HK1241705B - Therapeutic inhibitory compounds - Google Patents

Description

Therapeutic inhibitory compounds

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. application serial number 62/025,203, filed on July 16, 2014, U.S. application serial number 62/187,786, filed on July 1, 2015, U.S. application serial number 62/190,223, filed on July 8, 2015, and international application PCT/US2014/072851, filed on December 30, 2014, each of which is incorporated herein by reference in its entirety.

Background Art

There is a need in the medical field for effective treatments for diseases and conditions associated with the vascular system. Such diseases and conditions include, but are not limited to, angioedema, macular edema, and cerebral edema.

Summary of the Invention

Provided herein are heterocyclic derivative compounds and pharmaceutical compositions comprising the same.The subject compounds and compositions are useful for inhibiting plasma kallikrein.

One embodiment provides a compound, or a pharmaceutically acceptable salt thereof, having the structure of Formula (I):

in,

Ring A is an optionally substituted bicyclic heteroaryl ring;

Ring B is an optionally substituted monocyclic heteroaryl ring or an optionally substituted bicyclic heteroaryl ring;

each R ¹² , R ¹³ or R ¹⁴ is independently selected from hydrogen, cyano, halo, hydroxy, azido, amino, nitro, —CO ₂ H, —S(O)—R ²⁰ , —SR ²⁰ , —S(O) ₂ —R ²⁰ , optionally substituted alkoxy, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted (heterocyclyl)-O—, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted alkylamino, optionally substituted dialkylamino, —CO—R ²⁰ , —CO ₂ —R ²⁰ , —CO(NR ²¹ ) ₂ , —SO ₂ (NR ²¹ ) ₂ , —C(═NR ²² )—(NR ²¹ ) ₂ , or optionally substituted alkynyl;

each R ¹ or R ² is independently selected from hydrogen, halo, hydroxy, amino, -CO ₂ H, -S(O)-R ²⁰ , -SR ²⁰ , -S(O) ₂ -R ^{20 , optionally substituted alkoxy, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted (heterocyclyl)-O-, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted alkylamino, optionally substituted dialkylamino, -CO-R 20 , -CO 2 -R 20} , -CO(NR 21 ) 2 , -SO 2 (NR 21 ) 2 , -C(═NR 22 )-(NR 21 ) 2 or optionally substituted alkynyl; or optionally, R 1 and R 2 are independently selected from hydrogen, halo, hydroxy, amino, -CO 2 H, -S(O)-R ^{20 , -SR 20 , -S(O) 2 -R 20 , optionally substituted alkoxy, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted (heterocyclyl)-O-, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted alkylamino, optionally substituted dialkylamino, -CO-R 20} , -CO ₂ -R ²⁰ , -CO(NR ²¹ ) ₂ , -SO ₂ (NR ²¹ ) ₂ , -C(═NR ²² )-(NR ²¹ ) ₂ or optionally substituted alkynyl ^{; R 2} is an optionally substituted C1-C5 alkyl group and is linked together to form a ring; or optionally, R ¹ and R ² together form an oxo group;

each R ³ or R ⁴ is independently selected from hydrogen, -CO ₂ H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, -CO-R ²⁰ , -CO ₂ -R ²⁰ , -CO(NR ²¹ ) ₂ , -SO ₂ (NR ²¹ ) ₂ , -C(＝NR ²² )-(NR ²¹ ) ₂ or optionally substituted alkynyl; or optionally, R ³ and R ⁴ are optionally substituted C1-C5 alkyl and are linked to form a ring;

each R ²⁰ is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl;

each R ²¹ is selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl; and

each R ²² is selected from hydrogen, -CN, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclyl; with the proviso that the compound of formula (I) is not 2-[[4,5,6,7-tetrahydro-3-(trifluoromethyl)-1H-indazol-1-yl]methyl]-N-(2-thienylmethyl)-4-pyridinecarboxamide.

One embodiment provides a compound, or a pharmaceutically acceptable salt thereof, having the structure of Formula (Ia):

in,

Ring A is an optionally substituted bicyclic heteroaryl ring; and

Ring B is an optionally substituted monocyclic heteroaryl ring or an optionally substituted bicyclic heteroaryl ring;

with the proviso that the compound of formula (Ia) is not 2-[[4,5,6,7-tetrahydro-3-(trifluoromethyl)-1H-indazol-1-yl]methyl]-N-(2-thienylmethyl)-4-pyridinecarboxamide.

One embodiment provides a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

One embodiment provides a method of inhibiting kallikrein comprising contacting kallikrein with a compound of formula (I).

One embodiment provides a method of treating angioedema in a patient in need thereof, comprising administering to the patient a composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

One embodiment provides a pharmaceutical composition comprising a compound of formula (Ia), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

One embodiment provides a method of inhibiting kallikrein comprising contacting kallikrein with a compound of formula (Ia).

One embodiment provides a method of treating angioedema in a patient in need thereof, comprising administering to the patient a composition comprising a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof.

Incorporation by reference

All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference for the specific purposes described herein.

DETAILED DESCRIPTION

As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly indicates otherwise. Thus, for example, reference to "an agent" includes a plurality of such agents, and reference to "the cell" includes reference to one or more cells (or cells) and equivalents thereof known to those skilled in the art, and so on. When a range is used herein for a physical property, such as a molecular weight, or a chemical property, such as a chemical formula, all combinations and subcombinations of that range and specific embodiments thereof are intended to be included. When the term "about" refers to a number or a numerical range, it means that the number or numerical range referred to is an approximate value within the experimental variability (or within the statistical experimental error), so that in some cases, the number or numerical range will vary between 1% and 15% of the specified number or numerical range. The term "comprising" (and related terms, such as "including" or "having") is not intended to exclude certain other embodiments, for example, embodiments of any combination of the substances, compositions, methods, or processes described herein, "consisting of" or "consisting essentially of" the features.

definition

As used in the specification and the appended claims, unless otherwise specified, the following terms have the meanings indicated below.

"Amino" refers to a _-NH2 group.

"Cyano" refers to a -CN group.

"Nitro" refers to the _-NO2 radical.

"Oxa" refers to an -O- group.

"Oxo" refers to the =0 radical.

"Thio" refers to a =S group.

"Imino" refers to the =N-H group.

"Oximo" refers to a =N-OH group.

"Hydrazino" refers to the =N- _NH2 radical.

"Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, and having 1-15 carbon atoms (e.g., _C1 - _C15 alkyl). In certain embodiments, the alkyl group contains 1-13 carbon atoms (e.g., _C1 - _C13 alkyl). In certain embodiments, the alkyl group contains 1-8 carbon atoms (e.g., _C1 - _C8 alkyl). In other embodiments, the alkyl group contains 1-5 carbon atoms (e.g., _C1 - _C5 alkyl). In other embodiments, the alkyl group contains 1-4 carbon atoms (e.g., _C1 - _C4 alkyl). In other embodiments, the alkyl group contains 1-3 carbon atoms (e.g., _C1 - _C3 alkyl). In other embodiments, the alkyl group contains 1-2 carbon atoms (e.g., _C1 - _C2 alkyl). In other embodiments, the alkyl group contains 1 carbon atom (e.g., _C1 alkyl). In other embodiments, the alkyl group contains 5-15 carbon atoms (e.g., _C5 - _C15 alkyl). In other embodiments, the alkyl group contains 5-8 carbon atoms (e.g., _C5 - _C8 alkyl). In other embodiments, the alkyl group contains 2-5 carbon atoms (e.g., _C2 - _C5 alkyl). In other embodiments, the alkyl group contains 3-5 carbon atoms (e.g., _C3 - _C5 alkyl). In other embodiments, the alkyl group is selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (isopropyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (isobutyl), 1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl). The alkyl group is attached to the rest of the molecule by a single bond. Unless stated otherwise specifically in the specification, an alkyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximino, trimethylsilanyl, ^-ORa , ^-SRa , -OC(O) ^-Ra , -N( ^Ra ) ₂ , -C(O) ^Ra , -C(O) ^ORa , -C(O)N( ^Ra ) ₂ , -N( ^Ra )C(O) ^ORa , -OC(O) ^-N ( ^Ra ) ₂ , -N( ^Ra )C(O) ^Ra , -N( ^Ra )S(O) _tRa (where t is 1 or 2), -S( ^O)tORa ₍ where t is 1 or 2), -S(O) _tRa ⁽ where t is 1 or 2), and -S(O) _tN ( ^Ra ) ₂ (where t is 1 or 2), where each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl).

"Alkoxy" refers to a group of the general formula -O-alkyl bonded through an oxygen atom, wherein alkyl is an alkyl chain as defined above.

"Alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and having 2-12 carbon atoms. In certain embodiments, the alkenyl group contains 2-8 carbon atoms. In other embodiments, the alkenyl group contains 2-4 carbon atoms. The alkenyl group is connected to the rest of the molecule by a single bond, for example, vinyl, prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, pent-1,4-dienyl, etc. Unless stated otherwise specifically in the specification, an alkenyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximino, trimethylsilanyl, ^-ORa , -SRa, -OC(O) ^-Ra , ^-N ( ^Ra ), -C(O) _Ra , -C(O ^{) ORa} , -C(O)N( ^Ra ), -N( ^Ra )C(O) ^ORa , -OC(O)-N( ^Ra _{) ,} ^-N ( ^Ra )C(O) ^Ra , -N( ^Ra )S(O) _tRa (where t is 1 or 2), ^-S(O)tORa ₍ where t is 1 or 2), -S(O) _tRa (where ^t is 1 or 2), and -S(O) _tN ( ^Ra ) ₂ (where t is 1 or 2), where each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl).

"Alkynyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, and having 2-12 carbon atoms. In certain embodiments, an alkynyl group contains 2-8 carbon atoms. In other embodiments, an alkynyl group contains 2-6 carbon atoms. In other embodiments, an alkynyl group contains 2-4 carbon atoms. An alkynyl group is attached to the rest of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless stated otherwise specifically in the specification, an alkynyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximino, trimethylsilanyl, ^-ORa , -SRa, -OC(O) ^-Ra , ^-N ( ^Ra ) ₂ , -C(O) ^Ra , -C(O) ^ORa , -C(O)N( ^Ra ) ₂ , -N( ^Ra )C(O) ^ORa , -OC(O)-N( ^Ra ) ₂ ^, -N( ^Ra )C(O) ^Ra , -N( ^Ra )S(O) _tRa (where t is 1 or 2), -S(O ^)tORa ₍ where t is 1 or 2), -S(O) _tRa (where ^t is 1 or 2), and -S(O) _tN ( ^Ra ) ₂ (where t is 1 or 2), where each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl).

"Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain that connects the rest of the molecule to a radical, consisting solely of carbon and hydrogen, containing no unsaturation, and having 1-12 carbon atoms, for example, methylene, ethylene, propylene, n-butylene, etc. The alkylene chain is connected to the rest of the molecule and to the radical by a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical are through one carbon in the alkylene chain or through any two carbons in the chain. In certain embodiments, the alkylene group contains 1-8 carbon atoms (e.g., _C1 - _C8 alkylene). In other embodiments, the alkylene group contains 1-5 carbon atoms (e.g., _C1 - _C5 alkylene). In other embodiments, the alkylene group contains 1-4 carbon atoms (e.g., _C1 - _C4 alkylene). In other embodiments, the alkylene group contains 1-3 carbon atoms (e.g., _C1 - _C3 alkylene). In other embodiments, the alkylene group comprises 1-2 carbon atoms (e.g., C ₁ -C ₂ alkylene). In other embodiments, the alkylene group comprises 1 carbon atom (e.g., C ₁ alkylene). In other embodiments, the alkylene group comprises 5-8 carbon atoms (e.g., C ₅ -C ₈ alkylene). In other embodiments, the alkylene group comprises 2-5 carbon atoms (e.g., C ₂ -C ₅ alkylene). In other embodiments, the alkylene group comprises 3-5 carbon atoms (e.g., C ₃ -C ₅ alkylene). Unless stated otherwise specifically in the specification, an alkylene chain is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximino, trimethylsilanyl, ^-ORa , -SRa, -OC(O) ^-Ra , ^-N ( ^Ra ) ₂ , -C(O) ^Ra , -C(O) ^ORa , -C(O)N( ^Ra ) ₂ , -N( ^Ra )C(O) ^ORa , -OC(O)-N( ^Ra ) ₂ ^, -N( ^Ra )C(O) ^Ra , -N( ^Ra )S(O) _tRa (where t is 1 or 2), -S(O ^)tORa ₍ where t is 1 or 2), -S(O) _tRa (where t is 1 or 2), and ^-S (O) _tN ( ^Ra ) ₂ (where t is 1 or 2), where each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl).

"Alkynylene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain that connects the rest of the molecule to a radical, consisting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond, and having 2-12 carbon atoms. The alkynylene chain is connected to the rest of the molecule by a single bond and to the radical by a single bond. In certain embodiments, the alkynylene group comprises 2-8 carbon atoms (e.g., _C2 - _C8 alkynylene). In other embodiments, the alkynylene group comprises 2-5 carbon atoms (e.g., _C2 - _C5 alkynylene). In other embodiments, the alkynylene group comprises 2-4 carbon atoms (e.g., _C2 - _C4 alkynylene). In other embodiments, the alkynylene group comprises 2-3 carbon atoms (e.g., _C2 - _C3 alkynylene). In other embodiments, the alkynylene group comprises 2 carbon atoms (e.g., _C2 alkynylene). In other embodiments, the alkynylene group comprises 5-8 carbon atoms (e.g., _C5 - _C8 alkynylene). In other embodiments, the alkynylene group comprises 3-5 carbon atoms (eg, C ₃ -C ₅ alkynylene). Unless stated otherwise specifically in the specification, an alkynylene chain is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximino, trimethylsilanyl, ^-ORa , -SRa ^, -OC(O) ^-Ra , -N( ^Ra ) ₂ , -C(O)Ra, -C(O) ^ORa , ^-C (O)N( ^Ra ) ₂ , -N( ^Ra )C(O) ^ORa , -OC(O)-N( ^Ra ) ₂ ^, -N( ^Ra )C(O) ^Ra , -N( ^Ra )S(O) _tRa (where t is 1 or 2), -S(O) _tORa (where t is 1 or 2), -S(O) _tRa (where t is 1 or 2) ^, and ^-S (O) _tN ( ^Ra ) ₂ (where t is 1 or 2), where each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl).

"Aryl" refers to a radical derived from an aromatic monocyclic or polycyclic hydrocarbon ring system by removing hydrogen atoms from ring carbon atoms. An aromatic monocyclic or polycyclic hydrocarbon ring system contains only hydrogen and 5-8 carbon atoms, wherein at least one ring of the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2)π electron system according to Hückel theory. Ring systems from which aryl radicals are derived include, but are not limited to, radicals such as benzene, fluorene, indane, indene, tetralin, and naphthalene. Unless stated otherwise specifically in the specification, the term “aryl” or the prefix “ar” (as in “aralkyl”) is meant to include aryl groups optionally substituted by one or more substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, halo, fluoroalkyl, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -Rb ^{- ORa} , -Rb-OC(O)-Ra, -Rb ^{- OC} (O) ^-ORa , -Rb- ^OC (O)-N( ^Ra ) ₂ , -Rb-N( ^Ra ) ₂ , -Rb ^{- C} (O) ^Ra , -Rb- ^C (O) ^ORa , -Rb-C(O) ^{N ( Ra} ) ₂ , -R ^b -OR ^c -C(O)N(R ^a ) ₂ , -R ^b -N(R ^a )C(O)OR ^a , -R ^b -N(R ^a )C(O)R ^a , -R ^b -N(R ^a )S(O) _t R ^a (wherein t is 1 or 2), -R ^b -S(O) _t R ^a (wherein t is 1 or 2), -R ^b -S(O) _t OR ^a (wherein t is 1 or 2), and -R ^b -S(O) _t N(R ^a ) ₂ (wherein t is 1 or 2), wherein each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), each R ^b is independently a direct bond or a straight or branched alkylene or alkenylene chain, and R ^c is a straight or branched alkylene or alkenylene chain, and unless otherwise specified, each of the above substituents is unsubstituted.

"Aralkyl" refers to a group of the formula ^-Rc -aryl, where ^Rc is an alkylene chain as defined above, for example, methylene, ethylene, and the like. The alkylene chain portion of the aralkyl group is optionally substituted as described above for an alkylene chain. The aryl portion of the aralkyl group is optionally substituted as described above for an aryl group.

"Aralkenyl" refers to a group of the formula ^-Rd -aryl, where ^Rd is an alkenylene chain as defined above. The aryl portion of the aralkenyl group is optionally substituted as described above for aryl groups. The alkenylene chain portion of the aralkenyl group is optionally substituted as described above for alkenylene groups.

"Aralkynyl" refers to a group of the formula ^-Re -aryl, where ^Re is an alkynylene chain as defined above. The aryl portion of the aralkynyl group is optionally substituted as described above for aryl groups. The alkynylene chain portion of the aralkynyl group is optionally substituted as described above for alkynylene chains.

"Aralkyloxy" refers to a radical of the formula -ORc- ^aryl , bonded through an oxygen atom, wherein ^Rc is an alkylene chain as defined above, for example, methylene, ethylene, and the like. The alkylene chain portion of the aralkyl radical is optionally substituted as described above for alkylene chains. The aryl portion of the aralkyl radical is optionally substituted as described above for aryl groups.

"Carbocyclyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting only of carbon atoms and hydrogen atoms, including fused or bridged ring systems having 3-15 carbon atoms. In certain embodiments, the carbocyclyl comprises 3-10 carbon atoms. In other embodiments, the carbocyclyl comprises 5-7 carbon atoms. The carbocyclyl is connected to the rest of the molecule via a single bond. The carbocyclyl is saturated (i.e., containing only a single C-C bond) or unsaturated (i.e., containing one or more double or triple bonds). Fully saturated carbocyclyls are also referred to as "cycloalkyls." Examples of monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Unsaturated carbocyclyls are also referred to as "cycloalkenyls." Examples of monocyclic cycloalkenyls include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycyclic carbocyclyl groups include, for example, adamantyl, norbornyl (ie, bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless stated otherwise specifically in the specification, the term "carbocyclyl" is intended to include carbocyclyl groups optionally substituted by one or more substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -Rb ^{- ORa} , -Rb-OC(O)-Ra, -Rb ^-OC (O) ^-ORa , -Rb- ^{OC (} O)-N( ^Ra ) ₂ , -Rb ^- N(Ra) ₂ , -Rb- ^C ( ^O ) ^Ra , ^-Rb - ^C (O) ^ORa , -Rb- ^C (O)N( ^Ra ) ₂ , ^-Rb- -ORc- ^C (O)N(R ^a ) ₂ , -Rb ^- N(R ^a )C(O)OR ^a , -Rb- ^N (R ^a )C(O)R ^a , -Rb- ^N (R ^a )S(O) _t R ^a (wherein t is 1 or 2), -Rb- ^S (O) _t R ^a (wherein t is 1 or 2), -Rb- ^S (O) _t OR ^a (wherein t is 1 or 2), and -Rb- ^S (O) _t N(R ^a ) ₂ (wherein t is 1 or 2), wherein each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), each R ^b is independently a direct bond or a straight or branched alkylene or alkenylene chain, and R ^c is a straight or branched alkylene or alkenylene chain, and unless otherwise specified, each of the above substituents is unsubstituted.

"Carbocyclylalkyl" refers to a radical of the formula ^-Rc -carbocyclyl, wherein ^Rc is an alkylene chain as defined above. The alkylene chain and the carbocyclyl radical are optionally substituted as described above.

"Carbocyclylalkynyl" refers to a radical of the formula ^-Rc -carbocyclyl, wherein ^Rc is an alkynylene chain as defined above. The alkynylene chain and the carbocyclyl radical are optionally substituted as described above.

"Carbocyclylalkoxy" refers to a radical of the formula ^-ORc -carbocyclyl, bonded through an oxygen atom, wherein ^Rc is an alkylene chain as defined above. The alkylene chain and the carbocyclyl radical are optionally substituted as described above.

As used herein, "carboxylic acid bioisostere" refers to a functional group or moiety that exhibits physical, biological, and/or chemical properties similar to a carboxylic acid moiety. Examples of carboxylic acid bioisosteres include, but are not limited to

wait.

"Halo" or "halogen" refers to bromo, chloro, fluoro, or iodo substituents.

"Fluoroalkyl" refers to an alkyl group as defined above substituted with one or more fluorinated groups as defined above, for example, trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. In some embodiments, the alkyl portion of the fluoroalkyl group is optionally substituted as described above for an alkyl group.

"Heterocyclyl" refers to a stable 3-18 membered non-aromatic ring radical containing 2-12 carbon atoms and 1-6 heteroatoms selected from nitrogen, oxygen and sulfur. Unless otherwise expressly provided in the specification, a heterocyclyl is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, optionally including fused or bridged ring systems. The heteroatoms in the heterocyclyl are optionally oxidized. If one or more nitrogen atoms are present, they are optionally quaternized. The heterocyclyl is partially or fully saturated. The heterocyclyl is connected to the rest of the molecule through any atom in the ring. Examples of such heterocyclic groups include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidineyl, thiazolidinyl, tetrahydrofuranyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. ,octahydroindolyl,octahydroisoindolyl,2-oxopiperazinyl,2-oxopiperidinyl,2-oxopyrrolidinyl,oxazolidinyl,piperidinyl,piperazinyl,4-piperidonyl,pyrrolidinyl,pyrazoli dinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thiomorpholinyl). Unless stated otherwise specifically in the specification, the term "heterocyclyl" is intended to include heterocyclyl groups as defined above optionally substituted by one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -Rb ^- ORa, -Rb-OC(O)-Ra, -Rb-OC(O) ^-ORa , -Rb ^{- OC} (O ⁾ -N(Ra) ₂ , -Rb ^{- N} ( ^Ra ) ₂ , -Rb ^-C ( ^O ) ^Ra , ^-Rb - ^C (O) ^ORa , -Rb- ^C (O)N( ^Ra ) ₂ , -R ^b -OR ^c -C(O)N(R ^a ) ₂ , -R ^b -N(R ^a )C(O)OR ^a , -R ^b -N(R ^a )C(O)R ^a , -R ^b -N(R ^a )S(O) _t R ^a (wherein t is 1 or 2), -R ^b -S(O) _t R ^a (wherein t is 1 or 2), -R ^b -S(O) _t OR ^a (wherein t is 1 or 2), and -R ^b -S(O) _t N(R ^a ) ₂ (wherein t is 1 or 2), wherein each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), each R ^b is independently a direct bond or a straight or branched alkylene or alkenylene chain, and R ^c is a straight or branched alkylene or alkenylene chain, and unless otherwise specified, each of the above substituents is unsubstituted.

"N-heterocyclyl" or "N-attached heterocyclyl" refers to a heterocyclyl as defined above that contains at least one nitrogen atom, and wherein the point of attachment of the heterocyclyl to the rest of the molecule is through the nitrogen atom in the heterocyclyl. N-heterocyclyl groups are optionally substituted as described above for heterocyclyl groups. Examples of such N-heterocyclyl groups include, but are not limited to, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

"C-heterocyclyl" or "C-attached heterocyclyl" refers to a heterocyclyl as defined above containing at least one heteroatom, and wherein the point of attachment of the heterocyclyl to the rest of the molecule is through a carbon atom in the heterocyclyl. C-heterocyclyl is optionally substituted as described above for heterocyclyl. Examples of such C-heterocyclyl groups include, but are not limited to, 2-morpholinyl, 2-, 3-, or 4-piperidinyl, 2-piperazinyl, 2- or 3-pyrrolidinyl, and the like.

"Heterocyclylalkyl" refers to a radical of the formula ^-Rc -heterocyclyl, where ^Rc is an alkylene chain as defined above. If the heterocyclyl radical is a nitrogen-containing heterocyclyl radical, the heterocyclyl radical is optionally attached to an alkyl radical at the nitrogen atom. The alkylene chain of the heterocyclylalkyl radical is optionally substituted as described above for an alkylene chain. The heterocyclyl portion of the heterocyclylalkyl radical is optionally substituted as described above for a heterocyclyl radical.

"Heterocyclylalkoxy" refers to a radical of the general formula ^-OR -heterocyclyl, bonded through an oxygen atom, wherein ^R is an alkylene chain as defined above. If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is optionally attached to an alkyl group at the nitrogen atom. The alkylene chain of the heterocyclylalkoxy radical is optionally substituted as described above for alkylene chains. The heterocyclyl portion of the heterocyclylalkoxy radical is optionally substituted as described above for heterocyclyl groups.

"Heteroaryl" refers to a group derived from a 3-18 membered aromatic ring radical containing 2-17 carbon atoms and 1-6 heteroatoms selected from nitrogen, oxygen and sulfur. As used herein, a heteroaryl group is a monocyclic, bicyclic, tricyclic or tetracyclic ring system in which at least one ring of the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2)π electron system according to the Huckel theory. Heteroaryl groups include fused or bridged ring systems. The heteroatoms in the heteroaryl group are optionally oxidized. If one or more nitrogen atoms are present, they are optionally quaternized. The heteroaryl group is attached to the rest of the molecule via any atom in the ring. Examples of heteroaryl groups include, but are not limited to, azalinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolanyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolanyl, benzodioxinyl, benzopyranyl, benzopyrone, benzofuranyl, benzofuranonyl, benzofuran ... , benzothiophenyl, benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl pyridyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyridazinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, dihydroindolinyl, isoindolinyl, isoquinolinyl, indolizinyl, isoxazolyl, 5,8- Methoxy-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyridine pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazole yl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl and thienyl (zepinyl, acridinyl, benzmidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo[d]thiazol yl,benzothiadiazolyl,benzo[b][1,4]dioxepinyl,benzo[b][1,4]oxazinyl,1,4-benzodioxanyl,benzonaphthofuranyl,benzoxazolyl,benzodioxolyl,benzodioxinyl,benzopyranyl,benzopy ranonyl,benzofuranyl,benzofuranonyl,benzothienyl(benzothiophenyl),benzothieno[3,2-d]pyrimidinyl,benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl,carbazolyl,cinnolinyl, cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl,5,6-dihydrobenzo[h]cinnolinyl,6,7-dihydro-5H-benzo[6,7 ]cyclohepta[1,2-c]pyridazinyl,dibenzofuranyl,dibenzothiophenyl,furanyl,furanonyl,furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5,6,7,8,9,10-hexah ydrocycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl,isothiazolyl,imidazolyl,indazolyl,indolyl,indazolyl,isoindolyl,indolinyl,isoindolinyl,isoquino lyl,indolizinyl,isoxazolyl,5,8-methano-5,6,7,8-tetrahydroquinazolinyl,naphthyridinyl,1,6-naphthyridinonyl,oxadiazolyl,2-oxoazepinyl,oxazolyl,oxiranyl,5,6,6a,7,8,9,10, 10a-octahydrobenzo[h]quinazolinyl,1-phenyl-1H-pyrrolyl,phenazinyl,phenothiazinyl,phenoxazinyl,phthalazinyl,pteridinyl,purinyl,pyrrolyl,pyrazolyl,pyrazolo[3,4-d]pyrimi dinyl,pyridinyl,pyrido[3,2-d]pyrimidinyl,pyrido[3,4-d]pyrimidinyl,pyrazinyl,pyrimidinyl,pyridazinyl,pyrrolyl,quinazolinyl,quinoxalinyl,quinolinyl,isoquinolinyl,tetrah ydroquinolinyl,5,6,7,8-tetrahydroquinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7 ,8-tetrahydropyrido[4,5-c]pyridazinyl,thiazolyl,thiadiazolyl,triazolyl,tetrazolyl,triazinyl,thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimidinyl,thieno[2,3-c]pridinyl,and thiophenyl(iethienyl)). Unless stated otherwise specifically in the specification, the term "heteroaryl" is intended to include heteroaryl groups as defined above optionally substituted by one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl, haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -Rb ^- ORa, -Rb-OC(O)-Ra, -Rb-OC(O ^{) -ORa} , -Rb ^{- OC} (O) ^-N (Ra) ₂ , -Rb- ^N ( ^Ra ) ₂ , -Rb ^{- C} ( ^O ) ^Ra , -Rb- ^C (O) ^ORa , ^-Rb -C(O)N(R ^a ) ₂ , -R ^b -OR ^c -C(O)N(R ^a ) ₂ , -R ^b -N(R ^a )C(O)OR ^a , -R ^b -N(R ^a )C(O)R ^a , -R ^b -N(R ^a )S(O) _t R ^a (wherein t is 1 or 2), -R ^b -S(O) _t R ^a (wherein t is 1 or 2), -R ^b -S(O) _t OR ^a (wherein t is 1 or 2), and -R ^b -S(O) _t N(R ^a ) ₂ (wherein t is 1 or 2), wherein each R ^a is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), each R ^b is independently a direct bond or a straight or branched alkylene or alkenylene chain, and R ^c is a straight or branched alkylene or alkenylene chain, and unless otherwise specified, each of the above substituents is unsubstituted.

"N-heteroaryl" refers to a heteroaryl group as defined above that contains at least one nitrogen atom and wherein the point of attachment of the heteroaryl group to the rest of the molecule is through the nitrogen atom in the heteroaryl group. N-heteroaryl groups are optionally substituted as described above for heteroaryl groups.

"C-heteroaryl" refers to a heteroaryl group as defined above wherein the point of attachment of the heteroaryl group to the rest of the molecule is through a carbon atom in the heteroaryl group. C-heteroaryl groups are optionally substituted as described above for heteroaryl groups.

"Heteroarylalkyl" refers to a radical of the formula ^-Rc -heteroaryl, where ^Rc is an alkylene chain as defined above. If the heteroaryl group is a nitrogen-containing heteroaryl group, the heteroaryl group is optionally attached to the alkyl group at the nitrogen atom. The alkylene chain of the heteroarylalkyl group is optionally substituted as described above for the alkylene chain. The heteroaryl portion of the heteroarylalkyl group is optionally substituted as described above for the heteroaryl group.

"Heteroarylalkoxy" refers to a radical of the formula -OR- ^heteroaryl , bonded through an oxygen atom, wherein ^R is an alkylene chain as defined above. If the heteroaryl group is a nitrogen-containing heteroaryl group, the heteroaryl group is optionally attached to an alkyl group at the nitrogen atom. The alkylene chain of the heteroarylalkoxy group is optionally substituted as described above for an alkylene chain. The heteroaryl portion of the heteroarylalkoxy group is optionally substituted as described above for a heteroaryl group.

In some embodiments, compounds disclosed herein contain one or more asymmetric centers, and thus produce enantiomers, diastereomers, and other stereoisomeric forms defined as (R)- or (S)- according to absolute stereochemistry. Unless otherwise stated, this disclosure is intended to encompass all stereoisomeric forms of compounds disclosed herein. When compounds as described herein contain olefin double bonds, unless otherwise stated, this disclosure is intended to include E and Z geometric isomers (e.g., cis or trans) simultaneously. Similarly, all possible isomers are also intended to be included, as well as their racemic forms and optically pure forms, and all tautomeric forms. The term "geometric isomer" refers to E or Z geometric isomers (e.g., cis or trans) of olefin double bonds. The term "positional isomer" refers to structural isomers around the central ring, such as ortho, meta, and para isomers around the phenyl ring.

"Tautomers" refers to molecules in which a proton can move from one atom of a molecule to another atom of the same molecule. In certain embodiments, the compounds described herein exist as tautomers. Where tautomerization is possible, a chemical equilibrium of the tautomers will exist. The exact ratio of the tautomers depends on several factors, including physical state, temperature, solvent, and pH. Some examples of tautomeric equilibrium include:

In some embodiments, the compounds disclosed herein are used in different isotopically enriched forms, for example, forms enriched in ² H, ³ H, ¹¹ C, ¹³ C, and/or ¹⁴ C content. In a specific embodiment, the compound is deuterated at at least one position. Such deuterated forms can be prepared by the procedures described in U.S. Patent Nos. 5,846,514 and 6,334,997. As described in U.S. Patent Nos. 5,846,514 and 6,334,997, deuteration can improve metabolic stability and/or efficacy, thereby increasing the duration of drug action.

Unless otherwise stated, structures depicted herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structure except for the replacement of a hydrogen by deuterium or tritium, or the replacement of a carbon by ¹³ C- or ¹⁴ C-enriched carbon are within the scope of this disclosure.

The compounds of the present disclosure optionally contain unnatural ratios of atomic isotopes at one or more atoms forming such compounds. For example, isotopes such as deuterium ( ² H), tritium ( ³ H), iodine-125 ( ¹²⁵ I) or carbon-14 ( ¹⁴ C) can be used to label the compounds. Isotopic substitutions with ² H, ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ C, ¹² N, ¹³ N, ¹⁵ N, ¹⁶ N, ¹⁶ O, ¹⁷ O, ¹⁴ F, 15 F, ¹⁶ F, ¹⁷ F, ¹⁸ F, ³³ S, ³⁴ S, ³⁵ S, ³⁶ S, ³⁵ Cl, ³⁷ Cl, ⁷⁹ Br, ⁸¹ Br, ¹²⁵ I are within the contemplation of the present invention. All isotopic variants of the compounds ^of the present invention, whether or not they are radioactive, are encompassed within the scope of the present invention.

In certain embodiments, some or all of the ¹ H atoms of the compounds disclosed herein are replaced with ² H atoms. Methods for synthesizing deuterium-containing compounds are known in the art and include, by way of non-limiting example only, the following synthetic methods.

Deuterium-substituted compounds are synthesized using a variety of methods, such as those described in Dean, Dennis C.; Editor. Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [In: Curr., Pharm. Des., 2000; 6(10)] 2000, 110pp; George W.; Varma, Rajender S. The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; and Evans, E. Anthony. Synthesis of radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32.

Deuterated starting materials are readily available and are amenable to the synthetic methods described herein to provide the synthesis of deuterium-containing compounds.Many deuterium-containing reagents and building blocks are commercially available from chemical suppliers such as Aldrich Chemical Co.

Deuterium transfer reagents suitable for nucleophilic substitution reactions, such as iodomethane-d ₃ (CD ₃ I), are readily available and can be used to transfer a deuterated carbon atom to a reaction substrate under nucleophilic substitution reaction conditions. By way of example only, the use of CD ₃ I is shown in the following reaction scheme.

Deuterium is transferred to the reaction substrate using a deuterium transfer reagent such as lithium aluminum deuteride (LiAlD ₄ ) under reducing conditions. By way of example only, the use of LiAlD ₄ is shown in the reaction scheme below.

By way of example only, as shown in the following reaction scheme, deuterium gas and a palladium catalyst are used to reduce unsaturated carbon-carbon bonds and perform reductive substitution of aromatic carbon-halogen bonds.

In one embodiment, the compounds disclosed herein contain one deuterium atom. In another embodiment, the compounds disclosed herein contain two deuterium atoms. In yet another embodiment, the compounds disclosed herein contain three deuterium atoms. In yet another embodiment, the compounds disclosed herein contain four deuterium atoms. In yet another embodiment, the compounds disclosed herein contain five deuterium atoms. In yet another embodiment, the compounds disclosed herein contain six deuterium atoms. In yet another embodiment, the compounds disclosed herein contain more than six deuterium atoms. In yet another embodiment, the compounds disclosed herein are completely substituted with deuterium atoms and do not contain non-exchangeable ¹ H hydrogen atoms. In one embodiment, the level of deuterium incorporation is determined by the synthesis method using deuterated synthetic building blocks as starting materials.

"Pharmaceutically acceptable salts" include acid addition salts and base addition salts. Any pharmaceutically acceptable salt of the kallikrein inhibitory compounds described herein is intended to include any and all pharmaceutically suitable salt forms. Preferred pharmaceutically acceptable salts of the compounds described herein are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

"Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, etc., which are not biologically or otherwise undesirable and retain the biological effectiveness and properties of the free bases. Also included are salts formed with organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids (including, for example, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.). Thus, exemplary salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, octanoate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, malate, tartrate, methanesulfonate, etc. Salts of amino acids such as arginate, gluconate, and galacturonate are also contemplated (see, e.g., Berge S.M. et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 66: 1-19 (1997)). In some embodiments, acid addition salts of basic compounds are prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt according to methods and techniques well known to those skilled in the art.

"Pharmaceutically acceptable base addition salts" refers to salts that are not biologically or otherwise undesirable and that retain the biological effectiveness and properties of the free acids. These salts are prepared by adding inorganic or organic bases to the free acids. In some embodiments, pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali metals and alkaline earth metals or organic amines. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, tertiary, substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, N,N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenediphenylamine, N-methylglucamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. See Berge et al., supra.

As used herein, "treat" or "treating" or "alleviate" or "relieve" are used interchangeably. These terms refer to an approach for obtaining beneficial or desired results, including but not limited to therapeutic benefit and/or prophylactic benefit. "Therapeutic benefit" means eradication or alleviation of the underlying condition being treated. Additionally, therapeutic benefit can be achieved by eradication or alleviation of one or more physiological symptoms associated with the underlying condition, such that an improvement is observed in the patient, even though the patient still suffers from the underlying condition. For prophylactic benefit, in some embodiments, the composition is administered to a patient at risk for developing a particular disease, or to a patient reporting one or more physiological symptoms of a disease, even though a diagnosis of the disease has not yet been made.

"Prodrug" is intended to mean a compound that, in some embodiments, is converted to a biologically active compound described herein under physiological conditions or by solvolysis. Thus, the term "prodrug" refers to a precursor of a pharmaceutically acceptable biologically active compound. Prodrugs are generally inactive when administered to a subject, but are converted to active compounds in vivo, for example, by hydrolysis. Prodrug compounds generally provide advantages in solubility, tissue compatibility, or sustained release in mammalian organisms (see, e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)).

A discussion of prodrugs is provided in Higuchi, T. et al., "Pro-drugs as Novel Delivery Systems," A.C.S. Symposium Series, Vol. 14, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.

The term "prodrug" is also intended to include any covalently bonded carriers that release the active compound in vivo when administered to a mammalian subject. Prodrugs of the active compounds described herein are prepared by modifying functional groups present in the active compound such that the modifications are cleaved, during routine manipulation or in vivo, to the parent active compound. Prodrugs include compounds in which a hydroxyl, amino, or sulfhydryl group is bonded to any group that, when the prodrug of the active compound is administered to a mammalian subject, cleaves to form a free hydroxyl, free amino, or free sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of alcohol or amine functional groups in the active compound.

Kallikrein inhibitory compounds

Provided herein are heterocyclic derivative compounds and pharmaceutical compositions comprising the same.The subject compounds and compositions are useful for inhibiting plasma kallikrein.

One embodiment provides a compound, or a pharmaceutically acceptable salt thereof, having the structure of Formula (I):

in,

Ring A is an optionally substituted bicyclic heterocycle or bicyclic heteroaryl ring;

Ring B is an optionally substituted monocyclic heterocycle or monocyclic heteroaryl ring, or an optionally substituted bicyclic heterocycle or bicyclic heteroaryl ring;

each R ¹² , R ¹³ or R ¹⁴ is independently selected from hydrogen, cyano, halo, hydroxy, azido, amino, nitro, —CO ₂ H, —S(O)—R ²⁰ , —SR ²⁰ , —S(O) ₂ —R ²⁰ , optionally substituted alkoxy, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted (heterocyclyl)-O—, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted alkylamino, optionally substituted dialkylamino, —CO—R ²⁰ , —CO ₂ —R ²⁰ , —CO(NR ²¹ ) ₂ , —SO ₂ (NR ²¹ ) ₂ , —C(═NR ²² )—(NR ²¹ ) ₂ , or optionally substituted alkynyl;

each R ¹ or R ² is independently selected from hydrogen, halo, hydroxy, amino, -CO ₂ H, -S(O)-R ²⁰ , -SR ²⁰ , -S(O) ₂ -R ^{20 , optionally substituted alkoxy, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted (heterocyclyl)-O-, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted alkylamino, optionally substituted dialkylamino, -CO-R 20 , -CO 2 -R 20} , -CO(NR 21 ) 2 , -SO 2 (NR 21 ) 2 , -C(═NR 22 )-(NR 21 ) 2 or optionally substituted alkynyl; or optionally, R 1 and R 2 are independently selected from hydrogen, halo, hydroxy, amino, -CO 2 H, -S(O)-R ^{20 , -SR 20 , -S(O) 2 -R 20 , optionally substituted alkoxy, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted (heterocyclyl)-O-, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted alkylamino, optionally substituted dialkylamino, -CO-R 20} , -CO ₂ -R ²⁰ , -CO(NR ²¹ ) ₂ , -SO ₂ (NR ²¹ ) ₂ , -C(═NR ²² )-(NR ²¹ ) ₂ or optionally substituted alkynyl ^{; R 2} is an optionally substituted C1-C5 alkyl group and is linked together to form a ring; or optionally, R ¹ and R ² together form an oxo group;

each R ³ or R ⁴ is independently selected from hydrogen, -CO ₂ H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, -CO-R ²⁰ , -CO ₂ -R ²⁰ , -CO(NR ²¹ ) ₂ , -SO ₂ (NR ²¹ ) ₂ , -C(＝NR ²² )-(NR ²¹ ) ₂ or optionally substituted alkynyl; or optionally, R ³ and R ⁴ are optionally substituted C1-C5 alkyl and are linked to form a ring;

each R ²⁰ is selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl;

each R ²¹ is selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl; and

each R ²² is selected from hydrogen, -CN, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclyl; with the proviso that the compound of formula (I) is not 2-[[4,5,6,7-tetrahydro-3-(trifluoromethyl)-1H-indazol-1-yl]methyl]-N-(2-thienylmethyl)-4-pyridinecarboxamide.

Another embodiment provides a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof, wherein Ring A is an optionally substituted bicyclic heterocycle.

Another embodiment provides a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof, wherein Ring A is an optionally substituted bicyclic heteroaryl ring.

Another embodiment provides a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof, wherein Ring B is an optionally substituted monocyclic heterocycle.

Another embodiment provides a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof, wherein Ring B is an optionally substituted monocyclic heteroaryl ring.

Another embodiment provides a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof, wherein Ring B is an optionally substituted bicyclic heterocycle.

Another embodiment provides a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof, wherein Ring B is an optionally substituted bicyclic heteroaryl ring.

Another embodiment provides a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹² is hydrogen.

Another embodiment provides a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹⁴ is independently selected from -S(O)-R ²⁰ , -SR ²⁰ , -S(O) ₂ -R ²⁰ , optionally substituted alkoxy, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted (heterocyclyl)-O-, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted alkylamino, optionally substituted dialkylamino, -CO-R ²⁰ , -CO ₂ -R ²⁰ , -CO(NR ²¹ ) ₂ , -SO ₂ (NR ²¹ ) ₂ , or -C(＝NR ²² )-(NR ²¹ ) ₂ . Another embodiment provides a compound having a structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹⁴ is independently selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl or optionally substituted alkynyl. Another embodiment provides a compound having a structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹⁴ is hydrogen. Another embodiment provides a compound having a structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹⁴ is optionally substituted alkyl or optionally substituted cycloalkyl.

Another embodiment provides a compound having the structure of Formula (I) or a pharmaceutically acceptable salt thereof, wherein R ₁₃ is independently selected from -S(O)-R ²⁰ , -SR ²⁰ , -S(O) ₂ -R ²⁰ , optionally substituted alkoxy, optionally substituted aryloxy, optionally substituted heteroaryloxy, optionally substituted (heterocyclyl)-O-, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted alkylamino, optionally substituted dialkylamino, -CO-R ²⁰ , -CO ₂ -R ²⁰ , -CO(NR ²¹ ) ₂ , -SO ₂ (NR ²¹ ) ₂ , or -C(＝NR ²² )-(NR ²¹ ) ₂ . Another embodiment provides a compound having a structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹³ is independently selected from optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl or optionally substituted alkynyl. Another embodiment provides a compound having a structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹³ is hydrogen. Another embodiment provides a compound having a structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹³ is optionally substituted alkyl or optionally substituted cycloalkyl.

Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ³ and R ⁴ are hydrogen. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ³ is hydrogen. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ⁴ is hydrogen. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ³ is optionally substituted alkyl. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ⁴ is optionally substituted alkyl.

Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹ and R ² are hydrogen. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹ is hydrogen. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ² is hydrogen. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹ is optionally substituted alkyl. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ² is optionally substituted alkyl. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ¹ is optionally substituted alkoxy. Another embodiment provides a compound having the structure of formula (I) or a pharmaceutically acceptable salt thereof, wherein R ² is optionally substituted alkoxy.

One embodiment provides a compound, or a pharmaceutically acceptable salt thereof, having the structure of Formula (Ia):

in,

Ring A is an optionally substituted bicyclic heterocycle or bicyclic heteroaryl ring; and

Ring B is an optionally substituted monocyclic heterocycle or monocyclic heteroaryl ring or an optionally substituted bicyclic heterocycle or bicyclic heteroaryl ring; provided that the compound of formula (I) is not 2-[[4,5,6,7-tetrahydro-3-(trifluoromethyl)-1H-indazol-1-yl]methyl]-N-(2-thienylmethyl)-4-pyridinecarboxamide.

Another embodiment provides a compound having the structure of Formula (Ia) or a pharmaceutically acceptable salt thereof, wherein Ring A is an optionally substituted bicyclic heterocycle.

Another embodiment provides a compound having the structure of Formula (Ia) or a pharmaceutically acceptable salt thereof, wherein Ring A is an optionally substituted bicyclic heteroaryl ring.

Another embodiment provides a compound having the structure of Formula (Ia) or a pharmaceutically acceptable salt thereof, wherein Ring B is an optionally substituted monocyclic heterocycle.

Another embodiment provides a compound having the structure of Formula (Ia) or a pharmaceutically acceptable salt thereof, wherein Ring B is an optionally substituted monocyclic heteroaryl ring.

Another embodiment provides a compound having the structure of Formula (Ia) or a pharmaceutically acceptable salt thereof, wherein Ring B is an optionally substituted bicyclic heterocycle.

Another embodiment provides a compound having the structure of Formula (Ia) or a pharmaceutically acceptable salt thereof, wherein Ring B is an optionally substituted bicyclic heteroaryl ring.

Another embodiment provides a compound having the structure of Formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein Ring B is not thienyl.

Another embodiment provides a compound having the structure of Formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein Ring A is not tetrahydro-1H-indazol-1-yl.

Another embodiment provides a compound having a structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein ring A is selected from optionally substituted quinolinyl, optionally substituted indolyl, optionally substituted indazolyl, optionally substituted benzimidazolyl, optionally substituted isoquinolinyl, optionally substituted cinnolinyl, optionally substituted phthalazinyl, optionally substituted quinazolinyl, optionally substituted naphthyridinyl or optionally substituted benzisoxazolyl. Another embodiment provides a compound having a structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein ring A is selected from optionally substituted benzo[d]isoxazol-7-yl, optionally substituted 4-aminoquinazolin-5-yl, optionally substituted indol-5-yl; optionally substituted quinolin-3-yl; quinoxalin-2-yl; optionally substituted isoquinolin-1(2H)-on-2-yl; or optionally substituted quinolin-6-yl. Another embodiment provides a compound having the structure of Formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein Ring A is an optionally substituted quinolin-6-yl group. Another embodiment provides a compound having the structure of Formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted quinolin-6-yl group is substituted with at least one substituent selected from optionally substituted C1-C3 alkyl, halogen, -CN, _-SO2Me , _-SO2NH2 , _-CONH2 , _-CH2NHAc , _-CO2Me , _-CO2H , _-CH2OH , _-CH2NH2 , _-NH2 , _-OH , or -OMe. Another embodiment provides a compound having the structure of Formula (I) or (Ia), or a pharmaceutically acceptable salt thereof _, wherein the optionally substituted quinolin-6-yl group is substituted at least at the 3-position. Another embodiment provides a compound having the structure of Formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the quinolin-6-yl group is selected from 3-chloroquinolin-6-yl, 3-methylquinolin-6-yl, 3-trifluoromethylquinolin-6-yl, 3-fluoroquinolin-6-yl, or 3-cyanoquinolin-6-yl. Another embodiment provides a compound having the structure of Formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein Ring A is an optionally substituted quinolin-3-yl group. Another embodiment provides a compound having the structure of Formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted quinolin-3-yl group is substituted at least at the 6- or 7-position. Another embodiment provides a compound having the structure of Formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted quinolin-3-yl is substituted with at least one _{substituent selected from optionally substituted C1-C3 alkyl, halogen, -CN, -SO2Me, -SO2NH2, -CONH2, -CH2NHAc, -CO2Me, -CO2H, -CH2OH , -CH2NH2 , -NH2 , -OH , or -OMe .}

Another embodiment provides a compound having the structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein ring B is selected from an optionally substituted monocyclic heteroaryl ring. Another embodiment provides a compound having the structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted monocyclic heteroaryl ring is selected from an optionally substituted imidazolyl, an optionally substituted pyrazolyl, an optionally substituted pyridinyl, an optionally substituted pyridazinyl, an optionally substituted pyrimidinyl, or an optionally substituted pyrazinyl. Another embodiment provides a compound having the structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted monocyclic heteroaryl ring is an optionally substituted pyridinyl. Another embodiment provides a compound having the structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted pyridinyl is an optionally substituted aminopyridinyl. Another embodiment provides a compound having the structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted aminopyridinyl is an optionally substituted 6-aminopyridin-3-yl. Another embodiment provides a compound having a structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein ring B is selected from an optionally substituted bicyclic heteroaryl ring. Another embodiment provides a compound having a structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted bicyclic heteroaryl ring is selected from an optionally substituted quinolyl, an optionally substituted isoquinolyl, an optionally substituted quinazolinyl, an optionally substituted indolyl, an optionally substituted indazolyl, an optionally substituted 1H-pyrrolo[2,3-b]pyridinyl, an optionally substituted benzoxazolyl, an optionally substituted benzisoxazolyl, or an optionally substituted benzimidazolyl. Another embodiment provides a compound having a structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted bicyclic heteroaryl ring is an optionally substituted indolyl. Another embodiment provides a compound having a structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted bicyclic heteroaryl ring is an optionally substituted indazolyl. Another embodiment provides a compound having the structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted indolyl is an optionally substituted indol-5-yl. Another embodiment provides a compound having the structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted indazolyl is an optionally substituted indazol-5-yl. Another embodiment provides a compound having the structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein the optionally substituted bicyclic heteroaryl ring is an optionally substituted 1H-pyrrolo[2,3-b]pyridinyl. Another embodiment provides a compound having the structure of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof, wherein Ring A is selected from optionally substituted quinolinyl; and Ring B is selected from optionally substituted indolyl, optionally substituted indazolyl, and optionally substituted 1H-pyrrolo[2,3-b]pyridinyl.

In some embodiments, the kallikrein-inhibiting compound described in Formula (I) has the structure provided in Table 1.

Table 1

In some embodiments, the compounds described herein have the structures provided in Table 2.

Table 2

Preparation of compounds

The compounds used in the reactions described herein are prepared according to organic synthesis techniques known to those skilled in the art, starting from commercially available chemicals and/or compounds described in the chemical literature. “Commercially available chemicals” were obtained from standard commercial sources, including Acros Organics (Pittsburgh, PA), Aldrich Chemical (Milwaukee, WI, including Sigma Chemical and Fluka), Apin Chemicals Ltd. (Milton Park, U.K.), Avocado Research (Lancashire, U.K.), BDH Inc. (Toronto, Canada), Bionet (Cornwall, U.K.), Chemservice Inc. (West Chester, PA), Crescent Chemical Co. (Hauppauge, NY), Eastman Organic Chemicals, Eastman Kodak Company (Rochester, NY), Fisher Scientific Co. (Pittsburgh, PA), Fisons Chemicals (Leicestershire, U.K.), Frontier Scientific (Logan, UT), ICN Biomedicals, Inc. (Costa Mesa, CA), Key Organics (Cornwall, U.K.), Lancaster Synthesis (Windham, NH), Maybridge Chemical Co. Ltd. (Cornwall, U.K.), Parish Chemical Co. (Orem, UT), Pfaltz & Bauer, Inc. (Waterbury, CN), Polyorganix (Houston, TX), Pierce Chemical Co. (Rockford, IL), Riedel de Haen AG (Hanover, Germany), Spectrum Quality Product, Inc. (New Brunswick, NJ), TCI America (Portland, OR), Trans World Chemicals, Inc. (Rockville, MD), and Wako Chemicals USA, Inc. (Richmond, VA).

Suitable reference books and treatises that describe in detail the synthesis of reactants useful in the preparation of the compounds described herein, or provide literature references describing such preparations, include, for example, "Synthetic Organic Chemistry", John Wiley & Sons, Inc., New York; S. R. Sandler et al., "Organic Functional Group Preparations," 2nd ed., Academic Press, New York, 1983; H. O. House, "Modern Synthetic Reactions," 2nd ed., W. A. Benjamin, Inc. Menlo Park, Calif. 1972; T. L. Gilchrist, "Heterocyclic Chemistry," 2nd ed., John Wiley & Sons, New York, 1992; J. March, "Advanced Organic Chemistry: Reactions, Mechanisms and Structure," 4th ed., Wiley-Interscience, New York, 1992. Other suitable reference books and treatises that describe in detail the synthesis of reactants useful in the preparation of the compounds described herein or provide literature references describing such preparations include, for example, Fuhrhop, J. and Penzlin G. "Organic Synthesis: Concepts, Methods, Starting Materials", 2nd ed., Revised and Supplemented (1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R.V. "Organic Chemistry, An Intermediate Text" (1996) Oxford University Press, ISBN 0-19-509618-5; Larock, R.C. "Comprehensive Organic Transformations: A Guide to Functional Group Preparations", 2nd ed. (1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J. "Advanced Organic Chemistry: Reactions, Mechanisms, and Structure", 4th ed. (1992) John Wiley & Sons, ISBN: 0-471-60180-2; Otera, J. (editor) "Modern Carbonyl Chemistry" (2000) Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. "Patai's 1992 Guide to the Chemistry of Functional Groups" (1992) Interscience ISBN: 0-471-93022-9; Solomons, T.W.G. "Organic Chemistry" Seventh Edition (2000) John Wiley & Sons, ISBN: 0-471-19095-0; Stowell, J.C., "Intermediate Organic" Chemistry" Second Edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2; "Industrial Organic Chemicals: Starting Materials and "Intermediates: An Ullmann's Encyclopedia" (1999) John Wiley & Sons, ISBN: 3-527-29645-X, a total of 8 volumes; "Organic Reactions" (1942-2000) John Wiley & Sons, a total of more than 55 volumes; and "Chemistry of Functional Groups" John Wiley & Sons, a total of 73 volumes.

Specific and similar reactants are optionally identified by indexing known chemicals compiled by the Chemical Abstracts Service of the American Chemical Society, which is available in most public and university libraries and through online databases (contact the American Chemical Society, Washington, D.C. for more details). Chemicals known in the catalog but not commercially available are optionally prepared by custom chemical synthesis facilities, many of which provide custom synthesis services. A reference for the preparation and selection of pharmaceutically acceptable salts of the kallikrein inhibitory compounds described herein is P.H. Stahl and C.G. Wermuth, "Handbook of Pharmaceutical Salts", Verlag Helvetica Chimica Acta, Zurich, 2002.

Pharmaceutical composition

In certain embodiments, the kallikrein inhibitory compounds described herein are administered as pure chemicals. In other embodiments, the kallikrein inhibitory compounds described herein are combined with a pharmaceutically suitable or acceptable carrier (also referred to herein as a pharmaceutically suitable (or acceptable) excipient, a physiologically suitable (or acceptable) excipient, or a physiologically suitable (or acceptable) carrier), the selection of which is based on the selected route of administration and standard pharmaceutical practice as described, for example, in Remington: The Science and Practice of Pharmacy (Gennaro, 21st ed. Mack Pub. Co., Easton, PA (2005)).

Provided herein is a pharmaceutical composition comprising at least one kallikrein inhibitory compound, or a stereoisomer, pharmaceutically acceptable salt, hydrate, solvate, or N-oxide thereof, and one or more pharmaceutically acceptable carriers. A carrier (or excipient) is acceptable or suitable if it is compatible with the other ingredients of the composition and is not harmful to the recipient (i.e., subject) of the composition.

One embodiment provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. One embodiment provides a pharmaceutical composition comprising a compound of formula (Ia) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

In certain embodiments, the kallikrein inhibitory compound as described in Formula (I) or (Ia) is substantially pure in that it contains less than about 5%, or less than about 1%, or less than about 0.1% of other small organic molecules, such as unreacted intermediates or synthesis byproducts produced, for example, during one or more steps of the synthetic method.

Suitable oral dosage forms include, for example, tablets, pills, capsules, or capsules of hard or soft gelatin, methylcellulose, or another suitable material that dissolves readily in the digestive tract. In some embodiments, suitable non-toxic solid carriers are used, including, for example, pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, magnesium carbonate, etc. (see, for example, Remington: The Science and Practice of Pharmacy (Gennaro, 21st ed. Mack Pub. Co., Easton, PA (2005)).

The dosage of the composition comprising at least one kallikrein-inhibiting compound as described herein varies depending on the condition of the patient (eg, human), ie, disease stage, general health, age, and other factors.

The pharmaceutical composition is applied in a manner suitable for the disease to be treated (or prevented). Appropriate dosage and suitable administration duration and frequency will depend on factors such as the patient's condition, the type and severity of the patient's disease, the concrete form of the active ingredient and the method of administration. Typically, the amount of the composition provided by appropriate dosage and treatment regimen is enough to provide treatment and/or prevention benefit (for example, improved clinical outcome, such as more frequent complete or partial remission, or longer disease-free and/or total survival period, or symptom severity alleviation). Optimal dose is typically determined using experimental models and/or clinical trials. Optimal dose depends on the patient's body mass, body weight or blood volume.

Oral dosages generally range from about 1.0 mg to about 1000 mg, 1 to 4 times a day or more.

Kallikrein-kinin system

Regulation of vascular permeability is crucial in regulating the passage of small molecules or blood cells between blood vessels and surrounding tissues. Vascular permeability depends on the physiological state of the tissue, changes in blood pressure, and fluctuations in ion and nutrient gradients, such as during inflammation. The junctions between the endothelial cells lining the blood vessels are the direct control factors of vascular permeability. The strength of these junctions is strictly regulated by the kinin-kallikrein system of polypeptides and enzymes. Abnormalities in the kinin-kallikrein system lead to a series of pathologies, including angioedema, macular edema, and cerebral edema. Angioedema is a potentially fatal blood disorder characterized by swelling that may occur in the face, gastrointestinal tract, limbs, genitals, and upper respiratory tract. Hereditary hereditary angioedema attacks are caused by unregulated activation of the kallikrein system, accompanied by uncontrolled increased vascular permeability. There is currently a need for agents that can be used to treat angioedema and agents that inhibit plasma kallikrein.

The kallikrein-kinin system represents a metabolic cascade that, when activated, triggers the release of vasoactive kinins. The kinin-kallikrein system (KKS) is composed of serine proteases involved in the production of kinins, primarily bradykinin and lysine-bradykinin (kallikrein). KKS contributes to a variety of physiological processes, including inflammation, blood pressure control, and coagulation. Due to the ability of bradykinin to increase vascular permeability and cause vasodilation of arteries and veins in the intestine, aorta, uterus, and urethra, activation of this system is particularly important in blood pressure regulation and inflammatory responses. The kinin-kallikrein system, also known as the contact system, consists of three serine zymogens (factor XII (FXII) or Hageman factor, factor IX (FIX), and prekallikrein) and the kinin precursor high molecular weight kinin (HK). Contact activation is triggered by the binding of FXII to a negatively charged surface and involves the formation of α-FXIIa via an autocatalytic reaction. The bound α-FXIIa converts prekallikrein into kallikrein. Kallikrein can further convert α-FXIIa to β-FXIIa through an additional cleavage at R334-N335, a positive feedback mechanism that results in sufficient kallikrein production to drive downstream processes. α-FXIIa consists of a heavy chain and a light chain linked by a disulfide bond, while β-FXIIa lacks the heavy chain and loses its ability to bind to negatively charged surfaces (Stavrou E, Schmaier AH., Thrombosis Research, 2010, 125(3)pp.210-215). The N-terminal region of FXII (α-FXIIa heavy chain) shows strong homology with tissue plasminogen activator (tPA), and has type I fibronectin, epidermal growth factor, and Kringle domains (Ny et al., Proc Natl Acad Sci U S A, 1984, 81 (17) pp. 5355-5359; Cool DE, MacGillivray RT, The Journal of Biological Chemistry, 1987, 262 (28) pp. 13662-13673). Kallikrein is a trypsin-like serine protease that cleaves high molecular weight kinin (HK) to produce bradykinin. Bradykinin then binds to the bradykinin 2R receptor (BK2R) on endothelial cells to trigger an increase in vascular permeability.

Protease inhibitors regulate the activation of this contact system. Several known serine protease inhibitors (serpins) of plasma are C1-inhibitor (C1INH), antithrombin III, α2-macroglobulin, α1-proteinase inhibitor, and α2-antiplasmin (Kaplan et al., Advances in Immunology, 1997 (66) pp. 225-72; Pixley et al., The Journal of Biological Chemistry, 1985, 260 (3) pp. 1723-9). However, C1INH is the main regulator of the intrinsic system, interfering with the activity of factor XIIa and kallikrein (Cugno et al., The Journal of Laboratory and Clinical Medicine, 1993, 121 (1) pp. 38-43). C1INH and α2-macroglobulin together account for 90% of the kallikrein inhibitory activity of plasma. Thus, the FXII-dependent kallikrein-kinin system is tightly regulated by CINH, and when the regulation of the FXII-dependent kallikrein-kinin system fails in a subject, the subject is considered to suffer from hereditary angioedema (HAE), which is characterized by invalidating episodes of edema.

Angioedema is a potentially fatal blood disorder characterized by swelling that may occur in the face, gastrointestinal tract, extremities, genitals, and upper respiratory tract. Angioedema attacks begin in the deeper layers of the skin and mucous membranes with localized vasodilation and increased permeability. The symptoms of the disease are caused by the leakage of plasma from blood vessels into surrounding tissues. Hereditary angioedema attacks result from unregulated activation of the kallikrein system, with subsequent overproduction of bradykinin and uncontrolled increased vascular permeability. When vascular permeability increases beyond normal levels, plasma leaks from the vasculature into the surrounding tissues, causing swelling (Mehta D and Malik AB, Physiol. Rev., 86(1), 279-367, 2006; Sandoval R et al., J. Physiol., 533(pt 2), 433-45, 2001; Kaplan AP and Greaves MW, Angioedema. J. Am. Acad. Dermatol., 2005).

HAE is caused by mutations in genes encoding elements of the coagulation and inflammatory pathways. The three forms of HAE are distinguished by their underlying cause and the level of C1-esterase inhibitor (C1INH, serpin peptidase inhibitor, branch G, member 1) protein in the blood, which inhibits the activity of plasma kallikrein. In type I, patients have insufficient functional C1INH levels, while type II patients have dysfunctional C1INH. Types I and II affect men and women in equal proportions, while type III primarily affects women and is caused by mutations in coagulation factor XII (Hageman factor; HAE-FXII). The underlying cause of type I and type II HAE is an autosomal dominant mutation in the C1INH gene (SERPING1 gene) on chromosome 11 (11q12-q13.1).

C1INH accounts for 90% of FXIIa inhibition and 50% of plasma kallikrein inhibition (Pixley RA et al., J. Biol. Chem., 260, 1723–9, 1985; Schapira M et al., Biochemistry, 20, 2738–43, 1981). In addition, C1INH also inactivates prekallikrein (Colman RW et al., Blood, 65, 311-8, 1985). When C1INH levels are normal, its activity blocks FXIIa from converting prekallikrein to kallikrein and blocks the conversion of kallikrein to HK, thereby preventing bradykinin production and the onset of edema. When C1INH levels are low, or when levels of dysfunctional C1INH are high, this inhibition fails and the pathogenic process ensues.

In addition to HAE, plasma kallikrein also causes non-hereditary angioedema, high altitude cerebral edema, cytotoxic cerebral edema, osmotic cerebral edema, diabetic macular edema (DME), clinically significant macular edema, cystoid macular edema (CME, Gao BB, Nat Med., 13(2), 181-8, 2007), retinal edema, radiation-induced edema, lymphedema, glioma-related edema, allergic edema, such as airway obstruction in chronic allergic sinusitis or perennial rhinitis. Other disorders of the plasma kallikrein system include retinopathy and diabetic retinopathy (Liu J and Feener EP, Biol. Chem. 394(3), 319-28, 2013), proliferative and nonproliferative retinopathy (Liu J et al., Invest. Ophthalmol. Vis. Sci., 54(2), 2013), CME after cataract extraction, cryotherapy-induced CME, uveitis-induced CME, CME after vascular occlusion (e.g., central retinal vein occlusion, branch retinal vein occlusion, or hemiretinal vein occlusion), complications of diabetic retinopathy associated with cataract surgery, hypertensive retinopathy (JA Phillips et al., Hypertension, 53, 175-181, 2009), retinal trauma, dry and wet age-related macular degeneration (AMD), ischemia-reperfusion injury (C Storoni et al., JPET, 381, 849-954, 2006), for example, in various situations related to tissue and/or organ transplantation.

Current treatments for angioedema, and those in development, target different elements in the HAE pathway. Three classes of therapy are currently available: (a) replacement therapy using C1INH concentrates (e.g., Cinryze, Berinert), (b) administration of selective kallikrein inhibitors (e.g., Ecallantide), and (c) bradykinin receptor antagonists (e.g., Firazyr).

Alternative therapy has been shown to be useful for acute attacks, including emergency situations such as laryngeal edema (Bork K et al., Transfusion, 45, 1774–1784, 2005; Bork K and Barnstedt SE, Arch. Intern. Med., 161, 714–718, 2001), as well as for prevention. Selective C1INH inhibitors inactivate both α-FXIIa and β-FXIIa molecules that are active early in the HAE pathway and catalyze the production of kallikrein (Muller F and Renne T, Curr. Opin. Hematol., 15, 516–21, 2008; Cugno M et al., Trends Mol. Med. 15(2): 69-78, 2009). In addition to HAE, plasma kallikrein inhibitors are also considered useful for treating other edemas, such as macular edema and cerebral edema, as well as retinal diseases, for example, those associated with diabetes and/or hypertension. There is evidence that plasma kallikrein inhibitors are also effective in treating edema in diseases such as ischemia-reperfusion injury. Bradykinin receptor antagonists prevent bradykinin from activating the vascular permeability pathway and halt the initiation of swelling.

Treatment

Disclosed herein are methods of treating diseases or conditions in which inhibition of plasma kallikrein is shown. Such diseases or conditions include, but are not limited to, angioedema, including hereditary and non-hereditary angioedema.

In some embodiments, the methods disclosed herein can be used to treat angioedema. In some embodiments, the angioedema is hereditary angioedema (HAE). One embodiment provides a method for treating angioedema in a patient in need thereof, comprising administering a composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. Another embodiment provides the method, wherein the angioedema is hereditary angioedema.

One embodiment provides a method of treating angioedema in a patient in need thereof, comprising administering a composition comprising a compound of Formula (Ia) or a pharmaceutically acceptable salt thereof. Another embodiment provides the method, wherein the angioedema is hereditary angioedema.

Other embodiments and uses will be apparent to those skilled in the art in light of this disclosure.The following examples are provided merely as illustrations of various embodiments and should not be construed in any way as limiting the invention.

Example

I. Chemical Synthesis

Unless otherwise stated, reagents and solvents were used as received from commercial suppliers. Anhydrous solvents and oven-dried glassware were used for synthetic transformations that were sensitive to moisture and/or oxygen. Yields were not optimized. Reaction times are approximate and not optimized. Unless otherwise stated, column chromatography and thin layer chromatography (TLC) were performed on silica gel. Spectra are given in ppm (δ), and coupling constants J are reported in Hertz. For proton spectra, the solvent peak was used as the reference peak.

The following abbreviations and terms have the indicated meanings throughout the text:

AcOH = acetic acid

B ₂ pin ₂ = bis(pinacol)diboron

Boc = tert-butyloxycarbonyl

DCC = dicyclohexylcarbodiimide

DIEA＝N,N-diisopropylethylamine

DMAP = 4-dimethylaminopyridine

EDC = 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide

eq = equivalent

Et = Ethyl

EtOAc or EA = ethyl acetate

EtOH = ethanol

g = grams

h or hr = hours

HBTU = O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate

HOBt = Hydroxybenzotriazole

HPLC = High Pressure Liquid Chromatography

kg or Kg = kilogram

L or l = liter

LC/MS = Liquid Chromatography-Mass Spectrometry

LRMS = low-resolution mass spectrometry

m/z = mass-to-charge ratio

Me = methyl

MeOH = methanol

mg = milligrams

min = minutes

mL = milliliters

mmol = millimole

NaOAc = sodium acetate

PE = Petroleum Ether

Ph = phenyl

Prep＝Preparative

quant.＝quantitative

RP-HPLC = Reverse Phase High Pressure Liquid Chromatography

rt or RT = room temperature

THF = Tetrahydrofuran

UV = Ultraviolet

Intermediate 1: Preparation of 2-((2-cyanoquinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of 6-methylquinoline 1-oxide

To a solution of 6-methylquinoline (10.0 g, 69.9 mmol, 1.0 equiv) in HOAc (150 mL) was added 30% _H2O2 (100 _mL ). The reaction mixture was heated to 70°C and stirred overnight, then cooled, water (100 mL) was _added , and then _Na2SO3 was added portionwise while cooling in an ice bath to quench _the excess _H2O2 . The reaction mixture was extracted with DCM, and the organic phase was washed with water, brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to give 6-methylquinoline 1-oxide (7.2 g, 64%) as a brown oil.

Step 2: Preparation of 6-methylquinoline-2-carbonitrile

While cooling in an ice-water bath, to a solution of 6-methylquinoline 1-oxide (7.2 g, 45.2 mmol, 1.0 equiv) in trimethylsilyl cyanide (17.0 mL, 135.8 mmol, 3.0 equiv) was added benzoyl chloride (15.6 mL, 135.8 mmol, 3.0 equiv) followed by triethylamine (18.9 mL, 135.8 mmol, 3.0 equiv). The reaction was stirred for 1 h. The mixture was diluted with DCM and carefully washed once with saturated aqueous NaHCO ₃ solution, then with water, brine, dried over anhydrous sodium sulfate, and purified on a silica gel column (PE/DCM/EtOAc = 10/1/1) followed by trituration with EtOH to give 6-methylquinoline-2-carbonitrile (6.0 g, 78%) as a yellow solid.

Step 3: Preparation of 6-(bromomethyl)quinoline-2-carbonitrile

A mixture of 6-methylquinoline-2-carbonitrile (3.7 g, 22.0 mmol, 1.0 eq), NBS (3.9 g, 22.0 mmol, 1.0 eq) and AIBN (72 mg, 2 mol%) in carbon tetrachloride (100 mL) was refluxed for 3 h, then cooled and concentrated to dryness. The residue was triturated with DCM, filtered and dried to give 6-(bromomethyl)quinoline-2-carbonitrile (4.0 g, 74%) as a white solid.

Step 4: Preparation of ethyl 2-((2-cyanoquinolin-6-yl)methyl)isonicotinate

A mixture of ethyl 2-bromoisonicotinate (1.86 g, 8.1 mmol, 1.0 equiv), BPDB (2.06 g, 8.1 mmol, 1.0 equiv), potassium acetate (2.38 g, 24.3 mmol, 3.0 equiv), and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (296 mg, 5 mol%) was mixed in 1,4-dioxane (100 mL). The mixture was degassed with nitrogen, heated to 85° C. and stirred for 16 h, then cooled to room temperature, and 6-(bromomethyl)quinoline-2-carbonitrile (2.0 g, 8.1 mmol, 1.0 equiv), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (296 mg, 5 mol%), and sodium carbonate (2.57 g, 24.3 mmol, 3.0 equiv, dissolved in 30 mL of water) were added. The mixture was degassed, heated to 95 ° C and stirred overnight. The mixture was cooled and filtered through celite. Water (100 mL) and DCM (100 mL) were added to the filtrate. The DCM layer was separated and washed with brine, dried over anhydrous sodium sulfate, and purified on a silica gel column to give ethyl 2-((2-cyanoquinolin-6-yl)methyl)isonicotinate (670 mg, 26%) as a red solid.

Step 5: Preparation of 2-((2-cyanoquinolin-6-yl)methyl)isonicotinic acid

To a solution of ethyl 2-((2-cyanoquinolin-6-yl)methyl)isonicotinate (670 mg, 2.1 mmol, 1.0 equiv) in THF (6 mL) was added an aqueous solution of lithium hydroxide monohydrate (177 mg, 4.2 mmol, 2.0 equiv) in water (3 mL). The reaction was stirred for 2 h and then acidified to pH 3 with 2N HCl. DCM (50 mL) and water (50 mL) were added, and the DCM layer was separated and washed with brine, dried over anhydrous sodium sulfate, and purified by flash chromatography to give 2-((2-cyanoquinolin-6-yl)methyl)isonicotinoic acid (250 mg, 80% purity) as a brown solid.

Intermediate 2: Preparation of 2-((3-chloroquinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of methyl 3-chloroquinoline-6-carboxylate

To a solution of methyl quinoline-6-carboxylate (15.0 g, 80.2 mmol, 1.0 equiv) in DMF (200 ml) was added N-chlorosuccinimide (21.4 g, 0.16 mol, 2.0 equiv) and the reaction mixture was stirred at 120 ° C for 20 h. The reaction mixture was cooled to room temperature, treated with brine, and the mixture was extracted with ethyl acetate. The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography on silica gel (EtOAc/PE=1/8, v/v) to give methyl 3-chloroquinoline-6-carboxylate (9.1 g, 51%) as a yellow solid.

Step 2: Preparation of methyl (3-chloro-quinolin-6-yl)-methanol

To a solution of 3-chloroquinoline-6-formic acid methyl ester (8g, 36.0mmol, 1.0 equivalent) in anhydrous THF was added LiAlH ₄ (2.5M in THF, 5.8mL, 0.4 equivalent). The resulting mixture was stirred at 0°C for 1h. Thereafter, additional LiAlH ₄ (2.5M in THF, 2.8mL, 0.2 equivalent) was added. The system was stirred for another 30min at 0°C and quenched by slowly adding a 1N NaOH aqueous solution. The resulting precipitate was filtered, and the filtrate was extracted with ethyl acetate. The combined organic layers were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=20/1～5/1, v/v) to obtain (3-chloro-quinoline-6-yl)-methanol (4.8g, 69%) as a white solid.

Step 3: Preparation of 3-chloro-6-chloromethyl-quinoline

To (3-chloro-quinolin-6-yl)-methanol (3.3 g, 17.1 mmol, 1.0 equiv) was added _SOCl₂ (50 mL), and the mixture was stirred at room temperature for 1 h. The volatiles were removed under vacuum and the residue was dissolved in DCM. The mixture was washed with saturated aqueous _NaHCO₃ , dried, and concentrated to give 3-chloro-6-chloromethyl-quinoline (3.4 g, 94%) as a yellow solid.

Step 4: Preparation of methyl 2-(trimethylstannyl)isonicotinate

Hexamethyldistanane (0.21 mL, 334 mg, 1.02 mmol) and tetrakis(triphenylphosphine)palladium(0) (70 mg, 0.06 mmol) were added to a solution of methyl 2-chloroisonicotinate (100 mg, 0.58 mmol) in anhydrous dioxane (10 mL), and the resulting mixture was refluxed under _N for 3 h. EtOAc (50 mL) and water (100 mL) were then added. The layers were separated and the organic layer was washed with water (5×100 mL), dried (Na ₂ SO ₄ ), and the solvent was removed by rotary evaporation to leave a crude residue that was used in the next step without further purification.

Step 4: Preparation of methyl 2-((3-chloroquinolin-6-yl)methyl)isonicotinate

To a solution of 3-chloro-6-chloromethyl-quinoline (110 mg, 0.52 mmol, 1.0 eq) and crude methyl 2-(trimethylstannyl)isonicotinate in dioxane (10 mL) was added Pd(PPh ₃ ) ₂ Cl ₂ (36 mg, 0.05 mmol, 0.1 eq). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, the solvent was removed, and the mixture was purified by silica gel chromatography (EtOAc/PE=10/1 to 5:1, v/v) to give methyl 2-((3-chloroquinolin-6-yl)methyl)isonicotinate (70 mg) as a yellow solid. ¹ HNMR (400MHz, DMSO-d ₆ )δ8.83(m,2H),8.53(d,1H),7.98(d,1H),7.85(d,1H),7.73(dd,1H),7.58(s,1H),7.57(s,1H),4.36(s,2H),3.85(s,3H).

Step 5: Preparation of 2-((3-chloroquinolin-6-yl)methyl)isonicotinic acid

To a solution of methyl 2-((3-chloroquinolin-6-yl)methyl)isonicotinate (70 mg, 0.22 mmol, 1.0 equiv) in THF/ _H₂O (5 mL/1 mL) was added LiOH (71 mg, 2.1 mmol, 10 equiv). The resulting mixture was stirred at room temperature for 1 h; all starting material was consumed (estimated by TLC). The volatile solvents were removed on a rotary evaporator, and the aqueous residue was neutralized with ₁ M HCl and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine, dried over _Na₂SO₄ , and concentrated to provide the crude acid (50 mg, 75%), which was used directly without further purification.

Intermediate 3: Preparation of 2-((3-chloro-8-fluoroquinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of methyl 8-fluoroquinoline-6-carboxylate

A mixture of methyl 4-amino-3-fluorobenzoate (35 g, 0.207 mmol, 1 eq), acrolein (17.4 g, 0.311 mol, 1.5 eq) and 6N HCl (600 mL) was stirred at 100 ° C for 10 min. The mixture was then cooled and adjusted to pH ~ 5-6 with NaHCO ₃ (aq). The mixture was extracted with DCM. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered, then concentrated and purified by column chromatography (EtOAc/PE=1/20, v/v) to give methyl 8-fluoroquinoline-6-carboxylate (11 g, 21%) as a yellow solid.

Step 2: Preparation of methyl 3-chloro-8-fluoroquinoline-6-carboxylate

To a DMF solution of 8-fluoroquinoline-6-methyl formate (11 g, 53.7 mmol, 1 equivalent) was added NCS (21.4 g, 0.161 mol, 3 equivalents). The reaction mixture was stirred at 120 ° C overnight. The reaction mixture was cooled to ambient temperature, treated with water, neutralized with solid NaHCO ₃ , and further stirred at room temperature for 30 min. Finally, powdered sodium thiosulfate was carefully added to remove excess NCS. The mixture was extracted with ethyl acetate. The organic layer was dried and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel to obtain 3-chloro-8-fluoroquinoline-6-methyl formate (11.5 g, 90%) as a yellow solid.

Step 3: Preparation of (3-chloro-8-fluoro-quinolin-6-yl)-methanol

To a solution of 3-chloro-8-fluoroquinoline-6-methyl carboxylate (4.5 g, 18.8 mmol, 1 equivalent) was added LiAlH(t-BuO) ₃ (12.0 g, 47.1 mmol, 2.5 equivalents). The resulting mixture was stirred at 40 ° C for 12 h and then quenched by adding water. The mixture was extracted with ethyl acetate. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=2/1, v/v) to give (3-chloro-8-fluoro-quinoline-6-yl)-methanol (2.1 g, 53%) as a yellow solid.

Step 4: Preparation of 3-chloro-6-chloromethyl-8-fluoro-quinoline

A mixture of 3-chloro-8-fluoro-6-hydroxymethyl-quinoline (2.1 g, 9.95 mmol, 1.0 equiv) in _SOCl2 (50 mL) was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DCM and treated with saturated _NaHCO3 solution to give 3-chloro-6-chloromethyl-8-fluoro-quinoline (2.2 g, 96%) as a yellow solid.

Step 5: Preparation of methyl 2-((3-chloro-8-fluoroquinolin-6-yl)methyl)isonicotinate

To a solution of 3-chloro-6-chloromethyl-8-fluoro-quinoline (2.2 g, 9.61 mmol, 1.0 equiv) in dioxane (60 mL) was added methyl 2-(trimethylstannyl)isonicotinate (3.18 g, 10.6 mmol, 1.1 equiv) and Pd(PPh ₃ ) ₂ Cl ₂ (674 mg, 0.96 mmol, 0.1 equiv). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, concentrated, and purified by silica gel chromatography (DCM/MeOH=200/1, v/v) to give methyl 2-((3-chloro-8-fluoroquinolin-6-yl)methyl)isonicotinate (1.6 g, 50%) as a yellow solid.

Step 6: Preparation of 2-((3-chloro-8-fluoroquinolin-6-yl)methyl)isonicotinic acid

To a solution of methyl 2-((3-chloro-8-fluoroquinolin-6-yl)methyl)isonicotinate (800 mg, 2.4 mmol, 1 eq) in THF (20 ml)/water (10 ml) was added NaOH (116 mg, 0.29 mmol, 1.2 eq). The mixture was stirred at room temperature for 3 h. Aqueous HCl (2N) was then added to the reaction mixture until the pH was 6-7. The mixture was extracted with ethyl acetate, and the organic layer was concentrated under pressure. The gray compound was used directly in the next step (500 mg, 76%).

Intermediate 4: Preparation of 2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinic acid

The title compound was synthesized as described for Intermediate 3 using methyl 4-amino-3-chlorobenzoate as the starting material.

Intermediate 5: Preparation of 2-((4-cyanoquinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of 6-(methoxycarbonyl)quinoline 1-oxide

A mixture of methyl quinoline-6-carboxylate (10 g, 53.5 mmol, 1 eq) and m-CPBA (18.4 g, 0.106 mol, 2 eq) in DCM (50 mL) was stirred at room temperature overnight. Saturated _NaHCO aqueous solution (40 mL) was added to the reaction mixture and stirred for 30 min. The organic layer was separated, dried, filtered, and concentrated to obtain a residue which was recrystallized from ethyl acetate (5 mL) to afford 6-(methoxycarbonyl)quinoline 1-oxide (8.0 g, 74%) as a light yellow solid.

Step 2: Preparation of methyl 2-chloroquinoline-6-carboxylate and methyl 4-chloroquinoline-6-carboxylate

To 6-(methoxycarbonyl)quinoline 1-oxide (4.0 g, 19.7 mmol, 1 equivalent) is added phosphorus oxychloride (20 mL). The resulting mixture is then stirred at room temperature for 2 h under _N . Volatiles are then removed under vacuum and the residue is dissolved in DCM. The mixture is washed with saturated NaHCO ₃ aqueous solution, dried and concentrated. The residue is purified by silica gel chromatography (PE/EtOAc=10/1, v/v) to obtain 2-chloroquinoline-6-methyl formate (1.2 g, 28%) and 4-chloroquinoline-6-methyl formate (2.5 g, 57%).

Step 3: Preparation of (4-chloro-quinolin-6-yl)-methanol

To a solution of methyl 4-chloroquinoline-6-formate (2.2 g, 10 mmol, 1 equivalent) was added LiAlH(t-BuO) ₃ (7.62 g, 30 mmol, 3 equivalents). The resulting mixture was stirred at 60 ° C for 2 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=1/1, v/v) to give (4-chloro-quinoline-6-yl)-methanol (1.54 g, 80%) as a yellow solid.

Step 4: Preparation of 6-hydroxymethyl-quinoline-4-carbonitrile

To a mixture of (4-chloroquinoline-6-yl)methanol and zinc cyanide in DMF (30mL) is added tetrakistriphenylphosphine palladium. The mixture is degassed and then heated to 90 ° C for 3h. Afterwards, the mixture is diluted with water and EtOAc, filtered, and the resulting layer is separated. The aqueous layer is further extracted with EtOAc. The combined extracts are washed with water, dried over _MgSO4 , filtered, and the solvent is removed in vacuo. The resulting residue is purified by chromatography on a silica gel column to obtain 6-hydroxymethyl-quinoline-4-carbonitrile (1.1g, 77%) as an off-white solid.

Step 5: Preparation of 2-((4-cyanoquinolin-6-yl)methyl)isonicotinic acid

The title compound was prepared as described for Intermediate 3, Steps 4, 5 and 6.

Intermediate 6: Preparation of 2-((7-chloroquinolin-3-yl)methyl)isonicotinic acid

Step 1: Preparation of N'-(7-chloroquinoline-3-carbonyl)-4-methylbenzenesulfonylhydrazide

To a solution of ethyl 7-chloro-quinoline-3-carboxylate (3.5 g, 14.90 mmol, 1.0 equivalent) in EtOH (60 mL) was added hydrazine monohydrate (7.2 mL, 149 mmol, 10 equivalents). The reaction mixture was stirred at 80 ° C for 2 h and then concentrated under reduced pressure. Water was added to the reaction flask, the solid was filtered and washed with cold water. The solid was air-dried and then dissolved in pyridine (30 mL). TsCl (3.4 g, 17.90 mmol, 1.2 equivalents) was added to the mixture. After stirring at room temperature for 1 h, the mixture was concentrated in vacuo. The residue was poured into water, and the resulting precipitate was collected by filtration to obtain N'-(7-chloroquinoline-3-carbonyl)-4-methylbenzenesulfonylhydrazide (5.0 g, 90%) as a yellow solid.

Step 2: Preparation of (7-chloro-quinolin-3-yl)-methanol

A mixture of N'-(7-chloroquinoline-3-carbonyl)-4 _- methylbenzenesulfonylhydrazide (5.0 g, 13.30 mmol, 1.0 eq) and _Na2CO3 (4.24 g, 40 mmol, 3 eq) in ethylene glycol (30 mL) was heated at 160°C for 20 min. After cooling to room temperature, the mixture was diluted with water and extracted with _Et2O . The combined extracts were dried and concentrated. The residue was purified by flash chromatography on a silica gel column (PE/EtOAc=1/1, v/v) to give (7-chloro-quinolin-3-yl)-methanol (600 mg, 23%) as a yellow solid.

Step 3: Preparation of 2-((7-chloroquinolin-3-yl)methyl)isonicotinic acid

The title compound was synthesized as described for Intermediate 3, Steps 4, 5 and 6.

Intermediate 7: Preparation of 2-((3-methylisoquinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of 6-bromo-3-methyl-isoquinoline

To a solution of 4-bromo-benzylamine (10.0 g, 54 mmol, 1.0 equivalent) in DCE (100 mL) was added 1,1-dimethoxy-propan-2-one (7.0 g, 59 mmol, 1.1 equivalents) and MgSO ₄ (20 g). The mixture was stirred at 40 ° C overnight. NaBH ₃ CN (4.08 g, 64.8 mmol, 1.2 equivalents) was then added to the mixture. After stirring at room temperature for 5 h, the mixture was filtered. The filtrate was concentrated to obtain a yellow oil. Chlorosulfonic acid (30 mL) was cooled to -10 ° C, and the above-mentioned crude product was added dropwise. The reaction mixture was heated to 100 ° C for 10 min, then cooled and poured into ice. The mixture was neutralized with 2M NaOH and extracted with ethyl acetate. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=2/1, v/v) to give 6-bromo-3-methyl-isoquinoline (4.0 g, 34% yield over 3 steps) as a yellow solid.

Step 2: Preparation of methyl 3-methylisoquinoline-6-carboxylate

To an autoclaved vessel was added 6-bromo-3-methyl-isoquinoline (4.0 g, 18 mmol), Pd(dppf) _Cl2 (735 mg, 0.9 mmol, 0.05 eq) and triethylamine (5.0 mL, 36 mmol, 2 eq) in 40 mL of methanol. The vessel was purged three times with nitrogen and three times with carbon monoxide. The vessel was pressurized to 3 MPa with carbon monoxide and heated to 100°C. The reaction was stirred overnight and then allowed to cool to room temperature. The resulting solution was concentrated and purified by flash chromatography on silica gel (PE/EtOAc = 1/1, v/v) to give methyl 3-methylisoquinoline-6-carboxylate (3.4 g, 94%) as a white solid.

Step 3: Preparation of (3-methyl-isoquinolin-6-yl)-methanol

To a solution of methyl 3-methylisoquinoline-6-carboxylate (3.3 g, 16.42 mmol, 1 eq) in anhydrous THF (100 mL) was added LiAlH(t-BuO) ₃ (12.5 g, 45.25 mmol, 3 eq). The resulting mixture was stirred at 60 ° C for 5 h and then quenched by adding water. The mixture was extracted with ethyl acetate. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=1/1, v/v) to give (3-methyl-isoquinolin-6-yl)-methanol (2.5 g, 89%) as a white solid.

Step 4: Preparation of 2-((3-methylisoquinolin-6-yl)methyl)isonicotinic acid

The title compound was synthesized as described in Intermediate 3, Steps 4, 5 and 6.

Intermediates 8-14

The title compound was synthesized as described for Intermediate 7.

Intermediate 15: Preparation of 5-(aminomethyl)-4,6-dimethylpyridin-2-amine hydrochloride

Step 1: Preparation of 5-iodo-4,6-dimethyl-pyridin-2-ylamine

A mixture of 4,6-dimethyl-pyridin-2-ylamine (6 g, 49.1 mmol, 1.0 equiv), periodic acid (1.6 g, 7.37 mmol, 0.15 equiv) and iodine (6.2 g, 24.5 mmol, 0.5 equiv) was added to a mixed solution of acetic acid (120 mL), H ₂ O ₂ (6 mL) and H ₂ SO ₄ (1 mL) at 80° C. for 4 h, then the reaction mixture was poured into a 10% aqueous Na ₂ S ₂ O ₃ solution to quench any unrecalled iodine and extracted with ether. The extract was washed with a 10% aqueous NaOH solution, dried over Na ₂ SO ₄ and concentrated, and the resulting residue was purified by silica gel chromatography (DCM / MeOH = 100 / 1, v / v) to give 5-iodo-4,6-dimethyl-pyridin-2-ylamine (10 g, 80%) as a yellow solid.

Step 2: Preparation of 6-amino-2,4-dimethyl-nicotinonitrile

To a solution of 5-iodo-4,6-dimethyl-pyridin-2-ylamine (10 g, 40.3 mmol, 1.0 equiv) in DMF (300 mL) was carefully added Zn(CN) ₍ 14 g, 120.9 mmol, 3.0 equiv) _and Pd( _PPh ) (4.65 g, 4.03 mmol, 0.1 equiv). The mixture was stirred at 90 ° C. overnight under N _2. Ethyl acetate and water were added. The organic layer was separated and concentrated. The resulting residue was purified by silica gel chromatography (DCM/MeOH=100/1, v/v) to give 6-amino-2,4-dimethyl-nicotinonitrile (5 g, 84%) as a yellow solid.

Step 3: Preparation of tert-butyl (6-amino-2,4-dimethylpyridin-3-yl)methylcarbamate

To a solution of 6-amino-2,4-dimethyl-nicotinonitrile (8.1 g, 55 mmol, 1.0 equivalent) in THF (300 mL) was slowly added BH ₃ .MeS ₂ (10 M, 55 mL, 550 mmol, 10.0 equivalent) at room temperature. The mixture was stirred at reflux for 48 h. After cooling to room temperature, the mixture was quenched by the addition of concentrated HCl. The mixture was basified to pH 8 with saturated NaHCO ₃ solution. TEA (9.2 mL, 66 mmol, 1.2 equivalents) and Boc ₂ O (14.4 g, 66 mmol, 1.2 equivalents) were added to the mixture. The reaction mixture was stirred at room temperature for 1 h and then extracted with ethyl acetate. The combined organic layers were dried over Na ₂ SO ₄ , filtered, and concentrated. The residue was purified on a silica gel column (PE/EtOAc=1/1) to give tert-butyl (6-amino-2,4-dimethylpyridin-3-yl)methylcarbamate (4.1 g, 30%) as a yellow solid.

Step 4: Preparation of 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine hydrochloride

To a solution of tert-butyl (6-amino-2,4-dimethylpyridin-3-yl)methylcarbamate (4.1 g, 16.3 mmol, 1.0 equiv) in ethyl acetate (20 mL) was added a solution of HCl in ethyl acetate (10 M, 50 mL). The mixture was stirred at room temperature for 1 h, and the precipitate was collected by filtration to give 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine hydrochloride (2.0 g, 66%) as a white solid.

Intermediate 16: Preparation of 6-(aminomethyl)isoquinolin-1-amine

Step 1: Preparation of 6-bromo-1-chloroisoquinoline

To a solution of 6-bromoisoquinolin-1-ol (10 g, 44.4 mmol, 1.0 equiv) in POCl ₃ (200 mL) was added. The mixture was stirred at 120° C. overnight. The mixture was concentrated and extracted with DCM, and the combined extracts were washed with brine, dried, and concentrated to give 6-bromo-1-chloroisoquinoline (10 g, crude) as a yellow solid.

Step 2: Preparation of 6-bromoisoquinolin-1-amine

A solution of 6-bromo-1-chloroisoquinoline (10 g, 41.50 mmol, 1.0 equiv) in MeOH/NH ₃ (500 mL) was stirred for 48 h at 30° C. The mixture was concentrated to give 6-bromoisoquinolin-1-amine (10 g, crude) as a yellow solid.

Step 3: Preparation of 1-aminoisoquinoline-6-carbonitrile

To a solution of 6-bromoisoquinolin-1-amine (10 g, 45.04 mmol, 1.0 eq) in DMF (400 mL) was added Zn(CN) (13.22 _g , 112.3 mmol, 2.5 eq) and Pd( _pph ) ₍ 5.2 g, 4.51 mmol, 0.1 eq). The mixture was stirred at 120° C. for 4 h. The mixture was concentrated and extracted with DCM. The combined extracts were dried and concentrated. The residue was purified by flash chromatography on a silica gel column (DCM/MeOH=50/1, v/v) to give 1-aminoisoquinoline-6-carbonitrile (4.5 g, 59%) as a yellow oil.

Step 4: Preparation of 6-(aminomethyl)isoquinolin-1-amine

To a solution of 1-aminoisoquinoline-6-carbonitrile (4.5 g, 26.47 mmol, 1 eq) in MeOH (200 mL), DMF (200 mL) and ammonium hydroxide (100 mL) was added Raney nickel (4.0 g). The mixture was stirred at 40 ° C. overnight under a hydrogen atmosphere. The Raney nickel was filtered off, and the filtrate was concentrated in vacuo to give 6-(aminomethyl)isoquinolin-1-amine (3 g, crude) as a yellow solid.

Intermediate 17: Preparation of (6-fluoro-1H-indol-5-yl)methanamine

Step 1: Preparation of 6-fluoro-1-(triisopropylsilyl)-1H-indole-5-carbaldehyde

At -78 ° C, s-BuLi (6.3 mL, 1.3 M, 1.2 eq) was slowly added to a solution of 6-fluoro-1-(triisopropylsilyl)-1H-indole (2 g, 6.9 mmol, 1.0 eq) in THF (30 mL) . The mixture was then stirred at this temperature for 1 h. DMF (1.5 g, 20.7 mmol, 3.0 eq) was added dropwise. The mixture was stirred at -78 ° C for 1 h. The reaction was then quenched with saturated NH ₄ Cl aqueous solution. The obtained mixture was extracted with EtOAc (50 mL × 3). The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified on a silica gel column (PE / EtOAc = 100 / 1) to give 6-fluoro-1-(triisopropylsilyl)-1H-indole-5-carbaldehyde (950 mg, 57%) as a yellow oil.

Step 2: Preparation of 6-fluoro-1H-indole-5-carbaldehyde oxime

A mixture of 6-fluoro-1-(triisopropylsilyl)-1H-indole-5-carbaldehyde (780 mg, 2.45 mmol, 1.0 eq) and _NH OH.HCl (340 mg, 4.89 mmol, 2.0 eq) in NH ₃ /MeOH (15% w/w, 10 mL) was stirred at room temperature overnight. The mixture was concentrated. The residue was directly purified on a silica gel column (PE/EtOAc=50/1) to give 6-fluoro-1H-indole-5-carbaldehyde oxime (460 mg, crude) as a yellow solid.

Step 3: Preparation of (6-fluoro-1H-indol-5-yl)methanamine

A mixture of 6-fluoro-1H-indole-5-carbaldehyde oxime (460 mg, 1.38 mmol, 1.0 equiv) and Raney nickel (100 mg) in NH ₃ /MeOH (15% w / w, 10 mL) was stirred at room temperature overnight under an atmosphere of H ₂ (1 atm). The mixture was filtered and concentrated to give (6-fluoro-1H-indol-5-yl)methanamine (420 mg, 95%) as a gray solid. The solid was used in the next step without further purification.

Intermediate 18: Preparation of (5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)methanamine

Step 1: Preparation of 5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde

To a suspension of 5-chloro-1H-pyrrolo[2,3-b]pyridine (10.0 g, 65.5 mmol, 1 equivalent) in AcOH (56.7 mL) and water (28.3 mL) was added hexamethylenetetramine (11.9 g, 85.2 mmol, 1.3 equivalents). The mixture was stirred under reflux overnight, followed by the addition of 200 mL of water. After stirring for 30 min, the reaction mixture was filtered to recover the solid, which was then dried in air to give 5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carboxaldehyde (6.2 g, 53%) as a yellow solid.

Step 2: Preparation of 5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde oxime

To a solution of 5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde (1.0 g, 5.55 mmol, 1 eq) in EtOH (40 mL) and water (10 mL) was added hydroxylammonium chloride (575 mg, 8.33 mmol, 1.5 eq) and Na ₂ CO ₃ (1.06 g, 10.0 mmol, 1.8 eq). The mixture was stirred at reflux for 3 h and then cooled to room temperature. The precipitate was collected by filtration to give 5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde oxime (1.0 g, 92%) as an off-white solid.

Step 3: Preparation of (5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)methanamine

To a solution of 5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbaldehyde oxime (200 mg, 2 mmol, 1 eq) in MeOH (5 mL) was added _NiCl₂ (128 mg, 1 mmol, 1 eq) and _NaBH₄ (228 mg, 6 mmol, 6 eq). The mixture was stirred at room temperature under a hydrogen atmosphere for 5 h. _NiCl₂ was filtered off, and the filtrate was concentrated in vacuo to afford (5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)methanamine (65 mg, 30%) as a yellow solid.

Intermediate 19: Preparation of (5-chloro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-3-yl)methanamine

Step 1: Preparation of 1-(2-amino-5-chlorophenyl)-2-chloroethanone

To a stirred solution of boron trichloride in toluene (200 mL, 1 M, 0.2 mol, 1.16 equiv) was added dropwise a solution of 4-chloroaniline (22.0 g, 0.172 mol, 1.0 equiv) in anhydrous toluene (200 mL) under nitrogen at a temperature ranging from 5°C to 10°C. Chloroacetonitrile (15 mL, 0.237 mol, 1.38 equiv) and aluminum chloride (29.0 g, 0.217 mol, 1.26 equiv) were added sequentially to the resulting mixture. The mixture was refluxed for 18 h. After cooling, ice-cold hydrochloric acid (2N, 500 mL) was added, and a yellow precipitate formed. The mixture was warmed at 80°C with stirring until the precipitate dissolved. The cooled solution was extracted with dichloromethane ₍ 250 mL x 3). The organic layer was washed with water, dried ( _Na₂SO₄ ), and concentrated. The obtained residue was purified on a silica gel column (PE/EtOAc=50/1 to PE/EtOAc/DCM=1/8/1, v/v/v) to give 1-(2-amino-5-chlorophenyl)-2-chloroethanone (18.3 g, 52%) as a yellow-brown solid.

Step 2: Preparation of 5-chloro-3-(chloromethyl)-1H-indazole

To a stirred suspension of 1-(2-amino-5-chlorophenyl)-2-chloroethanone (16 g, 78 mmol, 1.0 eq) in concentrated hydrochloric acid (120 mL) at 0°C was added a solution of sodium nitrite (5.9 g, 86 mmol, 1.1 eq) in water (30 mL). After 1 h, a solution of _SnCl2.2H2O (42.3 _g , 187 mmol, 2.4 eq) in concentrated hydrochloric acid (60 mL) was added to the reaction mixture and stirred for 1 h. Ice-water was added to the reaction mixture. The precipitate was collected by filtration, washed with water, and dried to give crude 5-chloro-3-(chloromethyl)-1H-indazole, which was used in the next step without further purification (13.5 g, 86%).

Step 3: Preparation of 5-chloro-3-(chloromethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole

A solution of 5-chloro-3-(chloromethyl)-1H-indazole (13.5 g, 67 mmol, 1.0 equiv), 3,4-dihydro-2H-pyran (11.3 g, 134 mmol, 2.0 equiv) and p-toluenesulfonic acid monohydrate (1.27 g, 6.7 mmol, 0.1 equiv) in THF (300 mL) was stirred at 70 ° C for 12 h. After cooling to room temperature (about 22 ° C), the reaction mixture was mixed with water (300 mL) and extracted with ethyl acetate (200 mL × 2). The organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ , and concentrated to give 5-chloro-3-(chloromethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (16 g, 84%) as a yellow solid.

Step 4: Preparation of 2-((5-chloro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-3-yl)methyl)isoindoline-1,3-dione

To a solution of 5-chloro-3-(chloromethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (18 g, 63 mmol, 1 equiv) in anhydrous DMF (200 mL) under N ₂ was added potassium phthalimide (17.5 g, 94 mmol, 1.5 equiv), and the resulting mixture was heated at 90 ° C for 2 h. The mixture was poured into water and extracted with DCM (200 mL×2). The combined organic layers were washed with water and brine, dried over anhydrous Na ₂ SO ₄ and concentrated. The crude product was washed with EtOH to give 2-((5-chloro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-3-yl)methyl)isoindoline-1,3-dione (15 g, 60%) as a white solid.

Step 5: Preparation of (5-chloro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-3-yl)methanamine

To a solution of 2-((5-chloro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-3-yl)methyl)isoindoline-1,3-dione (15 g, 37.9 mmol, 1.0 equiv) in THF (300 mL) and DCM (60 mL) was added hydrazine hydrate (9.5 g, 189 mmol, 5 equiv). The white suspension was stirred at 48 ° C for 12 h, and phthalic acid hydrazide was removed by filtration. The filtrate was concentrated in vacuo, and the crude material was dissolved in DCM and washed with 1N NaOH solution. The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated to give (5-chloro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-3-yl)methanamine (9.9 g, 99%) as a yellow solid.

Intermediate 20: Preparation of 5-(aminomethyl)-6-methylpyridin-2-amine

A solution of 6-amino-2-methylnicotinonitrile (200 mg, 1.5 mmol, 1.0 equiv) and Raney nickel (50 mg) in MeOH (10 mL) was stirred at room temperature overnight under H ₂ (1 atm). The mixture was then filtered and the filtrate was concentrated to give 5-(aminomethyl)-6-methylpyridin-2-amine (210 mg, quantitative) as a yellow solid. The solid was used without further purification.

Intermediate 21: Preparation of 5-(aminomethyl)-4-methylpyridin-2-amine

A solution of 6-amino-4-methylnicotinonitrile (200 mg, 1.5 mmol, 1.0 equiv) and Raney nickel (50 mg) in MeOH (10 mL) was stirred at room temperature overnight under H ₂ (1 atm). The mixture was then filtered and the filtrate was concentrated to give 5-(aminomethyl)-4-methylpyridin-2-amine (210 mg, quantitative) as a yellow solid. The solid was used without further purification.

Intermediate 22: Preparation of 5-(aminomethyl)-6-(trifluoromethyl)pyridin-2-amine

A solution of 6-amino-2-(trifluoromethyl)nicotinonitrile (200 mg, 1.06 mmol, 1.0 equiv) and Raney nickel (50 mg) in MeOH (10 mL) was stirred at room temperature overnight under H ₂ (1 atm). The mixture was then filtered and the filtrate was concentrated to give 5-(aminomethyl)-6-(trifluoromethyl)pyridin-2-amine (204 mg, quantitative) as a yellow solid. The solid was used without further purification.

Intermediate 23: Preparation of tert-butyl 5-(aminomethyl)benzo[d]isoxazol-3-ylcarbamate

Step 1: Preparation of 5-methylbenzo[d]isoxazol-3-amine

Potassium tert-butoxide (4.57 g, 40.8 mmol, 1.1 equivalents) is suspended in THF (40 mL). Acetone oxime (2.97 g, 40.7 mmol, 1.1 equivalents) is added, and the mixture is stirred at room temperature for 20 min, followed by dropwise addition of a THF (30 mL) solution of 2-fluoro-5-methylbenzonitrile (5.00 g, 37 mmol, 1.0 equivalents). The mixture is stirred at room temperature for 3 h, then refluxed overnight. The dark brown solution is quenched with water (10 mL). The mixture is distributed between a saturated NaHCO ₃ aqueous solution (50 mL) and ethyl acetate (150 mL). The aqueous layer is extracted with ethyl acetate (50 mL). The combined organic layers are dried over anhydrous Na ₂ SO ₄ and concentrated to obtain a brown oil. The crude oil is dissolved in EtOH (80 mL). H ₂ O (53 mL) and concentrated HCl (27 mL) are added, and the mixture is stirred at 90 ° C for 2 h. The mixture was cooled to room temperature and basified to pH 10 with aqueous NaOH. The aqueous layer was extracted with ethyl acetate (100 mL x 3). The combined organic layers were dried over _{anhydrous Na2SO4} and concentrated. The residue was purified by flash chromatography (PE/EtOAc = 5/1, v/v) to give 5-methylbenzo[d]isoxazol-3-amine (2.5 g, 45.6%) as a white solid.

Step 2: Preparation of 5-methylbenzo[d]isoxazol-3-amine bis(tert-butyl carbamate)

A mixture of 5-methylbenzo[d]isoxazol-3-amine (1.48 g, 10 mmol, 1.0 equiv), _Boc2O (6.54 g, 30 mmol, 3.0 equiv), DMAP (122 mg, 1.0 mmol, 0.1 equiv), and TEA (4.2 mL, 30 mmol, 3.0 equiv) in DCM (30 mL) was refluxed for 18 h. The mixture was washed with water (30 mL x 2), dried over _{anhydrous Na2SO4} , and concentrated. The residue was purified by flash chromatography (PE/EtOAc/DCM = 1/20/1 to 1/7/1, v/v/v) to give 5-methylbenzo[d]isoxazol-3-amine bis(tert-butylcarbamate) (3.2 g, 92%) as a white solid.

Step 3: Preparation of 5-(bromomethyl)benzo[d]isoxazol-3-amine bis(tert-butyl carbamate)

A mixture of 5-methylbenzo[d]isoxazol-3-amine bis(tert-butyl carbamate) (1.04 g, 3 mmol, 1.0 equiv), NBS (536 mg, 3 mmol, 1.0 equiv), and AIBN (53 mg, 0.32 mmol, 0.1 equiv) in CCl ( ₃₀ mL) was stirred at 85° C. for 5 h. The mixture was cooled to room temperature and filtered. The filtrate was concentrated and the residue was purified by flash chromatography (PE/EtOAc=10/1, v/v) to afford 5-(bromomethyl)benzo[d]isoxazol-3-amine bis(tert-butyl carbamate) (970 mg, 75.8%) as a white solid.

Step 4: Preparation of 2-((3-aminobenzo[d]isoxazol-5-yl)methyl)isoindoline-1,3-dione bis(tert-butylcarbamate)

A mixture of 5-(bromomethyl)benzo[d]isoxazol-3-amine bis(tert-butylcarbamate) (602 mg, 1.4 mmol, 1.0 equiv), isoindoline-1,3-dione (310 mg, 2.1 mmol, 1.5 equiv), _{and Cs2CO3} (1.1 g, 3.4 mmol, 2.4 equiv) in DMF (10 mL) was stirred at 20°C overnight. The mixture was partitioned between ethyl acetate (20 mL) and water (20 mL). The aqueous layer was extracted with ethyl acetate (20 mL x 2). The combined organic layers were dried over _{anhydrous Na2SO4} and concentrated. The residue was purified by flash chromatography (PE/EtOAc=5/1, v/v) to give 2-((3-aminobenzo[d]isoxazol-5-yl)methyl)isoindoline-1,3-dione bis(tert-butyl carbamate) (616 mg, 88.8%) as a white solid.

Step 5: Preparation of tert-butyl 5-(aminomethyl)benzo[d]isoxazol-3-ylcarbamate

To a solution of 2-((3-aminobenzo[d]isoxazol-5-yl)methyl)isoindoline-1,3-dione bis(tert-butyl carbamate) (320 mg, 0.65 mmol, 1.0 equiv) in n-BuOH (10 mL) was added _NH NH _{.H 0} (0.20 mL). The mixture was stirred at room temperature overnight. The white slurry was diluted with DCM (10 mL) and filtered. The filtrate was evaporated and the residue was triturated with Et ₀ and dried to give tert-butyl 5-(aminomethyl)benzo[d]isoxazol-3-ylcarbamate (120 mg, 70.6%) as a white solid.

Intermediate 24: Preparation of 2-((6-methyl-2-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinic acid

Step 1: Preparation of 2-methanesulfonyl-6-methyl-quinoline-3-carbaldehyde

To a suspension of 2-chloro-6-methyl-quinoline-3-carboxaldehyde (1.0 g, 4.88 mmol, 1 equivalent) in DMF (30 mL) was added sodium methanesulfonate (1.49 g, 14.6 mmol, 3 equivalents). The reaction mixture was stirred at 100 ° C for 1 h under nitrogen. The cooled mixture was partitioned between EtOAc and water. The organic layer was separated and concentrated, and the resulting residue was purified by chromatography on a silica gel column (EtOAc/PE=1/2, v/v) to give 2-methanesulfonyl-6-methyl-quinoline-3-carboxaldehyde (1.1 g, 90%) as a yellow solid.

Step 2: Preparation of (2-methanesulfonyl-6-methyl-quinolin-3-yl)-methanol

To a solution of 2-methanesulfonyl-6-methyl-quinoline-3-carbaldehyde (400 mg, 1.61 mmol, 1 equiv) in MeOH (20 mL) was added NaBH ₄ (67 mg, 1.77 mmol, 1.1 equiv). The reaction mixture was stirred at 0° C. for 1 h. The mixture was partitioned between DCM and water. The organic layer was separated and concentrated. The resulting residue was purified by chromatography on a silica gel column (EtOAc/PE=1/2, v/v) to give (2-methanesulfonyl-6-methyl-quinoline-3-yl)-methanol (395 mg, 97%) as a yellow solid.

Step 3: Preparation of 3-chloromethyl-2-methanesulfonyl-6-methyl-quinoline

A mixture of (2-methanesulfonyl-6-methyl-quinolin-3-yl)-methanol (395 mg, 1.57 mmol, 1.0 equiv) in SOCl ₂ (10 mL) was stirred at room temperature for 1 h and concentrated. The resulting residue was dissolved in DCM and treated with saturated NaHCO ₃ solution. The organic layer was concentrated to give 3-chloromethyl-2-methanesulfonyl-6-methyl-quinoline (400 mg, 95%) as a yellow solid.

Step 4: Preparation of methyl 2-((6-methyl-2-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinate

To a solution of 3-chloromethyl-2-methanesulfonyl-6-methyl-quinoline (400 mg, 1.49 mmol, 1.0 equiv) in dioxane (15 mL) was added methyl 2-(trimethylstannyl)isonicotinate (492 mg, 1.64 mmol, 1.1 equiv) and Pd(PPh ₃ ) ₂ Cl ₂ (105 mg, 0.15 mmol, 0.1 equiv). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, the solvent was removed, and the mixture was finally purified by chromatography on a silica gel column (DCM/MeOH=100/1, v/v) to give methyl 2-((6-methyl-2-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinate (100 mg, 18%) as a yellow solid.

Step 5: Preparation of 2-((6-methyl-2-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinic acid

To a solution of 2-((6-methyl-2-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinate (100 mg, 0.27 mmol, 1 eq) in THF/H ₂ O (5 mL, 1:1) was added LiOH.H ₂ O (17 mg, 0.41 mmol, 1.5 eq). The mixture was stirred at room temperature for 1 h and acidified to pH 3 with 1 N HCl solution. The solvent was removed to give 2-((6-methyl-2-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinate as a yellow solid, which was used in the next step without further purification.

Intermediate 25: Preparation of 2-((3-cyanoquinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of (3-bromo-quinolin-6-yl)-methanol

To a solution of 3-bromoquinoline-6-formic acid methyl ester (4.5 g, 17.0 mmol, 1 equivalent) in THF (200 mL) was added LiAlH (t-BuO) ₃ (10.78 g, 42.5 mmol, 2.5 equivalents). The resulting mixture was stirred at 40 ° C for 12 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE / EtOAc = 2 / 1, v / v) to give (3-bromo-quinoline-6-yl) -methanol (3.1 g, 78%) as a yellow solid.

Step 2: Preparation of 6-hydroxymethyl-quinoline-3-carbonitrile

To a solution of (3-bromo-quinolin-6-yl)-methanol (3.1 g, 13.1 mmol, 1 eq) in DMF (60 mL) was added Zn(CN) ₍ 1.52 g, 13.1 mmol, 1 eq) and Pd( _PPh ) ₍ 757 mg, 0.66 mmol, 0.05 eq). The mixture was stirred at 90 ° C overnight and then cooled to room temperature. The precipitate was filtered off and the filtrate was concentrated. The residue was purified by flash chromatography on a silica gel column (PE/EtOAc=2/1, v/v) to give 6-hydroxymethyl-quinoline-3-carbonitrile (1.95 g, 81%) as a yellow solid.

Step 3: Preparation of 6-chloromethyl-quinoline-3-carbonitrile

A mixture of 6-hydroxymethyl-quinoline-3-carbonitrile (1.95 g, 10.6 mmol, 1.0 equiv) in SOCl ₂ (50 mL) was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DCM and treated with saturated NaHCO ₃ solution. The organic layer was concentrated to give 6-chloromethyl-quinoline-3-carbonitrile (2.0 g, 93%) as a yellow solid.

Step 4: Preparation of methyl 2-((3-cyanoquinolin-6-yl)methyl)isonicotinate

To a solution of 6-chloromethyl-quinoline-3-carbonitrile (2.0 g, 10.9 mmol, 1 eq) in dioxane (60 mL) was added methyl 2-(trimethylstannyl)isonicotinate (3.60 g, 12.0 mmol, 1.1 eq) and Pd(PPh ₃ ) ₂ Cl ₂ (770 mg, 0.11 mmol, 0.1 eq). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, the solvent was removed, and the product was purified by chromatography on a silica gel column (DCM/MeOH=100/1, v/v) to give methyl 2-((3-cyanoquinolin-6-yl)methyl)isonicotinate (750 mg, 23%) as a yellow solid.

Step 5: Preparation of 2-((3-cyanoquinolin-6-yl)methyl)isonicotinic acid

To a solution of methyl 2-((3-cyanoquinolin-6-yl)methyl)isonicotinate (750 mg, 2.47 mmol, 1 eq) in THF/H ₂ O (20 mL, 1:1) was added LiOH.H ₂ O (156 mg, 3.71 mmol, 1.5 eq). The mixture was stirred at room temperature for 1 h and then acidified to pH 3 with 1 N HCl solution. The solvent was concentrated to give 2-((3-cyanoquinolin-6-yl)methyl)isonicotinoic acid as a yellow solid, which was used in the next step without further purification.

Intermediate 26: Preparation of 2-((3-chloro-8-(methoxycarbonyl)quinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of methyl 3-chloro-8-iodoquinoline-6-carboxylate

To a solution of 8-iodoquinoline-6-carboxylic acid methyl ester (30 g, 96 mmol, 1.0 equiv) in AcOH (1.0 L) was added NCS (38 g, 293 mmol, 3 equiv). The mixture was stirred at 100 ° C overnight. The mixture was concentrated in vacuo, and the residue was purified by chromatography on a silica gel column (PE / DCM = 1 / 1, v / v) to give 3-chloro-8-iodoquinoline-6-carboxylic acid methyl ester (15 g, 49%) as a yellow solid.

Step 2: Preparation of (3-chloro-8-iodo-quinolin-6-yl)-methanol

To a solution of 3-chloro-8-iodoquinoline-6-methyl formate (12 g, 34.5 mmol, 1.0 equivalent) in anhydrous THF (200 mL) was carefully added lithium tri-tert-butoxyaluminum hydride (22 g, 70 mmol, 3.4 equivalents). The mixture was stirred at 50 ° C for 5 h under _N protection. EtOAc and water were then added. The organic layer was concentrated and purified by chromatography on a silica gel column (PE/DCM=1/1, v/v) to obtain (3-chloro-8-iodo-quinoline-6-yl)-methanol (7.6 g, 69%) as a white solid.

Step 3: Preparation of 3-chloro-6-hydroxymethyl-quinoline-8-carbonitrile

To a solution of (3-chloro-8-iodo-quinolin-6-yl)-methanol (7.6 g, 23.8 mmol, 1.0 equiv) in DMF (100 mL) was carefully added Zn(CN) ( _2.79 g, 23.8 mmol, 1.0 _equiv ) and Pd(PPh) ₍ 2.75 g, 2.38 mmol, 0.1 equiv). The mixture was stirred at 50 ° C. overnight under _N protection. EtOAc and water were then added. The organic layer was concentrated and purified by chromatography on a silica gel column (PE/DCM=1/2, v/v) to give 3-chloro-6-hydroxymethyl-quinoline-8-carbonitrile (5.0 g, 96%) as a yellow solid.

Step 4: Preparation of 3-chloro-6-chloromethyl-quinoline-8-carbonitrile

A mixture of 3-chloro-6-hydroxymethyl-quinoline-8-carbonitrile (2.9 g, 13.3 mmol, 1.0 equiv) in _SOCl2 (50 mL) was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DCM and treated with saturated _NaHCO3 solution to give 3-chloro-6-chloromethyl-quinoline-8-carbonitrile (2.2 g, 70%) as a yellow solid.

Step 5: Preparation of methyl 2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinate

To a solution of 3-chloro-6-chloromethyl-quinoline-8-carbonitrile (2.0 g, 8.47 mmol, 1.0 equiv) in dioxane (40 mL) was added methyl 2-(trimethylstannyl)isonicotinate (2.8 g, 9.32 mmol, 1.1 equiv) and Pd(PPh ₃ ) ₂ Cl ₂ (597 mg, 0.85 mmol, 0.1 equiv). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, concentrated, and purified by chromatography on a silica gel column (DCM/MeOH=100/1, v/v) to give methyl 2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinate (1.4 g, 49%) as a yellow solid.

Step 6: Preparation of methyl 3-chloro-6-((4-(methoxycarbonyl)pyridin-2-yl)methyl)quinoline-8-carboxylate

A mixture of methyl 2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinate (1.20 g, 3.56 mmol, 1 equiv) in HCl/MeOH (10 N, 100 mL) was heated at 80° C. for 7 days and then concentrated. The residue was dissolved in DCM and treated with saturated NaHCO ₃ solution. The organic layer was concentrated, and the residue was purified by flash chromatography on a silica gel column (PE/EtOAc=1/1, v/v) to give methyl 3-chloro-6-((4-(methoxycarbonyl)pyridin-2-yl)methyl)quinoline-8-carboxylate (900 mg, 68%) as a yellow solid.

Step 7: Preparation of 2-((3-chloro-8-(methoxycarbonyl)quinolin-6-yl)methyl)isonicotinic acid

To a solution of methyl 3-chloro-6-((4-(methoxycarbonyl)pyridin-2-yl)methyl)quinoline-8-carboxylate (900 mg, 2.43 mmol, 1 eq) in THF/H ₂ O (20 mL, 1:1) was added LiOH.H ₂ O (102 mg, 2.43 mmol, 1.0 eq). The mixture was stirred at room temperature for 1 h and acidified to pH 3 with 1N HCl solution. The mixture was extracted with DCM, and the combined organic layers were dried and concentrated. The residue was purified by flash chromatography on a silica gel column (DCM/MeOH=10/1, v/v) to give 2-((3-chloro-8-(methoxycarbonyl)quinolin-6-yl)methyl)isonicotinic acid (400 mg, 46%) as a yellow solid.

Intermediate 27: Preparation of 2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of methyl 4-amino-3-iodobenzoate

To a solution of 4-aminobenzoic acid methyl ester (20g, 0.132mol, 1 equivalent) in AcOH (500mL) was added a solution of ICl (23.6g, 0.146mol, 1.1 equivalents) in AcOH (500mL) at 0°C. The mixture was stirred at room temperature for 2h. AcOH was concentrated under reduced pressure. The residue was diluted with DCM and washed with saturated NaHCO _3. The aqueous layer was extracted with DCM, and the combined extracts were dried and concentrated. The resulting residue was purified by chromatography on a silica gel column (EtOAc/PE=1/15, v/v) to obtain 4-amino-3-iodobenzoic acid methyl ester (27.4g, 75%) as an off-white solid.

Step 2: Preparation of methyl 8-iodo-3-methylquinoline-6-carboxylate

The mixture of 4-amino-3-iodobenzoic acid methyl esters (26g, 93.5mmol), 2-methyl-acrolein (24.5g, 0.28mol, ₃ equivalents) and 6N HCl (95mL) is heated to reflux 24h.Then the mixture is cooled and adjusted to pH～5-6 with saturated NaHCO.The mixture is extracted with DCM.The organic layer merged is washed with salt water, dried over _MgSO , filtered, then concentrated, and purified by chromatography on silica gel column (EtOAc/PE=1/20, v/v), to obtain 8-iodo-3-methylquinoline-6-methyl-formiate (10.2g, 33%) as a yellow solid.

Step 3: Preparation of (8-iodo-3-methyl-quinolin-6-yl)-methanol

To a THF (200 mL) solution of 8-iodo-3-methylquinoline-6-formic acid methyl ester (7.5 g, 22.9 mmol, 1 equivalent) was added LiAlH (t-BuO) ₃ (14.6 g, 57.3 mmol, 2.5 equivalents). The resulting mixture was stirred at 40 ° C for 12 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE / EtOAc = 2 / 1, v / v) to give (8-iodo-3-methyl-quinoline-6-yl) -methanol (6.5 g, 95%) as a yellow solid.

Step 4: Preparation of 6-hydroxymethyl-3-methyl-quinoline-8-carbonitrile

To a solution of (8-iodo-3-methyl-quinolin-6-yl)-methanol (2.1 g, 7.0 mmol, 1 eq) in DMF (50 mL) was added Zn(CN) ₍ 815 mg, 7.0 mmol, ₁ eq) and Pd( _PPh ) (404 mg, 0.35 mmol, 0.05 eq). The mixture was stirred at 90 ° C overnight and then cooled to room temperature. The precipitate was filtered off and the filtrate was concentrated. The resulting residue was purified by flash chromatography on a silica gel column (PE/EtOAc=2/1, v/v) to give 6-hydroxymethyl-3-methyl-quinoline-8-carbonitrile (1.2 g, 86%) as a yellow solid.

Step 5: Preparation of 6-chloromethyl-quinoline-3-carbonitrile

A mixture of 6-hydroxymethyl-3-methyl-quinoline-8-carbonitrile (1.2 g, 6.06 mmol, 1.0 equiv) in SOCl ₂ (30 mL) was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DCM and treated with saturated NaHCO ₃ solution. The organic layer was concentrated to give 6-chloromethyl-quinoline-3-carbonitrile (1.2 g, 92%) as a yellow solid.

Step 6: Preparation of methyl 2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinate

To a solution of 6-chloromethyl-quinoline-3-carbonitrile (1.2 g, 5.55 mmol, 1 eq) in dioxane (60 mL) was added methyl 2-(trimethylstannyl)isonicotinate (1.84 g, 6.11 mmol, 1.1 eq) and Pd(PPh ₃ ) ₂ Cl ₂ (428 mg, 0.61 mmol, 0.1 eq). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, the solvent was removed, and the product was purified by chromatography on a silica gel column (DCM/MeOH=100/1, v/v) to give methyl 2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinate (1.1 g, 62%) as a yellow solid.

Step 7: Preparation of 2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinic acid

To a solution of methyl 2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinate (1.1 g, 3.47 mmol, 1 eq) in THF/H ₂ O (30 mL, 1:1) was added LiOH.H ₂ O (219 mg, 5.21 mmol, 1.5 eq). The mixture was stirred at room temperature for 1 h and acidified to pH 3 with 1N HCl solution. The mixture was extracted with DCM, and the combined organic layers were dried and concentrated to give 2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinoic acid (950 mg, 91%) as a white solid, which was used without further purification.

Intermediate 28: Preparation of 2-((3-methyl-1H-indol-5-yl)methyl)isonicotinic acid

Step 1: Preparation of methyl 3-chloro-1H-indole-5-carboxylate

To a MeOH solution of 1H-indole-5-methyl-formiate (10.0 g, 57.1 mmol, 1.0 equivalent) was added NCS (8.4 g, 62.8 mmol, 1.1 equivalents). The mixture was stirred at room temperature for 3 h. MeOH was removed and the resulting residue was dissolved in EtOAc. The mixture was washed twice with brine. The organic layer was dried and concentrated to give 3-chloro-1H-indole-5-methyl-formiate (quantitative) as a yellow solid.

Step 2: Preparation of 1-tert-butyl 5-methyl 3-chloro-1H-indole-1,5-dicarboxylate

To a solution of methyl 3-chloro-1H-indole-5-carboxylate (11.9 g, 57.1 mmol, 1.0 equiv) in MeOH was added Boc ₂ O (18.7 g, 86.7 mmol, 1.5 equiv) and DMAP (348 mg, 2.86 mmol, 0.05 equiv). The mixture was stirred at room temperature for 2 h. The mixture was concentrated and purified by chromatography on a silica gel column (EtOAc/PE=1/10, v/v) to give 1-tert-butyl 5-methyl 3-chloro-1H-indole-1,5-dicarboxylate (13.4 g, 76%) as an off-white solid.

Step 3: Preparation of tert-butyl 3-chloro-5-(hydroxymethyl)-1H-indole-1-carboxylate

To a solution of 1-tert-butyl 5-methyl 3-chloro-1H-indole-1,5-dicarboxylate (7.0 g, 22.6 mmol, 1 equivalent) in THF (100 mL) was added LiAlH(t-BuO) ₃ (14.4 g, 56.6 mmol, 2.5 equivalents). The resulting mixture was stirred at 60 ° C for 12 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=2/1, v/v) to give tert-butyl 3-chloro-5-(hydroxymethyl)-1H-indole-1-carboxylate (4.3 g, 68%) as a white solid.

Step 4: Preparation of tert-butyl 3-chloro-5-(chloromethyl)-1H-indole-1-carboxylate

To a solution of tert-butyl 3-chloro-5-(hydroxymethyl)-1H-indole-1-carboxylate (1.5 g, 5.34 mmol, 1 equiv) in anhydrous DCM (30 mL) was added _Et3N (1.5 mL, 10.68 mmol, 2 equiv) and MsCl (0.62 mL, 8.01 mmol, 1.5 equiv). The resulting mixture was stirred at room temperature for 24 h and then quenched by adding water. The mixture was extracted with DCM. The combined extracts were dried and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=20/1, v/v) to give tert-butyl 3-chloro-5-(chloromethyl)-1H-indole-1-carboxylate (1.17 g, 73%) as a white solid.

Step 5: Preparation of tert-butyl 3-chloro-5-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-1H-indole-1-carboxylate

To a solution of tert-butyl 3-chloro-5-(chloromethyl)-1H-indole-1-carboxylate (1.1 g, 3.68 mmol, 1.0 equiv) in dioxane (20 mL) was added methyl 2-(trimethylstannyl)isonicotinate (1.22 g, 4.05 mmol, 1.1 equiv) and Pd( _PPh3 ) _{2Cl2 (} 260 mg, 0.37 mmol, 0.1 equiv). The mixture was stirred at 90°C for 3 h under a nitrogen atmosphere, concentrated, and purified by chromatography on a silica gel column (PE/EtOAc=20/1, v/v) to give tert-butyl 3-chloro-5-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-1H-indole-1-carboxylate (690 mg, 47%) as an off-white solid.

Step 6: Preparation of tert-butyl 5-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-3-methyl-1H-indole-1-carboxylate

To a solution of tert-butyl 3-chloro-5-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-1H-indole-1-carboxylate (400 mg, 1.0 mmol, 1 equivalent) in 1,4-dioxane (10 mL) was added potassium phosphate (424 mg, 2 mmol, 2 equivalents), trimethylboroxine (504 mg, 2 mmol, 2 equivalents, 50% purity in THF), tris(dibenzylideneacetone)dipalladium (46 mg, 0.05 mmol, 0.05 equivalents), and 2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl (X-Phos) (134 mg, 0.30 mmol, 0.30 equivalents) under argon, and the mixture was heated to 110° C. and stirred for 4 h. Ice-cold water was then added, and the mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried, and then concentrated under reduced pressure. The resulting residue was purified by chromatography on a silica gel column (EtOAc/PE=1/5, v/v) to give tert-butyl 5-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-3-methyl-1H-indole-1-carboxylate (280 mg, 74%) as a yellow solid.

Step 7: Preparation of 2-((3-methyl-1H-indol-5-yl)methyl)isonicotinic acid

To a solution of tert-butyl 5-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-3-methyl-1H-indole-1-carboxylate (280 mg, 0.74 mmol, 1 equiv) in DCM (3 mL) was added TFA (3 mL). The mixture was stirred at room temperature for 2 h and then concentrated. The residue was diluted with DCM and washed with saturated NaHCO _3. The organic layer was concentrated and the residue was dissolved in THF/H ₂ O (10 mL, 1: 1). LiOH.H ₂ O (47 mg, 1.11 mmol, 1.5 equiv) was added to the mixture. The mixture was stirred at room temperature for 1 h and the mixture was acidified to pH 3 with 1N HCl solution. The mixture was extracted with DCM, and the combined organic layers were dried and concentrated to give 2-((3-methyl-1H-indol-5-yl)methyl)isonicotinic acid (99 mg, 50%) as a yellow solid.

Intermediate 29: Preparation of 2-((2-(acetamidomethyl)quinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of 6-(methoxycarbonyl)quinoline 1-oxide

A mixture of quinoline-6-formyl ester (10 g, 53.5 mmol, 1 equivalent) and m-CPBA (18.4 g, 0.106 mol, 2 equivalents) in DCM (50 mL) was stirred at room temperature overnight. Saturated _NaHCO3 aqueous solution (40 mL) was added to the reaction mixture, and the resulting mixture was stirred for 30 min. The organic layer was separated, dried, filtered and concentrated. The resulting residue was recrystallized in EtOAc (5 mL) to obtain 6-(methoxycarbonyl)quinoline 1-oxide (8.0 g, 74%) as a light yellow solid.

Step 2: Preparation of methyl 2-chloroquinoline-6-carboxylate and methyl 4-chloroquinoline-6-carboxylate

Phosphorus oxychloride (20mL) is added to 6-(methoxycarbonyl)quinoline 1-oxide (4.0g, 19.7mmol, 1 equivalent).Then the resulting mixture is stirred at room temperature for 2h under _N .Then volatile matter is removed under vacuum, and the residue is dissolved in DCM.The mixture is washed with saturated NaHCO ₃ aqueous solution, dried and concentrated.The resulting residue is purified by chromatography on a silica gel column (PE/EtOAc=10/1, v/v) to obtain 2-chloroquinoline-6-methyl formate (1.2g, 28%) and 4-chloroquinoline-6-methyl formate (2.5g, 57%).

Step 3: Preparation of methyl 2-cyanoquinoline-6-carboxylate

To a suspension of methyl 2-chloroquinoline-6-carboxylate (1.2 g, 5.43 mmol, 1 equivalent) in DMF ( ₁₅ mL) was added Zn(CN) (1.11 g, 10.86 mmol, 2 equivalents) and Pd( _PPh ) ₍ 628 mg, 0.54 mmol, 0.1 equivalent). The reaction mixture was stirred at 100 ° C for 3 h under nitrogen. The cooled mixture was partitioned between EtOAc and water. The organic layer was separated and concentrated. The resulting residue was purified by chromatography on a silica gel column to give methyl 2-cyanoquinoline-6-carboxylate (980 mg, 85%) as a yellow solid.

Step 4: Preparation of 6-hydroxymethyl-quinoline-2-carbonitrile

To a suspension of methyl 2-cyanoquinoline-6-formate (980 mg, 4.62 mmol, 1 equivalent) in anhydrous THF (30 mL) was added LiAlH(t-BuO) ₃ (2.94 g, 11.56 mmol, 2.5 equivalents). The resulting mixture was stirred at 60 ° C for 12 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=1/2, v/v) to give 6-hydroxymethyl-quinoline-2-carbonitrile (722 mg, 83%) as a yellow solid.

Step 5: Preparation of 6-chloromethyl-quinoline-2-carbonitrile

To 6-hydroxymethyl-quinoline-2-carbonitrile (2.1 g, 11.41 mmol, 1 eq) was added _SOCl₂ (50 mL) and the mixture was stirred at room temperature for 3 h. The volatiles were then removed in vacuo at 40°C and the residue was dissolved in DCM. The mixture was washed with saturated _NaHCO₃ solution, dried and concentrated to give 6-chloromethyl-quinoline-2-carbonitrile (2.10 g, 91%) as a yellow solid. Step 6: Preparation of methyl 2-((2-cyanoquinolin-6-yl)methyl)isonicotinate

To a solution of 6-chloromethyl-quinoline-2-carbonitrile (2.10 g, 10.40 mmol, 1.0 equiv) in dioxane (50 mL) was added methyl 2-(trimethylstannyl)isonicotinate (3.44 g, 11.44 mmol, 1.1 equiv) and Pd(PPh ₃ ) ₂ Cl ₂ (730 mg, 1.04 mmol, 0.1 equiv). The mixture was stirred at 90° C. under nitrogen for 3 h, concentrated, and purified by chromatography on a silica gel column (DCM/MeOH=100/1, v/v) to give methyl 2-((2-cyanoquinolin-6-yl)methyl)isonicotinate (1.42 g, 45%) as a yellow solid.

Step 7: Preparation of methyl 2-((2-((tert-butoxycarbonylamino)methyl)quinolin-6-yl)methyl)isonicotinate

To a solution of methyl 2-((2-cyanoquinolin-6-yl)methyl)isonicotinate (1.0 g, 3.30 mmol, 1 eq) in MeOH (30 mL) was added Boc ₂ O (1.08 g, 5.00 mmol, 1.5 eq) and Raney nickel (200 mg). The mixture was stirred at room temperature for 5 h under hydrogen. The Raney nickel was removed by filtration, and the filtrate was concentrated. The resulting residue was purified by chromatography on a silica gel column (EtOAc/PE=1/2, v/v) to give methyl 2-((2-((tert-butoxycarbonylamino)methyl)quinolin-6-yl)methyl)isonicotinate (1.1 g, 82%) as a yellow solid.

Step 8: Preparation of methyl 2-((2-(acetamidomethyl)quinolin-6-yl)methyl)isonicotinate

To a solution of methyl 2-((2-((tert-butoxycarbonylamino)methyl)quinolin-6-yl)methyl)isonicotinate (1.1 g, 2.70 mmol, 1 equivalent) in EtOAc (5 mL) was added HCl/EtOAc solution (30 mL, 10 N). The mixture was stirred at room temperature for 2 h, and the precipitate was collected by filtration. To a suspension of the crude product obtained above in DCM (30 mL), TEA (818 mg, 8.10 mmol, 3 equivalents) and AcCl (316 mg, 4.05 mmol, 1.5 equivalents) were added. The mixture was stirred at room temperature for 1 h and the mixture was concentrated. The resulting residue was purified by chromatography on a silica gel column (EtOAc/PE=1/2, v/v) to give methyl 2-((2-(acetamidomethyl)quinolin-6-yl)methyl)isonicotinate (744 mg, 79% for 2 steps) as a yellow solid.

Step 9: Preparation of 2-[2-(acetylamino-methyl)-quinolin-6-ylmethyl]-isonicotinic acid

To a solution of methyl 2-((2-(acetamidomethyl)quinolin-6-yl)methyl)isonicotinate (744 mg, 2.13 mmol, 1 eq) in THF/H ₂ O (30 mL, 1:1) was added LiOH.H ₂ O (134 mg, 3.20 mmol, 1.5 eq). The mixture was stirred at room temperature for 1 h and acidified to pH 3 with 1N HCl solution. The mixture was extracted with DCM. The organic layer was washed with brine, then dried and concentrated to give 2-[2-(acetylamino-methyl)-quinolin-6-ylmethyl]-isonicotinic acid (649 mg, 91%) as a yellow solid, which was used without further purification.

Intermediate 30: Preparation of 2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinic acid

Step 1: Preparation of methyl 2-(methylsulfonyl)quinoline-6-carboxylate

A mixture of methyl 2-chloroquinoline-6-carboxylate (1.2 g, 5.43 mmol, 1 eq), sodium methanesulfinate (665 mg, 6.51 mmol, 1.2 eq), copper iodide (103 mg, 0.54 mol, 0.1 eq), and L-proline sodium salt (148 mg, 1.08 mol, 0.2 eq) in 110 mL of DMSO was heated to 110° C. under nitrogen and stirred for 15 h. The cooled mixture was partitioned between ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with brine, dried over MgSO ₄ , and concentrated in vacuo. The resulting residue was purified by chromatography on a silica gel column (EtOAc/PE=1/2, v/v) to give methyl 2-(methylsulfonyl)quinoline-6-carboxylate (830 mg, 58%) as a yellow solid.

Step 2: Preparation of (2-methanesulfonyl-quinolin-6-yl)-methanol

To a solution of methyl 2-(methylsulfonyl)quinoline-6-carboxylate (830 mg, 3.13 mmol, 1 equivalent) in THF (40 mL) was added LiAlH(t-BuO) ₃ (2.0 g, 7.83 mmol, 2.5 equivalents). The resulting mixture was stirred at 40 ° C for 12 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=1/1, v/v) to give (2-methylsulfonyl-quinolin-6-yl)-methanol (600 mg, 81%) as a yellow solid.

Step 3: Preparation of 6-chloromethyl-2-methanesulfonyl-quinoline

A mixture of (2-methanesulfonyl-quinolin-6-yl)-methanol (580 mg, 2.48 mmol, 1.0 equiv) in SOCl ₂ (10 mL) was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DCM and treated with saturated NaHCO ₃ solution. The organic layer was concentrated to give 6-chloromethyl-2-methanesulfonyl-quinoline (570 mg, 90%) as a yellow solid.

Step 4: Preparation of methyl 2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinate

To a solution of 6-chloromethyl-2-methanesulfonyl-quinoline (560 mg, 2.20 mmol, 1.0 equiv) in dioxane (15 mL) was added methyl 2-(trimethylstannyl)isonicotinate (727 mg, 2.42 mmol, 1.1 equiv) and _{Pd(PPh3)2Cl2 ( 154} mg, 0.22 mmol, 0.1 equiv). The mixture was stirred at 90°C under nitrogen for 3 h, concentrated, and purified by chromatography on a silica gel column (DCM/MeOH = 100/1, v/v) to give methyl 2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinate (140 mg, 18%) as a yellow solid.

Step 5: Preparation of 2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinic acid

To a solution of methyl 2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinate (140 mg, 0.39 mmol, 1 eq) in THF/H ₂ O (5 mL, 1:1) was added LiOH.H ₂ O (25 mg, 0.59 mmol, 1.5 eq). The mixture was stirred at room temperature for 1 h and acidified to pH 3 with 1 N HCl solution. The solvent was removed to give 2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinoic acid as a yellow solid, which was used in the next step without further purification.

Intermediate 31: Preparation of (3-chloro-4-fluoro-1H-indol-5-yl)methylamine hydrochloride

Step 1: Preparation of 4-fluoro-1-triisopropylsilyl-1H-indole

To a solution of 4-fluoro-1H-indole (5.0 g, 37.04 mmol, 1.0 equivalent) in anhydrous THF (200 mL) was added NaH solution (1.63 g, 40.74 mmol, 1.1 equivalents, 60% purity) at 0 ° C. After stirring for 0.5 h, TIPSCl (7.8 g, 40.74 mmol, 1.1 equivalents) was added. The mixture was then stirred at room temperature for 1 h and quenched by adding water. The mixture was extracted with EtOAc. The combined organic layers were dried and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=10/1, v/v) to obtain 4-fluoro-1-triisopropylsilyl-1H-indole (10.0 g, 84%) as a yellow oil.

Step 2: Preparation of 4-fluoro-1-(triisopropylsilyl)-1H-indole-5-carbaldehyde

To a solution of 4-fluoro-1-triisopropylsilyl-1H-indole (10 g, 34.36 mmol, 1.0 equiv) in THF (30 mL) was slowly added s-BuLi (27.5 mL, 41.24 mmol, 1.5 M, 1.2 equiv) at -78 ° C. The mixture was stirred for 1 h. DMF (7.52 g, 103 mmol, 3.0 equiv) was added dropwise. The mixture was stirred at -78 ° C for 1 h. The reaction was quenched with saturated aqueous NH ₄ Cl solution. The mixture was extracted with EtOAc (50 mL × 3). The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by chromatography on a silica gel column (PE / EtOAc = 100 / 1) to give 4-fluoro-1- (triisopropylsilyl) -1H-indole-5-carbaldehyde (6.6 g, 60%) as a yellow oil.

Step 3: Preparation of 4-fluoro-1H-indole-5-carbaldehyde oxime

A mixture of 4-fluoro-1-(triisopropylsilyl)-1H-indole-5-carbaldehyde (6.5 g, 20.4 mmol, 1.0 eq) and _NH OH.HCl (2.82 g, 40.8 mmol, 2.0 eq) in NH ₃ /MeOH (15% w/w, 200 mL) was stirred at room temperature overnight. The mixture was concentrated. The residue was purified by chromatography on a silica gel column (PE/EtOAc=50/1) to give 4-fluoro-1H-indole-5-carbaldehyde oxime (3.6 g, crude) as a yellow solid.

Step 4: Preparation of (4-fluoro-1H-indol-5-yl)methanamine

A mixture of 4-fluoro-1H-indole-5-carbaldehyde oxime (3.6 g, 20.2 mmol, 1.0 equiv) and Raney nickel (600 mg) in NH ₃ /MeOH (15% w/w, 200 mL) was stirred under H ₂ atmosphere (1 atm) at room temperature overnight. The mixture was filtered and concentrated to give (4-fluoro-1H-indol-5-yl)methanamine (3.0 g, 92%) as a gray solid, which was used in the next step without further purification.

Step 5: Preparation of tert-butyl (4-fluoro-1H-indol-5-yl)methylcarbamate

To a solution of (4-fluoro-1H-indol-5-yl)methanamine (3.0 g, 18.3 mmol, 1.0 equiv) in DCM (150 mL) was added TEA (2.22 g, 22.0 mmol, 1.2 equiv) and Boc ₂ O (4.80 g, 22.0 mmol, 1.2 equiv). The mixture was stirred at room temperature for 2 h and concentrated. The residue was purified by chromatography on a silica gel column (PE/EtOAc=10/1, v/v) to give tert-butyl (4-fluoro-1H-indol-5-yl)methylcarbamate (4.2 g, 87%) as a yellow solid.

Step 6: Preparation of tert-butyl (3-chloro-4-fluoro-1H-indol-5-yl)methylcarbamate

To a solution of tert-butyl (4-fluoro-1H-indol-5-yl)methylcarbamate (4.2 g, 15.9 mmol, 1.0 equiv) in DCM (150 mL) was added NCS (2.22 g, 16.7 mmol, 1.05 equiv). The mixture was stirred at room temperature for 1 h and concentrated. The residue was purified by chromatography on a silica gel column (PE/EtOAc=10/1, v/v) to give tert-butyl (3-chloro-4-fluoro-1H-indol-5-yl)methylcarbamate (4.0 g, 85%) as a yellow solid.

Step 7: Preparation of (3-chloro-4-fluoro-1H-indol-5-yl)methylamine hydrochloride

To a solution of tert-butyl (3-chloro-4-fluoro-1H-indol-5-yl)methylcarbamate (4.0 g, 13.4 mmol, 1.0 equiv) in EtOAc (20 mL) was added a solution of HCl in EtOAc (10 N, 80 mL). The mixture was stirred at room temperature for 1 h, and the precipitate was collected by filtration to give (3-chloro-4-fluoro-1H-indol-5-yl)methanamine hydrochloride (2.57 g, 82%) as a yellow solid.

Intermediate 32: Preparation of (3-chloro-6-fluoro-1H-indol-5-yl)methylamine hydrochloride

Step 1: Preparation of 6-fluoro-1-triisopropylsilyl-1H-indole

To a solution of 6-fluoro-1H-indole (5.0 g, 37.04 mmol, 1.0 equivalent) in anhydrous THF (200 mL) was added a solution of NaH (1.63 g, 40.74 mmol, 1.1 equivalents, 60% purity) in THF (50 mL) at 0 ° C. After stirring for 0.5 h, TIPSCl (7.8 g, 40.74 mmol, 1.1 equivalents) was added. The mixture was stirred at room temperature for 1 h and quenched by adding water. The mixture was extracted with EtOAc. The combined organic layers were dried and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=10/1, v/v) to obtain 6-fluoro-1-triisopropylsilyl-1H-indole (10.3 g, 87%) as a yellow oil.

Step 2: Preparation of 6-fluoro-1-(triisopropylsilyl)-1H-indole-5-carbaldehyde

To a solution of 6-fluoro-1-(triisopropylsilyl)-1H-indole (2 g, 6.9 mmol, 1.0 equiv) in THF (30 mL) was slowly added s-BuLi (6.3 mL, 1.3 M, 1.2 equiv) at -78 ° C. The mixture was stirred at this temperature for 1 h. DMF (1.5 g, 20.7 mmol, 3.0 equiv) was added dropwise. The mixture was stirred at -78 ° C for 1 h. The reaction was quenched with saturated aqueous NH ₄ Cl solution. The obtained mixture was extracted with EtOAc (50 mL×3). The organic layers were combined and washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=100/1) to give 6-fluoro-1-(triisopropylsilyl)-1H-indole-5-carbaldehyde (950 mg, 57%) as a yellow oil.

Step 3: Preparation of 6-fluoro-1H-indole-5-carbaldehyde oxime

A mixture of 6-fluoro-1-(triisopropylsilyl)-1H-indole-5-carbaldehyde (780 mg, 2.45 mmol, 1.0 eq) and _NH OH.HCl (340 mg, 4.89 mmol, 2.0 eq) in NH ₃ /MeOH (15% w/w, 10 mL) was stirred at room temperature overnight. The mixture was concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=50/1) to give 6-fluoro-1H-indole-5-carbaldehyde oxime (460 mg, crude) as a yellow solid.

Step 4: Preparation of (6-fluoro-1H-indol-5-yl)methanamine

A mixture of 6-fluoro-1H-indole-5-carbaldehyde oxime (460 mg, 1.38 mmol, 1.0 equiv) and Raney nickel (100 mg) in NH ₃ /MeOH (15% w/w, 10 mL) was stirred at room temperature under an atmosphere of H ₂ (1 atm) overnight. The mixture was filtered and concentrated to give (6-fluoro-1H-indol-5-yl)methanamine (420 mg, 95%) as a gray solid, which was used in the next step without further purification.

Step 5: Preparation of tert-butyl (6-fluoro-1H-indol-5-yl)methylcarbamate

To a solution of (6-fluoro-1H-indol-5-yl)methanamine (420 mg, 2.56 mmol, 1.0 equiv) in DCM (25 mL) was added TEA (0.43 mL, 3.07 mmol, 1.2 equiv) and Boc ₂ O (670 mg, 3.07 mmol, 1.2 equiv). The mixture was stirred at room temperature for 2 h and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=10/1, v/v) to give tert-butyl (6-fluoro-1H-indol-5-yl)methylcarbamate (608 mg, 90%) as a yellow solid.

Step 6: Preparation of tert-butyl (3-chloro-6-fluoro-1H-indol-5-yl)methylcarbamate

To a solution of tert-butyl (6-fluoro-1H-indol-5-yl)methylcarbamate (600 mg, 2.27 mmol, 1.0 equiv) in DCM (20 mL) was added NCS (317 mg, 2.39 mmol, 1.05 equiv). The mixture was stirred at room temperature for 1 h and concentrated. The resulting residue was purified by chromatography on a silica gel column (PE/EtOAc=10/1, v/v) to give tert-butyl (3-chloro-6-fluoro-1H-indol-5-yl)methylcarbamate (547 mg, 81%) as a yellow solid.

Step 7: Preparation of (3-chloro-6-fluoro-1H-indol-5-yl)methanamine hydrochloride

To a solution of tert-butyl (3-chloro-6-fluoro-1H-indol-5-yl)methylcarbamate (547 mg, 1.84 mmol, 1.0 eq) in EtOAc (5 mL) was added a solution of HCl in EtOAc (10 N, 10 mL). The mixture was stirred at room temperature for 1 h, and the precipitate was collected by filtration to give (3-chloro-6-fluoro-1H-indol-5-yl)methylamine hydrochloride as a yellow solid.

Example 1: Preparation of 6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide

6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide

[0266] To a solution of N-(3-chloro-6-fluoro-1H-indol-5-ylmethyl)-2-(2-cyano-quinolin-6-ylmethyl)-isonicotinamide (synthesized as described in Example 3, 70 mg, 0.15 mmol, 1.0 equiv) in MeOH (1.5 mL)/ _H2O (0.6 mL) was added ammonium hydroxide (2.1 mL) and hydrogen peroxide (0.1 mL). The mixture was stirred at 30°C for 3 h. The mixture was concentrated in vacuo, and the residue was purified by preparative HPLC to afford 6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide (18 mg, 25%) as a white solid. LRMS (M+H ⁺ ) m/z calcd 488.1, found 487.8.

¹ H NMR (DMSO-d ₆ , 400MHz) δ11.41 (s, 1H), 9.27 (s, 1H), 8.67 (d, 1H), 8.49 (d, 1H), 8.26 (s, 1H), 8.13 (d, 1H), 8.01 (d, 1H), 7.95 (s , 1H), 7.82(s, 1H), 7.78(d, 2H), 7.67(d, 1H), 7.51(s, 1H), 7.46(d, 1H), 7.24(d, 1H), 4.59(d, 2H), 4.40(s, 2H).

Example 2: Preparation of 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide

6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide

6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide (18 mg, 19%) was prepared as a white solid as described for 6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide (Example 1). LRMS (M+H ⁺ ) m/z calcd 441.2, found 440.9.

¹ H NMR (DMSO-d ₆ ,400MHz)δ8.66-8.61(m,2H),8.49(d,1H),8.27(s,1H),8.13(d,1H),8.06(d,1H),7.94(s,1H),7.80(s,1H ),7.78(s,1H),7.62(d,1H),6.13(s,1H),5.71(s,2H),4.38(s,1H),4.35(d,2H),2.30(s,3H),2.17(s,3H).

Example 3: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-cyanoquinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(2-cyano-quinolin-6-ylmethyl)-isonicotinic acid (250 mg, 0.86 mmol, 1.0 equiv) in DMF (10 mL) was added 6-aminomethyl-isoquinolin-1-ylamine (148.7 mg, 0.86 mmol, 1.0 equiv), followed by EDCI (280.7 mg, 1.46 mmol, 1.7 equiv), HOBT (174.2 mg, 1.29 mmol, 1.5 equiv), and TEA (0.47 mL, 3.4 mmol, 4.0 equiv). The reaction mixture was heated to 45° C. and stirred overnight. Water was added, and the mixture was extracted with DCM. The organic layer was washed with water, dried over Na ₂ SO ₄ , filtered, and concentrated. The residue was purified by preparative HPLC to give N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-cyanoquinolin-6-yl)methyl)isonicotinamide (95 mg, 25%) as a yellow solid. LRMS (M+H ⁺ ) m/z calcd 445.2, found 445.2.

¹ H NMR (DMSO-d ₆ ,400MHz): δ9.41(t,1H),8.58(d,1H),8.62(d,1H),8.15(d,1H),8.09(d,1H),8.01(t,2H),7.91(d,1H),7.89(d,1H),7 .84(s,1H),7.77(d,1H),7.70(dd,1H),7.56(s,1H),7.41(dd,1H),6.85(d,1H),6.76(s,2H),4.62(d,2H),4.44(s,2H).

Example 4: Preparation of 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide

6-((4-((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide

6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide (17 mg, 23%) was prepared as a white solid as described for 6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-2-carboxamide (Example 1). LRMS (M+H ⁺ ) m/z calcd 463.2, found 462.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.43(t,1H),8.69(d,1H),8.51(d,1H),8.23(s,1H),8.16(t,2H),8.07(d,1H),7.97(s,1H),7.8 4-7.76(m,4H),7.71(d,1H),7.57(s,1H),7.43(d,1H),6.86(d,1H),6.80(s,2H),4.63(d,2H),4.42(s,2H).

Example 5: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(3-isocyano-quinolin-6-ylmethyl)-isonicotinic acid (100 mg, 0.346 mmol, 1.0 eq) and (3-chloro-6-fluoro-1H-indol-5-yl)-methylamine (81 mg, 0.346 mmol, 1.0 eq) in DMF (8 mL) was added HOBT (70 mg, 0.519 mmol, 1.5 eq), EDCI (99.5 mg, 0.519 mmol, 1.5 eq) and _Et3N (140 mg, 1.384 mmol, 4 eq). The mixture was stirred at room temperature for 15 h and diluted with water. The organic layer was separated and the aqueous layer was extracted with DCM. The combined extracts were dried and concentrated. The residue was purified by preparative HPLC to give N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide (15 mg, 9%) as an off-white solid. LRMS (M+H ⁺ ) m/z calcd 470.1, found 470.1.

¹ H NMR (DMSO-d ₆ , 400MHz) δ11.37 (s, 1H), 9.23 (t, 1H), 9.07 (d, 1H), 8.99 (s, 1H), 8.61 (d, 1H), 8.02 (d, 1H), 7.93 (s, 1H), 7.88 (d, 1H), 7.77 (s, 1H), 7.62 (d, 1H), 7.47 (d, 1H), 7.40 (d, 1H), 7.18 (d, 1H), 4.54 (d, 2H), 4.37 (s, 2H).

Example 6: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide (58 mg, 23.7%) was prepared as an off-white solid as described for N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide (Example 5). LRMS (M+H ⁺ ) m/z calcd 445.2, found 444.9.

¹ H NMR (DMSO-d ₆ ,400MHz)δ13.20-13.12(m,1H),9.51(t,1H),9.12(d,1H),9.06-8.99(m,2H),8.69(d,1H),8.50(d,1H),8.07(d,1H ),7.99(s,1H),7.93(dd,1H),7.83(s,2H),7.73-7.71(m,2H),7.66(d,1H),7.22(d,1H),4.68(d,2H),4.44(s,2H).

Example 7: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide (45 mg, 32.3%) was prepared as an off-white solid as described for N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-cyanoquinolin-6-yl)methyl)isonicotinamide (Example 5). LRMS (M+H ⁺ ) m/z calcd 423.2, found 422.9. ¹ H NMR (DMSO-d ₆ ,300MHz)δ9.12(s,1H),9.03(s,1H),8.69-8.65(m,1H),8.61-8.59(m,1H),8.05(dd,1H),7.96-7.89(m,2H), 7.77(s,1H),7.61-7.60(m,1H),6.16(d,1H),5.91-5.87(m,2H),4.38-4.33(m,4H),2.31(s,3H),2.18(s,3H).

Example 8: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((7-fluoroquinoxalin-2-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((7-fluoroquinoxalin-2-yl)methyl)isonicotinamide

Step 1: Preparation of 7-fluoro-3,4-dihydro-1H-quinoxalin-2-one

To a solution of 4-fluoro-benzene-1,2-diamine (20 g, 0.159 mol, 1 eq) in DMF (150 mL) was added _Et₃N (44 mL, 0.318 mol, 2 eq) followed by ethyl 2-bromoacetate (29 g, 0.175 mol, 1.1 eq). The reaction mixture was stirred at room temperature for 16 h and then at 80°C for 3 h. The DMF was evaporated by distillation. The reaction mixture was partitioned between _H₂O and EtOAc. The organic layer was washed with saturated _NaHCO₃ , brine, and dried _{over Na₂SO₄} . The solvent was evaporated under reduced pressure. The desired product was precipitated in a mixture _{of CH₂Cl₂} and hexane (1:1 ratio). The mixture was filtered, and the filtrate was concentrated to dryness to give 7-fluoro-3,4-dihydro-1H-quinoxalin-2-one (22 g, 83%).

Step 2: Preparation of 7-fluoro-quinoxalin-2-ol

A mixture of 7-fluoro-3,4-dihydro-1H-quinoxaline-2-one (4.0 g, 24 mmol, 1.0 equivalent), sodium hydroxide (1.93 g, 48 mmol, 2.0 equivalent) and 3% hydrogen peroxide solution (50 mL) was refluxed for 2 h and then acidified by slowly adding acetic acid. The resulting mixture was cooled to room temperature. The precipitated solid was collected by filtration, washed with ice water, and dried in vacuo. The resulting residue was purified by column chromatography (DCM/MeOH=50:1, v/v) to obtain 7-fluoro-quinoxaline-2-ol (2.60 g, 69%).

Step 3: Preparation of 2-chloro-7-fluoro-quinoxaline

To a suspension of 7-fluoro-quinoxaline-2-ol (2 g, 12 mmol, 1 equiv) in pure phosphorus oxychloride (10 mL) was added DMF (2 drops). The mixture was heated to 100° C. for 3 h. It was then allowed to cool to room temperature. The phosphorus oxychloride was removed in vacuo and the residue was dissolved in EtOAc and dropped into ice water with stirring. The mixture was extracted three times with EtOAc and the combined organic layers were washed with saturated NaHCO ₃ solution. The organic layer was concentrated to give 2-chloro-7-fluoro-quinoxaline (1.7 g, 77%). Step 4: Preparation of 7-fluoro-2-(trimethylstannyl)quinoxaline

To a solution of 2-chloro-7-fluoro-quinoxaline (2.0 g, 11 mmol, 1 eq) in toluene (50 mL) was added hexamethylditin (7.2 g, 22 mmol, 2 eq) and Pd(PPh ₃ ) ₄ (635 mg, 0.55 mmol, 0.05 eq). The mixture was stirred at 90° C. under nitrogen for 12 h. The reaction mixture was concentrated and the resulting residue was used in the next step without further purification.

Step 5: Preparation of methyl 2-((7-fluoroquinoxalin-2-yl)methyl)isonicotinate

To a solution of 7-fluoro-2-trimethylstannyl-quinoxaline (3.43 g, 11 mmol, 1.0 equiv) in dioxane (60 mL) was added methyl 2-(bromomethyl)isonicotinate (2.5 g, 11 mmol, 1.0 equiv) and Pd(PPh ₃ ) ₂ Cl ₂ (386 mg, 0.55 mmol, 0.05 equiv). The mixture was stirred at 90° C. under a nitrogen atmosphere for 3 h, concentrated, and purified by silica gel chromatography (PE/EtOAc=3/1, v/v) to give methyl 2-((7-fluoroquinoxalin-2-yl)methyl)isonicotinate (300 mg, 9% over 2 steps) as an off-white solid.

Step 6: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((7-fluoroquinoxalin-2-yl)methyl)isonicotinamide

To a solution of methyl 2-((7-fluoroquinoxalin-2-yl)methyl)isonicotinate (70 mg, 0.235 mmol, 1.0 equiv) in THF (10 mL)/ _H₂O (2 mL) was added NaOH (11.3 mg, 0.282 mmol, 1.2 equiv). The mixture was stirred at 45° C. for 2 h and acidified to pH 5-6 with 1N HCl solution. The mixture was concentrated in vacuo, and the resulting residue was used in the next step without further purification. To a solution of this crude product and 6-aminomethyl-isoquinolin-1-ylamine (55.2 mg, 0.235 mmol, 1.0 equiv) in DMF (8 mL) were added HOBT (47.6 mg, 0.352 mmol, 1.5 equiv), EDCI (67.5 mg, 0.352 mmol, 1.5 equiv), and _Et₃N (95.1 mg, 0.940 mmol, 4 equiv). The mixture was stirred at 45° C. for 15 h and then concentrated. The resulting residue was purified by preparative HPLC to give N-((1-aminoisoquinolin-6-yl)methyl)-2-((7-fluoroquinoxalin-2-yl)methyl)isonicotinamide (15 mg, 14.6%) as a yellow solid. LRMS (M+H ⁺ ) m/z calcd 439.2, found 438.8. ¹ H NMR (CD ₃ OD, 400 MHz) δ 8.79 (s, 1H), 8.50 (d, 1H), 7.95-7.89 (m, 2H), 7.77 (s, 1H), 7.58-7.47 (m, 5H), 7.36 (d, 1H), 6.79 (d, 1H), 4.60 (s, 2H), 4.51 (s, 2H)

Example 9: Preparation of 2-[2-(acetylamino-methyl)-quinolin-6-ylmethyl]-N-(3-chloro-6-fluoro-1H-indol-5-ylmethyl)-isonicotinamide

2-[2-(Acetylamino-methyl)-quinolin-6-ylmethyl]-N-(3-chloro-6-fluoro-1H-indol-5-ylmethyl)-isonicotinamide

To a solution of methyl 2-((2-(acetamidomethyl)quinolin-6-yl)methyl)isonicotinate (100 mg, 0.287 mmol, 1.0 equiv) in THF (10 mL)/H ₂ O (2 mL) was added LiOH·H ₂ O (14.4 mg, 0.344 mmol, 1.2 equiv). The mixture was stirred at 45° C. for 2 h and acidified to pH 5-6 with 1N HCl solution. The mixture was concentrated in vacuo and the residue was used directly without further purification. To this crude product and (3-chloro-6-fluoro-1H-indol-5-yl)-methylamine (67.3 mg, 0.287 mmol, 1.0 eq) in DMF (8 mL) was added HOBT (58 mg, 0.430 mmol, 1.5 eq), EDCI (82.4 mg, 0.430 mmol, 1.5 eq) and _Et3N (116 mg, 1.146 mmol, 4 eq). The mixture was stirred at 45°C for 15 h and then concentrated. The resulting residue was purified by preparative HPLC to give 2-[2-(acetylamino-methyl)-quinolin-6-ylmethyl]-N-(3-chloro-6-fluoro-1H-indol-5-ylmethyl)-isonicotinamide (15 mg, 10.2%) as a yellow solid. LRMS (M+H ⁺ ) m/z calculated value 516.2, found value 515.8. ¹ H NMR (DMSO-d ₆ ,400MHz)δ11.43(s,1H),9.30(t,1H),9.01(s,1H),8.69-8.64(m,2H),8.48(d,1H),7.98(d,1H),7.93(s,1H),7.84- 7.78(m,2H),7.71(d,1H),7.56-7.51(m,3H),7.45(d,1H),7.23(d,1H),4.60-4.56(m,4H),4.41(s,2H),1.94(s,3H).

Example 10: Preparation of 2-[2-(acetylamino-methyl)-quinolin-6-ylmethyl]-N-(1-amino-isoquinolin-6-ylmethyl)-isonicotinamide

2-[2-(Acetylamino-methyl)-quinolin-6-ylmethyl]-N-(1-amino-isoquinolin-6-ylmethyl)-isonicotinamide

2-[2-(Acetylamino-methyl)-quinolin-6-ylmethyl]-N-(1-amino-isoquinolin-6-ylmethyl)-isonicotinamide (15 mg, 10.7%) was prepared as an off-white solid as described for 2-[2-(acetylamino-methyl)-quinolin-6-ylmethyl]-N-(3-chloro-6-fluoro-1H-indol-5-ylmethyl)-isonicotinamide (Example 9). LRMS (M+H ⁺ ) m/z calcd 491.2, found 490.9. ¹ H NMR (DMSO-d ₆ ,400MHz)δ8.97(d,1H),8.76(d,1H),8.41(d,1H),8.24-8.20(m,2H),8.08(dd,1H),7.99(s,1H),7.94-7 .87(m,3H),7.78(dd,1H),7.57(d,1H),7.21(d,1H),4.88(s,2H),4.82(s,2H),4.63(s,2H),2.13(s,3H).

Example 11: Preparation of 2-[2-(acetylamino-methyl)-quinolin-6-ylmethyl]-N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-isonicotinamide

2-[2-(Acetylamino-methyl)-quinolin-6-ylmethyl]-N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-isonicotinamide

2-[2-(Acetylamino-methyl)-quinolin-6-ylmethyl]-N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-isonicotinamide (10 mg, 7.46%) was prepared as a yellow solid as described for 2-[2-(acetylamino-methyl)-quinolin-6-ylmethyl]-N-(3-chloro-6-fluoro-1H-indol-5-ylmethyl)-isonicotinamide (Example 9). LRMS (M+H ⁺ ) m/z calcd 469.2, found 469.2. ¹ H NMR (DMSO-d ₆ , 400MHz) δ8.48 (m, 1H), 8.13 (d, 1H), 7.82 (d, 1H), 7.66 (s, 1H), 7.59-7.54 (m, 2H), 7.48 (dd, 1H), 7.3 5(d, 1H), 6.18(s, 1H), 4.53(s, 2H), 4.37(s, 2H), 4.26(s, 2H), 2.27(s, 3H), 2.13(s, 3H), 1.96(s, 3H).

Example 12: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide

Step 1: Preparation of 6-methylsulfonyl-quinoline

A mixture of 6-bromo-quinoline (20.7 g, 0.1 mol, 1 equiv), sodium methanesulfinate (12.2 g, 0.12 mol, 1.2 equiv), copper iodide (1.9 g, 0.01 mol, 0.1 equiv), and L-proline sodium salt (2.74 g, 0.02 mol, 0.2 equiv) in 200 mL of DMSO was heated to 110° C. under nitrogen for 15 h. The cooled mixture was partitioned between ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with brine, dried over MgSO ₄ , and concentrated in vacuo. The resulting residue was purified by silica gel column (EtOAc/PE=1/2, v/v) to give 6-methanesulfonyl-quinoline (13.5 g, 65%) as a yellow solid.

Step 2: Preparation of 3-bromo-6-methanesulfonyl-quinoline

To a mixture of 6-methylsulfonyl-quinoline (6.0 g, 29.0 mmol, 1 equivalent) and pyridine (4.7 mL, 58.0 mmol, 2 equivalents) in _CCl (250 mL) was added _Br (0.9 mL, 34.8 mmol, 1.2 equivalents) dropwise. The mixture was heated to reflux for 2 h and then cooled to room temperature. The liquid in the flask was poured out and washed with saturated aqueous _NaHCO and water. The dark solid at the bottom of the flask was distributed between aqueous _NaHCO and dichloromethane. The combined organic layers were washed again with water, dried, and then evaporated to dryness in vacuo. The crude product was purified by silica gel column (EtOAc/PE=1/10, v/v) to give 3-bromo-6-methylsulfonyl-quinoline (6.2 g, 75%) as a yellow solid.

Step 3: Preparation of 6-methylsulfonyl-3-vinyl-quinoline

To a solution of 3-bromo-6-methanesulfonyl-quinoline (2.9 g, 10.2 mmol, 1 eq) and vinylboronic acid pinacol cyclic ester (2.1 g, 12.2 mmol, 1.2 eq) in dioxane (50 mL) and water (10 mL) was added Na ₂ CO ₃ (3.24 g, 30.6 mmol, 3 eq) and Pd (dppf) Cl ₂ .CH ₂ Cl ₂ (833 mg, 1.02 mmol, 0.1 eq). The mixture was stirred at 95 ° C for 3 h. After cooling to room temperature, the solvent was removed in vacuo. The residue was purified by flash chromatography on a silica gel column (EtOAc / PE = 1 / 10, v / v) to give 6-methanesulfonyl-3-vinyl-quinoline (2.1 g, 88%) as a yellow solid.

Step 4: Preparation of 6-methanesulfonyl-quinoline-3-carbaldehyde

6- methanesulfonyl -3- vinyl - quinoline (2.1g, 9.0mmol, 1 equivalent) and dichloromethane (40mL) are added into 3 neck round-bottom flasks and are cooled to -78 DEG C. Ozone is bubbled into the reaction mixture until it is continuously blue (30min). The reaction mixture is sparged with oxygen until the blue fades, and quenched with dimethyl sulfide (6mL). The mixture is stirred at room temperature for 1h, is then concentrated, and is purified by flash column chromatography (EtOAc/PE=1/8, v/v) to obtain 6- methanesulfonyl -3- vinyl - quinoline -3- carbaldehyde (1.0g, 47%) as a white solid.

Step 5: Preparation of (6-methanesulfonyl-quinolin-3-yl)-methanol

To a solution of 6- methanesulfonyl-quinoline-3-carboxaldehyde (1.0 g, 4.25 mmol, 1 equivalent) in anhydrous MeOH (20 mL) was added NaBH ₄ (162 mg, 4.25 mmol, 1 equivalent) at 0° C. The mixture was stirred for 10 min at the same temperature. The reaction was quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The residue was purified by chromatography on a silica gel column (EtOAc/PE=1/2, v/v) to give (6- methanesulfonyl-quinoline-3-yl)-methanol (290 mg, 29%) as a yellow solid.

Step 6: Preparation of 3-chloromethyl-6-methylsulfonyl-quinoline

To (6-methylsulfonyl-quinolin-3-yl)-methanol (290 mg, 1.22 mmol, 1.0 equiv) was added _SOCl₂ (5 mL), and the mixture was stirred at room temperature for 2 h. The volatiles were then removed in vacuo at 40°C, and the residue was dissolved in DCM. The mixture was washed with saturated aqueous _NaHCO₃ , dried, and concentrated to give 3-chloromethyl-6-methylsulfonyl-quinoline (310 mg, 99%) as a yellow solid.

Step 7: Preparation of methyl 2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinate

To a solution of 3-chloromethyl-6-methanesulfonyl-quinoline (310 mg, 2.61 mmol, 1.0 equiv) in dioxane (20 mL) was added methyl 2-(trimethylstannyl)isonicotinate (864 mg, 2.87 mmol, 1.1 equiv) and Pd( _PPh3 ) _2Cl2 (183 mg, 0.26 mmol, 0.1 equiv). The mixture was stirred at 90°C for 3 h under a nitrogen atmosphere, concentrated, and purified by silica _gel chromatography (DCM/MeOH=100/1, v/v) to give methyl 2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinate (290 mg, 67%) as a yellow solid.

Step 8: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide

To a solution of methyl 2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinate (85 mg, 0.24 mmol, 1.0 equiv) in THF (3 mL)/H ₂ O (2 mL) was added LiOH.H ₂ O (15 mg, 0.36 mmol, 1.5 equiv). The mixture was stirred at 40° C. for 1 h and acidified to pH 5 with 1N HCl solution. The mixture was concentrated in vacuo, and the residue was used without further purification. To a solution of the above crude product and (3-chloro-6-fluoro-1H-indol-5-yl)-methylamine hydrochloride (68 mg, 0.29 mmol, 1.2 equiv) in DMF (5 mL) was added HATU (137 mg, 0.36 mmol, 1.5 equiv) and Et ₃ N (97 mg, 0.96 mmol, 4 equiv). The mixture was stirred at room temperature for 1 h and concentrated. The residue was purified by preparative HPLC to give N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinoamide (53 mg, 43% over 2 steps) as an off-white solid. LRMS (M+H ⁺ ) m/z calcd 523.1, found 522.8. ^1H NMR(DMSO-d6,400MHz)δ11.90(s,1H),9.29(s,1H),9.10(s,1H),8.66(d,1H),8.63(s,1H),8.50(s,1H),8.22(d ,1H),8.14(d,1H),7.85(s,1H),7.68(d,1H),7.52(s,1H),7.47(d,1H),4.61(d,2H),4.46(s,2H),3.37(s,3H).

Example 13: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide (56 mg, 46% over 2 steps) was prepared as an off-white solid as described for N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide (Example 12). LRMS (M+H ⁺ ) m/z calcd 498.2, found 497.9.

¹ H NMR (CD ₃ OD,400MHz)δ8.85(s,1H),8.51(d,1H),8.37(s,1H),8.21(s,1H),7.99(s,1H),7.89(d,1H),7.71(s,1H), 7.55(s,1H),7.53(d,1H),7.45(s,1H),7.32(d,1H),6.74(d,1H),4.56(s,2H),4.32(s,2H),3.07(s,3H).

Example 14: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((6-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide (Example 12) was prepared as an off-white solid (35 mg, 32% over 2 steps). LRMS (M+H ⁺ ) m/z calcd 476.2, found 476.0.

¹ H NMR (CD ₃ OD,400MHz)δ8.91(d,1H),8.51(d,1H),8.47(s,1H),8.31(s,1H),8.09(d,1H),8.08(d,1H),7.67( s,1H),7.51(d,1H),6.20(s,1H),4.40(s,2H),4.37(s,2H),3.11(s,3H),2.29(s,3H),2.16(s,3H).

Example 15: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of methyl 4-amino-3-iodobenzoate

To a solution of 4-aminobenzoic acid methyl ester (20g, 0.132mol, 1 equivalent) in AcOH (500mL) was added a solution of ICl (23.6g, 0.146mol, 1.1 equivalents) in AcOH (500mL) at 0°C. The mixture was stirred at room temperature for 2h. AcOH was evaporated under reduced pressure. The residue was diluted with DCM and washed with saturated NaHCO _3. The aqueous layer was extracted with DCM, and the combined extracts were dried and concentrated. The residue was purified by chromatography on a silica gel column (EtOAc/PE=1/15, v/v) to obtain 4-amino-3-iodobenzoic acid methyl ester (27.4g, 75%) as an off-white solid.

Step 2: Preparation of methyl 8-iodo-3-methylquinoline-6-carboxylate

A mixture of methyl 4-amino-3-iodobenzoate (26 g, 93.5 mmol), 2-methyl-acrolein (24.5 g, 0.28 mol, 3 equivalents) and 6N HCl (95 mL) was heated to reflux for 24 h. The mixture was then cooled and adjusted to pH 5-6 with NaHCO ₃ (aqueous solution). The mixture was extracted with DCM. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered, then concentrated, and purified by column chromatography (EtOAc/PE=1/20, v/v) to give methyl 8-iodo-3-methylquinoline-6-carboxylate (10.2 g, 33%) as a yellow solid.

Step 3: Preparation of (8-iodo-3-methyl-quinolin-6-yl)-methanol

To a solution of 8-iodo-3-methylquinoline-6-carboxylic acid methyl ester (7.5 g, 22.9 mmol, 1 equivalent) was added LiAlH(t-BuO) ₃ (14.6 g, 57.3 mmol, 2.5 equivalents). The resulting mixture was stirred at 40 ° C for 12 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=2/1, v/v) to give (8-iodo-3-methyl-quinolin-6-yl)-methanol (6.5 g, 95%) as a yellow solid.

Step 4: Preparation of (8-methanesulfonyl-3-methyl-quinolin-6-yl)-methanol

A mixture of (8-iodo-3-methyl-quinolin-6-yl)-methanol (6.5 g, 21.7 mmol, 1 eq), sodium methanesulfinate (2.66 g, 26.1 mmol, 1.2 eq), copper iodide (412 mg, 2.17 mol, 0.1 eq), and L-proline sodium salt (594 mg, 4.34 mol, 0.2 eq) in 100 mL of DMSO was heated to 110° C. under nitrogen for 15 h. The cooled mixture was partitioned between ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with brine, dried over _MgSO , and concentrated in vacuo. The residue was purified by silica gel column (EtOAc/PE=1/2, v/v) to give (8-methanesulfonyl-3-methyl-quinolin-6-yl)-methanol (3.3 g, 60%) as a yellow solid.

Step 5: Preparation of 6-chloromethyl-8-methylsulfonyl-3-methyl-quinoline

To (8-methylsulfonyl-3-methyl-quinolin-6-yl)-methanol (3.3 g, 13.1 mmol, 1.0 equiv) was added _SOCl₂ (50 mL) and the mixture was stirred at room temperature for 1 h. The volatiles were removed under vacuum and the residue was dissolved in DCM. The mixture was washed with saturated aqueous _NaHCO₃ , dried and concentrated to give 6-chloromethyl-8-methylsulfonyl-3-methyl-quinoline (3.4 g, 96%) as a yellow solid.

Step 6: Preparation of methyl 2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinate

To a solution of 6-chloromethyl-8-methanesulfonyl-3-methyl-quinoline (3.0 g, 11.1 mmol, 1.0 equiv) in dioxane (60 mL) was added methyl 2-(trimethylstannyl)isonicotinate (3.70 g, 12.3 mmol, 1.1 equiv) and Pd(PPh ₃ ) ₂ Cl ₂ (779 mg, 1.11 mmol, 0.1 equiv). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, concentrated, and purified by silica gel chromatography (DCM/MeOH=100/1, v/v) to give methyl 2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinate (2.26 g, 55%) as a yellow solid.

Step 7: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

To a solution of methyl 2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinate (120 mg, 0.32 mmol, 1.0 equiv) in THF (3 mL)/H ₂ O (2 mL) was added LiOH.H ₂ O (26.88 mg, 0.64 mmol, 2.0 equiv). The mixture was stirred at 40° C. for 1 h and acidified to pH 5 with 1 N HCl solution. The mixture was concentrated in vacuo and the residue was used directly without further purification. To a solution of the above crude product and 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine (96.64 mg, 0.64 mmol, 2.0 equiv) in DMF (5 mL) was added HOBT (64.8 mg, 0.48 mmol, 1.5 equiv), EDCI (104.45 mg, 0.54 mmol, 1.7 equiv) and _Et3N (0.17 mL, 1.28 mmol, 4 equiv). The mixture was stirred at room temperature overnight and diluted with water. The organic layer was separated and the aqueous layer was extracted with DCM. The combined extracts were dried and concentrated. The residue was purified by preparative HPLC to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (30 mg, 18% over 2 steps) as an off-white solid. LRMS (M+H ⁺ ) m/z calcd. 490.2, found 490.0. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ: 8.93 (s, 1H), 8.67 (d, 2H), 8.27 (s, 2H), 8.16 (s, 1H), 7.82 (s, 1H), 7.63 (d, 1H), 6.13 (s, 1H), 5.70 (s, 2H), 4.44 (s, 2H), 4.35 (s, 2H), 3.58 (s, 3H), 2.52 (s, 3H), 2.31 (s, 3H), 2.17 (s, 3H).

Example 16: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (Example 15) was prepared as a yellow solid (60 mg, 35% over 2 steps). LRMS (M+H ⁺ ) m/z calcd 512.2, found 512.0.

¹ H NMR (DMSO-d ₆ , 400MHz) δ8.85 (s, 1H), 8.64 (d, 1H), 8.31 (s, 1H), 8.11 (s, 1H), 8.05 (d, 2H), 7.81 (s, 1H), 7.69 (d , 2H), 7.60(s, 1H), 7.47(d, 1H), 6.91(d, 1H), 4.69(s, 2H), 4.45(s, 2H), 3.52(s, 3H), 2.52(s, 3H).

Example 17: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((3-Chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (Example 15) was prepared as a yellow solid (30 mg, 17% over 2 steps). LRMS (M+H ⁺ ) m/z calcd 537.1, found 537.1.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.40(s,1H),9.26(t,1H),8.93(d,1H),8.67(d,1H),8.28(d,2H),8.16(s,1H),7.86(s,1H), 7.68(d,1H),7.51(d,1H),7.46(d,1H),7.24(d,1H),4.60(d,2H),4.45(s,2H),3.57(s,3H),2.50(s,3H).

Example 18: Preparation of N-((6-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-Fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-Fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (40 mg, 28% over 2 steps) was prepared as an off-white solid as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (Example 15). LRMS (M+H ⁺ ) m/z calcd 503.2, found 503.0. ¹ H NMR (DMSO-d ₆ ,400MHz)δ11.12(s,1H),9.24(t,1H),8.92(d,1H),8.68(d,1H),8.28(d,2H),8.16(s,1H),7.89(s,1H),7.70(d ,1H),7.52(d,1H),7.32(s,1H),7.19(d,1H),6.41(d,1H),4.59(d,2H),4.46(s,1H),3.58(s,3H),2.51(s,3H).

Example 19: Preparation of N-((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((3-Chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (55 mg, 38%) was prepared as a white solid as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (Example 15). LRMS (M+H ⁺ ) m/z calcd 520.1, found 519.8.

¹ H NMR (DMSO-d ₆ , 400MHz) δ9.35 (t, 1H), 8.92 (s, 1H), 8.66 (d, 1H), 8.26-8.32 (m, 3H), 8.16 (s, 1H), 7.88 (s , 1H), 7.85 (s, 1H), 7.65-7.68 (m, 2H), 4.59 (d, 2H), 4.45 (s, 2H), 3.56 (s, 3H), 2.51 (s, 3H).

Example 20: Preparation of N-((5-chloro-1H-indazol-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((5-chloro-1H-indazol-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((5-Chloro-1H-indazol-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (Example 15) was prepared as a yellow solid (30 mg, 21%). LRMS (M+H ⁺ ) m/z calcd 520.1, found 519.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.41(t,1H),8.92(s,1H),8.66(d,1H),8.27(d,2H),8.15(s,1H),7.91(s,1H),7.86(s ,1H),7.65(d,1H),7.53(d,1H),7.33(d,1H),4.79(d,2H),4.45(s,2H),3.56(s,3H),2.51(s,3H).

Example 21: Preparation of N-((3-chloro-4-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-4-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((3-Chloro-4-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (40 mg, 27%) was prepared as a white solid as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (Example 15). LRMS (M+H ⁺ ) m/z calcd 537.1, found 536.7.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.60(s,1H),9.26(t,1H),8.92(s,1H),8.66(d,1H),8.27(d,2H),8.16(s,1H),7.85(s,1 H),7.65(d,1H),7.52(d,1H),7.12-7.21(m,2H),4.58(d,2H),4.45(s,2H),3.57(s,3H),2.51(s,3H).

Example 22: Preparation of N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(5-chloro-1-oxo-1H-isoquinolin-2-ylmethyl)-isonicotinamide

N-(6-Amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(5-chloro-1-oxo-1H-isoquinolin-2-ylmethyl)-isonicotinamide

Step 1: Preparation of methyl 2-chlorophenethylcarbamate

To a solution of 2-(2-chloro-phenyl)-ethylamine (10 g, 64 mmol, 1.0 equiv) in DCM (150 mL) was added TEA (12.9 g, 128 mmol, 2.0 equiv). The mixture was stirred at 0 ° C. and methyl chloroformate (9.07 g, 96 mmol, 1.5 equiv) was added under a nitrogen atmosphere. The mixture was allowed to warm to room temperature and stirred for 1 h, concentrated, and purified by silica gel chromatography (PE/EtOAc=5/1, v/v) to give methyl 2-chlorophenethylcarbamate (9.0 g, 66%) as a colorless oil.

Step 2: Preparation of 5-chloro-3,4-dihydro-2H-isoquinolin-1-one

Trifluoromethanesulfonic acid (170 mL, 2.2 mol, 50 equiv) was added to N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(5-chloro-1-oxo-1H-isoquinolin-2-ylmethyl)-isonicotinamide (9.0 g, 44.2 mmol, 1.0 equiv) at 0° C. The mixture was stirred at 70° C. under nitrogen for 24 h. The mixture was then poured into ice water to give 5-chloro-3,4-dihydro-2H-isoquinolin-1-one (5.1 g, 67%) as a yellow oil.

Step 3: Preparation of 5-chloro-2H-isoquinolin-1-one

To a solution of 5-chloro-3,4-dihydro-2H-isoquinolin-1-one (5.1 g, 28 mmol, 1.0 equiv) in dioxane (150 mL) was added DDQ (22 g, 70 mmol, 3.4 equiv). The mixture was stirred at 100 ° C for 72 h. The solvent was removed and EtOAc was added. The organic layer was then washed with 10% NaOH, concentrated, and purified by silica gel chromatography (PE/EtOAc=3/1, v/v) to give 5-chloro-2H-isoquinolin-1-one (1.3 g, 25%) as an orange oil.

Step 4: Preparation of methyl 2-((5-chloro-1-oxoisoquinolin-2(1H)-yl)methyl)isonicotinate

To a solution of 5-chloro-2H-isoquinolin-1-one (470 mg, 2.6 mmol, 1.0 eq) in DMF (10 mL) was added K ₂ CO ₃ (723 mg, 5.2 mmol, 2.0 eq) and methyl 2-(chloromethyl)isonicotinate (722 mg, 3.9 mmol, 1.5 eq). The mixture was stirred at 30° C. for 4 h. The solvent was removed and the mixture was purified by silica gel chromatography (PE/EtOAc=3/1, v/v) to give methyl 2-((5-chloro-1-oxoisoquinolin-2(1H)-yl)methyl)isonicotinate (710 mg, 83%) as a yellow solid.

Step 5: Preparation of N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(5-chloro-1-oxo-1H-isoquinolin-2-ylmethyl)-isonicotinamide

To a solution of methyl 2-((5-chloro-1-oxoisoquinolin-2(1H)-yl)methyl)isonicotinate (210 mg, 0.54 mmol, 1.0 equiv) in THF (5 mL)/H ₂ O (5 mL) was added NaOH (43 mg, 1.08 mmol, 2.0 equiv). The mixture was stirred at 40° C. for 1 h and acidified to pH 5 with 1N HCl solution. The mixture was concentrated in vacuo and the residue was used without further purification. To a solution of the above crude product and 4-aminomethyl-3,5-dimethyl-aniline (122 mg, 0.81 mmol, 1.5 equiv) in DMF (8 mL) was added HATU (230 mg, 0.6 mmol, 1.2 equiv) and Et ₃ N (0.3 mL, 1.62 mmol, 3 equiv). The mixture was stirred at room temperature overnight, concentrated, and purified by flash chromatography on a silica gel column (DCM/MeOH=10/1, v/v) to give N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(5-chloro-1-oxo-1H-isoquinolin-2-ylmethyl)-isonicotinamide (31 mg, 13% over 2 steps) as a white solid. LRMS (M+H ⁺ ) m/z calcd 448.2, found 448.0. ¹ HNMR(DMSO-d ₆ ,400MHz)δ8.71(m,1H),8.56-8.57(d,1H),8.16-8.18(d,1H),7.88-7.90(d,1H),7.74-7.76(d,1H),7.70(s,1H),7.64-7.65(d, 1H),7.48-7.52(t,1H),6.81-6.83(d,1H),6.14(s,1H),5.80(m,2H),5.34(s,2H),4.33-4.34(d,2H),2.30(s,3H),2.17(s,3H).

Example 23: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((6-chloro-1-oxoisoquinolin-2(1H)-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((6-chloro-1-oxoisoquinolin-2(1H)-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-ylmethyl)methyl)-2-((6-chloro-1-oxoisoquinolin-2(1H)-yl)methyl)isonicotinamide (45 mg, 33%) was prepared as a white solid as described for N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(5-chloro-1-oxo-1H-isoquinolin-2-ylmethyl)-isonicotinamide (Example 22). LRMS (M+H ⁺ ) m/z calcd 448.2, found 447.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ8.71(s,1H),8.58(d,1H),8.17(d,1H),7.83(s,1H),7.68-7.65(m,3H),7.54( dd,1H),6.67(d,1H),6.15(s,1H),5.74(br,2H),4.35(d,2H),2.31(s,3H),2.17(s,3H).

Example 24: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-1H-indol-5-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-1H-indol-5-yl)methyl)isonicotinamide

Step 1: Preparation of methyl 3-chloro-1H-indole-5-carboxylate

To the MeOH solution of 1H-indole-5-methyl-formiate (10.0g, 57.1mmol, 1.0 equivalent) was added NCS (8.4g, 62.8mmol, 1.1 equivalent). The mixture was stirred at room temperature for 3h. MeOH was removed by evaporation, and the residue was redissolved in EtOAc. The mixture was washed twice with salt water. The organic layer was dried and concentrated to obtain 3-chloro-1H-indole-5-methyl-formiate (quantitative) as a yellow solid.

Step 2: Preparation of 1-tert-butyl 5-methyl 3-chloro-1H-indole-1,5-dicarboxylate

To a solution of methyl 3-chloro-1H-indole-5-carboxylate (11.9 g, 57.1 mmol, 1.0 equiv) in MeOH was added Boc ₂ O (18.7 g, 86.7 mmol, 1.5 equiv) and DMAP (348 mg, 2.86 mmol, 0.05 equiv). The mixture was stirred at room temperature for 2 h. The mixture was concentrated and purified by chromatography on a silica gel column (EtOAc/PE=1/10, v/v) to give 1-tert-butyl 5-methyl 3-chloro-1H-indole-1,5-dicarboxylate (13.4 g, 76%) as an off-white solid.

Step 3: Preparation of tert-butyl 3-chloro-5-(hydroxymethyl)-1H-indole-1-carboxylate

To a THF (100 mL) solution of 1-tert-butyl 5-methyl 3-chloro-1H-indole-1,5-dicarboxylate (7.0 g, 22.6 mmol, 1 equivalent) was added LiAlH(t-BuO) ₃ (14.4 g, 56.6 mmol, 2.5 equivalents). The resulting mixture was stirred at 60 ° C for 12 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=2/1, v/v) to give tert-butyl 3-chloro-5-(hydroxymethyl)-1H-indole-1-carboxylate (4.3 g, 68%) as a white solid.

Step 4: Preparation of tert-butyl 3-chloro-5-(chloromethyl)-1H-indole-1-carboxylate

To a solution of tert-butyl 3-chloro-5-(hydroxymethyl)-1H-indole-1-carboxylate (1.5 g, 5.34 mmol, 1 equiv) in anhydrous DCM (30 mL) was added _Et3N (1.5 mL, 10.68 mmol, 2 equiv) and MsCl (0.62 mL, 8.01 mmol, 1.5 equiv). The resulting mixture was stirred at room temperature for 24 h and then quenched by adding water. The mixture was extracted with DCM. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=20/1, v/v) to give tert-butyl 3-chloro-5-(chloromethyl)-1H-indole-1-carboxylate (1.17 g, 73%) as a white solid.

Step 5: Preparation of tert-butyl 3-chloro-5-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-1H-indole-1-carboxylate

To a solution of tert-butyl 3-chloro-5-(chloromethyl)-1H-indole-1-carboxylate (1.1 g, 3.68 mmol, 1.0 equiv) in dioxane (20 mL) was added methyl 2-(trimethylstannyl)isonicotinate (1.22 g, 4.05 mmol, 1.1 equiv) and Pd( _PPh3 ) _{2Cl2 (} 260 mg, 0.37 mmol, 0.1 equiv). The mixture was stirred at 90°C for 3 h under a nitrogen atmosphere, concentrated, and purified by silica gel chromatography (PE/EtOAc=20/1, v/v) to give tert-butyl 3-chloro-5-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-1H-indole-1-carboxylate (690 mg, 47%) as an off-white solid.

Step 6: Preparation of 2-(3-chloro-1H-indol-5-ylmethyl)-isonicotinic acid

To a solution of tert-butyl 3-chloro-5-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-1H-indole-1-carboxylate (690 mg, 1.72 mmol, 1.0 equivalent) in DCM (3 mL) was added TFA (5 mL). The mixture was stirred at room temperature for 2 h. The mixture was then concentrated, and the residue was redissolved in DCM and washed with saturated NaHCO ₃ aqueous solution. The organic layer was concentrated. The residue was dissolved in THF/H ₂ O (5 mL, v/v=1: 1). NaOH was added to the mixture. The mixture was stirred at room temperature for 0.5 h. The mixture was then acidified to pH 5 with 1N HCl. The mixture was extracted with EtOAc. The combined organic layers were dried and concentrated to give 2-(3-chloro-1H-indol-5-ylmethyl)-isonicotinic acid (220 mg, 45%) as a yellow solid.

Step 7: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-1H-indol-5-yl)methyl)isonicotinamide

To a solution of 2-(3-chloro-1H-indol-5-ylmethyl)-isonicotinic acid (100 mg, 0.35 mmol, 1 eq) and 4-aminomethyl-3,5-dimethyl-aniline (117 mg, 0.52 mmol, 1.5 eq) in DMF (5 mL) was added HATU (160 mg, 0.42 mmol, 1.2 eq) and _Et3N (140 mg, 1.40 mmol, 4 eq). The mixture was stirred at room temperature for 1 h, concentrated, and purified by preparative HPLC to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-1H-indol-5-yl)methyl)isonicotinamide (35 mg, 24%) as an off-white solid. LRMS (M+H ⁺ ) m/z calcd 420.2, found 419.8.

¹ H NMR (CD ₃ OD,400MHz)δ8.54(d,1H),7.61(s,1H),7.54(dd,1H),7.38(s,1H),7.29(d,1H),7.20( s,1H),7.06(dd,1H),6.28(s,1H),4.45(s,2H),4.25(d,2H),2.36(s,3H),2.22(s,3H).

Example 25: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-1H-indol-5-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-1H-indol-5-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-1H-indol-5-yl)methyl)isonicotinamide (25 mg, 16%) was prepared as a white solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 400.2, found 399.9. ¹ H NMR (CD ₃ OD,400MHz)δ8.53(d,1H),7.59(s,1H),7.52(d,1H),7.37(s,1H),7.23(d,1H),6.97(d,1H), 6.95(s,1H),6.27(s,1H),4.45(s,2H),4.24(d,2H),2.35(s,3H),2.25(s,3H),2.22(s,3H).

Example 26: Preparation of N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of 5-aminomethyl-6-methyl-pyridin-2-ylamine hydrochloride

A mixture of 6-amino-2-methyl-nicotinonitrile (2 g, 15.0 mmol, 1 equiv), Pd/C (10%, 500 mg) and concentrated HCl (3 mL) in EtOH/MeOH (10 mL/10 mL) was stirred at room temperature overnight under H ₂ (50 psi). The reaction mixture was filtered and the filtrate was concentrated to give crude 5-aminomethyl-6-methyl-pyridin-2-ylamine hydrochloride (3.5 g) as a yellow solid. LRMS (M+H ⁺ ) m/z calcd 138, found 138.

Step 2: Preparation of tert-butyl (6-amino-2-methylpyridin-3-yl)methylcarbamate

To a mixture of 5-aminomethyl-6-methyl-pyridin-2-ylamine hydrochloride (3.5 g, crude product) in DCM (50 mL) was added TEA (4.5 g, 45.0 mmol, 3 equiv.) followed by Boc ₂ O (4.9 g, 22.5 mmol, 1.5 equiv.). The reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=20/1, v/v) to give tert-butyl (6-amino-2-methylpyridin-3-yl)methylcarbamate (3 g) as a white solid. LRMS (M+H ⁺ ) m/z calculated value 238, found 238.

Step 3: Preparation of tert-butyl (6-amino-5-bromo-2-methylpyridin-3-yl)methylcarbamate

To a solution of tert-butyl (6-amino-2-methylpyridin-3-yl)methylcarbamate (3 g, 13.7 mmol, 1 equiv) in DCM (30 mL) was added NBS (2.5 g, 13.9 mmol, 1.1 equiv). The resulting mixture was concentrated. The residue was purified by column chromatography on silica gel (PE/EtOAc=10/1 to EtOAc, v/v) to give tert-butyl (6-amino-5-bromo-2-methylpyridin-3-yl)methylcarbamate (2 g, 50%) as a white solid. LRMS (M+H ⁺ ) m/z calculated value 316,318, found value 316,318.

Step 4: Preparation of tert-butyl (6-amino-5-cyano-2-methylpyridin-3-yl)methylcarbamate

A mixture of tert-butyl (6-amino-5-bromo-2-methylpyridin-3-yl)methylcarbamate (500 mg, 1.58 mmol, 1 eq), Zn(CN) ₍ 185 mg, 1.58 mmol, 1.0 eq) and Pd( _PPh ) ₍ 182 mg, 0.16 mmol, 0.1 eq) in DMF (20 mL) was heated to 95 ° C and stirred for 3 h. It was then cooled to room temperature, filtered, and the filtrate was concentrated. The residue was purified by column chromatography on silica gel (PE/EtOAc=2/1, v/v) to give tert-butyl (6-amino-5-cyano-2-methylpyridin-3-yl)methylcarbamate (300 mg, 72%) as a white solid.

LRMS (M+H ⁺ ) m/z calcd. 263, found 263.

Step 5: Preparation of 2-amino-5-aminomethyl-6-methyl-nicotinonitrile hydrochloride

To a mixture of tert-butyl (6-amino-5-cyano-2-methylpyridin-3-yl)methylcarbamate (1.2 g, 4.6 mmol, 1 equivalent) in EtOAc (10 mL) was added 10 mL of 6N HCl in EtOAc and stirred for 2 h. The reaction mixture was filtered and the filter cake was washed with EtOAc to afford 2-amino-5-aminomethyl-6-methyl-nicotinonitrile hydrochloride (600 mg, 67%) as a white solid. LRMS (M+H ⁺ ) m/z calculated value 163, found value 163.

Step 6: Preparation of N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(3-chloro-quinolin-6-ylmethyl)-isonicotinic acid (80 mg, 0.27 mmol, 1 eq) in DMF (10 mL) was added 2-amino-5-aminomethyl-6-methyl-nicotinonitrile hydrochloride (54 mg, 0.27 mmol, 1 eq), followed by EDCI (78 mg, 0.41 mmol, 1.5 eq), HOBT (55 mg, 0.41 mmol, 1.5 eq), and TEA (82 mg, 0.81 mmol, 3.0 eq). The reaction mixture was heated to 45° C. and stirred overnight. Water was added, and the mixture was extracted with DCM. The organic layer was washed with water, dried over Na ₂ SO ₄ , filtered, and concentrated. The residue was purified by preparative HPLC to give N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (60 mg, 50%) as a white solid. LRMS (M+H ⁺ ) m/z calcd 443.1, found 442.8.

¹ H NMR (DMSO-d ₆ ,400MHz): δ9.04(t,1H),8.83(d,1H),8.64(d,1H),8.53(s,1H),7.98(d,1H),7.85(s,1H),7.77(s ,1H),7.74(d,1H),7.68(s,1H),7.63(s,1H),6.78(s,2H),4.37(s,2H),4.31(d,2H),2.36(s,3H).

Example 27: Preparation of 2-amino-5-((2-((3-chloroquinolin-6-yl)methyl)isonicotinamido)methyl)-6-methylnicotinamide

2-Amino-5-((2-((3-chloroquinolin-6-yl)methyl)isonicotinamido)methyl)-6-methylnicotinamide

[0266] To a solution of N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (80 mg, 0.18 mmol, 1 eq) in DMF (5 mL) was added _{KCO (} 50 mg, 0.36 mmol, 2.0 eq) followed by ₃₀ % _HO (2 mL). The reaction mixture was heated to 50°C with stirring for 3 h. The reaction mixture was concentrated. The residue was purified by preparative HPLC to give 2-amino-5-((2-((3-chloroquinolin-6-yl)methyl)isonicotinamido)methyl)-6-methylnicotinamide (45 mg, 54%) as a white solid. LRMS (M+H ⁺ ) m/z calcd 461.1, found 461.1.

¹ H NMR (DMSO-d ₆ , 400MHz) δ8.91(t,1H),8.83(d,1H),8.63(d,1H),8.53(d,1H),7.98(d,1H),7.86(s,3H),7.7 8(s,1H),7.74(d,1H),7.64(d,1H),7.13-7.21(m,3H),4.36(s,2H),4.32(d,2H),2.31(s,3H).

Example 28: Preparation of N-((6-amino-5-chloro-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-5-chloro-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of 6-amino-5-chloro-2-methyl-nicotinonitrile

A mixture of 6-amino-2-methyl-nicotinonitrile (500 mg, 3.76 mmol, 1 equivalent) and NCS (1 g, 7.52 mmol, 2 equivalents) in AcOH (10 mL) was heated to 60 ° C and kept stirring for 2 h. The reaction mixture was concentrated. The residue was purified by column chromatography on silica gel (PE/EtOAc=10/1 to EtOAc v/v) to give 6-amino-5-chloro-2-methyl-nicotinonitrile (400 mg, 64%) as a white solid. LRMS (M+H ⁺ ) m/z calculated value 168, found value 168.

Step 2: Preparation of 5-aminomethyl-3-chloro-6-methyl-pyridin-2-ylamine hydrochloride

A mixture of 6-amino-5-chloro-2-methyl-nicotinonitrile (400 mg, 2.4 mmol, 1 equiv), Raney nickel (400 mg) and concentrated HCl (1 mL) in EtOH/MeOH (10 mL/10 mL) was stirred overnight at room temperature under H ₂ (1 atm). The reaction mixture was filtered and the filtrate was concentrated to give crude 5-aminomethyl-3-chloro-6-methyl-pyridin-2-ylamine hydrochloride (1 g) as a yellow solid. LRMS (M+H ⁺ ) m/z calculated value 172, found value 172.

Step 3: Preparation of N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-Amino-5-chloro-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (15 mg, 12%) was prepared as a white solid as described for N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (Example 26). LRMS (M+H ⁺ ) m/z calcd 452.1, found 451.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.03(t,1H),8.83(d,1H),8.63(d,1H),8.52(d,1H),7.98(d,1H),7.85(s,1H),7.7 2-7.76(m,2H),7.62(d,1H),7.41(s,1H),6.11(s,2H),4.36(s,2H),4.29(d,2H),2.29(s,3H).

Example 29: Preparation of N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (25 mg, 22%) was prepared as a white solid as described for N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (Example 26). LRMS (M+H ⁺ ) m/z calcd 418.1, found 417.8.

¹ H NMR (DMSO-d ₆ , 300MHz) δ8.93 (t, 1H), 8.83 (d, 1H), 8.62 (d, 1H), 8.52 (d, 1H), 7.98 (d, 1H), 7.84 (s, 1H), 7.7 1-7.77(m, 2H), 7.61(d, 1H), 6.24(s, 1H), 5.75(s, 2H), 4.37(s, 2H), 4.29(d, 2H), 2.14(s, 3H).

Example 30: Preparation of N-((6-amino-2-(trifluoromethyl)pyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-(trifluoromethyl)pyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-(trifluoromethyl)pyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (45 mg, 35%) was prepared as a white solid as described for N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (Example 26). LRMS (M+H ⁺ ) m/z calcd 472.1, found 471.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.13(t,1H),8.83(d,1H),8.65(d,1H),8.53(d,1H),7.98(d,1H),7.86(s,1H),7.77(s ,1H),7.74(d,1H),7.64(d,1H),7.52(d,1H),6.65(d,1H),6.44(s,2H),4.44(d,2H),4.37(s,2H).

Example 31: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (65 mg, 45%) was prepared as a white solid as described for N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (Example 26). LRMS (M+H ⁺ ) m/z calcd 454.1, found 453.9.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.40(t,1H),8.83(d,1H),8.67(d,1H),8.54(s,1H),8.13(d,1H),7.97(d,1H),7.87(s,1H),7.81(s,1H ),7.74-7.76(m,2H),7.69(d,1H),7.55(s,1H),7.40(d,1H),6.84(d,1H),6.74(s,2H),4.61(d,2H),4.38(s,2H).

Example 32: Preparation of 2-(3-chloro-quinolin-6-ylmethyl)-N-(6-fluoro-1H-indazol-5-ylmethyl)-isonicotinamide

2-(3-Chloro-quinolin-6-ylmethyl)-N-(6-fluoro-1H-indazol-5-ylmethyl)-isonicotinamide

Step 1: Preparation of 4-amino-2-fluoro-5-methyl-benzonitrile

To a solution of 4-bromo-5-fluoro-2-methyl-aniline (20 g, 98.0 mmol, 1.0 _equiv ) in DMF (100 mL) was added Zn(CN) (28.7 g, 245 mmol, 2.5 equiv) followed by the addition of Pd( _PPh ) _{under N.} The mixture was stirred at 90 ° C overnight. The mixture was concentrated in vacuo and the residue was dissolved in water and extracted with EtOAc. The combined extracts were dried and concentrated. The residue was purified by column chromatography on silica gel (PE/EtOAc=10/1, v/v) to give 4-amino-2-fluoro-5-methyl-benzonitrile (13.68 g, 92%) as a yellow solid.

Step 2: Preparation of 6-fluoro-1H-indazole-5-carbonitrile

To a solution of 4-amino-2-fluoro-5-methyl-benzonitrile (13.68 g, 90.59 mmol, 1.0 equiv) in AcOH (450 mL) was added NaNO ₂ (7.5 g, 108.7 mmol, 1.2 equiv). The mixture was stirred at room temperature overnight. After completion, an aqueous NaOH solution (50%) was added to the reaction mixture until the pH was 7-8. The mixture was extracted with EtOAc. The organic layer was concentrated under pressure. The residue was purified by column chromatography on a silica gel column (PE/EtOAc=15/1, v/v) to give 6-fluoro-1H-indazole-5-carbonitrile (5 g, 34%) as a white solid.

Step 3: Preparation of 6-fluoro-1-(tetrahydro-pyran-2-yl)-1H-indazole-5-carbonitrile

To a solution of 6-fluoro-1H-indazole-5-carbonitrile (5 g, 31.05 mmol, 1.0 equiv) and 3,4-dihydro-2H-pyran (5.25 g, 62.1 mmol, 2 equiv) in DCM (50 mL) was added PTSA (590 mg, 3.11 mmol, 0.1 equiv), and the mixture was stirred at room temperature overnight. The solvent was removed in vacuo. The residue was dissolved in EtOAc, washed with water, brine, and dried over Na ₂ SO _4. The combined extracts were dried and concentrated. The residue was purified by chromatography on a silica gel column (PE/EtOAc=15/1, v/v) to give 6-fluoro-1-(tetrahydro-pyran-2-yl)-1H-indazole-5-carbonitrile (4.39 g, 57%) as a brown solid.

Step 4: Preparation of (6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-5-yl)methanamine

Under H ₂ , to a MeOH (20 mL) solution of 6-fluoro-1-(tetrahydro-pyrans-2-yl)-1H-indazole-5-carbonitrile (4.39 g, 17.92 mmol, 1.0 equivalent) was added Raney nickel (800 mg). The mixture was stirred at room temperature overnight. The reaction mixture was filtered, and the filtrate was purified by chromatography on a silica gel column (PE/EtOAc=15/1, v/v) to obtain (6-fluoro-1-(tetrahydro-2H-pyrans-2-yl)-1H-indazole-5-yl)methylamine (3.8 g, 85%) as a white solid.

Step 5: Preparation of (6-fluoro-1H-indazol-5-yl)methylamine hydrochloride

To a solution of (6-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-5-yl)methanamine (3.43 g, 15.26 mmol, 1 equivalent) in EtOAc was added EtOAc/HCl (10 M). The mixture was stirred at room temperature for 3 h. The reaction mixture was filtered and the filtrate was concentrated to give (6-fluoro-1H-indazol-5-yl)methanamine hydrochloride (3.43 mg, crude).

Step 6: Preparation of 2-(3-chloro-quinolin-6-ylmethyl)-N-(6-fluoro-1H-indazol-5-ylmethyl)-isonicotinamide

To a solution of (6-fluoro-1H-indazole-5-yl)-methylamine hydrochloride (80 mg, 0.4 mmol, 1.5 equiv) in DMF (10 mL) was added 2-(3-chloro-quinolin-6-ylmethyl)-isonicotinic acid (80 mg, 0.26 mmol, 1 equiv), HATU (122 mg, 0.32 mmol/1.2 equiv) and TEA (1 mL). The reaction mixture was stirred at room temperature for 3 h. It was then quenched with water and extracted with DCM. The combined extracts were dried and concentrated, and the residue was purified by preparative HPLC to give 2-(3-chloro-quinolin-6-ylmethyl)-N-(6-fluoro-1H-indazole-5-ylmethyl)-isonicotinamide (30 mg, 27%) as a white solid. LCMS (M+H ⁺ ) m/z calculated value 446.1, found value 446.0.

^1H NMR(DMSO-d6,400MHz)δ13.09(s,1H),9.26-9.28(t,1H),8.83-8.84(d,1H),8.65-8.66(d,1H),8.52-8.53(d,1H),8.06(s,1H),7.97-7.9 9(d,1H),7.86(s,1H),7.80(s,1H),7.68-7.76(m,2H),7.67-7.68(d, 1H),7.32-7.35(d,1H),4.56-4.57(d,2H),4.38(s,2H),2.50(s,3H).

Example 33: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((3-Chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (50 mg, 33%) was prepared as a yellow solid as described for N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (Example 26). LRMS (M+H ⁺ ) m/z calcd 479.1, found 478.9.

¹ H NMR (DMSO-d ₆ ,400MHz): δ11.42(br,1H),9.25(t,1H),8.83(s,1H),8.65(d,1H),8.53(s,1H),7.98(d,1H),7.85(s,1H) ,7.79(s,1H),7.74(d,1H),7.66(d,1H),7.51(s,1H),7.44(d,1H),7.22(d,1H),4.58(d,2H),4.37(s,2H).

Example 34: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (Example 26) was prepared as a yellow solid (60 mg, 45%). LRMS (M+H ⁺ ) m/z calcd 418.1, found 417.8

¹ H NMR (DMSO-d ₆ , 400MHz) δ9.00 (t, 1H), 8.83 (d, 1H), 8.63 (d, 1H), 8.53 (d, 1H), 7.98 (d, 1H), 7.85 (s, 1H), 7.73-7.76 (m, 2H), 7.62 (d, 1H), 7.77 (d, 1H), 6.26 (d, 1H), 5.85 (s, 2H), 4.36 (s, 2H), 4.28 (d, 2H), 2.28 (s, 3H).

Example 35: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (40 mg, 29%) was prepared as a yellow solid as described for N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (Example 26). LRMS (M+H ⁺ ) m/z calcd 432.2 found 432.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ8.83(d,1H),8.64(t,1H),8.60(d,1H),8.53(s,1H),7.97(d,1H),7.84(s,2H),7.72-7 .75(m,2H),7.60(d,1H),6.11(s,1H),5.67(s,2H),4.33-4.35(m,4H),2.29(s,3H),2.15(s,3H).

Example 36: Preparation of N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((3-Aminobenzo[d]isoxazol-6-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (65 mg, 46%) was prepared as a yellow solid as described for N-((6-amino-5-cyano-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (Example 26). LRMS (M+H ⁺ ) m/z calcd 444.1 found 443.9.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.38(t,1H),8.83(d,1H),8.67(s,1H),8.53(d,1H),7.99(d,1H),7.86(s,1H),7.81(s ,1H),7.76(d,2H),7.67(d,1H),7.36(s,1H),7.21(d,1H),6.38(s,2H),4.60(d,2H),4.38(s,2H).

Example 37: Preparation of N-(5-chloro-1H-indazol-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

N-(5-Chloro-1H-indazol-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

To a solution of methyl 2-((3-chloroquinolin-6-yl)methyl)isonicotinate (1.9 g, 6.1 mmol, 1.0 equiv) in THF (15 mL)/H ₂ O (15 mL) was added NaOH (360 mg, 9.11 mmol, 1.5 equiv). The mixture was stirred at 40° C. for 1 h and acidified to pH 5 with 1N HCl solution to afford 0.8 g of the product. To a solution of the above crude product (100 mg, 0.33 mmol, 1.0 equiv) and (5-chloro-1H-indazol-3-yl)-methylamine (120 mg, 0.66 mmol, 2.0 equiv) in DMF (8 mL) was added HATU (152 mg, 0.4 mmol, 1.2 equiv) and Et ₃ N (0.15 mL, 0.99 mmol, 3 equiv). The mixture was stirred at room temperature overnight, concentrated, and purified by flash chromatography on a silica gel column (DCM/MeOH=10/1, v/v) to give N-(5-chloro-1H-indazol-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide (46 mg, 30% over 2 steps) as a white solid. LRMS (M+H ⁺ ) m/z calcd 462.1, found 461.9.

¹ H NMR (DMSO-d ₆ ,400MHz) δ13.09 (s, 1H), 9.40 (m, 1H), 8.82-8.83 (d, 1H), 8.63-8.64 (d, 1H), 8.51-8.52 (d, 1H), 7.9 6-7.98(d,1H),7.89(s,1H),7.84(s,1H),7.78(s,1H),7.74(d,1H),7.64(d,1H),7.52-7.54(d,1H),

7.32-7.34 (dd, 1H), 4.77-4.78 (d, 2H), 4.36 (s, 2H).

Example 38: Preparation of N-(3-chloro-1H-pyrrolo[2,3-b]pyridin-5-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

N-(3-Chloro-1H-pyrrolo[2,3-b]pyridin-5-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

To a solution of 2-(3-chloro-quinolin-6-ylmethyl)-isonicotinic acid (100 mg, 0.33 mmol, 1.0 equiv) and (3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)-methylamine (120 mg, 0.66 mmol, 2.0 equiv) in DMF (8 mL) was added HATU (152 mg, 0.4 mmol, 1.2 equiv) and _Et3N (0.15 mL, 0.99 mmol, 3 equiv). The mixture was stirred at room temperature overnight, concentrated, and purified by flash chromatography on a silica gel column (DCM/MeOH=10/1, v/v) to give N-(5-chloro-1H-indazol-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide (13 mg, 8.5%) as a white solid. LRMS (M+H ⁺ ) m/z calcd. 462.1, found 461.9.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.96(s,1H),9.33(m,1H),8.82(d,1H),8.65(d,1H),8.51-8.52(d,1H),8.3 0(d,1H),7.98(d,1H),7.86(m,2H),7.63-7.77(m,4H),4.57-4.59(d,2H),4.36(s,2H).

Example 39: Preparation of N-(6-amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

N-(6-Amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

N-(6-Amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide (41 mg, 30%) was prepared as a white solid as described for N-(5-chloro-1H-indazol-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide (Example 37). LRMS (M+H ⁺ ) m/z calcd 418.1, found 418.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ8.99(m,1H),8.83(d,1H),8.62-8.63(d,1H),8.53(d,1H),7.96-7.99(d,1H),7.85(s,1H),7.7 2-7.76(m,2H),7.61-7.63(d,1H),6.21-6.23(d,2H),5.75(s,2H),4.36(s,2H),4.27(d,2H),2.27(s,2H).

Example 40: Preparation of N-(3-chloro-4-fluoro-1H-indol-5-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

N-(3-Chloro-4-fluoro-1H-indol-5-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

N-(3-Chloro-4-fluoro-1H-indol-5-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide (41 mg, 26%) was prepared as a white solid as described for N-(5-chloro-1H-indazol-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide (Example 37). LRMS (M+H ⁺ ) m/z calcd 479.1, found 479.0.

¹ H NMR (DMSO-d ₆ , 400MHz) δ11.59 (s, 1H), 9.23 (m, 1H), 8.82 (d, 1H), 8.63 (d, 1H), 8.52 (d, 1H), 7.98 (d, 1H), 7.84 (s, 1H), 7.78 (s, 1H), 7.72-7.74(d, 1H), 7.63-7.64(d, 1H), 7.50-7.51(d, 1H), 7.12-7.19(m, 2H), 4.55-4.57(d, 2H), 4.36(s, 2H).

Example 41: Preparation of 2-(3-chloro-quinolin-6-ylmethyl)-N-(6-fluoro-1H-indol-5-ylmethyl)-isonicotinamide

2-(3-Chloro-quinolin-6-ylmethyl)-N-(6-fluoro-1H-indol-5-ylmethyl)-isonicotinamide

2-(3-Chloro-quinolin-6-ylmethyl)-N-(6-fluoro-1H-indol-5-ylmethyl)-isonicotinamide (71 mg, 48%) was prepared as a white solid as described for N-(5-chloro-1H-indazol-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide (Example 37). LRMS (M+H ⁺ ) m/z calcd 445.1, found 445.0.

¹ H NMR (DMSO-d ₆ , 400 MHz) δ 11.11 (s, 1H), 9.21 (m, 1H), 8.82 (d, 1H), 8.63 (d, 1H), 8.53 (d, 1H), 7.96 (d, 1H), 7.66-7.85 (m, 4H), 7.47 (d, 1H), 7.17-7.31 (m, 1H), 7.13 (d, 1H), 6.38 (d, 2H), 4.56 (d, 2H), 4.36 (s, 2H). Example 42: Preparation of 2-((3-chloroquinolin-6-yl)methyl)-N-((2-methyl-6-(methylamino)pyridin-3-yl)methyl)isonicotinamide

2-((3-chloroquinolin-6-yl)methyl)-N-((2-methyl-6-(methylamino)pyridin-3-yl)methyl)isonicotinamide

To a solution of sodium methoxide (33 mg, 0.6 mmol, 5.0 equiv) in MeOH (20 mL) was added paraformaldehyde (36 mg, 1.2 mmol, 10.0 equiv) and N-(6-amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide (50 mg, 0.12 mmol, 1.0 equiv). The mixture was stirred at room temperature for 24 h until TLC indicated the starting material was consumed. Sodium borohydride (14 mg, 0.36 mmol, 3.0 equiv) was then added, and the mixture was stirred at 40 ° C for another 3 h. The resulting mixture was concentrated and dissolved in EtOAc, and the organic phase was washed with water and brine, dried, and concentrated in vacuo. The residue was purified by preparative HPLC to give 2-((3-chloroquinolin-6-yl)methyl)-N-((2-methyl-6-(methylamino)pyridin-3-yl)methyl)isonicotinamide (13 mg, 25%). LRMS (M+H ⁺ ) m/z calcd. 432.2, found 431.8.

¹ H NMR(DMSO-d6,400MHz)δ8.98(t,1H),8.82(d,1H),8.63(d,1H),8.52(d,1H),7.98(d,1H),7.85(s,1H),7.74(d,1H),7 .72(d,1H),7.62(d,1H),7.26(d,1H),6.23(d,1H),6.19(d,1H),4.36(s,2H),4.29(d,2H),2.72(d,3H),2.31(s,3H).

Example 43: Preparation of N-((6-amino-2-cyclopropylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-cyclopropylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of 6-amino-2-chloro-nicotinonitrile

To a solution of 6-chloro-5-iodo-pyridin-2-ylamine (25.0 g, 98 mmol, 1.0 equiv) in DMF (200 mL) was added Zn(CN) ₍ 5.7 g, 49 mmol, 0.5 equiv) and _Pd ( _PPh ) (5.66 g, 4.9 mmol, 0.05 equiv). The mixture was stirred overnight at 65° C. under _N. EtOAc and water were then added. The organic layer was concentrated and purified by silica gel chromatography (EA/PE=1/1, v/v) to give 6-amino-2-chloro-nicotinonitrile (12.2 g, 81%) as a yellow solid.

Step 2: Preparation of 6-amino-2-cyclopropyl-nicotinonitrile

To a mixture of 6-amino-2-chloro-nicotinonitrile (3.0 g, 19.6 mmol, 1 eq), cyclopropylboronic acid (2.2 g, 25.5 mmol, 1.3 eq), K ₃ PO ₄ (12.4 g, 58.8 mmol, 3 eq), and tricyclohexylphosphine (550 mg, 1.96 mmol, 0.1 eq) in 200 mL of toluene and 10 mL of water was added Pd(OAc) ₂ (220 mg, 0.98 mmol, 0.05 eq). The reaction mixture was stirred at reflux for 48 h. After cooling to room temperature, the solvent was removed by evaporation. The residue was diluted with water and extracted with EtOAc. The combined organic layers were dried and concentrated. The residue was purified by chromatography on a silica gel column (EtOAc/PE = 1/1, v/v) to give 6-amino-2-cyclopropyl-nicotinonitrile (1.6 g, 51%) as a yellow solid.

Step 3: Preparation of 5-(aminomethyl)-6-cyclopropylpyridin-2-amine hydrochloride

To a solution of 6-amino-2-cyclopropyl-nicotinonitrile (700 mg, 4.4 mmol, 1 eq) was added MeOH (10 mL) and EtOH (10 mL), followed by concentrated HCl. Pd/C was then added under _N2 , and stirring was continued at 40°C overnight. After filtration and washing with MeOH, the organic phase was concentrated under reduced pressure to give the crude product (500 mg, 69%), which was used directly in the next reaction without further purification.

Step 4: Preparation of tert-butyl (6-amino-2-cyclopropylpyridin-3-yl)methylcarbamate

To a solution of 5-aminomethyl-6-cyclopropyl-pyridin-2-ylamine (500 mg, 3.06 mmol, 1 eq) and Boc ₂ O (920 mg, 3.68 mmol, 1.2 eq) in DCM was added TEA (1 mL) and stirred at room temperature for 2 h. The mixture was then washed with water and extracted with EtOAc. After concentration under reduced pressure, the residue was purified by chromatography on a silica gel column (EtOAc/PE=1/2, v/v) to give the title compound (300 mg, 37%).

Step 5: Preparation of 5-(aminomethyl)-6-cyclopropylpyridin-2-amine hydrochloride

To a solution of tert-butyl (6-amino-2-cyclopropylpyridin-3-yl)methylcarbamate in EtOAc was added EtOAc/HCl and stirred at room temperature for 2 h. After filtration and washing with EtOAc, the product was obtained as a white solid (120 mg, 53%), which was used without purification.

Step 6: Preparation of N-((6-amino-2-cyclopropylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

To a solution of 5-aminomethyl-6-cyclopropyl-pyridin-2-ylamine (46 mg, 0.2 mmol, 1.5 equiv) in DMF (10 mL) was added 2-(3-chloro-quinolin-6-ylmethyl)-isonicotinic acid (40 mg, 0.13 mmol, 1 equiv), HOBT (22 mg, 0.16 mmol, 1.2 equiv) and EDCI (30 mg, 0.16 mmol, 1.2 equiv). The reaction mixture was stirred at room temperature overnight. It was then quenched with water and extracted with DCM. The combined extracts were dried and concentrated, and the residue was purified by preparative HPLC to give N-((6-amino-2-cyclopropylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (7.7 mg, 13%) as a white solid. LRMS (M+H ⁺ )m/z calculated value 444.2, found value 444.1.

^1H NMR(DMSO-d6,400MHz)δ9.00(t,1H),8.82(d,1H),8.62(d,1H),8.51(d,1H),7.97(d,2H),7.85(s,1H),7.72(d,2H ),7.61(d,2H),7.19(d,1H),6.15(d,2H),5.61(s,2H),4.42(d,2H),4.36(s,2H),0.83(s,2H),0.71-0.74(m,2H).

Example 44: Preparation of 2-((3-chloroquinolin-6-yl)methyl)-N-((6-(dimethylamino)-2-methylpyridin-3-yl)methyl)isonicotinamide

2-((3-chloroquinolin-6-yl)methyl)-N-((6-(dimethylamino)-2-methylpyridin-3-yl)methyl)isonicotinamide

Step 1: Preparation of 6-(dimethylamino)-2-methylnicotinonitrile

To a solution of 6-amino- ₂ -methyl-nicotinonitrile (2 g, 15 mmol, 1.0 eq) and _CH3I (21 mg, 150 mmol, 10 eq) in THF (10 mL) was added NaH (1.8 mg, 75 mmol, 5.0 eq) under N2. The mixture was stirred at room temperature overnight. The mixture was quenched with water and extracted with DCM. The combined extracts were dried and concentrated in vacuo to provide the compound (2.2 g, 91%), which was used in the next step without further purification.

Step 2: Preparation of 5-(aminomethyl)-N,N,6-trimethylpyridin-2-amine

To a solution of 6-dimethylamino-2-methyl-nicotinonitrile (200 mg, 1.19 mmol, 1.0 equiv) in MeOH (10 mL) was added Raney nickel (400 mg) under _H . The mixture was stirred at room temperature overnight. The reaction mixture was filtered and the filtrate was concentrated to provide the product (180 mg, 91%), which was used directly in the next step without further purification.

Step 3: Preparation of 2-((3-chloroquinolin-6-yl)methyl)-N-((6-(dimethylamino)-2-methylpyridin-3-yl)methyl)isonicotinamide

To a solution of 2-(3-chloro-quinolin-6-ylmethyl)-isonicotinic acid (326 mg, 1.09 mmol, ₁ eq) in DMF (10 mL) was added (5-aminomethyl-6-methyl-pyridin-2-yl)-dimethylamine (180 mg, 1.09 mmol, 1 eq), HATU (497 mg, 1.3 mmol, 1.2 eq) and Et3N (1 mL). The reaction mixture was stirred at room temperature for 3 h. It was then quenched with water and extracted with DCM. The combined extracts were dried and concentrated. The residue was purified by preparative HPLC to give 2-((3-chloroquinolin-6-yl)methyl)-N-((6-(dimethylamino)-2-methylpyridin-3-yl)methyl)isonicotinamide (130 mg, 30%) as a gray solid. LRMS (M+H ⁺ ) m/z calculated value 446.2, found value 445.8.

¹ H NMR(DMSO,400MHz)δ9.00-9.02(m,1H),8.82(d,1H),8.62(d,1H),8.51(d,1H),7.96-7.98(m,1H),7.84(s, 1H),7.72-7.75(m,2H),7.61-7.62(m,1H),7.34-7.37(m,1H),6.40-6.41(m,1H),4.31(d,4H),2.96(s,6H),

2.35(s,3H).

Example 45: Preparation of 2-((2-(aminomethyl)quinolin-6-yl)methyl)-N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)isonicotinamide

2-((2-(Aminomethyl)quinolin-6-yl)methyl)-N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)isonicotinamide (60 mg, 79%) was prepared as described for Example 146. LRMS (M+H ⁺ ) m/z calcd 474.1, found 474.1. ¹ HNMR(DMSO-d ₆ ,400MHz)δ11.53(s,1H),9.58(s,1H),8.83(d,1H),8.60(br,1H),8.43(d,1H),8.10(d,1H),8.02-8.00(m,3H),7.85(d,1H),7.6 2(d,1H),7.51(s,1H),7.47(d,1H),7.44(d,1H),7.31(s,1H),7.24(d,1H),7.20(d,1H),4.60(d,2H),4.57(s,2H),4.40(q,2H).

Example 46: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((2-(aminomethyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((2-(aminomethyl)quinolin-6-yl)methyl)isonicotinamide (35 mg, 53%) was prepared as described for Example 146. LRMS (M+H ⁺ ) m/z calcd 427.2, found 427.2. ¹ H NMR (CD ₃ OD,400MHz)δ8.99(d,1H),8.69(d,1H),8.39(s,1H),8.35(d,1H),8.25(d,1H),8.16(s,1H),7.98( d,1H),7.80(d,1H),6.74(s,1H),4.82(s,2H),4.63(s,2H),4.59(s,2H),2.63(s,3H),2.50(s,3H).

Example 47: Preparation of N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide

N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of 5-(aminomethyl)benzo[d]isoxazol-3-amine bis(tert-butyl carbamate)

To a mixture of N-hydroxy-acetamide (964 mg, 12.86 mmol, 1.5 equivalents) in DMF (40 mL) was added t-BuOK (1.4 g, 12.86 mmol, 1.5 equivalents). After stirring at room temperature for 30 min, 4- (aminomethyl) -2- fluorobenzonitrile bis (tert-butyl carbamate) (3 g, 8.57 mmol, 1.0 equivalents) was added. The reaction mixture was stirred at room temperature for 5 h and then concentrated. The residue was purified by column chromatography on silica gel (PE/EtOAc=4/1 to 3/1, v/v) to obtain 5- (aminomethyl) benzo [d] isoxazole -3- amine bis (tert-butyl carbamate) (2 g, 64%) as a white solid. LRMS (M+H ⁺ ) m/z calculated value 364 found 364.

Step 2: Preparation of 5-aminomethyl-benzo[d]isoxazol-3-ylamine dihydrochloride

To a mixture of 5-(aminomethyl)benzo[d]isoxazol-3-amine bis(tert-butylcarbamate) (2 g, 5.51 mmol, 1.0 equiv) in MeOH (20 mL) was added 3N HCl in EtOAc (5 mL). After stirring at room temperature for 2 h, the reaction mixture was filtered and the filter cake was washed with _Et2O to give crude 5-aminomethyl-benzo[d]isoxazol-3-ylamine dihydrochloride (1.5 g) as a white solid. LRMS (M+H ⁺ ) m/z calcd 164 found 164.

Step 3: Preparation of N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(3-methyl-quinolin-6-ylmethyl)-isonicotinic acid (150 mg, crude) in DMF (15 mL) was added 5-aminomethyl-benzo[d]isoxazol-3-ylamine dihydrochloride (80 mg, 0.34 mmol, 1.0 equiv), followed by EDCI (98 mg, 0.51 mmol, 1.5 equiv), HOBT (69 mg, 0.51 mmol, 1.5 equiv), and TEA (103 mg, 1.02 mmol, 3.0 equiv). The reaction mixture was heated to 45° C. and stirred overnight. Water was added, and the mixture was extracted with DCM. The organic layer was washed with water, dried over Na ₂ SO ₄ , filtered, and concentrated. The residue was purified by preparative HPLC to give N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (30 mg, 21%) as a yellow solid. LRMS (M+H ⁺ ) m/z calcd 424.2, found 424.0.

¹ H NMR (DMSO-d ₆ ,400MHz): δ9.37(t,1H),8.71(s,1H),8.66(d,1H),8.06(s,1H),7.89(d,1H),7.74-7.78(m,3H),7.6 6(d,1H),7.61(d,1H),7.35(s,1H),7.21(d,1H),6.37(s,2H),4.58(d,2H),4.35(s,2H),2.46(s,3H).

Example 48: Preparation of N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide

N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide

N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide (30 mg, 21%) was prepared as a yellow solid as described for N-((3-aminobenzo[d]isoxazol-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 47). LRMS (M+H ⁺ ) m/z calcd 424.2, found 423.9. ¹ HNMR(DMSO-d ₆ ,400MHz)δ9.40(t,1H),8.81(s,1H),8.66(d,1H),8.10(s,1H),7.89(d,1H),7.82(s,1H),7.76(d,1H),7 .68(t,2H),7.55(d,1H),7.35(s,1H),7.22(d,1H),6.38(s,2H),4.60(d,2H),4.37(s,2H),2.48(s,3H).

Example 49: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of methyl 3-chloro-8-iodoquinoline-6-carboxylate

To a solution of methyl 8-iodoquinoline-6-carboxylate (30 g, 96 mmol, 1.0 eq) in AcOH (1.0 L) was added NCS (38 g, 293 mmol, 3 eq). The mixture was stirred at 100 ° C. overnight, the mixture was concentrated in vacuo, and the residue was purified by silica gel chromatography (PE/DCM=1/1, v/v) to give methyl 3-chloro-8-iodoquinoline-6-carboxylate (15 g, 49%) as a yellow solid.

Step 2: Preparation of (3-chloro-8-iodo-quinolin-6-yl)-methanol

To a solution of 3-chloro-8-iodoquinoline-6-methyl formate (12 g, 34.5 mmol, 1.0 equivalent) in anhydrous THF (200 mL) was carefully added lithium tri-tert-butoxyaluminum hydride (22 g, 70 mmol, 3.4 equivalents). The mixture was stirred at 50 ° C for 5 h under _N protection. EtOAc and water were then added. The organic layer was concentrated and purified by silica gel chromatography (PE/DCM=1/1, v/v) to obtain (3-chloro-8-iodo-quinoline-6-yl)-methanol (7.6 g, 69%) as a white solid.

Step 3: Preparation of 3-chloro-6-hydroxymethyl-quinoline-8-carbonitrile

To a solution of (3-chloro-8-iodo-quinolin-6-yl)-methanol (7.6 g, 23.8 mmol, 1.0 equiv) in DMF (100 mL) was carefully added Zn(CN) ( _2.79 g, 23.8 mmol, 1.0 equiv) and _Pd ( _PPh ) (2.75 g, 2.38 mmol, 0.1 equiv). The mixture was stirred at 50 ° C. overnight under _N protection. EtOAc and water were then added. The organic layer was concentrated and purified by silica gel chromatography (PE/DCM=1/2, v/v) to give 3-chloro-6-hydroxymethyl-quinoline-8-carbonitrile (5.0 g, 96%) as a yellow solid.

Step 4: Preparation of 3-chloro-6-chloromethyl-quinoline-8-carbonitrile

A mixture of 3-chloro-6-hydroxymethyl-quinoline-8-carbonitrile (2.9 g, 13.3 mmol, 1.0 equiv) in _SOCl2 (50 mL) was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DCM and treated with saturated _NaHCO3 solution to give 3-chloro-6-chloromethyl-quinoline-8-carbonitrile (2.2 g, 70%) as a yellow solid.

Step 5: Preparation of methyl 2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinate

To a solution of 3-chloro-6-chloromethyl-quinoline-8-carbonitrile (2.0 g, 8.47 mmol, 1.0 equiv) in dioxane (40 mL) was added methyl 2-(trimethylstannyl)isonicotinate (2.8 g, 9.32 mmol, 1.1 equiv) and Pd(PPh ₃ ) ₂ Cl ₂ (597 mg, 0.85 mmol, 0.1 equiv). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, concentrated, and purified by silica gel chromatography (DCM/MeOH=100/1, v/v) to give methyl 2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinate (1.4 g, 49%) as a yellow solid.

Step 6: Preparation of 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinic acid

To a solution of methyl 2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinate (1.4 g, 4.2 mmol, 1.0 equiv) in THF (5 mL) and H ₂ O (5 mL) was added NaOH (200 mg, 5 mmol, 1.2 equiv). The mixture was stirred at room temperature for 2 h. It was then acidified with 1N HCl to pH = 6 and extracted with EtOAc. The organic layer was concentrated to give 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinic acid (1.1 g, 37%) as a white solid.

Step 7: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(3-chloro-quinolin-6-ylmethyl)-isonicotinic acid (120 mg, 0.37 mmol, 1.0 equiv) and (3-chloro-6-fluoro-1H-indol-5-yl)-methylamine hydrochloride (200 mg, 0.73 mmol, 2.0 equiv) in DMF (10 mL) was added HATU (170 mg, 4.4 mmol, 1.2 equiv) and _Et3N (1.0 mL, 7.1 mmol, 19 equiv). The mixture was stirred at room temperature overnight, then EtOAc and water were added. The organic layer was concentrated, and the residue was purified by preparative HPLC to give N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide (170 mg, 91%) as a yellow solid. LRMS (M+H ⁺ ) m/z calcd. 504.1, found 503.8.

¹ H NMR(DMSO-d6,400MHz)δ11.39(s,1H),9.24(m,1H),9.03-9.04(d,1H),8.71-8.72(d,1H),8.64-8.66(d,1H),8.40(d,1 H),8.19(d,1H),7.84(s,1H),7.66-7.68(d,1H),7.44-7.51(m,2H),7.21-7.24(d,1H),4.59-4.60(d,2H),4.43(s,2H).

Example 50: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide (130 mg, 77%) was prepared as a white solid as described for N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide (Example 49). LRMS (M+H ⁺ ) m/z calcd 456.2, found 456.8.

¹ H NMR (DMSO-d6, 400MHz) δ9.03-9.04 (d, 1H), 8.72-8.73 (d, 1H), 8.59-8.66 (m, 3H), 8.38-8.39 (d, 1H), 8.18-8.19 (d, 1H) ,7.79(s,1H),7.60-7.62(dd,1H),6.15(s,1H),5.77(s,1H),4.41(s,2H),4.34-4.35(d,2H),2.31(s,3H),2.18(s,3H).

Example 51: Preparation of N-((3-chloro-4-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-4-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((3-Chloro-4-fluoro-lH-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide (80 mg, 43%) was prepared as a white solid as described for N-((3-chloro-6-fluoro-lH-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide (Example 49). LRMS (M+H ⁺ ) m/z calcd 504.1, found 503.8.

^1H NMR(DMSO-d6,400MHz)δ11.58(s,1H),9.22(m,1H),9.03-9.04(d,1H),8.71-8.72(d,1H),8.63-8.65(d,1H),8.39-8.40( d,1H),8.19(s,1H),7.83(s,1H),7.65-7.66(m,1H),7.50-7.51(d,2H),7.14-7.21(m,2H).4.57-4.59(d,2H),4.42(s,1H)

Example 52: Preparation of 2-((3-chloro-8-cyanoquinolin-6-yl)methyl)-N-((6-fluoro-1H-indol-5-yl)methyl)isonicotinamide

2-((3-chloro-8-cyanoquinolin-6-yl)methyl)-N-((6-fluoro-1H-indol-5-yl)methyl)isonicotinamide

N-((3-Chloro-4-fluoro-lH-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide (80 mg, 46%) was prepared as a white solid as described for N-((3-chloro-6-fluoro-lH-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide (Example 49). LRMS (M+H ⁺ ) m/z calcd 469.7, found 469.7.

¹ H NMR(DMSO-d6,400MHz)δ11.09(s,1H),9.19(m,1H),9.03-9.04(d,1H),8.72-8.73(d,1H),8.64-8.65(d,1H),8.39-8.40(d,1H),8.19(s,1 H),7.84(s,1H),7.67-7.69(m,1H),7.49-7.50(d,2H),7.31-7.32(t,1H)7.14-7.18(m,2H),6.39(s,1H),4.56-4.58(d,2H),4.42(s,1H).

Example 53: Preparation of N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide (120 mg, 73%) was prepared as a yellow solid as described for N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide (Example 49). LRMS (M+H) m/z calcd 443.1, found 443.0.

¹ H NMR (DMSO-d6, 400MHz) δ9.03-9.04 (d, 1H), 8.92-8.93 (t, 1H), 8.72-8.73 (d, 1H), 8.61-8.63 (m, 3H), 8.39-8.40 (d, 1H), 8. 18-8.19 (d, 1H), 7.79 (s, 2H), 7.61-7.63 (dd, 1H), 6.27 (s, 1H), 5.77 (s, 1H), 4.41 (s, 2H), 4.29-4.32 (d, 2H), 2.15 (s, 3H).

Example 54: Preparation of 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-N-(5-chloro-1H-indazol-3-ylmethyl)-isonicotinamide

2-(3-Chloro-8-cyano-quinolin-6-ylmethyl)-N-(5-chloro-1H-indazol-3-ylmethyl)-isonicotinamide

To a solution of 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinic acid (100 mg, 0.31 mmol, 1.0 equiv) in DCM (8 mL) was added HOBT (53 mg, 0.39 mmol, 1.3 equiv), EDCI (86 mg, 0.45 mmol, 1.5 equiv), _Et3N (0.13 mL, 0.9 mmol, 3.0 equiv) and (5-chloro-1H-indazol-3-yl)-methylamine (67 mg, 0.37 mmol, 1.2 equiv). The mixture was stirred at room temperature for 12 h and diluted with water. The organic layer was separated and the aqueous layer was extracted with DCM. The combined extracts were dried and concentrated. The residue was purified by flash chromatography on a silica gel column (DCM/MeOH=10/1, v/v) to give 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-N-(5-chloro-1H-indazol-3-ylmethyl)-isonicotinamide (50 mg, 34%) as a white solid. LRMS (M+H ⁺ ) m/z calcd 487.1, found 486.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ13.07(s,1H),9.36(m,1H),9.04(d,1H),8.70(d,1H),8.63(d,1H),8.38(d,1H),8.18(s,1 H),7.89(d,1H),7.82(s,1H),7.64(d,1H),7.53(d,1H),7.31-7.34(d,1H),4.78(d,2H),4.42(s,2H).

Example 55: Preparation of 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-N-(3-chloro-1H-pyrrolo[2,3-b]pyridin-5-ylmethyl)-isonicotinamide

2-(3-Chloro-8-cyano-quinolin-6-ylmethyl)-N-(3-chloro-1H-pyrrolo[2,3-b]pyridin-5-ylmethyl)-isonicotinamide

2-(3-Chloro-8-cyano-quinolin-6-ylmethyl)-N-(3-chloro-1H-pyrrolo[2,3-b]pyridin-5-ylmethyl)-isonicotinamide (90 mg, 66%) was prepared as a white solid as described for 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-N-(5-chloro-1H-indazol-3-ylmethyl)-isonicotinamide (Example 54). LRMS (M+H ⁺ ) m/z calcd 487.1, found 486.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.95(s,1H),9.34(m,1H),9.04(d,1H),8.71(d,1H),8.64(d,1H),8.39(d,1H),8.31( s,1H),8.19(s,1H),7.87(s,1H),7.83(d,1H),7.65-7.67(m,2H),4.59-4.61(d,2H),4.42(s,2H).

Example 56: Preparation of N-(6-amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinamide

N-(6-Amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinamide

N-(6-Amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinamide (50 mg, 38%) was prepared as a white solid as described for 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-N-(5-chloro-1H-indazol-3-ylmethyl)-isonicotinamide (Example 54). LRMS (M+H ⁺ ) m/z calcd 441.9, found 441.9.

¹ H NMR (DMSO-d ₆ , 400MHz) δ9.04 (m, 2H), 8.72 (d, 1H), 8.64 (d, 1H), 8.39 (d, 1H), 8.19 (s, 1H), 7.80 (s ,1H),7.63(d,1H),7.38(m,1H),6.38(m,1H),6.25(m,1H),4.42(s,2H),4.30(d,2H).

Example 57: Preparation of N-(1-amino-isoquinolin-6-ylmethyl)-2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinamide

N-(1-Amino-isoquinolin-6-ylmethyl)-2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinamide

N-(1-Amino-isoquinolin-6-ylmethyl)-2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinamide (100 mg, 73%) was prepared as a white solid as described for 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-N-(5-chloro-1H-indazol-3-ylmethyl)-isonicotinamide (Example 54). LRMS (M+H ⁺ ) m/z calcd 479.1, found 479.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.39(m,2H),9.04(d,1H),8.72(d,1H),8.66(d,1H),8.40(s,1H),8.20(s,1H),8.12-8.15(d,1H),7.86(s,1H),7.7 5-7.76 (d, 1H), 7.69-7.70 (d, 1H), 7.56 (s, 1H), 7.39-7.41 (d, 1H), 6.84-6.85 (d, 1H), 6.72 (s, 1H), 4.62 (d, 2H), 4.44 (s, 2H).

Example 58: Preparation of 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide

6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide

To a solution of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-cyanoquinolin-6-yl)methyl)isonicotinamide hydrochloride (100 mg, 0.22 mmol, 1.0 _equiv ) and _K2CO3 (215 mg, 1.56 mmol, 7.3 equiv) in DMSO (10 mL) was added _H2O2 (1 _mL ). The mixture was stirred at 50 °C for 3 h, then EtOAc and water were added. The organic layer was concentrated and purified by preparative HPLC to give 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (71 mg, 48%) as a white solid. LRMS (M+H+) m/z calcd 475.2, found 474.8.

^1H NMR (DMSO-d6, 400MHz) δ9.62 (d, 1H), 8.96-8.98 (d, 1H), 8.59-8.70 (m, 3H), 8.43-8.48 (d, 1H), 8.03-8.04 (d, 1H), 7.94 (s, 1H), 7.79(s, 1H), 7.60-7.62(dd, 1H), 6.11(s, 1H), 5.66(s, 1H), 4.40(s, 2H), 4.33-4.35(d, 2H), 2.30(s, 3H), 2.16(s, 3H).

Example 59: Preparation of 3-chloro-6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide (30 mg, 58%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (Example 58). LRMS (M+H+) m/z calcd 522.1, found 521.9.

^1H NMR(DMSO-d6,400MHz)δ11.39(s,1H),9.63(s,1H),9.24-9.26(t,1H),8.97-8.98(d,1H),8.64-8.70(dd,2H),8.44-8.45(d,1H) ,8.04-8.05(d,1H),7.95-7.96(d,1H),7.66-7.68(d,1H),7.43-7.50(m,1H),7.20-7.24(d,1H),4.57-4.59(d,2H),4.42(s,2H).

Example 60: Preparation of 3-chloro-6-((4-(((5-chloro-1H-indazol-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-((5-chloro-1H-indazol-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-(((5-chloro-1H-indazol-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide (20 mg, 66%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (Example 58). LRMS (M+H) m/z calcd 505.1, found 504.9.

¹ H NMR (DMSO-d ₆ ,400MHz)δ13.09(s,1H),9.63(s,1H),9.40-9.41(t,1H),8.97-8.98(d,1H),8.64-8.70(dd,2H),8.43(s,1H) ,7.84-8.02(m,3H),7.64-7.65(d,1H),7.51-7.54(d,1H),7.31-7.32(m,1H),4.77-4.79(d,2H),4.41(s,2H).

Example 61: Preparation of 3-chloro-6-((4-(((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-(((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-(((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide (30 mg, 37%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (Example 58). LRMS (M+H+) m/z calcd 505.1, found 505.0.

¹ H NMR (DMSO-d ₆ , 400MHz) δ9.63 (s, 1H), 9.34-9.35 (t, 1H), 8.96-8.97 (d, 1H), 8.69-8.70 (d, 1H), 8.64-8.66 (d, 1H), 8.44 (s, 1H), 8 .31(s,1H),8.04(s,1H),7.95(s,1H),7.87(s,1H),7.82(s,1H),7.65-7.67(d,2H),4.58-4.60(d,2H),4.42(s,2H).

Example 62: Preparation of 6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide

6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide

6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (28 mg, 44%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (Example 58). LRMS (M+H) m/z calcd 461.1, found 461.1.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.63(s,1H),8.97-9.00(m,2H),8.70-8.71(d,1H),8.62-8.64(d,1H),8.44-8.46(d,1H),8.04(s,1H),7.96(s,1 H),7.63.-7.65(dd,1H),7.23-7.25(d,1H),6.22-6.25(dd,1H),5.77(s,1H),4.42(s,2H),4.28-4.30(d,2H),2.28(s,3H).

Example 63: Preparation of 6-((4-(((6-amino-4-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide

6-((4-((6-amino-4-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide

6-((4-(((6-amino-4-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (70 mg, 69%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (Example 58). LRMS (M+H) m/z calcd 461.1, found 461.1.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.63(s,1H),8.92-9.00(m,2H),8.70-8.71(d,1H),8.62-8.64(d,1H),8.43-8.45(d,1H),8.05(s,1H),7.96( s,1H),7.78.-7.80(dd,1H),7.61-7.63(d,1H),6.26(s,1H),5.81(s,1H),4.41(s,2H),4.29-4.31(d,2H),2.15(s,3H).

Example 64: Preparation of 6-((4-(((1-aminoisoquinolin-6-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide

6-((4-(((1-aminoisoquinolin-6-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide

6-((4-(((1-aminoisoquinolin-6-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (25 mg, 16%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (Example 58). LRMS (M+H) m/z calcd 497.1, found 497.0.

¹ H NMR (DMSO-d ₆ , 400MHz) δ9.63 (s, 1H), 9.37-9.39 (t, 1H), 8.97-8.98 (m, 2H), 8.70-8.71 (d, 1H), 8. 67-8.68(d,1H),8.45-8.46(d,1H),8.13-8.15(d,1H),8.06(s,1H),7.95(s,1H),7. 85(s, 1H), 7.75-7.76(d, 1H), 7.69-7.70(dd, 1H), 7.54-7.56(dd, 1H), 7.42(s, 1H), 7.39-7.40 (d, 1H), 6.84-6.86 (d, 1H), 6.76 (s, 2H), 4.60-4.62 (d, 2H), 4.40 (s, 2H).

Example 65: Preparation of 3-chloro-6-((4-(((3-chloro-4-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-(((3-chloro-4-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-(((3-chloro-4-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide (35 mg, 16%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (Example 58). LRMS (M+H) m/z calcd 522.1, found 522.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.58(s,1H),9.62(s,1H),9.22-9.25(t,1H),8.97-8.98(d,2H),8.69-8.70(d,1H),8.63-8.65(d,1H),8.43-8.44(d ,1H),8.04(s,1H),7.94(s,1H),7.82(s,1H),7.65-7.66(d,1H),7.51(s,1H),7.14-7.20(m,2H),4.56-4.57(d,2H),4.41(s,2H).

Example 66: Preparation of 3-chloro-6-((4-(((6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-((6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide

3-Chloro-6-((4-(((6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)quinoline-8-carboxamide (35 mg, 42%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide (Example 58). LRMS (M+H) m/z calcd 488.1, found 488.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.09(s,1H),9.62(s,1H),9.19-9.22(t,1H),8.97-8.98(d ,2H),8.69-8.70(d,1H),8.64-8.66(d,1H),8.44-8.45(d,1H),8.05(s ,1H),7.95(s,1H),7.84(s,1H),7.67-7.69(d,1H),7.48-7.50(s,1H), 7.30-7.31(t,2H),7.14-7.17(1,1H),4.55-4.57(d,2H),4.42(s,2H).

Example 67: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of (3-chloro-8-methanesulfonyl-quinolin-6-yl)-methanol

A mixture of (3-chloro-8-iodo-quinolin-6-yl)-methanol (7.6 g, 23.8 mmol, 1 eq), sodium methanesulfinate (2.92 g, 28.6 mmol, 1.2 eq), copper iodide (452 mg, 2.38 mol, 0.1 eq), and L-proline sodium salt (652 mg, 4.76 mol, 0.2 eq) in 110 mL of DMSO was heated to 110° C. under nitrogen for 15 h. The cooled mixture was partitioned between ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with brine, dried over MgSO ₄ , and concentrated in vacuo. The residue was purified by silica gel column purification (EtOAc/PE=1/2, v/v) to give (3-chloro-8-methanesulfonyl-quinolin-6-yl)-methanol (4.1 g, 64%) as a yellow solid.

Step 2: Preparation of 3-chloro-6-chloromethyl-8-methylsulfonyl-quinoline

To (3-chloro-8-methanesulfonyl-quinolin-6-yl)-methanol (4.1 g, 15.1 mmol, 1.0 equiv) was added _SOCl₂ (50 mL) and the mixture was stirred at room temperature for 1 h. The volatiles were removed under vacuum and the residue was dissolved in DCM. The mixture was washed with saturated aqueous _NaHCO₃ , dried and concentrated to give 3-chloro-6-chloromethyl-8-methanesulfonyl-quinoline (4.3 g, 99%) as a yellow solid.

Step 3: Preparation of methyl 2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinate

To a solution of 3-chloro-6-chloromethyl-8-methanesulfonyl-quinoline (4.3 g, 14.9 mmol, 1.0 equiv) in dioxane (70 mL) was added methyl 2-(trimethylstannyl)isonicotinate (4.93 g, 16.4 mmol, 1.1 equiv) and Pd( _PPh3 ) _2Cl2 (1.04 g, 1.49 mmol, 0.1 equiv). The mixture was stirred at 90°C for 3 h under a nitrogen atmosphere, concentrated, and purified by silica _gel chromatography (DCM/MeOH=50/1, v/v) to give methyl 2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinate (2.3 g, 40%) as a yellow solid.

Step 4: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(3-chloro-8-methanesulfonyl-quinolin-6-ylmethyl)-isonicotinic acid (80 mg, 0.21 mmol, 1 eq) in DMF (10 mL) was added 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine hydrochloride (40 mg, 0.21 mmol, 1.0 eq), followed by EDCI (61 mg, 0.32 mmol, 1.5 eq), HOBT (43 mg, 0.32 mmol, 1.5 eq), and TEA (64 mg, 0.64 mmol, 3.0 eq). The reaction mixture was heated to 45° C. and stirred overnight. Water was added, and the mixture was extracted with DCM. The organic layer was washed with water, dried over Na ₂ SO ₄ , filtered, and concentrated. The residue was purified by preparative HPLC to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (40 mg, 37%) as a yellow solid. LRMS (M+H ⁺ ) m/z calcd 510.1, found 509.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.07(d,1H),8.76(d,1H),8.66(t,1H),8.62(d,1H),8.36(d,1H),8.23(s,1H),7.83(s,1H),7 .62(d,1H),6.12(s,1H),5.70(s,2H),4.47(s,2H),4.34(d,2H),3.56(s,3H),2.30(s,3H),2.17(s,3H).

Example 68: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((3-Chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (40 mg, 34%) was prepared as a white solid as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (Example 67). LRMS (M+H ⁺ ) m/z calcd 557.1, found 557.0.

¹ H NMR (DMSO-d ₆ ,400MHz) δ11.41(s,1H),9.27(t,1H),9.07(d,1H),8.76(d,1H),8.67(d,1H),8.37(s,1H),8.23(s,1H), 7.88(s, 1H), 7.68(d, 1H), 7.51(d, 2H), 7.46(d, 1H), 7.23(d, 2H), 4.59(d, 2H), 4.49(s, 2H), 3.56(s, 3H).

Example 69: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (Example 67) was prepared as a white solid (40 mg, 34%). LRMS (M+H ⁺ ) m/z calcd 496.1, found 495.7.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.07(d,1H),9.01(t,1H),8.76(d,1H),8.64(d,1H),8.36(d,1H),8.23(s,1H),7.85(s,1H),7 .65(d,1H),7.24(d,1H),6.23(d,1H),5.75(s,2H),4.48(s,2H),4.30(d,2H),3.56(s,3H),2.28(s,3H).

Example 70: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

To a solution of methyl 2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinate (300 mg, 0.95 mmol, 1.0 equiv) in THF (16 mL)/H ₂ O (4 mL) was added LiOH.H ₂ O (79.49 mg, 1.89 mmol, 2.0 equiv). The mixture was stirred at 40° C. for 1 h and acidified to pH 5 with 1N HCl solution. The mixture was concentrated in vacuo and the residue was used directly without further purification. To a solution of the above crude product and 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine (285 mg, 1.89 mmol, 2.0 equiv) in DMF (10 mL) was added HOBT (192.37 mg, 1.43 mmol, 1.5 equiv), EDCI (310.08 mg, 1.62 mmol, 1.7 equiv) and _Et3N (0.53 mL, 3.8 mmol, 4 equiv). The mixture was stirred at room temperature overnight and diluted with water. The organic layer was separated and the aqueous layer was extracted with DCM. The combined extracts were dried and concentrated. The residue was purified by preparative HPLC to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (90 mg, 21% over 2 steps) as an off-white solid. LRMS (M+H ⁺ ) m/z calcd. 437.2, found 437.0.

¹ H NMR (DMSO-d ₆ , 400MHz) δ8.89 (s, 1H), 8.61 (t, 2H), 8.23 (s, 2H), 8.10 (s, 1H), 7.78 (s, 1H), 7.61 (t, 1H), 6 .12(s,1H),5.67(s,2H),4.37(s,2H),4.35(d,2H),2.50(s,3H),2.30(s,3H),2.16(s,3H).

Example 71: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (105 mg, 24% over 2 steps) was prepared as an off-white solid as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (Example 70). LRMS (M+H ⁺ ) m/z calcd 459.2, found 459.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.41(t,1H),8.90(s,1H),8.69(d,1H),8.28(d,2H),8.16(d,2H),7.86(s,1H),7.78(d,1H),7 .71 (d, 1H), 7.57 (s, 1H), 7.43 (d, 1H), 6.87 (d, 1H), 6.78 (s, 2H), 4.64 (d, 2H), 4.42 (s, 2H), 2.50 (s, 3H).

Example 72: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((3-Chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (100 mg, 21% over 2 steps) was prepared as an off-white solid as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (Example 70). LRMS (M+H ⁺ ) m/z calcd 484.1, found 483.9.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.32(s,1H),8.90(t,1H),8.67(d,1H),8.62(d,1H),8.27(d,2H),8.12(s,1H),7.83(s ,1H),7.68(d,1H),7.52(d,1H),7.46(d,1H),7.25(d,1H),4.61(d,2H),4.41(s,2H),2.50(s,3H).

Example 73: Preparation of N-((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(8-cyano-3-methyl-quinolin-6-ylmethyl)-isonicotinic acid (100 mg, 0.33 mmol, 1.0 eq) and (3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)-methylamine hydrochloride (144 mg, 0.66 mmol, 2.0 eq) in DMF (5 mL) was added HATU (188 mg, 0.50 mmol, 1.5 eq) and _Et3N (134 mg, 1.32 mmol, 4 eq). The mixture was stirred at room temperature for 2 h and diluted with water. The organic layer was separated and the aqueous layer was extracted with DCM. The combined extracts were dried and concentrated. The residue was purified by preparative HPLC to give N-((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (66 mg, 43%) as an off-white solid. LRMS (M+H ⁺ ) m/z calcd 467.1, found 466.8. ¹ H NMR (DMSO-d ₆ , 400MHz) δ: 11.97 (s, 1H), 9.34 (t, 1H), 8.89 (s, 1H), 8.66 (d, 1H), 8.32 (s, 1H), 8.26 (s, 1H), 8.23 (s, 1H), 8.11 (s, 1H), 7.88 (s, 1H), 7.82 (s, 1H), 7.68 (d, 1H), 7.66 (d, 1H), 4.61 (d, 2H), 4.40 (s, 2H), 2.50 (s, 3H).

Example 74: Preparation of N-((3-chloro-4-fluoro-1H-indol-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-4-fluoro-1H-indol-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((3-Chloro-4-fluoro-lH-indol-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (70 mg, 44%) was prepared as an off-white solid as described for N-((3-chloro-lH-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (Example 73). LRMS (M+H ⁺ ) m/z calcd 484.1, found 483.8.

¹ H NMR (DMSO-d ₆ , 400MHz) δ11.59 (s, 1H), 9.23 (t, 1H), 8.80 (d, 1H), 8.65 (d, 1H), 8.25 (s, 1H), 8.12 (s, 1H), 7.81 (s , 1H), 7.65(d, 1H), 7.52(d, 1H), 7.20(d, 1H), 7.15(d, 1H), 4.58(d, 2H), 4.39(s, 2H), 2.50(s, 3H).

Example 75: Preparation of N-((5-chloro-1H-indazol-3-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((5-chloro-1H-indazol-3-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide

N-((5-Chloro-1H-indazol-3-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (74 mg, 48%) was prepared as an off-white solid as described for N-((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (Example 73). LRMS (M+H ⁺ ) m/z calcd 467.1, found 466.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ13.09(s,1H),9.38(t,1H),8.90(d,1H),8.65(d,1H),8.25(s,2H),8.12(s,1H),7.89(s,1H), 7.81(s,1H),7.64(d,1H),7.53(d,1H),7.34(d,1H),7.32(d,1H),4.79(d,2H),4.39(s,2H),2.50(s,3H).

Example 76: Preparation of 2-((8-cyano-3-methylquinolin-6-yl)methyl)-N-((6-fluoro-1H-indol-5-yl)methyl)isonicotinamide

2-((8-cyano-3-methylquinolin-6-yl)methyl)-N-((6-fluoro-1H-indol-5-yl)methyl)isonicotinamide

2-((8-Cyano-3-methylquinolin-6-yl)methyl)-N-((6-fluoro-1H-indol-5-yl)methyl)isonicotinamide (70 mg, 46%) was prepared as an off-white solid as described for N-((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinamide (Example 73). LRMS (M+H ⁺ ) m/z calcd 450.2, found 449.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.11(s,1H),9.20(t,1H),8.90(d,1H),8.65(d,1H),8.26(t,2H),8.13(s,1H),7.83(s ,1H),7.68(d,1H),7.31(d,1H),7.17(d,1H),6.40(s,1H),4.57(d,2H),4.40(s,2H),2.50(s,3H).

Example 77: Preparation of 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

To a solution of N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(8-cyano-3-methyl-quinolin-6-ylmethyl)-isonicotinamide (80 mg, 0.18 mmol, 1.0 equiv) and _{K2CO3 (} 180.7 mg, 1.31 mmol, 7.3 equiv) in DMSO (10 mL) was added _H2O2 (1 _mL ). The mixture was stirred at 50 °C for 3 h, then EtOAc and water were added, the organic layer was concentrated and purified by preparative HPLC to give 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (4 mg, 4.8%) as a white solid. LRMS (M+H+) m/z calcd 455.2, found 454.9.

¹ H NMR (DMSO-d ₆ , 400MHz) δ10.20 (s, 1H), 8.84 (s, 1H), 8.66 (s, 1H), 8.63 (s, 1H), 8.41 (s, 1H), 8.25 (s, 1H), 7.98 (s, 1H), 7.90 (s, 1H), 7.78(s,1H),7.61(s,1H),6.12(s,1H),5.68(s,2H),4.38(d,2H),4.34(s,2H),2.50(s,3H),2.29(s,3H),2.21(s,3H).

Example 78: Preparation of 6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((3-chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((3-Chloro-6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (40 mg, 32%) was prepared as an off-white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (Example 77). LRMS (M+H ⁺ ) m/z calcd 502.1, found 501.8. ¹ H NMR (DMSO-d ₆ ,400MHz)δ11.44(s,1H),10.28(d,1H),9.31(t,1H),8.89(s,1H),8.71(d,1H),8.48(s,1H),8.28(s,1H),8.02(s,1H), 7.93(s,1H),7.86(s,1H),7.72(d,1H),7.55(d,1H),7.50(d,1H),7.29(d,1H),4.64(d,2H),4.45(s,2H),2.54(s,3H).

Example 79: Preparation of 6-((4-(((1-aminoisoquinolin-6-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((1-aminoisoquinolin-6-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((1-aminoisoquinolin-6-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (35 mg, 48%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (Example 77). LRMS (M+H ⁺ ) m/z calcd 477.2, found 476.9.

¹ H NMR (DMSO-d ₆ ,400MHz)δ10.22(s,1H),9.40(t,1H),8.85(d,1H),8.68(d,1H),8.43(s,1H),8.26(s,1H),8.15(d,1H),8.00(s,1H),7.89(d ,1H),7.76(d,1H),7.70(d,1H),7.56(s,1H),7.42(d,2H),6.86(d,1H),6.76(s,2H),4.62(d,2H),4.42(s,2H),2.50(s,3H).

Example 80: Preparation of 6-((4-(((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((3-Chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (20 mg, 38%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (Example 77). LRMS (M+H ⁺ ) m/z calcd 485.1, found 484.8.

¹ H NMR (DMSO-d ₆ , 400MHz) δ10.23 (s, 1H), 9.36 (t, 1H), 8.84 (d, 1H), 8.66 (d, 1H), 8.43 (d, 1H), 8.31 (d, 1H), 8.24 (s, 1H), 7.98 (d, 1H), 7.88 (s, 2H), 7.80 (s, 1H), 7.67 (s, 1H), 7.66 (d, 1H), 4.60 (d, 2H), 4.40 (s, 2H), 2.51 (s, 3H).

Example 81: Preparation of 6-((4-(((3-chloro-4-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((3-chloro-4-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((3-Chloro-4-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (30 mg, 41%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (Example 77). LRMS (M+H) m/z calcd 502.1, found 501.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.59(s,1H),10.23(s,1H),9.26(t,1H),8.85(d,1H),8.65(d,1H),8.42(d,1H),8.25(s,1H),7.98(d,1H), 7.88(d,1H),7.80(s,1H),7.66(d,1H),7.51(d,1H),7.21(d,1H),7.16(d,1H),4.58(d,2H),4.40(s,2H),2.51(s,3H).

Example 82: Preparation of 6-((4-(((5-chloro-1H-indazol-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-((5-chloro-1H-indazol-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((5-Chloro-1H-indazol-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (20 mg, 48%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (Example 77). LRMS (M+H) m/z calcd 485.1, found 484.8.

¹ H NMR (DMSO-d ₆ ,400MHz)δ13.08(s,1H),10.21(s,1H),9.39(s,1H),8.84(s,1H),8.65(d,1H),8.42(s,1H),8.25(s,1H),7.97( s,1H),7.89(s,2H),7.80(s,1H),7.64(d,1H),7.53(d,1H),7.34(d,1H),4.78(d,2H),4.39(s,2H),2.50(s,3H).

Example 83: Preparation of 6-((4-(((6-fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((6-Fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide

6-((4-(((6-Fluoro-1H-indol-5-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (20 mg, 35%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxamide (Example 77). LRMS (M+H) m/z calcd 468.2, found 467.8.

¹ H NMR (DMSO-d ₆ , 400MHz) δ11.11 (s, 1H), 10.23 (s, 1H), 9.23 (t, 1H), 8.84 (s, 1H), 8.66 (d, 1H), 8.44 (s, 1H), 8.26 (s, 1H), 7.99 (s, 1H), 7.90 ( s, 1H), 7.83 (s, 1H), 7.69 (d, 1H), 7.51 (d, 1H), 7.18 (s, 1H), 7.18 (d, 1H), 6.40 (s, 1H), 4.57 (d, 2H), 4.41 (s, 2H), 2.51 (s, 3H).

Example 84: Preparation of 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxylic acid

6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxylic acid

Step 1: Preparation of methyl 6-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxylate

A mixture of methyl 2-((8-cyano-3-methylquinolin-6-yl)methyl)isonicotinate (500 mg, 1.58 mmol, 1 equivalent) in HCl/MeOH (10 M, 25 mL) was heated under reflux for one week. After cooling to room temperature, the solvent was removed by evaporation. The residue was diluted with DCM and washed with saturated NaHCO _3. The organic phase was separated, dried and concentrated. The residue was purified by silica gel column chromatography (EtOAc/PE=2/1, v/v) to give methyl 6-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxylate (100 mg, 18%) as a yellow solid.

Step 2: Preparation of 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxylic acid

To a solution of methyl 6-((4-(methoxycarbonyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxylate (100 mg, 0.31 mmol, 1.0 equiv) in THF/H ₂ O (5 mL, 1:1) was added LiOH.H ₂ O (39 mg, 0.93 mmol, 3 equiv). The mixture was stirred at room temperature for 5 h and then concentrated. To a solution of the above crude product and 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine hydrochloride (88 mg, 0.47 mmol, 1.5 equiv) in DMF (5 mL) was added HATU (188 mg, 0.50 mmol, 1.5 equiv) and Et ₃ N (134 mg, 1.32 mmol, 4 equiv). The mixture was stirred at room temperature for 2 h and diluted with water. The organic layer was separated and the aqueous layer was extracted with DCM, and the combined extracts were dried and concentrated. The residue was purified by preparative HPLC to give 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-methylquinoline-8-carboxylic acid (2 mg, 1.4% over 2 steps) as an off-white solid. LRMS (M+H ⁺ ) m/z calcd 456.2, found 455.9.

¹ H NMR (CD ₃ OD,400MHz)δ8.80(s,1H),8.60(d,1H),8.33(s,1H),8.27(s,1H),7.99(s,1H),7.73(s,1H), 7.31(d,1H),6.32(s,1H),4.49(s,2H),4.43(s,2H),2.57(s,3H),2.39(s,3H),2.26(s,3H).

Example 85: Preparation of N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide

N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide

Step 1: Preparation of dimethoxy-acetic acid

To a solution of ethyl 2,2-dimethoxyacetate (30 g, 170 mmol, 1.0 eq) in THF (100 mL) and H ₂ O (100 mL) was added NaOH (8.2 g, 205 mmol, 1.2 eq). The mixture was stirred at room temperature for 2 h. It was then acidified to pH 6 with 1N HCl and extracted with EtOAc. The organic layer was concentrated to give dimethoxy-acetic acid (15 g, 59%) as a yellow oil.

Step 2: Preparation of N-(3-chloro-benzyl)-2,2-dimethoxy-acetamide

To a solution of dimethoxy-acetic acid (15 g, 100 mmol, 1.1 equiv) and 3-chloro-benzylamine (13 g, 92 mmol, 1.0 equiv) in DMF (200 mL) was added HATU (40 g, 100 mmol, 1.1 equiv) and _Et3N (38 mL, 300 mmol, 3 equiv). The mixture was stirred at room temperature overnight. EtOAc and water were then added, the organic layer was concentrated, and the residue was purified by chromatography on a silica gel column (PE/EtOAc=10/1-1/1, v/v) to give N-(3-chloro-benzyl)-2,2-dimethoxy-acetamide (13.5 g, 50%) as a yellow oil.

Step 3: Preparation of 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinic acid

To a solution of methyl 2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinate (350 mg, 1.1 mmol, 1.0 equiv) in THF (5 mL) and H ₂ O (5 mL) was added NaOH (51 mg, 1.3 mmol, 1.2 equiv). The mixture was stirred at room temperature for 2 h. It was then acidified to pH = 6 with 1N HCl and extracted with EtOAc. The organic layer was concentrated to give 2-(3-chloro-8-cyano-quinolin-6-ylmethyl)-isonicotinic acid (320 mg, 91%) as a white solid.

Step 4: Preparation of N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide

To a solution of 2-(7-chloro-3-oxo-3H-isoquinolin-2-ylmethyl)-isonicotinic acid (50 mg, 0.16 mmol, 1.0 equiv) and 5-aminomethyl-4-methyl-pyridin-2-ylamine hydrochloride (57 mg, 0.33 mmol, 2.0 equiv) in DMF (10 mL) was added HATU (73 mg, 0.19 mmol, 1.2 equiv) and _Et3N (1.0 mL, 7.1 mmol, 44 equiv). The mixture was stirred at room temperature overnight before addition of EtOAc and water. The organic layer was concentrated and the residue was purified by preparative HPLC to give N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide (25 mg, 36%) as a white solid. LRMS (M+H ⁺ ) m/z calcd. 434.1, found 434.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.05(s,1H),9.88-9.90(t,1H),8.68-8.69(d,1H),8.88-8.93(m,1H),7.77(s,1H),7.67 -7.72(m,3H),7.41(d,1H),6.25(s,1H),5.76(s,2H),5.57(s,2H),4.29-4.30(d,2H),2.13(s,3H).

Example 86: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide (Example 85) was prepared as a white solid (27 mg, 37%). LRMS (M+H ⁺ ) m/z calcd 434.1, found 434.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.05-9.07(t,1H),9.04(s,1H),8.69-8.70(t,1H),8.17(d,1H),7.90-7.92(m,1H),7.67-7.63(m,2H ),7.41(s,3H),7.23-7.26(d,1H),6.22-6.24(d,1H),5.76(s,1H),5.57(s,2H),4.28-4.30(d,2H),2.27(s,3H).

Example 87: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide (Example 85) was prepared as a white solid (30 mg, 42%). LRMS (M+H ⁺ ) m/z calcd 448.1, found 448.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ9.04(s,1H),8.71-8.73(t,1H),8.66-8.68(d,1H),8.17-8.18(d,1H),7.87-7.92(m,2H),7.67-7. 70(m,2H),7.40(s,1H),6.11(s,1H),5.70(s,2H),5.56(s,2H),4.33-4.34(d,2H),2.29(s,3H),2.15(s,3H).

Example 88: Preparation of 2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)-N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)isonicotinamide

2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)-N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)isonicotinamide

2-((7-Chloro-3-oxoisoquinolin-2(3H)-yl)methyl)-N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)isonicotinamide (55 mg, 70%) was prepared as a white solid as described for N-((6-amino-4-methylpyridin-3-yl)methyl)-2-((7-chloro-3-oxoisoquinolin-2(3H)-yl)methyl)isonicotinamide (Example 85). LRMS (M+H ⁺ ) m/z calcd 495.1, found 495.0.

¹ H NMR (DMSO-d ₆ ,400MHz)δ11.40(s,1H),9.33-9.35(t,1H),9.04(s,1H),8.72-8.73(d,1H),8.17(s,1H),8.89-8.94(m,2H),7.69-7.7 0 (d, 1H), 7.66-7.67 (d, 1H), 7.50-7.51 (d, 1H), 7.24-7.45 (m, 2H), 7.21-7.24 (d, 1H), 5.59 (D, 2H), 4.58-4.60 (d, 2H).

Example 89: Preparation of N-(6-amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

N-(6-Amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-quinolin-6-ylmethyl)-isonicotinamide

Step 1: Preparation of methyl 8-fluoroquinoline-6-carboxylate

A mixture of methyl 4-amino-3-fluorobenzoate (35 g, 0.207 mmol, 1 eq), acrolein (17.4 g, 0.311 mol, 1.5 eq) and 6N HCl (600 mL) was stirred at 100 ° C for 10 min. The mixture was then cooled and adjusted to pH ~ 5-6 with NaHCO ₃ (aq). The mixture was extracted with DCM. The combined organic layers were washed with brine, dried over MgSO ₄ , filtered, then concentrated and purified by column chromatography (EtOAc/PE=1/20, v/v) to give methyl 8-fluoroquinoline-6-carboxylate (11 g, 21%) as a yellow solid.

Step 2: Preparation of methyl 3-chloro-8-fluoroquinoline-6-carboxylate

To a DMF solution of 8-fluoroquinoline-6-methyl formate (11 g, 53.7 mmol, 1 equivalent) was added NCS (21.4 g, 0.161 mol, 3 equivalents). The reaction mixture was stirred at 120 ° C overnight. The reaction mixture was cooled to ambient temperature, treated with water, neutralized with solid NaHCO ₃ , and further stirred at room temperature for 30 min. Finally, powdered sodium thiosulfate was carefully added to remove excess NCS. The mixture was extracted with EtOAc. The organic layer was dried and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel to obtain 3-chloro-8-fluoroquinoline-6-methyl formate (11.5 g, 90%) as a yellow solid.

Step 3: Preparation of (3-chloro-8-fluoro-quinolin-6-yl)-methanol

To a solution of 3-chloro-8-fluoroquinoline-6-methyl carboxylate (4.5 g, 18.8 mmol, 1 equivalent) was added LiAlH(t-BuO) ₃ (12.0 g, 47.1 mmol, 2.5 equivalents). The resulting mixture was stirred at 40 ° C for 12 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=2/1, v/v) to give (3-chloro-8-fluoro-quinoline-6-yl)-methanol (2.1 g, 53%) as a yellow solid.

Step 4: Preparation of 3-chloro-6-chloromethyl-8-fluoro-quinoline

A mixture of 3-chloro-8-fluoro-6-hydroxymethyl-quinoline (2.1 g, 9.95 mmol, 1.0 equiv) in _SOCl2 (50 mL) was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DCM and treated with saturated _NaHCO3 solution to give 3-chloro-6-chloromethyl-8-fluoro-quinoline (2.2 g, 96%) as a yellow solid.

Step 5: Preparation of methyl 2-((3-chloro-8-fluoroquinolin-6-yl)methyl)isonicotinate

To a solution of 3-chloro-6-chloromethyl-8-fluoro-quinoline (2.2 g, 9.61 mmol, 1.0 equiv) in dioxane (60 mL) was added methyl 2-(trimethylstannyl)isonicotinate (3.18 g, 10.6 mmol, 1.1 equiv) and Pd(PPh ₃ ) ₂ Cl ₂ (674 mg, 0.96 mmol, 0.1 equiv). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, concentrated, and purified by silica gel chromatography (DCM/MeOH=200/1, v/v) to give methyl 2-((3-chloro-8-fluoroquinolin-6-yl)methyl)isonicotinate (1.6 g, 50%) as a yellow solid.

Step 6: Preparation of 2-((3-chloro-8-fluoroquinolin-6-yl)methyl)isonicotinic acid

To a solution of methyl 2-((3-chloro-8-fluoroquinolin-6-yl)methyl)isonicotinate (800 mg, 2.4 mmol, 1 eq) in THF (20 ml)/water (10 ml) was added NaOH (116 mg, 0.29 mmol, 1.2 eq). The mixture was stirred at room temperature for 3 h. Aqueous HCl (2 N) was then added to the reaction mixture until pH 6-7. The mixture was extracted with EtOAc, and the organic layer was concentrated under pressure. The gray compound was used directly in the next step (500 mg, 76%).

Step 7: Preparation of N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(3-chloro-8-fluoro-quinolin-6-ylmethyl)-isonicotinamide

To a solution of 2-(3-chloro-quinolin-6-ylmethyl)-isonicotinic acid (100 mg, 0.3 mmol, 1 equiv) in DMF (10 mL) was added 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine (71 mg, 0.47 mmol, 1.5 equiv), HATU (137 mg, 0.36 mmol, 1.2 equiv) and _Et3N (1 mL). The mixture was stirred at room temperature for 3 h. It was then quenched with water and extracted with DCM. The combined extracts were dried, concentrated, and the residue was purified by preparative HPLC to give N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(3-chloro-8-fluoro-quinolin-6-ylmethyl)-isonicotinamide (39 mg, 29%) as a white solid. LRMS (M+H ⁺ ) m/z calculated value 450.1, found value 449.8.

¹ H NMR(DMSO-d6,400MHz)δ:8.88(d,1H),8.60-8.62(m,3H),7.75(s,1H),7.68(s,1H), 7.58-7.61(m,2H),6.13(s,1H),5.71(s,2H),4.33(s,4H),2.30(s,3H),2.16(s,3H).

Example 90: Preparation of N-(3-chloro-6-fluoro-1H-indol-5-ylmethyl)-2-(3-chloro-8-fluoro-quinolin-6-ylmethyl)-isonicotinamide

N-(3-Chloro-6-fluoro-1H-indol-5-ylmethyl)-2-(3-chloro-8-fluoro-quinolin-6-ylmethyl)-isonicotinamide

N-(3-Chloro-6-fluoro-1H-indol-5-ylmethyl)-2-(3-chloro-8-fluoro-quinolin-6-ylmethyl)-isonicotinamide (47 mg, 20%) was prepared as a white solid as described for N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(3-chloro-8-fluoro-quinolin-6-ylmethyl)-isonicotinamide (Example 89); LRMS (M+H ⁺ ) m/z calcd 497.1, found 497.7.

^1H NMR(DMSO-d6,400MHz)δ11.38(s,1H),9.23(s,1H),8.88(s,1H),8.61-8.66(m,2H),7.80(s,1 H),7.59-7.69(m,3H),7.43-7.50(m,2H),7.20-7.23(d,1H),4.57-4.59(d,2H),4.36(s,2H).

Example 91: Preparation of N-(6-amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-8-fluoro-quinolin-6-ylmethyl)-isonicotinamide

N-(6-Amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-8-fluoro-quinolin-6-ylmethyl)-isonicotinamide

N-(6-Amino-2-methyl-pyridin-3-ylmethyl)-2-(3-chloro-8-fluoro-quinolin-6-ylmethyl)-isonicotinamide (Example 89) was prepared as a white solid (45 mg, 22%). LRMS (M+H ⁺ ) m/z calcd 436.1, found 436.0.

^1H NMR (DMSO-d6, 400MHz) δ8.97 (s, 1H), 8.88 (s, 1H), 8.6-8.64 (m, 2H), 7.77 (s, 1H), 7.68 (s, 1H), 7.59-7.63(m, 2H), 7.42(d, 1H), 6.26(d, 1H), 5.80(d, 2H), 4.35(s, 2H), 4.29(d, 2H).

Example 92: Preparation of methyl 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylate

Methyl 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylate

To a solution of 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine (95 mg, 0.43 mmol, 1.7 eq) in DMF (10 mL) was added 2-((3-chloro-8-(methoxycarbonyl)quinolin-6-yl)methyl)isonicotinic acid (90 mg, 0.25 mmol, 1 eq), HATU (123 mg, 0.32 mmol, 1.3 eq) and _Et3N (0.5 mL). The mixture was stirred at room temperature overnight. It was then quenched with water and extracted with DCM. The combined extracts were dried and concentrated, and the residue was purified by column chromatography (DCM/MeOH=20/1, v/v) to give methyl 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylate (100 mg, 81.9%) as a yellow solid. LRMS (M+H ⁺ ) m/z calcd 490.2, found 490.1.

^1H NMR(DMSO-d6,400MHz)δ8.89-8.90(m,1H),8.59-8.63(m,3H),8.00(s,1H),7.91-7.92(m,1H),7.77(s, 1H),7.60(d,1H),6.15(s,1H),4.37(s,2H),4.33-4.34(d,2H),3.88(s,3H),2.31(s,3H),2.18(s,3H).

Example 93: Preparation of 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylic acid

6-((4-((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylic acid

To a solution of methyl 6-((4-((6-amino-2,4-dimethylpyridin-3-yl)methylcarbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylate (50 mg, 0.1 mmol, 1 eq) in THF (10 mL) was added a solution of NaOH (4.9 mg, 0.12 mmol, 1.2 eq) in water (2 mL) and stirred at room temperature for 3 h. It was then acidified to pH 5 with AcOH. The mixture was concentrated in vacuo and purified by preparative HPLC to give 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylic acid (31.3 mg, 65.7%) as a gray solid. LRMS (M+H ⁺ ) m/z calcd 476.1, found 476.1.

¹ H NMR(DMSO-d6,400MHz)δ8.99(s,1H),8.74(s,1H),8.59-8.62(m,2H),8.18(s,1H),8.04(s,1H),7.79 (s,1H),7.60(d,1H),6.11(s,1H),5.64(s,2H),4.39(s,2H),4.33(d,2H),2.29(s,3H),2.16(s,3H).

Example 94: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-(hydroxymethyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-(hydroxymethyl)quinolin-6-yl)methyl)isonicotinamide

To a solution of methyl 6-((4-((6-amino-2,4-dimethylpyridin-3-yl)methylcarbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylate (100 mg, 0.2 mmol, ₁ equiv) in anhydrous THF was added LiAlH _dropwise at a temperature below -78 °C under N2 over 20 min. The reaction mixture was stirred for 5 h before being quenched with potassium sodium tartrate and extracted with EtOAc. The combined extracts were dried, concentrated, and the residue was purified by preparative HPLC to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-8-(hydroxymethyl)quinolin-6-yl)methyl)isonicotinamide (6.5 mg, 7%) as a white solid. LRMS (M+H ⁺ ) m/z calcd 462.2, found 462.1.

¹ H NMR (DMSO-d6, 400MHz) δ8.81 (d, 1H), 8.59-8.63 (m, 2H), 8.50 (d, 1H), 7.77 (s, 1H), 7.72 (d, 2H), 7.59 (d, 1H ), 6.13 (s, 1H), 5.72 (s, 2H), 5.26-5.28 (t, 1H), 5.07 (d, 1H), 4.32-4.33 (m, 4H), 2.30 (s, 3H), 2.16 (s, 3H).

Example 95: Preparation of methyl 6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylate

Methyl 6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylate

To a solution of 5-aminomethyl-6-methyl-pyridin-2-ylamine (89 mg, 0.43 mmol, 1.7 eq) in DMF (10 mL) was added 2-((3-chloro-8-(methoxycarbonyl)quinolin-6-yl)methyl)isonicotinic acid (90 mg, 0.25 mmol, 1 eq), HATU (123 mg, 0.32 mmol, _1.3 eq) and Et3N (0.5 mL). The mixture was stirred at room temperature overnight. It was then quenched with water and extracted with DCM. The combined extracts were dried and concentrated, and the residue was purified on a silica gel column (DCM/MeOH = 20/1, v/v) to give methyl 6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylate (100 mg, 84.7%) as a yellow solid. LRMS (M+H ^<+> ) m/z calcd. 476.1, found 476.1.

¹ H NMR(DMSO-d6,400MHz)δ8.98(t,1H),8.90(m,1H),8.61-8.63(m,2H),8.00(d,1H),7.92(d,1H),7.79(s,1H ),7.62(d,1H),7.25(d,1H),6.25(d,1H),5.81(s,2H),4.38(s,2H),4.28(d,2H),3.89(s,3H),2.28(s,3H).

Example 96: Preparation of 6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylic acid

6-((4-((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylic acid

6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylic acid (54 mg, 83.59%) was prepared as a white solid as described for 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxylic acid (Example 93). LRMS (M+H ⁺ ) m/z calcd 462.1, found 462.1.

¹ H NMR (DMSO-d6, 400MHz) δ9.07 (d, 1H), 8.99 (s, 1H), 8.83 (d, 1H), 8.62 (d, 1H), 8.41 (m, 1H), 8.17 (s, 1H), 7 .82(s, 1H), 7.62(d, 1H), 7.24(d, 2H), 6.21(d, 1H), 5.72(s, 1H), 4.45(s, 2H), 4.28(d, 2H), 2.27(s, 3H).

Example 97: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-8-(hydroxymethyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-8-(hydroxymethyl)quinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-8-(hydroxymethyl)quinolin-6-yl)methyl)isonicotinamide (Example 94) was prepared as a white solid (12.8 mg, 13.6%). LRMS (M+H ⁺ ) m/z calcd 448.1, found 448.1. ^1H NMR (DMSO-d6, 400MHz) δ8.98 (m, 1H), 8.80 (d, 1H), 8.62 (d, 1H), 8.50 (d, 1H), 7.72 (t, 3H), 7.62 (d, 1H), 7 .24(d,1H), 6.24(d,1H),5.75(s,2H),5.27(m,1H),5.08(d,2H),4.35(s,2H),4.27(d,1H),2.27(s,3H).

Example 98: Preparation of 6-amino-3-((2-((3-chloroquinolin-6-yl)methyl)isonicotinamido)methyl)-2,4-dimethylpyridine 1-oxide

6-amino-3-((2-((3-chloroquinolin-6-yl)methyl)isonicotinamido)methyl)-2,4-dimethylpyridine 1-oxide

To a solution of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide (50 mg, 0.11 mmol, 1 equiv) in anhydrous DCM was added m-CPBA. The reaction mixture was stirred for 3 h. It was then quenched with potassium sodium tartrate and extracted with DCM. The combined extracts were dried, concentrated, and the residue was purified by preparative HPLC to give 6-amino-3-((2-((3-chloroquinolin-6-yl)methyl)isonicotinamido)methyl)-2,4-dimethylpyridine 1-oxide (12.8 mg, 13.6%) as a white solid. LRMS (M+H ⁺ ) m/z calcd 448.1, found 448.1.

¹ H NMR(DMSO-d6,400MHz)δ8.82(s,1H),8.73(t,1H),8.60-8.61(d,1H),8.51(d,1H),7.96(d,1H),7.84(s ,1H),7.70-7.73(m,2H),7.57(d,1H),6.70(s,2H),6.53(s,1H),4.34(m,4H),2.41(s,3H),2.23(s,3H).

Example 99: Preparation of 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline 1-oxide

6-((4-((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline 1-oxide

Step 1: Preparation of 3-chloro-6-chloromethyl-quinoline 1-oxide

To a solution of 3-chloro-6-chloromethyl-quinoline (500 mg, 2.37 mmol, 1 equivalent) in anhydrous DCM (20 mL) was added m-CPBA (1.23 g, 7.11 mmol, 3 equivalents). The mixture was stirred at 40 ° C for 5 h. The reaction was then quenched with saturated NaHCO ₃ and extracted with DCM. The combined organic layers were dried and concentrated. The residue was purified by chromatography on a silica gel column (EtOAc/PE=1/3, v/v) to give 3-chloro-6-chloromethyl-quinoline 1-oxide (400 mg, 74%) as a yellow solid.

Step 2: Preparation of 3-chloro-6-((4-(methoxycarbonyl)pyridin-2-yl)methyl)quinoline 1-oxide

To a solution of 3-chloro-6-chloromethyl-quinoline 1-oxide (400 mg, 1.76 mmol, 1.0 equiv) in dioxane (10 mL) was added methyl 2-(trimethylstannyl)isonicotinate (583 mg, 1.94 mmol, 1.1 equiv) and Pd(PPh ₃ ) ₂ Cl ₂ (126 mg, 0.18 mmol, 0.1 equiv). The mixture was stirred at 90° C. for 3 h under a nitrogen atmosphere, then concentrated and purified by silica gel chromatography (DCM/MeOH=100/1, v/v) to give 3-chloro-6-((4-(methoxycarbonyl)pyridin-2-yl)methyl)quinoline 1-oxide (210 mg, 36%) as a yellow solid.

Step 3: Preparation of 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline 1-oxide

To a solution of 2-(3-chloro-1-oxo-quinolin-6-ylmethyl)-isonicotinic acid (100 mg, 0.32 mmol, 1 equivalent) in DMF (10 mL) was added 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine (106 mg, 0.48 mmol, 1.5 equivalents), HATU (182 mg, 0.48 mmol, 1.5 equivalents) and _Et N (1 mL). The mixture was stirred at room temperature for 3 h. It was then quenched with water and extracted with DCM. The combined extracts were dried, concentrated, and the residue was purified by preparative HPLC to give 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline 1-oxide (30 mg, 21%) as a gray solid. LRMS (M+H ⁺ ) m/z calcd. 448.1, found 447.8.

¹ H NMR(DMSO-d6,400MHz)δ8.74(d,1H),8.59-8.63(m,2H),8.38(d,1H),8.10(s,1H),7.91(s,1H),7.7 3-7.75(m,2H),7.59-7.61(m,1H),6.13(s,1H),5.71(s,2H),4.33(d,4H),2.30(s,3H),2.16(s,3H).

Example 100: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of 2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinic acid

To a solution of methyl 2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinate (300 mg, 0.86 mmol, 1 eq) in THF (10 mL)/water (5 mL) was added NaOH (42 mg, 1.05 mmol, 1.2 eq). The mixture was stirred at room temperature for 3 h, then aqueous HCl (2 N) was added to adjust the pH to 4. The mixture was extracted with EtOAc, and the organic layer was concentrated under pressure to provide the crude product (170 mg, 59%), which was not further purified.

Step 2: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(3,8-dichloro-quinolin-6-ylmethyl)-isonicotinic acid (80 mg, 0.24 mmol, 1 eq) in DMF (10 mL) was added 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine (83 mg, 0.36 mmol, 1.5 eq), HATU (110 mg, 0.29 mmol, 1.2 eq) and _Et3N (121.2 mg, 1.2 mmol, 5 eq). The mixture was stirred at room temperature for 3 h. It was then quenched with water and extracted with DCM. The combined extracts were dried, concentrated, and the residue was purified by preparative HPLC to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinamide (16 mg, 14%) as a white solid. LRMS (M+H ⁺ ) m/z calcd. 466.1, found 465.8.

¹ H NMR (DMSO-d6, 400MHz) δ8.95 (s, 1H), 8.60-8.66 (m, 3H), 7.77-7.94 (m, 3H), 8.00 (s, 1H), 7.60 (d , 1H), 7.68-7.71(m, 1H), 6.18(s, 1H), 5.91(s, 2H), 4.33-4.34(m, 4H), 2.32(s, 3H), 2.19(s, 3H).

Example 101: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinamide (25 mg, 23%) was prepared as a white solid as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3,8-dichloroquinolin-6-yl)methyl)isonicotinamide (Example 100). LRMS (M+H ⁺ ) m/z calcd 452.1, found 451.8.

¹ H NMR(DMSO-d6,400MHz)δ8.94-8.98(m,2H),8.63-8.64(m,2H),7.95(m,1H),7.84-7.8 8(m,2H),7.62(d,1H),6.24-6.26(m,1H),6.80(s,2H),4.28-4.36(m,4H),2.28(s,3H)

Example 102: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-5-fluoroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-5-fluoroquinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of 5-fluoroquinoline-6-carboxylic acid methyl ester and 7-fluoroquinoline-6-carboxylic acid methyl ester

To a suspension of 4-amino-2-fluorobenzoic acid methyl ester (20.0g, 0.129mol, 1 equivalent) and p-chloranil (35.0g, 0.142mol, 1.1 equivalents) in 500mL of 6N HCl solution, acrolein (13.5g, 0.194mol, 1.5 equivalents, 80% purity) is added. The mixture is stirred at 100°C for 10min. After cooling to room temperature, the mixture is alkalized to pH ₃ with saturated NaHCO. The precipitate is removed by filtration. The filtrate is extracted with _CHCl . The combined organic layer is dried and concentrated. The residue is purified by chromatography on a silica gel column (EtOAc/PE=1/10, v/v) to obtain 5-fluoroquinoline-6-methyl-formiate and 7-fluoroquinoline-6-methyl-formiate (3.0g, 11%) as a yellow solid.

Step 2: Preparation of methyl 3-chloro-5-fluoroquinoline-6-carboxylate and methyl 3-chloro-7-fluoroquinoline-6-carboxylate

To a solution of a mixture of 5-fluoroquinoline-6-methyl-formiate and 7-fluoroquinoline-6-methyl-formiate (3.7 g, 18.0 mmol, 1 equivalent) in DMF (90 mL) was added NCS (7.2 g, 54.0 mmol, 3 equivalents). The reaction mixture was stirred at 120 ° C for 40 min under N _2. The reaction mixture was cooled to ambient temperature, treated with water, neutralized with solid NaHCO ₃ , and stirred at room temperature for 30 min. Powdered sodium thiosulfate was carefully added to remove excess NCS. The mixture was extracted with EtOAc. The organic layer was dried and concentrated in vacuo. The crude product was purified by flash chromatography on a silica gel column to obtain a mixture of 3-chloro-5-fluoroquinoline-6-methyl-formiate and 3-chloro-7-fluoroquinoline-6-methyl-formiate (2.1 g, 49%) as a yellow solid.

Step 3: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-5-fluoroquinolin-6-yl)methyl)isonicotinamide

To a solution of methyl 3-chloro-5-fluoroquinoline-6-carboxylate (104.4 mg, 0.32 mmol, 1.0 equiv) in THF (5 mL)/ _H₂O (5 mL) was added _LiOH.H₂O (26.87 mg, 0.64 mmol, 2 equiv). The mixture was stirred at 40°C for 1 h and acidified to pH 5 with 1N HCl solution. The mixture was concentrated in vacuo, and the residue was used without further purification. To a solution of the above crude product in DMF (10 mL) were added 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine (106 mg, 0.48 mmol, 1.5 equiv), HATU (182 mg, 0.48 mmol, 1.5 equiv), and _Et₃N (1 mL). The mixture was stirred at room temperature for 3 h. It was then quenched with water and extracted with DCM. The combined extracts were dried and concentrated, and the residue was purified by preparative HPLC to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloro-5-fluoroquinolin-6-yl)methyl)isonicotinamide (17 mg, 12.4%) as a white solid. LRMS (M+H ⁺ ) m/z calcd 450.1, found 450.1. ¹ H NMR (DMSO-d6, 400 MHz) δ 8.87 (d, 1H), 8.55-8.58 (m, 3H), 7.93 (d, 1H), 7.73-7.81 (m, 2H), 7.60 (d, 1H), 6.11 (s, 1H), 5.66 (s, 2H), 4.33-4.38 (m, 4H), 2.29 (s, 3H), 2.16 (s, 3H).

Example 103: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-5-fluoroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-5-fluoroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloro-5-fluoroquinolin-6-yl)methyl)isonicotinamide (Example 102) was prepared as a white solid (45 mg, 32%). LRMS (M+H ⁺ ) m/z calcd 436.1, found 436.1. ^1H NMR(DMSO-d6,400MHz)δ8.99(m,1H),8.88(s,1H),8.57(d,2H),7.94(d,1H),7.63-7.81 (m,3H),7.24(s,1H),6.21(d,1H),5.72(s,2H),4.40(s,2H),4.27(d,2H),2.27(s,3H).

Example 104: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of methyl 3-chloroquinoline-6-carboxylate

To a solution of methyl quinoline-6-carboxylate (15.0 g, 80.2 mmol, 1.0 equiv) in DMF (200 ml) was added N-chlorosuccinimide (21.4 g, 0.16 mol, 2.0 equiv) and the reaction mixture was stirred at 120 ° C for 20 h. The reaction mixture was cooled to room temperature, treated with brine, and the mixture was extracted with EtOAc. The organic layer was dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by chromatography on silica gel (EtOAc/PE=1/8, v/v) to give methyl 3-chloroquinoline-6-carboxylate (9.1 g, 51%) as a yellow solid.

Step 2: Preparation of methyl (3-chloro-quinolin-6-yl)-methanol

To a solution of 3-chloroquinoline-6-formic acid methyl ester (8g, 36.0mmol, 1.0 equivalent) in anhydrous THF is added _LiAlH4 (2.5M in THF, 5.8mL, 0.4 equivalent). The resulting mixture is stirred at 0°C for 1h. Afterwards, additional _LiAlH4 (2.5M in THF, 2.8mL, 0.2 equivalent) is added. The system is stirred for another 30min at 0°C and quenched by slowly adding 1N aqueous NaOH. The resulting precipitate is filtered, and the filtrate is extracted with EtOAc. The combined organic layers are dried and concentrated. The residue is purified by silica gel chromatography (PE/EtOAc=20/1～5/1, v/v) to obtain (3-chloro-quinoline-6-yl)-methanol (4.8g, 69%) as a white solid.

Step 3: Preparation of 3-chloro-6-chloromethyl-quinoline

To (3-chloro-quinolin-6-yl)-methanol (3.3 g, 17.1 mmol, 1.0 equiv) was added _SOCl₂ (50 mL), and the mixture was stirred at room temperature for 1 h. The volatiles were removed under vacuum and the residue was dissolved in DCM. The mixture was washed with saturated aqueous _NaHCO₃ , dried, and concentrated to give 3-chloro-6-chloromethyl-quinoline (3.4 g, 94%) as a yellow solid.

Step 4: Preparation of methyl 2-methyl-6-(trimethylstannyl)isonicotinate

Hexamethyldistanane (0.21 mL, 334 mg, 1.02 mmol) and tetrakis(triphenylphosphine)palladium(0) (70 mg, 0.06 mmol) were added to a solution of methyl 2-chloro-6-methylisonicotinate (100 mg, 0.54 mmol) in anhydrous dioxane (10 mL), and the resulting mixture was refluxed under _N for 3 h. EtOAc (50 mL) and water (100 mL) were then added. The layers were separated and the organic layer was washed with water (5×100 mL), dried (Na ₂ SO ₄ ), and the solvent was removed by rotary evaporation to leave a crude residue that was used in the next step without further purification.

Step 4: Preparation of methyl 2-((3-chloroquinolin-6-yl)methyl)-6-methylisonicotinate

To a solution of 3-chloro-6-chloromethyl-quinoline (110 mg, 0.52 mmol, 1.0 eq) and crude methyl 2-methyl-6-(trimethylstannyl)isonicotinate in dioxane (10 mL) was added Pd( _PPh3 ) _2Cl2 ( ₃₆ mg, 0.05 mmol, 0.1 eq). The mixture was stirred at 90°C for 3 h under a nitrogen atmosphere, the solvent was removed, and the mixture was purified by silica gel chromatography (EtOAc/PE = 10/1 to 5:1, v/v) to give methyl 2-((3-chloroquinolin-6-yl)methyl)-6-methylisonicotinate (70 mg, 40%) as a yellow solid. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.83(d,1H),8.53(d,1H),7.98(d,1H),7.85(d,1H),7.73(dd,1H),7.58(s,1H),7.57(s,1H),4.36(s,2H),3.85(s,3H),2.53(s,3H).

Step 5: Preparation of 2-((3-chloroquinolin-6-yl)methyl)-6-methylisonicotinic acid

To a solution of 2-((3-chloroquinolin-6-yl)methyl)-6-methylisonicotinate (70 mg, 0.21 mmol, 1.0 equiv) in THF/ _H₂O (5 mL/1 mL) was added LiOH (71 mg, 2.1 mmol, 10 equiv). The resulting mixture was stirred at room temperature for 1 h; all starting material was consumed (estimated by TLC). The volatile solvents were removed on a rotary evaporator, and _the aqueous residue was neutralized with 1 M HCl and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine, dried over _Na₂SO₄ , and concentrated to provide the crude acid (50 mg, 75%), which was used directly in the next step without further purification.

Step 6: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)-6-methylisonicotinamide

To a solution of 2-((3-chloroquinolin-6-yl)methyl)-6-methylisonicotinic acid (50 mg, 0.16 mmol, 1.0 equiv) in DMF (5 mL) at 0°C was added 5-aminomethyl-6-methyl-pyridin-2-ylamine hydrochloride (33 mg, 0.0.19 mmol, 1.2 equiv), followed by HATU (91 mg, 0.24 mmol, 1.5 equiv) and DIPEA (0.08 mL, 0.48 mmol, 3.0 equiv). The reaction mixture was allowed to warm to room temperature and stirred under _N for 2 h. Water (20 mL) was added, and the mixture was extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine, dried over _Na2SO4 , _filtered , and concentrated. The residue was purified by preparative TLC (DCM:MeOH=15:1) to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloroquinolin-6-yl)methyl)-6-methylisonicotinoamide (24 mg, 34%) as a yellow solid. LRMS (M+H ⁺ ) m/z calcd 446.2, found 446.2. ¹ H NMR (DMSO-d ₆ ,400MHz): δ8.83(d,1H),8.66(s,1H),8.53(d,1H),7.97(d,1H),7.84(d,1H),7.71(dd,1H),7.50( s,1H),7.47(s,1H),6.32(s,2H),4.32(d,2H),4.29(s,2H),2.48(s,3H),2.37(s,3H),2.23(s,3H).

Example 105: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinamide

Step 1: Preparation of methyl 3-chloroquinoline-6-carboxylate

To a solution of methyl isonicotinate (5.0 g, 36.5 mmol, 1.0 equiv) in MeOH (70 ml) was added concentrated _{H₂SO₄ (} 300 mg, 3.1 mmol, 0.086 equiv) dropwise at room temperature. The mixture was heated under reflux, and _an aqueous solution _of ( _NH₄ ) _₂S₂O₈ (15.0 g, 65.7 mmol in 30 mL of water) was added dropwise. The reaction mixture was maintained at reflux for an additional 30 minutes, cooled to room temperature, and treated with 4M NaOH and _NaHCO₃ aqueous solution to approximately pH 7. The aqueous mixture was concentrated in vacuo, and the residue was extracted with EtOAc (100 mL x 2) _. The combined organic layers were washed with brine, dried over _Na₂SO₄ , filtered, and concentrated in vacuo. The crude product was purified by silica gel chromatography (PE/EtOAc = 1/3 to 1/1, v/v) to give methyl 3-chloroquinoline-6-carboxylate (1.5 g, 25%) as a white solid. LCMS (M+H ⁺ ) m/z calcd: 168, found: 168.0. ¹ H NMR (CDCl ₃ , 400 MHz): δ 8.71 (d, J = 4.8 Hz, 1 H), 7.84 (d, J = 0.8 Hz, 1 H), 7.77 (dd, 1 H), 4.84 (s, 2 H), 3.96 (s, 3 H).

Step 2: Preparation of methyl 2-(((methylsulfonyl)oxy)methyl)isonicotinate

To a stirred solution of methyl 2-(hydroxymethyl)isonicotinate (1.0 g, 6.0 mmol, 1.0 equiv) and TEA (1.2 g, 12.0 mmol, 2.0 equiv) in DCM (15 mL) was added MsCl (755 g, 6.6 mmol, 1.1 equiv) at 0° C. The resulting mixture was stirred at room temperature for another 30 minutes, diluted with DCM (60 mL), washed with water (30 mL), brine (30 mL×2), dried and concentrated to give methyl 2-(((methylsulfonyl)oxy)methyl)isonicotinate (1.2 g, 82%) as a dark brown oil.

Step 3: Preparation of methyl 2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinate

A mixture of methyl 2-(((methylsulfonyl)oxy)methyl)isonicotinate (300 mg, 1.22 mmol, 1.0 equiv), 5-chloro-1H-indazole (280 mg, 1.84 mmol, 1.5 equiv) and _K2CO3 (337 _mg , 2.44 mmol, 2 equiv) in DMF (5 mL) was stirred at 70°C for 2 hours. The mixture was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (30 mL), brine (30 mL x 2), dried and concentrated. The residue was purified by chromatography on silica gel (PE/EtOAc = 10/1 to 5/1 v/v) to give methyl 2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinate (120 mg, 33%) as a white solid. ¹ H NMR (CDCl ₃ , 400 MHz): δ 8.68 (d, 1H), 8.00 (s, 1H), 7.73 (d, 1H), 7.69 (d, 1H), 7.48 (s, 1H), 7.34 (d, 1H), 7.28 (dd, 1H), 5.73 (s, 2H), 3.85 (s, 3H). Chromatography on silica gel (PE/EtOAc=5/1 to 3/1, v/v) gave methyl 2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinate (70 mg, 19%) as a white solid. ¹ H NMR (CDCl ₃ , 400MHz): δ 8.73 (d, 1H), 8.05 (s, 1H), 7.79 (dd, 1H), 7.72 (s, 1H), 7.64-7.60 (m, 2H), 7.21 (dd, 1H), 5.75 (s, 2H), 3.90 (s, 3H).

Step 4: Preparation of 2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinic acid

To a solution of methyl 2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinate (270 mg, 0.89 mmol, 1.0 equiv) in THF (5 mL) was added LiOH.H ₂ O (375 mg, 8.9 mmol, 10.0 equiv) and water (5 mL). The mixture was stirred at room temperature for 2 hours and concentrated in vacuo to remove most of the THF. The aqueous mixture was adjusted to approximately pH 7 with 1 M HCl. The white suspension was filtered, and the solid was washed with water (10 mL) and evaporated to dryness in vacuo to give 2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinoic acid (240 mg, 93%) as a white solid.

Step 5: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinamide

To a stirred mixture of 2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinic acid (100 mg, 0.35 mmol, 1.0 equiv), TEA (101 mg, 1.0 mmol, 3.0 equiv) and 5-(aminomethyl)-4,6-dimethylpyridin-2-amine hydrochloride (65 mg, 0.35 mmol, 1.0 equiv) in DMF (3 mL) at 0° C. was added HATU (264 mg, 0.7 mmol, 2.0 equiv). The reaction mixture was stirred at room temperature for 16 h and then diluted with EtOAc (50 mL). The new mixture was washed with water (30 mL), brine (30 mL x 2), dried and concentrated. The residue was purified by chromatography on silica gel (DCM/MeOH=50/1 to 20/1, v/v) and then by preparative TLC (DCM/MeOH=20/1, v/v) to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinamide (10 mg, 7%) as a white solid. LCMS (M+H ⁺ ) m/z calcd 421, found 421.0.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 8.68 (t, 1H), 8.57 (d, 1H), 8.10 (d, 1H), 7.88 (d, 1H), 7.75 (d, 1H), 7.65 (dd, 1H), 7.49 (s, 1H), 7.40 (dd, 1H), 6.14 (s, 1H), 5.82-5.73 (m, 4H), 4.30 (d, 2H), 2.28 (s, 3H), 2.15 (s, 3H). LRMS (M+H ⁺ ) m/z calcd. 421.2, found 421.0.

Example 106: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinamide

Step 1: Preparation of 2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinic acid

To a solution of methyl 2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinate (150 mg, 0.50 mmol, 1.0 equiv) in THF (5 mL) was added LiOH H ₂ O (208 mg, 5.0 mmol, 10.0 equiv) and water (5 mL). The mixture was stirred at room temperature for 2 hours and concentrated in vacuo to remove most of the THF. The aqueous mixture was adjusted to pH ~7 with 1 M HCl. The white suspension was filtered, and the solid was washed with water (10 mL) and concentrated to give 2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinoic acid (110 mg, 77%) as a white solid.

Step 2: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinamide

To a stirred mixture of 2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinic acid (110 mg, 0.38 mmol, 1.0 equiv), TEA (105 mg, 1.0 mmol, 3.0 equiv) and 5-(aminomethyl)-4,6-dimethylpyridin-2-amine hydrochloride (71 mg, 0.38 mmol, 1.0 equiv) in DMF (3 mL) at 0° C. was added HATU (290 mg, 0.76 mmol, 2.0 equiv). The reaction mixture was stirred at room temperature for 16 h and then diluted with EtOAc (50 mL). The new mixture was washed with water (30 mL), brine (30 mL×2), dried and concentrated. The residue was purified by chromatography on silica gel (DCM/MeOH=50/1 to 10/1, v/v) and then by preparative TLC (DCM/MeOH=10/1, v/v) to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinamide (30 mg, 19%) as a white solid. LCMS (M+H ⁺ ) m/z calcd 421.2, found 421.0.

¹ H NMR (DMSO-d ₆ ,400MHz): δ8.84(s,1H),8.65(d,1H),8.55(d,1H),7.85(dd,1H),7.69(dd,1H),7.60-7.6 3(m,2H),7.23(dd,1H),6.38(s,1H),5.81(s,2H),4.33(d,2H),2.41(s,3H),2.27(s,3H).

Example 107: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-methyl-2H-indazol-2-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-methyl-2H-indazol-2-yl)methyl)isonicotinamide

Step 1: Preparation of methyl 2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinate

Methyl 2-((5-methyl-2H-indazol-2-yl)methyl)isonicotinate and methyl 2-((5-methyl-1H-indazol-1-yl)methyl)isonicotinate were prepared as described for methyl 2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinate.

¹ H NMR (CDCl ₃ ,400MHz): δ8.72(d,1H),7.98(s,1H),7.77(dd,1H),7.67(s,1H),7.61(d ,1H),7.39(s,1H),7.12(dd,1H),5.75(s,2H),3.88(s,3H),2.40(s,3H).

Methyl 2-((5-methyl-1H-indazol-1-yl)methyl)isonicotinate: ¹ H NMR (CDCl ₃ , 400 MHz): δ 8.72 (d, 1H), 8.00 (s, 1H), 7.72 (d, 1H), 7.51 (s, 1H), 7.47 (s, 1H), 7.29 (d, 1H), 7.18 (d, 1H), 5.76 (s, 2H), 3.84 (s, 3H), 2.43 (s, 3H).

Step 2: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinamide was prepared as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-2H-indazol-2-yl)methyl)isonicotinamide. LCMS (M+H+) m/z calcd 401.2, found 401.0. ¹ H NMR(DMSO-d6,400MHz): δ8.78(s,1H),8.63(d,1H),8.38(s,1H),7.75–7.64(m,1H),7.54(s,1H),7.47 (d,1H),7.46(s,1H),7.07(dd,1H),6.29(s,2H),5.75(s,2H),4.32(d,2H),2.35(s,6H),2.22(s,3H).

Example 108: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-methyl-3a,7a-dihydro-1H-indazol-1-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-methyl-3a,7a-dihydro-1H-indazol-1-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinamide was prepared as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((5-chloro-1H-indazol-1-yl)methyl)isonicotinamide (Example 105). LCMS (M+H ⁺ ) m/z calcd 401.2, found 401.0. ¹ H NMR (DMSO-d ₆ ,400MHz): δ8.68(t,1H),8.59(d,1H),8.01(d,1H),7.63(dd,1H),7.54(d,1H),7.53(s,1H),7.38(s,1H),7. 26–7.13(m,1H),6.16(s,1H),5.87(s,2H),5.75(s,2H),4.29(d,2H),2.40(s,3H),2.29(s,3H),2.15(s,3H).

Example 109: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-methyl-2-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide

N-((3-Chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-methyl-2-(methylsulfonyl)quinolin-3-yl)methyl)isonicotinamide (19 mg, 13% yield over 2 steps) was prepared as a purple solid as described for N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (Example 15). LRMS (M+H ⁺ ) m/z calcd 537.1, found 537.1. ¹ H NMR (DMSO-d ₆ ,400MHz)δ11.40(s,1H),9.27-9.24(t,1H),8.63-8.62(d,1H),8.31(s,1H),8.02-8.00(d,1H),7.82(s,1 H),7.75-7.45(m,5H),7.24-7.21(d,1H),4.78(s,2H),4.60-4.58(d,2H),3.52(s,3H),2.52-2.51(t,3H).

Example 110: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((2-methylquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((2-methylquinolin-6-yl)methyl)isonicotinamide (16 mg, 15%) was prepared as a yellow solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 459.1, found 459.1. ¹ H NMR (DMSO-d ₆ ,300MHz)δ11.39(s,1H),9.24(t,1H),8.66-8.64(d,1H),8.18-8.16(d,1H),7.85-7.78(m,3H),7.6 5-7.62(m,2H),7.50-7.36(m,3H),7.24-7.20(d,1H),4.59-4.57(d,2H),4.33(s,2H),2.62(s,3H).

Example 111: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-methylquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-methylquinolin-6-yl)methyl)isonicotinamide (30 mg, 20% yield over 2 steps) was prepared as described for N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 142). LRMS (M+H ⁺ ) m/z calcd 434.2, found 434.2. ¹ H NMR (CD ₃ OD,300MHz)δ8.64(d,1H),8.14(d,1H),8.06(d,1H),7.88(d,1H),7.77-7.59(m,6 H),7.47(d,1H),7.37(d,1H),6.90(d,1H),4.69(s,2H),4.38(s,2H),2.68(s,3H).

Example 112: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((2-methylquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((2-methylquinolin-6-yl)methyl)isonicotinamide (29 mg, 14%) was prepared as a yellow solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 398.1, found 398.1. ¹ H NMR (DMSO-d ₆ ,400MHz)δ9.02(t,1H),8.65-8.63(d,1H),8.18-8.16(d,1H),7.86-7.63(m,5H),7.38-7.36(d,2H),7 .26-7.24(d,1H),6.25-6.23(d,1H),5.77-5.76(d,2H),4.33-4.29(m,4H),2.63(s,3H),2.23(s,3H).

Example 113: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((2-methylquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((2-methylquinolin-6-yl)methyl)isonicotinamide (45 mg, 16%) was prepared as described in Step 7 of Example 24. LRMS (M+H ⁺ ) m/z calcd 411.9, found 411.9. ¹ H NMR (DMSO-d ₆ , 300 MHz) δ 8.59-8.65 (m, 2H), 8.17 (d, 1H), 7.83 (d, 1H), 7.75 (d, 2H), 7.58-7.64 (m, 2H), 7.37 (d, 1H), 6.11 (s, 1H), 5.67 (s, 2H), 4.30-4.34 (m, 3H), 2.61 (d, 3H), 2.29 (s, 3H), 2.15 (s, 3H).

Example 114: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide (29 mg, 21%) was prepared as a yellow solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 412.1, found 412.1. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 8.72 (s, 1H), 8.57-8.52 (d, 2H), 8.00 (s, 1H), 7.80-7.58 (m, 2H), 7.53-7.44 (m, 3H), 6.05 (s, 1H), 5.62 (s, 2H), 4.26 (s, 4H), 2.42-2.39 (m, 3H), 2.22 (s, 3H), 2.07 (s, 3H).

Example 115: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide (27 mg, 18% yield over 2 steps) was prepared as described for N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 142). LRMS (M+H ⁺ ) m/z calcd 434.2, found 434.2. ¹ H NMR (CD ₃ OD,400MHz)δ8.74(d,1H),8.66(d,1H),8.11(s,1H),8.07(d,1H),7.88(d,1H),7.83(s,1H),7.72(d,1H), 7.69(d,1H),7.61(s,2H),7.51(d,1H),7.49(d,1H),6.92(d,1H),4.71(s,2H),4.41(s,2H),2.51(s,3H).

Example 116: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide (62 mg, 40%) was prepared as an off-white solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 459, found 459. ¹ H NMR (DMSO-d ₆ ,400MHz)δ11.39(s,1H),9.24(t,1H),8.80(s,1H),8.64(d,1H),8.09(s,1H),7.87(d,1H),7.81(s,1H), 7.65-7.67(m,2H),7.50-7.55(m,2H),7.45(d,1H),7.23(d,1H),4.59(d,2H),4.37(s,2H),2.48(s,3H).

Example 117: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((6-methylquinolin-3-yl)methyl)isonicotinamide was prepared as described in Example 24, Step 7.

Example 118: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((6-fluoroquinolin-3-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((6-fluoroquinolin-3-yl)methyl)isonicotinamide (30 mg, 20% yield over 2 steps) was prepared as described for N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 142). LRMS (M+H ⁺ ) m/z calcd 438.2, found 438.2. ¹ H NMR (CD ₃ OD,300MHz)δ8.81(d,1H),8.66(d,1H),8.20(d,1H),8.08(d,1H),8.05(dd,1H),7.83(s,1H),7. 71(d,1H),7.70(d,1H),7.62(s,1H),7.57-7.47(m,3H),6.93(d,1H),4.72(s,2H),4.44(s,2H).

Example 119: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((6-fluoroquinolin-3-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((6-fluoroquinolin-3-yl)methyl)isonicotinamide (35 mg, 25%) was prepared as an off-white solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 416.1, found 416.1. ¹ H NMR (DMSO-d ₆ ,300MHz)δ8.87-8.86(d,1H),8.66-8.58(m,2H),8.18(d,1H),8.06-8.02(m,1H),7.78-7.72(m, 2H),7.64-7.58(m,2H),6.11(s,2H),5.68(s,2H),4.36-4.33(m,4H),2.30(s,3H),2.16(s,3H).

Example 120: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-fluoroquinolin-3-yl)methyl)isonicotinamide

N-((3-Chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-fluoroquinolin-3-yl)methyl)isonicotinamide (42 mg, 26.8%) was prepared as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 463.1, found 463.1. ¹ H NMR (DMSO-d ₆ ,300MHz)δ11.41(s,1H),9.26(t,1H),8.88(d,1H),8.65(d,1H),8.10(s,1H),8.02-8.07(m,1H),7.82(s ,1H),7.74(dd,1H),7.61-7.67(m,2H),7.51(s,1H),7.45(d,1H),7.22(d,1H),4.59(d,2H),4.39(s,2H).

Example 121: Preparation of N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((6-fluoroquinolin-3-yl)methyl)isonicotinamide

N-((6-amino-2-methylpyridin-3-yl)methyl)-2-((6-fluoroquinolin-3-yl)methyl)isonicotinamide (18 mg, 13.2%) was prepared as described in Step 7 of Example 24. LRMS (M+H ⁺ ) m/z calcd 401.9, found 401.9. ¹ H NMR (CD ₃ OD, 300 MHz) δ 8.80 (d, 1H), 8.63 (d, 1H), 8.20 (s, 1H), 8.01-8.04 (m, 1H), 7.78 (s, 1H), 7.63-7.64 (m, 1H), 7.52-7.58 (m, 3H), 7.40 (d, 1H), 4.43 (d, 4H), 2.38 (s, 3H).

Example 122: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((7-fluoroquinolin-3-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((7-fluoroquinolin-3-yl)methyl)isonicotinamide (35 mg, 25%) was prepared as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 415.9, found 415.9. ¹ H NMR (DMSO-d ₆ ,300MHz)δ8.92(d,1H),8.65(t,1H),8.60(d,1H),8.26(s,1H),8.04(dd,1H),7.79(s,1H),7.73(dd,1H) ,7.61(d,1H),7.51-7.55(m,1H),6.13(s,1H),5.68(s,2H),4.35-4.37(m,4H),2.31(s,3H),2.17(s,3H).

Example 123: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((7-fluoroquinolin-3-yl)methyl)isonicotinamide

N-((3-Chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((7-fluoroquinolin-3-yl)methyl)isonicotinamide (45 mg, 28.7%) was prepared as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 462.8, found 462.8. ¹ H NMR (DMSO-d ₆ ,300MHz)δ11.39-11.42(m,1H),9.26(t,1H),8.92(s,1H),8.64(d,1H),8.27(s,1H),8.04(dd, 1H),7.82(s,1H),7.65-7.74(m,2H),7.43-7.55(m,3H),7.22(d,1H),4.59(d,2H),4.38(s,2H).

Example 124: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((7-fluoroquinolin-3-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((7-fluoroquinolin-3-yl)methyl)isonicotinamide (16 mg, 10.8%) was prepared as described for N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 142). LRMS (M+H ⁺ ) m/z calcd 437.8, found 437.8. ¹ H NMR (CD ₃ OD,400MHz)δ8.72(d,1H),8.52(d,1H),8.09(s,1H),7.93(d,1H),7.77-7.80(m,1H),7.70(s,1H),7.56- 7.58(m,2H),7.47-7.50(m,2H),7.35(d,2H),7.26-7.31(m,1H),6.78(d,1H),4.58(s,2H),4.29(s,2H).

Example 125: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-(quinolin-3-ylmethyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-(quinolin-3-ylmethyl)isonicotinamide (24 mg, 17%) was prepared as a yellow solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 445.1, found 445.1. ¹ HNMR(DMSO-d ₆ ,400MHz)δ11.40(s,1H),9.26(t,1H),9.01(s,1H),8.66-8.65(d,1H),8.40(s,1H),8.05-8.00(t,2H),7.85( s,1H),7.80(t,1H),7.69-7.66(m,2H),7.51-7.44(m,2H),7.24-7.21(d,1H),4.60-4.59(d,2H),4.44(s,2H).

Example 126: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-chloroquinolin-3-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((6-chloroquinolin-3-yl)methyl)isonicotinamide (12 mg, 15%) was prepared as an off-white solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 479, found 479. ¹ H NMR (DMSO-d ₆ ,400MHz)δ11.39(s,1H),9.25(t,1H),8.92(d,1H),8.64(d,1H),8.20(s,1H),8.08(d,1H),8.00(d,1H), 7.82(s,1H),7.71(d,1H),7.66(d,2H),7.50(d,2H),7.45(d,1H),7.22(d,1H),4.59(d,2H),4.40(s,2H).

Example 127: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-methylquinolin-7-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-methylquinolin-7-yl)methyl)isonicotinamide (35 mg, 22%) was prepared as described for N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 142). LRMS (M+H ⁺ ) m/z calcd 434, found 434. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 9.39 (t, 1H), 8.67 (d, 1H), 8.18 (d, 1H), 8.13 (d, 1H), 7.80-7.85 (m, 3H), 7.75 (d, 1H), 7.68 (d, 1H), 7.54 (s, 1H), 7.47 (d, 1H), 7.35-7.40 (m, 2H), 6.84 (d, 1H), 6.72 (s, 2H), 4.60 (d, 2H), 4.37 (s, 2H), 2.62 (s, 3H).

Example 128: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((2-methylquinolin-7-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((2-methylquinolin-7-yl)methyl)isonicotinamide (25 mg, 15%) was prepared as described in Step 7 of Example 24. LRMS (M+H+) m/z calcd 459, found 459. ¹ H NMR (DMSO- _d6 , 300 MHz) δ 11.40 (br, 1H), 9.26 (t, 1H), 8.65 (d, 1H), 8.17 (d, 1H), 7.79-7.84 (m, 3H), 7.65 (d, 1H), 7.42-7.50 (m, 3H), 7.35 (d, 1H), 7.22 (d, 2H), 4.57 (d, 2H), 4.35 (s, 2H), 2.62 (s, 3H).

Example 129: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((2-methylquinolin-7-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((2-methylquinolin-7-yl)methyl)isonicotinamide (10 mg, 7%) was prepared as described in Step 7 of Example 24. LRMS (M+H ⁺ ) m/z calcd 412, found 412. ^1H NMR (DMSO- _d6 , 300 MHz) δ 8.59-8.64 (m, 2H), 8.17 (d, 1H), 7.82 (d, 1H), 7.76 (d, 2H), 7.60 (d, 1H), 7.45 (d, 1H), 7.35 (d, 1H), 6.10 (s, 1H), 5.67 (s, 2H), 4.33 (s, 4H), 2.62 (s, 3H), 2.29 (s, 3H), 2.15 (s, 3H).

Example 130: Preparation of N-(3-chloro-6-fluoro-1H-indol-5-ylmethyl)-2-(2-cyano-quinolin-6-ylmethyl)-isonicotinamide

N-(3-Chloro-6-fluoro-1H-indol-5-ylmethyl)-2-(2-cyano-quinolin-6-ylmethyl)-isonicotinamide (90 mg, 22%) was prepared as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 570.1, found 570.1. ¹ H NMR (DMSO-d ₆ ,400MHz)δ11.41(s,1H),9.26(t,1H),8.60-8.65(m,2H),8.00-8.08(m,3H),7.87-7.89(dd,1H),7.81(s,1H), 7.66-7.67(dd,1H),7.50-7.51(d,1H),7.43-7.45(d,1H),7.21-7.24(d,1H),4.58-4.59(d,2H),4.42(s,2H).

Example 131: Preparation of N-(6-amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(2-cyano-quinolin-6-ylmethyl)-isonicotinamide

N-(6-Amino-2,4-dimethyl-pyridin-3-ylmethyl)-2-(2-cyano-quinolin-6-ylmethyl)-isonicotinamide (90 mg, 26%) was prepared as described in Step 7 of Example 24. LRMS (M+H ⁺ ) m/z calcd 423.1, found 423.1. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 8.60-8.64 (m, 3H), 8.00-8.07 (m, 3H), 7.86-7.88 (m, 1H), 7.78 (s, 1H), 7.61-7.62 (d, 1H), 6.12 (s, 1H), 5.68 (s, 2H), 4.33-4.40 (m, 4H), 2.30 (s, 3H), 2.16 (s, 3H).

Example 132: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylisoquinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of 6-bromo-3-methyl-isoquinoline

To a solution of 4-bromo-benzylamine (10.0 g, 54 mmol, 1.0 equivalent) in DCE (100 mL) was added 1,1-dimethoxy-propan-2-one (7.0 g, 59 mmol, 1.1 equivalents) and MgSO ₄ (20 g). The mixture was stirred at 40 ° C overnight. NaBH ₃ CN (4.08 g, 64.8 mmol, 1.2 equivalents) was then added to the mixture. After stirring at room temperature for 5 h, the mixture was filtered. The filtrate was concentrated to obtain a yellow oil. Chlorosulfonic acid (30 mL) was cooled to -10 ° C, and the above-mentioned crude product was added dropwise. The reaction mixture was heated to 100 ° C for 10 min, then cooled and poured into ice. The mixture was neutralized with 2M NaOH and extracted with EtOAc. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=2/1, v/v) to give 6-bromo-3-methyl-isoquinoline (4.0 g, 34% yield over 3 steps) as a yellow solid.

Step 2: Preparation of methyl 3-methylisoquinoline-6-carboxylate

To a high pressure vessel was added 6-bromo-3-methyl-isoquinoline (4.0 g, 18 mmol), Pd(dppf)Cl ₂ (735 mg, 0.9 mmol, 0.05 eq) and triethylamine (5.0 mL, 36 mmol, 2 eq) in 40 mL of methanol. The vessel was purged three times with nitrogen and three times with carbon monoxide. The vessel was pressurized to 3 MPa with carbon monoxide and heated to 100° C. The reaction was then stirred overnight before being allowed to cool to room temperature. The resulting solution was concentrated and purified by flash chromatography on silica gel (PE/EtOAc=1/1, v/v) to give 3-methyl-isoquinoline-6-carboxylic acid methyl ester (3.4 g, 94%) as a white solid.

Step 3: Preparation of (3-methyl-isoquinolin-6-yl)-methanol

To a solution of 3-methyl-isoquinoline-6-carboxylic acid methyl ester (3.3 g, 16.42 mmol, 1 eq) in anhydrous THF (100 mL) was added LiAlH(t-BuO) ₃ (12.5 g, 45.25 mmol, 3 eq). The resulting mixture was stirred at 60 ° C for 5 h and then quenched by adding water. The mixture was extracted with EtOAc. The combined extracts were dried and concentrated. The residue was purified by silica gel chromatography (PE/EtOAc=1/1, v/v) to give (3-methyl-isoquinolin-6-yl)-methanol (2.5 g, 89%) as a white solid

Step 4: Preparation of 6-chloromethyl-3-methyl-isoquinoline

To (3-methyl-isoquinolin-6-yl)-methanol (1.5 g, 8.67 mmol, 1 eq) was added _SOCl₂ (9 mL), and the mixture was stirred at room temperature for 3 h. The volatiles were then removed under vacuum at 40° C., and the residue was dissolved in DCM. The mixture was washed with saturated aqueous _NaHCO₃ , dried, and concentrated to give 6-chloromethyl-3-methyl-isoquinoline (1.4 g, 85%) as a white solid.

Step 5: Preparation of methyl 2-((3-methylisoquinolin-6-yl)methyl)isonicotinate

Methyl 2-((3-methylisoquinolin-6-yl)methyl)isonicotinate (1.0 g, 47%) was prepared as described for Example 24, Step 5.

Step 6: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylisoquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylisoquinolin-6-yl)methyl)isonicotinamide (20 mg, 14% yield over 2 steps) was prepared as described for N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 142). LRMS (M+H ⁺ ) m/z calcd 434.2, found 434.2. ¹ H NMR (CD ₃ OD,400MHz)δ9.06(s,1H),8.66(d,1H),8.08(d,1H),7.97(d,1H),7.80(s,1H),7.71(s,2H),7.70(s,1H), 7.62(s,1H),7.56(s,1H),7.50(d,1H),7.47(d,1H),6.92(d,1H),4.71(s,2H),4.11(s,2H),2.64(s,3H).

Example 133: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-methylisoquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-methylisoquinolin-6-yl)methyl)isonicotinamide (43 mg, 28% yield over 2 steps) was prepared as described for Example 132. LRMS (M+H ⁺ ) m/z calcd 459.1, found 459.1. ¹ H NMR (DMSO-d ₆ ,400MHz)δ11.42(s,1H),9.26(t,1H),9.15(s,1H),8.67(d,1H),8.00(dd,1H),7.79(s,1H),7.72(s,1H),7.67( d,1H),7.58(s,1H),7.53(d,1H),7.52(s,1H),7.46(d,1H),7.24(d,1H),4.60(d,2H),4.36(s,2H),2.59(s,3H).

Example 134: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (34 mg, 34% yield over 2 steps) was prepared as described for N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 142). LRMS (M+H ⁺ ) m/z calcd 498.2, found 498.2. ¹ H NMR (CD ₃ OD,400MHz)δ8.66(d,1H),8.49(d,1H),8.06(d,1H),8.03(d,1H),8.02(s,1H),7.88(s,1H),7.81(s,1H),7.78( d,1H),7.69(d,1H),7.67(d,1H),7.59(s,1H),7.46(d,1H),6.88(d,1H),4.69(s,2H),4.43(s,2H),3.37(s,3H).

Example 135: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide

N-((3-Chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((2-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide (24 mg, 23%) was prepared as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 523.1, found 523.1. ¹ H NMR (CD ₃ OD,400MHz)δ8.53(d,1H),8.47(d,1H),8.02(d,1H),7.99(d,1H),7.84(s,1H),7.75(d,1H),7.68( s,1H),7.56(d,1H),7.40(d,1H),7.13(s,1H),7.02(s,1H),4.58(s,2H),4.36(s,2H),3.25(s,3H).

Example 136: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((4-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((4-cyanoquinolin-6-yl)methyl)isonicotinamide (43 mg, 29% yield over 2 steps) was prepared as described for N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 142). LRMS (M+H ⁺ ) m/z calcd 445.2, found 445.2. ¹ H NMR (CD ₃ OD,300MHz)δ9.40(t,1H),9.06(d,1H),8.69(d,1H),8.15-8.11(m,3H),8.06(s,1H),7.92(d,1H),7.87(s,1 H),7.77(d,1H),7.71(d,1H),7.56(s,1H),7.41(d,1H),6.85(d,1H),6.73(s,2H),4.62(d,2H),4.50(s,2H).

Example 137: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((4-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((4-cyanoquinolin-6-yl)methyl)isonicotinamide (45 mg, 29%) was prepared as a yellow solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 470.1, found 470.1. ¹ H NMR(DMSO,400MHz)δ11.42(s,1H),9.28(t,1H),9.07(d,1H),8.68(d,1H),8.14(d,1H),8.12(s,1H),8.06(s, 1H),7.91(d,1H),7.86(s,1H),7.68(d,1H),7.51(d,1H),7.46(d,1H),7.24(d,1H),4.60(d,1H),4.49(s,2H).

Example 138: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((4-cyanoquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((4-cyanoquinolin-6-yl)methyl)isonicotinamide (23 mg, 17% yield) was prepared as a yellow solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 423.2, found 423.2. ¹ H NMR (CD ₃ OD, 300 MHz) δ 8.98 (d, 1H), 8.63 (d, 1H), 8.10 (d, 1H), 8.07 (s, 1H), 7.93 (d, 1H), 7.86 (dd, 1H), 7.77 (s, 1H), 7.63 (dd, 1H), 6.29 (s, 1H), 4.50 (s, 2H), 4.49 (s, 2H), 2.38 (s, 3H), 2.26 (s, 3H).

Example 139: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((7-chloroquinolin-3-yl)methyl)isonicotinamide

N-((1-aminoisoquinolin-6-yl)methyl)-2-((7-chloroquinolin-3-yl)methyl)isonicotinamide (34 mg, 23% yield over 2 steps) was prepared as described for N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (Example 142). LRMS (M+H ⁺ ) m/z calcd 454.1, found 454.1. ¹ H NMR (DMSO-d ₆ ,400MHz)δ9.42(d,1H),8.96(s,1H),8.68(d,1H),8.20(s,1H),8.15(d,1H),8.05(s,1H),8.02(d,1H),7.86(s, 1H),7.78(d,1H),7.70(d,1H),7.63(dd,1H),7.56(s,1H),7.42(d,1H),6.86(d,1H),4.63(d,2H),4.42(s,2H).

Example 140: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((7-chloroquinolin-3-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((7-chloroquinolin-3-yl)methyl)isonicotinamide (35 mg, 23%) was prepared as a white solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 479.1, found 479.1. ¹ H NMR (DMSO-d ₆ ,400MHz)δ11.42(s,1H),9.27(t,1H),8.95(d,1H),8.66(d,1H),8.27(s,1H),8.04(d,1H),8.01(d,1H),7 .83(s,1H),7.67(d,1H),7.62(dd,1H),7.52(d,1H),7.46(d,1H),7.25(d,1H),4.60(d,2H),4.40(s,2H).

Example 141: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((7-chloroquinolin-3-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((7-chloroquinolin-3-yl)methyl)isonicotinamide (10 mg, 7%) was prepared as a white solid as described in Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 432.2, found 432.2. ¹ H NMR (CD ₃ OD, 400 MHz) δ 8.87 (d, 1H), 8.63 (d, 1H), 8.25 (s, 1H), 8.00 (s, 1H), 7.91 (d, 1H), 7.78 (s, 1H), 7.63 (d, 1H), 7.60 (dd, 1H), 6.32 (s, 1H), 4.53 (d, 2H), 4.44 (s, 2H), 2.41 (s, 3H), 2.29 (s, 3H).

Example 142: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(3-methyl-quinolin-6-ylmethyl)-isonicotinic acid (150 mg, crude) in DMF (15 mL) was added 6-aminomethyl-isoquinolin-1-ylamine (62 mg, 0.36 mmol, 1.0 equiv), followed by EDCI (104 mg, 0.54 mmol, 1.5 equiv), HOBT (73 mg, 0.54 mmol, 1.5 equiv), and TEA (109 mg, 1.08 mmol, 3.0 equiv). The reaction mixture was heated to 40° C. and stirred overnight. Water was added, and the mixture was extracted with DCM. The organic layer was washed with water, dried over Na ₂ SO ₄ , filtered, and concentrated. The residue was purified by preparative HPLC to give N-((1-aminoisoquinolin-6-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isoamide (30 mg, 19%) as a white solid. LRMS (M+H ⁺ ) m/z calcd. 434, found 434. ¹ H NMR (DMSO-d ₆ , 400 MHz) δ 9.40 (t, 1H), 8.71 (s, 1H), 8.67 (d, 1H), 8.13 (d, 1H), 8.07 (s, 1H), 7.90 (d, 1H), 7.75-7.80 (m, 3H), 7.68 (d, 1H), 7.62 (d, 1H), 7.54 (s, 1H), 7.49 (d, 1H), 6.84 (d, 1H), 6.77 (s, 2H), 4.60 (d, 2H), 4.35 (s, 2H), 2.46 (s, 3H).

Example 143: Preparation of N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide

N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (30 mg, 18%) was prepared as a yellow solid as described for Example 24, Step 7. LRMS (M+H+) m/z calcd 459, found 459. ¹ H NMR (DMSO-d ₆ ,300MHz)δ11.42(br,1H),9.25(t,1H),8.71(s,1H),8.65(d,1H),8.05(s,1H),7.89(d,1H),7.76(d,2 H),7.60-7.66(m,2H),7.51(d,1H),7.43(d,1H),7.22(d,2H),4.58(d,2H),4.34(s,2H),2.46(s,3H).

Example 144: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide

To a solution of 2-(3-methyl-quinolin-6-ylmethyl)-isonicotinic acid (150 mg, crude) in DMF (15 mL) was added 5-aminomethyl-4,6-dimethyl-pyridin-2-ylamine dihydrochloride (400 mg, crude), followed by EDCI (104 mg, 0.54 mmol, 1.5 eq), HOBT (73 mg, 0.54 mmol, 1.5 eq) and TEA (109 mg, 1.08 mmol, 3.0 eq). The reaction mixture was heated to 40 ° C and stirred overnight. Water was added and the mixture was extracted with DCM. The organic layer was washed with water, dried over Na ₂ SO ₄ , filtered, and concentrated. The residue was purified by preparative HPLC to give N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methylquinolin-6-yl)methyl)isonicotinamide (25 mg, 17%) as a yellow solid. LRMS (M+H ⁺ ) m/z calcd. 412, found 412. ¹ H NMR (DMSO-d ₆ , 300 MHz) δ 8.71 (s, 1H), 8.65 (t, 1H), 8.60 (d, 1H), 8.06 (s, 1H), 7.89 (d, 1H), 7.74 (d, 2H), 7.60 (d, 2H), 6.10 (s, 1H), 5.70 (s, 2H), 4.32 (s, 4H), 2.46 (s, 3H), 2.29 (s, 3H), 2.15 (s, 3H).

Example 145: Preparation of N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methylisoquinolin-6-yl)methyl)isonicotinamide

N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methylisoquinolin-6-yl)methyl)isonicotinamide (29 mg, 21%) was prepared as described for Example 24, Step 7. LRMS (M+H ⁺ ) m/z calcd 412.2, found 412.2. ¹ H NMR (CD ₃ OD,400MHz)δ9.05(s,1H),8.61(d,1H),7.96(s,1H),7.73(s,1H),7.69(s,1H),7.62(dd,1H),7.55( s,1H),7.52(d,1H),6.30(s,1H),4.49(s,2H),4.38(s,2H),2.64(s,3H),2.39(s,3H),2.26(s,3H).

Example 146: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-(aminomethyl)quinolin-6-yl)methyl)isonicotinamide

Step 1: Preparation of tert-butyl (6-((4-((1-aminoisoquinolin-6-yl)methylcarbamoyl)pyridin-2-yl)methyl)quinolin-2-yl)methylcarbamate

tert-Butyl (6-((4-((l-aminoisoquinolin-6-yl)methylcarbamoyl)pyridin-2-yl)methyl)quinolin-2-yl)methylcarbamate (70 mg, 35%) was prepared as described for Example 24, Step 7.

Step 2: Preparation of N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-(aminomethyl)quinolin-6-yl)methyl)isonicotinamide

To a solution of tert-butyl (6-((4-((1-aminoisoquinolin-6-yl)methylcarbamoyl)pyridin-2-yl)methyl)quinolin-2-yl)methylcarbamate (80 mg, 0.13 mmol) in EtOAc (1 mL) was added HCl/EtOAc solution. The mixture was stirred at room temperature for 1 h. The precipitate was collected by filtration to give N-((1-aminoisoquinolin-6-yl)methyl)-2-((2-(aminomethyl)quinolin-6-yl)methyl)isonicotinamide (50 mg, 66%) as an off-white solid. LRMS (M+ ^H ) m/z calcd 449.2, found 449.2. ¹ H NMR (DMSO-d ₆ ,400MHz)δ13.51(s,1H),10.05(s,1H),9.25(br,1H),8.88(d,1H),8.63-8.62(m,4H),8.45(d,1H),8.19(s,1H),8. 09-8.01(m,3H),7.89(d,1H),7.85(s,1H),7.73-7.64(m,4H),7.20(d,1H),4.69(d,2H),4.59(s,2H),4.39(t,2H).

II. Biological Evaluation

Example 1: In vitro enzyme inhibition

The ability of compounds disclosed herein to inhibit human plasma kallikrein activity was quantified according to the following procedure.

Prepare a 10 mM solution of the test compound in DMSO. This solution is serially diluted 1:5 in DMSO to produce 2000, 400, 80, 16, 3.2, 0.64, 0.128, 0.0256, and 0.00512 μM compound test solutions. Include a control tube containing only DMSO. 16 μL of each compound test solution is combined with 384 μL of assay buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.01% Triton X-100) to create a "4X test compound buffer stock."

In addition, a 40 nM solution of human plasma kallikrein (Abeam) and a 93.6 μM solution of Pro-Phe-Arg-AMC (Bachem) were prepared using assay buffer. These solutions are referred to herein as 4X hPK and 2X PFR-AMC, respectively.

60 μL of each 4X test compound buffer stock solution was combined with 60 μL of 4X hPK to produce 120 μL of "2X test compound buffer stock solution/2X hPK". 50 μL was taken out of this mixture and placed in duplicate wells on a Microfluor 1 Black U-bottom microtiter plate (Thermo Scientific). The plate was incubated at 37°C for 5 minutes. 50 μL of pre-warmed 2X PFR-AMC was added to each well to start the enzymatic reaction. The cleavage of PFR-AMC was monitored in a Biotek Synergy H4 reader set at 37°C. Readings were taken every 43 seconds for 1 hour. The highest average speed among 20 reads (approximately 15 minutes) was used to calculate IC _50. IC ₅₀ was calculated using Gen5 (Biotek Instruments).

The compounds in Table 3 were tested for their ability to inhibit human plasma kallikrein activity.

Table 3

Note: _IC50 data for biochemical assays are given in the following ranges:

A:≤0.10μM C:>1.0μM to ≤10μM

B:>0.10μM to ≤1.0μM D:>10μM

Example 2: In vitro cell assay

The ability of the compounds disclosed herein to inhibit cellular kallikrein activity was quantified and respective _EC50 values were determined.

Material:

Plasma kallikrein inhibitor C1NH (Athens Research & Technology, Cat#16-16-031509); ellagic acid (Sigma, E2250); substrate Z-FR-2-AMC (GL Biochem, Cat#55352); Nunc™ 96-well polypropylene MicroWell™ plates (Nunc, Cat#267342)

method:

All dilutions were prepared in assay buffer containing 50 mM Tris-HCl pH 7.2, 150 mM NaCl, and 0.01% Triton X-100.

Four-fold serial dilutions were prepared from a 107.53 μM stock solution of the plasma kallikrein inhibitor C1NH to yield 10 solutions with concentrations ranging from 20 μM to 0.76 nM. Similarly, four-fold serial dilutions were prepared from a 10 mM stock solution of each test compound to yield 10 solutions with concentrations ranging from 4 mM to 0.015 μM. The 10 solutions of test compounds prepared by serial dilution were further diluted 50-fold in assay buffer.

Human plasma was thawed on ice and centrifuged at 4°C for 15 minutes to remove platelets. A 1 mM stock solution of ellagic acid was diluted to 8 μM and mixed with platelet-depleted human plasma at a ratio of 1:0.8. The mixture of human plasma and ellagic acid was further diluted 32-fold in assay buffer to produce the final mixture for inhibition testing.

A volume of 22.5 μL of the final mixture of human plasma and ellagic acid was added to a 96-well microplate, and the plate was incubated at 37°C for 15 min.

To the inhibitor control wells, different concentrations of CINH inhibitors prepared by serial dilution as described above were added. The volume of CINH inhibitor added to the inhibitor control wells was 12.5 μL, resulting in final concentrations of 5 μM, 1.25 μM, 312.5 nM, 78.125 nM, 19.531 nM, 4.883 nM, 1.221 nM, 0.305 nM, 0.076 nM, and 0.019 nM. Each CINH concentration was tested in duplicate.

Different concentrations of test compound, also prepared by serial dilution as described above, were added to the test wells. The volume of test compound added to each test well was 12.5 μL, resulting in final concentrations of 20 μM, 5 μM, 1.25 μM, 312.5 nM, 78.125 nM, 19.531 nM, 4.883 nM, 1.221 nM, 0.305 nM, and 0.076 nM. Each test compound concentration was tested in duplicate.

In addition to the inhibitor control and test wells, the 96-well assay plate also includes positive control wells containing a mixture of human plasma and ellagic acid without the C1NH inhibitor or test compound, and background wells containing neither the mixture of human plasma and ellagic acid nor the test compound. The total volume of the liquid in the positive control and background wells is brought to 35 μL with assay buffer.

Assay plates containing C1NH inhibitors or test compounds mixed with human plasma and ellagic acid, as well as appropriate controls, were incubated at 37°C for 5 minutes. A 10 mM stock solution of the substrate, Z-FR-2-AMC, was diluted to 133.2 μM in assay buffer, and 15 μL of the diluted substrate was added to each well, yielding a final substrate concentration of 40 μM in each well. The reagents were mixed by gently shaking the plate for 30 seconds.

The enzyme reaction was quantified by direct kinetic reading of the assay plate using excitation/emission wavelengths of 330 nm/440 nm, respectively. Fluorescence intensity was recorded for 60 min at 43 sec intervals.

The inhibitory activity of the test compounds was estimated using IC50 values, calculated from the dose-response curves of the test compounds, and fitted with the “log (inhibitor)-response (variable slope)” equation in GraphPad Prism software (GraphPad Software, Inc.).

The percentage of inhibition was calculated using the following equation:

Wherein, mean (BG) is the mean value of the fluorescence intensity of the background wells, and mean (PC) is the mean value of the fluorescence intensity of the positive control wells.

Table 4 provides _EC50 values for various compounds disclosed herein.

Table 4

Note: Experimental _IC50 data are given in the following ranges:

A:≤0.10μM C:>1.0μM to ≤10μM

B:>0.10μM to ≤1.0μM D:>10μM

III. Preparation of pharmaceutical dosage forms

Example 1: Oral tablets

Tablets are prepared by mixing 48% by weight of a compound of formula (I) or a pharmaceutically acceptable salt thereof, 45% by weight of microcrystalline cellulose, 5% by weight of low-substituted hydroxypropyl cellulose, and 2% by weight of magnesium stearate. Tablets are prepared by direct compression. The total weight of the compressed tablets is maintained at 250-500 mg.

Claims (17)

1. A compound, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide; N-((1-Aminoisoquinoline-6-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide; N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide; N-((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide; N-((5-chloro-1H-indazol-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide; N-((3-chloro-4-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinolin-6-yl)methyl)isonicotinamide; N-((1-aminoisoquinoline-6-yl)methyl)-2-((3-chloroquinoline-6-yl)methyl)isonicotinamide; N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloroquinoline-6-yl)methyl)isonicotinamide; 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide; and 6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide. 2. A pharmaceutical composition comprising the compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 3. Use of the compound of claim 1 in the preparation of compositions for inhibiting kallikrein. 4. Use of the compound of claim 1 in the preparation of a medicament for treating angioedema in patients in need. 5. The compound of claim 1, wherein the compound is N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide or a pharmaceutically acceptable salt thereof. 6. The compound of claim 1, wherein the compound is N-((1-aminoisoquinoline-6-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide or a pharmaceutically acceptable salt thereof. 7. The compound of claim 1, wherein the compound is N-((3-chloro-6-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide or a pharmaceutically acceptable salt thereof. 8. The compound of claim 1, wherein the compound is N-((3-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide or a pharmaceutically acceptable salt thereof. 9. The compound of claim 1, wherein the compound is N-((5-chloro-1H-indazol-3-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide or a pharmaceutically acceptable salt thereof. 10. The compound of claim 1, wherein the compound is N-((3-chloro-4-fluoro-1H-indol-5-yl)methyl)-2-((3-methyl-8-(methylsulfonyl)quinoline-6-yl)methyl)isonicotinamide or a pharmaceutically acceptable salt thereof. 11. The compound of claim 1, wherein the compound is N-((1-aminoisoquinoline-6-yl)methyl)-2-((3-chloroquinoline-6-yl)methyl)isonicotinamide or a pharmaceutically acceptable salt thereof. 12. The compound of claim 1, wherein the compound is N-((6-amino-2,4-dimethylpyridin-3-yl)methyl)-2-((3-chloroquinoline-6-yl)methyl)isonicotinamide or a pharmaceutically acceptable salt thereof. 13. The compound of claim 1, wherein the compound is 6-((4-(((6-amino-2,4-dimethylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide or a pharmaceutically acceptable salt thereof. 14. The compound of claim 1, wherein the compound is 6-((4-(((6-amino-2-methylpyridin-3-yl)methyl)carbamoyl)pyridin-2-yl)methyl)-3-chloroquinoline-8-carboxamide or a pharmaceutically acceptable salt thereof. 15. A pharmaceutical composition comprising a compound as claimed in any one of claims 5 to 14, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 16. Use of the compound according to any one of claims 5 to 14 in the preparation of a composition for inhibiting kallikrein. 17. Use of the compound according to any one of claims 5 to 14 in the preparation of a medicament for treating angioedema in patients in need.