable for administration to animals, in practice it is generally desirable to initially isolate the pharmaceutically unacceptable salt of the compound of the present invention from the reaction mixture, then simply convert the pharmaceutically unacceptable salt into the free base compound by treatment with an alkaline reagent, and then convert the free base into a pharmaceutically acceptable acid addition salt. Acid addition salts of basic compounds of the present invention can be prepared by treating the basic compound with substantially equal amounts of a selected inorganic or organic acid and an aqueous solvent or a suitable organic solvent (e.g., methanol or ethanol). After evaporation of the solvent, the desired solid salt is obtained. The desired acid salt can also be precipitated by adding a suitable inorganic or organic acid to a solution of the free base in an organic solvent.

Acids that can be used to prepare pharmaceutically acceptable acid addition salts of the basic compounds form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate [i.e., 1,1'-methylene-bis(2-hydroxy-3-naphthoic acid) salt].

Examples of salts include, but are not limited to, acetate, acrylate, benzenesulfonate, benzoate (e.g., chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, and methoxybenzoate), bicarbonate, bisulfate, bisulfite, bitartrate, borate, hydrobromide, butyne-1,4-dioate, calcium edetate, camphorsulfonate, carbonate, hydrochloride, hexanoate, octanoate, clavulanate, citrate, decanoate, dihydrochloride, dihydrogen phosphate, edetate, edisylate, estolate, esylate, ethylsuccinate, formate, fumarate, glucoheptonate, gluconate, glutamate, glycolate, p-glycolylaminophenylarsonic acid salt, heptanoate, hexyne-1,6-dioate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, gamma-phosphate, -hydroxybutyrate, iodide, isobutyrate, isosulfate, lactate, lactobionate, dodecanoate, malate, maleate, malonate, mandelate, methanesulfonate, metaphosphate, methanesulfonate, methosulfate, hydrogen phosphate, mucate, naphthalenesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, nitrate, oleate, oxalate, pamoate (enbolate), hexadecanoate, pantothenate, phenylacetate, phenylbutyrate, phenylpropionate, phthalate, phosphate/diphosphate, polygalacturonate, propanesulfonate, propionate, propiolate, pyrophosphate, pyrosulfate, salicylate, stearate, subacetate, suberate, succinate, sulfate, sulfonate, sulfite, tannate, tartrate, theoclate, toluenesulfonate, triethiodode, and valerate.

Examples of suitable salts include organic salts derived from amino acids (e.g., glycine and arginine), ammonia, primary, secondary, and tertiary amines and cyclic amines (e.g., piperidine, morpholine and piperazine), and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

The compounds of the present invention that include a basic group (eg, an amino group) can form pharmaceutically acceptable salts with various amino acids in addition to the above-mentioned acids.

Acidic compounds of the present invention are capable of forming base salts with a variety of pharmacologically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts, particularly sodium and potassium salts. These salts can be prepared by conventional techniques. Chemical bases useful as reagents for preparing the pharmaceutically acceptable basic salts of the present invention include those that form non-toxic base salts with the acidic compounds herein. These salts can be prepared by any suitable method, for example, by treating the free acid with an inorganic or organic base, such as a primary, secondary, or tertiary amine, an alkali metal hydroxide, or an alkaline earth metal hydroxide. These salts can also be prepared by treating the corresponding acidic compound with an aqueous solution containing the desired pharmacologically acceptable cation, and then evaporating the resulting solution to dryness, preferably under reduced pressure. Alternatively, these salts can be prepared by mixing the acidic compound with a lower alkanol solution of the desired alkali metal alkoxide, and then evaporating the resulting solution to dryness in the manner described above. In either case, stoichiometric amounts of reagents can be used to ensure complete reaction and maximum yield of the desired end product.

Chemical bases useful as reagents for preparing pharmaceutically acceptable base salts of acidic compounds of the present invention are chemical bases that form non-toxic base salts with the acidic compounds of the present invention. Such non-toxic base salts include, but are not limited to, base salts derived from pharmacologically acceptable cations, such as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium and magnesium), ammonium or water-soluble amine addition salts (e.g., N-methylglucamine-(meglumine)), and lower alkanolammonium base salts and other pharmaceutically acceptable organic amine base salts.

Hemisalts of acids and bases can also be formed, for example hemisulphate and hemicalcium salts.

For a review of suitable salts, see Stahl and Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (Wiley-VCH, 2002). Methods for making pharmaceutically acceptable salts are known to those skilled in the art.

The salts of the present invention can be prepared according to methods known to those skilled in the art. Pharmaceutically acceptable salts of the compounds of the present invention can be readily prepared by mixing the compounds of the present invention with a solution of the desired acid or base, as appropriate. The salts can be precipitated from the solution and collected by filtration, or recovered by evaporating the solvent. The degree of ionization of the salts can vary from fully ionized to nearly non-ionized.

Those skilled in the art will appreciate that free base forms of compounds of the present invention having basic functional groups can be converted to acid addition salts by treatment with a stoichiometric excess of an appropriate acid. Typically, acid addition salts of compounds of the present invention can be converted to the corresponding free bases by treatment with a stoichiometric excess of an appropriate base (e.g., potassium carbonate or sodium hydroxide) at temperatures between about 0°C and 100°C in the presence of an aqueous solvent. The free base form can be isolated by conventional means, such as extraction with an organic solvent. In addition, acid addition salts of compounds of the present invention can be exchanged by exploiting the differential solubility of salts, the volatility or acidity of acids, or by treatment with appropriately loaded ion exchange resins. For example, reaction of a salt of a compound of the present invention with a slight stoichiometric excess of an acid having a lower pK value than the acid component of the starting salt can effect exchange. This conversion is typically performed at a temperature between about 0°C and the boiling point of the solvent used in the step. Base addition salts generally enable similar exchange reactions through the intermediate nature of the free base form.

The compounds of the present invention can exist in both unsolvated and solvated forms. When the solvent or water is firmly bound, the complex will have a perfect stoichiometry independent of moisture. However, when the solvent or water is weakly bound (such as in channel solvates and hygroscopic compounds), the water/solvent content will depend on moisture and drying conditions. In this case, non-stoichiometry will be the norm. The term "solvate" as used herein describes a molecular complex comprising a compound of the present invention and one or more pharmaceutically acceptable solvent molecules (e.g., ethanol). When the solvent is water, the term "hydrate" is used. Pharmaceutically acceptable solvates according to the present invention include hydrates and solvates in which the solvent of crystallization may be isotopically substituted, such as _D2O , _d6 -acetone, and _d6 -DMSO.

Also within the scope of the present invention are complexes, such as clathrates, drug-host inclusion complexes, wherein, in contrast to the solvates described above, the drug and host are present in stoichiometric or non-stoichiometric form. Also within the scope of the present invention are complexes containing two or more organic and/or inorganic components of the drug (which may be present in stoichiometric or non-stoichiometric form). The resulting complexes may be ionized, partially ionized, or non-ionized. For a review of complexes, see J Pharm Sci, 64(8), 1269-1288 by Haleblian (August 1975), the entire contents of which are incorporated herein by reference.

The present invention also relates to prodrugs of compounds of the formula provided herein. Thus, when administered to the patient, certain derivatives of the compounds of the present invention that may themselves have little or no pharmacological activity may, for example, be hydrolytically cleaved and converted into compounds of the present invention. Such derivatives are referred to as "prodrugs." Further information on the use of prodrugs can be found in 'Pro-drugs as Novel Delivery Systems', Vol. 14, ACS Symposium Series (T Higuchi and W Stella) and 'Bioreversible Carriers in Drug Design', Pergamon Press, 1987 (E BRoche, ed., American Pharmaceutical Association), the entire contents of which are incorporated herein by reference. As used herein, "patient" may refer to a human or animal patient.

Prodrugs according to the invention can be prepared, for example, by replacing appropriate functional groups present in the compounds of the invention with certain groups known to those skilled in the art as "pro-moieties", see, for example, "Design of Prodrugs" by HBundgaard (Elsevier, 1985), the entire contents of which are incorporated herein by reference.

Some non-limiting examples of potential prodrugs according to the present invention include:

(i) when the compound contains a carboxylic acid function (—COOH), for example, an ester in which the hydrogen is replaced by a (C ₁ -C ₈ )alkyl group;

(ii) when the compound contains an alcohol function (—OH), for example, an ether whose hydrogen is replaced by a (C ₁ -C ₆ )alkanoyloxymethyl group; and

(iii) When the compound contains a primary or secondary amine function ( _-NH2 or -NHR, where R≠H), for example, an amide in which one or both hydrogens are replaced by a metabolically labile group (e.g., amide, carbamate, urea, phosphonate, sulfonate, etc.).

Additional examples of substituent groups according to the above examples and examples of other potential prodrug types can be found in the above references.

Finally, certain compounds of the invention may themselves act as potential prodrugs of other compounds of the invention.

Also within the scope of the present invention are metabolites of the compounds of the formulae described herein, ie, compounds formed in vivo following administration of the drug.

The compounds of the formula provided in this application may have asymmetric carbon atoms. Solid lines, solid wedges, or dashed wedges are used in this application to represent carbon-carbon bonds of the compounds of the present invention. The use of solid lines to represent bonds to asymmetric carbon atoms is intended to include all possible stereoisomers (e.g., specific enantiomers, racemic mixtures, etc.) at the asymmetric carbon atoms. The use of solid lines or dashed wedges to represent bonds to asymmetric carbon atoms is intended to include only the stereoisomers shown. The compounds of the present invention may contain more than one asymmetric carbon atom. In such compounds, the use of solid lines to represent bonds to asymmetric carbon atoms is intended to include all possible stereoisomers. For example, unless otherwise indicated, it is expected that the compounds of the present invention can exist as enantiomers and diastereomers, or as racemates and mixtures thereof. The use of solid lines to represent bonds to one or more asymmetric carbon atoms in the compounds of the present invention and the use of solid lines or dashed wedges to represent bonds to the remaining asymmetric carbon atoms in the compounds of the present invention are intended to indicate the presence of a mixture of diastereomers.

Compounds of the present invention having chiral centers may exist in the form of stereoisomers (eg, racemates, enantiomers, or diastereomers).

Stereoisomers of the compounds of the formula herein may include cis and trans isomers, optical isomers (e.g., (R) and (S) enantiomers), diastereomers, geometric isomers, rotational isomers, atropisomers, conformers, and tautomers of the compounds of the present invention (including compounds exhibiting more than one isomeric type); and mixtures thereof (e.g., racemates and diastereomeric pairs). Also included are acid addition salts or base addition salts where the counterion is optically active (e.g., d-lactic acid or l-lysine) or racemic (e.g., dl-tartaric acid or dl-arginine).

When a racemate crystallizes, two different types of crystals are possible. The first is the racemic compound (true racemate) described above, in which a homogeneous crystalline form is produced containing two enantiomers in equimolar amounts. The second is a racemic mixture or conglomerate, in which two crystalline forms are produced in equimolar amounts, each containing one enantiomer.

Compounds of the present invention can exhibit tautomerism and structural isomerism phenomena. For example, compounds of the present invention can exist in the form of several tautomeric forms (including enol and imine forms, and ketone and enamine forms) and geometric isomers and mixtures thereof. The scope of compounds of the present invention includes all these tautomeric forms. Tautomers exist as mixtures of tautomer sets in solution. In solid form, usually a tautomer is predominant. Even if a tautomer may be described, the present invention includes all tautomers of the compounds of the formula provided.

In addition, some compounds of the present invention may form atropisomers (e.g., substituted biaryls). Atropisomers are conformational isomers that occur when rotation about a single bond in a molecule is hindered or significantly slowed due to asymmetric steric interactions with other parts of the molecule and substituents on either side of the single bond. Interconversion between atropisomers is slow enough to allow separation and isolation under predetermined conditions. The energy barrier to thermal racemization can be determined by steric hindrance to free rotation of one or more bonds forming the chiral axis.

When the compounds of the present invention contain an alkenyl or alkenylene group, geometric cis/trans (or Z/E) isomers may exist. Cis/trans isomers can be separated by conventional techniques well known to those skilled in the art (e.g., chromatography or fractional crystallization).

Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor, or separation of the racemate (or the racemate of a salt or derivative) by, for example, chiral high pressure liquid chromatography (HPLC).

Alternatively, the racemate (or racemic precursor) can be reacted with an appropriate optically active compound (e.g., ethanol, or, when the compound contains an acidic or basic group, an acid or base, such as tartaric acid or 1-phenylethylamine). The resulting diastereomeric mixture can be separated by chromatography and/or fractional crystallization, and one or both diastereomers can be converted to the corresponding pure enantiomers by methods well known to those skilled in the art.

The chiral isomers of the present invention (and their chiral precursors) can be obtained in enantiomerically enriched form by chromatography (typically HPLC) on an asymmetric resin using a mobile phase consisting of a hydrocarbon (typically heptane or hexane) containing 0 to 50% (typically 2 to 20%) isopropanol and 0 to 5% alkylamine (typically 0.1% diethylamine). The eluate is concentrated to obtain an enantiomerically enriched mixture.

Stereoisomeric aggregates can be separated by conventional techniques known to those skilled in the art, see, for example, E. L. Eliel, "Stereochemistry of Organic Compounds" (Wiley, New York, 1994), the entire contents of which are incorporated herein by reference.

As used herein, "enantiomerically pure" describes a compound that exists as a single enantiomer and is expressed in enantiomeric excess (e.e.). Preferably, where the compound exists as an enantiomer, the enantiomer is present in an enantiomeric excess of greater than or equal to about 80%, more preferably, in an enantiomeric excess of greater than or equal to about 90%, more preferably, in an enantiomeric excess of greater than or equal to about 95%, more preferably, in an enantiomeric excess of greater than or equal to about 98%, and most preferably, in an enantiomeric excess of greater than or equal to about 99%. Similarly, "diastereomerically pure" as used herein describes a compound that exists as a single diastereomer and is expressed in diastereomeric excess (d.e.). Preferably, where the compound exists as a diastereoisomer, the diastereoisomer is present in a diastereomeric excess of greater than or equal to about 80%, more preferably in a diastereomeric excess of greater than or equal to about 90%, more preferably in a diastereomeric excess of greater than or equal to about 95%, more preferably in a diastereomeric excess of greater than or equal to about 98%, and most preferably in a diastereomeric excess of greater than or equal to about 99%.

The present invention also includes isotopically labeled compounds, which are identical to the compounds described in one of the formulae provided herein, but with one or more atoms replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature.

Isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described herein, substituting an appropriate isotopically labeled reagent for the unlabeled reagent used in other processes.

Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as, but not limited to, ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, and ³⁶ Cl. Certain isotopically labeled compounds of the invention, for example, those into which radioactive isotopes (e.g., ³ H and ¹⁴ C) are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated (i.e., ³ H) and carbon-14 (i.e., ¹⁴ C) isotopes are particularly preferred for their ease of preparation and detectability. In addition, substitution with heavier isotopes (e.g., deuterium ² H) may afford certain potential therapeutic advantages resulting from potentially greater metabolic stability, for example, potentially increased in vivo half-life or potentially reduced dosage requirements, and thus may be preferred in some circumstances. Isotopically labeled compounds of the present invention can generally be prepared by carrying out the steps disclosed in the following reaction schemes and/or the Examples and Preparations, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

The compounds of the present invention may be used as crystalline products or amorphous products or mixtures thereof. The compounds of the present invention may be obtained as solid plugs, powders or films by methods such as precipitation, crystallization, freeze drying, spray drying or evaporative drying. Microwave or high-frequency drying may be used for this purpose.

Treatment methods and uses

The present invention further provides methods and uses of treatment comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof, alone or in combination with one or more other therapeutic agents or palliative agents.

In one aspect, the present invention provides a method for treating abnormal cell growth in a patient, comprising administering to the patient a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides a method for treating abnormal cell growth in a patient, comprising administering to the patient an amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, in combination with an amount of an anti-neoplastic agent, said amounts being effective together to treat said abnormal cell growth. In some such embodiments, the anti-neoplastic agent is selected from the group consisting of a mitotic inhibitor, an alkylating agent, an antimetabolite, an intercalating antibiotic, a growth factor inhibitor, radiation, a cell cycle inhibitor, an enzyme, a topoisomerase inhibitor, a biological response modifier, an antibody, a cytotoxic agent, an anti-hormone, and an anti-androgen.

The compounds of the present invention include compounds of any of the formulae described herein (i.e., compounds of Formula I, I', II, II', I-A, I-A', I-B, I-B', II-A, II-A', II-B, II-B' and III), or pharmaceutically acceptable salts thereof.

In another aspect, the present invention provides a method for treating abnormal cell growth in a patient, comprising administering to the patient an amount of a compound of the present invention or a pharmaceutically acceptable salt thereof effective to treat the abnormal cell growth.

In another aspect, the present invention provides a method for inhibiting cancer cell proliferation in a patient, comprising administering to the patient an amount of a compound of the present invention or a pharmaceutically acceptable salt thereof effective in inhibiting cell proliferation.

In another aspect, the present invention provides a method for inhibiting cancer cell invasion in a patient, comprising administering to the patient an amount of a compound of the present invention or a pharmaceutically acceptable salt thereof effective in inhibiting cell invasion.

In another aspect, the present invention provides a method for inducing apoptosis in cancer cells in a patient, comprising administering to the patient an amount of a compound of the present invention or a pharmaceutically acceptable salt thereof effective in inducing apoptosis.

In another aspect, the present invention provides a method for inducing apoptosis in a patient, comprising administering a therapeutically effective amount of a compound of one of the formulae described herein or a pharmaceutically acceptable salt thereof to the patient.

In common embodiments of the methods provided herein, the abnormal cell growth is cancer, wherein the cancer is selected from the group consisting of stromal cell cancer, ductal cell cancer, liver cancer, rhabdomyosarcoma, lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, stomach cancer, colon cancer, breast cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue cancer, urethra cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvis cancer, central nervous system (CNS) cancer, primary central nervous system (CNS) lymphoma, spinal axis cancer, brainstem glioma, pituitary adenoma, or one or more combinations of the foregoing cancers.

In some embodiments, the compounds of the present invention are selective for mutant EZH2, resulting in inhibition of trimethylation of H3K27, which is associated with certain cancers. The methods and uses provided herein can be used to treat cancers, including follicular lymphoma and diffuse large B-cell lymphoma (DLBCL).

The compounds of the present invention are useful in treating cancers, e.g., tumors, such as brain, breast, cervical, colorectal, endometrial, esophageal, gastric, head and neck, hepatocellular, laryngeal, lung, oral, ovarian, prostate, testicular and thyroid cancers and sarcomas.

As used herein, the term "therapeutically effective amount" refers to an amount of the compound administered that provides some relief from one or more of the symptoms of the condition being treated. With respect to the treatment of cancer, a therapeutically effective amount refers to an amount effective to: (1) reduce the size of a tumor, (2) inhibit (i.e., slow to some extent, preferably stop) tumor metastasis, (3) inhibit (i.e., slow to some extent, preferably stop) tumor growth or tumor invasion, and/or (4) provide some relief from (or, preferably, eliminate) one or more signs or symptoms associated with cancer.

As used herein, "patient" refers to a human or animal patient. In certain preferred embodiments, the patient is a human.

The term "treating," as used herein, unless otherwise indicated, means reversing, alleviating, inhibiting the progression of, or preventing the disorder or condition, or one or more symptoms of the disorder or condition, to which the term applies. The term "treatment," as used herein, unless otherwise indicated, refers to the act of "treating" as defined above. The term "treating" also includes adjuvant and neoadjuvant treatment of patients.

The terms "abnormal cell growth" and "hyperproliferative disorder" are used interchangeably herein.

As used herein, "abnormal cell growth," unless otherwise indicated, refers to cell growth that is not governed by normal regulatory mechanisms (e.g., loss of contact inhibition). Abnormal cell growth can be benign (non-cancerous) or malignant (cancerous). This includes the following abnormal growths: (1) tumor cells (tumors) that exhibit increased EZH2 expression; (2) benign and malignant cells of other proliferative diseases in which EZH2 is overexpressed; (3) tumors that proliferate through aberrant EZH2 activation; and (4) benign and malignant cells of other proliferative diseases in which aberrant EZH2 activation occurs.

As used herein, "cancer" refers to any malignant and/or invasive growth or tumor caused by abnormal cell growth. The term "cancer" as used herein refers to solid tumors (named after the cell type that forms the tumor), blood cancers, bone marrow cancers, or cancers of the lymphatic system. Examples of solid tumors include, but are not limited to, sarcomas and epithelial cancers. Examples of blood cancers include, but are not limited to, leukemias, lymphomas, and bone marrow cancers. The term "cancer" includes, but is not limited to, primary cancers that originate in a specific part of the body, metastatic cancers that spread from the initial site to other parts of the body, recurrence of a primary cancer after remission, and second primary cancers (new primary cancers) in a person that are different from the previous cancer. The compounds of the present invention inhibit EZH2 and are therefore useful as antiproliferative agents (e.g., cancer) or antitumor agents (e.g., effective against solid tumors) in mammals, particularly humans. In particular, the compounds of the present invention are useful in preventing and treating a variety of hyperproliferative conditions in humans, such as malignant or benign abnormal cell growth.

The compounds, compositions and methods provided herein can be used to treat cancer, including but not limited to the following cancers:

Circulatory system, such as the heart (sarcomas [angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma], myxoma, rhabdomyomas, fibromas, lipomas, and teratomas), mediastinum and ribs, and other intrathoracic organs, hemangiomas, and tumor-associated vascular tissue;

Respiratory tract, such as the nasal cavity and middle ear, paranasal sinuses, larynx, trachea, bronchi, and lungs, such as small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), bronchial cancer (squamous cell carcinoma, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar carcinoma (bronchiolar), bronchial adenoma, sarcoma, lymphoma, chondrodystrophy, and mesothelioma;

Gastrointestinal system, for example, esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), gastric cancer, pancreatic cancer (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, VIP tumor), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Karposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, dysplasia, leiomyoma);

Genitourinary system, such as kidney (adenocarcinoma, Wilms tumor [nephroblastoma], lymphoma, leukemia), bladder and/or urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma);

Liver, such as liver cancer (hepatocellular carcinoma), bile duct cancer, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma, pancreatic endocrine tumors (such as pheochromocytoma, insulinoma, vipoma, islet cell tumor, and glucagonoma);

Bone, such as osteogenic sarcoma (osteosarcoma), fibrosarcoma, fibrous histiocytic carcinoma, chondrosarcoma, Ewing's sarcoma, lymphoma (reticulum cell sarcoma), multiple myeloma, giant cell chordal carcinoma, osteochronfroma (osteochondrotic exostosis), benign enchondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor;

Nervous system, such as central nervous system (CNS) cancer, primary central nervous system (CNS) lymphoma, head cancer (osteomas, hemangiomas, granulomas, xanthomas, osteitis deformans), meningeal cancer (meningeal carcinoma, meningeal sarcoma, glioma), brain cancer (astrocytoma, medulloblastoma, glioma, ependymoma, blastoma [pinealoma], glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor), spinal neurofibroma, meningioma, glioma, sarcoma);

Reproductive system, such as gynecological, uterus (endometrial cancer), cervix (cervical cancer, precancerous cervical dysplasia), ovary (ovarian cancer [serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma], granulosa cell and theca cell tumor, Sertoli-Leydig cell tumor, malignant germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonic rhabdomyosarcoma), fallopian tube (fallopian tube cancer) and other parts related to the female genitalia; placenta, penis, prostate, testicles and other parts related to the male genitalia;

Hematologic system, such as blood (myeloid leukemia [acute and chronic], acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma [malignant lymphoma];

Oral cavity, such as the lips, tongue, gums, floor of the mouth, jaw and other parts of the mouth, parotid and other parts of the salivary glands, tonsils, oropharynx, nasopharynx, piriform sinuses, hypopharynx, and other areas within the lips, mouth, and throat;

Skin, such as melanoma, cutaneous melanoma, stromal cell carcinoma, squamous cell carcinoma, Karposi's sarcoma, dysplastic nevus, lipoma, hemangioma, dermatofibroma, and keloid;

Adrenal gland: neuroblastoma; and

Other tissues, including connective tissue and soft tissue, retroperitoneum and peritoneum, eye, intraocular melanin and accessories, breast, head and/or neck, anus, thyroid, parathyroid, adrenal glands and other endocrine glands and related tissues, secondary and unidentified malignant tumors of lymph nodes, secondary malignant tumors of the respiratory and digestive systems and secondary malignant tumors of other parts of the body.

More specifically, examples of "cancer" used in the present invention include cancers selected from the group consisting of lung cancer (NSCLC and SCLC), head and neck cancer, ovarian cancer, colon cancer, rectal cancer, anal cancer, gastric cancer, breast cancer, kidney and ureter cancer, renal cell carcinoma, renal pelvis cancer, central nervous system (CNS) cancer, primary central nervous system (CNS) lymphoma, non-Hodgkin's lymphoma, spinal axis cancer, or a combination of one or more of the above cancers.

Still more specifically, examples of "cancer" used in the present invention include cancers selected from the group consisting of lung cancer (NSCLC and SCLC), breast cancer, ovarian cancer, colon cancer, rectal cancer, anal cancer, or a combination of one or more of the foregoing cancers.

In one embodiment of the invention, non-cancerous conditions include proliferative conditions such as benign hyperplasia of the skin (eg, psoriasis) and benign hyperplasia of the prostate (eg, BPH).

In another aspect, the present invention provides a method for inhibiting cell proliferation, comprising contacting cells with an amount of a compound of the present invention or a pharmaceutically acceptable salt thereof effective in inhibiting cell proliferation.

In another aspect, the present invention provides a method for inducing cell apoptosis, comprising contacting a cell with an amount of the compound described herein that is effective in inducing cell apoptosis.

"Contacting" refers to mixing a compound of the present invention or a pharmaceutically acceptable salt thereof with cells expressing EZH2 in a manner such that the compound can directly or indirectly affect EZH2 activity. Contacting can be performed in vitro (i.e., in an artificial environment, for example, without limitation, in a test tube or culture medium) or in vivo (i.e., in a living body, for example, without limitation, in a mouse, rat, or rabbit).

In some embodiments, the cell is a cell line (eg, a cancer cell line). In other embodiments, the cell is in a tissue or tumor, and the tissue or tumor can be in a patient (including a human).

Dosage form and regimen

Administration of the compounds of the present invention can be achieved by any method that enables the compound to be delivered to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous injection, subcutaneous injection, intramuscular injection, intravascular injection or infusion), local application and rectal administration.

The dosage regimen may be adjusted to provide the optimum desired response. For example, a single bolus injection may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and consistency of dosage. Dosage unit form, as used herein, refers to physically discrete units suitable as single dosages for the mammalian patient to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect, in association with the required pharmaceutical carrier. The specifications for the dosage unit forms of the present invention are dictated by and directly depend on (a) the unique characteristics of the chemotherapeutic agent and the specific therapeutic or prophylactic effect to be achieved, and (b) the limitations of the art in formulating active compounds for treatment of patient sensitivity.

Therefore, a skilled artisan will appreciate, based on the disclosure provided herein, that dosages and dosing regimens can be adjusted according to methods well known in the art of therapeutics. That is, a maximum tolerated dose can be readily determined, and an effective dose that provides a detectable therapeutic benefit in a patient can also be established, just as each agent can be administered briefly to provide a detectable therapeutic benefit in a patient. Therefore, although specific dosages and dosing regimens are exemplified in this application, these examples are in no way limiting of the dosages and dosing regimens that can be provided to a patient when practicing the present invention.

It is noteworthy that the dosage may vary with the type and severity of the condition to be alleviated, and may include a single dose or multiple doses. It should be understood that for any particular patient, a specific dosage regimen should be adjusted over time according to the patient's needs and the professional judgment of the person administering or guiding the administration of the composition, and the dosage ranges described in this application are merely exemplary and are not intended to limit the scope and practice of the compositions of the present invention. For example, the dosage may be adjusted according to pharmacokinetic or pharmacodynamic parameters, which may include clinical effects, such as toxic effects and/or experimental values. Therefore, the present invention includes dose increments for the same patient as determined by a skilled person. Determining the appropriate dosage and dosage regimen for administering a chemotherapeutic agent is well known in the relevant art, and the skilled person will understand that it is included once provided with the teachings provided in this application.

The dosage of the compound of the present invention administered depends on the severity of the patient being treated, the disease or condition, the rate of administration, the characteristics of the compound, and the discretion of the prescribing physician. However, an effective dose is from about 0.001 to about 100 mg per kilogram of body weight per day, preferably from about 1 to about 35 mg per kilogram of body weight per day, in a single dose or divided doses. For a 70 kg person, the dosage would be from about 0.05 to about 7 grams per day, preferably from about 0.1 to about 2.5 grams per day. In some cases, a dose below the lower limit of the above range may be more appropriate, while in other cases, a larger dose may be used without causing any harmful side effects, provided that the larger dose is first divided into several smaller doses administered over the course of a day.

Preparation and route of administration

As used herein, "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not cause significant irritation to organisms and does not abrogate the biological activity and properties of the active compound.

The pharmaceutically acceptable carrier may comprise any conventional pharmaceutical carrier or excipient. The choice of carrier and/or excipient will to a large extent depend on the following factors: the particular mode of administration, the effect of the excipient on solubility and stability, and the characteristics of the dosage form.

Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents (e.g., hydrates and solvates). If desired, the pharmaceutical composition may contain additional active ingredients, such as flavorings, binders, excipients, etc. Thus, for oral administration, tablets containing various excipients (e.g., citric acid) combined with various disintegrants (e.g., starch, alginic acid, and certain complex silicates) and binders (e.g., sucrose, gelatin, and gum arabic) may be used. Non-limiting examples of excipients include calcium carbonate, calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc are often used in tableting. Solid compositions of similar pattern may also be used in soft and hard gelatin capsules. Thus, non-limiting examples of materials include lactose and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration, the active compound may be combined with various sweetening or flavoring agents, coloring agents or pigments, and, as appropriate, emulsifying or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin, or combinations thereof.

For example, the pharmaceutical composition may be in the form of tablets, capsules, pills, powders, sustained-release formulations, solutions or suspensions suitable for oral administration, in the form of sterile solutions, suspensions or emulsions suitable for parenteral injection, in the form of ointments or creams suitable for topical administration, or in the form of suppositories suitable for rectal administration.

Exemplary parenteral administration forms include solutions or suspensions of the active compound in sterile aqueous solutions, such as aqueous propylene glycol or dextrose. Such dosage forms can be suitably buffered, if necessary.

The pharmaceutical compositions may be in unit dosage form suitable for single administration of precise quantities.

Pharmaceutical compositions suitable for delivering active drugs and methods for their preparation will be readily apparent to those skilled in the art. Such compositions and methods for their preparation may be found, for example, in 'Remington's Pharmaceutical Science', 19th edition (Mack Publishing Company, 1995), the entire contents of which are incorporated herein by reference.

The compounds of the present invention may be administered orally. Oral administration may include swallowing (allowing the compound to enter the digestive tract), or buccal or sublingual administration (allowing the compound to enter the bloodstream directly from the mouth).

Formulations suitable for oral administration include solid preparations such as tablets, capsules containing granules, liquids or powders, lozenges (including liquid-filled lozenges), chewable tablets, multi- and nanoparticulates, gels, solid solutions, liposomes, films (including buccal adhesive patches), ovules, sprays and liquid preparations.

Liquid preparations include suspensions, solutions, syrups, and elixirs. Liquid preparations can be used as fillers in soft or hard capsules and typically include a carrier, such as water, ethanol, polyethylene glycol, propylene glycol, methylcellulose, or a suitable oil, and one or more emulsifiers and/or suspending agents. Liquid preparations can also be prepared, for example, by reconstituting a solid from a sachet.

The compounds of the present invention may also be used in fast dissolving, fast disintegrating dosage forms. See Liang and Chen, Expert Opinion in Therapeutic Patents, 11(6), 981-986 (2001), the entire contents of which are incorporated herein by reference.

For tablet dosage forms, the active agent may comprise 1 wt % to 80 wt %, more typically 5 wt % to 60 wt % of the dosage form. In addition to the active agent, tablets typically contain a disintegrant. Examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, cross-linked polyvinyl pyrrolidone, polyvinyl pyrrolidone, methylcellulose, microcrystalline cellulose, hydroxypropyl cellulose substituted with a lower alkyl group, starch, pregelatinized starch, and sodium alginate. Typically, the disintegrant may comprise 1 wt % to 25 wt %, preferably 5 wt % to 20 wt % of the dosage form.

Binders are often used to impart cohesive properties to tablet formulations. Suitable binders include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic gums, polyvinyl pyrrolidone, pregelatinized starch, hydroxypropyl cellulose, and hydroxypropyl methylcellulose. Tablets may also contain diluents such as lactose (monohydrate, spray-dried monohydrate, anhydrous, etc.), mannitol, xylitol, glucose, sucrose, sorbitol, microcrystalline cellulose, starch, and dibasic calcium phosphate dihydrate.

The tablets may also optionally include surfactants (e.g., sodium lauryl sulfate and polysorbate 80) and glidants (e.g., silicon dioxide and talc). When present, the amount of surfactant is typically 0.2 wt% to 5 wt% of the tablet, and the amount of glidant is typically 0.2 wt% to 1 wt% of the tablet.

Tablets also typically contain a lubricant, such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and a mixture of magnesium stearate and sodium lauryl sulfate. The amount of lubricant is typically 0.25 wt% to 10 wt% of the tablet, preferably 0.5 wt% to 3 wt% of the tablet.

Other customary ingredients include antioxidants, colorings, flavorings, preservatives and taste-masking agents.

Exemplary tablets may contain up to about 80 wt% active agent, about 10 wt% to about 90 wt% binder, about 0 wt% to about 85 wt% diluent, about 2 wt% to about 10 wt% disintegrant, and about 0.25 wt% to about 10 wt% lubricant.

Tablet blends can be directly compressed or rolled into tablets. Alternatively, tablet blends or portions of the blend can be wet granulated, dry granulated, or melt-melted, melt-congealed, or extruded before tableting. The final formulation can include one or more layers and can be coated or uncoated; or encapsulated.

The formulation of tablets is discussed in detail in "Pharmaceutical Dosage Forms: Tablets, Vol. 1," by H. Lieberman and L. Lachman, Marcel Dekker, N.Y., N.Y., 1980 (ISBN 0-8247-6918-X), which is incorporated herein by reference in its entirety.

Oral solid dosage forms can be formulated for immediate release and/or modified release. Modified release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release, and programmed release.

Suitable controlled release formulations are described in U.S. Patent No. 6,106,864. Other suitable release technologies (e.g., high energy dispersion and osmotic and coated particles) can be found in Verma et al., Pharmaceutical Technology Online, 25(2), 1-14 (2001). WO 00/35298 describes the use of chewing gum to achieve controlled release. The entire contents of which are incorporated herein by reference.

Parenteral administration

The compounds of the present invention can also be administered directly into the bloodstream, muscle, or internal organs. Suitable parenteral administration methods include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, and subcutaneous administration. Suitable parenteral administration devices include syringes with needles (including microneedles), needle-free syringes, and infusion techniques.

Parenteral formulations are typically aqueous solutions which may contain excipients such as salts, hydrocarbons and buffers (preferably at pH 3 to 9), but for some applications, parenteral formulations may be more suitably formulated as sterile non-aqueous solutions or in dry form with an appropriate vehicle (e.g., sterile, pyrogenic water).

Parenteral formulations are readily prepared under sterile conditions (eg, by lyophilization) using standard pharmaceutical techniques well known to those skilled in the art.

The solubility of the compounds of the invention used in the preparation of parenteral solutions may be increased by the use of appropriate formulation techniques (eg, the incorporation of solubilizing agents).

Parenteral formulations can be formulated for immediate release and/or controlled release purposes. Controlled release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release, and programmed release. Therefore, the compounds of the present invention may be formulated into solid, semisolid, or thixotropic liquids that serve as implantable storage dosage forms that provide controlled release of the active compound. Examples of parenteral formulations include coated stents and PGLA microspheres.

The compounds of the present invention may also be topically applied to the skin or mucous membranes, i.e., applied to the skin or mucous membranes, i.e., applied to the skin or through the skin. Typical topical formulations include gels, hydrogels, lotions, solutions, creams, ointments, dusting powders, dressings, foams, films, skin patches, glutinous rice paper wrappers, implants, sponges, fibers, bandages, and microemulsions. Liposomes may also be used. Typical carriers include ethanol, water, mineral oil, petrolatum, white petrolatum, glycerol, polyethylene glycol, and propylene glycol. Penetration enhancers may be added, see, for example, Finnin and Morgan, J Pharm Sci, 88(10), 955-958 (October 1999). Other topical administration methods include administration by electroporation, iontophoresis, ultrasound osmosis, ultrasound introduction, and microneedle or needle-free injection (e.g., Powderject ^™ , Bioject ^™ , etc.). The entire contents of these references are incorporated herein by reference.

Topical formulations can be formulated for immediate release and/or modified release. Modified release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release, and programmed release.

The compounds of the invention may also be administered intranasally or by inhalation, typically in the form of a dry powder (alone, in a mixture, for example, with lactose, or as a mixed component particle, for example, with a phospholipid (e.g., lecithin)) from a dry powder inhaler, or in the form of an aerosol from a pressurized container, pump, sprayer, nebulizer (preferably one that utilizes electrohydrodynamics to produce a fine mist), or aerosolizer, with or without an appropriate propellant (e.g., 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane). For intranasal administration, the powder may include a bioadhesive agent, such as chitosan or cyclodextrin.

The pressurized container, pump, spray, nebulizer or aerosol may contain a solution or suspension of the compound of the invention containing, for example, ethanol, aqueous ethanol or a suitable alternative for dispersing, dissolving or sustaining the release of the active compound, a propellant as a solvent and optionally a surfactant, for example, sorbitan trioleate, oleic acid or oligolactic acid.

Before being used in dry powder or suspension formulations, the compound can be micronized to a size suitable for inhalation administration (typically less than 5 microns). This can be achieved by any appropriate pulverization method (e.g., spiral jet pulverization, fluidized bed jet pulverization, supercritical fluid processing to form nanoparticles), high pressure homogenization, or spray drying.

Capsules (e.g., made of gelatin or HPMC), blisters, and cartridges for use in inhalers or insufflators can be formulated containing a powder mix of a compound of the invention, a suitable powder base (e.g., lactose or starch), and a performance modifier (e.g., L-leucine, mannitol, or magnesium stearate). Lactose can be anhydrous or monohydrate, preferably the monohydrate. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose, and trehalose.

Suitable solution formulations for nebulizers utilizing electrohydrodynamics to generate a fine mist may contain 1 μg to 20 mg of the compound of the invention per actuation, and the actuation volume may vary from 1 μL to 100 μL. A typical formulation comprises the compound of the invention, propylene glycol, sterile water, ethanol, and sodium chloride. Alternative solvents that may be used in place of propylene glycol include glycerol and polyethylene glycol.

Appropriate flavoring agents (eg, methanol and levomenthol) or sweeteners (eg, saccharin or saccharin sodium) may be added to formulations of the invention for inhaled/intranasal administration.

Formulations for inhaled/intranasal administration can be formulated for immediate release and/or modified release (e.g., using polylactic-co-glycolic acid (PGLA)). Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted-, and programmed-release.

In the case of dry powder inhalers and aerosols, a metered dose valve is used to measure the dose. The device according to the present invention is usually arranged to administer a metered dose or "puff" containing the desired amount of the compound of the present invention. The total daily dose can be administered in a single dose or, more commonly, divided into several doses administered over the day.

The compounds of the invention may be administered rectally or vaginally, for example in the form of a suppository, pessary or enema. Coconut oil is a conventional suppository base, but a variety of alternatives may be used as appropriate.

Formulations for rectal/vaginal administration can be formulated for immediate and/or modified release. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted-, and programmed-release.

The compounds of the present invention may also be administered directly to the eye or ear, typically as droplets of a micronized suspension or solution in an isotonic pH-adjusted sterile saline solution. Other formulations suitable for ophthalmic and otic administration may include ointments, biodegradable (e.g., absorbable gel sponges, collagen) and non-biodegradable (e.g., silicone) implants, wafers, lenses, and particulate or porous systems (e.g., lipid vesicles or liposomes). Polymers such as cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, cellulosic polymers (e.g., hydroxypropyl methylcellulose, hydroxyethyl cellulose, or methylcellulose), or heteropolysaccharide polymers (e.g., gellan gum) may be used in combination with a preservative (e.g., benzalkonium chloride). Such formulations may also be administered by iontophoresis.

Formulations for ocular/aural administration can be formulated for immediate release and/or modified release. Modified release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release, or programmed release.

Other technologies

The compounds of the present invention may be used in combination with soluble macromolecular entities (such as cyclodextrins and suitable derivatives thereof or polymers containing polyethylene glycol) to improve their solubility, dissolution rate, taste masking, bioavailability and/or stability for any of the above modes of administration.

Drug-cyclodextrin complexes, for example, can be used in various dosage forms and routes of administration. Both contained and uncontained complexes may be employed. As an alternative to direct complexation with the drug, cyclodextrins can be used as auxiliary additives, i.e., as carriers, diluents, or solubilizers. For these purposes, α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin are most commonly used, examples of which can be found in PCT Publication Nos. WO 91/11172, WO 94/02518, and WO 98/55148, the entire contents of which are incorporated herein by reference.

dose

The dosage of the active compound administered depends on the severity of the patient, the disease or condition, the rate of administration, the characteristics of the compound, and the discretion of the prescribing physician. However, the effective dose is typically from about 0.001 to about 100 mg per kilogram of body weight per day, preferably from about 0.01 to about 35 mg per kilogram of body weight per day, in a single dose or divided dose form. For a 70 kg person, the dosage would be from about 0.07 to about 7000 mg per day, preferably from about 0.7 to about 2500 mg per day. In some cases, a dosage below the lower limit of the above range may be more appropriate, while in other cases, a larger dose may be used without causing any harmful side effects, provided that the larger dose is usually divided into several small doses and administered within a day.

Parts Set

Because it may be desirable to administer a combination of active compounds (e.g., for the purpose of treating a particular disease or condition), the present invention encompasses the convenient combination of two or more pharmaceutical compositions (at least one of which contains a compound according to the present invention) into a package suitable for co-administering the pharmaceutical composition. Thus, the present invention comprises a package comprising two or more separate pharmaceutical compositions (at least one of which contains a compound according to the present invention) and a device for retaining the pharmaceutical compositions separately, such as a container, a vial, or a foil packet. Examples of such packages are conventional blister packs used for packaging tablets, capsules, and the like.

The kits of the present invention may be particularly suitable for administering different dosage forms, e.g., oral and parenteral dosage forms, for administering separate compositions at different dosage intervals, or for titrating separate compositions against each other. To aid compliance, the kits of the present invention typically include instructions for administration and may be provided with a memory aid.

Combination therapy

As used herein, the term "combination therapy" refers to the sequential or simultaneous administration of a compound of the invention together with at least one additional pharmaceutical substance or agent (eg, an anticancer agent).

As described above, the compounds of the present invention may be used in combination with one or more additional anticancer agents as described below. When using combination therapy, one or more additional anticancer agents and the compounds of the present invention may be administered sequentially or simultaneously. In one embodiment, the additional anticancer agent is administered to a mammal (e.g., a human) prior to administering the compounds of the present invention. In another embodiment, the additional anticancer agent is administered to a mammal after administering the compounds of the present invention. In another embodiment, the additional anticancer agent is administered to a mammal (e.g., a human) while administering the compounds of the present invention.

The present invention also relates to a pharmaceutical composition for treating abnormal cell growth in mammals (including humans), which comprises a certain amount of the above-mentioned compound of the present invention (including hydrates, solvates and polymorphs of the compound or its pharmaceutically acceptable salt), combined with one or more (preferably one to three) anti-tumor agents selected from anti-angiogenesis agents and signal transduction inhibitors and a pharmaceutically acceptable carrier, wherein the amount of the active agent and the combined anti-cancer agent as a whole is effective to treat the abnormal cell growth.

In one embodiment of the invention, the anti-tumor agent used in combination with the compounds and pharmaceutical compositions of the invention described herein is an anti-angiogenic agent (e.g., an agent that stops tumors from developing new blood vessels). Examples of anti-angiogenic agents include, for example, VEGF inhibitors, VEGFR inhibitors, TIE-2 inhibitors, PDGFR inhibitors, angiopoietin inhibitors, PKCβ inhibitors, COX-2 (cyclooxygenase II) inhibitors, integrins (α-v/β-3), MMP-2 (matrix metalloproteinase 2) inhibitors, and MMP-9 (matrix metalloproteinase 9) inhibitors.

Preferred anti-angiogenic agents include sunitinib (Sutent ^™ ), bevacizumab (Avastin ^™ ), axitinib (AG 13736), SU 14813 (Pfizer), and AG13958 (Pfizer).

Additional anti-angiogenic agents include vatalanib (CGP 79787), sorafenib (Nexavar ^{™), pegaptanib octasodium (Macugen™} ), vandetanib (Zactima ^™ ), PF-0337210 (Pfizer ⁾ , SU 14843 (Pfizer), AZD 2171 (AstraZeneca), ranibizumab (Lucentis ^™ ), Neovastat ^™ (AE 941), tetrathiomolybdate (Coprexa ^™ ), AMG 706 (Amgen), VEGF Trap (AVE 0005), CEP 7055 (Sanofi-Aventis), XL880 (Exelixis), telatinib (BAY 57-9352) and CP-868,596 (Pfizer).

Other anti-angiogenic agents include enzastaurin (LY 317615), midostaurin (CGP 41251), perifosine (KRX 0401), teprenone (Selbex ^™ ), and UCN 01 (Kyowa Hakko).

Examples of other anti-angiogenic agents that can be used in combination with the compounds and pharmaceutical compositions of the invention described herein include celecoxib (Celebrex ^™ ), parecoxib (Dynastat™), deracoxib (SC 59046), lumiracoxib (Preige ^™ ), valdecoxib (Bextra ^™ ), rofecoxib (Vioxx ^™ ), iguratimod (Careram ^™ ), IP 751 ( ^Invedus ), SC-58125 (Pharmacia), and etoricoxib (Arcoxia ^™ ).

Other anti-angiogenic agents include exisulind (Aptosyn ^™ ), salsalate (Amigesic™), diflunisal (Dolobid ^™ ), ibuprofen (Motrin ^™ ), ketoprofen (Orudis ^™ ), nabumetone (Relafen™), piroxicam (Feldene ^™ ), naproxen (Aleve ^™ , Naprosyn ^™ ), diclofenac (Voltaren ^™ ), indomethacin (Indocin ^™ ), sulindac (Clinoril ^™ ), tolmetin (Tolectin ^™ ), etodolac (Lodine ^™ ), and dapoxetine (Dapoxetine ^{™ )} . ), ketorolac (Toradol ^™ ), and oxaprozin (Daypro ^™ ).

Other anti-angiogenic agents include ABT 510 (Abbott), apratastat (TMI 005), AZD8955 (AstraZeneca), incyclinide (Metastat ^™ ), and PCK 3145 (Procyon).

Other anti-angiogenic agents include acitretin (Neotigason ^™ ), plitidepsin (aplidine ^™ ), cilengtide (EMD 121974), combretastatin A4 (CA4P), fenretinide (4HPR), halofuginone (Tempostatin ^™ ), methoxyestradiol (Panzem ^™ ), PF-03446962 (Pfizer), rebimastat (BMS 275291), catumaxomab (Removab ^™ ), lenalidomide (Revlimid ^™ ), squalamine (EVIZON ^™ ), thalidomide (Thalomid ^™ ), Ukrain ^™ (NSC 631570), Vitaxin ^™ (MEDI 522), and zoledronic acid (Zometa ^™ ).

In another embodiment, the anticancer agent is a so-called signal transduction inhibitor (i.e., a mechanism that inhibits the fundamental processes of cell growth, differentiation, and survival that regulate the communication between molecules in the cell). Signal transduction inhibitors include small molecules, antibodies, and antisense molecules. Signal transduction inhibitors include, for example, kinase inhibitors (i.e., tyrosine kinase inhibitors or serine/threonine kinase inhibitors) and cell cycle inhibitors. More specifically, signal transduction inhibitors include, for example, farnesyl protein transferase inhibitors, EGF inhibitors, ErbB-1 (EGFR), ErbB-2, pan erb, IGF1R inhibitors, MEK, c-Kit inhibitors, FLT-3 inhibitors, K-Ras inhibitors, PI3 kinase inhibitors, JAK inhibitors, STAT inhibitors, Raf kinase inhibitors, Akt inhibitors, mTOR inhibitors, p70S6 kinase inhibitors, WNT pathway inhibitors, and so-called multi-targeted kinase inhibitors.

Preferred signaling inhibitors include gefitinib (Iressa ^™ ), cetuximab (Erbitux™), erlotinib (Tarceva ^™ ), trastuzumab (Herceptin ^™ ), sunitinib (Sutent ^™ ), imatinib (Gleevec ^™ ), and PD325901 (Pfizer ⁾ .

Examples of other signal transduction inhibitors that can be used in combination with the compounds and pharmaceutical compositions of the invention described herein include BMS 214662 (Bristol-Myers Squibb), lonafarnib (Sarasar ^™ ), pelitrexol (AG 2037), matuzumab (EMD 7200), nimotuzumab (TheraCIM h-R3 ^™ ), panitumumab (Vectibix ^™ ), vandetanib (Zactima ^™ ), pazopanib (SB 786034), ALT 110 (Alteris Therapeutics), BIBW 2992 (Boehringer Ingelheim), and Cervene ^™ (TP 38).

Examples of other signaling inhibitors include PF-2341066 (Pfizer), PF-299804 (Pfizer), canertinib (CI 1033), pertuzumab (Omnitarg ^™ ), lapatinib (Tycerb ^™ ), pelitinib (EKB 569), miltefosine (Miltefosin™), BMS ^{599626 (Bristol-Myers Squibb), Lapuleucel-T (Neuvenge™ )} , NeuVax ^™ (E75 cancer vaccine), Osidem ^™ (IDM 1), mubritinib (TAK-165), CP-724,714 (Pfizer), panitumumab (Vectibix ^™), ), lapatinib (Tycerb ^™ ), PF-299804 (Pfizer), pelitinib (EKB 569), and pertuzumab (Omnitarg ^™ ).

Examples of other signal transduction inhibitors include ARRY 142886 (Array Biopharm), everolimus (Certican ^™ ), zotarolimus (Endeavor ^™ ), temsirolimus (Torisel ^™ ), AP 23573 (ARIAD), and VX 680 (Vertex).

In addition, examples of other signal transduction inhibitors include XL 647 (Exelixis), sorafenib (Nexavar ^™ ), LE-AON (Georgetown University), GI-4000 (Globelmmune).

Examples of other signal transduction inhibitors include ABT 751 (Abbott), avocidib (flavopiridol), BMS 387032 (Bristol Myers), EM 1421 (Erimos), indisulam (E 7070), seliciclib (CYC 200), BIO 112 (Onc Bio), BMS 387032 (Bristol-Myers Squibb), PD 0332991 (Pfizer), and AG 024322 (Pfizer).

The present invention encompasses the use of the compounds of the present invention in conjunction with conventional anti-tumor agents, including, but not limited to, hormone modulators, such as hormones, anti-hormones, androgen agonists, androgen antagonists, and anti-estrogen therapeutics, histone deacetylase (HDAC) inhibitors, gene silencers or gene activators, ribonucleases, proteolytic agents, topoisomerase I inhibitors, camptothecin derivatives, topoisomerase II inhibitors, alkylating agents, antimetabolites, poly (ADP-ribose) polymerase-1 (PARP-1) inhibitors, tubulin inhibitors, antibiotics, plant-derived spindle formation inhibitors, platinum coordination compounds, gene therapy agents, antisense oligonucleotides, vascular targeting agents (VTAs), and statins.

Examples of conventional antineoplastic agents for use in combination therapy with the compounds of the present invention, optionally with one or more other agents, include, but are not limited to, glucocorticoids, such as dexamethasone, prednisone, prednisolone, methylprednisolone, hydrocortisone, and progestins, such as medroxyprogesterone, megestrol acetate (Megace), mifepristone (RU-486), selective estrogen receptor inhibitors (SERIs), estrogen receptor agonists (ER ... SERMs, such as tamoxifen, raloxifene, lasofoxifene, afimoxifene, arzoxifene, bazedoxifene, fispemifene, ormeloxifene, ospemifene, tesmilifene, toremifene, trilostane, CHF 4227 (Cheisi), selective estrogen receptor downregulators (SERDs), such as fulvestrant, exemestane (Aromasin), anastrozole (Arimidex), atamestane, fadrozole, letrozole (Femara), gonadotropin-releasing hormone (GnRH, commonly called progesterone-releasing hormone [LHRH]) agonists, such as buserelin (Suprefact), goserelin (Zoladex), leuprorelin (leuprorelin), and leuprorelin (leuprorelin). orelin (Lupron), triptorelin (Trelstar), abarelix (Plenaxis), bicalutamide (Casodex), cyproterone, flutamide (Eulexin), megestrol, nilutamide (Nilandron), osaterone, dutasteride, epristeride, finasteride, serenoa repens), PHL 00801, abarelix, goserelin, leuprorelin, triptorelin, bicalutamide, tamoxifen, exemestane, anastrozole, fadrozole, formestane, letrozole, and combinations thereof.

Other examples of conventional anti-tumor agents that can be used in combination with the compounds of the present invention include, but are not limited to, suberolanilide hydroxamic acid (SAHA, Merck Inc./Aton Pharmaceuticals), ester peptides (FR901228 or FK228), G2M-777, MS-275, pivaloyloxymethyl butyrate and PXD-101; onconase (ranpirnase), PS-341 (MLN-341), Velcade (bortezomib), 9-aminocamptothecin, belotecan, BN-80915 (Roche), camptothecin, diflomotecan, edotecarin, exatecan (Daiichi), gimatecan, 10-hydroxycamptothecin, irinotecan hydrochloride (irinotecan hydrochloride), and irinotecan hydrochloride. HCl) (Camptosar), lurtotecan, Orathecin (rubitecan, Supergen), SN-38, topotecan, camptothecin, 10-hydroxycamptothecin, 9-aminocamptothecin, irinotecan, SN-38, edotecarin, topotecan, aclarubicin, adriamycin, amonafide, amrubicin, annamycin , daunorubicin, doxorubicin, elsamitrucin, epirubicin, etoposide, idarubicin, galarubicin, hydroxycarbamide, nemorubicin, novantrone (mitoxantrone), pirarubicin, pixantrone, procarbazine, rebeccamycin, sobuzoxane, tafluposide, valrubicin, Zinecard (dexrazoxane), nitrogen mustard N-oxide, cyclophosphamide, AMD-473, altretamine, AP-5280, apaziquone, brostallicin, bendamustine, busulfan, carboquone, carbophosphamide carmustine, chlorambucil, dacarbazine, estramustine, fotemustine, glufosfamide, ifosfamide, KW-2170, lomustine, mafosfamide, mechlorethamine, melphalan, mitobronitol, mitolactol, mitomycin C C), mitoxatron, nimustine, ranimustine, temozolomide, thiotepa, and platinum-coordinated alkylating compounds, such as cisplatin, paraplatin (carboplatin), eptaplatin, lobaplatin, nedaplatin, eloxatin (oxaliplatin, Sanofi), streptozocin, satrplatin, and combinations thereof.

The present invention also encompasses the use of a compound of the present invention in combination with a dihydrofolate reductase inhibitor (e.g., methotrexate and NeuTrexin (trimetrexate glucuronate)), a purine antagonist (e.g., 6-mercaptopurine riboside, mercaptopurine, 6-thioguanine, cladribine, clofarabine (Clolar), fludarabine, nelarabine, and raltitrexed), a pyrimidine antagonist (e.g., 5-fluorouracil (5-FU), Alimta (pemetrexed disodium, LY231514, MTA), capecitabine (Xeloda ^™ ), cytosine arabinoside, Gemzar ^™ (gemcitabine, Eli Lilly), Tegafur (UFT), and ... Orzel or Uforal in combination with TS-1 (including Tegafur, gimestat, and otostat), doxifluridine, carmofur, cytarabine (including ocfosfate, stearyl phosphate, sustained-release, and liposomal forms), enocitabine, 5-aminoribofuranosyl triazone (Vidaza), decitabine ( ne) and ethynylcytidine), and other antimetabolites, such as eflornithine, hydroxyurea, leucovorin, nolatrexed (Thymitaq), triapine, trimetrexate, N-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamate, AG-014699 (Pfizer Inc.), ABT-472 (Abbott Laboratories), INO-1001 (Inotek Pharmaceuticals), KU-0687 (KuDOS Pharmaceuticals), GPI 18180 (Guilford Pharm Inc), and combinations thereof.

Other examples of conventional anti-tumor cytotoxic agents for use in combination therapy with the compounds of the invention and optionally one or more other agents include, but are not limited to, Abraxane (Abraxis BioScience, Inc.), Batabulin (Amgen), EPO 906 (Novartis), Vinflunine (Bristol-Myers Squibb Company), actinomycin D, bleomycin, mitomycin C, and daptomycin D. C), neocarzinostatin (Zinostatin), vinblastine, vincristine, vindesine, vinorelbine (Navelbine), docetaxel (Taxotere), ortataxel, paclitaxel (including Taxoprexin, a DHA/paclitaxel conjugate), cisplatin, carboplatin, nedaplatin, oxaliplatin (Eloxatin), Satraplatin, Camptosar, capecitabine (Xeloda), oxaliplatin (Eloxatin), Taxotere, Canfosfamide (Telcyta ^™ ), DMXAA (Antisoma), ibandronic acid ( acid), L-asparaginase, pegaspargase (Oncaspar ^™ ), efaproxiral (Efaproxyn™ -radiotherapy), bexarotene (Targretin ^™ ), Tesmilifene (DPPE - enhances cytotoxic potency), Theratope ^™ (Biomira), Tretinoin (Vesanoid ^™ ), tirapazamine (Trizaone ^™ ), motexafin gadolinium (Xcytrin ^™ ), Cotara ^™ (mAb), NBI-3001 (Protox Therapeutics), polyglutamate-paclitaxel ^{(Xyotax™ )} , and combinations thereof.

Other examples of conventional anti-tumor agents for use in combination therapy with the compounds of the invention and, optionally, one or more other agents include, but are not limited to, Advexin (ING 201), TNFerade (GeneVec, a compound that expresses TNFα in response to radiation therapy), RB94 (Baylor College of Medicine), Genasense (Oblimersen, Genta), Combretastatin A4P (CA4P), Oxi-4503, AVE-8062, ZD-6126, TZT-1027, Atorvastatin (Lipitor, Pfizer Inc.), Provastatin (Pravachol, Bristol-Myers Squibb), Lovastatin (Mevacor, Merck Inc.), Simvastatin (Zocor, Merck Inc.), Fluvastatin (Lescol, Novartis), Cerivastatin (Baycol, Bayer), Rosuvastatin (Crestor, AstraZeneca), Lovastatin, Niacin (Advicor, Kos Pharmaceuticals), Caduet, Lipitor, torcetrapib, and combinations thereof.

Another embodiment of the present invention of particular interest is directed to a method for treating breast cancer in a human in need of such treatment, comprising administering to said human an amount of a compound of the present invention in combination with one or more (preferably one to three) anti-neoplastic agents selected from the group consisting of trastuzumab, tamoxifen, docetaxel, paclitaxel, capecitabine, gemcitabine, vinorelbine, exemestane, letrozole and anastrozole.

In one embodiment, the present invention provides a method of treating colorectal cancer in a mammal (e.g., a human) in need of such treatment by administering an amount of a compound of the present invention in combination with one or more (preferably one to three) anti-tumor agents. Examples of specific anti-tumor agents include those commonly used in adjuvant chemotherapy, such as FOLFOX, a 5-fluorouracil (5-FU) or capecitabine (Xeloda), leucovorin, and oxaliplatin (Eloxatin). Additional examples of specific anti-tumor agents include those commonly used in chemotherapy for metastatic disease, such as FOLFOX or FOLFOX combined with bevacizumab (Avastin); and FOLFIRI, a combination of 5-FU or capecitabine, leucovorin, and irinotecan (Camptosar). Additional examples include 17-DMAG, ABX-EFR, AMG-706, AMT-2003, ANX-510 (CoFactor), aplidine (plitidepsin, Aplidin), Aroplatin, axitinib (AG-13736), AZD-0530, AZD-2171, BCG, bevacizumab (Avastin), BIO-117, BIO-145, BMS-184476, BMS-275183, BMS-528664, bortezomib (Velcade), C-1311 (Symade x), cantuzumab mertansine, capecitabine (Xeloda), cetuximab (Erbitux), clofarabine (Clofarex), CMD-193, Combretastatin, Cotara, CT-2106, CV-247, decitabine (Dacogen), E-7070, E-7820, edotecarin, EMD-273066, enzastaurin (LY-317615), epothilone B B) (EPO-906), erlotinib (Tarceva), flavopyridol, GCAN-101, gefitinib (Iressa), huA33, huC242-DM4, imatinib (Gleevec), indisulam, ING-1, irinotecan (CPT-11, Camptosar), ISIS 2503, ixabepilone, lapatinib (Tykerb), mapatumumab (HGS-ETR1), MBT-0206, MEDI-522 (Abregrin), mitomycin, MK-0457 (VX-680), MLN-8054, NB-1011, NGR-TNF, NV-1020, oblimersen (Genasense, G3139), OncoVex, ONYX 015 (CI-1042), oxaliplatin (Eloxatin), panitumumab (ABX-EGF, Vectibix), pelitinib (EKB-569), pemetrexed (Alimta), PD-325901, PF-0337210, PF-2341066, RAD-001 (Everolimus), RAV-12, Resveratrol, Rexin-G, S-1 (TS-1), seliciclib, SN-38 liposome, sodium stibogluconate (SSG), sorafenib (Nexavar), SU-14813, sunitinib (Sutent), temsirolimus (CCI 779), tetrathiomolybdate, thalomide, TLK-286 (Telcyta), topotecan (Hycamtin), trabectedin (Yondelis), vatalanib (PTK-787), vorinostat (SAHA, Zolinza), WX-UK1 and ZYC300, wherein the amount of the active agent and the amount of the combination antineoplastic agent are together effective to treat colorectal cancer.

Another embodiment of the present invention of particular interest is directed to a method for treating renal cell carcinoma in a human in need of such treatment, comprising administering to said human an amount of a compound of the present invention in combination with one or more (preferably one to three) anti-tumor agents selected from the group consisting of axitinib (AG 13736), capecitabine (Xeloda), interferon alpha, interleukin-2, bevacizumab (Avastin), gemcitabine (Gemzar), thalidomide, cetuximab (Erbitux), vatalanib (PTK-787), sunitinib (Sutent ^™ ), dapoxetine (Dapoxetine ... ), AG-13736, SU-11248, Tarceva, Iressa, Lapatinib, and Gleevec, wherein the amount of the active agent and the amount of the combination antineoplastic agent are together effective to treat renal cell carcinoma.

Another embodiment of the present invention of particular interest relates to a method for treating melanoma in a human in need of such treatment, comprising administering to said human an amount of a compound of the present invention in combination with one or more (preferably one to three) anti-tumor agents selected from the group consisting of interferon alpha, interleukin-2, temozolomide (Temodar), docetaxel (Taxotere), paclitaxel, dacarbazine (DTIC), carmustine (carmustine), dapoxetine (dapoxet ... stine (also known as BCNU), cisplatin, vinblastine, tamoxifen, PD-325,901, axitinib (AG13736), bevacizumab (Avastin), thalidomide, sorafanib, vatalanib (PTK-787), sunitinib (Sutent ^™ ), CpG-7909, AG-13736, Iressa, lapatinib, and Gleevec, wherein the amount of the active agent and the amount of the combination antineoplastic agent are together effective to treat melanoma.

Another embodiment of the present invention of particular interest is directed to a method for treating lung cancer in a human in need of such treatment, comprising administering to said human an amount of a compound of the present invention in combination with one or more (preferably one to three) anti-tumor agents selected from the group consisting of capecitabine (Xeloda), axitinib (AG 13736), bevacizumab (Avastin), gemcitabine (Gemzar), docetaxel (Taxotere), paclitaxel, premetrexed disodium (Alimta), Tarceva, Iressa, Vinorelbine, Irinotecan, Etoposide, Vinblastine, sunitinib (Sutent ^™ ), and nab-100 (Venezuela). ) and Paraplatin (carboplatin), wherein the amount of the active agent and the amount of the combined anti-tumor drug are together effective to treat lung cancer.

Synthesis method

The compounds of the present invention can be prepared according to the exemplary procedures provided herein and modifications thereof known to those skilled in the art. In addition, synthetic pathways for forming a variety of compounds that can be used as starting materials for preparing the compounds claimed in this application are described in International Application No. PCT/IB2013/060682, the entire contents of which are incorporated herein by reference.

These and other methods are exemplified in the preparation of the examples provided herein. Those skilled in the art will appreciate that the selection of starting materials and the specific sequence of steps, including, for example, forming the lactam ring, introducing or manipulating various substituents on the fused lactam or its precursors, and introducing the pyridone moiety, can be varied by selecting an appropriate synthetic strategy.

Synthesis examples are provided in the following Examples and Table 1. Table 2 provides EZH2 _IC50 values (μM) of exemplary compounds of the invention against WTEZH2 and mutant Y641N EZH2.

The following abbreviations are used throughout the examples: “Ac” means acetyl, “AcO” or “OAc” means acetoxy, “Ac ₂ O” means acetic anhydride, “ACN” or “MeCN” means acetonitrile, “AIBN” means azobisisobutyronitrile, “BOC”, “Boc” or “boc” means N-tert-butyloxycarbonyl, “Bn” means benzyl, “BPO” means dibenzoyl peroxide, “Bu” means butyl, “iBu” means isobutyl, “sBu” means sec-butyl, “tBu” means tert-butyl, “tBuOK” or “KOtBu” means potassium tert-butoxide, “CDI” means carbonyldiimidazole, “DCE” means 1,2-dichloroethane, “DCM” (CH ₂ Cl ₂ ) means dichloromethane, “DEAD” means diethyl azodicarboxylate, “DIAD” means diisopropyl azodicarboxylate, “DIPEA” or “DIEA” means diisopropylethylamine, “DBU” means 1,8-diazabicyclo[5.4.0]undec-7-ene, “DIBAL-H” means diisobutylaluminum hydride, “DMA” means N,N-dimethylacetamide, “DMAP” means 4-dimethylaminopyridine, “DME” means dimethoxyethane, and “DMF” means N,N-dimethylformamide. Methylformamide, "DMS" means dimethyl sulfide, "DMSO" means dimethyl sulfoxide, "dppf" means (diphenylphosphino)ferrocene, "DPPP" means 1,3-bis(diphenylphosphino)propane, "Et" means ethyl, "EtOAc" means ethyl acetate, "EtOH" means ethanol, "HATU" means 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, "HOAc" or "AcOH" means acetic acid, "i-Pr" or " ⁱ "Pr" means isopropyl, "IPA" means isopropyl alcohol, "KHMDS" means potassium hexamethyldisilazane (potassium bis(trimethylsilyl)amide), "LiHMDS" means lithium hexamethyldisilazane (lithium bis(trimethylsilyl)amide), "mCPBA" means meta-chloroperbenzoic acid, "Me" means methyl, "MeOH" means methanol, "Ms" means methanesulfonate (commonly known as "mesylate"), "MTBE" means methyl tert-butyl ether, "NBS" means N-bromosuccinimide, "NCS" means N-chlorosuccinimide, "NIS" means N-iodosuccinimide, "NMM" means N-methylmorpholine, "NMP" means 1-methyl-2-pyrrolidone, "Ph" means phenyl, and "RuPhos" means 2-dicyclohexylphosphine- "2',6'-diisopropoxybiphenyl,""Selectfluor" means chloromethyl-4-fluoro-1,4-diazonium bicyclo[2.2.2]octyl bis(tetrafluoroborate), "TEA" means triethylamine, "TFA" means trifluoroacetic acid, "Tf" means trifluoromethanesulfonate (commonly known as "triflate"), "THF" means tetrahydrofuran, "TMS" means trimethylsilyl, "TMSA" means trimethylsilyl azide, "TsCl" means toluenesulfonyl chloride (commonly known as "tosylate"), "SFC" means supercritical fluid chromatography, "TLC" means thin layer chromatography, "Rf" means specific shift, "~" means about, "rt" means room temperature, "h" means hour, "min" means minute, and "eq" means equivalent.

Preparation of synthetic intermediates

Compound D: 2-(Benzyloxy)-3-(Chloromethyl)-4,6-dimethylpyridine

_Ag₂O (146 g, 0.631 mol) was added to a solution of 2-hydroxy-4,6-dimethylpyridine-3-carbonitrile (85.0 g, 0.574 mol) and benzyl chloride (87.0 g, 0.688 mol) in toluene (800 mL). The reaction mixture was stirred at 110°C overnight. The reaction mixture was filtered and the solid was washed with dichloromethane. The filtrate was concentrated under vacuum and purified by column chromatography (petroleum ether/ethyl acetate) to give 2-(benzyloxy)-4,6-dimethylpyridine-3-carbonitrile (Cpd A, 89 g, 65%) as a white solid.

44.5 g x 2 batches: DIBAL-H (224 mL, 224 mmol, 1 M in toluene) was added dropwise to a stirred solution of 2-(benzyloxy)-4,6-dimethylpyridine-3-carbonitrile (Cpd A, 44.5 g, 187 mmol) in dichloromethane (500 mL) at 0-5°C. The reaction mixture was allowed to warm to room temperature and stirred for 3 hours. The mixture was quenched with 1N HCl (200 mL) and stirred vigorously for 30 minutes. The reaction mixture was neutralized with 4N NaOH (20 mL), and the biphasic mixture was filtered and rinsed with dichloromethane (500 mL). The aqueous layer was extracted with dichloromethane (200 mL), and the combined organic layers were dried over sodium sulfate and concentrated under vacuum. The residue was purified by column chromatography (petroleum ether/EtOAc) to give 2-(benzyloxy)-4,6-dimethylpyridine-3-carbaldehyde (CpdB, 70 g, 78%) as a yellow solid.

35 g x 2 batches: Sodium borohydride (6.60 g, 174 mmol) was added in several portions to a 0°C solution of 2-(benzyloxy)-4,6-dimethylpyridin-3-aldehyde (Cpd B, 35.0 g, 145 mmol) in methanol (1000 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under vacuum, and the residue was diluted with saturated aqueous _NaHCO₃ . After bubbling subsided, the aqueous solution was extracted with ethyl acetate (2 x 500 mL). The combined organic layers were dried over sodium sulfate, concentrated under vacuum, and purified by column chromatography (petroleum ether/ethyl acetate) to afford [2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methanol (Cpd C, 43 g, 61%) as a colorless oil.

21.5 g x 2 batches: Thionyl chloride (16.0 g, 133 mmol) was added to _a solution of [2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methanol (Cpd C, 21.5 g, 88.5 mmol) in anhydrous dichloromethane (400 mL) at -40°C under N₂. The mixture was stirred at -40°C for 30 minutes. The reaction mixture was poured into ice water (300 mL), and the pH was adjusted to 7-8 with _solid NaHCO₃. The mixture was separated, and the aqueous layer was extracted with dichloromethane (300 mL). The combined organic layers were washed with brine (300 mL), dried over sodium sulfate, and concentrated under vacuum. The residue was purified by column chromatography (100:1 petroleum ether/ethyl acetate) to afford 2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine (Cpd D, 27.5 g, 60%) as a white solid. ^1H NMR (400MHz, CDCl3) δ7.51-7.49 (d, 2H), 7.41-7.37 (t, 2H), 7.34-7.30 (t, 1H), 6.62 (s, 1H), 5.45 (s, 2H), 4.73 (s, 2H), 2.42 (s, 3H), 2.37 (s, 3H). MS: 261.9[M+H] ⁺ .

Compound L: 2-(benzyloxy)-3-(chloromethyl)-4-(difluoromethoxy)-6-methylpyridine

A solution of malononitrile (100 g, 1190 mmol) in anhydrous tetrahydrofuran (30 mL) was added dropwise to a cooled (-10°C) suspension of sodium hydride (60 wt% dispersion in mineral oil, 59.9 g, 1500 mmol) in anhydrous tetrahydrofuran (1200 mL), adding slowly enough to maintain the internal temperature below 5°C. After the addition was complete, the mixture was stirred at 0°C for 1.5 hours, followed by the addition of diketene (80.1 g, 1190 mmol) dropwise, adding slowly enough to maintain the internal temperature below 0°C. The mixture was stirred at -10°C for 1.5 hours, then neutralized with 4N hydrochloric acid and concentrated to remove volatiles. The remaining suspension in 4N hydrochloric acid (2000 mL) was stirred at reflux for 5 hours, then stirred at room temperature overnight. The resulting white precipitate was collected by suction filtration. The filter cake was washed sequentially with water (500 mL), ethanol (500 mL), and MTBE (300 mL). The solid was dried to give 4-hydroxy-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (Cpd E, 108 g, 60.3%) as a yellow powder. ¹ H NMR (400 MHz, DMSO-d6) δ 12.40 (br. s., 1H), 11.72 (br. s., 1H), 5.82 (s, 1H), 2.17 (s, 3H).

A suspension of 4-hydroxy-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (Cpd E, 91 g, 610 mmol), phosphorus oxychloride (195 g, 1270 mmol), and phosphorus pentachloride (265 g, 1270 mmol) in chloroform (1200 mL) was heated under reflux for 5 hours to yield a red, homogeneous mixture. The mixture was carefully poured into water (2000 mL) with stirring and then neutralized with 28% aqueous ammonium hydroxide solution. The resulting solid precipitate was filtered, washed sequentially with dichloromethane (400 mL) and ethanol (500 mL), and dried to yield 4-chloro-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (Cpd F, 78 g, 76%) as a yellow solid. ¹ H NMR (400MHz, DMSO-d6) δ 12.43 (br.s., 1H), 6.53 (s, 1H), 2.28 (s, 3H).

A suspension of 4-chloro-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile (Cpd F, 90 g, 530 mmol), silver(I) oxide (136 g, 587 mmol), and benzyl chloride (81.1 g, 641 mmol) in anhydrous toluene (1500 mL) was heated under reflux for 12 hours. The mixture was filtered through a pad of Celite®, and the filter cake was rinsed with dichloromethane (500 mL). The filtrate was concentrated to a residue (~100 g), which was purified by column chromatography (silica gel, petroleum ether/EtOAc = 50:1 to 30:1) to afford 2-(benzyloxy)-4-chloro-6-methylnicotinonitrile (Cpd G, 70 g, 51%) as a light yellow solid. ¹ H NMR (400MHz, CDCl3) δ7.49-7.47 (m, 2H), 7.40-7.33 (m, 3H), 6.91 (s, 1H), 5.05 (s, 2H), 2.50 (s, 3H).

Cesium acetate (156.0 g, 812 mmol) was added to a stirred solution of 2-(benzyloxy)-4-chloro-6-methylnicotinonitrile (Cpd G, 70 g, 270.58 mmol) in N,N-dimethylformamide (300 mL) at room temperature. The resulting mixture was stirred at 80°C for 40 hours. The mixture was diluted with ethyl acetate (500 mL) and washed with brine (3 x 400 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated to a residue (~50 g). The residue was purified by column chromatography (silica gel, petroleum ether/EtOAc = 10:1 to 3:1) to afford 2-(benzyloxy)-4-hydroxy-6-methylnicotinonitrile (Cpd H, 31 g, 48%) as a light yellow solid. ¹ H NMR (400MHz, DMSO-d6) δ 12.28 (br.s., 1H), 7.51-6.98 (m, 5H), 6.50 (s, 1H), 5.41 (s, 2H), 2.34 (s, 3H). MS 226.8[M+Na] ⁺ .

Potassium carbonate (34.5 g, 250 mmol) was added to a suspension of 2-(benzyloxy)-4-hydroxy-6-methylnicotinonitrile (CpdH, 20.0 g, 83 mmol) and sodium chlorodifluoroacetate (25.4 g, 166 mmol) in N,N-dimethylformamide (200 mL) at room temperature. The resulting mixture was heated to 100°C for 10 minutes. The reaction mixture was diluted with ethyl acetate (300 mL) and washed with saturated aqueous NH4Cl (3 x 400 mL) and brine (3 x 400 mL). The aqueous layer was back-extracted with ethyl acetate (400 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated to give a residue (18 g). The residue was purified by column chromatography (silica gel, petroleum ether/EtOAc = 50:1 to 20:1) to afford 2-(benzyloxy)-4-(difluoromethoxy)-6-methylnicotinonitrile (CpdI, 16.3 g, 67%) as a pale yellow solid. ¹ H NMR (400 MHz, CDCl3) δ 7.49-7.46 (m, 2H), 7.40-7.33 (m, 3H), 6.69 (t, J = 71 Hz, 1H), 6.67 (s, 1H), 5.51 (s, 2H), 2.52 (s, 3H).

Diisobutylaluminum hydride (1.0 M in toluene, 72 mL, 72 mmol) was added dropwise to a solution of 2-(benzyloxy)-4-(difluoromethoxy)-6-methylnicotinonitrile (Cpd I, 11 g, 38 mmol) in anhydrous dichloromethane (250 mL) at 0°C under nitrogen. After the addition was complete, the mixture was stirred at room temperature for 2.5 hours. The mixture was acidified to pH ~5 with 1 M hydrochloric acid. After stirring at room temperature for 2 hours, the mixture was neutralized with 4.0 M aqueous NaOH. The mixture was filtered through a pad of Celite® and the filter cake was rinsed with dichloromethane (300 mL). The filtrate was extracted with dichloromethane (2 x 500 mL). The combined organic layers were washed with brine (800 mL), dried over sodium sulfate, and concentrated to a residue (13.4 g). Purification by column chromatography (silica gel, petroleum ether/EtOAc = 30:1 to 10:1) afforded 2-(benzyloxy)-4-(difluoromethoxy)-6-methylnicotinaldehyde (Cpd J, 6 g, 50%) as a pale yellow solid. ¹ H NMR (400 MHz, CDCl3) δ 10.40 (s, 1H), 7.49-7.48 (m, 2H), 7.40-7.31 (m, 3H), 6.68 (t, J = 72 Hz, 1H), 6.62 (s, 1H), 5.53 (s, 2H), 2.50 (s, 3H).

Sodium borohydride (1.86 g, 49.16 mmol) was added in several portions to a solution of 2-(benzyloxy)-4-(difluoromethoxy)-6-methylnicotinaldehyde (Cpd J, 12 g, 41 mmol) in methanol (120 mL) at 0°C. After complete addition, the mixture was stirred at room temperature for 2 hours. The reaction was quenched with saturated aqueous NH4Cl (50 mL), then diluted with ethyl acetate (500 mL) and water (100 mL), and extracted with ethyl acetate (2 x 100 mL). The combined organic layers were washed with brine (300 mL), dried over sodium sulfate, and concentrated to a residue (~13.1 g). Purification by column chromatography (silica gel, petroleum ether:EtOAc = 6:1) afforded (2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methanol (Cpd K, 11.7 g, 97%) as a white solid. ¹ H NMR (400MHz, CDCl3) δ7.52-7.46 (m, 2H), 7.44-7.33 (m, 3H), 6.60 (t, J=73Hz, 1H), 6.55 (s, 1H), 5.46 (s, 2H), 2.46 (s, 3H).

Thionyl chloride (3.67 g, 30.9 mmol) was added dropwise to a solution of (2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methanol (Cpd K, 7.6 g, 26 mmol) in anhydrous dichloromethane (120 mL) at -20°C. The mixture was stirred at -20°C for 1 hour, then poured into water (50 mL) and neutralized with saturated aqueous _NaHCO₃ . The aqueous phase was extracted with dichloromethane (2 x 90 mL). The combined organic phases were dried over sodium sulfate, filtered, and concentrated to a residue (~6.1 g). Purification by silica gel chromatography (petroleum ether/EtOAc = 6:1) afforded the title compound, 2-(benzyloxy)-3-(chloromethyl)-4-(difluoromethoxy)-6-methylpyridine (Cpd L, 5.7 g, 71%), as a white solid. ¹ H NMR (400MHz, CDCl3) δ7.50 (d, J=7.2, 2H), 7.41-7.33 (m, 3H), 6.64 (t, J=73Hz, 1H), 6.56 (s, 1H), 5.48 (s, 2H), 4.69 (s, 2H), 2.47 (s, 3H). MS: 314[M+H] ⁺ .

Compound S: 2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dimethylpyridin-3-yl Hydroisoquinolin-1(2H)-one

A mixture of 3-chloro-2-methylbenzoic acid (100 g, 0.58 mol), N-chlorosuccinimide (90 g, 0.67 mol), palladium(II) acetate (14.7 g, 65.7 mmol), and N,N-dimethylformamide (1 L) was stirred overnight at 110°C under a nitrogen atmosphere. After cooling to room temperature, cesium carbonate (378 g, 1.16 mol) and iodoethane (317 g, 2.03 mol) were added, and stirring was continued at room temperature for 1.5 hours. The reaction mixture was poured into a mixture of water (1 L) and methyl tert-butyl ether (800 mL). The solid was collected by filtration, and the layers were separated. The aqueous layer was extracted with methyl tert-butyl ether (600 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (1.2 L), dried over sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel chromatography (eluting with 50:1 petroleum ether/ethyl acetate) to provide ethyl 3,6-dichloro-2-methylbenzoate (Cpd N, 110 g, -80% purity, 80% yield) as a yellow oil.

A solution of ethyl 3,6-dichloro-2-methylbenzoate (Cpd N, 120 g, 0.52 mol) and N-bromosuccinimide (147 g, 0.82 mol) in chloroform (1 L) was treated with azobisisobutyronitrile (25.3 g, 0.15 mol), and the mixture was refluxed overnight. After cooling to room temperature, the mixture was diluted with dichloromethane (800 mL) and washed with water (1.2 L). The aqueous layer was extracted with dichloromethane (800 mL). The combined organic extracts were washed with saturated aqueous sodium chloride solution (1.5 L), dried over sodium sulfate, and concentrated under vacuum to provide ethyl 2-(bromomethyl)-3,6-dichlorobenzoate (Cpd O, 160 g, 100% yield), which was used without further purification.

A solution of sodium cyanide (75.12 g, 1.53 mol) in water (300 mL) was added dropwise to a solution of ethyl 2-(bromomethyl)-3,6-dichlorobenzoate (Cpd O, 320 g, 1.03 mol) in dimethyl sulfoxide (2.4 L) at room temperature. The mixture was stirred at room temperature for 1.5 hours. The reaction mixture was poured into a mixture of water (4 L) and methyl tert-butyl ether (2 L), and the layers were separated. The organic layer was washed with water (2 L) and saturated aqueous sodium chloride solution (2 L), dried over sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel chromatography (eluting with 30:1 petroleum ether/ethyl acetate) to provide ethyl 3,6-dichloro-2-(cyanomethyl)benzoate (Cpd P, 150 g, ~75% purity, 47% yield) as a yellow oil.

Cobalt(II) chloride hexahydrate (166 g, 0.70 mol) was added to a room temperature solution of ethyl 3,6-dichloro-2-(cyanomethyl)benzoate (Cpd P, 90 g, 0.35 mol) in ethanol (1.5 L). The resulting reaction mixture was cooled to 0°C. Sodium borohydride (66.3 g, 1.74 mol) was added in several portions. The mixture was stirred at room temperature for 1 hour and then refluxed overnight. The resulting suspension was filtered and the filtrate was concentrated under vacuum. The solids in the filter cake were stirred in ethyl acetate (600 mL) and filtered. This step was repeated. The combined filtrates were added to the original filtrate residue, and the organic solution was washed with water (800 mL) and saturated aqueous sodium chloride solution (800 mL), dried over sodium sulfate, and concentrated under vacuum to afford 5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd Q, 29.3 g, 39% yield) as an off-white solid.

N-Bromosuccinimide (49.7 g, 0.279 mol) was added in portions to a solution of 5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd Q, 40 g, 0.186 mol) in concentrated sulfuric acid (200 mL) at 60°C. Stirring was continued at 60°C for 2 hours, followed by the addition of N-bromosuccinimide (5 g, 28 mmol). After stirring at 60°C for 1 hour, the mixture was poured into ice water (500 mL) and extracted with dichloromethane (3 x 500 mL). The combined organic extracts were washed with saturated aqueous sodium chloride (800 mL), dried over sodium sulfate, and concentrated under vacuum. The residue was stirred in ethyl acetate (40 mL) and petroleum ether (20 mL). The resulting solid was collected by filtration and dried in vacuo to give 7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd R, 41 g, 75% yield) as an off-white solid.

Under a nitrogen atmosphere, a solution of potassium tert-butoxide in tetrahydrofuran (1.0 M, 190 mL, 0.19 mol) was added dropwise to a cooled (0°C) solution of 7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd R, 47 g, 0.16 mol) in anhydrous N,N-dimethylformamide (500 mL). Stirring was continued at 0°C for 5 minutes, followed by the addition of 2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine (Cpd D, 40.2 g, 0.15 mol) all at once. After stirring at 0°C for 10 minutes, the mixture was treated with concentrated acetic acid (2 mL) and poured into methyl tert-butyl ether (600 mL). The organic solution was washed with water (800 mL) and saturated aqueous sodium chloride (800 mL), dried over sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel chromatography (eluting with 30:1 to 20:1 petroleum ether/ethyl acetate) to give 2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd S, 50 g, 64% yield) as an off-white solid. ^1H NMR (400MHz, DMSO-d6) δ8.08 (s, 1H), 7.45-7.43 (m, 2H), 7.32-7.29 (m, 3H), 6.76 (s, 1H), 5. 38 (s, 2H), 4.71 (s, 2H), 3.24 (t, J=6Hz, 2H), 2.72 (t, J=6Hz, 2H), 2.36 (s, 3H), 2.31 (s, 3H). MS: 521[M+H] ⁺ .

Compound T: 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo- Methyl 1,2,3,4-tetrahydroisoquinolin-7-yl)acetate

A mixture of 2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd S, 1.0 g, 1.9222 mmol), 1-(tert-butyldimethylsilyloxy)-1-methoxyethylene (1.09 g, 5.77 mmol), bis(tri-tert-butylphosphine)palladium(0) (98.2 mg, 0.192 mmol), lithium fluoride (299 mg, 11.5 mmol), and anhydrous N,N-dimethylformamide (18 mL) was degassed with nitrogen for 10 minutes. The mixture was heated at 100°C in a microwave reactor for 3 hours. Water (20 mL) was added to the reaction mixture, followed by extraction with ethyl acetate (2 x 40 mL). The combined organic layers were washed with brine (4 x 50 mL), dried over sodium sulfate, and concentrated under vacuum. The crude product was purified by silica gel chromatography (petroleum ether/EtOAc=3:1, Rf~0.45) to give methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (Cpd T, 600 mg, 60.8%) as a light yellow solid. ¹ HNMR (400MHz, CDCl3) δ7.45 (d, J=6.8Hz, 2H), 7.37-7.30 (m, 4H), 6.62 (s, 1H), 5.42 (s, 2H), 4.87 (s, 2 H), 3.80 (s, 2H), 3.72 (s, 3H), 3.28 (t, J=6.4Hz, 2H), 2.73 (t, J=6.4Hz, 2H), 2.42 (s, 3H), 2.32 (s, 3H). MS: 535.0[M+Na] ⁺ .

Compound U: 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo- Methyl 1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate

1,8-Diazabicyclo[5.4.0]undec-7-ene (0.22 mL, 1.47 mmol) was added to a solution of methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (500 mg, 0.974 mmol) and 4-acetylaminobenzenesulfonyl azide (281 mg, 1.17 mmol) in anhydrous acetonitrile (8 mL). The resulting reaction mixture was stirred at room temperature for 3 hours. After removal of the solvent, the resulting residue was purified by silica gel column chromatography (eluting with a gradient of 0 to 40% EtOAc/heptane) to afford methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate (Cpd U, 454 mg, 86% yield) as a foamy solid. LCMS: 511.10/512.10 (MN ₂ ). ^1H NMR (400MHz, CDCl3) δ7.64 (s, 1H), 7.44 (d, J = 6.60Hz, 2H), 7.29-7.39 (m, 3H), 6.63 (s, 1H), 5.47 (s, 2H), 4.89 (s, 2H), 3.86 (s, 3H), 3.30 (t, J=5.99Hz, 2H), 2.74-2.86 (m, 2H), 2.43 (s, 3H), 2.36 (s, 3H).

Compound W: 2-(2-((2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methyl)-5,8-difluoromethoxy Methyl chloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate

A solution of potassium tert-butoxide in tetrahydrofuran (1.0 M, 3.2 mL, 3.2 mmol) was added dropwise to a cooled (0°C) solution of 7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd R, 750 mg, 2.54 mmol) in anhydrous N,N-dimethylformamide (15 mL). The mixture was stirred at 0°C for 15 minutes, followed by the dropwise addition of a solution of 2-(benzyloxy)-3-(chloromethyl)-4-(difluoromethoxy)-6-methylpyridine (Cpd L, 798 mg, 2.54 mmol) in anhydrous N,N-dimethylformamide (5 mL). After stirring at 0°C for 30 minutes, the reaction was quenched with water (30 mL) and extracted with ethyl acetate (3 x 15 mL). The combined organic layers were washed sequentially with water (2×20 mL) and brine (20 mL), dried over sodium sulfate, filtered, concentrated, and purified by column chromatography (silica gel, petroleum ether/EtOAc=7:1) to give 2-((2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd V, 0.97 g, 67%) as a pale yellow solid.

A mixture of 2-((2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd V, 500 mg, 0.874 mmol), 1-(tert-butyldimethylsilyloxy)-1-methoxyethylene (494 mg, 2.62 mmol), bis(tri-tert-butylphosphine)palladium(0) (67 mg, 0.313 mmol), lithium fluoride (136 mg, 5.24 mmol), and anhydrous N,N-dimethylformamide (15 mL) was degassed with nitrogen for 10 minutes and then heated in a microwave reactor at 100°C for 3 hours. After cooling, the mixture was diluted with water (30 mL) and extracted with ethyl acetate (4 x 20 mL). The combined organic layers were washed with water (3 x 15 mL) and brine (15 mL), dried, and concentrated. The residue was purified by preparative TLC (silica gel, petroleum ether/EtOAc=2:1, Rf~0.35) to give methyl 2-(2-((2-(benzyloxy)-4-(difluoromethoxy)-6-methylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (Cpd W, 175 mg, 35.4%) as a white solid. ^1H NMR (400MHz, CDCl3) δ7.42-7.41 (m, 2H), 7.37 (s, 1H), 7.29-7.27 (m, 3H), 6.66 (t, J=72Hz, 1H), 6.62 (s, 1H), 5 .45 (s, 2H), 4.82 (s, 2H), 3.80 (s, 2H), 3.72 (s, 3H), 3.26 (t, J = 6.0Hz, 2H), 2.73 (t, J = 6.4Hz, 2H), 2.46 (s, 3H).

Compound Z: 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-7-bromo-8-chloro-3,4-dihydroisothiazolinone Quinolin-1(2H)-one

A solution of 7-amino-3,4-dihydroisoquinolin-1(2H)-one (1.01 g, 6.23 mmol) and N-chlorosuccinimide (832 mg, 6.23 mmol) in N,N-dimethylformamide (10 mL) was heated to 55°C for 5 hours. The mixture was poured into water and extracted with ethyl acetate (3x). The combined ethyl acetate layers were concentrated, and residual DMF was removed under high vacuum overnight. The resulting dark oil was purified on silica gel (Biotage SNAP, 50 g, gradient from 50 to 100% ethyl acetate in heptane) to provide 7-amino-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd X, 0.539 g, 44%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ )δ7.87 (br.s., 1H), 6.96 (d, J=8.19Hz, 1H), 6.87 (d, J=8.19Hz, 1H), 5.32 (s, 2H), 3.20 (dt, J=3.79, 6.17Hz, 2H), 2.69 (t, J=6.24Hz, 2H); MS 197[M+H] ⁺ .

A suspension of copper (I) bromide (1.04 g, 7.28 mmol) in acetonitrile (20 mL) was stirred at 60°C for 10 minutes. Isoamyl nitrite (0.348 mL, 2.91 mmol) was added, followed by 7-amino-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd X, 0.477 g, 2.43 mmol) added all at once. The reaction mixture was stirred at 60°C for 1 hour. After cooling to room temperature, saturated aqueous _NH4Cl and EtOAc were added, and the biphasic mixture was stirred vigorously for 20 minutes. The layers were separated, the organic layer was concentrated, and the residue was purified on silica gel (Biotage SNAP, 10 g, HP-Sil, gradient from 40 to 100% ethyl acetate in heptane) to provide 7-bromo-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd Y, 0.287 g, 45%) as a yellow solid. ¹ H NMR (400MHz, CDCl3) δ7.70 (d, J=8.07Hz, 1H), 7.03 (d, J=8.07Hz, 1H), 6.14 (br.s., 1H), 3.43-3.57 (m, 2H), 2.95 (t, J=6.36Hz, 2H); MS 260, 262[M+H] ⁺ .

Potassium tert-butoxide (1.3 mL, 1.3 mmol, 1.0 M in THF) was added to a cooled (0°C) solution of 7-bromo-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd Y, 0.287 g, 1.10 mmol) in N,N-dimethylformamide (10 mL). After 5 minutes, 2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine (Cpd D, 0.311 g, 1.19 mmol) was added all at once. The mixture was stirred for 30 minutes, then quenched with acetic acid (3 drops), diluted with MTBE, and washed with water (2x). The organic layer was concentrated and the resulting oil was purified on silica gel (Biotage SNAP, 10 g, gradient 0 to 25% ethyl acetate in heptane) to provide 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-7-bromo-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd Z, 0.387 g, 72%) as a clear gum. ^1H NMR (400MHz, CDCl3) δ7.61 (d, J=8.07Hz, 1H), 7.42-7.47 (m, 2H), 7.28-7.38 (m, 3H), 6.89 (d, J=8.07Hz, 1H) , 6.63(s,1H),5.43(s,2H),4.90(s,2H),3.22-3.29(m,2H),2.60-2.66(m,2H),2.42(s,3H),2.34(s,3H);MS 485,487[M+H] ⁺ .

Compound FF: 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5-bromo-8-chloro-7-iodo-3,4- Dihydroisoquinolin-1(2H)-one

Two batches were run in parallel under the following conditions and then combined for workup and purification: Oxalyl chloride (14.5 g, 9.97 mL, 114 mmol) and DMF (150 mg, 2.05 mmol) were added to a room temperature (15-20°C) solution of 2-(2-bromo-5-chlorophenyl)acetic acid (25.0 g, 100.2 mmol) in anhydrous THF (300 mL). Gas evolution began. The mixture was stirred at room temperature for 2 hours, until TLC indicated complete consumption of the starting acid. The mixture was cooled to 0°C, and 28 wt% aqueous ammonium hydroxide (154 mL) was added all at once, causing the internal temperature to rise to 40°C. The cooling bath was removed, and the solution was stirred vigorously at room temperature for 1 hour. The two batches were combined, diluted with water (500 mL), and extracted with ethyl acetate (2 x 1000 mL). The combined organic extracts were washed with water (2 x 500 mL), 1N hydrochloric acid (500 mL), and brine (500 mL), then dried over anhydrous sodium sulfate and concentrated to yield a crude yellow solid (~50 g). The crude product was crystallized from 5/1 petroleum ether/ethyl acetate (200 mL x 2) and dried to yield 2-(2-bromo-5-chlorophenyl)acetamide (Cpd AA, 44.0 g, 88% yield over two batches) as a white solid. ^1H NMR (400 MHz, CDCl3) δ 7.52 (d, J = 8.8 Hz, 1H), 7.37 (d, J = 2.8 Hz, 1H), 7.16 (dd, J = 2.8, 8.8 Hz, 1H), 5.67 (br s, 1H), 5.50 (br s, 1H), 3.70 (s, 2H).

Two batches were run in parallel under the following conditions and then combined for purification: Borane-THF complex (1.0 M in THF, 400 mL, 400 mmol) was added dropwise to a cooled (0°C) suspension of 2-(2-bromo-5-chlorophenyl)acetamide (CpdAA, 22.0 g, 88.5 mmol) in anhydrous THF (300 mL). The resulting clear solution was heated to 80°C for 2 hours and then cooled to 0°C. The mixture was quenched by the addition of water (45 mL) and concentrated hydrochloric acid (120 mL), resulting in copious gas evolution. Stirring was continued at 10-15°C for 16 hours, and the mixture was concentrated to remove the THF. The aqueous residue was cooled to 0°C, and 12N aqueous sodium hydroxide solution was added to raise the pH to 11. The basified solution was extracted with ethyl acetate (3 x 500 mL). The combined organic extracts were washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude yellow oil (~25 g). Two batches of ~25 g of the crude product were combined, treated with 4N HCl/MeOH (500 mL), and stirred at 10 to 15°C for 16 hours. The mixture was concentrated, and the residue was stirred in ethyl acetate (500 mL) for 30 minutes. The resulting white solid was collected by filtration, and the filter cake was washed with ethyl acetate (3 x 100 mL). The solid was dissolved in water (500 mL), filtered to remove insoluble material, and the filtrate was extracted with ethyl acetate (2 x 500 mL). The aqueous layer was basified to pH 10 with solid NaOH and then extracted with ethyl acetate (2 x 500 mL). The combined organic extracts were washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford 2-(2-bromo-5-chlorophenyl)ethan-1-amine (Cpd BB, 30.0 g, 72% yield over two batches) as a colorless oil. ¹ H NMR (400 MHz, CDCl 3 ) δ 7.47 (d, J = 8.4 Hz, 1H), 7.23 (d, J = 2.4 Hz, 1H), 7.07 (dd, J = 2.4, 8.4 Hz, 1H), 2.98 (t, J = 6.8 Hz, 2H), 2.86 (t, J = 6.8 Hz, 2H), 1.28 (m, 2H).

4-Nitrophenyl chloroformate (25.5 g, 127 mmol) was added to a cooled (0°C) suspension of 2-(2-bromo-5-chlorophenyl)ethan-1-amine (Cpd BB, 28.0 g, 119 mmol) and sodium carbonate (32.3 g, 304 mmol) in anhydrous 1,2-dichloroethane (600 mL). The mixture was stirred at 0°C for 30 minutes, then at 10-15°C for 16 hours. The solution was diluted with water (1000 mL) and extracted with dichloromethane (3 x 1000 mL). The combined organic extracts were washed with water (1000 mL) and brine (1000 mL), dried over anhydrous sodium sulfate, and concentrated. The crude yellow solid (~55 g) was crystallized from 5/1 petroleum ether/EtOAc (100 mL x 2) to give 4-nitrophenyl (2-bromo-5-chlorophenethyl)carbamate (Cpd CC, 40.0 g, 84% yield) as a white solid. ¹ H NMR (400 MHz, CDCl3) δ 8.25 (d, J = 9.2 Hz, 2H), 7.51 (d, J = 8.4 Hz, 1H), 7.31 (m, 3H), 7.13 (dd, J = 2.0, 8.4 Hz, 1H), 5.22 (br s, 1H), 3.57 (t, J = 6.8 Hz, 2H), 3.05 (t, J = 6.8 Hz, 2H).

Trifluoromethanesulfonic acid (150 g, 1000 mmol) was added dropwise to a cooled (0°C) suspension of 4-nitrophenyl (2-bromo-5-chlorophenethyl)carbamate (Cpd CC, 40.0 g, 100 mmol) in anhydrous 1,2-dichloroethane (300 mL). The solid gradually dissolved during the addition, yielding a clear yellow solution. The mixture was stirred at 0°C for 10 minutes, then heated at 60-70°C for 3 hours. The resulting brown solution was poured into ice water (1000 mL) and stirred until the ice had completely melted. The layers were separated, and the aqueous layer was extracted with dichloromethane (2 x 1000 mL). The combined organic layers were washed with 2N aqueous sodium hydroxide (3 x 500 mL), water (500 mL), and brine (500 mL), then dried over anhydrous sodium sulfate and concentrated. The crude brown solid (~30 g) was crystallized from 2/1 petroleum ether/ethyl acetate (150 mL x 2) to give 5-bromo-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (CpdDD, 20.7 g, 80% yield) as a brown solid. ¹ H NMR (400 MHz, DMSO-d6) δ 8.25 (br s, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.35 (d, J = 8.4 Hz, 1H), 3.12 (t, J = 4.4 Hz, 2H), 2.95 (t, J = 6.2 Hz, 2H).

N-Iodosuccinimide (53.7 g, 239 mmol) was added to a cooled (0°C) solution of 5-bromo-8-chloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd DD, 20.7 g, 79.6 mmol) in 98% w/w concentrated sulfuric acid (300 mL). The resulting brown suspension was stirred at 10-15°C for 16 hours, then poured into ice-water (1000 mL) and stirred until the ice had completely melted. The resulting aqueous suspension was extracted with ethyl acetate (3 x 1000 mL). The combined organic extracts were washed with saturated aqueous _NaHSO₃ (2 x 500 mL), 2N aqueous sodium hydroxide (2 x 500 mL), and brine (500 mL), then dried over anhydrous sodium sulfate and concentrated. The crude yellow solid (~30 g) was crystallized from 1/1 petroleum ether/ethyl acetate (100 mL x 2) to afford 5-bromo-8-chloro-7-iodo-3,4-dihydroisoquinolin-1(2H)-one (Cpd EE, 23.0 g, 75% yield) as an off-white solid. ¹ H NMR (400 MHz, DMSO-d6) δ ppm 8.35 (br. s, 1H), 8.33 (s, 1H), 3.30-3.25 (2H, m), 2.89 (t, J = 6.0 Hz, 2H). MS: 386 [M+H] ⁺ .

Potassium tert-butoxide (1.0 M solution in THF, 7.30 mL, 7.30 mmol) was added dropwise to a cooled (0°C) suspension of 5-bromo-8-chloro-7-iodo-3,4-dihydroisoquinolin-1(2H)-one (Cpd EE, 2.35 g, 6.08 mmol) in anhydrous DMF (30 mL). The mixture was stirred at 0°C for 30 minutes, followed by the addition of a solution of 2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine (Cpd D, 1.75 g, 6.69 mmol) in anhydrous DMF (10 mL). Stirring was continued at 0°C for 30 minutes. The reaction mixture was extracted by partitioning between ethyl acetate (100 mL) and water (100 mL). The organic phase was washed with water (1 x 100 mL) and brine (1 x 100 mL), dried over sodium sulfate, concentrated to dryness, and purified by silica gel chromatography (eluting with a gradient of 0 to 40% ethyl acetate in heptane) to provide 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5-bromo-8-chloro-7-iodo-3,4-dihydroisoquinolin-1(2H)-one (Cpd FF, 2.95 g, 79% yield) as a gel. ¹ H NMR (400MHz, CDCl3) δppm 8.11(s, 1H), 7.40-7.47(m, 2H), 7.27-7.37(m, 3H), 6.62(s, 1H), 5.42(s, 2H), 4.85 (s, 2H), 3.25 (t, J=6.24Hz, 2H), 2.68 (t, J=6.24Hz, 2H), 2.41 (s, 3H), 2.32 (s, 3H). MS: 611,613[M+H] ⁺ .

Compound KK: 2-(Benzyloxy)-3-(Chloromethyl)-4-methoxy-6-methylpyridine

Benzyl bromide (19.1 g, 112 mmol) was added to a room temperature solution of ethyl 2,4-dihydroxy-6-methylnicotinate (20.0 g, 101.4 mmol) and silver carbonate (15.4 g, 55.8 mmol) in THF (100 mL), and the mixture was heated to 60° C. for 18 hours. After cooling to room temperature, the suspension was filtered through a pad of Celite®, and the filtrate was concentrated and purified by silica gel chromatography (eluting with 5% ethyl acetate in heptane) to give ethyl 2-(benzyloxy)-4-hydroxy-6-methylnicotinate (CpdGG, 18 g, 62% yield) as a white solid. ¹ H NMR (400MHz, DMSO-d6) δ11.14 (s, 1H), 7.40-7.44 (m, 2H), 7.36 (t, J=7.34Hz, 2H), 7.27-7.33 ( m, 1H), 6.44 (s, 1H), 5.35 (s, 2H), 4.23 (q, J=7.13Hz, 2H), 2.30 (s, 3H), 1.22 (t, J=7.09Hz, 3H). MS：288[M+H] ⁺

A solution of ethyl 2-(benzyloxy)-4-hydroxy-6-methylnicotinate (Cpd GG, 18.0 g, 62.6 mmol) and potassium carbonate (9.52 g, 68.9 mmol) in DMF (50 mL) was stirred at room temperature for 10 minutes. Methyl iodide (9.98 g, 68.9 mmol) was then added and stirring continued at room temperature for 18 hours. The mixture was partitioned between water and ethyl acetate. The organic extract was washed with saturated aqueous NaCl, dried over sodium sulfate, and concentrated. The residue was purified by silica gel chromatography (eluting with 0 to 35% ethyl acetate in heptane) to provide ethyl 2-(benzyloxy)-4-methoxy-6-methylnicotinate (Cpd HH, 16.7 g, 89% yield) as a colorless oil. ¹ HNMR (400MHz, DMSO-d6) δ7.33-7.42 (m, 4H), 7.26-7.33 (m, 1H), 6.75 (s, 1H), 5.36 (s, 2H), 4.22 (q, J=7.13Hz, 2H), 3.83 (s, 3H), 2.39 (s, 3H), 1.20 (t, J=7.09Hz, 3H). ). MS: 302[M+H] ⁺ .

A 2.0 M solution of lithium aluminum hydride in THF was added dropwise to a cooled (0°C) solution of ethyl 2-(benzyloxy)-4-methoxy-6-methylnicotinate (Cpd HH, 16.7 g, 55.4 mmol) in THF (100 mL). After the addition was complete, the solution was allowed to gradually warm to room temperature while stirring for 18 hours. The mixture was diluted with THF (200 mL), cooled to 0°C, and quenched by the dropwise addition of water (3.4 mL), 15% aqueous sodium hydroxide solution, and water (10.2 mL). The resulting slurry was stirred at room temperature for 2 hours and then filtered through a pad of Celite®. The filtrate was concentrated to provide (2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methanol (Cpd JJ, 14 g, 97% yield) as a colorless oil. ¹ H NMR (400MHz, DMSO-d6) δ7.46 (d, J=7.09Hz, 2H), 7.36 (t, J=7.40Hz, 2H), 7.25-7.3 3(m, 1H), 6.63(s, 1H), 5.35(s, 2H), 4.37-4.46(m, 3H), 3.82(s, 3H), 2.35(s, 3H). MS: 260[M+H] ⁺ .

Thionyl chloride (6.57 g, 54.7 mmol) was added dropwise to a cooled (0°C) solution of (2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methanol (Cpd JJ, 13.5 g, 52.1 mmol) in ethyl acetate (300 mL), forming a solid precipitate. The slurry was stirred in the cooling bath for 30 minutes, followed by the addition of water to dissolve the solid. The phases were separated, and the organic layer was washed with saturated aqueous NaCl, dried over sodium sulfate, and concentrated to dryness. The residue was dissolved in heptane and concentrated to dryness to provide 2-(benzyloxy)-3-(chloromethyl)-4-methoxy-6-methylpyridine (Cpd KK, 13.9 g, 95% yield) as a white solid. ¹ H NMR (400MHz, DMSO-d6) δ7.47 (d, J=7.34Hz, 2H), 7.38 (t, J=7.40Hz, 2H), 7.27- 7.34 (m, 1H), 6.71 (s, 1H), 5.40 (s, 2H), 4.66 (s, 2H), 3.89 (s, 3H), 2.38 (s, 3H). MS: 260[M+H] ⁺ .

Compound RR: 8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one

Borane-dimethyl sulfide complex (28.0 g, 35.0 mL, 369 mmol) was added dropwise over 1 hour to a cooled (0°C) solution of 5-chloro-2-methylbenzoic acid (20.0 g, 117 mmol) in anhydrous 2-methyltetrahydrofuran (200 mL). The addition rate was slow enough to maintain the internal temperature below 10°C. Gas evolution and some precipitate formation were observed. After the addition was complete, the cooling bath was removed and stirring was continued at room temperature overnight. Methanol (50 mL) was cautiously added to quench the reaction. The solution was concentrated to dryness, and the residue was partitioned between diethyl ether (200 mL) and saturated aqueous sodium bicarbonate. The organic layer was washed with brine (200 mL), dried over sodium sulfate, filtered, and concentrated to afford ethyl 2-(benzyloxy)-4-hydroxy-6-methylnicotinate (Cpd LL, 18.4 g, 100% yield) as an oil. 1HNMR (400MHz, CDCl3) δ7.36 (d, J=2.08Hz, 1H), 7.13-7.19 (m, 1H), 7.04-7.11 (m, 1H), 4.63 (s, 2H), 2.27 (s, 3H), 2.12 (s, 1H).

A solution of ethyl 2-(benzyloxy)-4-hydroxy-6-methylnicotinate (Cpd LL, 18.0 g, 115 mmol) in anhydrous toluene (300 mL) was cooled to an internal temperature below 10°C. Thionyl chloride (21.3 g, 179 mmol) was added dropwise, slowly enough to maintain the internal temperature below 10°C. The mixture was stirred at this temperature for 30 minutes, after which the cooling bath was removed and stirring continued at room temperature for 5 hours. The solution was concentrated to remove volatiles, and the residue was partitioned between ethyl acetate (200 mL) and sodium bicarbonate (200 mL). The organic phase was washed with brine (200 mL), dried over sodium sulfate, and concentrated to provide 4-chloro-2-(chloromethyl)-1-toluene (Cpd MM, 17.5 g, 87% yield) as an oil. ¹ H NMR (400MHz, CDCl3) δ7.33 (d, J=2.20Hz, 1H), 7.20-7.25 (m, 1H), 7.14 (d, J=8.07Hz, 1H), 4.55 (s, 2H), 2.39 (s, 3H).

Solid sodium cyanide (5.88 g, 120 mmol) was added all at once to a solution of 4-chloro-2-(chloromethyl)-1-toluene (Cpd MM, 17.5 g, 100 mmol) in DMSO (200.0 mL) and water (50.0 mL). The reaction was slightly exothermic and the internal temperature of the reaction mixture rose to 43°C. Stirring was continued for 1 hour. The reaction mixture was partitioned and extracted with ethyl acetate (300 mL) and water (300 mL). The organic phase was washed with sodium bicarbonate (300 mL) and brine (300 mL), dried over sodium sulfate, and concentrated to give 2-(5-chloro-2-methylphenyl)acetonitrile (Cpd NN, 16.1 g, 97% yield) as an oil. ¹ H NMR (400MHz, CDCl3) δ7.37 (d, J = 1.96Hz, 1H), 7.21-7.26 (m, 1H), 7.13-7.17 (m, 1H), 3.64 (s, 2H), 2.32 (s, 3H).

Borane-dimethyl sulfide complex (22.3 g, 293 mmol, 26.0 mL) was added dropwise to a solution of 2-(5-chloro-2-methylphenyl)acetonitrile (Cpd NN, 16.0 g, 96.6 mmol) in 2-methyltetrahydrofuran (150 mL), resulting in gas evolution. After the addition was complete, the mixture was heated under reflux for 5 hours. After cooling to room temperature, the reaction was quenched by the addition of methanol until bubbling ceased. The solution was concentrated to dryness. The residue was dissolved in methanol and treated with 4M HCl/dioxane solution (100 mL) to decompose the boron complex. The solution was concentrated to dryness. The white solid residue was dissolved in a minimum amount of methanol (~20 mL), ethyl acetate (~200 mL) was added, and the mixture was stirred vigorously until a thick paste formed. The solid was collected by filtration, washed with ethyl acetate, and dried to yield 2-(5-chloro-2-methylphenyl)ethan-1-amine hydrochloride (Cpd OO, 9.5 g, 48% yield) as a white solid. ¹ H NMR (400MHz, DMSO-d6) δ 8.24 (br.s., 3H), 7.26 (s, 1H), 7.20 (d, J = 1.34Hz, 2H), 2.84-3.02 (m, 4H), 2.27 (s, 3H). MS: 170[M+H] ⁺ .

A cooled (0°C) solution of 2-(5-chloro-2-methylphenyl)ethylamine hydrochloride (Cpd OO, 8.41 g, 40.8 mmol) in DMF (200 mL) was stirred with triethylamine (10.3 g, 102 mmol) for 10 minutes, followed by the addition of 4-nitrophenyl chloroformate (8.14 g, 38.8 mmol) as a solid all at once. A thick paste formed, and the reaction mixture turned light yellow. Stirring was continued at 0°C for 1 hour. The reaction mixture was partitioned and extracted with ethyl acetate (300 mL) and water (300 mL). The organic phase was washed with water (200 mL), 10% sodium carbonate (200 mL), and brine (200 mL), then dried over sodium sulfate and concentrated to yield 4-nitrophenyl (5-chloro-2-methylphenylethyl)carbamate (Cpd PP, 9.94 g, 73% yield) as a solid. ¹ H NMR (400MHz, CDCl3) δ 8.18-8.29 (m, 2H), 7.24-7.32 (m, 2H), 7.11-7.17 (m, 3H), 5.28 (br.s., 1H), 3.45-3.53 (m, 2H), 2.85-2.92 (m, 2H), 2.32 (s, 3H).

A cooled (0°C) suspension of 4-nitrophenyl (5-chloro-2-methylphenethyl)carbamate (Cpd PP, 9.94 g, 29.7 mmol) in anhydrous 1,2-dichloroethane (120 mL) was treated with fresh trifluoromethanesulfonic acid (45.8 g, 305 mmol, 27.0 mL) and stirred at 0°C for 30 minutes. The reaction mixture was then heated to 70°C for 3 hours. After cooling to room temperature, the reaction mixture was carefully poured into ice water (200 mL) and stirred until the ice had completely melted. The biphasic mixture was extracted with dichloromethane (2 x 200 mL). The combined organic extracts were washed with 2M sodium carbonate (200 mL). The aqueous phase was back-extracted with dichloromethane (200 mL). The dichloromethane extracts were combined, dried over sodium sulfate, and concentrated to yield 8-chloro-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd QQ, 4.17 g, 72% yield) as a solid. ¹ H NMR (400 MHz, CDCl3) δ: 7.15-7.22 (m, 1H), 7.08 (d, J = 13.69 Hz, 1H), 6.46 (br. s., 1H), 3.43-3.51 (m, 2H), 2.88 (t, J = 6.17 Hz, 2H), 2.28 (s, 3H). MS: 196 [M+H] ⁺ .

A flask containing 8-chloro-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd QQ, 6.0 g, 30.7 mmol) was cooled in an ice bath. Concentrated sulfuric acid (125.0 mL) was added, and the mixture was stirred at 0°C for 30 minutes. Solid N-iodosuccinimide (20.7 g, 92.0 mmol) was added all at once, and the mixture was stirred at 0°C for 3 hours. The solution was carefully poured into ice water (300 mL), causing a precipitate to form. The suspension was extracted with ethyl acetate (300 mL) _. Both the organic and aqueous phases contained precipitates. The organic phase was washed with 10% _{Na₂S₂O₃} (300 _mL ) to remove excess iodine, and the aqueous layer was extracted with ethyl acetate (200 mL). The combined organic phases were dried over sodium sulfate and concentrated to dryness. The solid residue was stirred in methanol (100 mL). The insoluble material was collected by filtration, and the white solid precipitate (~14 g) was slurried in carbon disulfide (100 mL). The solid was collected by filtration, rinsed with carbon disulfide, and dried under vacuum to yield 8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd RR, 8.57 g, 87% yield) as a solid. ¹ H NMR (400 MHz, DMSO-d6) δ 8.15 (br. s., 1H), 7.94 (s, 1H), 3.26 (td, J = 6.17, 4.03 Hz, 2H), 2.75 (t, J = 6.24 Hz, 2H), 2.22 (s, 3H). MS: 321 [M = H] ⁺ .

Compound SS: 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dichloro- 3,4-Dihydroisoquinolin-1(2H)-one

A room temperature suspension of 7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd R, 14.9 g, 50.4 mmol) in ethyl acetate (300 mL) was treated with a 1.0 M solution of potassium tert-butoxide in THF (65.5 mL, 65.5 mmol), causing the solid to dissolve. After a few minutes, a precipitate began to form. 2-(Benzyloxy)-3-(Chloromethyl)-4-methoxy-6-methylpyridine (CpdKK, 14.0 g, 50.4 mmol) was added, and the resulting suspension was heated at 75°C for 4 hours. After cooling to room temperature, the mixture was washed with water (2x) and saturated aqueous NaCl solution, concentrated, and the residue was crystallized from ethyl acetate to give 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (CpdSS, 21.96 g, 81% yield) as a white solid. ¹ H NMR (400MHz, DMSO-d6) δ8.05 (s, 1H), 7.34-7.40 (m, 2H), 7.18-7.25 (m, 3H), 6.70 (s, 1H), 5.36 ( s, 2H), 4.68 (s, 2H), 3.83 (s, 3H), 3.16 (t, J=6.17Hz, 2H), 2.71 (t, J=6.17Hz, 2H), 2.38 (s, 3H). MS: 535, 537[M+H] ⁺ .

Compound TT: 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-8-chloro-7-iodo-5-methyl-3, 4-Dihydroisoquinolin-1(2H)-one

Solid potassium tert-butoxide (796 mg, 7.09 mmol) was added in several portions to a cooled (0°C) solution of 8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd RR, 1.52 g, 4.73 mmol) in anhydrous DMF (20 mL). Stirring was continued at 0°C for 30 minutes, followed by the dropwise addition of a solution of 2-(benzyloxy)-3-(chloromethyl)-4,6-dimethylpyridine (Cpd D, 1.18 g, 4.49 mmol) in anhydrous DMF (5 mL). After stirring at 0°C for 20 minutes, the mixture was poured into ice water (50 mL) and extracted with ethyl acetate (4 x 50 mL). The combined organic layers were washed with brine (4 x 50 mL), dried over sodium sulfate, filtered, and concentrated. The crude yellow solid (-2.4 g) was purified by silica gel chromatography (eluting with 5/1 petroleum ether/ethyl acetate) to afford 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd TT, 1.7 g, 66% yield) as an off-white solid. ¹ H NMR (400MHz, CDCl3) δ7.76 (s, 1H), 7.45 (d, J=7.2Hz, 2H), 7.36-7.30 (m, 3H), 6.62 (s, 1H), 5.42 (s, 2 H), 4.88 (s, 2H), 3.24 (t, J=6.2Hz, 2H), 2.50 (t, J=6Hz, 2H), 2.42 (s, 3H), 2.32 (s, 3H), 2.13 (s, 3H). MS: 547[M+H] ⁺ .

Compound UU: 2-((2-(Benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-iodo-5-methyl 3,4-dihydroisoquinolin-1(2H)-one

Solid potassium tert-butoxide (1.68 g, 14.9 mmol) was added in several portions to a cooled (0°C) solution of 8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd RR, 3.2 g, 9.95 mmol) in anhydrous DMF (50 mL). Stirring was continued at 0°C for 30 minutes, followed by the dropwise addition of a solution of 2-(benzyloxy)-3-(chloromethyl)-4-methoxy-6-methylpyridine (Cpd KK, 2.63 g, 14.9 mmol) in anhydrous DMF (50 mL). After stirring at 0°C for 30 minutes, the mixture was poured into ice water (100 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with brine (4 x 100 mL), dried over sodium sulfate, filtered, and concentrated. The crude yellow solid (~5 g) was purified by silica gel chromatography (eluting with 20 to 50% ethyl acetate in petroleum ether). The resulting product was dissolved in dichloromethane (10 mL), added to petroleum ether (30 mL), and stirred at room temperature until a precipitate formed (30 minutes). The precipitate was collected by filtration and dried to a white solid. TLC of the precipitate showed that impurities still existed, so it was purified again by silica gel chromatography (eluting with 0 to 10% methanol in dichloromethane) to provide 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd UU, 2.8 g, 50% yield) as a white solid. ¹ H NMR (400MHz, CDCl3) δ7.75 (s, 1H), 7.42-7.40 (m, 2H), 7.29-7.27 (m, 1H), 7.25-7.22 (m, 2H), 6.38 (s, 1H), 5. 42 (s, 2H), 4.87 (s, 2H), 3.83 (s, 3H), 3.14 (t, J=6.2Hz, 2H), 2.50 (t, J=6.2Hz, 2H), 2.44 (s, 3H), 2.13 (s, 3H). MS: 563[M+H] ⁺ .

Example

General Methods and Representative Examples

Method A

Example 1: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R)-2-hydroxy-1-[(3R)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one

Example 2: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R ^* )-2-hydroxy-1-[(3S ^* )-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one

Example 3: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1S ^* )-2-hydroxy-1-[(3R ^* )-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one

Example 4: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1S)-2-hydroxy-1-[(3S)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one

A cooled (0°C) solution of methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (Cpd T, 800 mg, 0.487 mmol) in anhydrous N,N-dimethylformamide (70 mL) was treated with sodium hydride (60 wt% dispersion in mineral oil, 125 mg, 3.12 mmol) and stirred at 10°C for 15 minutes. The mixture was cooled to 0°C again and 3-iodotetrahydrofuran (463 mg, 2.34 mmol) was added. After stirring at room temperature for 12 hours, glacial acetic acid (2 drops) and water (10 mL) were added, and the mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic extracts were washed with brine (10 mL), dried over sodium sulfate, concentrated, and purified by column chromatography (silica gel, petroleum ether/EtOAc = 5:1 to 1:1) to give 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-3-yl)acetic acid (1a, 500 mg, 56.3%) as a white solid; and methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-3-yl)acetate (1b, 150 mg, 16.5%) as a yellow solid.

A solution of 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-3-yl)acetic acid (1a, 650 mg, 1.14 mmol), potassium carbonate (315 mg, 2.28 mmol), and iodomethane (324 mg, 2.28 mmol) in N,N-dimethylformamide (8 mL) was stirred at room temperature for 12 hours. The mixture was diluted with water (20 mL) and ethyl acetate (50 mL). The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The organic layers were combined, washed with brine (3×10 mL), dried over sodium sulfate, filtered, and concentrated under vacuum to give a crude product, which was purified by column chromatography (silica gel, petroleum ether/EtOAc=1:1) to give methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-3-yl)acetate (1b, 600 mg, 90.1%) as a white solid.

Lithium borohydride (28 mg, 1.29 mmol) was added all at once to a room temperature solution of methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-3-yl)acetate (1b, 250 mg, 0.428 mmol) in anhydrous tetrahydrofuran (25 mL). The resulting mixture was heated at 60°C for 2 hours. The mixture was quenched with water (5 mL) and extracted with ethyl acetate (3 x 15 mL). The organic layers were combined, washed with brine (5 mL), dried over sodium sulfate, filtered, and concentrated under vacuum to give a crude product, which was purified by preparative TLC (petroleum ether/EtOAc=1:1) to give 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(2-hydroxy-1-(tetrahydrofuran-3-yl)ethyl)-3,4-dihydroisoquinolin-1(2H)-one as a white solid (a mixture of 4 stereoisomers, 160 mg, 67.2%). Combined batches (500 mg total) of this stereoisomer mixture were separated by preparative chiral SFC (Chiralpak AD, 250 x 30 mm ID, 5 μm, mobile phase 35% EtOH NH ₃ H ₂ O, flow rate 50 mL/min) to afford the isolated isomers 1c (peak 1, 80 mg, 15.9%), 2c (peak 2, 90 mg, 17.9%), 3c (peak 3, 110 mg, 21.9%), and 4c (peak 4, 100 mg, 19.9%) as white solids. The absolute stereochemistry of the individual isomers was not determined at this stage.

A solution of 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(2-hydroxy-1-(tetrahydrofuran-3-yl)ethyl)-3,4-dihydroisoquinolin-1(2H)-one stereoisomer 1c (80 mg, 0.144 mmol) in dichloromethane (3 mL) and trifluoroacetic acid (3 mL) was stirred at 35°C for 5 hours and then evaporated to dryness. The residue was taken up in methanol (10 mL), cooled to 10°C, and potassium carbonate (99.5 mg, 0.720 mmol) was added. The mixture was stirred at 10°C for 30 minutes, filtered, and the filter pad was rinsed with dichloromethane/methanol (10:1, 10 mL). The filtrate was concentrated in vacuo, and the residue was purified by preparative TLC (CH ₂ Cl ₂ /MeOH=10:1, Rf=0.4 in CH ₂ Cl ₂ /MeOH=10:1) to give Example 1 (38 mg, 57%) as a white solid.

By the same procedure, stereoisomer 2c (90 mg, 0.162 mmol) gave Example 2 as a white solid (44 mg, 59%); stereoisomer 3c (110 mg, 0.198 mmol) gave Example 3 as a white solid (61 mg, 66%); and stereoisomer 4c (100 mg, 0.18 mmol) gave Example 4 as a white solid (35 mg, 41%).

The small molecule X-ray crystal structure of Example 4 showed absolute (S,S) stereochemistry. Since the ^1H NMR spectrum was identical to that of Example 4, Example 1 had absolute (R,R) stereochemistry. The ^1H NMR spectra of Examples 2 and 3 were identical to each other and significantly different from that of Example 4, indicating that they were (R,S) and (S,R) stereoisomers, although the absolute configuration of each was not determined.

Example 1: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R)-2-hydroxy-1-[(3R)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one (absolute stereochemistry assigned based on the crystal structure of the enantiomeric mixture). ¹ H NMR (400MHz, CD3OD) δ7.63 (s, 1H), 6.12 (s, 1H), 4.77 (s, 2H), 3.94-3.90 (m, 1H), 3.81-3.80 ( m, 3H), 3.59-3.57 (m, 2H), 3.51-3.49 (m, 2H), 3.17-3.10 (m, 1H), 2.98-2.95 (m, 2H), 2.71 (br s, 1H), 2.30 (s, 3H), 2.29-2.25 (m, 1H), 2.25 (s, 3H), 1.83-1.78 (m, 1H). MS: 465[M+H] ⁺ . Chiral analysis: 95.66% ee/de; retention time: 6.867 minutes; column: Chiralpak AD-3 150×4.6 mm ID, 3 μm; mobile phase: 5% to 40% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.5 ml/min; wavelength 220 nm.

Example 2: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R ^* )-2-hydroxy-1-[(3S ^* )-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one (relative stereochemistry known, absolute stereochemistry not determined). ¹ H NMR (400MHz, CD3OD) δ7.62 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 4.13-4.11 (m, 1H), 3.78-3.75 ( m, 1H), 3.69-3.68 (m, 2H), 3.61-3.59 (m, 3H), 3.51-3.50 (m, 2H), 2.98-2.95 (m, 2H), 2.65 (br s, 1H), 2.30 (s, 3H), 2.25 (s, 3H), 1.77-1.75 (m, 1H), 1.42-1.37 (m, 1H). ). MS: 465[M+H] ⁺ . Chiral analysis: 98.70% ee/de; retention time: 7.309 minutes; column: Chiralpak AD-3 150×4.6 mm ID, 3 μm; mobile phase: 5% to 40% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.5 ml/min; wavelength 220 nm.

Example 3: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1S ^* )-2-hydroxy-1-[(3R ^* )-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one (relative stereochemistry known, absolute stereochemistry not determined). ¹ H NMR (400MHz, CD3OD) δ7.62 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 4.12-4.11 (m, 1H), 3.80-3.78 ( m, 1H), 3.69-3.67 (m, 3H), 3.67-3.62 (m, 2H), 3.61-3.50 (m, 2H), 2.98-2.95 (m, 2H), 2.65 (br s, 1H), 2.29 (s, 3H), 2.25 (s, 3H), 1.77-1.74 (m, 1H), 1.42-1.37 (m, 1H). MS: 465[M+H] ⁺ . Chiral analysis: 96.48% ee/de; retention time: 8.021 minutes; column: Chiralpak AD-3 150×4.6 mm ID, 3 μm; mobile phase: 5% to 40% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.5 ml/min; wavelength 220 nm.

Example 4: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1S)-2-hydroxy-1-[(3S)-tetrahydrofuran-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one (absolute stereochemistry determined by X-ray crystal structure). ¹ H NMR (400MHz, CD3OD) δ7.64 (s, 1H), 6.13 (s, 1H), 4.78 (s, 2H), 3.95-3.90 (m, 1H), 3.83-3.81 ( m, 3H), 3.60-3.55 (m, 2H), 3.55-3.52 (m, 2H), 3.32-3.19, (m, 1H), 2.99-2.96 (m, 2H), 2.75 (br s, 1H), 2.32 (s, 3H), 2.31-2.29 (m, 1H), 2.27 (s, 3H), 1.84-1.79 (m, 1H). MS: 465[M+H] ⁺ . Chiral analysis: 99.18% ee/de; retention time: 8.429 minutes; column: Chiralpak AD-3 150×4.6 mm ID, 3 μm; mobile phase: 5% to 40% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.5 ml/min; wavelength 220 nm.

Method B

Example 5: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H)-one

Example 6: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1ξ)-1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H)-one - Isomer A

Example 7: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1ξ)-1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H)-one - Isomer B

HBr (800 μL, 4.42 mmol, 33% wt in HOAc) was added to an ice-bath cooled solution of methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate (Cpd U, 800 mg, 1.48 mmol) in anhydrous dichloromethane (10 mL), resulting in gas evolution. The solution was allowed to warm to room temperature and stirred overnight. The reaction mixture was carefully quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate (2×50 mL). The combined organic phases were dried over sodium sulfate, concentrated to dryness, and purified by silica gel column chromatography (eluting with a gradient of 0 to 10% MeOH/EA) to give racemic methyl 2-bromo-2-(5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (5a, 655 mg, 88%) as a solid. MS:501.00/502.05. ^1H NMR (400MHz, CDCl3) δ7.91 (s, 1H), 6.18 (s, 1H), 6.03 (s, 1H), 4.76 (s, 2H), 3.81 ( s, 3H), 3.68 (t, J=6.24Hz, 2H), 2.98 (t, J=6.24Hz, 2H), 2.45 (s, 3H), 2.37 (s, 3H).

A mixture of methyl 2-bromo-2-(5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)acetate (5a, 162 mg, 0.323 mmol), (3R)-3-fluoropyrrolidine hydrochloride (144 mg, 1.15 mmol), N,N-diisopropylethylamine (0.35 mL, 2.01 mmol), and anhydrous N,N-dimethylformamide (4 mL) was stirred at room temperature overnight. The reaction mixture was extracted by partitioning with ethyl acetate (20 mL) and water (20 mL). The organic phase was separated, washed sequentially with water (20 mL) and brine (20 mL), dried over sodium sulfate, and concentrated to dryness to afford crude methyl 2-(5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-((R)-3-fluoropyrrolidin-1-yl)acetate as a mixture of diastereomers (5b, 162 mg, 98% yield). This material was used in the next step without further purification. LCMS: T = 510.15/511.10/512.20.

A mixture of crude diastereoisomers of methyl 2-(5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-((R)-3-fluoropyrrolidin-1-yl)acetate (5b, 152 mg, 0.298 mmol) and anhydrous tetrahydrofuran (4.0 mL) were treated with lithium borohydride (2.0 M in THF, 0.45 mL, 0.90 mmol) followed by a few drops of methanol. This addition was repeated four times before quenching the reaction with 2 M _NH4Cl (20 mL) and extracting with ethyl acetate (2 x 20 mL). The combined organic phases were dried over sodium sulfate, concentrated to dryness, and purified by preparative HPLC to give 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H)-one as a mixture of diastereomers at the benzyl carbon (Example 5, 32.2 mg, ²² % yield over two steps). NMR (400MHz, DMSO-d6) δ11.66 (br.s., 1H), 7.80 (d, J=3.67Hz, 1H), 6.00 (s, 1H), 5 .15-5.43(m, 1H), 4.85(br.s., 1H), 4.69(s, 2H), 4.03-4.13(m, 1H), 3.66-3.84(m, 2H), 3.50-3.63(m, 2H), 3.02-3.10(m, 1H), 2.94-3.02(m, 2H), 2.71-2.90(m, 2H), 2 .42-2.55(m, 1H), 2.28(s, 3H), 2.24(s, 3H), 2.09-2.23(m, 1H), 1.87-2.08(m, 1H). MS: 482[M+H] ⁺ .

A mixture of diastereomers of 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H)-one (Example 5, 20.0 mg, 0.0415 mmol) was separated by chiral preparative SFC (Chiralcel OJ-3 4.6 x 100 mm 3μ column, eluted with 10% MeOH/DEA @ 120 bar, 4 mL/min). After freeze-drying, Example 6 (peak 1, retention time 1.18 minutes, 5.65 mg, 28%) and Example 7 (peak 2, retention time 1.42 minutes, 6.23 mg, 31%) were obtained. The absolute stereochemistry of the benzylic carbon in each isomer was not determined.

Example 6: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1ξ)-1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H)-one—Isomer A. MS: 482 [M+H] ⁺ . Chiral analysis: ∼88% de, [α]D = +62.1° (c 0.01 MeOH)

Example 7: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1ξ)-1-[(3R)-3-fluoropyrrolidin-1-yl]-2-hydroxyethyl}-3,4-dihydroisoquinolin-1(2H)-one—Isomer B. MS: 482 [M+H] ⁺ . Chiral analysis: ~98% de; [α]D = –58.9° (c 0.01 MeOH)

Method C

Example 8: (+)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{fluoro[1-(hydroxyacetyl)piperidin-4-yl]methyl}-3,4-dihydroisoquinolin-1(2H)-one

Example 9: (-)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{fluoro[1-(hydroxyacetyl)piperidin-4-yl]methyl}-3,4-dihydroisoquinolin-1(2H)-one

iPrMgCl-LiCl (1.3 M in THF, 0.850 mL, 1.10 mmol) was added to a solution of 2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd S, 311.0 mg, 0.598 mmol) in tetrahydrofuran (5.0 mL) and 1,4-dioxane (0.5 mL) at -40°C (in an acetonitrile/dry ice bath). The reaction mixture was stirred for 1 hour. N-Boc-4-formylpiperidine (0.242 g, 1.14 mmol) was then added, and the flask was allowed to warm to 0°C in an ice bath. After maintaining the temperature at 0°C for 1 hour, the reaction was quenched with saturated aqueous _NH4Cl and extracted with MTBE. The MTBE layer was concentrated and the resulting oil was purified on silica gel (Isco RediSepRf, 12 g, eluting with a gradient of 10 to 70% ethyl acetate in heptane) to provide racemic tert-butyl 4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)(hydroxy)methyl)piperidine-1-carboxylate (8a, 0.229 g, 59%) ^{as a} white solid. HNMR (400MHz, CD3OD) δ7.66 (s, 1H), 7.36-7.44 (m, 2H), 7.18-7.31 (m, 3H), 6. 71 (s, 1H), 5.42 (s, 2H), 5.04 (d, J = 5.14Hz, 1H), 4.83 (d, J = 1.96Hz, 2H), 4.08 ( d, J=12.72Hz, 2H), 3.23 (t, J=6.24Hz, 2H), 2.73 (t, J=6.11Hz, 2H), 2.53-2.7 1(m, 2H), 2.39(s, 3H), 2.35(s, 3H), 1.76-1.90(m, 1H), 1.32-1.62(m, 13H); MS 654,656[M+H] ⁺ .

Deoxo-Fluor ^™ (50% in THF, 0.165 mL, 0.39 mmol) was added to a solution of racemic tert-butyl 4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)(hydroxy)methyl)piperidine-1-carboxylate (8a, 88 mg, 0.13 mmol) in dichloromethane (4 mL) cooled in a dry ice/acetone bath. The mixture was stirred at -78°C for 5 minutes, the cooling bath was removed, and the mixture was stirred for 5 minutes. The reaction was quenched by the addition of saturated aqueous _NaHCO₃ , the layers were separated, and the dichloromethane layer was concentrated. The resulting oil was purified on silica gel (Biotage SNAP, HP-Sil, 10 g, eluting with a gradient of 0 to 40% ethyl acetate in heptane) to provide racemic tert-butyl 4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)fluoromethyl)piperidine-1-carboxylate (8b, 0.075 g, 85%) as a white, sticky solid. MS: 656, 658 [M+H] ⁺ .

A solution of racemic tert-butyl 4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)fluoromethyl)piperidine-1-carboxylate (8b, 0.075 g, 0.11 mmol) in trifluoroacetic acid (5.0 mL) was heated to 50°C for 1 hour. The reaction mixture was diluted with heptane and concentrated under vacuum. The residue was taken up in ethanol and concentrated. The remaining white solid was dried under vacuum to provide crude racemic 5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-7-(fluoro(piperidin-4-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (8c, 0.085 g, TFA salt). MS 466, 468 [M+H] ⁺ . This material was dissolved in dichloromethane (5 mL) and cooled to 0°C. Triethylamine (0.060 mL, 0.43 mmol) and 2-acetoxyacetyl chloride (0.017 mL, 0.16 mmol) were added sequentially, and the mixture was stirred for 30 minutes, followed by the addition of a few drops of methanol to quench excess reagent. The solution was concentrated under vacuum, and the residue was dissolved in methanol (5 mL) and treated with potassium carbonate (0.100 g, 0.724 mmol). After 4 hours at room temperature, the reaction mixture was filtered, concentrated, and purified by preparative chiral SFC (OJ-H, 21 x 250 mm column, 32 mL MeOH:8 mL CO ₂ , 100 bar, 40 mL/min) to provide Example 8 (Peak 1, 9.6 mg, 15%) and Example 9 (Peak 2, 8.1 mg, 13%) as white solids, separated enantiomers. The absolute stereochemistry of the individual isomers has not been determined, but optical rotation measurements have been obtained.

Example 8: (+)-5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{fluoro[1-(hydroxyacetyl)piperidin-4-yl]methyl}-3,4-dihydroisoquinolin-1(2H)-one. MS 524, 526 [M+H] ⁺ . Optical rotation: [α]D = +9.9° (c, 0.1, DMSO). Chiral analysis: >99% ee; retention time 8.13 minutes, Lux Cellulose-2 4.6 × 100 mm 3U column, 60% MeOH @ 120 bar, 4 mL/min.

Example 9: (-)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{fluoro[1-(hydroxyacetyl)piperidin-4-yl]methyl}-3,4-dihydroisoquinolin-1(2H)-one. ¹ H NMR (600 MHz, DMSO-d ₆ ) δ11.55 (s, 1H), 7.58 (d, J = 4.95Hz, 1H), 5.88 (s, 1H), 5.72-5.84 (m, 1H), 4.55 (q, J = 13.75H z, 2H), 4.46 (br.s., 1H), 4.30-4.42 (m, 1H), 4.05-4.13 (m, 1H), 3.97-4.04 (m, 1H), 3.69 (t, d, J = 18.71 Hz, 1H), 2.11 (s, 3H), 1.61 (br. s., 1H), 1.46 (br. s., 1H), 1.34-1.42 (m, 1H), 1.23 (s, 5H); MS 524, 526 [M+H] ⁺ . Optical rotation: [α] = –6.5° (c, 0.1, DMSO). Chiral analysis: ~95% ee; retention time 10.29 min, Lux Cellulose-2 4.6 × 100 mm 3u column, 60% MeOH @ 120 bar, 4 mL/min.

Method D

Example 10: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one—Isomer A

Example 11: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one—Isomer B

A solution of methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate (Cpd U, 150 mg, 0.278 mmol) in dichloromethane (5 mL) was added dropwise to a stirred solution of anhydrous methanol (12 mg, 0.36 mmol) and dirhodium tetraacetate (1.2 mg, 0.0028 mmol) in dichloromethane (5 mL) at room temperature over 60 minutes. After the addition was complete, the reaction mixture was heated at reflux for 18 hours. The mixture was concentrated and purified by flash chromatography (eluted with petroleum ether/EtOAc = 10:1, Rf ~ 0.3) to give racemic 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-methoxyacetic acid methyl ester (11a, 100 mg, 66%) as a brown oil.

Solid lithium borohydride (12 mg, 0.55 mmol) was added all at once to a stirred solution of racemic methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-methoxyacetate (11a, 100 mg, 0.184 mmol) in anhydrous tetrahydrofuran (10 mL) at room temperature. The resulting mixture was heated at 60°C for 1 hour. The reaction was quenched with water (15 mL) and extracted with ethyl acetate (3 x 15 mL). The combined organic extracts were washed with saturated aqueous NaCl (15 mL), dried over sodium sulfate, filtered, and concentrated to afford the crude product (91 mg). Purification by flash chromatography (elution with petroleum ether/EtOAc = 1:1) gave racemic 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (11b, 60 mg, 63%) as a white solid.

A solution of racemic 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (11b, 60 mg, 0.12 mmol) in dichloromethane (2 mL) and trifluoroacetic acid (2 mL) was stirred at room temperature for 18 hours. LC-MS analysis indicated complete deprotection of the benzyl group, but some trifluoroacetate was present at the pyridinone oxygen. Therefore, the mixture was concentrated, and methanol (10 mL) and potassium carbonate (80.9 mg, 0.585 mmol) were added. The resulting mixture was stirred at room temperature for 30 minutes, at which point LC-MS analysis indicated complete deprotection of the TFA ester. The mixture was filtered, and the collected solid was washed with 10:1 DCM/MeOH (10 mL). The filtrate was concentrated. Purification by flash chromatography (elution with DCM/MeOH = 10:1, Rf ~ 0.6) gave racemic 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (37 mg, 75%) as a white solid.

Combined batches of racemic 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (100 mg, 0.235 mmol total) were separated by chiral SFC (Chiralpak IC 250 mm × 30 mm, 10 μm column, eluted with 50% EtOH/ _NH₄OH , flow rate 70 mL/min). After freeze-drying, Example 10 (Peak 1, 33 mg, 33%) and Example 11 (Peak 2, 35 mg, 35%) were obtained as off-white solids. The absolute stereochemistry of the individual isomers was not determined.

Example 10: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one—Isomer A. ¹ H NMR (400MHz, CDCl3) δ12.23 (br.s., 1H), 7.53 (s, 1H), 5.95 (s, 1H), 4.92-4.89 (m, 1H), 4.78 (s, 2H), 3.80-3.7 1 (m, 1H), 3.68-3.62 (m, 2H), 3.53-3.51 (m, 1H), 3.33 (s, 3H), 2.94 (t, J=6.0Hz, 2H), 2.35 (s, 3H), 2.29 (s, 3H). MS: 425[M+H] ⁺ . Chiral analysis: 100% ee, retention time 7.717 minutes, column: Chiralpak IC-3 150×4.6 mm ID, 3 μm; mobile phase: 40% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.35 mL/min.

Example 11: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(2-hydroxy-1-methoxyethyl)-3,4-dihydroisoquinolin-1(2H)-one—Isomer B. ¹ H NMR (400MHz, CDCl3) δ12.15 (br.s., 1H), 7.53 (s, 1H), 5.95 (s, 1H), 4.92-4.89 (m, 1H), 4.78 (s, 2H), 3.80-3.7 1 (m, 1H), 3.68-3.63 (m, 2H), 3.55-3.51 (m, 1H), 3.34 (s, 3H), 2.94 (t, J=5.6Hz, 2H), 2.35 (s, 3H), 2.29 (s, 3H). MS: 425[M+H] ⁺ . Chiral analysis: 100% ee, retention time 11.063 minutes, column: Chiralpak IC-3 150×4.6 mm ID, 3 μm; mobile phase: 40% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.35 mL/min.

Method E

Example 12: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R)-2-hydroxy-1-[(2S)-tetrahydrofuran-2-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one

Example 13: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R)-2-hydroxy-1-[(2R)-tetrahydrofuran-2-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one

A solution of copper(II) trifluoromethanesulfonate (120 mg, 0.332 mmol) and (S)-(-)-2,2'-isopropylidene-bis(4-phenyl-2-oxazoline) (130 mg, 0.389 mmol) in anhydrous tetrahydrofuran (4 mL) was added to a solution of methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-diazoacetate (Cpd U, 1.78 g, 3.29 mmol) in anhydrous tetrahydrofuran (20 mL). The resulting solution was heated under reflux in an 80°C oil bath overnight. After cooling to room temperature, the reaction mixture was concentrated to dryness and purified on a silica gel column (eluting with a gradient of 0 to 40% EA/HEP) to provide a mixture of diastereomers with (S) geometry at the benzylic carbon (stereochemistry assigned similarly to Jiménez-Osés, G. et al., J. Org. Chem. 2013, 78, 5851-5857), (2S)-methyl 2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-2-yl)acetate (12a, 333 mg, 17%). MS: 583.10/584.20 [M+H] ⁺ . ¹ H NMR (400MHz, CDCl3) δ7.69 (d, J=2.20Hz, 1H), 7.45 (d, J=7.58Hz, 2H), 7.30-7.39 (m, 3H ), 6.64(s, 1H), 5.45(s, 2H), 4.80-4.92(m, 2H), 4.41-4.59(m, 1H), 3.79-4.02(m, 1H), 3 .72-3.80(m, 1H), 3.70(d, J=5.01Hz, 3H), 3.23-3.31(m, 2H), 2.69-2.76(m, 2H), 2.44(s , 3H), 2.34 (d, J=3.79Hz, 3H), 1.93-2.18 (m, 1H), 1.80-1.94 (m, 2H), 1.55-1.80 (m, 2H).

[Under the same conditions, the use of the enantiomeric (R)-(-)-2,2'-isopropylidene-bis(4-phenyl-2-oxazoline) ligand produced a mixture of diastereomers with (R) geometry at the benzyl position, methyl (2R)-2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-2-yl)acetate.]

Lithium borohydride (2.0 M in THF, 1.0 mL, 2.0 mmol) was added to a solution of methyl (2S)-2-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-2-(tetrahydrofuran-2-yl)acetate (12a, 333 mg, 0.571 mmol) in anhydrous tetrahydrofuran (10 mL), followed by a few drops of methanol. Gas evolution occurred. The mixture was stirred at room temperature for 1 hour, then quenched with 2 M _NH₄Cl (10 mL), diluted with water, and extracted with diethyl ether (3×50 mL). The combined organic phases were dried over sodium sulfate and concentrated to dryness to provide crude 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-((1R)-2-hydroxy-1-(tetrahydrofuran-2-yl)ethyl)-3,4-dihydroisoquinolin-1(2H)-one (12b, 280 mg, 88%) as a mixture of diastereomers with (R) geometry at the benzylic carbon. This mixture was used in the next step without further purification. MS: 555.20/557.20.

A solution of the crude diastereoisomer mixture, 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-((1R)-2-hydroxy-1-(tetrahydrofuran-2-yl)ethyl)-3,4-dihydroisoquinolin-1(2H)-one (12b, 280 mg, 0.504 mmol) in trifluoroacetic acid (8 mL) was stirred at 50°C for 1 hour. After removal of excess trifluoroacetic acid, the residue was dissolved in methanol (10 mL) and treated with 4M NaOH at 50°C for 30 minutes. The reaction mixture was partitioned and extracted with ethyl acetate (50 mL) and water (50 mL). The organic phase was separated. The aqueous phase was acidified to pH ~2 to 3 and extracted with ethyl acetate (2 x 50 mL). The combined organic phases were dried over sodium sulfate, concentrated to dryness, and purified by chiral SFC (Chiralpak AD-3 4.6 x 100 mm 3μ column; eluted with 5 to 60% MeOH over 3 minutes at 120 bar and 4 mL/min) to afford the separated diastereomeric products, Example 12 (peak 1, 32 mg, 14%) and Example 13 (peak 2, 77 mg, 33%). The stereochemistry of the isolated products was determined similarly to that described by Jiménez-Osés, G. et al., J. Org. Chem. 2013, 78, 5851-5857.

Example 12: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R)-2-hydroxy-1-[(2S)-tetrahydrofuran-2-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one. ¹ H NMR (400MHz, CD3OD) δ7.58 (s, 1H), 6.11 (s, 1H), 4.76 (s, 2H), 4.10-4.21 (m, 1H), 3.86-3.99 (m, 3H), 3.75-3.84 (m, 1H), 3 .64-3.72(m, 1H), 3.46-3.55(m, 2H), 2.91-3.01(m, 2H), 2.29(s, 3H), 2.25(s, 3H), 1.73-1.97(m, 3H), 1.44-1.58(m, 1H). MS: 465{M+H] ⁺ . Chiral analysis: 91% ee/de; retention time 2.91 minutes, Chiralpak AD-3 4.6×100 mm 3μ column; eluted with 5 to 60% MeOH over 3 minutes, 120 bar, 4 ml/min.

Example 13: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(1R)-2-hydroxy-1-[(2R)-tetrahydrofuran-2-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one. ¹ H NMR (400MHz, CD3OD) δ7.66 (s, 1H), 6.10 (s, 1H), 4.77 (s, 2H), 4.27 (dt, J=7.86, 6.16Hz, 1H), 3.72-3.91 (m, 3H), 3.67 (t, J=6.79Hz, 2H ), 3.49 (t, J=6.24Hz, 2H), 2.95 (t, J=6.17Hz, 2H), 2.28 (s, 3H), 2.24 (s, 3H), 2.01-2.13 (m, 1H), 1.68-1.92 (m, 2H), 1.49-1.62 (m, 1H). MS: 465{M+H] ⁺ . Chiral analysis: 93% ee/de; retention time 3.21 minutes, Chiralpak AD-3 4.6×100 mm 3μ column; eluted with 5 to 60% MeOH over 3 minutes, 120 bar, 4 ml/min.

Method F

Example 14: 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(propan-2-yl)-3,4-dihydroisoquinolin-1(2H)-one

A solution of 2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd S, 500 mg, 0.961 mmol), triethylamine (0.30 mL, 2.2 mmol) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II)-dichloromethane complex (24 mg, 0.028 mmol) in 2-propanol (15 mL) in a capped microwave tube was heated at 100° C. for 1 hour in a microwave reactor under nitrogen. After removal of the solvent, the product was extracted with diethyl ether (3 x 10 mL) and the combined organic extracts were washed with water (2x), dried over magnesium sulfate, concentrated, and purified by flash chromatography (silica gel, 0 to 60% EtOAc in heptane) to give 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(prop-1-en-2-yl)-3,4-dihydroisoquinolin-1(2H)-one (14a, 240 mg, 52%) as a white foam. ^1H NMR (400MHz, CDCl3) δ7.37 (d, J=6.8Hz, 2H), 7.30-7.20 (m, 3H), 7.18 (d, J=2.6Hz, 1H), 6.55 (s, 1H), 5.35 (s, 2H), 5.16 (d, J=1.5H z, 1H), 4.85 (d, J=1.5Hz, 1H), 4.80 (s, 2H), 3.20 (t, J=6.2Hz, 2H), 2.65 (t, J=6.2Hz, 2H), 2.34 (s, 3H), 2.26 (s, 3H), 2.01 (s, 3H). MS: 481[M+H] ⁺ .

A solution of 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(prop-1-en-2-yl)-3,4-dihydroisoquinolin-1(2H)-one (14a, 75 mg, 0.16 mmol) and platinum (IV) oxide (71 mg, 0.31 mmol) in ethyl acetate (5 mL) was stirred under hydrogen balloon pressure for 2 hours. The catalyst was filtered off, and the solvent was removed in vacuo. The crude product was purified by supercritical fluid chromatography (SFC/ZymorSpher HAP 150×21.2 mm column, 8% MeOH @ 100 bar, 58 mL/min) to give 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(propan-2-yl)-3,4-dihydroisoquinolin-1(2H)-one (Example 14, 7 mg, 10%) as a white solid. ¹ H NMR (400MHz, CDCl3) δ7.35 (s, 1H), 5.90 (s, 1H), 4.78 (s, 2H), 3.63 (t, J=6.30Hz, 2H), 3. 5-3.6 (m, 1H), 2.90 (t, J=6.11Hz, 2H), 2.35 (s, 3H), 2.24 (s, 3H), 1.24 (d, J=6.85Hz, 6H). MS: 393[M+H].

Method G

Example 15: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(1-methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one—Isomer A

Example 16: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(1-methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one—Isomer B

A mixture of 2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd S, 200 mg, 0.384 mmol), (Z)-3-hexenyl-3-boronic acid catechol ester (155 mg, 0.769 mmol), cesium fluoride (182 mg, 1.20 mmol), dioxane (2 mL), and water (0.4 mL) was degassed with nitrogen for 5 minutes. Tetrakis(triphenylphosphine)palladium(0) (44.4 mg, 0.0384 mmol) was added, and the mixture was degassed with nitrogen and then heated to 100°C for 4 hours. The mixture was diluted with water (15 mL) and extracted with EtOAc (3×15 mL). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate, concentrated, and purified by column chromatography (silica gel, petroleum ether/EtOAc = 10:1, Rf ~ 0.5) to give (E)-2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(hex-3-en-3-yl)-3,4-dihydroisoquinolin-1(2H)-one (15a, 150 mg, 74.5%) as a colorless oil. ¹ H NMR (400MHz, CDCl3) δ7.45 (d, J=7.7Hz, 2H), 7.37-7.34 (m, 3H), 7.19 (s, 1H ), 6.62(s, 1H), 5.43(s, 2H), 5.28(t, J＝6.8Hz, 1H), 4.88(s, 2H), 3.28(t, J＝ 6.4Hz, 2H), 2.73 (t, J=6.4Hz, 2H), 2.45-2.43 (m, 2H), 2.42 (s, 3H), 2.33 (s , 3H), 2.19 (quint, J=7.6Hz, 2H), 1.04 (t, J=7.6Hz, 3H), 0.89-0.86 (m, 4H).

A stirred solution of (E)-2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(hex-3-en-3-yl)-3,4-dihydroisoquinolin-1(2H)-one (15a, 864 mg, 1.65 mmol) in methanol (46 mL) was bubbled with ozone at -78°C until a light purple color developed (~10 minutes). Nitrogen was bubbled through the solution until the purple color disappeared, followed by the addition of dimethyl sulfide (0.4 mL), and the solution was stirred at 10°C for 3 hours. The reaction mixture was concentrated under vacuum, and the residue was purified by column chromatography (silica gel, petroleum ether/EtOAc = 6:1, Rf ~ 0.5) to give 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-propionyl-3,4-dihydroisoquinolin-1(2H)-one (15b, 346 mg, 42.1%) as a yellow oil.

Sodium borohydride (52.6 mg, 1.39 mmol) was added to a cooled (0°C) solution of 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-propionyl-3,4-dihydroisoquinolin-1(2H)-one (15b, 346 mg, 0.696 mmol) in methanol (20 mL). Stirring was continued at 10°C for 1 hour. The reaction was quenched with saturated NH4Cl (30 mL) and extracted with ethyl acetate (3 x 25 mL). The combined organic layers were washed with brine (25 mL), dried over sodium sulfate, and concentrated under vacuum. The residue was purified by column chromatography (silica gel, petroleum ether/EtOAc = 2:1, Rf ~ 0.6) to give racemic 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-hydroxypropyl)-3,4-dihydroisoquinolin-1(2H)-one (15c, 300 mg, 86.4%) as a colorless oil.

Potassium tert-butoxide (111 mg, 0.985 mmol) and iodomethane (140 mg, 0.985 mmol) were added to a cooled (10° C.) solution of 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-hydroxypropyl)-3,4-dihydroisoquinolin-1(2H)-one (15c, 246 mg, 0.493 mmol) in N,N-dimethylformamide (25 mL) and stirred overnight at 10° C. The reaction was quenched with water (30 mL) and extracted with EtOAc (3×35 mL). The combined organic layers were washed with brine (35 mL), dried over sodium sulfate, concentrated, and purified by column chromatography (silica gel, petroleum ether/EtOAc = 5:1) to give racemic 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one (15d, 148 mg, 58.5%) as a yellow oil.

Trifluoroacetic acid (9 mL) was added dropwise to a stirred solution of 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one (15d, 218 mg, 0.423 mmol) in dichloromethane (9 mL) at 10° C. The resulting mixture was stirred at 25° C. overnight. The mixture was concentrated under vacuum, and the residue was purified by column chromatography (silica gel, _CH2Cl2 / _MeOH =10:1, Rf~0.55) to give racemic 5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-7-(1-methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one (15e, 150 mg, 83.8%) as a pink oil. The racemic mixture was separated by preparative SFC [column: (AD (250 mm*30 mm, 5 μm)), mobile phase: 25% MeOH NH ₃ H ₂ O, flow rate: 50 ml/min, wavelength: 220 nm, treatment: freeze drying] to give white solid isomer A (Example 15, 45.65 mg, 31.4%) and off-white solid isomer B (Example 16, 31.8 mg, 21.2%). The absolute stereochemistry of each isomer was not determined.

Example 15: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(1-methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one—Isomer A. ¹ H NMR (400MHz, CDCl3) δ11.88 (br s, 1H), 7.53 (s, 1H), 5.94 (s, 1H), 4.78 (s, 2H), 4.69-4.66 (m, 1H), 3.65 (t, J=4.8Hz, 2H), 3.24 (s, 3H), 2.9 3 (t, J=6.2Hz, 2H), 2.36 (s, 3H), 2.29 (s, 3H), 1.76-1.72 (m, 1H), 1.65-1.60 (m, 1H), 0.96 (t, J=7.0Hz, 3H). MS: 423[M+H] ⁺ . Chiral analysis: 100% ee; column: Chiralpak AD-H 250×4.6 mm ID, 5 μm; retention time: 8.04 minutes; mobile phase: 5% to 40% methanol (0.05% DEA) in CO 2 ; flow rate: 2.5 ml/min; wavelength: 220 nm.

Example 16: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-(1-methoxypropyl)-3,4-dihydroisoquinolin-1(2H)-one—Isomer B. ¹ H NMR (400MHz, CDCl3) δ10.88 (br s, 1H), 7.53 (s, 1H), 5.93 (s, 1H), 4.77 (s, 2H), 4.69-4.66 (m, 1H), 3.67-3.63 (m, 2H), 3.24 (s, 3H), 2.93 ( t, J=5.8Hz, 2H), 2.36 (s, 3H), 2.28 (s, 3H), 1.78-1.73 (m, 1H), 1.65-1.58 (m, 1H), 0.96 (t, J=7.2Hz, 3H). MS: 423[M+H] ⁺ . Chiral analysis: 99.6% ee; column: Chiralpak AD-H 250×4.6 mm ID, 5 μm; retention time: 8.34 minutes; mobile phase: 5% to 40% methanol (0.05% DEA) in CO 2 ; flow rate: 2.5 ml/min; wavelength: 220 nm.

Method H

Example 75: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1-[1-(hydroxyacetyl)azetidin-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one.

A solution of triphenylphosphine (15.83 g, 60.35 mmol) in anhydrous dichloromethane (16 mL) was cooled to 0°C and degassed with nitrogen for 5 minutes. Carbon tetrabromide (9.98 g, 30.1 mmol) was added and the solution was stirred at 0°C for 5 minutes. Then, it was added dropwise to a solution of tert-butyl 3-oxo-azetidine-1-carboxylate (2.52 g, 14.7 mmol) in anhydrous dichloromethane (7 mL) via syringe over 1 minute. Stirred at 0°C for 20 minutes, the reaction mixture was stirred at room temperature for 22.5 hours. Heptane (100 mL) was added and filtered to remove the resulting precipitate. The filtrate was concentrated to give an off-white solid (7.82 g). This solid was stirred in heptane (100 mL) with ultrasonic treatment, and then vacuum filtered to remove the remaining solid. The filtrate was concentrated to give tert-butyl 3-(dibromovinyl)azetidine-1-carboxylate (75a, 4.59 g, 95% yield) as a white solid. ¹ H NMR (400 MHz, CDCl 3 ) δ 4.32 (s, 4H), 1.46 (s, 9H). MS: 226, 228, 230 [M-Boc+H] ⁺ .

n-Butyllithium (1.6 M in hexanes, 1.86 mL, 2.98 mmol) was added to a solution of tert-butyl 3-(dibromovinyl)azetidine-1-carboxylate (75a, 542 mg, 1.66 mmol) in THF (16.6 mL) at -78°C. After 30 minutes, the reaction was treated with iodomethane (325 μL, 5.22 mmol) and stirred at -78°C for 1 hour. The reaction was quenched with saturated aqueous _NH4Cl and extracted with MTBE. The organic layer was concentrated and purified by silica gel chromatography (eluting with 0 to 25% ethyl acetate in heptane) to provide tert-butyl 3-(1-bromoethylidene)azetidine-1-carboxylate (75b, 0.230 g, 53% yield) as a clear oil. ¹ H NMR (400MHz, CDCl3) δ 4.38-4.43 (m, 2H), 4.31-4.37 (m, 2H), 2.14 (quint, J = 1.74Hz, 3H), 1.46 (s, 9H).

Isopropylmagnesium chloride-lithium chloride complex (1.3 M in THF, 2.00 mL, 2.60 mmol, 2.00 mL) was added to a cooled (-40°C, acetonitrile/dry ice bath) solution of 2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd S, 657 mg, 1.26 mmol) in THF (12.6 mL). The mixture was stirred for 1 hour. Tri-n-butyltin chloride (600 μL, 72.2 mmol) was added, and the flask was warmed to 0°C in an ice bath and maintained for 30 minutes. The reaction was quenched with saturated aqueous _NH₄Cl and extracted with MTBE. The MTBE layer was washed with brine and concentrated, and the resulting crude oil was purified by silica gel chromatography (eluting with 0 to 30% ethyl acetate in heptane) to afford 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(tributylstannyl)-3,4-dihydroisoquinolin-1(2H)-one (75c, 0.699 g, 76%) as a clear, viscous oil. ¹ H NMR (400MHz, DMSO-d ₆ )δ7.42 (d, J=7.09Hz, 2H), 7.37 (s, 1H), 7.22-7.34 (m, 3H), 6.74 (s, 1H), 5.37 (s, 2H), 4.69 (s, 2H), 3.22 (t, J=5.99Hz, 2H), 2.72 (t, J=5.87Hz, 2H), 2.35 (s, 3H), 2.30 (s, 3H), 1.51 (quint, J=7.76Hz, 6H), 1.08-1.35 (m, 12H), 0.85 (t, J=7.34Hz, 9H). MS: 731[M+H] ⁺ =731.

A mixture of 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(tributylstannyl)-3,4-dihydroisoquinolin-1(2H)-one (75c, 251 mg, 0.344 mmol), tert-butyl 3-(1-bromoethylidene)azetidine-1-carboxylate (75b, 103 mg, 0.393 mmol) and 1,4-dioxane (4.00 mL) was treated with tetrakistriphenylphosphine palladium(0) (60.8 mg, 0.0526 mmol) and copper(I) iodide (10.0 mg, 0.0525 mmol). The mixture was purged with nitrogen for 10 minutes, then the vial was sealed and irradiated in a microwave reactor at 120°C for 2 hours. The dioxane was removed in vacuo and the resulting oil was purified by silica gel chromatography (eluting with 0 to 40% ethyl acetate in heptane) to provide tert-butyl 3-(1-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethylidene)azetidine-1-carboxylate (75d, 49 mg, 23% yield) as a clear gum. ¹ H NMR (400MHz, CDCl3) δ7.43-7.49 (m, 2H), 7.30-7.39 (m, 3H), 7.21 (s, 1H), 6.65 (s, 1H), 5.47 (s, 2H), 4.87 (s, 2H), 4.57 (br.s. , 2H), 4.24 (br.s., 2H), 3.29 (t, J=6.24Hz, 2H), 2.73 (t, J=6.24Hz, 2H), 2.45 (s, 3H), 2.36 (s, 3H), 1.87 (s, 3H), 1.45 (s, 9H). MS: 622,624[M+H] ⁺ .

A solution of tert-butyl 3-(1-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethylidene)azetidine-1-carboxylate (75d, 49 mg, 0.079 mmol) in trifluoroacetic acid (5 mL, 70 mmol) was stirred at room temperature for 6 hours. The solution was concentrated to dryness. The residue was dissolved in methanol and purified by SCX column (Varian Bond elute SCX, 2 g, eluting with 100% MeOH to 3.5 M NH in _MeOH ) to afford crude 7-(1-(azetidin-3-yl)ethyl)-5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (Example 74, 38 mg, 100% yield) as a clear colloidal product, which was used in the next step without further purification. MS: 432, 434 [M+H] ⁺ .

Triethylamine (10 μL, 0.072 mmol) and acetoxyacetyl chloride (6.5 μL, 0.060 mmol) were added sequentially to a cooled (0°C) solution of crude 7-(1-(azetidin-3-yl)ethyl)-5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (Example 74, 24 mg, 0.056 mmol) in dichloromethane (3.0 mL). The mixture was stirred for 30 minutes and then quenched with methanol. The mixture was concentrated, dissolved in methanol (5 mL), treated with cesium carbonate (45 mg, 0.14 mmol), and stirred at room temperature overnight. The resulting solution was concentrated to dryness. The residue was dissolved in DMF, filtered, and purified by preparative HPLC to give a white powder 5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{1-[1-(hydroxyacetyl)azetidin-3-yl]ethyl}-3,4-dihydroisoquinolin-1(2H)-one (Example 75, 9.38 mg, 34% yield). ¹ H NMR (400MHz, DMSO-d ₆ )δ11.54(br.s., 1H), 7.51(s, 1H), 5.89(s, 1H), 4.81-4.99(m, 2H), 4.51-4.60(m, 3H), 4.48(br.s., 1H), 4.19(br.s. , 1H), 3.83-4.01 (m, 2H), 3.46 (t, J=6.11Hz, 2H), 2.88 (t, J=5.87Hz, 2H), 2.17 (s, 3H), 2.12 (s, 3H), 1.85 (br.s., 3H). MS; 490, 492 [M+H] ⁺ .

Method I

Example 34: (±)-5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{[1-(hydroxyacetyl)piperidin-4-yl](methyloxy)methyl}-3,4-dihydroisoquinolin-1(2H)-one.

Example 35: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{[1-(hydroxyacetyl)piperidin-4-yl](methyloxy)methyl}-3,4-dihydroisoquinolin-1(2H)-one—Isomer B.

Example 36: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{[1-(hydroxyacetyl)piperidin-4-yl](methyloxy)methyl}-3,4-dihydroisoquinolin-1(2H)-one—Isomer A.

Isopropylmagnesium chloride-lithium chloride complex (1.3 M in THF, 0.850 mL, 1.10 mmol) was added to a -40°C solution of 2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd S, 311.0 mg, 0.598 mmol) in tetrahydrofuran (5.0 mL) and 1,4-dioxane (0.5 mL). The mixture was stirred for 1 hour. N-Boc-4-formylpiperidine (0.242 g, 1.14 mmol) was then added, and the flask was warmed to 0°C in an ice bath. After 1 hour at 0°C, the reaction was quenched with saturated aqueous NH ₄ Cl and extracted with MTBE. The MTBE layer was concentrated and the resulting oil was purified on silica gel (Isco RediSepRf, 12 g, gradient 10 to 70% ethyl acetate in heptane) to afford racemic tert-butyl 4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)(hydroxy)methyl)piperidine-1-carboxylate ( ^8a , 0.229 g, 59%) as a white solid. NMR (400MHz, CD3OD) δ7.66 (s, 1H), 7.36-7.44 (m, 2H), 7.18-7.31 (m, 3H), 6.7 1 (s, 1H), 5.42 (s, 2H), 5.04 (d, J = 5.14Hz, 1H), 4.83 (d, J = 1.96Hz, 2H), 4.08 ( d, J=12.72Hz, 2H), 3.23 (t, J=6.24Hz, 2H), 2.73 (t, J=6.11Hz, 2H), 2.53-2.7 1(m, 2H), 2.39(s, 3H), 2.35(s, 3H), 1.76-1.90(m, 1H), 1.32-1.62(m, 13H); MS 654, 656 [M+H] ⁺ .

A cooled (0° C.) solution of racemic tert-butyl 4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)(hydroxy)methyl)piperidine-1-carboxylate (8a, 91.0 mg, 0.139 mmol) in THF (3.0 mL) was treated with iodomethane (34 mg, 0.24 mmol) and potassium tert-butoxide (1.0 M in THF, 0.155 mL, 0.155 mmol). Stirring was continued at 0° C. for 30 minutes, and the mixture was extracted by partitioning between brine and MTBE. The organic phase was concentrated to dryness to give colloidal crude racemic tert-butyl 4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)(methoxy)methyl)piperidine-1-carboxylate (34b, 97 mg, 100%). MS: 612, 614 [M+H-tBu] ⁺ .

Trifluoroacetic acid (0.10 mL, 1.35 mmol) was added to a room temperature solution of crude racemic tert-butyl 4-((2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)(methoxy)methyl)piperidine-1-carboxylate (34b, 97 mg, 0.139 mmol) in dichloromethane (5.0 mL). The mixture was stirred at room temperature for 2 hours, at 35° C. for 4 hours, at room temperature overnight, and then at 40° C. for 6 hours. The solution was diluted with heptane and concentrated to dryness to give crude racemic 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(methoxy(piperidin-4-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (34c, 124 mg, 100%) as a gel. MS: 568, 570 [M+H] ⁺ .

Triethylamine (75 μL, 0.54 mmol) and 2-acetoxyacetyl chloride (16 μL, 0.15 mmol) were added to a cooled (0°C) solution of crude racemic 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(methoxy(piperidin-4-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (34c, 124 mg, 0.139 mmol) in dichloromethane (3.0 mL). The mixture was stirred at 0°C for 30 minutes. Trifluoroacetic acid (2.0 mL) was then added, and the mixture was stirred at room temperature for 1 hour, then at 40°C for 7 hours. The solution was concentrated to dryness and dried under high vacuum for 2 days to afford crude racemic 2-(4-((5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)(methoxy)methyl)piperidin-1-yl)-2-oxoethyl acetate (34d, 204 mg, 100%) as a golden oil. MS: 578, 580 [M+H] ⁺ .

Crude racemic 2-(4-((5,8-dichloro-2-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7 _- yl)(methyloxy)methyl)piperidin-1-yl)-2-oxoethyl acetate (34d, 204 mg, 0.139 mmol) was dissolved in 7N ammonia in methanol (4 mL, 28 mmol NH 3 ) and stirred at room temperature for 1 hour and then at 40° C. for 4 hours. The solution was concentrated to dryness, and the residue was purified by preparative HPLC to give (±)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{[1-(hydroxyacetyl)piperidin-4-yl](methyloxy)methyl}-3,4-dihydroisoquinolin-1(2H)-one as a white solid (Example 34, 19.87 mg, ²⁷ % yield from 8a). NMR (400MHz, CD3OD) δ7.44 (s, 1H), 6.01 (s, 1H), 4.66 (s, 2H), 4.56 (d, J=5.62Hz, 1H), 4.35-4.45(m, 1H), 4.02-4.17(m, 2H), 3.57-3.68(m, 1H), 3.43(t, J=6.24Hz , 2H), 3.10 (s, 3H), 2.86-2.92 (m, 2H), 2.75-2.85 (m, 1H), 2.40-2.54 (m, 1H), 2.2 0(s, 3H), 2.15(s, 3H), 1.78-1.87(m, 1H), 1.60-1.69(m, 1H), 1.18-1.47(m, 3H). MS: 536, 538[M+H] ⁺ .

The racemate (Example 34) was further purified by chiral preparative SFC to afford Example 35 (Isomer B, retention time 13.019 minutes, 9.09 mg, 12% yield from 8a) and Example 36 (Isomer A, retention time 10.712 minutes, 8.36 mg, 11% yield from 8a) as white solids after freeze-drying. The absolute stereochemistry of the benzyl carbon in each isomer was not determined.

Example 35: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{[1-(hydroxyacetyl)piperidin-4-yl](methyloxy)methyl}-3,4-dihydroisoquinolin-1(2H)-one—Isomer B. MS: 536, 538 [M+H] ⁺ . Chiral analysis: ~97% ee, retention time 13.019 minutes, Lux Cellulose-4 4.6×100 mm3u column, eluted with 50% MeOH, 120 bar, 4 mL/min.

Example 36: 5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{[1-(hydroxyacetyl)piperidin-4-yl](methyloxy)methyl}-3,4-dihydroisoquinolin-1(2H)-one—Isomer A. MS: 536, 538 [M+H] ⁺ . Chiral analysis: >99% ee, retention time 10.712 minutes, on a Lux Cellulose-4 4.6×100 mm³ column, eluted with 50% MeOH, 120 bar, 4 mL/min.

Example 81: 5,8-Dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[(R)-methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one.

Example 82: 5,8-Dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[(S)-methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one.

Solid pyridinium dichromate (5.87 g, 15.6 mmol) was added in five portions to a stirred solution of oxetane-3-ylmethanol (2.20 g, 24.97 mmol) in dichloromethane (110 mL) at room temperature. The resulting black mixture was stirred at room temperature for 16 hours. The dark brown suspension was then filtered through a pad of silica gel, and the filter cake was rinsed with dichloromethane (8 x 120 mL). The combined dichloromethane filtrates were partially concentrated under reduced pressure at room temperature (27-30°C) to afford oxetane-3-carbaldehyde (81a, 3 g, ~26% yield) as a colorless solution, 18.7 wt% in dichloromethane by NMR. The solution was dried over magnesium sulfate, filtered, and used immediately in the next step. 1H NMR (400MHz, CDCl3) δ9.95 (d, J = 2.4Hz, 1H), 4.87 (m, 4H), 3.81 (m, 1H).

A solution of 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd SS, 500 mg, 0.932 mmol) in anhydrous THF (7 mL) was cooled to -65°C, followed by the dropwise addition of isopropylmagnesium chloride-lithium chloride complex (1.3 M in THF, 2.15 mL, 2.80 mmol) over 3 minutes. The resulting brown solution was stirred at -65°C for 10 minutes, then warmed to -10°C and maintained for 30 minutes. Oxetane-3-carbaldehyde (81a, ~2.2 g, ~4.8 mmol, 18.7 wt% in dichloromethane) was added dropwise over 2 minutes, causing the color to turn light yellow. Stirring was continued at -5°C for 30 minutes. The reaction was quenched with glacial acetic acid (0.5 mL), diluted with ethyl acetate (10 mL), and washed with saturated aqueous _NaHCO₃ /saturated aqueous NaCl (1/1 v/v, 3 x 15 mL). The organic layer was dried over sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (eluting with 1/1 petroleum ether/ethyl acetate) to afford racemic 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichloro-7-(hydroxy(oxetan-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (81b, 300 mg, 59% yield) as a white solid. MS: 543 [M+H] ^⁺ .

Iodomethane (133 mg, 0.938 mmol) was added dropwise to a cooled (-5°C) suspension of 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichloro-7-(hydroxy(oxetan-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (81b, 300 mg, 0.552 mmol) in anhydrous THF (5 mL). Potassium tert-butoxide (1.0 M in THF, 0.938 mL, 0.938 mmol) was added, and the mixture was stirred at 0°C for 1 hour. The reaction mixture was extracted by partitioning with saturated aqueous NaCl solution (15 mL) and MTBE (3 x 15 mL). The combined organic extracts were washed with saturated aqueous NaCl (30 mL), dried over sodium sulfate, concentrated, and purified by silica gel chromatography (eluting with 1/1 petroleum ether/ethyl acetate) to provide racemic 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichloro-7-(methoxy(oxetan-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (81c, 280 mg, 91% yield), a yellow gum. MS: 557 [M+H] ⁺ .

A mixture of 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichloro-7-(methoxy(oxetan-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (81c, 100 mg, 0.179 mmol), PtO ₂ (21 mg, 0.092 mmol) and ethyl acetate (4 mL) was stirred at room temperature under hydrogen balloon pressure for 3 days. The solution was filtered through a pad of Celite. The flask and filter pad were rinsed with ethyl acetate (2×10 mL). The combined filtrates were concentrated and purified by preparative thin layer chromatography (silica gel, eluting with 10/1 dichloromethane/methanol) to give racemic 5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one (81d, 45 mg, 54% yield) as a white solid.

Multiple batches of racemic 5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one (81d, total 140 mg) were mixed and chirally separated by preparative SFC [column: (R,R) Whelk O1 250mm*30mm, 5μm; mobile phase: base-ETOH; wavelength: 220nm; workup: freeze drying] to give Example 81 (50.34mg, 36% yield) as a gray solid, which was further purified by preparative TLC (silica gel, eluted with 10/1 dichloromethane/methanol) to give Example 82 (22.83g, 16% yield) as a brown solid. The small molecule X-ray crystal structure of Example 82 shows that it has absolute (S) stereochemistry, so its enantiomer Example 81 belongs to absolute (R) stereochemistry.

Example 81: 5,8-Dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[(R)-methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one. ¹ H NMR (400MHz, CDCl3) δ12.34 (brs, 1H), 7.49 (s, 1H), 5.93 (s, 1H), 5.05 (d, J=6.0Hz, 1H), 4.78-4.61 (m, 6 H), 3.88 (s, 3H), 3.50-3.48 (m, 2H), 3.38-3.37 (m, 1H), 3.31 (s, 3H), 2.94 (t, J=6.2Hz, 2H), 2.35 (s, 3H). MS: 489[M+Na]+. Chiral analysis: 100% ee; retention time 9.85 minutes; column (R,R) WhelkO1, 250×4.6 mm ID, 5 μm; mobile phase: 50% ethanol (0.05% DEA) in CO 2 ; wavelength 220 nm.

Example 82: 5,8-Dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[(S)-methoxy(oxetan-3-yl)methyl]-3,4-dihydroisoquinolin-1(2H)-one. ¹ H NMR (400MHz, CDCl3) δ12.38 (brs, 1H), 7.49 (s, 1H), 5.92 (s, 1H), 5.05 (d, J=6.0Hz, 1H), 4.78-4.64 (m, 6 H), 3.87 (s, 3H), 3.50-3.47 (m, 2H), 3.38-3.37 (m, 1H), 3.31 (s, 3H), 2.93 (t, J=6.2Hz, 2H), 2.35 (s, 3H). MS: 467[M+H] ⁺ . Chiral analysis: 98% ee; retention time 8.65 minutes; column: (R,R) WhelkO1, 250×4.6 mm ID, 5 μm; mobile phase: 50% ethanol (0.05% DEA) in CO 2 ; wavelength 220 nm.

Example 83: 8-Chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(R)-methoxy[(3R)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinolin-1(2H)-one.

Example 84: 8-Chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(R ^* )-methoxy[(3S ^* )-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinolin-1(2H)-one.

Example 85: 8-Chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(S ^* )-methoxy[(3R ^* )-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinolin-1(2H)-one.

Example 86: 8-Chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(S)-methoxy[(3S)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinolin-1(2H)-one.

An aqueous solution of tetrahydrofuran-3-carbaldehyde (~4.0 mL of a 50 wt% aqueous solution, 4.0 g) was extracted with dichloromethane (2 x 2.5 mL). The combined organic layers were cooled (15° C.) and phosphorus pentoxide was slowly added. The resulting dark solution was filtered to remove solids, and the yellow filtrate (~16 wt% tetrahydrofuran-3-carbaldehyde in dichloromethane by NMR) was quickly used as described below.

Isopropylmagnesium chloride-lithium chloride complex (1.3 M in THF, 3.73 mL, 4.85 mmol) was added dropwise over 5 minutes to a cooled (-70°C) solution of 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-iodo-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Cpd UU, 910 mg, 1.617 mmol) in anhydrous THF (10 mL). The resulting brown mixture was stirred at -70°C for 30 minutes, followed by the dropwise addition of the tetrahydrofuran-3-carbaldehyde solution prepared above (3.98 g of a ~16 wt% solution in dichloromethane, ~636 mg of tetrahydrofuran-3-carbaldehyde, ~16.35 mmol) over 5 minutes. Stirring was continued at -70°C for 30 minutes. The reaction was quenched with glacial acetic acid (0.5 mL), diluted with ethyl acetate (80 mL), and then washed with saturated aqueous NaHCO ₃ solution/saturated aqueous NaCl solution (1/1 v/v, 3×35 mL). The organic layer was dried over sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (eluting with 1/1 petroleum ether/ethyl acetate) to afford 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-(hydroxy(tetrahydrofuran-3-yl)methyl)-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (83a, 450 mg, 52% yield, mixture of 4 diastereomers) as a white solid. ¹ H NMR (400MHz, CDCl3) δ7.37-7.46 (m, 3H), 7.20-7.31 (m, 3H), 6.39 (s, 1H), 5.44 (s, 2H), 5.22-5.31 (m, 1H), 4.87 (s, 2H), 3.88-4.02 (m, 1H), 3.8 3(s, 3H), 3.65-3.82(m, 3H), 3.16(t, J=5.99Hz, 2H), 2.71-2.89(m, 1H) , 2.55 (t, J=6.17Hz, 2H), 2.44 (s, 3H), 2.20 (s, 3H), 1.66-1.95 (m, 3H).

To a cooled (-5°C) suspension of 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-(hydroxy(tetrahydrofuran-3-yl)methyl)-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (83a, 500 mg, 0.931 mmol) in THF (15 mL) was added iodomethane (225 mg, 1.58 mmol) dropwise, followed by the addition of potassium tert-butoxide (1.0 M in THF, 1.58 mL, 1.58 mmol). The mixture was stirred at 0°C for 1 hour. Glacial acetic acid (0.5 mL) and ethyl acetate (100 mL) were added, and the solution was washed with saturated aqueous NaHCO₃ ( ₃ x 20 mL) and brine (30 mL). The organic layer was dried over sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (eluting with 1/1 petroleum ether/ethyl acetate) to give 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-(methoxy(tetrahydrofuran-3-yl)methyl)-5-methyl-3,4-dihydroisoquinolin-1(2H)-one as a mixture of 4 diastereomers (400 mg, 70% yield).

The stereoisomers were separated by preparative chiral SFC (column: AD, 250*30 mm, 5 μm, mobile phase: 30% IPA + NH ₃ H ₂ O, 60 ml/min, wavelength: 220 nm, treatment: freeze drying) to obtain 140 mg of white solid peak 12 (a mixture of peaks 1 and 2) and 120 mg of white solid peak 34 (a mixture of peaks 3 and 4).

The peak 12 mixture (140 mg) was separated by preparative SFC (column: AD, 250*30 mm, 5 μm, mobile phase: 25% MeOH+ _{NH3H2O 60} ml/min, wavelength: 220 nm, workup: freeze drying) to give enantiomerically enriched peak 1 (60 mg, 81% chiral purity, further purified as described below) and pure peak 2 (1610b, 60 mg, 12% yield, 96% chiral purity, the compound was used without further purification).

The peak 34 mixture (120 mg) was separated by preparative SFC (column: AD, 250*30 mm, 5 μm, mobile phase: 25% MeOH+ _{NH3H2O 60} ml/min, wavelength: 220 nm, workup: freeze drying) to give pure peak 3 (1613b, 50 mg, 9.8% yield, 95% chiral purity, the compound was used without further purification) and enantiomerically enriched peak 4 (50 mg, 88% chiral purity, further purified as described below).

The enantiomerically enriched peak 1 material (60 mg, 81% chiral purity) was further purified by preparative SFC (column: AD, 250*30 mm, 5 μm, mobile phase: 25% MeOH+ _{NH3H2O 70} mL/min, wavelength: 220 nm, workup: freeze drying) to give pure peak 1 (83b, 45 mg, 8.7% yield, 98% chiral purity).

The enantiomerically enriched peak 4 material (50 mg, 88% chiral purity) was further purified by preparative SFC (column: AD, 250*30 mm, 5 μm, mobile phase: 25% MeOH + NH ₃ H ₂ O 70 ml/min, wavelength: 220 nm, workup: freeze drying) to give pure peak 4 (1613b, 45 mg, 8.7% yield, 99% chiral purity). The absolute or relative stereochemistry of the individual isomers was not determined at this stage.

A stirred solution of 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-8-chloro-7-(methoxy(tetrahydrofuran-3-yl)methyl)-5-methyl-3,4-dihydroisoquinolin-1(2H)-one peak 1 (83b, 45 mg, 0.082 mmol) in dichloromethane (2 mL) was treated with TFA (2 mL) at room temperature. The mixture was heated to 30° C. for 16 hours, then the solution was diluted with dichloromethane (20 mL) and concentrated to dryness. The residue was purified by silica gel chromatography (eluting with 10/1 dichloromethane/methanol) to afford 8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(R)-methoxy[(3R)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinolin-1(2H)-one as a white solid (Example 83, 18.55 mg, 50% yield, 100% ee). The small molecule X-ray crystal structure of Example 83 confirmed its absolute (R,R) stereochemistry. ¹ H NMR (400MHz, CDCl3) δ12.36 (brs, 1H), 7.29 (s, 1H), 5.91 (s, 1H), 4.85-4.75 (m, 3H), 3.89-3.83 (m, 6H), 3.71-3.69 (m, 1 H), 3.47-3.44(m, 2H), 3.16(s, 3H), 2.76-2.73(m, 2H), 2.58-2.54(m, 1H), 2.34(s, 3H), 2.26(s, 3H), 1.73-1.70(m, 2H). MS: 461[M+H] ⁺ . Chiral analysis: 100% ee; retention time 34.91 minutes; column: Chiralpak IC 250×4.6 mm ID, 5 μm; mobile phase: 50% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.0 ml/min; wavelength: 220 nm.

Peak 2 stereoisomer (1610b, 60.0 mg, 0.109 mmol) was obtained as a white solid, 8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(R ^* )-methoxy[(3S ^* )-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Example 84, 30 mg, 60% yield, 97% ee). The absolute stereochemistry of this isomer was not determined, but the ¹ H NMR spectrum showed that it was significantly different from that of Example 83, indicating that Example 84 was an R,S or S,R isomer. ¹ H NMR (400MHz, CDCl3) δ12.33 (br.s, 1H), 7.30 (s, 1H), 5.91 (s, 1H), 4.84-4.77 (m, 3H), 3.89-3.86 (m, 4H), 3.73-3.68 (m, 2H), 3.60- 3.58 (m, 1H), 3.46-3.44 (m, 2H), 3.19 (s, 3H), 2.75-2.73 (m, 2H), 2.64-2.62 (m, 1H), 2.34 (s, 3H), 2.24 (s, 3H), 1.98-1.95 (m, 2H). MS: 461[M+H] ⁺ . Chiral analysis: 97% ee, retention time 39.01 minutes; column: Chiralpak IC 250×4.6 mm ID, 5 μm; mobile phase: 50% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.0 ml/min; wavelength: 220 nm.

Peak 3 stereoisomer (85b, 50.0 mg, 0.091 mmol) was obtained as a white solid, 8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(S ^* )-methoxy[(3R ^* )-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Example 85, 16.06 mg, 38% yield, 100% ee). The absolute stereochemistry of this isomer was not determined, but the ¹ H NMR spectrum showed that it was significantly different from that of Example 83 and was the same as that of Example 84, indicating that Example 85 was an S,R or R,S isomer. ¹ H NMR (400MHz, CDCl3) δ12.30 (br s, 1H), 7.30 (s, 1H), 5.91 (s, 1H), 4.84-4.77 (m, 3H), 3.89-3.86 (m, 4H), 3.74-3.66 (m, 2H), 3.59-3.58 (m, 1H), 3. 46-3.44(m, 2H), 3.19(s, 3H), 2.75-2.73(m, 2H), 2.65-2.63(m, 1H), 2.34(s, 3H), 2.24(s, 3H), 1.98-1.95(m, 2H). MS: 461[M+H] ⁺ . Chiral analysis: 100% ee, retention time 29.05 minutes; column: Chiralpak IC 250×4.6 mm ID, 5 μm; mobile phase: 50% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.0 ml/min; wavelength: 220 nm.

Peak 4 stereoisomer (86b, 45.0 mg, 0.082 mmol) was obtained as a white solid, 8-chloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-{(S)-methoxy[(3S)-tetrahydrofuran-3-yl]methyl}-5-methyl-3,4-dihydroisoquinolin-1(2H)-one (Example 86, 14.79 mg, 39% yield, 100% ee). Although it had a different retention time on a chiral column, the ¹ H NMR spectrum of this compound was identical to that of Example 83, and X-ray crystallography revealed that Example 83 had R,R stereochemistry. This indicated that Example 86 was the S,S stereoisomer. ¹ H NMR (400MHz, CDCl3) δ12.29 (br.s, 1H), 7.30 (s, 1H), 5.91 (s, 1H), 4.84-4.75 (m, 3H), 3.89-3.83 (m, 6H), 3.71-3.69 (m, 1 H), 3.47-3.44(m, 2H), 3.16(s, 3H), 2.76-2.73(m, 2H), 2.58-2.56(m, 1H), 2.34(s, 3H), 2.25(s, 3H), 1.73-1.70(m, 2H). MS: 461[M+H] ⁺ . Chiral analysis: 100% ee, retention time 32.28 minutes; column: Chiralpak IC 250×4.6 mm ID, 5 μm; mobile phase: 50% ethanol (0.05% DEA) in CO 2 ; flow rate: 2.0 ml/min; wavelength: 220 nm.

Method J

Example 87: 5,8-Dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[1-(1-methylazetidin-3-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one—Isomer A.

Example 88: 5,8-Dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[1-(1-methylazetidin-3-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one—Isomer B.

A solution of 1-boc-azetidine-3-carboxylic acid (5.00 g, 24.8 mmol) and CDI (4.23 g, 26.1 mmol) in dichloromethane (100 mL) was stirred at room temperature for 1 hour. N,O-Dimethylhydroxylamine hydrochloride (4.0 g, 29.8 mmol) was then added and stirring continued at room temperature for 16 hours. The resulting suspension was washed with water (3 x 30 mL), saturated aqueous _NaHCO₃ (3 x 30 mL), and brine (3 x 30 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to afford tert-butyl 3-(methoxy(methyl)carbamoyl)azetidine-1-carboxylate (87a, 5.1 g, 84% yield) as a colorless oil. ¹ H NMR (400MHz, CDCl3) δ4.14 (br s, 2H), 4.05 (t, J = 8.6Hz, 2H), 3.66 (s, 3H), 3.65 (m, 1H), 3.20 (s, 3H), 1.43 (s, 9H).

Methylmagnesium bromide (3 M in THF, 10.4 mL, 31.3 mmol) was added dropwise to a cooled (0°C) solution of tert-butyl 3-(methoxy(methyl)carbamoyl)azetidine-1-carboxylate (87a, 5.1 g, 20.88 mmol) in anhydrous THF (100 mL). Stirring was continued at 0°C for 1 hour, followed by 16 hours at room temperature. The mixture was cooled to 0°C, quenched with saturated aqueous _NaHCO₃ (35 mL), and extracted with ethyl acetate (3 x 40 mL). The combined organic extracts were washed with brine (3 x 40 mL), dried over sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (eluting with 10:1 to 3:1 petroleum ether/ethyl acetate) to afford tert-butyl 3-acetylazetidine-1-carboxylate (87b, 3.20 g, 77% yield) as a light yellow oil. ¹ HNMR (400MHz, CDCl3) δ4.05 (d, J=7.6Hz, 4H), 3.41 (quint, J=7.6Hz, 1H), 2.18 (s, 3H), 1.43 (s, 9H).

A solution of 2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd SS, 1.20 g, 2.238 mmol) in anhydrous THF (15 mL) was cooled to -60°C, followed by the addition of isopropylmagnesium chloride-lithium chloride complex (1.3 M in THF, 5.16 mL, 6.71 mmol) dropwise via syringe over 3 minutes. Stirring was continued at -60°C for 10 minutes, followed by stirring at 0°C for 20 minutes. tert-Butyl 3-(methoxy(methyl)carbamoyl)azetidine-1-carboxylate (87a, 892 mg, 4.48 mmol) was added dropwise, and the mixture was stirred at 0°C for 1 hour. The reaction was quenched with glacial acetic acid (1 mL) and diluted with ethyl acetate (100 mL). The organic phase was washed with NaHCO ₃ /brine (v/v = 1/1, 3×50 mL) and brine (50 mL), dried over sodium sulfate, and filtered. The filtrate was concentrated to give a crude yellow oil (2.0 g). Purification by silica gel chromatography (eluting with petroleum ether/ethyl acetate = 1:1) afforded racemic tert-butyl 3-(1-(2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-1-hydroxyethyl)azetidine-1-carboxylate (87c, 450 mg, 31% yield) as a white solid. MS: 656 [M+H] ⁺ .

Trifluoromethanesulfonic acid (0.54 mL, 6.15 mmol) was added dropwise to a cooled (0°C) solution of racemic tert-butyl 3-(1-(2-((2-(benzyloxy)-4-methoxy-6-methylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)-1-hydroxyethyl)azetidine-1-carboxylate (87c, 450 mg, 0.685 mmol) in anhydrous dichloromethane (10 mL). The mixture was stirred at 5-10°C for 1 hour, then cooled to 0°C and trifluoromethanesulfonic acid (0.54 mL, 6.15 mmol) was added. After stirring at 2-5°C for 12 hours, saturated aqueous sodium bicarbonate was added to bring the pH to ~8. The mixture was concentrated to remove dichloromethane, and the aqueous residue was diluted with THF (20 mL). Solid sodium bicarbonate (288 mg, 3.43 mmol) and di-tert-butyl dicarbonate (448 mg, 2.06 mmol) were added and the mixture was stirred at 2-5°C for 16 hours and then at 15°C for 18 hours. The solution was extracted with ethyl acetate (2 x 50 mL). The combined organic layers were washed with brine (2 x 20 mL), dried over sodium sulfate, and filtered. The filtrate was concentrated to give a crude yellow solid (1 g), which was first purified by silica gel chromatography (eluting with 10% methanol/dichloromethane) and then by preparative SFC (column: AD 250 mm*30 mm, 5 μm; mobile phase: 35% Base-ETOH; wavelength: 220 nm; workup: concentration) to afford tert-butyl 3-(1-(5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)vinyl)azetidine-1-carboxylate (87d, 256 mg, 68.1%) as a yellow solid. MS: 548 [M+H] ⁺ .

A suspension of tert-butyl 3-(1-(5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)vinyl)azetidine-1-carboxylate (87d, 236 mg, 0.43 mmol) and platinum oxide (80 mg, 0.35 mmol) in ethyl acetate (15 mL) and methanol (5 mL) was stirred at room temperature under hydrogen balloon pressure for 3 hours. The solid was removed by filtration, and the filtrate was concentrated and purified by preparative TLC (silica gel, eluted with dichloromethane/methanol = 15:1) to give racemic tert-butyl 3-(1-(5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl)azetidine-1-carboxylate (87e, 180 mg, 76% yield) as a white solid. MS: 549 [M+H] ⁺ .

A solution of the resulting racemic tert-butyl 3-(1-(5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl)azetidine-1-carboxylate (87e, 180 mg, 0.327 mmol) in dichloromethane (10 mL) was stirred with HCl (4.0 M in methanol, 5 mL, 20 mmol) at 14° C. for 30 minutes. The solution was concentrated to dryness, and the residue was dissolved in methanol (5 mL). Concentrated _NH4OH was added to bring the pH to ~8 and the mixture was concentrated to dryness to give crude racemic 7-(1-(azetidin-3-yl)ethyl)-5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (87f, 180 mg, 100%) as a white solid which was used without further purification.

Glacial acetic acid (0.1 mL) was added to a 15°C solution of crude racemic 7-(1-(azetidin-3-yl)ethyl)-5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3,4-dihydroisoquinolin-1(2H)-one (87f, 180 mg, 0.327 mmol) and formaldehyde (37 wt% aqueous solution, 79.5 mg, 0.980 mmol) in methanol (5 mL). The mixture was stirred at 15°C for 45 minutes. Sodium cyanoborohydride (41 mg, 0.653 mmol) was added, and stirring was continued at room temperature for 12 hours. The reaction was quenched with saturated _NH4Cl solution (2 mL), stirred at 15°C for 30 minutes, and then concentrated to remove the solvent. The residue was dissolved in dichloromethane/methanol (v/v = 10:1, 50 mL) and filtered. The filtrate was concentrated and purified by preparative HPLC [column: Phenomenex Gemini C18 250*50 10μ; mobile phase: 4% to 34% acetonitrile (containing 0.225% formic acid)/water; wavelength: 220 nm; workup: freeze drying] to give the formate salt of (±)-5,8-dichloro-2-((4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-7-(1-(1-methylazetidin-3-yl)ethyl)-3,4-dihydroisoquinolin-1(2H)-one as a white solid (110 mg, 66%). The enantiomers of this racemic salt were separated by preparative SFC [column: AD (250 mm*30 mm, 5 μm); mobile phase: 30% base-ETOH; wavelength: 220 nm; workup: freeze-drying], and each enantiomer was purified separately by preparative HPLC [column: Phenomenex Gemini C18 250*50 10 μm; mobile phase: 28% MeCN (containing 0.05% ammonia)/water to 48% MeCN (containing 0.05% ammonia)/water; wavelength: 220 nm; workup: freeze-drying] to provide white solids Isomer A (Example 87, 15.67 mg, 16% yield) and Isomer B (Example 88, 13.35 mg, 13% yield). The absolute stereochemistry of each isomer was not determined.

Example 87: 5,8-Dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[1-(1-methylazetidin-3-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one—Isomer A. ¹ H NMR (400MHz, CD3OD) δ7.44 (s, 1H), 6.27 (s, 1H), 4.73 (s, 2H), 3.91 (m, 3H), 3.77-3.68 (m, 2H), 3.41-3.37 (m, 3H) , 3.16 (t, J=7.4Hz, 1H), 2.95-2.92 (m, 2H), 2.90-2.83 (m, 2H), 2.39 (s, 3H), 2.34 (s, 3H), 1.16 (d, J=6.4Hz, 3H). MS: 464[M+H] ⁺ . Chiral analysis: 99% ee; retention time 5.511 minutes, Chiralpak AD-3 150×4.6 mm ID, 3 μm column [mobile phase: A: CO2 B: ethanol (0.05% DEA); gradient: 5% to 40% B in 5.0 minutes and hold at 40% for 2.5 minutes, followed by 5% B for 2.5 minutes; wavelength: 220 nm].

Example 88: 5,8-Dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[1-(1-methylazetidin-3-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one—Isomer B. ¹ H NMR (400MHz, CD3OD) δ7.43 (s, 1H), 6.26 (s, 1H), 4.73 (s, 2H), 3.91 (m, 3H), 3.77-3.68 (m, 2H), 3.41-3.37 (m, 3H), 3.12(brs, 1H), 2.95-2.92(m, 2H), 2.90-2.83(m, 2H), 2.36(s, 3H), 2.33(s, 3H), 1.16(d, J=6.4Hz, 3H). MS: 464[M+H] ⁺ . Chiral analysis: 100% ee; retention time 5.997 minutes, Chiralpak AD-3 150×4.6 mm ID, 3 μm column [mobile phase: A: CO 2 B: ethanol (0.05% DEA); gradient: 5% to 40% B in 5.0 minutes and hold at 40% for 2.5 minutes, followed by 5% B for 2.5 minutes; wavelength: 220 nm].

Method K

Example 89: (±)-5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[1-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one.

Example 90: (+)-5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[1-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one.

Example 91: (-)-5,8-Dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[1-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one.

Isopropylmagnesium chloride-lithium chloride complex (1.3 M in THF, 22.2 mL, 28.8 mmol) was added via syringe to a cooled (-40°C) solution of 2-{[2-(benzyloxy)-4,6-dimethylpyridin-3-yl]methyl}-7-bromo-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (Cpd S, 5.00 g, 9.61 mmol) in anhydrous THF (50 mL) and 1,4-dioxane (5 mL). The mixture was stirred at -40°C for 30 minutes, and zinc chloride (1.0 M in diethyl ether, 11.5 mL, 11.5 mmol) was added. Stirring was continued at -40°C for 30 minutes, followed by the addition of tetrakis(triphenylphosphine)palladium(0) (1.11 g, 0.961 mmol) and acetyl chloride (1.51 g, 19.2 mmol). The mixture was stirred for 18 hours and allowed to warm to room temperature. The reaction was quenched with saturated aqueous _NH4Cl (5 mL), diluted with ethyl acetate (30 mL), and washed with saturated aqueous _NH4Cl (40 mL) and brine (40 mL). The organic layer was dried over sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (eluting with 0 to 30% ethyl acetate in petroleum ether) to provide 7-acetyl-2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (89a, 4.00 g, 78% yield) as a yellow solid. ^1H NMR (400MHz, CDCl3) δ7.45 (d, J=6.8Hz, 2H), 7.39 (s, 1H), 7.38-7.27 (m, 3H), 6.63 (s, 1H), 5.43 (s, 2H) ), 4.86 (s, 2H), 3.29 (t, J=6.3Hz, 2H), 2.75 (t, J=6.3Hz, 2H), 2.63 (s, 3H), 2.42 (s, 3H), 2.34 (s, 3H). MS: 483[M+H] ⁺ .

Sodium borohydride (47.0 mg, 1.24 mmol) was added to a room temperature solution of 7-acetyl-2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-3,4-dihydroisoquinolin-1(2H)-one (89a, 200 mg, 0.414 mmol) in methanol (5 mL). The mixture was stirred at room temperature for 30 minutes, and the reaction mixture was concentrated and purified by silica gel chromatography (eluting with 0 to 30% ethyl acetate in petroleum ether) to provide 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-hydroxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (89b, 190 mg, 95% yield) as a white solid.

A cooled (0°C) solution of 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-hydroxyethyl)-3,4-dihydroisoquinolin-1(2H)-one (89b, 190 mg, 0.391 mmol) and triethylamine (119 mg, 1.17 mmol) in dichloromethane (5 mL) was treated with methanesulfonyl chloride (67.3 mg, 0.587 mmol) and stirred at 0°C for 1 hour. The reaction mixture was diluted with dichloromethane (30 mL); washed sequentially with saturated aqueous _NH4Cl solution, saturated aqueous _NaHCO3 solution, and saturated aqueous NaCl solution; dried over sodium sulfate, and concentrated to dryness to afford crude racemic 1-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl methanesulfonate (89c, 225 mg, 100% yield) as a white solid, which was used immediately without further purification.

A suspension of 1-(2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)ethyl methanesulfonate (89c, 100 mg, 0.177 mmol), morpholine (46.4 mg, 0.532 mmol), and potassium carbonate (73.6 mg, 0.532 mmol) in acetonitrile (5 mL) was stirred at reflux (85° C.) for 2 hours. After cooling to room temperature, the suspension was filtered to remove the solid. The filtrate was concentrated and purified by silica gel chromatography (eluting with 0 to 30% ethyl acetate/petroleum ether) to provide colloidal racemic 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-morpholinoethyl)-3,4-dihydroisoquinolin-1(2H)-one (89d, 90 mg, 91% yield, 80% purity by LCMS). MS: 576 [M+Na] ⁺ .

A solution of racemic 2-((2-(benzyloxy)-4,6-dimethylpyridin-3-yl)methyl)-5,8-dichloro-7-(1-morpholinoethyl)-3,4-dihydroisoquinolin-1(2H)-one (89d, 90 mg, 0.16 mmol) in dichloromethane (5 mL) and trifluoroacetic acid (2 mL) was stirred at room temperature for 19 hours. The solution was evaporated to dryness, and the residue was dissolved in toluene (10 mL) and basified to pH 8 to 9 by adding a few drops of concentrated _NH4OH . The solution was concentrated and purified by preparative HPLC to give (±)-5,8-dichloro-2-[(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[1-(morpholin-4-yl)ethyl]-3,4-dihydroisoquinolin-1(2H)-one (Example 89, 29.49 mg, 39% yield) as a white solid. ¹ H NMR (400MHz, CDCl3) δ7.74 (s, 1H), 5.94 (s, 1H), 4.87-4.70 (m, 2H), 4.00 (q, J=6.3Hz, 1H), 3.76-3 .59 (m, 6H), 2.99-2.81 (m, 2H), 2.53 (br.s., 2H), 2.36 (m, 5H), 2.29 (s, 3H), 1.24 (d, J=6.5Hz, 3H). MS: 464[M+H] ⁺ .

The racemic material (Example 89) was further purified by chiral SFC separation conditions to afford the compound of Example 90 and the compound of Example 91.

Additional compounds of the invention were prepared by variations of the methods exemplified in this application. Selected compounds prepared are presented in Table 1 below along with corresponding characterization data.

Table 1

Biological experiments and data

Purification of WT and mutant EZH2Y641N

WT and mutant EZH2 were purified using the same procedure. Genes for EZH2, EED, SUZ12, and RBBP4 proteins were cloned into the pBacPAK9 vector (Clontech). RBBP4 was N-terminally tagged with FLAG. Baculovirus-expressed proteins were used to co-infect SF9 insect cells. Insect cell pellets were dissolved in a buffer containing 25 mM Tris pH 8.0, 300 mM NaCl, 0.5 mM TCEP, complete EDTA-free protease inhibitors (Roche), and 0.1% NP-40. The supernatant of the cell lysate was incubated with M2 antibody resin (Sigma). The resin was washed on a chromatography column and eluted with 0.2 mg/ml FLAG peptide. The eluate was incubated overnight at 4°C in omni-leave nuclease (Epicentre Technologies), then concentrated and loaded onto a Superdex 200 column (GE Healthcare). The Superdex 200 column was eluted with 25 mM Tris pH 8.0, 150 mM NaCl, and 0.5 mM TCEP, and the fractions containing the PRC2 complex were pooled.

Nucleosome detection procedure : (The same procedure was used for WT and mutant EZH2Y6412N detection)

A. Compound Preparation

1. Prepare 10 mM stock solutions from solid material in 100% DMSO.

2. Serial 2- or 3-fold dilutions of 10 mM compound stocks in 100% DMSO were made to prepare compounds for an 11-point dose response.

B. Reagent Preparation

1. Prepare 1x assay buffer (containing 100 mM Tris pH 8.5, 4 mM DTT, and 0.01% Tween 20)

2. Purified HeLa oligonucleosomes and recombinant histone H1 (New England Biolabs) were diluted 1.67x in assay buffer.

3. Dilute the four protein complexes of PRC2 (EZH2, EED, SUZ12, RbAp48) to 3.5x in assay buffer.

4. Prepare a 10x ³ H SAM solution in assay buffer using 0.94 μCi/well of radioactive SAM (Perkin Elmer) and sufficient unlabeled SAM (Sigma) to a final concentration of 1.5 μM.

5. Dilute TCA to 20% with deionized water.

C. Enzyme reaction

1. Final reaction conditions were: PRC2 four-protein complex (4 nM when using WT EZH2, or 6 nM when using Y641N mutant EZH2), 1.5 μM SAM, 25 μg/mL oligonucleosomes, 50 nM rH1, and a reaction volume of 50 μl.

2. Add 1 μl of diluted compound to the test plate (96-well V-bottom polypropylene plate) or add 1 μl of DMSO to the control wells.

3. Add 30 μl of nucleosomes to the assay plate.

4. Add 14 μl of WT or Y641N mutant PRC2 four-protein complex to the assay plate.

5. Add 5 μl ³ H SAM to start the reaction.

6. After 60 minutes, add 100 μl of 20% TCA to terminate the reaction.

7. Transfer 150 μl of the stopped reaction mixture to the prepared filter plate (Millipore #MSIPN4B10).

8. Apply vacuum to the filter plate to filter the reaction mixture through the membrane.

9. Wash the filter plate with 5 x 200 μl PBS, blot dry, and dry in an oven for 30 minutes.

10. Add 50 μl of microscint-20 scintillation fluid (Perkin Elmer) to each well, wait for 30 minutes, and count using a liquid scintillation counter.

11. Some compounds were tested under high SAM conditions. In this case, the assay was performed as described above, but the reaction mixture contained 15 μM SAM. SAM was added to the assay plate at a 3.3x stock concentration for a total of 14.5 μCi/well.

D. Data Analysis

1. _IC50 values were determined by fitting the data to a four-parameter _IC50 equation using dedicated curve fitting software.

2. For compounds tested under high SAM conditions, the dose-response curves were fitted to a competitive inhibition model using dedicated curve fitting software to obtain K _i ^app values.

Preparation of HeLa oligonucleosomes:

Reagents

–Cell pellet: 15L HeLa S3 (Accelgen) + 6L HeLa S3 (homemade)

–Mnase (Worthington Biochemicals)

equipment

–SW-28 Rotor

–Dounce Homogenizer/B Pestle

buffer

– Lysis buffer: 20 mM Hepes pH 7.5, 0.25 M sucrose, 3 mM MgCl ₂ , 0.5% Nonidet P-40, 0.5 mM TCEP, 1 Roche protease tablet

-B: 20 mM Hepes pH 7.5, 3 mM MgCl ₂ , 0.5 mM EDTA, 0.5 mM TCEP, 1 Roche protease tablet

–MSB: 20 mM Hepes pH 7.5, 0.4 M NaCl, 1 mM EDTA, 0.5% v/v glycerol, 0.5 mM TCEP, 0.2 mM PMSF

–LSB: 20mM Hepes pH 7.5, 0.1M NaCl, 1mM EDTA, 0.5mM TCEP, 0.2mM PMSF

–NG: 20mM Hepes pH 7.5, 1mM EDTA, 0.4M NaCl, 0.2mM PMSF, 0.5mM TCEP

– Storage buffer: 20 mM Hepes pH 7.5, 1 mM EDTA, 10% glycerol, 0.2 mM PMSF, 0.5 mM TCEP

Operating Procedures

A.cell nucleus

1. Resuspend ~10 L of cell pellet in 2 x 40 mL of lysis buffer using a dounce homogenizer

2. Spin at 3000xg for 15 minutes

3. Repeat 2 times

4. Resuspend the cell pellet in 2 x 40 mL of B

5. Spin at 3000xg for 15 minutes

B. Resuspension of Nuclei

1. Resuspend the cell pellet in 2 x 40 mL MSB and spin at 5000 x g for 20 minutes

2. Resuspend the cell pellet in 2 x 15 mL HSB

3. Mix and homogenize 40 strokes to cut DNA

4. Spin the cell pellet at 10,000 x g for 20 minutes

5. Dialyze overnight (O/N) in LSB at 4°C. Only batch A was dialyzed in 50 nM NaCl in LSB for 3 hours.

C. Mnase digestion

Test MNase digestion (200 μl)

1. Raise the temperature to 37°C and maintain for 5 minutes

2. Add CaCl ₂ to 3 mM and add 10 U Mnase

3. Maintain at 37°C for 30 minutes, sampling 25 μL every 5 minutes

4. React with 1 μL 0.5M EDTA, 40 μL H ₂ O, 15 μL 10% SDS, 10 μL 5M NaCl and 100 μL phenol-chloroform, vortexing after each addition.

5. Spin at 13k for 5 minutes

6. Perform 5 μL aqueous phase reaction on 1% agarose gel

7. After a period of time, a ~2kb fragment was obtained

8. To scale up, select 15 minutes for A&B and 20 minutes for C&D

Add NaCl to 0.6M

D. Sucrose Gradient 1

1. Pour 6 x 34 mL of NG solution (using AKTA purifier) with a gradient of 5 to 35% sucrose into a 38.5 mL pollyallomer tube

2. Direct ~4.0 mL onto the top of the MN1 digest

3. Rotate at 26k for 16 hours at 4°C

4. Take 2 mL fraction from the top

5. Run on Page Gel

6. Dialyze fractions 7-14 0/N in 4 L LSB at 4°C, except for batch D, for 2 x 2 hours.

7. Repeat 3 times

E.Finally

1. Mix all in Amicon and concentrate (a little cloudy)

2. Add 10% glycerol

3. Spin at 5K for 15 minutes

4. 1.8 mg/mL contains 80 mL, totaling 144 mg

Biological activity

The biological activities of selected examples in the EZH2 nucleosome assay are provided in Table 3. Data are presented as _IC50 values (μM) or _Kjapp ^values (μM) for WT and mutant Y641N EZH2, as indicated.

Table 2

The entire contents of all publications and patent applications cited in this specification are incorporated herein by reference. Although the present invention has been described in considerable detail by way of illustration and example, it will be apparent to those skilled in the art that certain changes and modifications may be made in accordance with the teachings of the present invention without departing from the spirit or scope of the appended claims.

Claims (24)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, in: ^R1 is selected from the group consisting of: H, F, _C1 - _C4 alkyl, _C1 - _C4 alkoxy, C(O) ^R5 , _C3 - _C8 cycloalkyl, 3- to 12-membered heterocyclic and 5- to 12-membered heteroaryl, wherein each of the _C1 - _C4 alkyl or _C1 - _C4 alkoxy is optionally substituted by one or more ^R6 , and each of the _C3 - _C8 cycloalkyl, 3- to 12-membered heterocyclic or 5- to 12-membered heteroaryl is optionally substituted by one or more ^R7 ; ^R2 is H, F, or _C1 - _C4 alkyl; L is a bond or a _C1 - _C4 alkylene group; ^R3 is selected from the group consisting of: _C1 - _C4 alkyl, _C1 - _C4 alkoxy, OH, CN, C(O) ^R8 , ^COOR9 , ^NR10 , ^R11 , ^OR12 , _C3 -C8 cycloalkyl, 3- to ₁₂ -membered heterocyclic and 5- to 12-membered heteroaryl, wherein each of the _C1 - _C4 alkyl or _C1 - _C4 alkoxy is optionally substituted by one or more ^R6 , and each of the _C3 - _C8 cycloalkyl, 3- to 12-membered heterocyclic or 5- to 12-membered heteroaryl is optionally substituted by one or more ^R7 ; ^R4 is H, halogen, or _C1 - _C4 alkyl, wherein each of the _C1 - _C4 alkyl groups is optionally substituted by one or more ^R6 ; ^R5 is a _C1 - _C4 alkyl group, wherein each of the _C1 - _C4 alkyl groups is optionally substituted by one or more ^R14 groups; Each ^R6 is independently an OH, F, CN, or _C1 - _C4 alkoxy group; Each ^R7 is independently a _C1 - _C4 alkyl, OH, F, CN, _C1 - _C4 alkoxy, =O, CHO, C(O) ^R13 , _SO2R13 or ^a 3 to 6-membered heterocyclic group; ^R8 is a _C1 - _C4 alkyl group, wherein each of the _C1 - _C4 alkyl groups is optionally substituted by one or more ^R14 groups; ^R9 is H or _C1 - _C4 alkyl, wherein each of the _C1 - _C4 alkyl groups is optionally substituted by one or more ^R14 ; ^R10 and ^R11 are independently H or _C1 - _C4 alkyl, wherein each of the _C1 - _C4 alkyl groups is optionally substituted by one or more ^R14 ; R ¹² is selected from the group consisting of: C ₃ -C ₈ cycloalkyl, 3- to 12-membered heterocyclic and 5- to 12-membered heteroaryl, wherein each of the C ₃ -C ₈ cycloalkyl, 3- to 12-membered heterocyclic or 5- to 12-membered heteroaryl is optionally substituted by one or more R ⁷ ; Each R ¹³ is independently a _C1 - _C4 alkyl group, wherein each _C1 - _C4 alkyl group is optionally substituted by one or more R ¹⁵ groups; ^R14 and ^R15 are each independently OH, F, CN, or _C1 - _C4 alkoxy groups; and X and Z are independently _C1 - _C4 alkyl, _C1 - _C4 fluoroalkyl, _C1 - _C4 alkoxy, or _C1 - _C4 fluoroalkoxy. 2. The compound or salt of claim 1, wherein ^R2 is H. 3. The compound or salt of claim 1 or 2, wherein ^R4 is Cl, F, Br or _CH3 . 4. The compound or salt of claim 1, 2 or 3, wherein X is _CH3 , _OCH3 or _OCHF2 , and Z is _CH3 . 5. The compound or salt of any one of claims 1 to 4, wherein ^R1 is a _C1 - _C4 alkoxy group optionally substituted with one or more ^R6 . 6. The compound or salt of claim 5, wherein the _C1 - _C4 alkoxy group is _OCH3 . 7. The compound or salt of any one of claims 1 to 4, wherein ^R1 is a _C1 - _C4 alkyl group optionally substituted with one or more ^R6 . 8. The compound or salt of any one of claims 1 to 7, wherein L is a bond and ^R3 is a 3- to 12-membered heterocyclic group optionally substituted with one or more ^R7 . 9. The compound or salt of claim 8, wherein the 3 to 12-membered heterocyclic group is selected from the group consisting of: oxobutyryl, tetrahydrofuranyl, and tetrahydropyranyl, each optionally substituted with one or more R ⁷ . 10. A compound of formula (I-A) or a pharmaceutically acceptable salt thereof, in: ^R1 is a _C1 - _C4 alkoxy group; ^R2 is H; L is a bond; R ³ is a 3- to 12-membered heterocyclic group, which may optionally be replaced by one or more R ⁷ ; ^R4 is H or Cl; ^R7 is independently a _C1 - _C4 alkyl, OH, F, CN, _C1 - _C4 alkoxy, =O, CHO, C(O) ^R13 , _SO2R13 or ^a 3 to 6-membered heterocyclic group; Each of ^R13 is independently a _C1 - _C4 alkyl group, wherein each of the _C1 - _C4 alkyl groups is optionally substituted by one or more ^R15 groups; ^R15 are each independently OH, F, CN, or _C1 - _C4 alkoxy; and X and Z are independently _C1 - _C4 alkyl, _C1 - _C4 fluoroalkyl, _C1 - _C4 alkoxy, or _C1 - _C4 fluoroalkoxy. 11. The compound or salt of claim 10, wherein ^R3 is a 3- to 12-membered heterocyclic group selected from the group consisting of: oxobutyl, tetrahydrofuranyl, and tetrahydropyranyl, each optionally substituted by one or more ^R7 . 12. A compound that is 5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[(R)-methoxy(oxetane-3-yl)methyl]-3,4-dihydroisoquinoline-1(2H)-one, or a pharmaceutically acceptable salt thereof. 13. A compound that is 5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[(R)-methoxy(oxetane-3-yl)methyl]-3,4-dihydroisoquinoline-1(2H)-one. 14. A compound that is 5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[methoxy(oxetane-3-yl)methyl]-3,4-dihydroisoquinoline-1(2H)-one, or a pharmaceutically acceptable salt thereof. 15. A compound that is 5,8-dichloro-2-[(4-methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl]-7-[(S)-methoxy(oxetane-3-yl)methyl]-3,4-dihydroisoquinoline-1(2H)-one, or a pharmaceutically acceptable salt thereof. 16. A compound of formula (III) or a pharmaceutically acceptable salt thereof, in: ^R1 and ^R3 together form a 3 to 12-membered heterocyclic group that may be optionally replaced by one or more ^R7s ; ^R2 is H, F, or _C1 - _C4 alkyl; ^R4 is H, halogen, or _C1 - _C4 alkyl, wherein each of the _C1 - _C4 alkyl groups is optionally substituted by one or more ^R6 ; ^R6 is independently OH, F, CN or _C1 - _C4 alkoxy; ^R7 is independently a _C1 - _C4 alkyl, OH, F, CN, _C1 - _C4 alkoxy, =O, CHO, C(O) ^R13 , _SO2R13 or ^a 3 to 6-membered heterocyclic group; Each of ^R13 is independently a _C1 - _C4 alkyl group, wherein each of the _C1 - _C4 alkyl groups is optionally substituted by one or more ^R15 groups; ^R15 are each independently OH, F, CN, or _C1 - _C4 alkoxy; and X and Z are independently _C1 - _C4 alkyl, _C1 - _C4 fluoroalkyl, _C1 - _C4 alkoxy, or _C1 - _C4 fluoroalkoxy. 17. The compound or salt of claim 16, wherein ^R1 and ^R3 together form a 3- to 12-membered heterocyclic group selected from the group consisting of: aziridine, pyrrolidinyl, piperidinyl, and homopiperidinyl, each optionally substituted by one or more ^R7 . 18. The compound or salt of claim 17, wherein ^R7 is CHO, C(O) ^R13 or _SO2R13 , and each ^R13 is independently ^a _C1 - _C4 alkyl group optionally substituted with one or more ^R15 . 19. A pharmaceutical composition comprising a compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. 20. Use of a compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating abnormal cell growth in a patient. 21. The use of claim 20, wherein the abnormal cell growth is cancer. 22. The use of claim 20 or 21, wherein the patient is a person. 23. The compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, for the treatment of abnormal cell growth in a patient. 24. A combination of a compound of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof with one or more other anticancer agents.