Inhibitors of topoisomerase i

Abstract

Disclosed herein are topoisomerase I compounds represented by Formula I, or a pharmaceutically acceptable salt thereof, and methods of treating proliferative disorders in a subject, comprising administering a compound disclosed herein:

Description

[0001] Attorney Docket No.: LCH-03225

[0002] INHIBITORS OF TOPOISOMERASE

[0003] RELATED APPLICATIONS

[0004] This application claims the benefit of U.S. Provisional Application No. 63 / 725,138, filed November 26, 2024, the contents of which is hereby incorporated by reference herein.

[0005] BACKGROUND

[0006] DNA topoisomerases are enzymes that create irreversible cuts in one of the two strands of double-stranded DNA. Topoisomerases catalyze changes in the topological state of DNA, and are crucial for the successful replication, transcription and recombination of DNA. As a result, topoisomerases are potential targets for anticancer therapy. Topoisomerases are categorized into two major subtypes: topoisomerase I and topoisomerase II. Topoisomerase I (TOPI) belongs to a family of ATP-independent enzymes that resolve topological constraints associated with DNA supercoils during DNA transcription and replication, chromosome condensation, and intertwined DNA during mitosis. Topoisomerase I relieves the torsional DNA strain by creating a single strand nick in the phosphodiester backbone, covalently ligating the enzyme to DNA until the strain is relieved J. Med. Chem. 2007, 50, 18, 4419-4430). TOPI is known to be inhibited by camptothecin, the first developed and one of the most investigated topoisomerase inhibitors. However, camptothecin inhibitors are associated with low pharmacokinetic efficiency and low stability due to the hydrolysis of the lactone ring, which results in hydroxy acid as a byproduct, and has a high affinity for human serum albumin Biomolecules 2015; 5(3): 1652-1670).

[0007] Therefore, there is a continuing need to discover and develop novel and potent topoisomerase inhibitors as therapeutic agents for cancer and various proliferative disorders.

[0008] SUMMARY OF THE INVENTION

[0009] In certain aspects, the present disclosure provides indenoisoquinoline-based, non- camptothecin compounds. In certain aspects, the present disclosure provides compounds having a structure represented by Formula I, or a pharmaceutically acceptable salt thereof:

[0010] -1-

[0011] FH13103096.12 Attorney Docket No.: LCH-03225

[0012] I wherein

[0013] R1, R2, R3and R4are each independently H, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, cycloalkyl, or heterocyclyl; or R2and R3, together with the carbon atoms that separate them, complete a heterocyclyl;

[0014] Y is alkylene, alkenylene, cycloalkylene, heteroalkylene, or heteroalkenylene;

[0015] Z is aryl, heteroaryl, cycloalkyl, heterocyclyl, amido, or amino; and

[0016] A is aryl or heteroaryl.

[0017] In other aspects, the present disclosure provides methods of inhibiting topoisomerase I in a subject, comprising administering a compound disclosed herein, or a pharmaceutically acceptable salt thereof, to the subject.

[0018] In certain aspects, the present disclosure provides methods of treating a disease modulated by topoisomerase I in a subject, comprising administering a compound disclosed herein, or a pharmaceutically acceptable salt thereof, to the subject.

[0019] BRIEF DESCRIPTION OF THE DRAWINGS

[0020] FIG. 1 shows the Western blot analysis of TOPI protein levels in HCT15 cells treated with exemplified compounds at 100 nM for 4 hours and 24 hours. Untreated control samples are indicated as ‘NC’. GAPDH was used as a loading control.

[0021] FIG. 2 shows TOPI protein levels in HCT15 cells treated with exemplified compounds at 100 nM for 1, 2, 4, 8, and 24 hours. Untreated control samples are indicated as ‘NC’ . GAPDH was used as a loading control.

[0022] FIG.3 shows the durability of TOPI suppression after exemplified compound washout. HCT15 cells were treated with each compound for 4 h, washed, and incubated in compound- free medium. TOPI protein levels were measured at 0, 1, 2, 4, 8, and 24 h after washout. Untreated control samples are indicated as ‘NC’. GAPDH was used as a loading control.

[0023] DETAILED DESCRIPTION OF THE INVENTION

[0024] In certain aspects, the present disclosure provides indenoisoquinoline-based, non- camptothecin compounds. In certain aspects, the present disclosure provides compounds having a structure represented by Formula I, or a pharmaceutically acceptable salt thereof:

[0025] -2-

[0026] FH13103096.12 Attorney Docket No.: LCH-03225

[0027] I wherein

[0028] R1, R2, R3and R4are each independently H, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, sulfonyl, cycloalkyl, or heterocyclyl; or R2and R3, together with the carbon atoms that separate them, complete a heterocyclyl;

[0029] Y is alkylene, alkenylene, alkynylene, cycloalkylene, heteroalkylene, or heteroalkenylene;

[0030] Z is aryl, heteroaryl, cycloalkyl, heterocyclyl, amido, or amino; and

[0031] A is aryl or heteroaryl.

[0032] In certain embodiments, the present disclosure provides compounds having a structure represented by Formula la, or a pharmaceutically acceptable salt thereof: la wherein

[0033] R1, R2, R3and R4are each independently H, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, cycloalkyl, or heterocyclyl; or R2and R3, together with the carbon atoms that separate them, complete a heterocyclyl;

[0034] Y is alkylene, alkenylene, cycloalkylene, heteroalkylene, or heteroalkenylene;

[0035] Z is aryl, heteroaryl, cycloalkyl, heterocyclyl, amido, or amino; and A is aryl or heteroaryl.

[0036] In certain embodiments, if R2and R3are both H or both methoxy and Z is heteroaryl or heterocyclyl, then Z is substituted with -NRaRbor -alkyl-NRaRb, wherein Raand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

[0037] -3-

[0038] FH13103096.12 Attorney Docket No.: LCH-03225

[0039] In other embodiments, if R2is nitro or halo and Z is heteroaryl or heterocyclyl, then Z is substituted with -NRaRbor -alkyl-NRaRb.

[0040] In certain embodiments, if R2is methoxy and R3is H, then Z is aryl, heteroaryl, cycloalkyl, amido, or amino.

[0041] In other embodiments, if R2or R3is carboxyl and Z is heteroaryl or heterocyclyl, then Z is substituted with -NRaRbor -alkyl-NRaRb.

[0042] In certain embodiments, A is aryl, preferably phenyl. In other embodiments, A is heteroaryl, such as pyridinyl.

[0043] In certain embodiments, the compound has a structure represented by Formula lb, or a pharmaceutically acceptable salt thereof: lb wherein

[0044] X is CH or N; and

[0045] R5and R6are each independently H, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, cycloalkyl, or heterocyclyl; or R5and R6, together with the carbon atoms that separate them, complete a heterocyclyl.

[0046] In certain embodiments, X is CH. In other preferred embodiments, X is N.

[0047] In certain embodiments, R5is alkoxy, preferably methoxy. In other embodiments, R5is halo, preferably fluoro. In yet other embodiments, R5is alkyl (e.g., methyl).

[0048] In certain embodiments, R6is halo, preferably fluoro. In other embodiments, R6is alkoxy, preferably methoxy. In other embodiments, R6is alkyl e.g., methyl). In yet other embodiments, R6is H.

[0049] In certain embodiments, R5and R6, together with the carbon atoms that separate them, complete a heterocycle, such as 1,3-dioxolane.

[0050] In certain embodiments, the compound has a structure represented by Formula Ic or a pharmaceutically acceptable salt thereof:

[0051] -4-

[0052] FH13103096.12 Attorney Docket No.: LCH-03225

[0053] Ic wherein

[0054] R7and R8are each independently H, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, cycloalkyl, or heterocyclyl.

[0055] In other embodiments, the compound has a structure represented by Formula Id or a pharmaceutically acceptable salt thereof:

[0056] Id wherein

[0057] R3is methoxy or cyano; and

[0058] R2is halo, preferably fluoro.

[0059] In certain embodiments, R7and R8are each deuterium. In other embodiments, R7and R8are each independently H, Ci-6 alkyl, Ci-6 alkoxy, C2-6 alkenyl, C2-6 alkynyl, or fluoro. In certain preferred embodiments, R7and R8are each H.

[0060] In certain embodiments, Z is heteroaryl, such as thiazolyl, pyrazolyl, pyridazinyl, imidazolyl, pyridyl, pyrimidyl, pyrazinyl, or triazinyl, each of which is optionally substituted. In other embodiments, Z is heterocyclyl, such as pyrrolidinyl, piperidinyl, piperidinonyl, piperazyl, imidazolidinyl, oxazolidinyl, isothiazolidinyl, hexahydropyridazinyl, morpholinyl, tetrahydropyrrolopyridyl, tetrahydropyrrolopyridinonyl, pyranopyrimidinedionyl, pyrrolopyrimidyl, oxazaspirodecyl, or oxadiazolyl, each of which is optionally substituted. In certain preferred embodiments, Z is pyrazolyl. In other embodiments, Z is

[0061] -5-

[0062] FH13103096.12 Attorney Docket No.: LCH-03225 dihydropyranopyridinedionyl dihydropyranopyrimidinedionyl (e.g., ) tetrahydrooxepinopyridinedionyl (e.g., or tetrahydrotriazinyl

[0063] In certain embodiments, Z is substituted with oxo, amido, alkyl, halo, or hydroxyl, and is optionally further substituted.

[0064] In certain embodiments, Z is heteroaryl or heterocyclyl; Z is substituted with -NRaRbor -alkyl-NRaRb, and is optionally further substituted; and Raand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

[0065] In other embodiments, Z is cycloalkyl, such as cyclobutyl, cyclopentyl, bicyclopentyl, or bicyclohexyl, each of which is optionally substituted. In certain embodiments, Z is cycloalkyl (e.g., bicyclo [l.l.l]pentanyl or bicyclo[2.1.1]hexanyl). In certain embodiments, Z is substituted or unsubstituted alkanecarboxamido, such as morpholinylacetamido, hydroxyacetamido, or aminoacetamido. In certain embodiments, Z is substituted or unsubstituted alkanecarboxamido, such as morpholinylacetamido or hydroxyacetamido.

[0066] In other embodiments, Z is amido. In certain embodiments, “amido”, as used herein, refers to a group

[0067] R9O vy” o , or R10, wherein R9and R10each independently represent a hydrogen or hydrocarbyl group, or R9and R10taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

[0068] In certain embodiments, Z is alkyl-substituted carboxamido. In certain embodiments, Z is substituted with alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, cycloalkyl, and heterocyclyl. In certain embodiments, Z is substituted with hydroxyl, halo such as fluoro, carboxyl, alkoxy such as

[0069] FH13103096.12 Attorney Docket No.: LCH-03225 methoxy, amido such as hydroxymethylamido, sulfonamidyl such as methyl sulfonamidyl, amino, or alkylamino such as methylamino or ethylamino. In certain embodiments, Z is substituted with alkyl (e.g., (fluoro)(hydroxy)methyl or aminoalkyl).

[0070] In certain embodiments, Y is alkylene, such as propylene or butylene, optionally substituted with hydroxyl or dimethylaminopropyl. In certain embodiments, Y is alkylene (e.g., propylene or butylene), optionally substituted with hydroxyl, dimethylaminomethyl, dimethylaminopropyl, or azetidinemethyl. In certain embodiments, Y is -(CH2)t-, wherein t is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0071] In certain preferred embodiments, t is 3. In other preferred embodiments, t is 4.

[0072] In certain embodiments, Y is heteroalkylene. In certain preferred embodiments, Y is *- NHCH2CH2-, *-CH2CH2-NH-, *-CH2CH2O-, *-CH2CH2CH2O-, or *-CH2CH2CH2NH-, wherein * indicates the point of attachment to the nitrogen of formula I. In certain embodiments, Y is alkenylene, such as butenyl-2-ene.

[0073] In other embodiments, Y is cycloalkylene such as methylcyclobutylene.

[0074] In certain embodiments, Y is substituted with alkyl, such as methyl. In certain embodiments, Y is substituted with alkyl e.g., N,N-dimethylaminoethyl or N,N- dimethylaminomethyl). In certain embodiments, Y is substituted with amido (e.g., aminoalkylaminoalkylamido or aminoalkoxyalkylamido). In certain embodiments, Y is substituted with two methyls. In certain embodiments, Y is further substituted with two fluoros. In other embodiments, Y is substituted with hydroxyl. In certain embodiments, Y is disubstituted, preferably on the same carbon, and the substituents combine to form a heterocyclyl, such as an oxetanyl.

[0075] In certain embodiments, the compound has a structure represented by Formula le or a pharmaceutically acceptable salt thereof: wherein

[0076] T is alkyl (e.g., hydroxyalkyl, (hydroxy)(halo)alkyl, cycloalkylalkyl, or heterocyclylalkyl, such as morpholinylalkyl), and

[0077] -7-

[0078] FH13103096.12 Attorney Docket No.: LCH-03225

[0079] R9is H, alkyl (e.g., trifluoroethyl, dimethylaminoethyl), aralkyl (e.g., benzyl or phenylethyl), or cycloalkyl (e.g., cyclopropyl).

[0080] In certain embodiments, the compound has a structure represented by Formula le or a pharmaceutically acceptable salt thereof: wherein

[0081] T is alkyl (e.g., hydroxyalkyl, (hydroxy)(halo)alkyl, cycloalkylalkyl, or heterocyclylalkyl, such as morpholinylalkyl), and

[0082] R9is H, alkyl (e.g., trifluoroethyl), aralkyl (e.g., benzyl or phenylethyl), or cycloalkyl (e.g., cyclopropyl).

[0083] In certain embodiments, T is alkyl (e.g., methyl) substituted with alkoxy (e.g., methylaminoethoxy) alkylamino (e.g., aminoethylmethylamino, dimethylaminoethylamino), or aminocycloalkyl (e.g., aminocyclopropyl). In other embodiments, T is alkyl (e.g., trifluoroethyl, difluoroethyl, propyl, ethyl, methyl) substituted with one, two, or three substituents selected from amino (e.g., dimethylamino), hydroxyl, aryl (e.g., phenyl), and halo (e.g., fluoro).

[0084] In certain embodiments, the compound has a structure represented by Formula If or a pharmaceutically acceptable salt thereof:

[0085] If wherein n ranges from 0 to 5; p ranges from 0 to 3; and

[0086] FH13103096.12 Attorney Docket No.: LCH-03225

[0087] Z is aryl, heteroaryl, cycloalkyl, heterocyclyl, amido, or amino; and

[0088] Raand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, heteroaryl, alkylsulfoxidyl, alkylsulfonyl, or sulfonamido.

[0089] In certain preferred embodiments, n is 0.

[0090] In other preferred embodiments, n is i.

[0091] In certain embodiments, the compound has a structure represented by Formula Ig or a pharmaceutically acceptable salt thereof: wherein

[0092] Xi is C, CH, or N;

[0093] X2, X3, X4 and X5 are each independently CH, CH2, N, NH, O, or S; at least two of Xi, X2, X3, X4 or X5 are C, CH or CH2; each of Xi, X2, X3, X4 and X5 is optionally substituted with W; each W is independently selected from alkyl, hydroxyl, oxo, amido, amino, heteroaryl, and heterocyclyl;

[0094] = is a single bond or a double bond; and m is an integer selected from 1 and 2.

[0095] In certain preferred embodiments, at least one W is NRaRb; and Raand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

[0096] In certain preferred embodiments, m is 1.

[0097] In certain embodiments, Xi, X3 and X5 are N; and X2 and X4 are CH, optionally substituted with W.

[0098] In certain embodiments, the compound has a structure, represented by Formula Ih or a pharmaceutically acceptable salt thereof:

[0099] -9-

[0100] FH13103096.12 Attorney Docket No.: LCH-03225 wherein

[0101] U is alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl. In certain embodiments, the compound has a structure, represented by Formula li or a pharmaceutically acceptable salt thereof: wherein

[0102] U is NRaRb, wherein Raand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

[0103] In other embodiments, the topoisomerase compound has a structure, represented by Formula Ij or a pharmaceutically acceptable salt thereof:

[0104] FH13103096.12 Attorney Docket No.: LCH-03225

[0105] In certain embodiments, two of Xi, X2, X3, X4 and X5 are N or NH, and the remainder are C or CH, optionally substituted with W. In certain preferred embodiments, Xi and X2 are N; and X3, X4, and X5 are CH, optionally substituted with W.

[0106] In certain embodiments, the topoisomerase compound has a structure, represented by Formula Ik or a pharmaceutically acceptable salt thereof:

[0107] Ik wherein

[0108] U is alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

[0109] In certain embodiments, the topoisomerase compound has a structure, represented by Formula II or a pharmaceutically acceptable salt thereof:

[0110] II wherein

[0111] U is hydroxyl or NRaRb, wherein Raand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

[0112] In certain embodiments, U is amido (e.g., aminoalkylamido, aminoalkylaminoalkylamido, carboxyalkylamido, amidoalkylamido, cycloalkylamido, such as aminocycloalkylamido, or hydrazineamido).

[0113] In certain preferred embodiments, the topoisomerase compound has a structure, represented by Formula Im or a pharmaceutically acceptable salt thereof:

[0114] -11-

[0115] FH13103096.12 Attorney Docket No.: LCH-03225

[0116] In certain embodiments, Rais H. In other embodiments, Rais alkyl, such as methyl or trifluoroethyl. In certain embodiments, Rais alkyl (e.g., ethyl).

[0117] In certain embodiments, Rbis heterocyclyl, such as piperidinyl or pyrrolidinyl. In other embodiments, Rbis alkyl, such as methyl, ethyl, propyl, butyl, or pentyl. In certain embodiments, Rbis alkyl, such as cyclobutylmethyl cyclopentylmethyl, or cyclobutylethyl. In certain embodiments, wherein Rbis alkyl sulfonyl, such as methylsulfonyl. In other embodiments, Rbis heteroaryl, such as pyridinyl. In yet other embodiments, Rbis amido further substituted with amino, such as alkylamino (e.g., Rbis alkylN(H)N(H)C(O)- or methylN(H)N(H)C(O)-). In yet other embodiments, Rbis acyl. In yet other embodiments, Rbis polyethylene glycolyl (PEG). In yet other embodiments, Rbis H.

[0118] In certain embodiments, Rbis optionally substituted with one or more substituents selected from alkyl, hydroxyl, oxo, halo, alkoxy, and amino. In certain embodiments, Rbis optionally substituted with one, two, or three substituents selected from alkyl (e.g., hydroxymethyl), hydroxyl, oxo, halo, alkoxy, amino, alkylamino (e.g., dimethylamino, methylamino), cycloalkyl (e.g., cyclopropyl), amido (e.g., methylamido), and heterocycloalkyl (e.g., azetidinyl or oxetanyl). In certain embodiments, Rbis optionally substituted with one, two, or three selected from alkyl, hydroxyl, oxo, halo, alkoxy, and amino. In certain embodiments, the heterocycloalkyl on Rbis substituted with halo, hydroxyl, alkyl (e.g., trifluoromethyl), carboxyl, amino, or amido (e.g., methylamido). In certain embodiments, Rbis substituted with alkyl, such as methyl or trifluoromethyl. In certain embodiments, Rbis substituted with alkyl (e.g., cyclopropylmethyl) and amino. In certain embodiments, Rbis substituted with hydroxyl. In certain embodiments, Rbis substituted with oxo. In certain embodiments, Rbis substituted with halo, such as fluoro. In certain embodiments, Rbis substituted with alkoxy, such as methoxy. In certain embodiments, Rbis substituted with amino. In certain embodiments, Rbis substituted with hydroxyl and halo, such as fluoro. In certain embodiments, Rbis substituted with hydroxyl and alkyl, such as methyl, trifluoromethyl, cyclobutylmethyl, or cyclopropylmethyl. In certain embodiments, Rbis substituted with oxo and alkoxy, such as methoxy. In certain embodiments, Rbis substituted

[0119] -12-

[0120] FH13103096.12 Attorney Docket No.: LCH-03225 with oxo and hydroxyl. In certain embodiments, Rbis substituted with hydroxyl, cycloalkyl, such as cyclopentyl, and halo, such as fluoro. In certain embodiments, Rbis substituted with hydroxyl and cycloalkyl, such as cyclopropyl. In certain embodiments, Rbis substituted with amino and halo, such as difluoro. In certain embodiments, Rbis substituted with amino and cycloalkyl, such as cyclobutyl, cyclopentyl, or cyclopropyl. In certain embodiments, Rbis substituted with alkyl, such as trifluoromethyl or dimethylaminopropyl, and amino. In certain embodiments, Rbis substituted with aminoalkoxy. In certain embodiments, the aminoalkoxy on Rbis an amino group substituted with an alkoxy group. In certain embodiments, Rbis substituted with amino, hydroxyl, and alkyl (e.g., methyl).

[0121] In certain embodiments, R2and R3are each independently alkoxy, such as methoxy, difluoromethoxy, ethoxy, difluoroethoxy, trifluoroethoxy, isopropoxy, or carboxymethoxy, halo, such as fluoro or chloro, or cyano. In other embodiments, R2is halo, preferably fluoro. In certain embodiments, R3is alkoxy, preferably methoxy, difluoromethoxy, or difluoroethoxy. In certain embodiments, R2is halo, preferably fluoro, and R3is methoxy. In certain embodiments, R2is halo, preferably fluoro, and R3is cyano. In certain embodiments, R2and R3are each independently H or alkoxy, such as methoxy. In certain embodiments, R2and R3are each independently H or halo, such as fluoro. In certain embodiments, R2and R3are each independently alkoxy, such as methoxy or difluoromethoxy. In certain embodiments, R2and R3are each independently halo, such as fluoro or chloro, or cyano. In certain embodiments, R2and R3are each independently alkoxy, such as methoxy or difluoromethoxy, or cyano. In certain embodiments, R2and R3are each independently hydroxyl or cyano. In certain embodiments, R2and R3are each independently alkoxy, such as methoxy, or amino, such as - NH2 or difluoroethylamino. In certain embodiments, R2and R3are each independently alkoxy, such as methoxy, or nitro. In certain embodiments, R2and R3are each independently alkoxy, such as methoxy, or amido, such as acetamido or difluoroacetamido. In certain embodiments, R2and R3are each independently alkoxy, such as methoxy, or alkyl, such as trifluoromethyl. In certain embodiments, R2and R3are each independently halo, such as fluoro, or alkyl, such as trifluoromethyl. In certain embodiments, R2and R3are each independently halo, such as fluoro, or hydroxyl. In certain embodiments, R2and R3are each independently halo, such as fluoro, or amido. In certain embodiments, R2and R3combine to form a heterocycle, such as 1,3 -dioxolane.

[0122] In certain embodiments, the compound is selected from:

[0123] -13-

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[0133] -22-

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[0135] -23-

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[0138] -25-

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[0141] -27-

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[0146] FH13103096.12 Attorney Docket No.: LCH-03225 or a pharmaceutically acceptable salt thereof.

[0147] In other aspects, the present disclosure provides pharmaceutical compositions comprising a compound disclosed herein and a pharmaceutically acceptable excipient.

[0148] In yet other aspects, the present disclosure provides methods of treating a proliferative disorder in a subject, comprising administering a compound disclosed herein, or a pharmaceutically acceptable salt thereof, to the subject. In certain embodiments, the proliferative disease is cancer. In certain embodiments, the proliferative disease is selected from breast cancer, lung cancer, bladder cancer, brain cancer, head and neck cancer, liver cancer, biliary tract cancer, cervical cancer, endometrial cancer, esophageal cancer, carcinomas, acute and chronic lymphoid leukemias, acute and chronic myelogenous leukemias, chronic myelomonocytic leukemias, colorectal cancer, gastric cancer, gastrointestinal stromal cancer, glioma, lymphomas, melanomas, multiple myeloma, myeloproliferative diseases, neuroendocrine diseases, pancreatic cancer, ovarian cancer, prostate cancer, renal cell cancer, sarcomas, thyroid cancer, mast cell leukemia, germ cell tumors, testicular cancer, small-cell lung carcinoma, neuroblastoma, osteosarcoma, and retinoblastoma. In certain embodiments, the proliferative disease is selected from breast cancer, lung cancer, bladder cancer, brain cancer, head and neck cancer, liver cancer, biliary tract cancer, carcinomas, acute and chronic lymphoid leukemias, acute and chronic myelogenous leukemias, chronic myelomonocytic -32-

[0149] FH13103096.12 Attorney Docket No.: LCH-03225 leukemias, colorectal cancer, gastric cancer, gastrointestinal stromal cancer, glioma, lymphomas, melanomas, multiple myeloma, myeloproliferative diseases, neuroendocrine diseases, pancreatic cancer, ovarian cancer, prostate cancer, renal cell cancer, sarcomas, thyroid cancer, mast cell leukemia, germ cell tumors, small-cell lung carcinoma, neuroblastoma, and osteosarcoma.

[0150] In yet other aspects, the present disclosure provides methods of inhibiting topoisomerase I in a cell in vitro, comprising contacting the cell with a compound disclosed herein, or a pharmaceutically acceptable salt thereof.

[0151] Pharmaceutical Compositions

[0152] The compositions and methods of the present invention may be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or the compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are for human administration, particularly for invasive routes of administration (i.e., routes, such as injection or implantation, that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipients can be chosen, for example, to effect delayed release of an agent or to selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form such as tablet, capsule (including sprinkle capsule and gelatin capsule), granule, lyophile for reconstitution, powder, solution, syrup, suppository, injection or the like. The composition can also be present in a transdermal delivery system, e.g., a skin patch. The composition can also be present in a solution suitable for topical administration, such as a lotion, cream, or ointment.

[0153] A pharmaceutically acceptable carrier can contain physiologically acceptable agents that act, for example, to stabilize, increase solubility or to increase the absorption of a compound such as a compound of the invention. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a

[0154] -33-

[0155] FH13103096.12 Attorney Docket No.: LCH-03225 physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) also can be a liposome or other polymer matrix, which can have incorporated therein, for example, a compound of the invention. Liposomes, for example, which comprise phospholipids or other lipids, are non-toxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.

[0156] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0157] The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0158] A pharmaceutical composition (preparation) can be administered to a subject by any of a number of routes of administration including, for example, orally (for example, drenches as in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (for example as a patch applied to the skin); and topically (for example, as a

[0159] -34-

[0160] FH13103096.12 Attorney Docket No.: LCH-03225 cream, ointment or spray applied to the skin). The compound may also be formulated for inhalation. In certain embodiments, a compound may be simply dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable for same can be found in, for example, U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, as well as in patents cited therein.

[0161] The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about ninety-nine percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

[0162] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0163] Formulations of the invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), lyophile, powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. Compositions or compounds may also be administered as a bolus, electuary or paste.

[0164] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as

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[0166] FH13103096.12 Attorney Docket No.: LCH-03225 starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as, modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

[0167] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surfaceactive or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0168] The tablets, and other solid dosage forms of the pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances

[0169] -36-

[0170] FH13103096.12 Attorney Docket No.: LCH-03225 and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

[0171] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

[0172] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0173] Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0174] Dosage forms for the topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required.

[0175] The ointments, pastes, creams and gels may contain, in addition to an active compound, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0176] Powders and sprays can contain, in addition to an active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0177] Transdermal patches have the added advantage of providing controlled delivery of a compound of the present invention to the body. Such dosage forms can be made by dissolving

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[0179] FH13103096.12 Attorney Docket No.: LCH-03225 or dispersing the active compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

[0180] The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

[0181] Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0182] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

[0183] In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of

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[0185] FH13103096.12 Attorney Docket No.: ECH-03225 dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0186] Injectable depot forms are made by forming microencapsulated matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissue.

[0187] For use in the methods of this invention, active compounds can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.

[0188] Methods of introduction may also be provided by rechargeable or biodegradable devices. Various slow-release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form an implant for the sustained release of a compound at a particular target site.

[0189] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0190] The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound(s) being employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0191] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the pharmaceutical composition or compound at levels lower than that required in order to achieve the desired therapeutic effect

[0192] -39-

[0193] FH13103096.12 Attorney Docket No.: LCH-03225 and gradually increase the dosage until the desired effect is achieved. By “therapeutically effective amount” is meant the concentration of a compound that is sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the weight, sex, age, and medical history of the subject. Other factors which influence the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound of the invention. A larger total dose can be delivered by multiple administrations of the agent. Methods to determine efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13 ed., 1814-1882, herein incorporated by reference).

[0194] In general, a suitable daily dose of an active compound used in the compositions and methods of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0195] If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In certain embodiments of the present invention, the active compound may be administered two or three times daily. In preferred embodiments, the active compound will be administered once daily.

[0196] The patient receiving this treatment is any animal in need, including primates, in particular humans; and other mammals such as equines, cattle, swine, sheep, cats, and dogs; poultry; and pets in general.

[0197] In certain embodiments, compounds of the invention may be used alone or conjointly administered with another type of therapeutic agent.

[0198] The present disclosure includes the use of pharmaceutically acceptable salts of compounds of the invention in the compositions and methods of the present invention. In certain embodiments, contemplated salts of the invention include, but are not limited to, alkyl, dialkyl, trialkyl or tetra-alkyl ammonium salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, L-arginine, benethamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, IH-imidazole, lithium, L- lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, l-(2-

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[0200] FH13103096.12 Attorney Docket No.: LCH-03225 hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, l-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2- hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, 1-ascorbic acid, 1-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1 ,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, 1-malic acid, malonic acid, mandelic acid, methanesulfonic acid , naphthalene- 1,5 -disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, 1-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, 1-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid acid salts.

[0201] The pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent.

[0202] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0203] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

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[0205] FH13103096.12 Attorney Docket No.: LCH-03225

[0206] Definitions

[0207] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art. Generally, nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics and protein and nucleic acid chemistry, described herein, are those well-known and commonly used in the art.

[0208] The methods and techniques of the present disclosure are generally performed, unless otherwise indicated, according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout this specification. See, e.g. “Principles of Neural Science”, McGraw-Hill Medical, New York, N.Y. (2000); Motulsky, “Intuitive Biostatistics”, Oxford University Press, Inc. (1995); Lodish et al., “Molecular Cell Biology, 4th ed.”, W. H. Freeman & Co., New York (2000); Griffiths et al., “Introduction to Genetic Analysis, 7th ed.”, W. H. Freeman & Co., N.Y. (1999); and Gilbert et al., “Developmental Biology, 6th ed.”, Sinauer Associates, Inc., Sunderland, MA (2000).

[0209] Chemistry terms used herein, unless otherwise defined herein, are used according to conventional usage in the art, as exemplified by “The McGraw-Hill Dictionary of Chemical Terms”, Parker S., Ed., McGraw-Hill, San Francisco, C.A. (1985).

[0210] All of the above, and any other publications, patents and published patent applications referred to in this application are specifically incorporated by reference herein. In case of conflict, the present specification, including its specific definitions, will control.

[0211] The term “agent” is used herein to denote a chemical compound (such as an organic or inorganic compound, a mixture of chemical compounds), a biological macromolecule (such as a nucleic acid, an antibody, including parts thereof as well as humanized, chimeric and human antibodies and monoclonal antibodies, a protein or portion thereof, e.g., a peptide, a lipid, a carbohydrate), or an extract made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues. Agents include, for example, agents whose structure is known, and those whose structure is not known.

[0212] A “patient,” “subject,” or “individual” are used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (including bovines, porcines, etc.), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats).

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[0214] FH13103096.12 Attorney Docket No.: LCH-03225

[0215] “Treating” a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0216] The term “preventing” is art-recognized, and when used in relation to a condition, such as a local recurrence (e.g., pain), a disease such as cancer, a syndrome complex such as heart failure or any other medical condition, is well understood in the art, and includes administration of a composition which reduces the frequency of, or delays the onset of, symptoms of a medical condition in a subject relative to a subject which does not receive the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable cancerous growths in a treated population versus an untreated control population, e.g., by a statistically and / or clinically significant amount.

[0217] “Administering” or “administration of’ a substance, a compound or an agent to a subject can be carried out using one of a variety of methods known to those skilled in the art. For example, a compound or an agent can be administered, intravenously, arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (by absorption, e.g., through a skin duct). A compound or agent can also appropriately be introduced by rechargeable or biodegradable polymeric devices or other devices, e.g., patches and pumps, or formulations, which provide for the extended, slow or controlled release of the compound or agent. Administering can also be performed, for example, once, a plurality of times, and / or over one or more extended periods.

[0218] Appropriate methods of administering a substance, a compound or an agent to a subject will also depend, for example, on the age and / or the physical condition of the subject and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability and toxicity). In some embodiments, a compound or an agent is administered orally, e.g., to a subject by ingestion. In some embodiments, the orally

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[0220] FH13103096.12 Attorney Docket No.: LCH-03225 administered compound or agent is in an extended release or slow release formulation, or administered using a device for such slow or extended release.

[0221] As used herein, the phrase “conjoint administration” refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered therapeutic agent is still effective in the body (e.g., the two agents are simultaneously effective in the patient, which may include synergistic effects of the two agents). For example, the different therapeutic compounds can be administered either in the same formulation or in separate formulations, either concomitantly or sequentially. Thus, an individual who receives such treatment can benefit from a combined effect of different therapeutic agents.

[0222] A “therapeutically effective amount” or a “therapeutically effective dose” of a drug or agent is an amount of a drug or an agent that, when administered to a subject will have the intended therapeutic effect. The full therapeutic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount needed for a subject will depend upon, for example, the subject’s size, health and age, and the nature and extent of the condition being treated, such as cancer or MDS. The skilled worker can readily determine the effective amount for a given situation by routine experimentation.

[0223] As used herein, the terms “optional” or “optionally” mean that the subsequently described event or circumstance may occur or may not occur, and that the description includes instances where the event or circumstance occurs as well as instances in which it does not. For example, “optionally substituted alkyl” refers to the alkyl may be substituted as well as where the alkyl is not substituted.

[0224] It is understood that substituents and substitution patterns on the compounds of the present invention can be selected by one of ordinary skilled person in the art to result chemically stable compounds which can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results.

[0225] As used herein, the term “optionally substituted” refers to the replacement of one to six hydrogen radicals in a given structure with the radical of a specified substituent including, but not limited to: hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl,

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[0227] FH13103096.12 Attorney Docket No.: LCH-03225 cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O- alkyl, -OP(O)(O-alkyl)2 or -CH2-OP(O)(O-alkyl)2. Preferably, “optionally substituted” refers to the replacement of one to four hydrogen radicals in a given structure with the substituents mentioned above. More preferably, one to three hydrogen radicals are replaced by the substituents as mentioned above. It is understood that the substituent can be further substituted.

[0228] As used herein, the term “alkyl” refers to saturated aliphatic groups, including but not limited to C1-C10 straight-chain alkyl groups or C1-C10 branched-chain alkyl groups. Preferably, the “alkyl” group refers to Ci-Ce straight-chain alkyl groups or Ci-Ce branched- chain alkyl groups. Most preferably, the “alkyl” group refers to C1-C4 straight-chain alkyl groups or C1-C4 branched-chain alkyl groups. Examples of “alkyl” include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, neo-pentyl, 1 -hexyl, 2-hexyl, 3-hexyl, 1 -heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1 -octyl, 2-octyl, 3-octyl or 4-octyl and the like. The “alkyl” group may be optionally substituted.

[0229] The term “acyl” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0230] The term “acylamino” is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbylC(O)NH-.

[0231] The term “acyloxy” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0232] The term “alkoxy” refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like.

[0233] The term “alkoxyalkyl” refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

[0234] The term “alkyl” refers to saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In preferred embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., Ci- 30 for straight chains, C3-30 for branched chains), and more preferably 20 or fewer.

[0235] Moreover, the term “alkyl” as used throughout the specification, examples, and claims is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2- trifluoroethyl, etc.

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[0237] FH13103096.12 Attorney Docket No.: LCH-03225

[0238] The term “Cx.y” or “Cx-Cy”, when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups that contain from x to y carbons in the chain. Coalkyl indicates a hydrogen where the group is in a terminal position, a bond if internal. A Ci-ealkyl group, for example, contains from one to six carbon atoms in the chain.

[0239] The term “alkylamino”, as used herein, refers to an amino group substituted with at least one alkyl group.

[0240] The term “alkylthio”, as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

[0241] The term “amido”, as used herein, refers to a group wherein R9and R10each independently represent a hydrogen or hydrocarbyl group, or R9and R10taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

[0242] The terms “amine” and “amino” are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by wherein R9, R10, and R10’ each independently represent a hydrogen or a hydrocarbyl group, or R9and R10taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

[0243] The term “aminoalkyl”, as used herein, refers to an alkyl group substituted with an amino group.

[0244] The term “aralkyl”, as used herein, refers to an alkyl group substituted with an aryl group.

[0245] The term “aryl” as used herein includes substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon. Preferably the ring is a 5- to 7- membered ring, more preferably a 6-membered ring. The term “aryl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can

[0246] -46-

[0247] FH13103096.12 Attorney Docket No.: LCH-03225 be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0248] The term “carbamate” is art-recognized and refers to a group

[0249] O O

[0250] A JL pio A u pio

[0251] 'Sr' 'HorN O'R

[0252] R9R9wherein R9and R10independently represent hydrogen or a hydrocarbyl group.

[0253] The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

[0254] The term “carbocycle” includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated and aromatic rings. Carbocycle includes bicyclic molecules in which one, two or three or more atoms are shared between the two rings. The term “fused carbocycle” refers to a bicyclic carbocycle in which each of the rings shares two adjacent atoms with the other ring. Each ring of a fused carbocycle may be selected from saturated, unsaturated and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, is included in the definition of carbocyclic. Exemplary “carbocycles” include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5 -cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct- 3-ene, naphthalene and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-lH- indene and bicyclo[4.1.0]hept-3-ene. “Carbocycles” may be substituted at any one or more positions capable of bearing a hydrogen atom.

[0255] The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

[0256] The term “carbonate” is art-recognized and refers to a group -OCO2-.

[0257] The term “carboxy”, as used herein, refers to a group represented by the formula -CO2H.

[0258] The term “cycloalkyl” includes substituted or unsubstituted non-aromatic single ring structures, preferably 4- to 8-membered rings, more preferably 4- to 6-membered rings. The term “cycloalkyl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is cycloalkyl and the substituent (e.g., R100) is attached to the cycloalkyl ring, e.g., the other -47-

[0259] FH13103096.12 Attorney Docket No.: LCH-03225 cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, benzodioxane, tetrahydroquinoline, and the like.

[0260] The term “ester”, as used herein, refers to a group -C(O)OR9wherein R9represents a hydrocarbyl group.

[0261] The term “ether”, as used herein, refers to a hydrocarbyl group linked through an oxygen to another hydrocarbyl group. Accordingly, an ether substituent of a hydrocarbyl group may be hydrocarbyl-O-. Ethers may be either symmetrical or unsymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include “alkoxyalkyl” groups, which may be represented by the general formula alkyl-O-alkyl.

[0262] The terms “halo” and “halogen” as used herein means halogen and includes chloro, fluoro, bromo, and iodo.

[0263] The terms “hetaralkyl” and “heteroaralkyl”, as used herein, refers to an alkyl group substituted with a hetaryl group.

[0264] The terms “heteroaryl” and “hetaryl” include substituted or unsubstituted aromatic single ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heteroaryl” and “hetaryl” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.

[0265] The term “heteroatom” as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0266] The term “heterocyclylalkyl”, as used herein, refers to an alkyl group substituted with a heterocycle group.

[0267] The terms “heterocyclyl”, “heterocycle”, and “heterocyclic” refer to substituted or unsubstituted non-aromatic ring structures, preferably 3- to 10-membered rings, more preferably 3- to 7-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heterocyclyl” and “heterocyclic” also include polycyclic ring systems having two or more

[0268] -48-

[0269] FH13103096.12 Attorney Docket No.: LCH-03225 cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

[0270] The term “hydrocarbyl”, as used herein, refers to a group that is bonded through a carbon atom that does not have a =0 or =S substituent, and typically has at least one carbonhydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Thus, groups like methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered to be hydrocarbyl for the purposes of this application, but substituents such as acetyl (which has a =0 substituent on the linking carbon) and ethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbyl groups include, but are not limited to aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

[0271] The term “hydroxyalkyl”, as used herein, refers to an alkyl group substituted with a hydroxy group.

[0272] The term “lower” when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups where there are ten or fewer atoms in the substituent, preferably six or fewer. A “lower alkyl”, for example, refers to an alkyl group that contains ten or fewer carbon atoms, preferably six or fewer. In certain embodiments, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are respectively lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, such as in the recitations hydroxyalkyl and aralkyl (in which case, for example, the atoms within the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

[0273] The terms “polycyclyl”, “polycycle”, and “polycyclic” refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and / or heterocyclyls) in which two or more atoms are common to two adjoining rings, e.g., the rings are “fused rings”. Each of the rings of the polycycle can be substituted or unsubstituted. In certain embodiments, each ring of the polycycle contains from 3 to 10 atoms in the ring, preferably from 5 to 7.

[0274] The term “sulfate” is art-recognized and refers to the group -OSO3H, or a pharmaceutically acceptable salt thereof.

[0275] The term “sulfonamide” is art-recognized and refers to the group represented by the general formulae

[0276] -49-

[0277] FH13103096.12 Attorney Docket No.: LCH-03225 wherein R9and R10independently represent hydrogen or hydrocarbyl.

[0278] The term “sulfoxide” is art-recognized and refers to the group-S(O)-.

[0279] The term “sulfonate” is art-recognized and refers to the group SO3H, or a pharmaceutically acceptable salt thereof.

[0280] The term “sulfone” is art-recognized and refers to the group -S(O)2-.

[0281] The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxy carbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamide, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.

[0282] The term “thioalkyl”, as used herein, refers to an alkyl group substituted with a thiol group.

[0283] The term “thioester”, as used herein, refers to a group -C(O)SR9or -SC(O)R9wherein R9represents a hydrocarbyl.

[0284] The term “thioether”, as used herein, is equivalent to an ether, wherein the oxygen is replaced with a sulfur.

[0285] -50-

[0286] FH13103096.12 Attorney Docket No.: ECH-03225

[0287] The term “urea” is art-recognized and may be represented by the general formula wherein R9and R10independently represent hydrogen or a hydrocarbyl.

[0288] The term “modulate” as used herein includes the inhibition or suppression of a function or activity (such as cell proliferation) as well as the enhancement of a function or activity.

[0289] The phrase “pharmaceutically acceptable” is art-recognized. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0290] “Pharmaceutically acceptable salt” or “salt” is used herein to refer to an acid addition salt or a basic addition salt which is suitable for or compatible with the treatment of patients.

[0291] The term “pharmaceutically acceptable acid addition salt” as used herein means any non-toxic organic or inorganic salt of any base compounds represented by Formula I. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric and phosphoric acids, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, benzoic, phenylacetic, cinnamic and salicylic acids, as well as sulfonic acids such as p-toluene sulfonic and methanesulfonic acids. Either the mono or di-acid salts can be formed, and such salts may exist in either a hydrated, solvated or substantially anhydrous form. In general, the acid addition salts of compounds of Formula I are more soluble in water and various hydrophilic organic solvents, and generally demonstrate higher melting points in comparison to their free base forms. The selection of the appropriate salt will be known to one skilled in the art. Other non- pharmaceutically acceptable salts, e.g., oxalates, may be used, for example, in the isolation of compounds of Formula I for laboratory use, or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0292] The term “pharmaceutically acceptable basic addition salt” as used herein means any non-toxic organic or inorganic base addition salt of any acid compounds represented by Formula I or any of their intermediates. Illustrative inorganic bases which form suitable salts include lithium, sodium, potassium, calcium, magnesium, or barium hydroxide. Illustrative -51-

[0293] FH13103096.12 Attorney Docket No.: LCH-03225 organic bases which form suitable salts include aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine and picoline or ammonia. The selection of the appropriate salt will be known to a person skilled in the art.

[0294] Many of the compounds useful in the methods and compositions of this disclosure have at least one stereogenic center in their structure. This stereogenic center may be present in a R or a S configuration, said R and S notation is used in correspondence with the rules described in Pure Appl. Chem. (1976), 45, 11-30. The disclosure contemplates all stereoisomeric forms such as enantiomeric and diastereoisomeric forms of the compounds, salts, prodrugs or mixtures thereof (including all possible mixtures of stereoisomers). See, e.g., WO 01 / 062726.

[0295] Furthermore, certain compounds which contain alkenyl groups may exist as Z (zusammen) or E (entgegen) isomers. In each instance, the disclosure includes both mixture and separate individual isomers.

[0296] “Prodrug” or “pharmaceutically acceptable prodrug” refers to a compound that is metabolized, for example hydrolyzed or oxidized, in the host after administration to form the compound of the present disclosure (e.g., compounds of formula I). Typical examples of prodrugs include compounds that have biologically labile or cleavable (protecting) groups on a functional moiety of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to produce the active compound. Examples of prodrugs using ester or phosphoramidate as biologically labile or cleavable (protecting) groups are disclosed in U.S. Patents 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. The prodrugs of this disclosure are metabolized to produce a compound of Formula I. The present disclosure includes within its scope, prodrugs of the compounds described herein. Conventional procedures for the selection and preparation of suitable prodrugs are described, for example, in “Design of Prodrugs” Ed. H. Bundgaard, Elsevier, 1985.

[0297] The phrase “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filter, diluent, excipient, solvent or encapsulating material useful for formulating a drug for medicinal or therapeutic use.

[0298] The term “Log of solubility”, “LogS” or “logS” as used herein is used in the art to quantify the aqueous solubility of a compound. The aqueous solubility of a compound significantly affects its absorption and distribution characteristics. A low solubility often goes

[0299] -52-

[0300] FH13103096.12 Attorney Docket No.: LCH-03225 along with a poor absorption. LogS value is a unit stripped logarithm (base 10) of the solubility measured in mol / liter.

[0301] EXAMPLES

[0302] The invention now being generally described, it will be more readily understood by reference to the following examples which are included merely for purposes of illustration of certain aspects and embodiments of the present invention and are not intended to limit the invention.

[0303] Preparation Example 1: Synthesis of intermediate A series

[0304] [Preparative example A- 1] 5,6-dihydroxyisobenzofuran-l(3H)-one (Int. A-l)

[0305] (Step 1) 5 ,6-dimethoxyisobenzofuran- 1 (3H)-one

[0306] A mixture of 3,4-dimethoxybenzoic acid (20.0 g, 109 mmol) and 37 wt% formaldehyde aqueous solution (20.2 mL) in cone. HC1 (38 mL) was stirred at 90 °C for 12 h under nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure on completion. The residue was diluted with distilled water (100 mL), then extracted twice with ethyl acetate (EA) (100 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was triturated with n-hexane, filtered, washed with n-hexane, then dried in vacuo to obtain the tide compound as a lightyellow solid (11.6 g, 55%).

[0307] MS m / z: 195 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.32 (s, 1 H), 6.90 (s, 1 H), 5.23 (s, 2 H), 3.96 (s, 3H), 3.95 (s, 3H).

[0308] (Step 2) 5 ,6-dihydroxyisobenzofuran- 1 (3H)-one

[0309] The compound prepared in (step 1) above (3.50 g, 18.1 mmol) was dissolved in DCM (30 mL), then 1 M BBn solution in DCM (48 mL, 48.1 mmol) was added thereto slowly at - 78 °C. The reaction mixture was stirred at RT for 3 h under nitrogen atmosphere. After being cooled to 0 °C, distilled water was added to the reaction mixture. The resulting solid was filtered, washed with distilled water, then dried in vacuo to obtain the title compound as alight- yellow solid (2.9 g, 97%), which was used for the next step without further purification.

[0310] MS m / z: 167 [M+l]+. ’ H NMR (400 MHz, DMSO-&) 57.06 (s, 1H), 6.93 (s, 1H), 5.17

[0311] (s, 2H).

[0312] FH13103096.12 Attorney Docket No.: LCH-03225

[0313] [Preparative example A-2] [l,3]dioxolo[4,5-f]isobenzofuran-5(7H)-one (Int. A-2)

[0314] I nt. A-2

[0315] To a solution of Int. A-l (787 mg, 4.73 mmol) in DMF (15 mL) were added K2CO3 (1.31 g, 9.47 mmol) and dibromomethane (0.4 mL, 5.68 mmol) sequentially at 0 °C, and the mixture was stirred at 60 °C for 14 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as a light- yellow solid (570 mg, 68%).

[0316] MS m / z: 179 [M+l]+. ’ H NMR (400 MHz, CDCI3) 5 7.23 (s, 1H), 6.84 (s, 1H), 6.12(s, 2H), 5.19 (s, 2H).

[0317] [Preparative example A-31 [l,3]dioxolo[4,5-f]isobenzofuran-5(7H)-one-2,2-d2 (Int. A-3)

[0318] The title compound (200 mg, 61%) was synthesized in the same manner as described in preparative example A-2 using Int.A-1 (300 mg, 1.81 mg) and dibromomethan-tfc instead of dibromomethane.

[0319] ‘ H NMR (400 MHz, CDCI3) 5 7.26 (s, 1H), 7.23 (s, 1H), 5.19 (d, J = 0.9 Hz, 2H).

[0320] [Preparative example A-4] 2,2-difluoro-[l,3]dioxolo[4,5-f]isobenzofuran-5(7H)-one (Int. A-

[0321] 4)

[0322] A mixture of 2,2-difluorobenzo[d][l,3]dioxole-5-carboxylic acid (300 mg, 1.48 mmol), Pd(OAc)2 (31.8 mg, 0.15 mmol), K2HPO4 (775 mg, 4.45 mmol), and dibromomethane (258 mg, 1.48 mmol) was stirred at 140 °C for 12 h under nitrogen atmosphere. After being cooled to RT, the reaction mixture was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 4) to obtain the title compound as a light-yellow solid

[0323] -54-

[0324] FH13103096.12 Attorney Docket No.: LCH-03225

[0325] (40.0 mg, 12%).

[0326] MS m / z: 215 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.56 (d, J = 3.8 Hz, 1H), 7.20 (s, 1H), 5.31 (d, 7 = 3.5 Hz, 2H).

[0327] [Preparative example A-51 6-fluoro-5-methoxyisobenzofuran-l(3H)-one (Int. A-5)

[0328] (Step 1) (4-fluoro-3-methoxyphenyl)methanol

[0329] To a solution of 4-fluoro-5-methoxybenzoic acid (1.03 g, 6.05 mmol) in THF (30 mL) was added 1 M BH3 THF complex solution in THF (30.3 mL, 30.3 mmol) at 0 °C, then the mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 5) to obtain the title compound as a colorless oil (921 mg, 97%).

[0330] ’ H NMR (400 MHz, CDCh) 5 7.09 - 6.98 (m, 2H), 6.89 - 6.83 (m, 1H), 4.65 (d, J = 5.4 Hz, 2H), 3.90 (s, 3H).

[0331] (Step 2) (2-bromo-4-fluoro-5 -methoxyphenyl)methanol

[0332] The compound prepared in (step 1) above (1.00 g, 6.40 mmol) was dissolved in (acetonitrile) ACN (15 mL), and NBS (1.03 g, 5.76 mmol) was added thereto portionwise with stirring at 0 °C. The reaction mixture was stirred at RT for 12 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n- hexane = 1 : 2) to obtain the tide compound as a white solid (1.10 g, 73%).

[0333] MS m / z: 236 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.28 (dd, 7 = 10.1, 2.9 Hz, 1H), 7.15 (dd, J = 8.9, 2.9 Hz, 1H), 4.70 (d, J = 2.6 Hz, 2H), 3.91 (d, J = 2.9 Hz, 3H).

[0334] (Step 3) 6-fluoro-5-methoxyisobenzofuran-l(3H)-one (Int. A-5)

[0335] A mixture of the compound prepared in (step 2) above (1.06 g, 4.52 mmol), PdCh(dppf) (662 mg, 0.90 mmol), and DIPEA (1.6 mL, 9.05 mmol) in DMF (14 mL) was stirred at 80 °C for 15 h under carbon monoxide atmosphere (1 atm). On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (100 mL), -55-

[0336] FH13103096.12 Attorney Docket No.: LCH-03225 then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 4 to 1 : 1) to obtain the title compound as a brown solid (502 mg, 65%).

[0337] MS m / z: 183 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.58 (d, J = 9.3 Hz, 1H), 7.00 (d, 7 = 6.9 Hz, 1H), 5.25 (s, 2H), 3.99 (s, 3H).

[0338] [Preparative example A-61 4-fluoro-5-methoxyisobenzofuran-l(3H)-one (Int. A-6)

[0339] The title compound was synthesized in the same manner as described in preparative example A-5 using 2-fluoro-3-methoxybenzoic acid instead of 4-fluoro-5 -methoxybenzoic acid.

[0340] MS m / z: 183 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.70 (d, J = 8.4 Hz, 1H), 7.18 - 7.11 (m, 1H), 5.33 (s, 2H), 4.00 (s, 3H).

[0341] [Preparative example A-7] 6-fluoro-5-methylisobenzofuran-l(3H)-one (Int. A-7)

[0342] The title compound was synthesized in the same manner as described in preparative example A-5 using 2-bromo-4-fluoro-5 -methyl-benzoic acid.

[0343] MS m / z: 167 [M+l]+. ’ H NMR (400 MHz, CDCh) 5= 7.53 (d, 7 = 8.0 Hz, 1H), 7.32 (d, 7 = 6.1 Hz, 1H), 5.26 (s, 2H), 2.42 (d, 7 = 2.1 Hz, 3H).

[0344] [Preparative example A-81 6-chloro-5-methoxyisobenzofuran-l(3H)-one (Int. A-8)

[0345] (Step 1) (4-chloro-3-methoxyphenyl)methanol

[0346] To a solution of 4-chloro-3-methoxybenzoic acid (5.70 g, 30.5 mmol) in THF (100 mL) was added 1 M LiAlH4 solution in THF (36.7 mL, 36.7 mmol) at 0 °C, and the mixture was stirred at the same temperature for 3 h under nitrogen atmosphere. After quenching by adding distilled water (100 mL), 15% NaOH aqueous solution (100 mL) was added to the reaction mixture slowly. The resulting slurry was filtered through a Celite pad, and the filtrate was extracted twice with EA (200 mL). The combined organic layers were dried over anhydrous -56-

[0347] FH13103096.12 Attorney Docket No.: LCH-03225

[0348] Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 2) to obtain the title compound as a colorless oil (5.10 g, 97%).

[0349] MS m / z: 173 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.33 (dd, 7 = 8.0, 1.3 Hz, 1H), 7.28 (s, 1H), 7.00 (d, 7 = 2.1 Hz, 1H), 6.87 (dd, 7 = 8.0, 1.9 Hz, 1H), 4.68 (d, 7 = 5.9 Hz, 2H), 3.92 (d, 7 = 1.1 Hz, 3H).

[0350] (Step 2) (2-bromo-4-chloro-5-methoxyphenyl)methanol

[0351] The title compound (4.30 g, 58%) was synthesized following the procedure described in preparative example A-5 using the compound (5.10 g, 29.5 mmol) obtained in (step 1) above.

[0352] MS m / z: 252 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.53 (s, 1H), 7.12 (s, 1H), 4.72 (dd, 7 = 6.1, 0.8 Hz, 2H), 3.92 (s, 3H), 1.97 (t, 7 = 6.1 Hz, 1H).

[0353] (Step 3) 6-chloro-5-methoxyisobenzofuran-l(3H)-one (Int. A-8)

[0354] The title compound (2.60 g, 62%) was synthesized following the procedure described in preparative example A-5 (step 2) using the compound (4.30 g, 17.1 mmol) obtained in (step 2) above.

[0355] MS m / z: 199 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.91 (s, 1H), 6.97 (s, 1H), 5.26 (d, 7 = 1.0 Hz, 2H), 4.01 (s, 3H).

[0356] (Preparative example A-91 5-bromo-6-fluoro-3H-isobenzofuran- 1-one (Int. A-9)

[0357] To a solution of 4-bromo-5-fluoro-2-methyl-benzoic acid (20 g, 85.8 mmol) in EA (200 mL) and water (200 mL) were added sodium bromate (38.9 g, 257 mmol) and NaHSCh (26.8 g, 257 mmol). The mixture was stirred at 20 °C for 48 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (10 mM NH4HCO3 in ACN : distilled water = 2 : 8 to 6 : 4) to obtain the title compound as a white solid (11.8 mg, 60%).

[0358] 1H NMR (400 MHz, CDCh) 5 7.88 (d, J = 5.38 Hz, 1 H), 7.63 (d, J = 6.50 Hz, 1 H), 5.30 (s, 2 H).

[0359] (Preparative example A- 101 6-fluoro-5-isopropoxyisobenzofuran-l(3H)-one (Int. A-10)

[0360] -57-

[0361] FH13103096.12 Attorney Docket No.: LCH-03225

[0362] (Step 1) methyl 4-fluoro-3-isopropoxybenzoate

[0363] A mixture of methyl 4-fluoro-3-hydroxybenzoate (1.00 g, 5.87 mmol) and 2- bromopropane (1.45 g, 11.8 mmol) in DMF (20 mL) was stirred at 60 °C for 20 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (DCM : n-hexane = 1 : 1) to obtain the title compound as a colorless oil (1.20 g, 96 %).

[0364] MS m / z: 213 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.73 - 7.54 (m, 2H), 7.11 (td, J = 9.7, 4.8 Hz, 1H), 4.63 (q, J = 6.1 Hz, 1H), 3.91 (dd, J = 3.5, 2.0 Hz, 3H), 1.44 - 1.32 (m, 6H).

[0365] (Step 2) (4-fluoro-3-isopropoxyphenyl)methanol

[0366] The title compound (400 mg, 92%) was synthesized following the procedure described in preparative example A-8 (step 1) using the compound (500 mg, 2.35 mmol) obtained in (step

[0367] 1) above.

[0368] MS m / z: 185 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.00 (qd, J = 9.4, 4.3 Hz, 2H), 6.90 - 6.74 (m, 1H), 4.55 (d, J = 5.4 Hz, 2H), 4.52 (d, J = 5.9 Hz, 1H), 2.66 (s, 1H), 1.34 (t, J = 6.1 Hz, 6H).

[0369] (Step 3) (2-bromo-4-fluoro-5 -isopropoxyphenyl)methanol

[0370] The title compound (235 mg, 41%) was synthesized following the procedure described in preparative example A-5 (step 2) using the compound (400 mg, 2.17 mmol) obtained in (step

[0371] 2) above.

[0372] MS m / z: 264 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.31 - 7.26 (m, 1H), 7.22 - 7.09 (m, 1H), 4.67 (d, J= 5.5 Hz, 2H), 4.56 (p, J = 5.9 Hz, 1H), 1.98 (q, J= 6.3 Hz, 1H), 1.42 - 1.30 (m, 6H).

[0373] (Step 4) 6-fluoro-5-isopropoxyisobenzofuran-l(3H)-one (Int. A-10)

[0374] The title compound (150 mg, 79%) was synthesized following the procedure described in preparative example A-5 (step 3) using the compound (235 mg, 0.89 mmol) obtained in (step

[0375] -58-

[0376] FH13103096.12 Attorney Docket No.: LCH-03225

[0377] 3) above.

[0378] MS m / z: 211 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.54 (dd, J = 9.6, 3.6 Hz, 1H), 7.02 (t, J = 5.6 Hz, 1H), 5.25 (d, J = 5.0 Hz, 2H), 4.78 - 4.63 (m, 1H), 1.44 (dt, 7 = 5.8, 2.9 Hz, 6H).

[0379] [Preparative example A- 11] 5-methoxy-6-nitroisobenzofuran-l(3H)-one (Int. A-ll)

[0380] (Step 1) methyl 3-methoxy-4-nitrobenzoate

[0381] To a solution of 3-hydroxy-4-nitrobenzoic acid (2.00 g, 10.9 mmol) in DMF (20 mL), were added K2CO3 (3.00 g, 21.8 mmol) and iodomethane (2.05 mL, 32.8 mmol) subsequently. After stirring at 70 °C for 16 h under nitrogen atmosphere, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 5 : 95) to obtain the title compound as a yellow solid (2.20 g, 94%).

[0382] MS m / z: 212 [M+l]+.1H NMR (400 MHz, CDCh) 57.89 - 7.81 (m, 1H), 7.76 (s, 1H), 7.73 - 7.63 (m, 1H), 4.11 - 4.00 (m, 3H), 3.97 (dd, J = 3.9, 1.6 Hz, 3H).

[0383] (Step 2) (3-methoxy-4-nitrophenyl)methanol

[0384] The title compound (1.40 g, 52%) was synthesized following the procedure described in preparative example A-8 (step 1) using the compound (3.00 g, 14.2 mmol) obtained in (step

[0385] 1) above.

[0386] MS m / z: 184 [M+l]+. ’ H NMR (400 MHz, CDCI3) 5 8.06 (q, 7 = 1.7 Hz, 1H), 7.37 (d, J = 4.3 Hz, 1H), 7.26 (s, 1H), 4.78 (d, 7 = 5.8 Hz, 2H), 4.00 (dd, J = 3.4, 1.8 Hz, 3H), 2.19 - 1.98 (m, 1H).

[0387] (Step 3) (2-bromo-5 -methoxy-4-nitrophenyl)methanol

[0388] The title compound (1.10 g, 57%) was synthesized following the procedure described in preparative example A-5 (step 2) using the compound (1.37 g, 7.47 mmol) obtained in (step

[0389] 2) above.

[0390] MS m / z: 264 [M+2]+

[0391] FH13103096.12 Attorney Docket No.: LCH-03225

[0392] (Step 4) 5-methoxy-6-nitroisobenzofuran-l(3H)-one (Int. A-ll)

[0393] A mixture of the compound prepared in (step 3) above (1.11 g, 4.23 mmol) and CuCN (758 mg, 8.47 mmol) dissolved in DMF (8 mL) was stirred at 110 °C for 18 h under carbon monoxide atmosphere (1 atm). On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as a yellow solid (230 mg, 26%).

[0394] MS m / z: 210 [M+l]+. ’ H NMR (400 MHz, CDC13) 5 8.32 (s, 1H), 7.15 (s, 1H), 5.35 (d, J = 1.0 Hz, 2H), 4.07 (s, 3H).

[0395] [Preparative example A- 121 6-methoxy-3-oxo-l,3-dihydroisobenzofuran-5-carbonitrile (Int.

[0396] A-12)

[0397] (Step 1) 6-amino-5-methoxyisobenzofuran- l(3H)-one

[0398] To a solution of 5-methoxy-6-nitroisobenzofuran-l(3H)-one (500 mg, 2.39 mmol) in EA (30 mL) was added Pd / C (100 mg), and the mixture was stirred at RT for 14 h under hydrogen atmosphere (1 atm). On completion, the reaction mixture was filtered through a Celite pad, then the filtrate was concentrated under reduced pressure to obtain the title compound as a yellow solid (423 mg, 99%), which was used for the next step without further purification.

[0399] MS m / z: 180 [M+l]+.1H NMR (400 MHz, DMSO-O 57.04 (s, 1H), 6.95 (s, 1H), 5.22 - 5.11 (m, 4H), 3.88 (s, 3H).

[0400] (Step 2) 6-bromo-5-methoxyisobenzofuran-l(3H)-one

[0401] The compound prepared in (step 1) above (423 mg, 2.36 mmol) was dissolved in 48 wt% HBr aqueous solution (7 mL), and NaNCL (179 mg, 2.60 mmol) in water (3.5 mL) was added thereto at 0 °C. The mixture was stirred at 0 °C for additional 0.5 h under nitrogen atmosphere, then CuBr (406 mg, 2.83 mmol) dissolved in 48 wt% HBr aqueous solution (3.50 mL) was added dropwise thereto. The reaction mixture was stirred at 80 °C for 1 h under nitrogen atmosphere. After being cooled to RT, the reaction was quenched by adding saturated -60-

[0402] FH13103096.12 Attorney Docket No.: LCH-03225

[0403] NaHCCL aqueous solution (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n- hexane = 1 : 1) to obtain the title compound as a beige solid (243 mg, 42%).

[0404] MS m / z: 243 [M]+.1H NMR (400 MHz, CDC13) 5 8.09 (s, 1H), 6.93 (s, 1H), 5.24 (s, 2H), 4.00 (s, 3H).

[0405] (Step 3) 6-methoxy-3-oxo-l,3-dihydroisobenzofuran-5-carbonitrile (Int. A-12)

[0406] The compound prepared in (step 2) above (100 mg, 0.411 mmol) was dissolved in DMF (3 mL), then Pd(PPht)4 (47.5 mg, 0.0411 mmol) and Zn(CN)2 (58.0 mg, 0.494 mmol) were added thereto. The reaction mixture was stirred at 130 °C for 4 h under nitrogen atmosphere. After being cooled to RT, the solvent was removed under reduced pressure. The resulting residue was diluted with distilled water (10 mL), then extracted twice with EA (10 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 7 : 3) to obtain the title compound as a yellow solid (69.6 mg, 89%).

[0407] MS m / z: 190 [M+l]+. ’ H NMR (400 MHz, CDC13) 5 8.13 (s, 1H), 7.04 (s, 1H), 5.33 (s, 2H), 4.06 (s, 3H).

[0408] (Preparative example A- 131 5-fluoro-6-methoxyisobenzofuran-l(3H)-one (Int. A-13)

[0409] The title compound (67.7 mg, 13%) was synthesized in the same manner as described in preparative example A-l (step 1) using 4-fluoro-3-methoxybenzoic acid (500 mg, 2.94 mmol).

[0410] MS m / z: 183 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.45 (d, J = 7.5 Hz, 1H), 7.19 (d, J = 9.5 Hz, 1H), 5.24 (s, 2H), 3.96 (d, J = 1.9 Hz, 3H).

[0411] (Preparative example A- 141 3-fluoro-2-methyl-7H-furo[3,4-b]pyridin-5-one (Int. A-14)

[0412] (Step 1) methyl 2-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-fluoro-6-methyl-pyridine- 3 -carboxylate

[0413] To a mixture of methyl 2-chloro-5-fluoro-6-methyl-pyridine-3-carboxylate (1.00 g, 4.91 mmol) and tert-butyl-dimethyl-(tributylstannylmethoxy)silane (3.21 g, 7.37 mmol) in 1,4- -61-

[0414] FH13103096.12 Attorney Docket No.: LCH-03225 dioxane (20 mL) was added XPhos Pd G2 (193 mg, 245 pmol). After stirring at 80 °C for 12 h under nitrogen atmosphere, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The resulting residue was purified by prep-HPLC (10 mM NH4HCO3 in ACN : distilled water = 6 : 4 to 1 : 0) to obtain the title compound as a colorless oil (1.90 g, 61%).

[0415] ‘ H NMR (400 MHz, CDCh) 5 7.74 (d, J = 9.26 Hz, 1H), 5.06 (s, 2H), 3.91 (s, 3H), 2.56 (d, J = 2.88 Hz, 3H), 1.01 - 0.75 (m, 9H), 0.07 (s, 6H).

[0416] (Step 2) methyl 5-fluoro-2-(hydroxymethyl)-6-methyl-pyridine-3-carboxylate

[0417] A solution of the compound prepared in (step 1) above (1.90 g, 6.06 mmol) in AcOH / THF / H2O (10 mL, 3 : 1 : 1) was stirred at RT for 12 h under nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by prep-HPLC (10 mM N^HCChin ACN : distilled water = 6 : 4 to 1 : 0) to obtain the title compound as a white solid (780 mg, 65%).

[0418] ’ H NMR (400 MHz, CDCh) 5 7.98 (d, J = 9.26 Hz, 1H), 5.01 (d, J = 1.25 Hz, 2H), 3.94 (s, 3H), 2.62 (d, J = 2.75 Hz, 3H).

[0419] (Step 3) 3-fluoro-2-methyl-7H-furo[3,4-b]pyridin-5-one_(Int. A-14)

[0420] To a mixture of compound prepared in (step 2) above (780 mg, 3.92 mmol) in DCM / H2O (8 mL, 1 : 1) was added K2CO3 (24 mg, 1.76 mmol). After stirring at 0 °C for 2 h under nitrogen atmosphere, the reaction mixture was diluted with distilled water (10 mL), then extracted twice with DCM (20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a white solid (500 mg, 76%), which was used for the next step without further purification.

[0421] 1H NMR (400 MHz, DMSO-rfe) 5 8.20 (d, J = 8.4 Hz, 1H), 5.38 (s, 2H), 2.60 (d, J = 3.3 Hz, 3H).

[0422] [Preparative example A- 15] 3-methoxy-7H-furo[3,4-b]pyridin-5-one (Int. A-15)

[0423] -62-

[0424] FH13103096.12 Attorney Docket No.: LCH-03225

[0425] (Step 1) 3-bromo-5-methoxv-pvridine-2-carboxylic acid

[0426] A mixture of 3-bromo-5-methoxy-pyridine-2-carbonitrile (10.0 g, 46.9 mmol) dissolved in 2 N NaOH aqueous solution (115 mL) was stirred at 100 °C for 1 h under nitrogen atmosphere. On completion, the aqueous layer was neutralized to pH 2~3 with 1 N HC1 aqueous solution. The resulting precipitate was filtered, washed with distilled water, then dried in vacuo to obtain the title compound as a white solid (10.0 g, 92%), which was used for the next step without further purification.

[0427] ’ H NMR (400 MHz, DMSO-tfc) 5 13.86 - 12.74 (m, 1H), 8.31 (d, J = 2.3 Hz, 1H), 7.80 (d, J = 2.3 Hz, 1H), 3.90 (s, 3H).

[0428] (Step 2) isobutoxycarbonyl 3-bromo-5-methoxy-pyridine-2-carboxylate

[0429] A mixture of the compound prepared in (step 1) above (10.0 g, 43.1 mmol) and isobutyl chloroformate (8.46 mL, 64.7 mmol), 4-methylmorpholine (6.54 g, 64.65 mmol) in DME (100 mL) was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction mixture was filtered to remove the precipitate. The filtrate was concentrated under reduced pressure to obtain the title compound as a yellow oil (14.0 g), which was used for the next step without further purification.

[0430] (Step 3) (3-bromo-5-methoxv-2-pvridyl)methanol

[0431] To a solution of the compound prepared in (step 2) above (10.0g, 30.1 mmol) in MeOH (50 mL) was added NaBH4 (2.62 g, 69.3 mmol) at 0 °C under nitrogen atmosphere. The mixture was stirred at RT for 1 h under nitrogen atmosphere. On completion, the residue was diluted with distilled water (60 mL), then extracted twice with 10% MeOH in DCM (90 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 0 : 1 to 3 : 7) to obtain the tide compound as a white solid (4.0 g, 61%).

[0432] 1H NMR (400 MHz, CDC13) 5 8.24 (d, J = 2.5 Hz, 1H), 7.45 (d, J = 2.5 Hz, 1H), 4.71 (s, 2H), 3.89 (s, 3H).

[0433] (Step 4) 3-methoxv-7H-furol3.4-b1pvridin-5-one (Int. A-15)

[0434] The title compound (3.00 g, 94%) was synthesized following the procedure described

[0435] FH13103096.12 Attorney Docket No.: LCH-03225 in preparative example A-5 (step 3) using the compound (4.0 g, 18.3 mmol) obtained in (step 3) above.

[0436] 1H NMR (400 MHz, DMSO-&) 5 8.64 (d, J = 2.8 Hz, 1H), 7.82 (d, J = 2.8 Hz, 1H), 5.36 (s, 2H), 3.92 (s, 3H).

[0437] [Preparative example A- 161 6-fluoro-5-((4-methoxvbenzyl)oxv)isobenzofuran- 1 (3H)-one

[0438] (Int. A- 16)

[0439] (Step 1) 6-fluoro-5-hydroxyisobenzofuran-l(3H)-one

[0440] The title compound (760 mg, 82%) was synthesized following the procedure described in preparative example A-l (step 2) using Int. A-5 (1.00 g, 5.49 mmol).

[0441] MS m / z: 169 [M+l]+. ’ H NMR (400 MHz, DMSO-tfe) 5 7.59 (d, J = 9.8 Hz, 1H), 7.14 (d, J = 7.0 Hz, 1H), 5.26 (s, 2H).

[0442] (Step 2) 6-fluoro-5-((4-methoxybenzyl)oxy)isobenzofuran-l(3H)-one (Int. A-16)

[0443] To a solution of the compound prepared in (step 1) above (400 mg, 2.38 mmol) and 4- methoxybenzylalcohol (0.60 mL, 4.76 mmol) in THF (5 mL) were added PPht (1.20 g, 1.99 mmol) and DIAD (0.95 mL, 4.84 mmol) at 0 °C. The reaction mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (100 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 3 : 7) to obtain the title compound as a yellow solid (498 mg, 72%).

[0444] MS m / z: 289 [M+l]+

[0445] [Preparative example A- 171 6-chloro-5-((4-methoxybenzyl)oxy)isobenzofuran-l(3H)-one (Int. A- 17)

[0446] The title compound was synthesized in the same manner as described in preparative example A-16 using Int. A-8 instead of Int. A-5.

[0447] MS m / z: 305 [M+l]+

[0448] [Preparative example A- 181 2-bromo-4-methoxy-5 -((4-methoxybenzyl)oxy) benzaldehyde

[0449] -64-

[0450] FH13103096.12 Attorney Docket No.: LCH-03225

[0451] (Int. A- 18)

[0452] The title compound (920 mg, 60%) was synthesized following the procedure described in preparative example A- 16 (step 2) using 2-bromo-5-hydroxy-4-methoxybenzaldehyde (1.00 g, 4.32 mmol).

[0453] MS m / z: 352 [M+l]+

[0454] (Preparative example A- 191 2-bromo-4-fluoro-5-(trifluoromethyl)benzaldehyde (Int. A-19)

[0455] (Step 1) 5-fluoro-2-iodo-4-(trifluoromethvl)aniline

[0456] To a solution of 3-fluoro-4-(trifluoromethyl)aniline (4.40 g, 24.6 mmol) in acetic acid (24 mL) was added NIS (5.53 g, 24.6 mmol) portionwise with stirring. The reaction mixture was stirred at RT for 25 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (120 mL), then extracted twice with EA (120 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 1 : 3) to obtain the tide compound as a brown oil (6.75 g, 90%).

[0457] MS m / z: 306 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.82 - 7.74 (m, 1H), 6.53 - 6.45 (m, 1H), 4.53 (s, 2H).

[0458] (Step 2) 2-amino-4-fluoro-5 -(trifluoromethyl)benzonitrile

[0459] The title compound (4.07 g, 90%) was synthesized following the procedure described in preparative example A- 12 (step 3) using the compound (6.75 g, 22.1 mmol) obtained in (step 1) above.

[0460] MS m / z: 205 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.66 (d, J = 7.3 Hz, 1H), 6.53 (d, J = 11.5 Hz, 1H), 4.87 (s, 2H).

[0461] (Step 3) 2-bromo-4-fluoro-5 -(trifluoromethyl)benzonitrile

[0462] To a solution of the compound prepared in (step 2) above (300 mg, 1.47 mmol) in ACN (5 mL) were added CuBr (394 mg, 1.76 mmol) and tert-butyl nitrite (0.291 mL, 2.20 mmol) at 0 °C. After stirring at RT for 3 h under nitrogen atmosphere, the reaction was quenched by -65-

[0463] FH13103096.12 Attorney Docket No.: LCH-03225 adding 1 N HC1 aqueous solution (60 mL), then extracted twice with EA (60 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 9) to obtain the title compound as an orange oil (224 mg).

[0464] MS m / z: 269 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.92 (d, J = 6.9 Hz, 1H), 7.62 (d, J = 9.3 Hz, 1H).

[0465] (Step 4) 2-bromo-4-fluoro-5-(trifluoromethyl)benzaldehyde (Int. A-19)

[0466] To a solution of the compound prepared in (step 3) above (224 mg, 0.84 mmol) in DCM (4 mL), was added 1 M DIBAL-H solution in toluene (0.90 mL, 0.90 mmol) dropwise at 0 °C. After stirring at RT for 1 h under nitrogen atmosphere, the reaction was quenched by adding 1 N HC1 aqueous solution (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 4) to obtain the title compound as a colorless oil (82.6 mg)

[0467] MS m / z: 272 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 10.30 (s, 1H), 8.22 (d, J = 7.6 Hz, 1H), 7.57 (d, J = 9.3 Hz, 1H).

[0468] [Preparative example A-20] 4-bromo-2-fluoro-5 -formylbenzonitrile (Int. A-20)

[0469] (Step 1) methyl 2-amino-4-fluoro-5-iodobenzoate

[0470] The title compound (5.48 g, 63%) was synthesized following the procedure described in preparative example A-19 (step 1) using methyl 2-amino-4-fluorobenzoate (5.00 g, 29.6 mmol).

[0471] MS m / z: 296 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 8.22 (d, . / = 7.1 Hz, 1H), 6.39 (d, J = 9.8 Hz, 1H), 5.92 (s, 2H), 3.87 (s, 3H).

[0472] (Step 2) methyl 2-amino-5-cyano-4-fluorobenzoate

[0473] The title compound (3.11 g, 86%) was synthesized following the procedure described in preparative example A-12 (step 3) using the compound (5.48 g, 18.6 mmol) obtained in (step 1) above.

[0474] MS m / z: 195 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 8.19 (d, J = 7.4 Hz, 1H), 6.40

[0475] -66-

[0476] FH13103096.12 Attorney Docket No.: LCH-03225

[0477] (d, J = 11.0 Hz, 1H), 3.90 (s, 3H).

[0478] (Step 3) methyl 2-bromo-5-cyano-4-fluorobenzoate

[0479] The title compound (3.89 g, 94%) was synthesized following the procedure described in preparative example A-19 (step 3) using the compound (3.11 g, 16.0 mmol) obtained in (step 2) above.

[0480] MS m / z: 258 [M]+.1H NMR (400 MHz, CDCh) 58.17 (d, J = 6.8 Hz, 1H), 7.61 (d, J = 8.3 Hz, 1H), 3.97 (s, 3H).

[0481] (Step 4) 4-bromo-2-fluoro-5-(hvdroxvmethyl)benzonitrile

[0482] To a solution of the compound prepared in (step 3) above (3.89 g, 15.1 mmol) in THF (75 mL) was added LiBH4 (328 mg, 15.1 mmol) portionwise at 0 °C. After stirring at RT for 4 h under nitrogen atmosphere, the reaction was quenched by adding distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 3 : 7) to obtain the title compound as a white solid (2.75 g, 79%).

[0483] MS m / z: 230 [M]+.1H NMR (400 MHz, CDCh) 57.82 (d, J = 6.8 Hz, 1H), 7.47 (d, J = 8.1 Hz, 1H), 4.74 (d, J = 5.8 Hz, 2H), 2.06 (t, J = 5.1 Hz, 1H).

[0484] (Step 5) 4-bromo-2-fluoro-5 -formylbenzonitrile (Int. A-20)

[0485] To a solution of the compound prepared in (step 4) above (1.50 g, 6.52 mmol) in DCM (33 mL) was added Dess-Martin periodinane (3.04 g, 7.17 mmol) carefully at 0 °C with stirring. After stirring at RT for 2 h under nitrogen atmosphere, the reaction was quenched by adding 1 N HC1 aqueous solution (50 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n- hexane = 1 : 4) to obtain the tide compound as a white solid (1.20 g, 76%).

[0486] MS m / z: 228 [M]+. ’ H NMR (400 MHz, CDCh) 5 10.27 (s, 1H), 8.22 (d, . / = 6.9 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H).

[0487] [Preparative example A-211 4-bromo-5-formyl-2-((4-methoxybenzyl)oxy) benzonitrile (Int.

[0488] A-21)

[0489] FH13103096.12 Attorney Docket No.: LCH-03225

[0490] (Step 1) methyl 2-amino-4-fluoro-5-iodobenzoate

[0491] The title compound (798 mg, 46%) was synthesized following the procedure described in preparative example A-19 (step 1) using methyl 2-amino-4- fluorobenzoate (1.00 g, 5.91 mmol).

[0492] MS m / z: 296 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 8.21 (d, J = 7.3 Hz, 1H), 6.39 (d, J = 9.8 Hz, 1H), 5.92 (s, 2H), 3.86 (s, 3H).

[0493] (Step 2) methyl 2-amino-5-cyano-4-fluorobenzoate

[0494] The title compound (471 mg, 90%) was synthesized following the procedure described in preparative example A-12 (step 3) using the compound (798 mg, 2.70 mmol) obtained in (step 1) above.

[0495] MS m / z: 195 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 8.19 (d, J = 7.4 Hz, 1H), 6.39 (d, J = 11.0 Hz, 1H), 3.89 (s, 3H).

[0496] (Step 3) 2-amino-5-cyano-4- methoxybenzoic acid

[0497] A mixture of the compound prepared in (step 2) above (100 mg, 0.515 mmol) and CS2CO3 (503 mg, 1.55 mmol) dissolved in MeOH (1.5 mL) was stirred at 80 °C for 4 h under nitrogen atmosphere. The reaction was quenched by adding 1 N HC1 aqueous solution (20 mL), then extracted twice with DCM (20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a white solid (81.8 mg, 83%), which was used for the next step without further purification.

[0498] MS m / z: 193 [M+l]+. ’ H NMR (400 MHz, DMSO-O 5 12.77 (s, 1H), 7.93 (s, 1H), 7.52 (s, 2H), 6.39 (s, 1H), 3.83 (s, 3H).

[0499] (Step 4) methyl 2-amino-5-cyano-4-methoxybenzoate

[0500] The title compound (76.3 mg, 87%) was synthesized following the procedure described in preparative example A- 11 (step 1) using the compound (81.8 mg, 0.43 mmol) obtained in (step 3) above.

[0501] -68-

[0502] FH13103096.12 Attorney Docket No.: LCH-03225

[0503] MS m / z: 207 [M+l]+. ’ H NMR (400 MHz, CDCI3) 5 8.12 (s, 1H), 6.35 (s, 2H), 6.06 (s, 1H), 3.89 (s, 3H), 3.86 (s, 3H).

[0504] (Step 5) methyl 2-bromo-5-cyano-4-methoxybenzoate

[0505] The title compound (86.1 mg, 86%) was synthesized following the procedure described in preparative example A-19 (step 3) using the compound (76.3 mg, 0.37 mmol) obtained in (step 4).

[0506] MS m / z: 271 [M+l]+. ’ H NMR (400 MHz, CDCI3) 5 8.15 (s, 1H), 7.28 (s, 1H), 4.00 (s, 3H), 3.93 (s, 3H).

[0507] (Step 6) methyl 2-bromo-5-cyano-4-hydroxybenzoate

[0508] A mixture of the compound prepared in (step 5) above (35.0 mg, 0.130 mmol) and LiCl (27.5 mg, 0.648 mmol) dissolved in DMF (1 mL) was stirred at 120 °C for 22 h under nitrogen atmosphere. After being cooled to RT, the reaction was quenched by adding 1 N HC1 aqueous solution (10 mL), then extracted twice with 10% MeOH in DCM (10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 15) to obtain the title compound as a light brown solid (21.7 mg, 65%).

[0509] MS m / z: 257 [M+l]+. ’ H NMR (400 MHz, CDCI3) 5 8.12 (s, 1H), 7.35 (s, 1H), 3.93 (s, 3H).

[0510] (Step 7) methyl 2-bromo-5-cyano-4-((4-methoxybenzyl)oxy)benzoate

[0511] A mixture of the compound prepared in (step 6) above (294 mg, 1.15 mmol), K2CO3 (476 mg, 3.44 mmol), 4-methoxybenzyl chloride (0.171 mL, 1.26 mmol), and tetra-n- butylammonium iodide (42.4 mg, 0.115 mmol) dissolved in acetone (10 mL) was stirred at RT for 24 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (40 mL), then extracted twice with DCM (40 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 5) to obtain the title compound as a white solid (333 mg, 77%).

[0512] MS m / z: 377 [M+l]+. ’ H NMR (400 MHz, CDCI3) 5 8.15 (s, 1H), 7.38 (s, 1H), 7.35 (d, J = 5.6 Hz, 2H), 6.94 (d, J = 8.8 Hz, 2H), 5.18 (s, 2H), 3.92 (s, 3H), 3.83 (s, 3H).

[0513] (Step 8) 4-bromo-5-(hvdroxvmethvl)-2-((4-methoxvbenzyl)oxv)benzonitrile

[0514] The title compound (22.6 mg, 88%) was synthesized following the procedure described in preparative example A- 8 (step 1) using the compound (27.7 mg, 0.07 mmol) obtained in

[0515] -69-

[0516] FH13103096.12 Attorney Docket No.: LCH-03225

[0517] (step 7).

[0518] MS m / z: 349 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.70 (s, 1H), 7.37 (d, J = 8.8 Hz, 2H), 7.24 (s, 1H), 6.93 (d, J = 8.6 Hz, 2H), 5.13 (s, 2H), 4.69 (d, J = 5.8 Hz, 2H), 3.82 (s, 3H), 1.93 (t, 7 = 5.9 Hz, 1H).

[0519] (Step 9) 4-bromo-5-formyl-2-((4-methoxybenzyl)oxy)benzonitrile (Int. A-21)

[0520] The title compound (274 mg, 84%) was synthesized following the procedure described in preparative example A-20 (step 5) using the compound (328 mg, 0.94 mmol) obtained in (step 8).

[0521] MS m / z: 347 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 10.18 (s, 1H), 8.15 (s, 1H), 7.37 (d, J = 8.8 Hz, 2H), 7.29 (s, 1H), 6.95 (d, J = 8.9 Hz, 2H), 5.22 (s, 2H), 3.83 (s, 3H).

[0522] Preparation Example 2: Synthesis of intermediate B series

[0523] [Preparative example B-l] 3-fluoro-2-methoxy-[l,3]dioxolo[4',5':5,6]indeno[l,2- c]isochromene-5, 12-dione (Int. B-l)

[0524] (Step 1) 3-bromo-6-fluoro-5-methoxyisobenzofuran-l(3H)-one

[0525] To a mixture of Int. A-5 (133 mg, 0.730 mmol) and benzoyl peroxide (17.7 mg, 0.073 mmol) dissolved in CCh (7.3 mL) was added NBS (143 mg, 0.804 mmol) portionwise with stirring. The reaction mixture was stirred at 80 °C for 3 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a white solid (79.5 mg, 41%).

[0526] MS m / z: 262 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.55 (d, J = 8.9 Hz, 1H), 7.31 (s, 1H), 7.10 (d, J = 6.8 Hz, 1H), 4.03 (s, 3H).

[0527] (Step 2) 6-fluoro-3-hydroxy-5-methoxyisobenzofuran-l(3H)-one

[0528] A solution of the compound prepared in (step 1) above (200 mg, 0.766 mmol) in distilled water (7.7 mL) was heated at 100 °C for 5 h. On completion, the reaction mixture was concentrated under reduced pressure. DCM (10 mL) was added to the remaining residue, and the resulting precipitate was filtered, washed with DCM, then dried in vacuo to obtain the title compound as a white solid (151 mg, 99%).

[0529] -70-

[0530] FH13103096.12 Attorney Docket No.: LCH-03225

[0531] MS m / z: 199 [M+l]+. ’ H NMR (400 MHz, DMSO-tfe) 5 8.21 (d, J = 8.3 Hz, 1H), 7.68 (d, J = 9.6 Hz, 1H), 7.44 (d, J = 7.3 Hz, 1H), 6.59 (d, J = 8.1 Hz, 1H), 3.98 (s, 3H).

[0532] (Step 3) 3-fluoro-2-methoxy-[l,3]dioxolo[4',5':5,6]indeno[l,2-c]isochromene-5, 12- dione (Int. B-l)

[0533] To a mixture of Int. A-2 (98.9 mg, 0.555 mmol) and the compound prepared in (step 2) above (100 mg, 0.505 mmol) dissolved in EA (2 mL) was added NaOMe which was freshly prepared from sodium metal (98.7 mg, 4.29 mmol) and MeOH (4 mL) at RT. The reaction mixture was stirred at 80 °C for 24 h under nitrogen atmosphere. After being cooled to RT, the reaction was quenched by adding 1 N HC1 aqueous solution to adjust pH below 2, then concentrated under reduced pressure. Acetic anhydride (2 mL) was added to the remaining residue, then the mixture was stirred at 100 °C for 12 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (10 mL), then extracted twice with DCM (10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the title compound as a purple solid (145 mg, 85%).

[0534] MS m / z: 341 [M+l]+

[0535] The following intermediates (Int.B-2 ~ Int. B-5) were synthesized in the same manner as described in preparative example B-l using Int. A-2 and the corresponding isobenzofuranones instead of Int. A-5.

[0536] [Preparative example B-21 3-chloro-2-methoxy-[l,3]dioxolo[4',5':5,6]indeno[l,2- c]isochromene-5, 12-dione (Int. B-2)

[0537] [Preparative example B-31 3-fluoro-2-isopropoxy- [ 1 ,3]dioxolo [4' ,5 ' :5 ,6]indeno [1,2- c]isochromene-5, 12-dione (Int. B-3)

[0538] [Preparative example B-41 2-fluoro-3-methoxy-[l,3]dioxolo[4',5':5,6]indeno[l,2- c]isochromene-5, 12-dione (Int. B-4)

[0539] [Preparative example B-51 l-fluoro-2-methoxy-[l,3]dioxolo[4',5':5,6]indeno[l,2- c]isochromene-5, 12-dione (Int. B-5)

[0540] FH13103096.12 Attorney Docket No.: LCH-03225

[0541] [Preparative example B-61 2-methoxy-3-nitro-[l,3]dioxolo[4',5':5,6]indeno[l,2- c]isochromene-5, 12-dione (Int. B-6)

[0542] (Step 1) 2-(hydroxymethyl)-4-methoxy-5-nitrobenzoic acid

[0543] To a solution of Int. A-ll (100 mg, 0.47 mmol) in THF (2 mL) was added NaOH (57.0 mg, 1.43 mmol) dissolved in distilled water (1.5 mL). The reaction mixture was stirred at RT for 1 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (10 mL), then extracted twice with EA (10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (MeOH : DCM = 5 : 95) to obtain the title compound as a brown solid (86.0 mg, 79%).

[0544] MS m / z: 228 [M+l]+.1H NMR (400 MHz, CD3OD) 5 8.48 (s, 1H), 7.66 (s, 1H), 5.04 (d, J = 1.0 Hz, 2H), 4.05 (s, 3H).

[0545] (Step 2) 3-hydroxy-5 -methoxy-6-nitroisobenzofuran- 1 (3H)-one

[0546] To a solution of the compound prepared in (step 1) above (86.0 mg, 0.379 mmol) in DCM (3 mL) was added MnCh (329 mg, 3.79 mmol). The reaction mixture was stirred at RT for 17 h under nitrogen atmosphere. On completion, the reaction mixture was filtered through a Celite pad, washed with EA, then the filtrate was concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 5 : 95) to obtain the title compound as a yellow. (35.0 mg, 41%).

[0547] -72-

[0548] FH13103096.12 Attorney Docket No.: LCH-03225

[0549] MS m / z: 226 [M+l]+. ’ H NMR (400 MHz, CDCI3) 5 8.26 (s, 1H), 7.31 (s, 1H), 6.63 (s, 1H), 4.09 (s, 3H).

[0550] (Step 3) 2-methoxy-3-nitro-[l,3]dioxolo[4',5':5,6]indeno[l,2-c]isochromene-5, 12- dione (Int. B-6)

[0551] The title compound (15.0 mg, 26%) was synthesized following the procedure described in preparative example B-l (step 3) using the compound (35.0 mg, 0.150 mmol) obtained in (step 2) above.

[0552] MS m / z: 368 [M+l]+. ’ H NMR (400 MHz, CDCI3) 5 8.74 (s, 1H), 7.85 (s, 1H), 7.12 (s, 1H), 7.01 (s, 1H), 6.14 (s, 2H), 4.13 (s, 3H).

[0553] The following intermediates, Int.B-7~ Int. B-9 were synthesized in the same manner as described in preparative example B-6 using the corresponding isobenzofuranones instead of Int. A-ll.

[0554] (Preparative example B-71 2-methoxy-5 , 12-dioxo-5 , 12-dihydro- [ 1 ,3]dioxolo

[0555] [4',5':5,6]indeno[l,2-c]isochromene-3-carbonitrile (Int. B-7)

[0556] (Preparative example B-81 3-fluoro-2-((4-methoxybenzyl)oxy)-[l ,3]dioxolo [4', 5':5,6]indeno[l,2-c]isochromene-5, 12-dione (Int. B-8)

[0557] (Preparative example B-91 3-chloro-2-((4-methoxybenzyl)oxy)-[l,3]dioxolo[4',5':5,6] indeno[l,2-c]isochromene-5, 12-dione (Int. B-9)

[0558] (Preparative example B-101 2-hydroxy-3-methoxy- [ 1 ,3]dioxolo [4',5 ' :5 ,6]indeno [1,2- c]isochromene-5, 12-dione (Int. B-10)

[0559] FH13103096.12 Attorney Docket No.: LCH-03225

[0560] (Step 1) 3-hydroxy-6-methoxy-5-((4-methoxybenzyl)oxy)isobenzofuran-l(3H)-one

[0561] To a solution of Int. A-18 (920 mg, 2.62 mmol) in ACN / distilled water (26 mL, 6 : 1) were added Pd(OAc)2 (29.4 mg, 0.131 mmol), xantphos (151 mg, 0.262 mmol), and TEA (2.19 mL, 15.7 mmol). The reaction mixture was stirred at 80 °C for 16 h under carbon monoxide atmosphere (1 atm). After being cooled to RT, the reaction mixture was filtered through a Celite pad, washed with EA (100 mL). The filtrate was diluted with 1 N HC1 aqueous solution (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 9) to obtain the title compound as a white solid (820 mg, 99%).

[0562] MS m / z: 317 [M+l]+

[0563] (Step 2) 2-hydroxy-3-methoxy-[l,3]dioxolo[4',5':5,6]indeno[l,2-c]isochromene-5, 12- dione (Int. B-10)

[0564] The title compound (90.0 mg, 31%) was synthesized following the procedure described in preparative example B-l (step 3) using the compound (200 mg, 0.62 mmol) obtained in (step 1) above.

[0565] ’ ll NMR (400 MHz, DMSO-O 5 7.52 (s, 1H), 7.48 (s, 1H), 7.16 (d, J = 4.0 Hz, 2H), 6.18 (s, 2H), 3.87 (s, 3H).

[0566] The following intermediates (Int.B-11 ~ Int. B-14) were synthesized in the same manner as described in preparative example B-10 using the corresponding isobenzofuranones instead of Int. A- 18.

[0567] [Preparative example B-111 3-methoxy-2-(trifluoromethyl)-[l,3]dioxolo

[0568] [4', 5':5,6]indeno[l,2-c]isochromene-5, 12-dione (Int. B-ll)

[0569] [Preparative example B-12] 3-fluoro-2-(trifluoromethyl)-[l,3]dioxolo[4',5':5,6] indeno[l,2- c]isochromene-5, 12-dione (Int. B-12)

[0570] [Preparative example B-131 3-methoxy-5,12-dioxo-5,12-dihydro-[l,3]dioxolo [4’,5’:5,6] indeno[l,2-c]isochromene-2-carbonitrile (Int. B-13)

[0571] [Preparative example B-141 3-((4-methoxybenzyl)oxy)-5, 12-dioxo-5, 12-dihydro-

[0572] [l,3]dioxolo[4',5':5,6]indeno[l,2-c]isochromene-2-carbonitrile (Int. B-14)

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[0575] [Preparative example B-151 3-hydroxy-5 , 12-dioxo-5 , 12-dihydro- [ 1 ,3]dioxolo [4',5 ' :5 ,6] indeno[l,2-c]isochromene-2-carbonitrile (Int. B-15)

[0576] A mixture of Int. B-14 (66.3 mg, 0.15 mmol) and TFA (0.5 mL) dissolved in DCM (1 mL) was stirred at RT for 1 h under nitrogen atmosphere. On completion, the solvent was removed under reduced pressure, and the remaining residue was washed with DCM (10 mL), MeOH (2 mL), then n-hexane (20 mL) to obtain the tide compound as a brown solid (54.6 mg, 83%).

[0577] ’ H NMR (400 MHz, DMSO-rfc) 5 11.98 (s, 1H), 8.22 (s, 1H), 7.70 (s, 1H), 7.21 (d, J = 2.6 Hz, 2H), 6.19 (s, 2H).

[0578] [Preparative example B-161 3-fluoro-2-hydroxy- [ 1 ,3] dioxolo[4' ,5 ' :5 ,6]indeno [1,2- c]isochromene-5, 12-dione (Int. B-16)

[0579] The title compound (36.5 mg, 99%) was synthesized in the same manner as described

[0580] -75-

[0581] FH13103096.12 Attorney Docket No.: LCH-03225 in preparative example B- 15 using Int. B-8 (50.0 mg, 0.11 mmol) instead of Int.B-14.

[0582] MS m / z: 327 [M+l]+. ’ H NMR (400 MHz, DMSO-&) 57.83 (d, . / = 11.3 Hz, 1H), 7.67 (d, J = 8.3 Hz, 1H), 7.19 (d, J = 4.1 Hz, 2H), 6.19 (s, 2H).

[0583] [Preparative example B-17] 3-chloro-2-hydroxy-[l,3]dioxolo[4',5':5,6]indeno[l,2- c]isochromene-5, 12-dione (Int. B-17)

[0584] The title compound (64.1 mg, 87%) was synthesized in the same manner as described in preparative example B-15 using Int. B-9 (51.0 mg, 0.159 mmol) instead of Int.B-14.

[0585] MS m / z: 343 [M+l]+

[0586] [Preparative example B-181 3-fluoro-5,12-dioxo-5,12-dihydro-[l,3]dioxolo[4',5':5,6] indeno[l,2-c]isochromene-2-carbonitrile (Int. B-18)

[0587] (Step 1) 3-fluoro-5,12-dioxo-5,12-dihydro-[l,3]dioxolo[4',5':5,6]indeno[l,2- c]isochromen-2-yl trifluoromethanesulfonate

[0588] To a solution of Int. B-16 (100 mg, 0.301 mmol) in DCM (3 mL) were added pyridine (120 pL, 1.53 mmol) and trifluoromethanesulfonic anhydride (103 pL, 0.613 mmol) at -20 °C. The reaction mixture was stirred at 0 °C for 4 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (20 mL), then extracted twice with DCM (10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (DCM : n-hexane = 1 : 1) to obtain the title compound as a dark orange solid (67 mg, 48%).

[0589] MS m / z: 459 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 8.25 (d, J = 6.6 Hz, 1H), 8.12 (d, 7 = 9.4 Hz, 1H), 7.12 (s, 1H), 6.98 (s, 1H), 6.13 (s, 2H).

[0590] (Step 2) 3-fluoro-5,12-dioxo-5,12-dihydro-[l,3]dioxolo[4',5':5,6]indeno[l,2-c] isochromene-2-carbonitrile (Int. B-18)

[0591] To a solution of the compound prepared in (step 1) above (67.0 mg, 0.146 mmol) in DMF (1.5 mL) were added Pd(PPh3)4 (16.9 mg, 0.015 mmol) and Zn(CN)2 (20.6 mg, 0.175 -76-

[0592] FH13103096.12 Attorney Docket No.: LCH-03225 mmol). The reaction mixture was stirred at 130 °C for 4 h under nitrogen atmosphere. On completion, the mixture was concentrated under reduced pressure. Acetic anhydride (1.5 mL) was added to the remaining residue, then the mixture was stirred at 100 °C for 12 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (10 mL), then extracted twice with EA (10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the title compound brown solid (16.0 mg, 32%).

[0593] MS m / z: 336 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 8.60 (d, J = 6.1 Hz, 1H), 8.05 (d, J = 8.1 Hz, 1H), 7.13 (s, 1H), 6.99 (s, 1H), 6.14 (s, 2H).

[0594] (Preparative example B-191 3-chloro-5 , 12-dioxo-5 , 12-dihydro- [ 1 ,3]dioxolo

[0595] [4',5':5,6]indeno[l,2-c]isochromene-2-carbonitrile (Int. B-19)

[0596] The title compound was synthesized in the same manner as described in preparative example B-18 using Int. B-17 instead of Int. B-16.

[0597] MS m / z: 352 [M+l]+

[0598] (Preparative example B-20] 1 ,3-difluoro-2-methoxy-[l ,3]dioxolo[4',5':5,6] indeno[l ,2- c]isochromene-5, 12-dione (Int. B-20)

[0599] (Step 1) 3,5-difluoro-N,N-diisopropyl-4-methoxybenzamide

[0600] To a solution of 3, 5 -difluoro-4- methoxybenzoic acid (500 mg, 2.66 mmol) in DCM (9 mL) were added diisopropylamine (0.6 mL, 4.0 mmol), HATU (1.52 g, 3.99 mmol), and DIPEA (1.4 mL, 8.0 mmol) at 0 °C. The reaction mixture was stirred at RT for 19 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure.

[0601] -77-

[0602] FH13103096.12 Attorney Docket No.: LCH-03225

[0603] The residue was diluted with distilled water (60 ruL), then extracted twice with DCM (60 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 6) to obtain the title compound as a light brown oil (526 mg, 73%).

[0604] MS m / z: 272 [M+l]+.1H NMR (400 MHz, CDCh) 5 6.89 - 6.85 (m, 2H), 4.01 (s, 3H), 3.66 (s, 2H), 1.33 (s, 12H).

[0605] (Step 2) 3 ,5 -difluoro-2-formyl-N,N -diisopropyl -4-methoxybenzamide

[0606] The compound prepared in (step 1) above (526 mg, 1.94 mmol) was dissolved in THF (5 mL), and 2.5 M n-BuLi solution in n-hexane (0.9 mL, 2.78 mmol) was added thereto at - 70 °C. The reaction mixture was stirred at the same temperature for 1 h, then DMF (0.2 mL, 2.5 mmol) was added thereto. After stirring at RT for 2 h under nitrogen atmosphere, the reaction was quenched by adding saturated NH4CI aqueous solution (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 3) to obtain the title compound as a light brown oil (412 mg, 71%).

[0607] MS m / z: 300 [M+l]+.1H NMR (400 MHz, CDCh) 5 10.26 (d, J = 0.6 Hz, 1H), 6.81 - 6.75 (m, 1H), 4.05 (s, 3H), 3.55 - 3.46 (m, 2H), 1.58 (d, J = 6.9 Hz, 6H), 1.11 (d, J = 6.6 Hz, 6H).

[0608] (Step 3) 4,6-difluoro-3-hvdroxv-5-methoxvisobenzofuran-l(3H)-one

[0609] A solution of the compound prepared in (step 2) above (200 mg, 0.668 mmol) in a mixed solvent of acetic acid / 6 N HC1 aqueous solution (1.4 mL, 1 : 1) was stirred at 100 °C for 55 h under nitrogen atmosphere. After being cooled to RT, the reaction mixture was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 2 : 3) to obtain the title compound as a white solid (50.6 mg, 35%).

[0610] MS m / z: 217 [M+l]+. ’ H NMR (400 MHz, DMSO-tfe) 5 8.44 (s, 1H), 7.69 (d, J = 8.9 Hz, 1H), 6.95 (s, 1H), 4.06 (s, 3H).

[0611] (Step 4) l,3-difluoro-2-methoxy-[l,3]dioxolo[4',5':5,6]indeno[l,2-c]isochromene- 5, 12-dione (Int. B-20)

[0612] The title compound (97.0 mg) was synthesized following the procedure described in

[0613] -78-

[0614] FH13103096.12 Attorney Docket No.: LCH-03225 preparative example B-l (step 3) using the compound (50.6 mg, 0.23 mmol) obtained in (step 3) above.

[0615] MS m / z: 359 [M+l]+

[0616] The following intermediates (Int.B-21 ~ Int. B-23) were synthesized in the same manner as described in preparative example B-l using Int. A-5 and the corresponding isobenzofuranones instead of Int. A-2.

[0617] [Preparative example B-21] 3-fluoro-2,8,9-trimethoxyindeno[l,2-c]isochromene-5,ll-dione

[0618] (Int. B-21)

[0619] [Preparative example B-221 3 ,9 ,9-trifluoro-2-methoxy- [ 1 ,3]dioxolo [4',5 ' :5 ,6] indeno [1,2- c]isochromene-5, 12-dione (Int. B-22)

[0620] [Preparative example B-231 3-fluoro-2-methoxy- [ 1 ,3]dioxolo [4' ,5 ' :5 ,6]indeno [1,2- c]isochromene-5,12-dione-9,9-d2 (Int. B-23)

[0621] [Preparative example B-241 2-bromo-3-fluoro-7-methoxy-indeno[ 1 ,2-c]isochromene-5 ,11- dione (Int. B-24)

[0622] The title compound was synthesized in the same manner as described in preparative example B-l using Int. A-9 and 4-methoxy-3H-isobenzofuran-l-one.

[0623] FH13103096.12 Attorney Docket No.: LCH-03225

[0624] ’ H NMR (400 MHz, CDCI3) 5 8.41 (s, 1H), 8.15 - 7.97 (m, 1H), 7.71 - 7.45 (m, 1H), 7.41 - 7.12 (m, 2H), 3.96 (d, J = 16.3 Hz, 3H).

[0625] Preparation Example 3: Synthesis of intermediate C series

[0626] [Preparative example C-l] 2-(4-bromobutyl)isoindoline- 1,3-dione (Int. C-l)

[0627] To a solution of phthalimide (500 mg, 3.40 mmol) in acetone (25 mL) were added K2CO3 (1.88 g, 13.6 mmol), benzyltriethylammonium chloride (77.4 mg, 0.340 mmol), and 1,4-dibromobutane (2.94 g, 13.6 mmol). The reaction mixture was stirred at RT for 18 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n- hexane = 1 : 3 to 1 : 0) to obtain the title compound as a white solid (841 mg, 88%).

[0628] MS m / z: 283 [M+l]+. ’ H NMR (400 MHz, CDCI3) 5 7.84 (dd, J = 5.6, 2.9 Hz, 2H), 7.72 (dd, J = 5.6, 2.8 Hz, 2H), 3.73 (t, J = 6.9 Hz, 2H), 3.45 (t, J = 5.4 Hz, 2H), 1.96 - 1.83 (m, 4H).

[0629] [Preparative example C-21 (E)-2-(4-bromobut-2-en-l-yl)isoindoline- 1,3-dione (Int. C-2)

[0630] The title compound (457 mg, 60%) was synthesized following the procedure described in preparative example C-l using phthalimide potassium salt (500 mg, 2.70 mmol) and trans- 1 ,4-dibromo-2-butene (1.15 g, 5.40 mmol).

[0631] MS m / z: 281 [M+l]+.1H NMR (400 MHz, CDCI3) 5 7.91 - 7.82 (m, 2H), 7.77 - 7.70 (m, 2H), 6.00 - 5.90 (m, 1H), 5.87 - 5.79 (m, 1H), 4.31 (d, J = 7.0 Hz, 2H), 3.91 (d, J = 8.0 Hz, 2H).

[0632] [Preparative example C-31 tert-butyl (lH-pyrazol-4-yl)carbamate (Int. C-3)

[0633] Int. C-3

[0634] To a solution of 4-nitro-lH-pyrazole (6.97 g, 61.6 mmol) in MeOH (200 mL) were

[0635] -80-

[0636] FH13103096.12 Attorney Docket No.: FCH-03225 added Pd / C (3.5 g) and BOC2O (14.8 g, 67.8 mmol). After stirring at RT for 12 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as a white solid (9.17 g, 81%).

[0637] MS m / z: 184 [M+l]+. ’ H NMR (400 MHz, DMSO-rfo) 5 12.41 (s, 1H), 9.04 (s, 1H), 7.45 (s, 2H), 1.44 (s, 9H).

[0638] [Preparative example C-41 4-((tert-butvldimethvlsilvl)oxv)-lH-pyrazole (Int. C-4)

[0639] 1 nt. C-4

[0640] To a solution of lH-pyrazol-4-ol (500 mg, 5.95 mmol) in DMF (15 mL) were added imidazole (486 mg, 7.14 mmol) and tert-butyldimethylsilyl chloride (986 mg, 6.54 mmol) dissolved in DMF (5 mF) at RT. After stirring at RT for 18 h under nitrogen atmosphere, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mF), then extracted three times with chloroform (30 mF). The combined organic layers were washed with saturated NaHCCF aqueous solution, dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPFC (MeOH : DCM = 0 : 1 to 2 : 98) to obtain the title compound as a white solid (749 mg, 63%).

[0641] MS m / z: 199 [M+lf.'H NMR (400 MHz, CDCh) 57.21 (s, 2H), 0.96 (s, 9H), 0.17 (s, 6H).

[0642] The following intermediates Int.C-5 and Int.C-6 were synthesized in the same manner as described in preparative example C-4 using corresponding alcohols instead of IH-pyrazol- 4-ol.

[0643] [Preparative example C-51 3-bromo-5-((tert-butyldimethylsilyl)oxy)pyridine (Int. C-5) [Preparative example C-61 3-((tert-butyldimethylsilyl)oxy)pyrrolidine (Int. C-6)

[0644] -81-

[0645] FH13103096.12 Attorney Docket No.: LCH-03225

[0646] [Preparative example C-71 tert-butyl methyl(lH-pyrazol-4-yl)carbamate (Int. C-7)

[0647] (Step 1) tert-butyl (1 -benzyl- lH-pyrazol-4-yl)carbamate

[0648] To a solution of tert-butyl (lH-pyrazol-4-yl)carbamate (1.00 g, 5.46 mmol) in DMF (25 mL) were added K2CO3 (2.26 g, 16.4 mmol) and benzyl bromide (0.780 mL, 6.56 mmol) at 0 °C. The mixture was stirred at 60 °C for 6 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated. The resulting residue was purified by silica gel column chromatography (EA : n- hexane = 3 : 7) to obtain the title compound as a white solid (871 mg, 58%).

[0649] MS m / z: 274 [M+l]+.1H NMR (400 MHz, CDCh) 7.65 (s, 1H), 7.36 - 7.28 (m, 4H), 7.24 - 7.20 (m, 2H), 6.22 (s, 1H), 5.22 (s, 2H), 1.48 (s, 9H).

[0650] (Step 2) tert-butyl (1 -benzyl- lH-pyrazol-4-yl)(methyl)carbamate

[0651] To a solution of the compound prepared in (step 1) above (420 mg, 1.54 mmol) in DMF (15 mL) was added NaH (92 mg, 2.30 mmol, 60% in dispersion in mineral oil) at 0 °C. After stirring at 0 °C for 0.5 h, iodomethane (0.110 mL, 1.77 mmol) was added thereto. The reaction mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction was quenched by adding distilled water (20 mL), then extracted with EA (20 mL). The organic layer was dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 2 : 8) to obtain the title compound as a colorless oil (386 mg, 88%).

[0652] MS m / z: 288 [M+l]+.1H NMR (400 MHz, CDCh) 57.46 (s, 1H), 7.39 - 7.27 (m, 3H), 7.26 - 7.20 (m, 3H), 5.24 (s, 2H), 3.19 (s, 3H), 1.49 (s, 9H).

[0653] (Step 3) tert-butyl methyl(lH-pyrazol-4-yl)carbamate (Int. C-7)

[0654] To a solution of the compound prepared in (step 2) above (206 mg, 0.72 mmol) in MeOH (7 mL) was added Pd / C (200 mg). After stirring at 40 °C for 15 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to obtain the title compound as a colorless oil (141 mg, 100%), which was used for the next step without further purification.

[0655] MS m / z: 198 [M+l]+

[0656] -82-

[0657] FH13103096.12 Attorney Docket No.: LCH-03225

[0658] [Preparative example C-81 l-(lH-imidazol-2-yl)-N,N-dimethylmethanamine (Int. C-8)

[0659] (Step 1) 1 -trityl- IH-imidazole

[0660] To a solution of imidazole (1.00 g, 14.7 mmol) in DMF (15 mL) were added TEA (2.05 mL, 14.7 mmol) and trityl chloride (4.09 g, 14.7 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The remaining residue was filtered, washed with distilled water, then dried in vacuo to obtain the title compound as a white solid (2.76 g, 61%), which was used for the next step without further purification.

[0661] MS m / z: 311 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.46 (s, 1H), 7.37 - 7.31 (m, 9H), 7.18 - 7.11 (m, 6H), 7.07 (s, 1H), 6.82 (s, 1H).

[0662] (Step 2) 1 -trityl- 1 H-imidazole-2-carbaldehyde

[0663] The title compound (1.05 g, 48%) was synthesized following the procedure described in preparative example B-20 (step 2) using the compound (2.00 g, 6.44 mmol) obtained in (step 1) above.

[0664] MS m / z: 339 [M+l]+.1H NMR (400 MHz, CDCh) 59.23 (s, 1H), 7.36 - 7.27 (m, 10H), 7.16 - 7.07 (m, 6H), 7.02 (s, 1H).

[0665] (Step 3) N.N-dimethvl-l-(l-tritvl-lH-imidazol-2-yl)methanamine

[0666] To a solution of the compound prepared in (step 2) above (600 mg, 1.77 mmol) in MeOH (18 mL) was added 2 M dimethylamine solution in THF (1.07 mL, 2.13 mmol). After stirring at RT for 0.5 h, NaBH tCN (167 mg, 2.66 mmol) was added, then the reaction mixture was stirred at RT for 3 h under nitrogen atmosphere. On completion, the reaction was quenched by adding saturated NaHCOt aqueous solution (10 mL), diluted with distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the title compound as a white solid (287 mg, 44%).

[0667] MS m / z: 368 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.31 - 7.28 (m, 9H), 7.16 - 7.12 (m, 6H), 6.99 (d, J = 1.5 Hz, 1H), 6.72 (d, J = 1.5 Hz, 1H), 2.69 (s, 2H), 1.97 (s, 6H).

[0668] (Step 4) l-(lH-imidazol-2-yl)-N,N-dimethylmethanamine (Int. C-8)

[0669] To a solution of the compound prepared in (step 3) above (286 mg, 0.78 mmol) in DCM

[0670] (8 mL) was added 4 N HC1 aqueous solution in 1,4-dioxane (4 mL). The reaction mixture was -83-

[0671] FH13103096.12 Attorney Docket No.: LCH-03225 stirred at RT for 2 h under nitrogen atmosphere. On completion, the solvent was removed under reduced pressure. The resulting residue was filtered, washed with distilled water, then dried in vacuo to obtain the title compound as a light-yellow solid (153 mg), which was used for the next step without further purification.

[0672] 1H NMR (400 MHz, DMSO3) 5 7.67 (s, 2H), 4.52 (s, 2H), 2.79 (s, 6H).

[0673] [Preparative example C-91 3-(benzyloxy)-4-nitro-lH-pyrazole (Int. C-9) step 5 PBn

[0674] - NA

[0675] N°2

[0676] Int. C-9

[0677] (Step 1) 3-bromo-4-nitro-lH-pyrazole

[0678] To a solution of 3-bromo-lH-pyrazole (500 mg, 3.40 mmol) in sulfuric acid (2.5 mL) was added nitric acid (0.63 mL) at 0 °C. After stirring at 80 °C for 3 h, the reaction was quenched by adding saturated NaHCCL aqueous solution (30 mL) carefully, then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a yellow solid (343 mg, 53%), which was used for the next step without further purification.

[0679] MS m / z: 192 [M+l]+.1H NMR (400 MHz, DMSO-A) 5 14.33 (s, 1H), 8.98 (s, 1H).

[0680] (Step 2) 3-bromo-4-nitro- 1 -(tetrahydro-2H-pyran-2-yl)- 1 H-pyrazole

[0681] To a solution of the compound prepared in (step 1) above (343 mg, 1.79 mmol) in THF (4 mL) were added p-toluenesulfonic acid monohydrate (34.0 mg, 0.18 mmol) and 3,4-dihydro- 2H-pyran (0.33 mL, 3.57 mmol). The reaction mixture was stirred at 80 °C for 1 h under nitrogen atmosphere. After being cooled to RT, the solvent was removed under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n- hexane = 1 : 6) to obtain the title compound as a colorless oil (425 mg, 86%).

[0682] MS m / z: 277 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.39 (s, 1H), 5.40 - 5.33 (m, 1H), 4.13 - 4.03 (m, 1H), 3.77 - 3.69 (m, 1H), 2.22 - 2.12 (m, 1H), 2.04 - 1.91 (m, 2H), 1.75 - 1.65 (m, 3H).

[0683] (Step 3) 4-nitro- 1 -(tetrahydro-2H-pyran-2-yl)- lH-pyrazol-3-ol

[0684] A mixture of the compound prepared in (step 2) above (505 mg, 1.83 mmol) and KOH (821 mg, 14.6 mmol) dissolved in a mixed solvent of 1 ,2-dimethoxyethane / distilled water (15 mL, 2 : 3) was stirred at 100 °C for 15 h under nitrogen atmosphere. After being cooled to RT, the reaction was quenched by adding 1 N HC1 aqueous solution (100 mL), then extracted twice -84-

[0685] FH13103096.12 Attorney Docket No.: LCH-03225 with EA (100 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The title compound was obtained as a yellow solid (287 mg), which was used for the next step without further purification.

[0686] MS m / z: 214 [M+l]+

[0687] (Step 4) 3-(benzyloxy)-4-nitro- 1 -(tetrahydro-2H-pyran-2-yl)- IH-pyrazole

[0688] A mixture of the compound prepared in (step 3) above (287 mg, 1.35 mmol), K2CO3 (372 mg, 2.69 mmol), and benzyl bromide (0.16 mL, 1.35 mmol) dissolved in DMF (10 mL) was stirred at 80 °C for 1 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 1 : 4) to obtain the title compound as a yellow oil (72.1 mg, 13% for 2 steps).

[0689] MS m / z: 304 [M+l]+

[0690] (Step 5) 3-(benzyloxy)-4-nitro-lH-pyrazole (Int. C-9)

[0691] The compound prepared in (step 4) above (72.1 mg, 0.24 mmol) was dissolved in MeOH (2 mL), and cone. HC1 (0.1 mL) was added thereto. The reaction mixture was stirred at RT for 7 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with DCM (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 2 : 3) to obtain the title compound as a white solid (31.1 mg, 60%).

[0692] MS m / z: 220 [M+l]+.1H NMR (400 MHz, CDCI3) 59.77 (s, 1H), 8.15 (s, 1H), 7.48 (d, J = 7.5 Hz, 2H), 7.42 - 7.32 (m, 3H), 5.41 (s, 2H).

[0693] (Preparative example C-101 ethyl 4-((tert-butyldimethylsilyl)oxy)-lH-pyrazole-3- carboxylate (Int. C-10)

[0694] (Step 1) ethyl 2-diazo-3-oxobutanoate

[0695] To a solution of 4-acetamidobenzenesulfonyl azide (1.02 g, 4.23 mmol) in ACN (20 mL) were added ethyl acetoacetate (500 mg, 3.84 mmol) and TEA (1.6 mL, 11.5 mmol) at 0

[0696] -85-

[0697] FH13103096.12 Attorney Docket No.: LCH-03225

[0698] °C. The reaction mixture was stirred at RT for 18 h under nitrogen atmosphere. On completion, the reaction mixture was filtered, washed with ACN, and the filtrate was concentrated under reduced pressure. The remaining residue washed with Et2O, and the filtrate was concentrated under reduced pressure to obtain the title compound as a yellow oil (497 mg, 83%).

[0699] MS m / z: 157 [M+l]+.1H NMR (400 MHz, CDCh) 54.30 (q, J = 7.1 Hz, 2H), 2.48 (s, 3H), 1.33 (t, 7 = 7.1 Hz, 3H).

[0700] (Step 2) ethyl 3-((tert-butyldimethylsilyl)oxy)-2-diazobut-3-enoate (Int. C-10)

[0701] To a solution of the compound prepared in (step 1) above (496 mg, 3.18 mmol) in anhydrous DCM (10 mL) were added TEA (0.89 mL, 6.35 mmol) and tert-butyldimethylsilyl trifluoromethanesulfonate (0.730 mg, 3.18 mmol) at 0 °C. The reaction mixture was stirred at RT for 1 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with saturated NaHCCh aqueous solution (40 mL), then extracted twice with DCM (40 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as an orange oil (815 mg, 95%), which was used for the next step without further purification.

[0702] MS m / z: 271 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 5.00 (d, J = 2.0 Hz, 1H), 4.29 - 4.22 (m, 3H), 1.29 (t, J = 7.1 Hz, 3H), 0.91 (s, 9H), 0.22 (s, 6H).

[0703] (Step 3) ethyl 4-((tert-butyldimethylsilyl)oxy)-lH-pyrazole-3-carboxylate (Int. C-10)

[0704] The compound prepared in (step 2) above (815 mg, 3.01 mmol) was dissolved in benzotrifluoride (3.00 mL) and stirred at 100 °C for 15 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 3) to obtain the title compound as a white solid (145 mg, 18%).

[0705] MS m / z: 271 [M+l]+.1H NMR (400 MHz, CDCh) 5 10.28 (s, 1H), 7.23 (s, 1H), 4.37 (q, J = 7.1 Hz, 2H), 1.38 (t, J = 7.1 Hz, 3H), 1.00 (s, 9H), 0.20 (s, 6H).

[0706] [Preparative example C-111 tert-butyl 7-oxo-4,5,6,7-tetrahydro-lH-pyrrolo[2,3-c]pyridine- 1 -carboxylate (Int. C-ll)

[0707] (Step 1) ethyl 3-(2-((tert-butoxycarbonyl)amino)ethyl)-lH-pyrrole-2-carboxylate

[0708] To a solution of ethyl 2-isocyanoacetate (1.00 g, 8.84 mmol) in 1,4-dioxane (4.4 mL) were added silver carbonate (244 mg, 0.88 mmol) and tert-butyl but-3-yn-l-yl carbamate (2.24

[0709] -86-

[0710] FH13103096.12 Attorney Docket No.: LCH-03225 g, 13.3 mmol). The reaction mixture was stirred at 80 °C for 14 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA : n-hexane = 3 : 7) to obtain the title compound as a yellow oil (564 mg, 23%).

[0711] MS m / z: 283 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.94 (s, 1H), 6.85 (t, J = 2.8 Hz, 1H), 6.15 (t, J = 2.8 Hz, 1H), 4.70 (s, 1H), 4.36 - 4.28 (m, 2H), 3.42 - 3.30 (m, 2H), 2.98 (t, J = 6.8 Hz, 2H), 1.42 (s, 9H), 1.37 (t, J = 7.1 Hz, 3H).

[0712] (Step 2) 3-(2-((tert-butoxycarbonyl)amino)ethyl)-lH-pyrrole-2-carboxylic acid

[0713] To a solution of the compound prepared in (step 1) above (564 mg, 2.00 mmol) in a mixed solvent of MeOH / distilled water (10 mL, 1 : 1) was added LiOH monohydrate (168 mg, 4.00 mmol) at 0 °C. The reaction mixture was stirred at 50 °C for 15 h under nitrogen atmosphere. After being cooled to RT, the reaction was quenched by adding 1 N HC1 aqueous solution (30 mL), extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a light brown solid (466 mg, 92%), which was used for the next step without further purification.

[0714] MS m / z: 255 [M+l]+. ’ H NMR (400 MHz, DMSO-tfe) 5 11.38 (s, 1H), 6.92 - 6.76 (m, 2H), 6.01 (s, 1H), 3.13 - 3.02 (m, 2H), 2.87 - 2.75 (m, 2H), 1.36 (s, 9H).

[0715] (Step 3) l,4,5,6-tetrahydro-7H-pyrrolo[2,3-c]pyridin-7-one

[0716] To a solution of the compound prepared in (step 2) above (466 mg, 1.83 mmol) in DCM (18 mL) was added SOCh (160 pL, 2.20 mmol) at 0 °C. After stirring at RT for 3 h under nitrogen atmosphere, the reaction was quenched by adding saturated NaHCCL aqueous solution (100 mL), then extracted twice with DCM (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a light brown solid (194 mg, 78%), which was used for the next step without further purification.

[0717] MS m / z: 137 [M+l]+. ’ H NMR (400 MHz, DMSO-tfe) 5 11.50 (s, 1H), 7.08 (s, 1H), 6.84 (t, J = 2.7 Hz, 1H), 5.97 (t, J = 2.3 Hz, 1H), 3.33 - 3.29 (m, 2H), 2.66 - 2.60 (m, 2H).

[0718] (Step 4) tert-butyl 7-oxo-4, 5, 6, 7-tetrahydro-lH-pyrrolo[2,3-c]pyridine-l -carboxylate (Int. C-ll)

[0719] To a solution of the compound prepared in (step 3) above (194 mg, 1.42 mmol) in DCM (9.5 mL) were added BOC2O (327 pL, 1.42 mmol) and DMAP (8.70 mg, 0.07 mmol). The reaction mixture was stirred at RT for 16 h under nitrogen atmosphere. On completion, the

[0720] -87-

[0721] FH13103096.12 Attorney Docket No.: LCH-03225 reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (100% EA) to obtain the title compound as white solid (309 mg, 92%).

[0722] MS m / z: 237 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.33 (d, J = 3.1 Hz, 1H), 6.07 (d,

[0723] 7 = 3.1 Hz, 1H), 5.39 (s, 1H), 3.53 - 3.46 (m, 2H), 2.72 (t, J = 6.6 Hz, 2H), 1.61 (s, 9H). (Preparative example C-121 tert-butyl (R)-(5-oxopyrrolidin-3-yl)carbamate (Int. C-12)

[0724] (Step 1) (S)-5-oxopyrrolidin-3-yl methanesulfonate

[0725] To a solution of (4S)-4-hydroxypyrrolidin-2-one (100 mg, 0.99 mmol) in pyridine (5 mL) was added methanesulfonyl chloride (80.4 pL, 1.04 mmol) at 0 °C. The mixture was stirred at RT for 2 h under nitrogen atmosphere. The reaction was quenched by adding saturated NaHCCh aqueous solution (10 mL). The solvent was removed under reduced pressure. The remaining residue was filtered, washed with 50% MeOH in EA, then dried in vacuo to obtain the title compound as a brown solid (2.9 g, 97%), which was used for the next step without further purification.

[0726] MS m / z: 180 [M+l]+.1H NMR (400 MHz, DMSO-tfe) 5 7.84 (s, 1H), 5.35 - 5.29 (m, 1H), 3.65 (dd, 7 = 11.8, 5.3 Hz, 1H), 3.40 - 3.37 (m, 1H), 3.24 (s, 3H), 2.71 (dd, 7 = 17.7, 6.4

[0727] Hz, 1H), 2.30 - 2.24 (m, 1H).

[0728] (Step 2) (R)-4-azidopyrrolidin-2-one

[0729] A mixture of the compound prepared in (step 1) above (177 mg, 0.99 mmol) and NaNs (70.6 mg, 1.09 mmol) dissolved in DMF (3 mL) was stirred at 60 °C for 15 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with 25% i-PrOH in chloroform (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a brown solid (51.3 mg, 41%), which was used for the next step without further purification.

[0730] MS m / z: 127 [M+l]+.1H NMR (400 MHz, DMSO-tfe) 5 7.75 (s, 1H), 4.44 - 4.37 (m, 1H), 3.56 - 3.50 (m, 1H), 3.15 (dd, J = 15.1, 1.5 Hz, 1H), 2.58 (dd, J = 17.0, 7.1 Hz, 1H), 2.08 (dd, J = 17.0, 3.0 Hz, 1H).

[0731] (Step 3) tert-butyl (R)-(5-oxopyrrolidin-3-yl)carbamate (Int. C-12)

[0732] -88-

[0733] FH13103096.12 Attorney Docket No.: LCH-03225

[0734] The title compound (56.9 mg, 70%) was synthesized following the procedure described in preparative example C-3 using the compound (51.3 mg, 0.410 mmol) obtained in (step 2) above.

[0735] MS m / z: 201 [M+l]+.1H NMR (400 MHz, CDC13) 5 6.64 (s, 1H), 5.25 (s, 1H), 4.37 (s, 1H), 3.67 (dd, J = 10.4, 6.7 Hz, 1H), 3.22 (dd, J = 10.2, 4.1 Hz, 1H), 2.64 (dd, J = 17.3, 8.1 Hz, 1H), 2.19 (dd, J = 17.1, 4.9 Hz, 1H), 1.42 (s, 9H).

[0736] [Preparative example C-131 isothiazolidine 1,1 -dioxide (Int. C-13)

[0737] To a solution of 2-bromoethan-l -amine hydrobromide (200 mg, 0.98 mmol) in ACN (10 mL) were added methanesulfonyl chloride (79.3 pL, 1.02 mmol) and TEA (0.29 mL, 2.05 mmol) at 0 °C. The reaction mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion of the reaction, the resulting precipitate was filtered, washed with THF, dried in vacuo, then dissolved in n-hexane (1 mL). Diisopropylamine (41.3 pL, 0.29 mmol) and 1,10- phenantroline (1.76 mg, 0.01 mmol) were added thereto. After being cooled to -30 °C, 2 M n- BuLi solution in n-hexane (2.14 mL, 4.30 mmol) was added dropwise thereto. The reaction mixture was allowed to stir at the same temperature for 4 h under nitrogen atmosphere. The reaction was quenched by adding 1 N HC1 aqueous solution (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 8 : 2 to 1 : 1) to obtain the title compound as a brown oil (129 mg, 63%).

[0738] MS m / z: 122 [M+l]+.1H NMR (400 MHz, CDC13) 54.19 (s, 1H), 3.44 (q, J = 6.7 Hz, 2H), 3.10 (t, J = 7.4 Hz, 2H), 2.55 - 2.40 (m, 2H).

[0739] [Preparative example C-14] tert-butyl (2-oxopiperidin-3-yl)carbamate (Int. C-14)

[0740] (Step 1) 3-aminopiperidin-2-one

[0741] To a solution of 2,5 -diaminopentanoic acid hydrochloride (1.00 g, 5.93 mmol) was dissolved in distilled water (7 mL) was added NaHCCL (597 mg, 7.12 mmol). The reaction mixture was stirred at 120 °C for 1 h under nitrogen atmosphere. On completion, the solvent was removed under reduced pressure to obtain the title compound as a colorless oil (650 mg),

[0742] -89-

[0743] FH13103096.12 Attorney Docket No.: LCH-03225 which was used for the next step without further purification.

[0744] MS m / z: 115 [M+l]+

[0745] (Step 2) tert-butyl (2-oxopiperidin-3-yl)carbamate (Int. C-14)

[0746] The title compound (560 mg, 45%) was synthesized following the procedure described in preparative example C-ll (step 4) using the compound (650 mg, 5.69 mmol) obtained in (step 1) above.

[0747] MS m / z: 215 [M+l]+.1H NMR (400 MHz, CDCh) 5 5.79 (s, 1H), 5.42 (s, 1H), 3.34 - 3.36 (m, 2H), 2.57 - 2.44 (m, 1H), 1.97 - 1.84 (m, 2H), 1.62 - 1.54 (m, 2H), 1.45 (s, 9H).

[0748] (Preparative example C-151 8-oxa-2-azaspiro[4.5]decan-l-one (Int. C-15)

[0749] (Step 1) methyl 4-(cyanomethyl)tetrahydro-2H-pyran-4-carboxylate

[0750] To a solution of methyl tetrahydro-2H-pyran-4-carboxylate (300 mg, 2.08 mmol) in THF (6 mL) was added 2 M LDA solution in THF (1.14 mL, 2.29 mmol) at -78°C. The mixture was stirred at the same temperature for 40 min under nitrogen atmosphere. Bromoacetonitrile (0.17 mL, 2.5 mmol) and l,3-dimethyl-3,4,5,6-tetrahydro-2(lH)-pyrimidinone (0.12 mL, 1.0 mmol) were added thereto, then the mixture was stirred at RT for 15 h under nitrogen atmosphere. The reaction was quenched by adding 1 N HC1 aqueous solution (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 4) to obtain the title compound as a brown oil (146 mg, 38%).

[0751] MS m / z: 184 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 3.88 - 3.78 (m, 5H), 3.61 - 3.51 (m, 2H), 2.62 (s, 2H), 2.22 - 2.13 (m, 2H), 1.75 - 1.64 (m, 2H).

[0752] (Step 2) 8-oxa-2-azaspiro[4.5]decan-l-one (Int. C-15)

[0753] To a solution of the compound prepared in (step 1) above (146 mg, 0.80 mmol) in a mixed solvent of THF / distilled water (4.5 mL, 2 : 1) were added cobalt (II) chloride hexahydrate (94.6 mg, 0.40 mmol) and NaBH4 (150 mg, 3.98 mmol) at 0 °C. The reaction mixture was stirred at RT for 15 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure, and insoluble residue was filtered off. The filtrate was concentrated under reduced pressure, diluted with distilled water (50 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was

[0754] -90-

[0755] FH13103096.12 Attorney Docket No.: LCH-03225 purified by silica gel column chromatography (MeOH : EA = 1 : 9) to obtain the title compound as a white solid (31.4 mg, 25%).

[0756] MS m / z: 156 [M+l]+.1H NMR (400 MHz, CDCh) 5 5.48 (br s, 1H), 4.03 - 3.96 (m, 2H), 3.59 - 3.49 (m, 2H), 3.35 (td, J = 6.9, 0.9 Hz, 2H), 2.13 (t, J = 6.8 Hz, 2H), 2.07 - 1.95 (m, 2H), 1.43 - 1.34 (m, 2H).

[0757] [Preparative example C-161 8-oxa-2-azaspiro[4.5]decan-3-one (Int. C-16)

[0758] (Step 1) methyl 2-(tetrahydro-4H-pyran-4-ylidene)acetate

[0759] To a suspension of NaH (150 mg, 3.75 mmol, 60% in dispersion in mineral oil) in DMF (20 mL) was added trimethylphosphonoacetate (655 mg, 3.60 mmol) portionwise at 0 °C. After stirring at the same temperature for 50 min, tetrahydro-4H-pyran-4-one (300 mg, 3.00 mmol) was added thereto, then the reaction mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction was quenched by adding distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 1 : 10) to obtain the title compound as a colorless oil (366 mg, 78%).

[0760] MS m / z: 157 [M+l]+.1H NMR (400 MHz, CDCh) 55.69 (s, 1 H), 3.70 - 3.81 (m, 4 H), 3.70 (s, 3 H), 2.95 - 3.05 (m, 2 H), 2.30 - 2.37 (m, 2 H).

[0761] (Step 2) methyl 2-(4-(nitromethyl)tetrahydro-2H-pyran-4-yl)acetate

[0762] To a solution of the compound prepared in (step 1) above (150 mg, 0.96 mmol) in THF (5 mL) was added 1 M TBAF solution in THF (0.42 mL, 0.42 mmol). The mixture was stirred at RT for 0.5 h under nitrogen atmosphere, then nitromethane (117 mg, 1.92 mmol) was added thereto. The resulting mixture was stirred at 70 °C for 3 h under nitrogen atmosphere. After being cooled to RT, the reaction was quenched by adding 1 N HC1 aqueous solution (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 1 : 10 to 1 : 4) to obtain the title compound as a white solid (127 mg, 61%).

[0763] MS m / z: 218 [M+l]+

[0764] (Step 3) 8-oxa-2-azaspiro[4.5]decan-3-one (Int. C-16)

[0765] -91-

[0766] FH13103096.12 Attorney Docket No.: LCH-03225

[0767] To a solution of the compound prepared in (step 2) above (127 mg, 0.59 mmol) in MeOH (5 mL) was added Raney-Ni (34.0 mg). After stirring at 45°C for 18 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to obtain the tide compound as an off-white solid (68.0 mg), which was used for the next step without further purification.

[0768] MS m / z: 156 [M+l]+

[0769] [Preparative example C-17] tert-butyl 4-oxo-4,5,6,7-tetrahydro-lH-pyrrolo[3,2-c]pyridine- 1 -carboxylate (Int. C-17)

[0770] Int. C-17

[0771] (Step 1) tert-butyl 4-((2,2-diethoxyethyl)amino)-6-oxo-3,6-dihydropyridine-l(2H)- carboxylate

[0772] To a solution of tert-butyl 2,4-dioxopiperidine-l-carboxylate_(147 mg, 0.69 mmol) in toluene (2.3 mL) was added aminoacetaldehyde diethyl acetal (0.1 mL, 0.69 mmol). The reaction mixture was stirred at 70 °C for 3 h under nitrogen atmosphere. On completion, the solvent was removed under reduced pressure to obtain the title compound as a white solid (250 mg), which was used for the next step without further purification.

[0773] MS m / z: 329 [M+l]+

[0774] (Step 2) l,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridin-4-one

[0775] A mixture of the compound prepared in (step 1) above (250 mg) and TFA (1.5 mL) dissolved in DCM (5 mL) was stirred at RT for 1 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 9) to obtain the title compound as a colorless oil (52.0 mg, 55% for 2 steps).

[0776] MS m / z: 137 [M+l]+.1H NMR (400 MHz, CDCh) 5 6.73 - 6.69 (m, 1H), 6.62 - 6.56 (m, 1H), 3.63 (t, J = 6.3 Hz, 2H), 2.92 (t, . / = 7.1 Hz, 2H).

[0777] (Step 3) tert-butyl 4-oxo-4, 5, 6, 7-tetrahydro-lH-pyrrolo[3,2-c]pyridine-l -carboxylate (Int. C-17)

[0778] The title compound (45.7 mg, 51%) was synthesized following the procedure described in preparative example C-ll (step 4) using the compound (52.0 mg, 0.38 mmol) obtained in (step 2) above.

[0779] MS m / z: 237 [M+l]+.1H NMR (400 MHz, DMSO-rfe) 5 7.25 (s, 1H), 7.19 (d, J = 3.4

[0780] -92-

[0781] FH13103096.12 Attorney Docket No.: LCH-03225

[0782] Hz, 1H), 6.40 (d, J = 3.4 Hz, 1H), 3.43 - 3.38 (m, 2H), 3.06 (t, J = 7.0 Hz, 2H), 1.56 (s, 9H). [Preparative example C-181 tert-butyl (l-(3-aminopropyl)-lH-pyrazol-4-yl)carbamate (Int.

[0783] C-18)

[0784] (Step 1) tert-butyl (1 -(3-( 1 ,3-dioxoisoindolin-2-yl)propyl)- lH-pyrazol-4-yl)carbamate

[0785] A mixture of Int. C-3 (5.00 g, 27.3 mmol), N-(3-bromopropyl)phthalimide (8.78 g, 32.7 mmol), and CS2CO3 (9.78 mg, 30.0 mmol) in DMF (25 mL) was stirred at 60 °C for 4 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as a light brown solid (9.35 g, 92%).

[0786] MS m / z: 371 [M+l]+.1H NMR (400 MHz, CDCI3) 5 7.83 (d, J = 4 Hz, 2H), 7.73-7.72 (m, 3H), 7.31 (s, 1H), 6.23 (s, 1H), 4.13 - 4.08 (m, 2H), 3.73 (t, J = 6.8 Hz, 2H), 2.31 - 2.21 (m, 2H), 1.49 (s, 9H).

[0787] (Step 2) tert-butyl (l-(3-aminopropyl)-lH-pyrazol-4-yl)carbamate (Int. C-18)

[0788] A mixture of the compound prepared in (step 1) above (2.10 g, 5.67 mmol) and hydrazine monohydrate (138 pL, 28.3 mmol) dissolved in EtOH (3 mL) was stirred at 50 °C for 2 h under nitrogen atmosphere. After being cooled to RT, the reaction mixture was filtered to remove the precipitate. The filtrate was concentrated under reduced pressure to obtain the title compound as a white solid (1.40 g, 99%), which was used for the next step without further purification.

[0789] MS m / z: 241 [M+l]+.1H NMR (400 MHz, CDCI3) 57.66 (s, 1H), 7.29 (s, 1H), 6.26 (s, 1H), 4.14 (t, J = 6.8 Hz, 2H), 2.73 - 2.65 (m, 2H), 1.98-1.93 (m, 2H), 1.49 (s, 9H).

[0790] [Preparative example C-191 3-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)propan-l-amine (Int. C-19)

[0791] (Step 1) 2-(3-(7H-pvrrolo[2,3-d1pvrimidin-7-vl)propvl)isoindoline-1.3-dione

[0792] A mixture of N-(3-bromopropyl)phthalimide (135 mg, 0.50 mmol), 7H-pyrrolo[2,3-d] pyrimidine (50 mg, 0.42 mmol), and K2CO3 (87.0 mg, 0.63 mmol) dissolved in DMF (2 mL) was stirred at 80 °C for 14 h under nitrogen atmosphere. On completion, the reaction mixture -93-

[0793] FH13103096.12 Attorney Docket No.: LCH-03225 was concentrated under reduced pressure. The residue was diluted with distilled water (30 ruL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 7 : 3) to obtain the title compound as a pink solid (88.0 mg, 68%).

[0794] MS m / z: 307 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.93 (s, 1H), 8.82 (s, 1H), 7.87 - 7.82 (m, 2H), 7.77 - 7.70 (m, 2H), 7.38 (d, J = 3.6 Hz, 1H), 6.55 (d, J = 3.5 Hz, 1H), 4.35 (t, J = 6.9 Hz, 2H), 3.75 (t, J = 6.8 Hz, 2H), 2.37 - 2.25 (m, 2H).

[0795] (Step 2) 3-(7H-pyrrolo[2,3-d]pyrimidin-7-yl)propan-l-amine (Int. C-19)

[0796] The title compound (55.2 mg, 63%) was synthesized following the procedure described in preparative example C-18 (step 2) using the compound (88.0 g, 0.29 mmol) obtained in (step 1) above.

[0797] MS m / z: 177 [M+l]+.1H NMR (400 MHz, CDCh) 58.95 (s, 1H), 8.87 (s, 1H), 7.24 (d, 7 = 3.5 Hz, 1H), 6.56 (d, 7 = 3.5 Hz, 1H), 4.39 (t, 7 = 6.8 Hz, 2H), 2.66 (t, 7 = 6.6 Hz, 2H), 2.06 - 1.95 (m, 2H).

[0798] The following intermediates were synthesized in the same manner as described in preparative example C-18 (Int. C-20 ~ Int. C-24) or preparative example C-19 (Int. C-25 ~ Int.C-28) using the corresponding phthalimides and amines.

[0799] [Preparative example C-201 tert-butyl (l-(3-aminopropyl)-2-oxopyrrolidin-3-yl) carbamate (Int. C-20)

[0800] [Preparative example C-21] l-(3-aminopropyl)-lH-imidazol-2-amine (Int. C-21)

[0801] [Preparative example C-221 3-(2-((dimethylamino)methyl)-lH-imidazol-l-yl) propan-1- amine (Int. C-22)

[0802] [Preparative example C-231 l-(3-aminopropyl)pyridin-2(lH)-one (Int. C-23)

[0803] [Preparative example C-241 l-(4-aminobutyl)-lH-imidazol-2-amine (Int. C-24)

[0804] [Preparative example C-251 tert-butyl (l-(3-aminopropyl)pyrrolidin-3-yl)carbamate (Int. C- 25)

[0805] [Preparative example C-261 tert-butyl (E)-( 1 -(4-aminobut-2-en- 1 -yl)- 1 H-pyrazol-4- yl)carbamate (Int. C-26)

[0806] [Preparative example C-271 l-(3-aminopropyl)-3-nitropyridin-2(lH)-one (Int. C-27) [Preparative example C-281 3-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin- 1 -yl)propan- 1 - amine (Int. C-28)

[0807] -94-

[0808] FH13103096.12 Attorney Docket No.: LCH-03225

[0809] FH13103096.12 Attorney Docket No.: LCH-03225

[0810] [Preparative example C-291 3-amino-l-(3-aminopropyl)pyridin-2(lH)-one (Int. C-29)

[0811] Int. C-29

[0812] The title compound (25.0 mg, 98%) was synthesized following the procedure described in preparative example A-12 (step 1) using Int. C-27 (30.0 mg, 0.15 mmol).

[0813] MS m / z: 168 [M+l]+

[0814] [Preparative example C-301 l-(3-aminopropyl)pyrrolidin-3-ol (Int. C-30) lnt.C-30

[0815] To a solution of Int. C-28 (67.1 mg, 0.26 mmol) in DCM (2 mL) was added 4 N HC1 solution in 1,4-dioxane (1 mL). The reaction mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the solvent was removed under reduced pressure. The title compound was obtained as a white solid (75.4 mg), which was used for the next step without further purification.

[0816] MS m / z: 145 [M+l]+

[0817] [Preparative example C-311 1 -(3-aminopropyl)-2-oxo- 1 ,2-dihydropyridine-3-carboxamide (Int. C-31)

[0818] (Step 1) l-(3-(l,3-dioxoisoindolin-2-yl)propyl)-2-oxo-l,2-dihydropyridine-3- carbonitrile

[0819] The title compound (200 mg, 78%) was synthesized following the procedure described in preparative example C-19 (step 1) using hydroxypyridine-3-carbonitrile (100 mg, 0.83 mmol) instead of 7H-pyrrolo[2,3-d]pyrimidine.

[0820] MS m / z: 308 [M+l]+

[0821] (Step 2) l-(3-(l,3-dioxoisoindolin-2-yl)propyl)-2-oxo-l,2-dihydropyridine-3- carboxamide

[0822] A solution of the compound prepared in (step 1) above (60.0 mg, 0.20 mmol) in sulfuric acid (0.72 mL) was stirred at RT for 16 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (30 mL), then extracted with EA (30 mL). The -96-

[0823] FH13103096.12 Attorney Docket No.: LCH-03225 organic layer was dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The title compound was obtained as a white solid (55.0 mg, 86%), which was used for the next step without further purification.

[0824] MS m / z: 326 [M+l]+

[0825] (Step 3) l-(3-aminopropyl)-2-oxo-l,2-dihydropyridine-3-carboxamide (Int. C-31)

[0826] The title compound (27.0 mg, 80%) was synthesized following the procedure described in preparative example C-18 (step 2) using the compound (55.8 mg, 0.17 mmol) obtained in (step 2) above.

[0827] MS m / z: 196 [M+l]+

[0828] [Preparative example C-32] 4-amino-l-(3-aminopropyl)-lH-pyrazol-3-ol (Int. C-32)

[0829] Int. C-32

[0830] ( Step 1 ) 2-( 3-(3-(benzyloxy)-4-nitro- 1 H-pyrazol- 1 -vDpropvDisoindoline- 1 ,3-dione

[0831] The title compound (62.7 mg, 88%) was synthesized following the procedure described in preparative example C-19 (step 1) using N-(3-bromopropyl)phthalimide (56.5 mg, 0.21 mmol) and Int. C-9 (38.5 mg, 0.180 mmol).

[0832] MS m / z: 407 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 8.15 (s, 1H), 7.85 - 7.81 (m, 2H), 7.74 - 7.69 (m, 2H), 7.49 - 7.43 (m, 2H), 7.40 - 7.34 (m, 2H), 7.34 - 7.29 (m, 1H), 5.32 (s, 2H), 4.02 (t, J = 6.5 Hz, 2H), 3.73 (t, J = 6.3 Hz, 2H), 2.32 - 2.24 (m, 2H).

[0833] (Step 2) 2-(3-(4-amino-3-hydroxy- IH-pyrazol- 1 -yl)propyl)isoindoline- 1 ,3-dione

[0834] The title compound (35.6 mg, 81%) was synthesized following the procedure described in preparative example C-29 using the compound (38.5 mg, 0.18 mmol) obtained in (step 1) above.

[0835] MS m / z: 287 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.85 - 7.81 (m, 2H), 7.73 - 7.70 (m, 2H), 7.01 (s, 1H), 3.85 (t, J = 6.7 Hz, 2H), 3.70 (t, J = 6.6 Hz, 2H), 2.18 - 2.12 (m, 2H).

[0836] (Step 3) 4-amino-l-(3-aminopropvl)-lH-pyrazol-3-ol (Int. C-32)

[0837] The title compound (23.2 mg) was synthesized following the procedure described in preparative example C-18 (step 2) using the compound (35.6 mg, 0.12 mmol) obtained in (step 2) above.

[0838] MS m / z: 157 [M+l]+

[0839] -97-

[0840] FH13103096.12 Attorney Docket No.: LCH-03225

[0841] [Preparative example C-331 tert-butyl ( 1 -(3-amino-2-hydroxypropyl)- 1 H-pyrazol-4- yl)carbamate (Int. C-33)

[0842] (Step 1) 2-(oxiran-2-ylmethyl)isoindoline- 1 ,3-dione

[0843] The title compound (1.53 g, 64%) was synthesized following the procedure described in preparative example C-l using phthalimide potassium salt (1.73 g, 11.7 mmol) and 2- (bromomethyl)oxirane.

[0844] MS m / z: 204 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.90 - 7.82 (m, 2H), 7.77 - 7.60 (m, 2H), 3.99 - 3.85 (m, 1H), 3.83 - 3.70 (m, 1H), 3.30 - 3.27 (m, 1H), 2.81 (t, J = 4.4 Hz, 1H), 2.75 - 2.69 (m, 1H).

[0845] (Step 2) tert-butyl (l-(3-(l,3-dioxoisoindolin-2-yl)-2-hydroxypropyl)-lH-pyrazol-4- yl)carbamate

[0846] A mixture of the compound prepared in (Step 1) above (381 mg, 1.87 mmol), Int. C-2 (286 mg, 1.56 mmol), and CS2CO3 (560 mg, 1.72 mmol) dissolved in ACN (5 mL) was stirred at 90 °C for 4 h. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 1 : 1) to obtain the title compound as a light yellow solid (145 mg, 24%).

[0847] MS m / z: 387 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.94 - 7.80 (m, 2H), 7.78 - 7.60 (m, 3H), 7.37 (s, 1H), 4.30 (s, 1H), 4.24 (d, J = 13.8 Hz, 1H), 4.09 - 4.15 (m, 2H), 3.94 (s, 1H), 3.87 - 3.71 (m, 2H), 1.49 (s, 9H).

[0848] (Step 3) tert-butyl (l-(3-amino-2-hydroxypropyl)-lH-pyrazol-4-yl)carbamate (Int. C- 33)

[0849] The title compound (126 mg) was synthesized following the procedure described in preparative example C-l 8 (step 2) using the compound (172 mg, 0.45 mmol) obtained in (step 2).

[0850] MS m / z: 257 [M+l]+.1H NMR (400 MHz, CDCh) 57.70 (s, 1H), 7.36 (s, 1H), 6.23 (s, 1H), 5.30 (s, 1H), 4.08 - 4.13 (m, 2H), 3.96 - 3.87 (m, 1H), 2.79 - 2.82 (m, 1H), 2.65 - 2.67 (m, 1H), 1.50 (s, 9H).

[0851] [Preparative example C-341 (E)-3-(4-aminobut-2-en-l-yl)oxazolidin-2-one (Int. C-34)

[0852] FH13103096.12 Attorney Docket No.: LCH-03225

[0853] (Step 1) (E)-3-(4-bromobut-2-en- l-yl)oxazolidin-2-one

[0854] To a solution of 2-oxazolidinone (200 mg, 2.30 mmol) in THF (11 mL) was added 1 M NaHMDS solution in THF (2.3 mL) at -70 °C. The mixture was stirred at the same temperature for 20 min under nitrogen atmosphere. Trans- l,4-dibromo-2-butene (737 mg, 3.45 mmol) dissolved in THF (2 mL) was added thereto, then the mixture was stirred at RT for 19 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (10 mL), then extracted twice with EA (10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as a colorless oil (183 mg, 36%).

[0855] 1H NMR (400 MHz, CDCh) 5 5.97 - 5.85 (m, 1H), 5.79 - 5.69 (m, 1H), 4.39 - 4.29 (m, 2H), 3.99 - 3.92 (m, 2H), 3.90 (dt, . / = 6.3, 1.1 Hz, 2H), 3.57 - 3.49 (m, 2H).

[0856] (Step 2) (E)-2-(4-(2-oxooxazolidin-3-yl)but-2-en-l-yl)isoindoline- 1,3-dione

[0857] The title compound (158 mg, 66%) was synthesized following the procedure described in preparative example C-l using the compound (183 mg, 0.83 mmol) obtained in (step 1) above.

[0858] 1H NMR (400 MHz, CDCh) 5 7.91 - 7.81 (m, 2H), 7.78 - 7.70 (m, 2H), 5.81 - 5.70 (m, 1H), 5.69 - 5.60 (m, 1H), 4.35 - 4.25 (m, 4H), 3.90 - 3.79 (m, 2H), 3.52 - 3.42 (m, 2H).

[0859] (Step 3) (E)-3-(4-aminobut-2-en-l-yl)oxazolidin-2-one (Int. C-34)

[0860] The title compound (85.4 mg, 98%) was synthesized following the procedure described in preparative example C-l 8 (step 2) using the compound (158 mg, 0.55 mmol) obtained in (step 2) above.

[0861] MS m / z: 157 [M+l]+.1H NMR (400 MHz, CDCh) 5 5.85 - 5.77 (m, 1H), 5.61 - 5.52 (m, 1H), 4.35 - 4.30 (m, 2H), 3.89 - 3.83 (m, 2H), 3.55 - 3.49 (m, 2H), 3.36 - 3.31 (m, 2H), 1.68 - 1.54 (m, 2H).

[0862] [Preparative example C-351 l-(4-aminobutyl)pyrrolidin-2-one (Int. C-35)

[0863] (Step 1) 1 -(4-bromobutyl)pyrrolidin-2-one

[0864] To a solution of 2-pyrrolidinone (100 mg, 1.18 mmol) in DMF (12 mL) was added NaH

[0865] FH13103096.12 Attorney Docket No.: LCH-03225

[0866] (94.0 mg, 2.35 mmol, 60% in dispersion in mineral oil) at 0 °C. After stirring at the same temperature for 10 min, 1 ,4-dibromobutane (380 mg, 1.76 mmol) was added to the reaction mixture. After stirring at RT for 18 h under nitrogen atmosphere, the reaction was quenched by adding distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were washed with distilled water three times, dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-Hexane= 1 : 10) to obtain the title compound as a colorless oil (193.0 mg, 74.6%).

[0867] MS m / z: 220 [M]+

[0868] (Step 2) 2-(4-(2-oxopyrrolidin- 1 -yl)butyl)isoindoline- 1 ,3-dione

[0869] The title compound (60.3 mg, 78%) was synthesized following the procedure described in preparative example C-l using the compound (50.0 mg, 0.27 mmol) obtained in (step 1) above.

[0870] MS m / z: 287 [M+l]+

[0871] (Step 3) l-(4-aminobutyl)pyrrolidin-2-one (Int. C-35)

[0872] The title compound (32.9 mg, 99%) was synthesized following the procedure described in preparative example C-l 8 (step 2) using the compound (60.3 mg, 0.21 mmol) obtained in (step 2) above.

[0873] MS m / z: 157 [M+l]+

[0874] (Preparative example C-361 tert-butyl (l-(3-amino-2,2-dimethylpropyl)-lH-pyrazol-4- yl)carbamate (Int. C-36)

[0875] (Step 1) 2-(3-hydroxy-2,2-dimethylpropyl)isoindoline- 1 ,3-dione

[0876] A mixture of potassium l,3-dioxoisoindolin-2-ide (1.11 g, 5.99 mmol) and 3-bromo- 2,2-dimethylpropan-l-ol (1.00 g, 5.99 mmol) dissolved in DMF (5 mL) was stirred at 135 °C for 16 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (0.2% formic acid in ACN : distilled water = 5 : 95 to 100 : 0) to obtain the tide compound as a white solid (725 mg, 52%).

[0877] MS m / z: 234 [M+l]+. ’ H NMR (400 MHz, CDC13) 5 7.87 (dd, J = 3.1, 5.3 Hz, 2H), 7.78 - 7.72 (m, 2H), 3.60 (s, 2H), 3.58 (br s, 1H), 3.23 - 3.15 (m, 2H), 0.97 (s, 6H).

[0878] -100-

[0879] FH13103096.12 Attorney Docket No.: LCH-03225

[0880] (Step 2) 2-(2,2-dimethyl-3-(4-nitro- IH-pyrazol- 1 -yl)propyl)isoindoline- 1 ,3-dione

[0881] To a solution of the compound prepared in (step 1) above (100 mg, 0.43 mmol) and 4- nitro-lH-pyrazole (58.0 mg, 0.51 mmol) in THF (4 mL) were added PPht (168 mg, 0.64 mmol) and DIAD (0.13 mL, 0.64 mmol) sequentially at 0 °C. The mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 3) to obtain the title compound as a colorless oil (118 mg, 84%).

[0882] MS m / z: 329 [M+l]+

[0883] (Step 3) tert-butyl (l-(3-(l,3-dioxoisoindolin-2-yl)-2,2-dimethylpropyl)-lH-pyrazol-4- yl)carbamate

[0884] The tide compound (13.6 mg, 8%) was synthesized following the procedure described in preparative example C-3 using the compound (118 mg, 0.36 mmol) obtained in (step 2) above.

[0885] MS m / z: 399 [M+l]+

[0886] (Step 4) tert-butyl (l-(3-amino-2,2-dimethylpropyl)-lH-pyrazol-4-yl)carbamate (Int. C-36)

[0887] The title compound (9.70 mg, 99%) was synthesized following the procedure described in preparative example C-18 (step 2) using the compound (13.6 mg, 0.03 mmol) obtained in (step 3) above.

[0888] MS m / z: 269 [M+l]+

[0889] (Preparative example C-371 tert-butyl (l-((3-aminocyclobutyl)methyl)-lH-pyrazol-4- yl)carbamate (Int. C-37)

[0890] Int. C-37

[0891] The title compound was synthesized in the same manner as described in preparative example C-36 using (3-bromocyclobutyl)methanol instead of 3-bromo-2,2-dimethylpropan-l- ol.

[0892] MS m / z: 267 [M+l]+

[0893] (Preparative example C-381 tert-butyl (l-((3-(aminomethyl)oxetan-3-yl)methyl)-lH- pyrazol-4-yl)carbamate (Int. C-38)

[0894] -101-

[0895] FH13103096.12 Attorney Docket No.: LCH-03225

[0896] Int. C-38

[0897] The title compound was synthesized in the same manner as described in preparative example C-36 using (3-(bromomethyl)oxetan-3-yl)methanol instead of 3-bromo-2,2- dimethylpropan- l-ol.

[0898] MS m / z: 283 [M+l]+

[0899] [Preparative example C-391 l-(3-aminopropyl)-lH-l,2,4-triazol-3-amine (C-39)

[0900] Int. C-39

[0901] (Step 1) 2-(3-(3-nitro- 1H- 1 ,2, 4-triazol- 1 -yl)propyl)isoindoline- 1 ,3-dione

[0902] The title compound (191 mg, 72%) was synthesized following the procedure described in preparative example C-19 (step 1) using N-(3-bromopropyl)phthalimide (259 mg, 0.96 mmol).

[0903] MS m / z: 302 [M+l]+.1H NMR (400 MHz, DMSO-&) 5 8.80 (s, 1H), 7.90 - 7.77 (m, 4H), 4.38 (t, J = 7.0 Hz, 2H), 3.63 (t, J = 6.4 Hz, 2H), 2.22 (p, J = 6.6 Hz, 2H).

[0904] (Step 2) 2-(3-(3-amino-lH-1.2.4-triazol-l-vl)propyl)isoindoline-1.3-dione

[0905] The title compound (167 mg, 97%) was synthesized following the procedure described in preparative example C-29 using the compound (191 mg, 0.63 mmol) obtained in (step 1) above.

[0906] MS m / z: 272 [M+l]+.1H NMR (400 MHz, DMSO-tfe) 5 7.94 - 7.78 (m, 5H), 5.21 (s, 2H), 3.95 (t, J = 6.6 Hz, 2H), 3.55 (t, J = 6.6 Hz, 2H), 2.04 (p, J = 6.6 Hz, 2H).

[0907] (Step 3) l-(3-aminopropyl)-lH-l,2,4-triazol-3-amine (Int. C-39)

[0908] The title compound (94.5 mg) was synthesized following the procedure described in preparative example C-18 (step 2) using the compound (167 mg, 0.62 mmol) obtained in (step 2) above.

[0909] MS m / z: 142 [M+l]+

[0910] [Preparative example C-40] tert-butyl 4-(3-hydroxypropyl)-3-oxopiperazine- 1 -carboxylate (Int. C-40)

[0911] -102-

[0912] FH13103096.12 Attorney Docket No.: LCH-03225

[0913] (Step 1) tert-butyl 3-oxopiperazine-l-carboxylate

[0914] The title compound (401 mg, 80%) was synthesized following the procedure described in preparative example C-ll (step 4) using 2-piperazinone (200 mg, 2.00 mmol).

[0915] MS m / z: 201 [M+l]+.1H NMR (400 MHz, CDCh) 54.10 (s, 2H), 3.64 (t, J = 5.2 Hz,

[0916] 2H), 3.35 - 3.44 (m, 2H), 1.48 (s, 9H).

[0917] (Step 2) tert-butyl 4-(3-((tert-butyldimethylsilyl)oxy)propyl)-3-oxopiperazine- 1 - carboxylate

[0918] The title compound (123 mg, 53%) was synthesized following the procedure described in preparative example C-35 (step 1) using the compound (113 mg, 0.56 mmol) obtained in (step 1) above.

[0919] MS m / z: 373 [M+l]+

[0920] (Step 3) tert-butyl 4-(3-hydroxypropyl)-3-oxopiperazine-l -carboxylate (Int. C-40)

[0921] To a solution of the compound prepared in (step 2) above (123 mg, 0.33 mmol) in a mixed solvent of THF / distilled water (3 mL, 1 : 1) was added acetic acid (4.0 mL, 69.3 mmol), then the reaction mixture was stirred at 40 °C for 2 h under nitrogen atmosphere. After being cooled to RT, the reaction mixture was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (MeOH : DCM = 1 : 19) to obtain the tide compound as a colorless oil (85.0 mg, 99%).

[0922] MS m / z: 259 [M+l]+.1H NMR (400 MHz, CDCh) 54.11 (s, 2H), 3.65 (t, J = 5.4 Hz, 2H), 3.56 (dt, J = 11.1, 5.4 Hz, 5H), 3.35 (t, J = 5.4 Hz, 2H), 1.75 (t, J = 6.1 Hz, 2H), 1.48 (s,

[0923] 9H).

[0924] (Preparative example C-411 tert-butyl ( 1 -(2 ,2-difluoro-3-hydroxypropyl)- 1 H-pyrazol-4- yl)carbamate (Int. C-41)

[0925] (Step 1) 2,2-difluoropropane-l,3-diol

[0926] To a solution of dimethyl 2,2-difluoropropanedioate (1.00 g, 5.95 mmol) in THF (12

[0927] -103-

[0928] FH13103096.12 Attorney Docket No.: LCH-03225 mL) was added 1 M LiAlE solution in THF (11.9 mL, 11.9 mmol) at 0 °C, then the resulting mixture was stirred for 3 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (100 mL), then 15% NaOH aqueous solution (100 mL) added thereto slowly. The resulting slurry was filtered through a Celite pad, and the filtrate was extracted twice with EA (200 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n- hexane = 2 : 1 to 1 : 0) to obtain the title compound as a white solid (540 mg, 81%).

[0929] MS m / z: 113 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 3.97 - 3.85 (m, 4H), 1.91 (t, J = 7.1 Hz, 2H).

[0930] (Step 2) 2,2-difluoro-3-(trityloxy)propan- 1 -ol

[0931] To a solution of the compound prepared in (step 1) above (150 mg, 1.34 mmol) in a mixed solvent of DCM / THF (5 mL, 4 : 1), were added DIPEA (230 pL, 1.34 mmol), trityl chloride (373 mg, 1.34 mmol), and DMAP (16.4 mg, 0.13 mmol). The reaction mixture was stirred at 40 °C for 7 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 5) to obtain the title compound as a white solid (156 mg, 33%).

[0932] MS m / z: 355 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.47 - 7.40 (m, 6H), 7.34 - 7.28 (m, 6H), 7.27 - 7.22 (m, 3H), 3.91 (t, 7 = 13.1 Hz, 2H), 3.42 (t, J = 12.2 Hz, 2H), 1.95 (s, 1H).

[0933] (Step 3) l-(2,2-difluoro-3-(trityloxy)propyl)-4-nitro-lH-pyrazole

[0934] The title compound (121 mg, 95%) was synthesized following the procedure described in preparative example C-36 (step 2) using the compound (100 mg, 0.03 mmol) obtained in (step 2) above.

[0935] MS m / z: 450 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.11 (s, 1H), 7.98 (s, 1H), 7.47 - 7.39 (m, 6H), 7.36 - 7.30 (m, 6H), 7.29 - 7.24 (m, 3H), 4.68 (t, J = 13.0 Hz, 2H), 3.30 (t, J = 11.7 Hz, 2H).

[0936] (Step 4) 2,2-difluoro-3-(4-nitro-lH-pyrazol-l-vl)propan-l-ol

[0937] A mixture of the compound prepared in (step 3) above (163 mg, 0.36 mmol) and TFA (1.5 mL) dissolved in DCM (3 mL) was stirred at RT for 1 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (15 mL), then extracted twice with EA (15 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-

[0938] -104-

[0939] FH13103096.12 Attorney Docket No.: LCH-03225 hexane = 2 : 3) to obtain the title compound as a white solid (54.5 mg, 73%).

[0940] MS m / z: 208 [M+l]+1H NMR (400 MHz, CDCh) 58.29 (s, 1H), 8.14 (s, 1H), 4.66 (t, J = 12.4 Hz, 2H), 3.75 (t, J = 12.3 Hz, 2H), 3.12 (s, 1H).

[0941] (Step 5) tert-butyl (l-(2,2-difluoro-3-hydroxypropyl)-lH-pyrazol-4-yl)carbamate (Int.

[0942] C-41)

[0943] The title compound (67.6 mg, 93%) was synthesized following the procedure described in preparative example C-3 using the compound (54.5 mg, 0.26 mmol) obtained in (step 4).

[0944] MS m / z: 278 [M+l]+. ’ H NMR (400 MHz, CDCh) 57.72 (s, 1H), 7.41 (s, 1H), 6.51 (s, 1H), 4.49 (t, J = 12.1 Hz, 2H), 4.04 (s, 1H), 3.57 (t, J = 12.6 Hz, 2H), 1.49 (s, 9H).

[0945] [Preparative example C-42] tert-butyl 2-(3-hydroxypropyl)-3-oxotetrahydropyridazine- l(2H)-carboxylate (Int. C-42)

[0946] (Step 1) 3-(benzyloxy)propan-l-ol

[0947] To a solution of propane- 1 ,3-diol (2.00 g, 26.3 mmol) was dissolved in DMF (30 mL) was added NaH (1.16 g, 28.9 mmol, 60% in dispersion in mineral oil) at 0 °C. After stirring at the same temperature for 10 min, benzyl bromide (3.13 mL, 26.3 mmol) was added thereto, then the resulting mixture was stirred at RT for 18 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (100 mL) at 0 °C, then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was passed through a short column of silica gel (EA : n-hexane = 1 : 1), and the resulting colorless oil (3.33 g) was used for the next step without further purification.

[0948] MS m / z: 167 [M+l]+.1H NMR (400 MHz, CDCh) 57.39 - 7.27 (m, 5H), 4.53 (s, 2H),

[0949] 3.82 - 3.76 (m, 2H), 3.67 (t, J = 5.8 Hz, 2H), 2.28 - 2.22 (m, 1H), 1.91 - 1.83 (m, 2H).

[0950] (Step 2) 3-(benzyloxy)propanal

[0951] To a solution of the compound prepared in (step 1) above (3.33 g) in DCM (30 mL) was added Dess-Martin periodinane (7.48 g, 17.6 mmol) carefully at 0 °C with stirring. After stirring at RT for 1 h under nitrogen atmosphere, the reaction was quenched by adding 1 N HC1 aqueous solution (100 mL), then extracted twice with DCM (100 mL). The combined organic

[0952] -105-

[0953] FH13103096.12 Attorney Docket No.: LCH-03225 layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 4) to obtain the title compound as a colorless liquid (2.26 g, 52% for 2 steps).

[0954] MS m / z: 165 [M+l]+.1H NMR (400 MHz, CDCh) 59.80 (s, 1H), 7.37 - 7.27 (m, 5H), 4.54 (s, 2H), 3.82 (t, J = 6.1 Hz, 2H), 2.74 - 2.67 (m, 2H).

[0955] (Step 3) tert-butyl 2-(3-(benzyloxy)propyl)hydrazine-l -carboxylate

[0956] To a solution of the compound prepared in (step 2) above (2.26 g, 13.8 mmol) in THF (25 mL) was added tert-butyl carbazate (1.82 g, 13.8 mmol). The reaction mixture was stirred at RT for 15 h under nitrogen atmosphere. On completion, the mixture was concentrated under reduced pressure. The remaining residue was dissolved in toluene (30 mL), and 1 M DIBAL- H solution in cyclohexane (33 mL, 33.0 mmol) was added thereto at 0 °C. After stirring at RT for 3 h under nitrogen atmosphere, the reaction was quenched by adding 1 N NaOH aqueous solution (30 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as a colorless oil (1.50 g, 39%).

[0957] MS m / z: 281 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.36 - 7.30 (m, 5H), 6.01 (br s, 1H), 4.51 (s, 2H), 4.03 (br s, 1H), 3.56 (t, J = 6.3 Hz, 2H), 2.96 (t, J = 6.9 Hz, 2H), 1.82 - 1.74 (m, 2H), 1.46 (s, 9H).

[0958] (Step 4) tert-butyl 2-(3-(benzyloxy)propyl)-3-oxotetrahydropyridazine- 1 (2H)- carboxylate

[0959] To a solution of the compound prepared in (step 3) above (300 mg, 1.07 mmol) in THF (5 mL) were added TEA (0.3 mL, 2.1 mmol) and 4-bromobutyryl chloride (140 pL, 1.18 mmol) at 0 °C. The reaction mixture was stirred at RT for 1 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was dissolved in THF (5 mL), and NaH (64.2 mg, 1.61 mmol, 60% in dispersion in mineral oil) was added thereto at 0 °C. The mixture was stirred at RT for 2 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title

[0960] -106-

[0961] FH13103096.12 Attorney Docket No.: LCH-03225 compound as a colorless oil (320 mg, 86%).

[0962] MS m / z: 349 [M+l]+

[0963] (Step 5) tert-butyl 2-(3-hydroxypropyl)-3-oxotetrahydropyridazine- 1 (2H)-carboxylate

[0964] (Int. C-42)

[0965] The title compound (227 mg, 96%) was synthesized following the procedure described in preparative example C-29 using the compound (320 mg, 0.92 mmol) obtained in (step 4).

[0966] MS m / z: 259 [M+l]+

[0967] (Preparative example C-431 6-(3-hydroxypropyl)-5,6-dihydro-7H-pyrrolo[3,4-b]pyridin-7- one (Int. C-43)

[0968] (Step 1) ethyl 3-methylpicolinate

[0969] To a solution of 3 -methylpicolinic acid (425 mg, 3.10 mmol) in EtOH (10 mL) was added SOCh (678 pL, 9.30 mmol) at 0 °C. The reaction mixture was stirred 80 °C for 18 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with saturated NaHCCL aqueous solution (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a colorless oil (361 mg), which was used for the next step without further purification.

[0970] 1H NMR (400 MHz, CDCh) 58.56 (dd, J = 4.6, 1.1 Hz, 1H), 7.61 (m, 1H), 7.33 (dd, J = 7.8, 4.6 Hz, 1H), 4.46 (q, J = 7.1 Hz, 2H), 2.58 (s, 3H), 1.44 (t, J = 7.1 Hz, 3H).

[0971] (Step 2) ethyl 3-(bromomethyl)picolinate

[0972] To a mixture of the compound prepared in (step 1) above (361 mg, 2.19 mmol) and AIBN (538 mg, 3.28 mmol) dissolved in chloroform (5 mL) was added NBS (583 mg, 3.28 mmol) portionwise with stirring. The reaction mixture was stirred at 80 °C for 4 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 80) to obtain the title compound as a brown oil (270 mg, 51%).

[0973] ’ H NMR (400 MHz, CDCh) 5 8.66 (dd, 7 = 4.8, 1.6 Hz, 1H), 7.87 (dd, 7 = 7.9, 1.7 Hz,

[0974] 1H), 7.45 (dd, J = 7.9, 4.8 Hz, 1H), 4.91 (s, 2H), 4.51 (q, J = 7.2 Hz, 2H), 1.47 (t, J = 7.2 Hz, -107-

[0975] FH13103096.12 Attorney Docket No.: LCH-03225

[0976] 3H).

[0977] (Step 3) 6-(3-hydroxypropyl)-5,6-dihydro-7H-pyrrolo[3,4-b]pyridin-7-one (Int. C-43)

[0978] To a solution of the compound prepared in (step 2) above (50.0 mg, 0.21 mmol) in ACN (2 mL) were added DIPEA (178 pL, 1.02 mmol) and 3-amino-l -propanol (23 pL, 0.31 mmol) at 0 °C. The reaction mixture was stirred at RT for 5 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (100 % DCM) to obtain the title compound as a white solid (31.5 mg, 80%).

[0979] ‘ H NMR (400 MHz, DMSO-&) 5 8.70 (dd, J = 4.8, 1.5 Hz, 1H), 8.05 (dd, J = 7.8, 1.5 Hz, 1H), 7.55 (dd, J = 7.8, 4.8 Hz, 1H), 4.57 - 4.52 (m, 1H), 4.50 (s, 2H), 3.63 - 3.58 (m, 2H), 3.47 > 3.43 (m, 2H), 1.81 - 1.73 (m, 2H).

[0980] [Preparative example C-441 3-((2-aminopyrimidin-4-yl)amino)propan-l-ol (Int. C-44)

[0981] (Step 1) 4-chloro-2-(methvlsulfonvl)pvrimidine

[0982] To a solution of 4-chloro-2-(methylthio)pyrimidine (500 mg, 3.11 mmol) in DCM (10 mL) was added 3-chloroperbenzoic acid (1.61 g, 9.34 mmol) at 0 °C. After stirring at RT for 2 h, the reaction was quenched by adding saturated Na2S2CL aqueous solution (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a brown solid (587 mg, 98%), which was used for the next step without further purification.

[0983] MS m / z: 193 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.82 (d, J = 5.3 Hz, 1H), 7.60 (d, 7 = 5.4 Hz, 1H), 3.38 (s, 3H).

[0984] (Step 2) 4-chloropyrimidin-2-amine

[0985] To a solution of the compound prepared in (step 1) above (200 mg, 1.04 mmol) in i- PrOH (1 mL) was added 28-30% ammonia aqueous solution (1 mL). The reaction mixture was stirred at RT for 1 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a white solid (117 mg, 87%), which was used for the next step without further purification.

[0986] MS m / z: 130 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.17 (d, J = 5.3 Hz, 1H), 6.67 (d,

[0987] -108-

[0988] FH13103096.12 Attorney Docket No.: LCH-03225

[0989] J = 5.3 Hz, 1H), 5.16 (s, 2H).

[0990] (Step 3) 3-((2-aminopyrimidin-4-yl)amino)propan-l-ol (Int. C-44)

[0991] A mixture of the compound prepared in (step 2) above (50.0 mg, 0.39 mmol) and 3- amino-1 -propanol (63.8 mg, 0.85 mmol) dissolved in EtOH (2 mL) was stirred at 80 °C for 24 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (1% TEA in MeOH : DCM = 1 : 9) to obtain the tide compound as a light brown solid (77.5 mg).

[0992] MS m / z: 169 [M+l]+. ’ H NMR (400 MHz, CD3OD) 57.53 (s, 1H), 6.03 (d, J= 6.5 Hz, 1H), 3.63 (t, J = 6.2 Hz, 2H), 3.56 - 3.44 (m, 2H), 1.89 - 1.72 (m, 2H).

[0993] [Preparative example C-45] 2-((6-aminopyridin-2-yl)oxy)ethan-l-ol (Int. C-45)

[0994] Int. C-45

[0995] (Step 1) 2-((6-bromopyridin-2-yl)oxy)ethan- 1 -ol

[0996] The title compound (132 mg, 48%) was synthesized following the procedure described in preparative example C-44 (step 3) using 2,6-dibromopyridine (300 mg, 1.26 mmol) and ethylene glycol.

[0997] MS m / z: 219 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.45 (dd, J = 8.2, 7.4 Hz, 1H), 7.09 (dd, J = 7.5, 0.8 Hz, 1H), 6.74 (dd, J = 8.2, 0.7 Hz, 1H), 4.49 - 4.41 (m, 2H), 4.00 - 3.92 (m, 2H), 2.48 (t, J = 6.0 Hz, 1H).

[0998] (Step 2) 2-((6-aminopvridin-2-vl)oxy)ethan-l-ol (Int. C-45)

[0999] To a solution of the compound prepared in (step 1) above (132 mg, 0.610 mmol) in i- PrOH (3 mL) was added 28-30% ammonia aqueous solution (4.8 mL) and copper (19.2 mg, 0.300 mmol, 200 mesh). The reaction mixture was stirred at 100 °C for 3 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 15) to obtain the title compound as a colorless oil (53.0 mg, 57%).

[1000] MS m / z: 155 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.34 (t, J = 7.8 Hz, 1H), 6.10 (d, J = 7.8 Hz, 1H), 6.06 (d, J = 7.8 Hz, 1H), 4.46 - 4.31 (m, 4H), 3.94 - 3.80 (m, 3H). [Preparative example C-461 l-(3-hydroxypropyl)pyrrolidin-2-one (Int. C-46)

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[1003] (Step 1) 1 -(3-((tetrahydro-2H-pyran-2-yl)oxy)propyl)pyrrolidin-2-one

[1004] The title compound (48.3 mg, 18%) was synthesized following the procedure described in preparative example C-35 (step 1) using 2-pyrrolidinone (100 mg, 1.18 mmol) and 2-(3- bromopropoxy)tetrahydro-2H-pyran (0.24 mL, 1.4 mmol).

[1005] 1H NMR (400 MHz, CDC13) 5 4.52 (s, 1H), 3.87 - 3.69 (m, 2H), 3.52 - 3.26 (m, 6H), 2.38 - 2.30 (m, 2H), 2.05 - 1.93 (m, 2H), 1.87 - 1.73 (m, 3H), 1.72 - 1.62 (m, 1H), 1.58 - 1.43 (m, 4H).

[1006] (Step 2) l-(3-hydroxypropyl)pyrrolidin-2-one (Int. C-46)

[1007] To a solution of the compound prepared in (step 1) above (48.3 mg, 0.21 mmol) in MeOH (3 mL) was added 1 N HC1 aqueous solution (0.5 mL). The mixture was stirred at RT for 1 h under nitrogen atmosphere. The reaction was quenched by adding saturated NaHCCL aqueous solution (20 mL), then extracted four times with 20% i-PrOH in DCM (10 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the tide compound as a colorless oil (30.0 mg, 99%).

[1008] 1H NMR (400 MHz, CDCI3) 5 3.55 - 3.46 (m, 2H), 3.44 - 3.32 (m, 4H), 2.44 - 2.35 (m, 2H), 2.07 - 1.95 (m, 2H), 1.72 - 1.62 (m, 2H).

[1009] The following intermediates (Int.C-47 ~ Int.C-50) were synthesized in the same manner as described in preparative example C-34 using corresponding amines instead of 2- pyrrolidinone.

[1010] [Preparative example C-471 3-(3-hydroxypropyl)oxazolidin-2-one (Int. C-47) [Preparative example C-481 6-(3-hydroxypropyl)-l,4,5,6-tetrahydro-7H-pyrrolo[2,3- c]pyridin-7-one (Int. C-48)

[1011] [Preparative example C-491 tert-butyl (R)-(l-(3-hydroxypropyl)-5-oxopyrrolidin-3- yl)carbamate (Int. C-49)

[1012] [Preparative example C-501 2-(3-hvdroxvpropyl)isothiazolidine 1,1-dioxide (Int. C-50)

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[1015] [Preparative example C-511 tert-butyl (1 -(2 -hydroxy ethyl)- lH-pyrazol-4-yl)carbamate (Int.

[1016] C-51)

[1017] Int. C-51

[1018] To a solution of Int. C-3 (100 mg, 0.55 mmol) in THF (5 mL) was added NaH (65.5 mg, 1.64 mmol, 60% in dispersion in mineral oil) at 0 °C. After stirring at the same temperature for 10 min, 2-bromoethanol (68.2 mg, 0.55 mmol) was added to the mixture. The reaction mixture was stirred at 60 °C for 18 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were washed with distilled water three times, dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the title compound as a white solid (86.9 mg, 70%).

[1019] 1H NMR (400 MHz, CDC13) 5 7.70 (s, 1H), 7.62 (s, 1H), 7.34 (s, 1H), 6.24 (s, 1H), 4.18 (d, J = 4.8 Hz, 2H), 3.98 (d, J = 5.0 Hz, 2H), 1.50 (s, 9H).

[1020] [Preparative example C-52] benzyl 3-((tert-butyldiphenylsilyl)oxy)bicyclo[l .1. l]pentane-l- carboxylate (Int. C-52)

[1021] Int. C-52

[1022] (Step 1) benzyl 3-hydroxybicyclo[l.l.l]pentane-l-carboxylate

[1023] To a solution of 3-hydroxybicyclo[l.l.l]pentane-l-carboxylic acid (300 mg, 2.34 mmol) in DMF (4.7 mL) was added NaHCCL (590 mg, 7.02 mmol). After stirring for 10 min, benzyl bromide (3.1 mL, 26.3 mmol) was added dropwise thereto, then the resulting mixture was stirred at RT for 20 h under nitrogen atmosphere. On completion, the reaction was quenched by adding distilled water (50 mL), then extracted three times with EA (50 mL). The combined organic layers were washed with distilled water three times, dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was

[1024] -I l l-

[1025] FH13103096.12 Attorney Docket No.: LCH-03225 purified by MPLC (EA : n-hexane = 1 : 2 to 1 : 1) to obtain the title compound as a colorless oil (492 mg, 96%).

[1026] MS m / z: 219 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.34 (q, J = 5.7 Hz, 5H), 5.12 (s, 2H), 2.49 (s, 1H), 2.23 (d, J = 1.1 Hz, 6H).

[1027] (Step 2) benzyl 3 - ((tert-butyldiphenylsilyl)oxy)bicyclo [1.1.1] pentane- 1 -carboxylate (Int. C-52)

[1028] To a solution of the compound prepared in (step 1) above (492 mg, 2.25 mmol) in DMF (7.5 mL) were added imidazole (368 mg, 5.41 mmol) and tert-butyldiphenylsilyl chloride (0.69 mL, 2.70 mmol). The reaction mixture was stirred at RT for 19 h under nitrogen atmosphere. On completion, the residue was diluted with distilled water (50 mL), then extracted three times with EA (50 mL). The combined organic layers were washed with saturated NaHCO t aqueous solution, dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 5 : 95) to obtain the title compound as a colorless oil (870 mg, 85%).

[1029] 1H NMR (400 MHz, CDCh) 5 7.74 - 7.66 (m, 4H), 7.46 - 7.35 (m, 6H), 7.31 (d, J = 7.8 Hz, 3H), 7.24 (d, J = 5.6 Hz, 2H), 5.01 (s, 2H), 1.98 (d, J = 1.4 Hz, 6H), 1.01 (d, J = 1.4 Hz, 9H).

[1030] [Preparative example C-531 tert-butyl (3-(3-hydroxypropyl)bicyclo[l .1. l]pentan-l- yl)carbamate (Int. C-53)

[1031] (Step 1) tert-butyl (3-(hydroxymethyl)bicyclo [1.1.1 ]pentan- 1 -yl)carbamate

[1032] To a solution of 3-((tert-butoxycarbonyl)amino)bicyclo[l.l.l]pentane-l-carboxylic acid (300 mg, 1.32 mmol) in THF (7 mL) was added 1 M BHyTHF complex solution in THF (2.6 mL, 2.6 mmol) at 0 °C, then the reaction mixture was stirred at RT for 24 h under nitrogen atmosphere. On completion, MeOH (1.4 mL) was added slowly, then the reaction mixture was concentrated under reduced pressure. The residue was diluted with saturated NaHCOt aqueous solution (30 mL), then extracted with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the title compound as a white solid (230 mg, 82%).

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[1035] MS m / z: 214 [M+l]+.1H NMR (400 MHz, CDCh) 5 4.95 (s, 1H), 3.71 (d, J = 6.3 Hz, 2H), 1.94 (s, 6H), 1.45 (s, 9H), 1.20 (t, J = 6.4 Hz, 1H).

[1036] (Step 2) tert-butyl (3-formylbicyclo [1.1. l]pentan- 1 -yl)carbamate

[1037] To a solution of oxalyl chloride (0.14 mL, 1.62 mmol) in anhydrous DCM (5 mL) was added DMSO (0.23 mL, 3.24 mmol) at -70 °C. After stirring at the same temperature for 30 min, the compound prepared in (step 1) above (230 mg, 1.08 mmol) dissolved in anhydrous DCM (5 mL) was added thereto. After stirring at the same temperature for 1 h, TEA (0.90 mL, 6.47 mmol) was added, then the resulting mixture was stirred at RT for 1 h. On completion, the reaction mixture was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The tide compound was obtained as a light brown solid (255 mg), which was used for the next step without further purification.

[1038] MS m / z: 212 [M+l]+.1H NMR (400 MHz, CDCh) 59.66 (s, 1H), 4.99 (s, 1H), 2.28 (s, 6H), 1.45 (s, 9H).

[1039] (Step 3) ethyl (E)-3-(3-((tert-butoxycarbonyl)amino)bicyclo[l.l.l]pentan-l-yl)acrylate

[1040] To a solution of the compound prepared in (step 2) above (228 mg, 1.08 mmol) in anhydrous THF (10 mL) was added (carbethoxymethylene)triphenyl phosphorane (751 mg, 2.16 mmol). The reaction mixture was stirred at RT for 18 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 5) to obtain the title compound as a white solid (241 mg, 79%).

[1041] MS m / z: 282 [M+l]+.1H NMR (400 MHz, CDCh) 5 6.99 (d, J = 15.6 Hz, 1H), 5.78 (d, J = 15.6 Hz, 1H), 4.97 (s, 1H), 4.18 (q, J = 7.1 Hz, 2H), 2.12 (s, 6H), 1.44 (s, 9H), 1.28 (t, J = 7.1 Hz, 3H).

[1042] (Step 4) ethyl 3-(3-((tert-butoxycarbonyl)amino)bicyclo [1.1.1 Jpentan- 1 -yl)propanoate

[1043] To a solution of the compound prepared in (step 3) above (241 mg, 0.86 mmol) in a mixed solvent of MeOH / EA (8 mL, 1:1) was added Pd / C (48.0 mg). After stirring at RT for 17 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to obtain the title compound as a colorless oil (234 mg, 96%), which was used for the next step without further purification.

[1044] MS m / z: 284 [M+l]+. ’ H NMR (400 MHz, CDCh) 54.90 (s, 1H), 4.14 - 4.09 (m, 2H), 2.27 (t, J = 7.7 Hz, 2H), 1.92 - 1.77 (m, 8H), 1.43 (s, 9H), 1.27 - 1.23 (m, 3H).

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[1047] (Step 5) tert-butyl (3-(3-hydroxypropyl)bicyclo[l.l.l]pentan-l-yl)carbamate (Int. CI- 53)

[1048] To a solution of the compound prepared in (step 4) above (234 mg, 0.83 mmol) in anhydrous THF (6 mL) was added LiBF (89.9 mg, 4.13 mmol) at 0 °C. After stirring at RT for 23 h under nitrogen atmosphere, the reaction was quenched by adding distilled water (20 mL), then extracted with EA (20 mL). The organic layer was dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 2 : 3) to obtain the tide compound as a white solid (184 mg, 92%).

[1049] MS m / z: 242 [M+l]+.1H NMR (400 MHz, CDCh) 54.91 (s, 1H), 3.67 - 3.60 (m, 2H), 1.85 (s, 6H), 1.62 - 1.58 (m, 2H), 1.55 - 1.48 (m, 2H), 1.44 (s, 9H), 1.26 - 1.22 (m, 1H). (Preparative example C-541 tert-butyl (4-(3 -hydroxypropyl)bicy clo [2.1.1 ]hexan- 1 - yl)carbamate (Int. C-54)

[1050] Int. C-54

[1051] The title compound was synthesized in the same manner as described in preparative example C-53 using 4-((tert-butoxycarbonyl)amino)bicyclo[2.1.1] hexane- 1 -carboxylic acid.

[1052] MS m / z: 256 [M+l]+.1H NMR (400 MHz, CDCh) 5 4.89 (s, 1H), 3.63 (s, 2H), 1.80 - 1.75 (m, 2H), 1.74 - 1.65 (m, 2H), 1.54 - 1.47 (m, 3H), 1.44 (s, 10H), 1.40 - 1.37 (m, 2H), 1.29 - 1.22 (m, 2H).

[1053] (Preparative example C-551 3-(3-((tert-butyldiphenylsilyl)oxy)bicyclo[l .1. l]pentan-l- yl)propan-l-ol (Int. C-55)

[1054] Int. C-55

[1055] The title compound was synthesized in the same manner as described in preparative example C-53 using benzyl 3-((tert-butyldiphenylsilyl)oxy) bicy clo [1.1.1 ]pentane-l- carboxylate.

[1056] MS m / z: 381 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.76 - 7.68 (m, 4H), 7.47 - 7.32 (m, 6H), 3.53 (q, J = 5.8 Hz, 2H), 1.53 (s, 6H), 1.47 - 1.30 (m, 4H), 1.00 (s, 9H).

[1057] [Preparative example C-561 3-(((tert-butyldiphenylsilyl)oxy)methyl)-l-(3- hydroxypropyl)piperidin-2-one (Int. C-56)

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[1060] (Step 1) 3-(hydroxymethyl)piperidin-2-one

[1061] To a solution of 3-ethoxycarbonyl-2-piperidone (500 mg, 2.92 mmol) in anhydrous MeOH (6 mL) were added CaCh (357 mg, 3.21 mmol) and NaBt (243 mg, 6.43 mmol) at 0 °C. The mixture was stirred at RT for 4 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with 1 N HC1 aqueous solution (50 mL), then extracted twice with 25% i-PrOH in chloroform (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a colorless oil (458 mg), which was used for the next step without further purification.

[1062] MS m / z: 130 [M+l]+.1H NMR (400 MHz, CDCh) 55.86 (s, 1H), 4.07 - 4.03 (m, 1H), 3.79 - 3.62 (m, 2H), 3.40 - 3.21 (m, 2H), 2.55 - 2.43 (m, 1H), 1.95 - 1.80 (m, 3H), 1.53 - 1.45 (m, 1H).

[1063] (Step 2) 3-(((tert-butyldiphenylsilyl)oxy)methyl)piperidin-2-one

[1064] To a solution of the compound prepared in (step 1) above (377 mg, 2.92 mmol) in DMF (7 mL) were added imidazole (239 mg, 3.50 mmol) and tert-butyldiphenylchlorosilane (0.83 mL, 3.21 mmol). The reaction mixture was stirred at RT for 20 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (20 mL), then extracted with EA (20 mL). The organic layer was washed with distilled water (20 mL), dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 2 : 1) to obtain the title compound as a white solid (422 mg, 39%).

[1065] MS m / z: 368 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.70 - 7.62 (m, 4H), 7.45 - 7.33 (m, 6H), 5.84 (s, 1H), 4.03 (dd, J = 9.8, 6.6 Hz, 1H), 3.92 (dd, J = 9.8, 3.6 Hz, 1H), 3.34 - 3.26 (m, 2H), 2.56 - 2.47 (m, 1H), 2.05 - 2.00 (m, 1H), 1.97 - 1.87 (m, 2H), 1.79 - 1.68 (m, 1H), 1.05 (s, 9H).

[1066] (Step 3) ethyl 3-(3-(((tert-butyldiphenylsilyl)oxy)methyl)-2-oxopiperidin- 1- yl)propanoate

[1067] To a solution of the compound prepared in (step 2) above (328 mg, 0.89 mmol) in toluene (2.5 mL) were added CsF (13.5 mg, 0.09 mmol), tetraethoxysilane (0.89 mL), and ethyl acrylate (0.89 mL). The reaction mixture was stirred at RT for 72 h under nitrogen atmosphere.

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[1070] On completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 10) to obtain the title compound as a colorless oil (164 mg, 39%).

[1071] MS m / z: 468 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.69 - 7.63 (m, 4H), 7.43 - 7.33 (m, 6H), 4.11 (q, J = 7.0 Hz, 2H), 4.02 (dd, J = 9.8, 6.5 Hz, 1H), 3.90 (dd, J = 9.7, 3.4 Hz, 1H), 3.68 - 3.52 (m, 2H), 3.40 - 3.29 (m, 2H), 2.59 (t, J = 7.3 Hz, 2H), 2.52 - 2.44 (m, 1H), 2.04 - 1.86 (m, 3H), 1.79 - 1.66 (m, 1H), 1.23 (t, J = 7.1 Hz, 3H), 1.04 (s, 9H).

[1072] (Step 4) 3-(((tert-butyldiphenylsilyl)oxy)methyl)- 1 -(3-hydroxypropyl)piperidin-2-one (Int. C-56)

[1073] The title compound (87.2 mg, 58%) was synthesized following the procedure described in preparative example C-53 (step 5) using the compound (164 mg, 0.35 mmol) obtained in (step 3) above.

[1074] MS m / z: 426 [M+l]+

[1075] [Preparative example C-571 l-(3-aminopropyl)pyrrolidin-2-one (Int. C-57)

[1076] (Step 1) 2-(3-(2-oxopyrrolidin- 1 -yl)propyl)isoindoline- 1 ,3-dione

[1077] To a mixture of Int. C-46 (100 mg, 0.430 mmol) and phthalimide (46.2 mg, 0.31 mmol) dissolved in THF (4 mL) were added PPI13 (82.4 mg, 0.31 mmol) and DIAD (61.9 pL, 0.31 mmol) at 0 °C sequentially. The mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : DCM = 2 : 3) to obtain the title compound as a colorless oil (44.1 mg).

[1078] MS m / z: 273 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.87 - 7.80 (m, 2H), 7.74 - 7.69 (m, 2H), 3.74 - 3.66 (m, 2H), 3.47 - 3.33 (m, 4H), 2.43 - 2.34 (m, 2H), 2.10 - 2.00 (m, 2H), 1.98 - 1.88 (m, 2H).

[1079] (Step 2) l-(3-aminopropyl)pyrrolidin-2-one (Int. C-57)

[1080] The title compound (31.9 mg) was synthesized following the procedure described in preparative example C-18 (step 2) using the compound (44.1 mg) obtained in (step 1) above.

[1081] ‘ H NMR (400 MHz, CDCh) 5 3.51 - 3.47 (m, 2H), 3.40 - 3.35 (m, 4H), 2.71 - 2.65 (m, 2H), 2.42 - 2.36 (m, 2H), 2.07 - 2.00 (m, 2H), 1.84 - 1.76 (m, 2H).

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[1084] The following intermediates (Int.C-58 ~ Int. C-69) were synthesized in the same manner as described in preparative example C-57 using the corresponding alcohols instead of Int. C-46.

[1085] [Preparative example C-581 3-(3-aminopropyl)oxazolidin-2-one (Int. C-58)

[1086] [Preparative example C-591 2-(3-aminopropvl)isothiazolidine 1,1 -dioxide (Int. C-59)

[1087] [Preparative example C-601 tert-butyl (l-(3-amino-2,2-difluoropropyl)-lH-pyrazol-4- yl)carbamate (Int. C-60)

[1088] [Preparative example C-611 tert-butyl 2-(3-aminopropyl)-3-oxotetrahydropyridazine- 1 (2H)- carboxylate (Int. C-61)

[1089] [Preparative example C-62] tert-butyl 4-(3-aminopropyl)-3-oxopiperazine-l-carboxylate

[1090] (Int. C-62)

[1091] [Preparative example C-631 N4-(3-aminopropyl)pyrimidine-2,4-diamine (Int. C-63)

[1092] [Preparative example C-64] 6-(2-aminoethoxy)pyridin-2-amine (Int. C-64)

[1093] [Preparative example C-651 6-(3-aminopropyl)-5,6-dihydro-7H-pyrrolo[3,4-b]pyridin-7-one

[1094] (Int. C-65)

[1095] [Preparative example C-661 6-(3-aminopropvl)-l,4,5.6-tetrahvdro-7H-pvrrolo[2,3-c1pyridin-

[1096] 7-one (Int. C-66)

[1097] [Preparative example C-671 tert-butyl (R)-(l-(3-aminopropyl)-5-oxopyrrolidin-3- yl)carbamate (Int. C-67)

[1098] [Preparative example C-681 tert-butyl (3-(3-aminopropyl)bicyclo[l .1. l]pentan-l- yl)carbamate (Int. C-68)

[1099] [Preparative example C-691 tert-butyl (4-(3 -aminopropyl)bicyclo [2.1.1 ]hexan- 1 - yl)carbamate (Int. C-69)

[1100] FH13103096.12 Attorney Docket No.: LCH-03225

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[1103] (Step 1) 2-(3-(3-(((tert-butyldiphenylsilyl)oxy)methyl)-2-oxopiperidin-l- yl)propyl)isoindoline- 1 ,3-dione

[1104] The title compound (41.4 mg, 36%) was synthesized following the procedure described in preparative example C-57 (step 1) using Int.C-56 (87.2 mg, 0.21 mmol).

[1105] MS m / z: 555 [M+l]+

[1106] (Step 2) 2-(3-(3-(hvdroxvmethvl)-2-oxopiperidin-l-vl)propyl)isoindoline-1.3-dione

[1107] To a solution of the compound prepared in (step 1) above (41.4 mg, 0.07 mmol) in anhydrous THF (1 mL) was added 1 M TBAF solution in THF (0.11 mL) at 0 °C. The mixture was stirred at RT for 3 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the title compound as a light brown oil (18.0 mg, 76%).

[1108] MS m / z: 317 [M+l]+

[1109] (Step 3) l-(3-aminopropyl)-3-(hydroxymethyl)piperidin-2-one (Int. C-70)

[1110] The title compound (10.9 mg) was synthesized following the procedure described in preparative example C-18 (step 2) using the compound (18.0 g, 0.06 mmol) obtained in (step 2) above.

[1111] MS m / z: 187 [M+l]+

[1112] (Preparative example C-711 3-(3-aminopropyl)bicyclo[l.l.l]pentan-l-ol (Int. C-71)

[1113] Int. C-71

[1114] The title compound was synthesized in the same manner as described in preparative example C-70 using Int. C-55.

[1115] MS m / z: 142 [M+l]+. ’ H NMR (400 MHz, CDC13) 5 3.65 (q, 7 = 6.0 Hz, 1H), 2.65 (t, 7 = 7.1 Hz, 2H), 1.72 (s, 6H), 1.55 - 1.52 (m, 2H), 1.39 (p, 7 = 7.5 Hz, 2H).

[1116] (Preparative example C-721 tert-butyl (3-(2-aminoethyl)bicyclo[l.l.l]pentan-l- yl)carbamate (Int. C-72)

[1117] (Step 1) (3-((tert-butoxycarbonyl)amino)bi cyclo [1.1.1 Jpentan- 1 -yl)methyl methanesulfonate

[1118] To a solution of tert-butyl (3-(hydroxymethyl)bicyclo[l.l.l]pentan-l-yl)carbamate which was synthesized in preparative example C-53 (step 1) (150 mg, 0.70 mmol) in DCM (4 -119-

[1119] FH13103096.12 Attorney Docket No.: LCH-03225 mL) were added TEA (0.15 mL, 1.06 mmol) and methanesulfonyl chloride (65.0 pL, 0.84 mmol) at 0 °C. The mixture was stirred at RT for 1 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (20 mL), then extracted with DCM (20 mL). The organic layer was dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The title compound was obtained as a white solid (196 mg, 96%), which was used for the next step without further purification.

[1120] MS m / z: 292 [M+l]+.1H NMR (400 MHz, CDC13) 54.97 (s, 1H), 4.30 (s, 2H), 3.01 (s, 3H), 2.04 (s, 6H), 1.44 (s, 9H).

[1121] (Step 2) tert-butyl (3-(cyanomethyl)bicyclo [1.1.1 Jpentan- 1 -yl)carbamate

[1122] A mixture of the compound prepared in (step 1) above (196 mg, 0.67 mmol) and NaCN (98.9 mg, 2.02 mmol) dissolved in DMSO (6 mL) was stirred at 100 °C for 1 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (20 mL), then extracted with EA (20 mL). The organic layer was washed with distilled water (20 mL), dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 3) to obtain the title compound as a white solid (123 mg, 82%).

[1123] MS m / z: 223 [M+l]+.1H NMR (400 MHz, CDC13) 54.95 (s, 1H), 2.65 (s, 2H), 2.06 (s, 6H), 1.44 (s, 9H).

[1124] (Step 3) tert-butyl (3-(2-aminoethyl)bicyclo[l.l.l]pentan-l-yl)carbamate (Int. C-72)

[1125] To a solution of the compound prepared in (step 2) above (123 mg, 0.55 mmol) in a mixed solvent of MeOH / THF (8 mL, 3 : 1) were added Raney-Ni (123 mg) and 28-30% ammonia aqueous solution (0.4 mL). After stirring at RT for 17 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to obtain the title compound as a gray oil (143 mg), which was used for the next step without further purification.

[1126] MS m / z: 227 [M+l]+[Preparative example C-731 tert-butyl (3-(4-aminobutyl)bicyclo [1.1.1 Jpentan- 1 - yl)carbamate (Int. C-73)

[1127] The title compound was synthesized in the same manner as described in preparative example C-72 using Int. C-53.

[1128] MS m / z: 255 [M+l]+

[1129] -120-

[1130] FH13103096.12 Attorney Docket No.: LCH-03225

[1131] [Preparative example C-74] l-amino-5-(3-aminopropyl)pyridin-2(lH)-one (Int. C-74)

[1132] (Step 1) ethyl (E)-3-(6-methoxypyridin-3-yl)acrylate

[1133] A mixture of 5-bromo-2-methoxypyridine (1.00 g, 5.32 mmol), ethyl acrylate (0.87 mL, 7.98 mmol), Pd(0Ac)2 (119 mg, 0.53 mmol), DIPEA (1.85 mL, 10.6 mmol) and tris(o- tolyl)phosphine (324 mg, 1.06 mmol) dissolved in ACN (10 mL) was stirred at 90 °C for 17 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 9) to obtain the tide compound as a light-yellow solid (1.11 g, 100%).

[1134] MS m / z: 208 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.28 (s, 1H), 7.80 - 7.75 (m, 1H), 7.63 (d, J = 16.0 Hz, 1H), 6.77 (d, J = 8.8 Hz, 1H), 6.34 (d, J = 16.0 Hz, 1H), 4.31 - 4.23 (m, 2H), 3.97 (s, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[1135] (Step 2) ethyl 3-(6-methoxypyridin-3-yl)propanoate

[1136] To a solution of the compound prepared in (step 1) above (1.11 g, 5.37 mmol) in EtOH (54 mL) was added Pd / C (200 mg). After stirring at RT for 14 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to obtain the title compound as a yellow solid (1.10 g, 97%), which was used for the next step without further purification.

[1137] MS m / z: 210 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.00 (s, 1H), 7.46 - 7.40 (m, 1H), 6.68 (d, J = 8.5 Hz, 1H), 4.17 - 4.08 (m, 2H), 3.91 (s, 3H), 2.87 (t, J = 7.6 Hz, 2H), 2.58 (t, J = 7.6 Hz, 2H), 1.24 (t, J = 7.1 Hz, 3H).

[1138] (Step 3) 3-(6-methoxypyridin-3-yl)propan-l-ol

[1139] The title compound (803 mg, 89%) was synthesized following the procedure described in preparative example C-41 (step 1) using the compound (1.11 g, 8.37 mmol) obtained in (step 2) above.

[1140] FH13103096.12 Attorney Docket No.: LCH-03225

[1141] MS m / z: 168 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.99 (s, 1H), 7.46 - 7.40 (m, 1H), 6.69 (d, J = 8.5 Hz, 1H), 3.92 (s, 3H), 3.71 - 3.64 (m, 2H), 2.64 (t, J = 7.7 Hz, 2H), 1.90 - 1.81 (m, 2H), 1.36 - 1.30 (m, 1H).

[1142] (Step 4) 2-(3-(6-methoxypyridin-3-yl)propyl)isoindoline- 1 ,3-dione

[1143] The title compound (1.20 g) was synthesized following the procedure described in preparative example C-57 (step 1) using the compound (400 mg, 2.39 mmol) obtained in (step 3) above.

[1144] MS m / z: 297 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.00 - 7.95 (m, 1H), 7.86 - 7.82 (m, 2H), 7.74 - 7.68 (m, 2H), 7.45 - 7.41 (m, 1H), 6.66 (d, J = 8.5 Hz, 1H), 3.88 (s, 3H), 3.74 (t, J = 7.1 Hz, 2H), 2.61 (t, . / = 7.7 Hz, 2H), 2.04 - 1.95 (m, 2H).

[1145] (Step 5) 2-(3-(6-hvdroxvpvridin-3-vl)propyl)isoindoline-l.3-dione

[1146] A solution of the compound prepared in (step 4) above (540 mg) in 33 wt% HBr / acetic acid (10 mL) was stirred at 100 °C for 3 h under nitrogen atmosphere. On completion, the reaction mixture was cooled to RT, diluted with distilled water (100 mL), then extracted twice with 10% MeOH in DCM (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 19) to obtain the title compound as a white solid (414 mg, 80%).

[1147] MS m / z: 283 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.87 - 7.82 (m, 2H), 7.75 - 7.69 (m, 2H), 7.38 - 7.33 (m, 1H), 7.16 (s, 1H), 6.54 (d, J = 9.5 Hz, 1H), 3.72 (t, J = 6.8 Hz, 2H), 2.44 (t, J = 7.7 Hz, 2H), 1.99 - 1.88 (m, 2H).

[1148] (Step 6) 2-(3-(l-amino-6-oxo-1.6-dihvdropvridin-3-vl)propvl)isoindoline-1.3-dione

[1149] A mixture of the compound prepared in (step 5) above (200 mg, 0.71 mmol), CS2CO3 (462 mg, 1.42 mmol), and O-diphenylphosphorylhydroxylamine (248 mg, 1.06 mmol) dissolved in DMF (3.5 mL) was stirred at RT for 3 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with 10% MeOH in DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 19) to obtain the title compound as a light brown oil (136 mg, 65%).

[1150] MS m / z: 298 [M+l]+. ’ H NMR (400 MHz, DMSO-A) 5 7.87 - 7.82 (m, 4H), 7.60 (s, 1H), 7.30 (d, J = 9.1 Hz, 1H), 6.39 (d, J = 9.1 Hz, 1H), 6.06 (s, 2H), 3.58 (t, J = 6.2 Hz, 2H), 2.38 (t, J = 8.2 Hz, 2H), 1.83 - 1.76 (m, 2H).

[1151] -122-

[1152] FH13103096.12 Attorney Docket No.: LCH-03225

[1153] (Step 7) l-amino-5-(3-aminopropyl)pyridin-2(lH)-one (Int. C-74)

[1154] The title compound (22.6 mg, 80%) was synthesized following the procedure described in preparative example C-18 (step 2) using the compound (50.0 mg, 0.17 mmol) obtained in (step 6).

[1155] MS m / z: 168 [M+l]+.1H NMR (400 MHz, DMSO-rfe) 5 7.53 (s, 1H), 7.27 (d, J = 9.1 Hz, 1H), 6.40 (d, J = 8.9 Hz, 1H), 6.08 (s, 2H), 2.38 - 2.33 (m, 2H), 1.57 - 1.49 (m, 2H). [Preparative example C-751 2-((tetrahydro-2H-pyran-2-yl)oxy)ethan-l -amine (Int. C-75)

[1156] (Step 1) N,N-dibenzyl-2-((tetrahydro-2H-pyran-2-yl)oxy)ethan- 1 -amine

[1157] To a solution of 2-(dibenzylamino)ethan-l-ol hydrochloride (500 mg, 1.80 mmol) in DCM (85 mL) was added 3,4-dihydro-2H-pyran (0.240 mL, 2.70 mmol). The reaction mixture was stirred at RT for 14 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (50 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n- hexane = 1 : 9) to obtain the title compound as a colorless oil (452 mg, 77%).

[1158] MS m / z: 326 [M+l]+

[1159] (Step 2) 2-((tetrahydro-2H-pvran-2-vl)oxy)ethan-l -amine (Int. C-75)

[1160] The title compound (94.0 mg) was synthesized following the procedure described in preparative example C-29 using the compound (211 mg, 0.65 mmol) obtained in (step 1) above.

[1161] MS m / z: 146 [M+l]+

[1162] [Preparative example C-761 tert-butyl (5-bromo-2-chloropyrimidin-4-yl)carbamate (Int. C- 76)

[1163] Int. C-76

[1164] The title compound (304 mg, 82%) was synthesized following the procedure described in preparative example C-40 (step 1) using 5-bromo-2-chloropyrimidin-4-amine (250 mg, 1.20 mmol).

[1165] MS m / z: 308 [M]+. ’ H NMR (400 MHz, CDCh) 5 8.48 (s, 1H), 7.44 (s, 1H), 1.56 (s, 9H).

[1166] [Preparative example C-77] benzyl (3-((2-chloropyrimidin-4-yl)amino)propyl)carbamate

[1167] (Int. C-77)

[1168] -123-

[1169] FH13103096.12 Attorney Docket No.: LCH-03225

[1170] The title compound (132 mg, 61%) was synthesized following the procedure described in preparative example C-44 (step 3) using 2,4-dichloropyrimidine (100 mg, 0.67 mmol).

[1171] MS m / z: 321 [M+l]

[1172] [Preparative example C-781 tert-butyl ( 1 -(3-aminopropyl)-2-oxopiperidin-3-yl)carbamate

[1173] Int. C-78

[1174] (Step 1) tert-butyl (l-(3-(((benzyloxy)carbonyl)amino)propyl)-2-oxopiperidin-3- yl)carbamate

[1175] The tide compound (110 mg, 38%) was synthesized following the procedure described in preparative example C-35 (step 1) using Int. C-14 (150 mg, 0.70 mmol).

[1176] MS m / z: 406 [M+l]+

[1177] (Step 2) tert-butyl (l-(3-aminopropyl)-2-oxopiperidin-3-yl)carbamate (Int. C-78)

[1178] The title compound (70.0 mg, 77%) was synthesized following the procedure described in preparative example C-29 using the compound (118 mg, 0.29 mmol) obtained in (step 1) above.

[1179] MS m / z: 272 [M+l]+

[1180] The following intermediates (Int.C-79 ~ Int.C-81) were synthesized in the same manner as described in preparative example C-78 using the corresponding amines instead of Int. C-14.

[1181] [Preparative example C-791 2-(3-aminopropyl)-8-oxa-2-azaspiro[4.5]decan-l-one (Int. C-79) [Preparative example C-801 2-(3-aminopropyl)-8-oxa-2-azaspiro[4.5]decan-3-one (Int. C-80) [Preparative example C-811 tert-butyl 5-(3-aminopropyl)-4-oxo-4,5,6,7-tetrahydro-lH- pyrrolo[3,2-c]pyridine-l-carboxylate (Int. C-81)

[1182] -124-

[1183] FH13103096.12 Attorney Docket No.: LCH-03225

[1184] (Int. C-82)

[1185] (Step 1) tert-butyl (2-((3-(((benzyloxy)carbonyl)amino)propyl)amino)-5- bromopyrimidin-4-yl)carbamate

[1186] A mixture of Int. C-76 (100 mg, 0.32 mmol), TEA (90 pL, 0.65 mmol), and benzyl N- (3-aminopropyl)carbamate (74.2 mg, 0.36 mmol) dissolved in EtOH (2 mL) was stirred at 80 °C for 4 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 3 : 7 to 1 : 1) to obtain the title compound as a cloudy oil (40.0 mg, 26%).

[1187] MS m / z: 481 [M+l]+

[1188] (Step 2) tert-butyl (2-((3-aminopropyl)amino)pyrimidin-4-yl)carbamate (Int. C-82)

[1189] The title compound (21.0 mg) was synthesized following the procedure described in preparative example C-29 using the compound (40.0 mg, 0.08 mmol) obtained in (step 1) above.

[1190] MS m / z: 346 [M+l]+

[1191] [Preparative example C-831 N-(pyrazin-2-yl)propane-l,3-diamine (Int. C-83)

[1192] Int. C-83

[1193] The title compound was synthesized in the same manner as described in preparative example C-82 using 2-chloropyrazine.

[1194] MS m / z: 153 [M+l]+

[1195] [Preparative example C-841 N4-(3-aminopropyl)-N2-(pyridin-2-yl)pyrimidine-2,4-diamine

[1196] (Int. C-84)

[1197] FH13103096.12 Attorney Docket No.: LCH-03225

[1198] (Step 1) benzyl (3-((2-(pyridin-2-ylamino)pyrimidin-4-yl)amino)propyl)carbamate

[1199] A mixture of Int. C-77 (80.0 mg, 0.25 mmol), 2-aminopyridine (23.5 mg, 0.25 mmol), xantphos (28.9 mg, 0.05 mmol), and Pd2(dba)3 (228 mg, 0.25 mmol), and CS2CO3 (122 mg, 0.37 mmol) dissolved in 1.4-dioxane (2 mL) was stirred at 100 °C for 3 h under nitrogen atmosphere. On completion, the reaction mixture was cooled to RT, filtered through a Celite pad, washed with EA (50 mL). The filtrate was diluted with distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 2 : 1 to 0 : 1) to obtain the title compound as an ivory solid (42.0 mg, 45%).

[1200] MS m / z: 379 [M+l]+

[1201] (Step 2) N4-(3-aminopropyl)-N2-(pyridin-2-yl)pyrimidine-2,4-diamine (Int. C-84)

[1202] The tide compound (19.0 mg, %) was synthesized following the procedure described in preparative example C-29 using the compound (42.0 mg, 0.11 mmol) obtained in (step 1) above.

[1203] MS m / z: 245 [M+l]+

[1204] [Preparative example C-851 N4-(3-aminopropyl)-N2-(2-((tetrahydro-2H-pyran-2- yl)oxy)ethyl)pyrimidine-2,4-diamine (Int. C-85)

[1205] Int. C-85

[1206] The title compound was synthesized in the same manner as described in preparative example C-84 using Int. C-77 and Int. C-75.

[1207] MS m / z: 296 [M+l]+

[1208] [Preparative example C-861 4-((3-aminopropyl)amino)-l,3,5-triazin-2-ol (Int. C-86)

[1209] (Step 1) benzyl (3-((4,6-dichloro-l,3,5-triazin-2-yl)amino)propyl)carbamate

[1210] To a solution of cyanuric chloride (80.0 mg, 0.43 mmol) in EA (2 mL) were added DIPEA (75.6 pL, 0.43 mmol) and benzyl(3-aminopropyl)carbamate (90.3 mg, 0.43 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 20 min under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, -126-

[1211] FH13103096.12 Attorney Docket No.: LCH-03225 then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 2) to obtain the title compound as a light brown solid (77.2 mg, 50%).

[1212] MS m / z: 357 [M+l]+. ’ H NMR (400 MHz, CDC13) 5 7.40 - 7.27 (m, 5H), 7.04 (t, . / = 6.9 Hz, 1H), 5.20 - 5.14 (m, 1H), 5.10 (s, 2H), 3.56 - 3.46 (m, 2H), 3.34 - 3.19 (m, 2H), 1.81 - 1.69 (m, 2H).

[1213] (Step 2) benzyl (3-((4-chloro-6-hydroxy-l,3,5-triazin-2-yl)amino)propyl)carbamate

[1214] To a solution of the compound prepared in (step 1) above (77.2 mg, 0.22 mmol) in a mixed solvent of EA / acetone (1 mL, 1 : 1) was added NaOH (9.54 mg, 0.24 mmol) dissolved in distilled water (1 mL). The mixture was stirred at RT for 3 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (10 mL), then extracted twice with EA (10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 9) to obtain the title compound as a colorless oil (15.0 mg, 20%).

[1215] MS m / z: 338 [M+l]+.1H NMR (400 MHz, CDCh) 59.22 (s, 1H), 7.36 - 7.27 (m, 6H), 5.52 (s, 1H), 5.08 (s, 2H), 3.58 - 3.46 (m, 2H), 3.28 - 3.18 (m, 2H), 1.88 - 1.76 (m, 2H).

[1216] (Step 3) 4-((3-aminopropyl)amino)-L3.5-triazin-2-ol (Int. C-86)

[1217] The title compound (4.60 mg) was synthesized following the procedure described in preparative example C-29 using the compound (15.0 mg, 0.044 mmol) obtained in (step 2) above.

[1218] MS m / z: 304 [M+l]+[Preparative example C-87] l-(3-aminopropyl)-lH-pyrazole-4-carboxylic acid (Int. C-87)

[1219] (Step 1) ethyl l-(3-(((benzyloxy)carbonyl)amino)propyl)-lH-pyrazole-4-carboxylate

[1220] To a mixture of benzyl N-(3-bromopropyl)carbamate (100 mg, 0.37 mmol) and K2CO3 (66.0 mg, 0.48 mmol) dissolved in anhydrous DMF (3.00 mL) was added ethyl pyrazole-4- carboxylate (56.6 mg, 0.40 mmol). The reaction mixture was stirred at 80 °C for 2 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (20 mL), then extracted with EA (20 mL). The organic layer was washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting

[1221] -127-

[1222] FH13103096.12 Attorney Docket No.: LCH-03225 residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the title compound as a colorless oil (108 mg, 89%).

[1223] MS m / z: 332 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.93 - 7.86 (m, 2H), 7.38 - 7.32 (m, 5H), 5.10 (s, 2H), 5.02 (s, 1H), 4.28 (q, J = 7.1 Hz, 2H), 4.20 (t, J = 6.6 Hz, 2H), 3.20 (q, J = 6.3 Hz, 2H), 2.12 - 2.03 (m, 2H), 1.34 (t, J = 7.1 Hz, 3H).

[1224] (Step 2) l-(3-(((benzyloxy)carbonyl)amino)propyl)-lH-pyrazole-4-carboxylic acid

[1225] To a solution of the compound prepared in (step 1) above (108 mg, 0.33 mmol) in a mixed solvent of THF / MeOH / distilled water (4.2 mL, 15 : 1 : 5) was added LiOH monohydrate (27.4 mg, 0.65 mmol). The reaction mixture was stirred at 60 °C for 2 h under nitrogen atmosphere. After being cooled to RT, the reaction was quenched by adding 1 N HC1 aqueous solution (20 mL), then extracted with EA (20 mL). The organic layer was dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a colorless oil (102 mg), which was used for the next step without further purification.

[1226] MS m / z: 304 [M+l]+

[1227] (Step 3) l-(3-aminopropyl)-lH-pyrazole-4-carboxylic acid (Int. C-87)

[1228] To a solution of the compound prepared in (step 2) above (102 mg, 0.34 mmol) in a mixed solvent of MeOH / EA (3 mL, 1 : 1) was added Pd / C (20 mg). After stirring at RT for 15 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to obtain the title compound as a white solid (32.0 mg, 55%), which was used for the next step without further purification.

[1229] MS m / z: 170 [M+l]+

[1230] [Preparative example C-881 1 -(3-aminopropyl)-4-hydroxy- 1 H-pyrazole-5 -carboxylic acid (Int. C-88)

[1231] Int. C-88

[1232] The title compound was synthesized in the same manner as described in preparative example C-87 using Int. C-10.

[1233] MS m / z: 186 [M+l]+

[1234] [Preparative example C-891 (l-(3-aminopropyl)-lH-pyrazol-4-yl)methanol (Int. C-89)

[1235] -128-

[1236] FH13103096.12 Attorney Docket No.: LCH-03225

[1237] (Step 1) benzyl (3-(4-(hydroxymethyl)- IH-pyrazol- 1 -yl)propyl)carbamate

[1238] To a solution of ethyl l-(3-(((benzyloxy)carbonyl)amino)propyl)-lH-pyrazole-4- carboxylate prepared in preparative example C-87 (step 1) (85.9 mg, 0.26 mmol) in THF (2.5 mL) was added LiBF (16.9 mg, 0.78 mmol). The reaction mixture was stirred at 60 °C for 6 h under nitrogen atmosphere. After being cooled to RT, the residue was diluted with distilled water (20 mL), then extracted with EA (20 mL). The organic layer was dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 10) to obtain the title compound as a colorless oil (26.0 mg).

[1239] MS m / z: 290 [M+l]+

[1240] (Step 2) (l-(3-aminopropyl)-lH-pyrazol-4-yl)methanol (Int. C-89)

[1241] The title compound (12.0 mg) was synthesized following the procedure described in preparative example C-87 (step 3) using the compound (16.7 mg, 0.06 mmol) obtained in (step 1) above.

[1242] MS m / z: 156 [M+l]+

[1243] [Preparative example C-901 3-(4-methoxy-lH-pyrazol-l-yl)propan-l-amine (Int. C-90)

[1244] (Step 1) benzyl (3-(4-hydroxy-lH-pyrazol-l-yl)propyl)carbamate

[1245] To amixture of Int. C-4 (749 mg, 3.78 mmol) and benzyl N-(3-bromopropyl)carbamate (1.13 g, 4.15 mmol) dissolved in DMF (12.6 mL) was added K2CO3 (1.85 g, 5.66 mmol). The reaction mixture was stirred at 70 °C for 22 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (100 mL), then extracted three times with EA (100 mL). The combined organic layers were washed with distilled water, dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was dissolved in THF (38 mL), then 1 M TBAF solution in THF (5.7 mL, 5.70 mmol) was added thereto at 0 °C. The mixture was stirred at RT for 1 h under nitrogen atmosphere. The residue was diluted with distilled water (50 mL), then extracted three times with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under

[1246] -129-

[1247] FH13103096.12 Attorney Docket No.: LCH-03225 reduced pressure. The remaining residue was purified by MPLC (MeOH : DCM = 1 : 99 to 5 : 95) to obtain the title compound as a yellow oil (615 mg, 59%).

[1248] ‘ H NMR (400 MHz, CDCh) 5 7.38 - 7.30 (m, 5H), 7.16 (s, 1H), 7.06 (s, 1H), 5.13 (s, 1H), 5.08 (s, 2H), 4.04 (t, J = 6.5 Hz, 2H), 3.13 (q, J = 6.5 Hz, 2H), 1.96 (p, J = 6.4 Hz, 2H).

[1249] (Step 2) benzyl (3-(4-methoxy-lH-pyrazol-l-yl)propyl)carbamate

[1250] To solution of the compound prepared in (step 1) above (50.0 mg, 0.18 mmol) in DMF (1.8 mL), were added CS2CO3 (71.0 mg, 0.22 mmol) and iodomethane (11.3 pL, 0.182 mmol) subsequently. The mixture was stirred at RT for 18 h under nitrogen atmosphere. On completion, the residue was diluted with distilled water (20 mL), then extracted three times with DCM (20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (MeOH : DCM = 5 : 95) to obtain the title compound as a yellow oil (48.9 mg, 93%).

[1251] 1H NMR (400 MHz, CDCh) 5 7.40 - 7.29 (m, 5H), 7.21 (s, 1H), 7.08 (s, 1H), 5.09 (s, 2H), 5.05 (s, 1H), 4.09 (t, J = 6.5 Hz, 2H), 3.72 (s, 3H), 3.16 (q, J = 6.4 Hz, 2H), 2.00 (p, J = 6.4 Hz, 2H).

[1252] (Step 3) 3-(4-methoxy-lH-pyrazol-l-yl)propan-l-amine (Int. C-90)

[1253] The title compound (27.7 mg) was synthesized following the procedure described in preparative example C-87 (step 3) using the compound (48.9 mg, 0.17 mmol) obtained in (step 2) above.

[1254] ’ H NMR (400 MHz, CDCh) 5 7.19 (s, 1H), 7.06 (s, 1H), 4.09 (t, J = 6.8 Hz, 2H), 3.71 (s, 3H), 2.67 (t, J = 6.8 Hz, 2H), 1.93 (p, J = 6.8 Hz, 2H).

[1255] (Preparative example C-911 l-(3-aminopropyl)-4-hydroxypiperidin-2-one (Int. C-91)

[1256] (Step 1) 4-ethoxv-5.6-dihvdropyridin-2(lH)-one

[1257] A mixture of 2,4-piperidinedione (1.00 g, 8.84 mmol) and p-toluenesulfonic acid (152 mg, 0.88 mmol) dissolved in EtOH (18 mL) was stirred at 80 °C for 3 h under nitrogen atmosphere. After being cooled to RT, the solvent was removed under reduced pressure. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined

[1258] -130-

[1259] FH13103096.12 Attorney Docket No.: LCH-03225 organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (MeOH : DCM = 1 : 19) to obtain the title compound as a yellow solid (220 mg, 18%).

[1260] MS m / z: 142 [M+l]+.1H NMR (400 MHz, CDCh) 5 5.29 (s, 1H), 5.04 (s, 1H), 3.93 - 3.87 (m, 2H), 3.44 - 3.40 (m, 2H), 2.45 (t, J = 7.1 Hz, 2H), 1.36 (t, J = 7.1 Hz, 3H).

[1261] (Step 2) benzyl (3-(4-ethoxy-6-oxo-3,6-dihydropyridin-l(2H)-yl)propyl)carbamate

[1262] The title compound (150 mg, 32%) was synthesized following the procedure described in preparative example C-35 (step 1) using the compound (200 mg, 1.42 mmol) obtained in (step 1) above.

[1263] MS m / z: 333 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.40 - 7.29 (m, 5H), 5.98 (t, . / = 6.4 Hz, 1H), 5.09 (s, 2H), 5.04 (s, 1H), 3.89 - 3.84 (m, 2H), 3.49 - 3.44 (m, 2H), 3.35 (t, J = 7.2 Hz, 2H), 3.16 (q, J = 6.4 Hz, 2H), 2.44 (t, J = 7.1 Hz, 2H), 1.74 - 1.61 (m, 2H), 1.34 (t, J = 7.0 Hz, 3H).

[1264] (Step 3) benzyl (3-(2,4-dioxopiperidin-l-yl)propyl)carbamate

[1265] To a solution of the compound prepared in (step 2) above (150 mg, 0.45 mmol) in ACN (1.50 mL) was added 1 N HC1 aqueous solution (3 mL). After stirring at 80 °C for 5 h under nitrogen atmosphere, the reaction was quenched by adding saturated NaHCCh aqueous solution (20 mL), then extracted three times with EA (10 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The title compound was obtained as a light brown oil (70.0 mg), which was used for the next step without further purification.

[1266] MS m / z: 305 [M+l]+

[1267] (Step 4) benzyl (3-(4-hydroxy-2-oxopiperidin-l-yl)propyl)carbamate

[1268] To a solution of the compound prepared in (step 3) above (82.5 mg, 0.27 mmol) in DCM (1.8 mL) were added acetic acid (0.2 mL) and NaBH4 (20.5 mg, 0.54 mmol) at 0 °C. After stirring at RT for 1 h under nitrogen atmosphere, the reaction was quenched by adding distilled water (30 mL) carefully, then extracted twice with DCM (30 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the title compound as a colorless oil (62.0 mg).

[1269] MS m / z: 307 [M+l]+

[1270] (Step 5) l-(3-aminopropyl)-4-hydroxypiperidin-2-one (Int. C-91)

[1271] The title compound (26 mg) was synthesized following the procedure described in

[1272] -131-

[1273] FH13103096.12 Attorney Docket No.: LCH-03225 preparative example C-29 using the compound (62.0 mg, 0.20 mmol) obtained in (step 4).

[1274] MS m / z: 173 [M+l]+

[1275] (Preparative example C-921 l-(3-aminopropyl)-3,4-dihydroxypiperidin-2-one (Int. C-92)

[1276] Int. C-92

[1277] (Step 1) tert-butyl 2, 4-di oxopiperidine- 1 -carboxylate

[1278] To a solution of 2, 2-dimethyl-l,3-dioxane-4, 6-dione (1.67 g, 11.6 mmol) in DCM (35 mL) were added DMAP (1.94 g, 15.9 mmol), 3-((tert-butoxycarbonyl)amino)propanoic acid (2.00 g, 10.6 mmol) and EDC hydrochloride (2.43 g, 12.7 mmol) at 0 °C. The reaction mixture was stirred at RT for 15 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (50 mL) and 5% KHSO4 aqueous solution (10 mL), then extracted twice with DCM (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was dissolved in EA (100 mL), and the solution was stirred at 80 °C for 17 h under nitrogen atmosphere. On completion, the solution was cooled 0 °C, then Et20 and n-hexane were added thereto to afford precipitate, which was filtered, washed with distilled water, then dried in vacuo to give the title compound as a white solid (413 mg, 18%), which was used for the next step without further purification.

[1279] MS m / z: 214 [M+l]+.1H NMR (400 MHz, CDCh) 54.10 (t, J = 6.1 Hz, 2H), 3.51 (s, 2H), 2.63 (t, J = 6.1 Hz, 2H), 1.56 (s, 9H).

[1280] (Step 2) tert-butyl 4-hydroxy-2-oxopiperidine-l -carboxylate

[1281] To a solution of the compound prepared in (step 1) above (100 mg, 0.47 mmol) in DCM (4 mL) were added acetic acid (0.24 mL) and NaBHtCN (35.4 mg, 0.56 mmol) at 0 °C. After stirring at RT for 3 h under nitrogen atmosphere, the reaction was quenched by adding saturated NaHCCL aqueous solution (10 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The title compound was obtained as a colorless oil (100 mg) and used for the next step without further purification.

[1282] MS m / z: 216 [M+l]+

[1283] -132-

[1284] FH13103096.12 Attorney Docket No.: LCH-03225

[1285] (Step 3) tert-butyl 6-oxo-3,6-dihydropyridine-l(2H)-carboxylate

[1286] To a solution of the compound prepared in (step 2) above (200 mg, 0.93 mmol) in DCM (5 mL) was added TEA (392 pL, 2.79 mmol) at 0 °C. After stirring at the same temperature for 0.5 h, acetic anhydride (105 pL, 1.12 mmol) was added thereto, then stirred at RT for 18 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (50 mL) and saturated NaHCOt aqueous solution (10 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n- hexane = 1 : 4) to obtain the title compound as a colorless oil (70.0 mg, 38%).

[1287] MS m / z: 198 [M+l]+.1H NMR (400 MHz, CDCh) 5 6.80 - 6.75 (m, 1H), 5.98 - 5.94 (m, 1H), 3.86 (t, J = 6.5 Hz, 2H), 2.43 - 2.38 (m, 2H), 1.55 (s, 9H).

[1288] (Step 4) 5 ,6-dihydropyridin-2( 1 H)-one

[1289] To a solution of the compound prepared in (step 3) above (75.0 mg, 0.38 mmol) in DCM (1 mL) was added TFA (0.5 mL). The mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The title compound was obtained as a colorless oil (31.4 mg, 85%), which was used for the next step without further purification.

[1290] MS m / z: 98 [M+l]+.1H NMR (400 MHz, CDCh) 5 6.71 - 6.61 (m, 1H), 5.96 - 5.88 (m, 1H), 5.45 (s, 1H), 3.50 - 3.39 (m, 2H), 2.42 - 2.31 (m, 2H).

[1291] (Step 5) benzyl (3-(6-oxo-3,6-dihydropyridin-l(2H)-yl)propyl)carbamate

[1292] The title compound (17.3 mg, 19%) was synthesized following the procedure described in preparative example C-35 (step 1) using the compound (31.4 mg, 0.32 mmol) obtained in (step 4).

[1293] MS m / z: 289 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.36 - 7.30 (m, 5H), 6.59 - 6.48 (m, 1H), 5.94 - 5.88 (m, 1H), 5.83 (s, 1H), 5.09 (s, 2H), 3.50 - 3.42 (m, 2H), 3.37 (t, J = 7.2 Hz, 2H), 3.17 (q, J = 6.3 Hz, 2H), 2.44 - 2.30 (m, 2H), 1.76 - 1.65 (m, 2H).

[1294] (Step 6) benzyl (3-(3,4-dihydroxy-2-oxopiperidin- l-yl)propyl)carbamate

[1295] To a solution of the compound prepared in (step 5) above (17.3 mg, 0.06 mmol) in t- BuOH (1.5 mL) were added 4-methylmorpholine N-oxide (28.1 mg, 0.24 mmol) and 2.5 wt.% OsO4solution in t-BuOH (75.2 pL, 0.01 mmol). The reaction mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered,

[1296] -133-

[1297] FH13103096.12 Attorney Docket No.: LCH-03225 then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 20) to obtain the title compound as a colorless oil (9.0 mg, 47%).

[1298] MS m / z: 323 [M+l]+.1H NMR (400 MHz, CDCh) 57.38 - 7.28 (m, 5H), 5.65 (s, 1H), 5.10 (s, 2H), 4.29 (d, J = 4.4 Hz, 1H), 3.99 (d, J = 3.0 Hz, 1H), 3.93 (s, 1H), 3.68 - 3.55 (m, 2H), 3.33 - 3.20 (m, 2H), 3.19 - 3.02 (m, 2H), 2.71 (s, 1H), 2.18 - 2.08 (m, 1H), 2.05 - 1.94 (m, 1H), 1.81 - 1.70 (m, 2H).

[1299] (Step 7) l-(3-aminopropyl)-3,4-dihydroxypiperidin-2-one (Int. C-92)

[1300] The title compound (5.20 mg) was synthesized following the procedure described in preparative example C-29 using the compound (9.00 mg, 0.03 mmol) obtained in (step 6).

[1301] MS m / z: 189 [M+l]+

[1302] [Preparative example C-931 5-amino-2-(3-aminopropyl)pyridazin-3(2H)-one (Int. C-93)

[1303] (Step 1) benzyl (3-(4,5-dichloro-6-oxopyridazin- l(6H)-yl)propyl)carbamate

[1304] The title compound (190 mg, 88%) was synthesized following the procedure described in preparative example C-35 (step 1) using 4,5-dichloropyridazin-3(2H)-one (100 mg, 0.60 mmol).

[1305] MS m / z: 357 [M+l]+.1H NMR (400 MHz, CDCh) 57.80 (s, 1H), 7.37 - 7.32 (m, 5H), 5.31 (s, 1H), 5.09 (d, J = 8.3 Hz, 2H), 4.28 - 4.26 (m, 2H), 3.22 - 3.18(m, 2H), 2.03 - 1.98(m, 2H).

[1306] (Step 2) benzyl (3 -(4-amino-5 -chloro-6-oxopyridazin- 1 (6H)-yl)propyl)carbamate

[1307] The title compound (31.0 mg, 36%) was synthesized following the procedure described in preparative example C-44 (step 3) using the compound (90.0 mg, 0.25 mmol) obtained in (step 1) above.

[1308] MS m / z: 337 [M+l]+

[1309] (Step 3) 5-amino-2-(3-aminopropvl)pvridazin-3(2H)-one (Int. C-93)

[1310] The title compound (15.0 mg, 95%) was synthesized following the procedure described in preparative example C-29 using the compound (31.5 mg, 0.09 mmol) obtained in (step 2) above.

[1311] MS m / z: 169 [M+l]+

[1312] [Preparative example C-94] 6-(3-aminopropyl)pyridin-2-amine (Int. C-94)

[1313] -134-

[1314] FH13103096.12 Attorney Docket No.: LCH-03225

[1315] (Step 1) benzyl prop-2-yn-l-ylcarbamate

[1316] To a solution of propargylamine (0.500 mL, 7.87 mmol) in DCM (20 mL) were added TEA (1.66 mL, 11.8 mmol) and benzyl chloroformate (1.29 mL, 8.66 mmol) at 0 °C. After stirring at RT for 3 h under nitrogen atmosphere, the reaction was quenched by adding distilled water (100 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 1 : 9 to 2 : 8) to obtain the title compound as a colorless oil (984 mg, 66%).

[1317] MS m / z: 212 [M+23]+. ’ H NMR (400 MHz, CDCh) 57.35 - 7.32 (m, 5H), 5.13 (s, 2H), 4.00 (s, 2H), 2.26 - 2.20 (m, 1H).

[1318] (Step 2) benzyl (3-(6-aminopyridin-2-yl)prop-2-yn-l-yl)carbamate

[1319] To a solution of the compound prepared in (step 1) above (393 mg, 2.08 mmol) in DMF (4 mL) were added 2-amino-6-bromopyridine (300 mg, 1.73 mmol), TEA (0.487 mL, 3.47 mmol), Cui (33.0 mg, 0.17 mmol), and PdChfPPlb (121 mg, 0.17 mmol). The reaction mixture was stirred at 100 °C for 16 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 1 : 1 to 3 : 1) to obtain the title compound as a brown oil (201 mg, 41%).

[1320] MS m / z: 282 [M+l]+

[1321] (Step 3) 6-(3-aminopropyl)pyridin-2-amine (Int. C-94)

[1322] The title compound (71.0 mg) was synthesized following the procedure described in preparative example C-29 using the compound (201 mg, 0.72 mmol) obtained in (step 2) above.

[1323] MS m / z: 152 [M+l]+

[1324] [Preparative example C-951 3-(4-aminobutyl)pyridin-2-amine (Int. C-95)

[1325] -135-

[1326] FH13103096.12 Attorney Docket No.: LCH-03225

[1327] (Step 1) 3-bromo-5-chloropyridin-2-amine

[1328] To a solution of 5-chloropyridin-2-amine (300 mg, 2.33 mmol) in chloroform (6 mL) was added bromine (0.12 mL, 2.33 mmol), then the reaction mixture was stirred at RT for 2 h under nitrogen atmosphere. The reaction was quenched by adding saturated NaHCCh aqueous solution (50 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 5) to obtain the title compound as a light brown solid (373 mg, 77%).

[1329] MS m / z: 208 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.98 (d, J = 2.3 Hz, 1H), 7.66 (d, J = 2.3 Hz, 1H), 4.90 (s, 2H).

[1330] (Step 2) tert-butyl (4-(2-amino-5-chloropyridin-3-yl)but-3-yn-l-yl)carbamate

[1331] The title compound (43.1 mg) was synthesized following the procedure described in preparative example C-94 (step 2) using the compound (100 mg, 0.48 mmol) obtained in (step 1) above and tert-butyl N-but-3-ynylcarbamate (89.7 mg, 0.53 mmol).

[1332] MS m / z: 296 [M+l]+.1H NMR (400 MHz, CDCh) 57.93 (s, 1H), 7.42 (s, 1H), 5.09 (s, 2H), 4.88 (s, 1H), 3.38 (d, J = 6.5 Hz, 2H), 2.64 (t, J = 6.4 Hz, 2H), 1.43 (s, 9H).

[1333] (Step 3) tert-butyl (4-(2-aminopyridin-3-yl)butyl)carbamate

[1334] The title compound (27.8 mg) was synthesized following the procedure described in preparative example C-29 using the compound (43.1 mg) obtained in (step 2) above.

[1335] MS m / z: 266 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.87 (dd, J = 5.1, 1.9 Hz, 1H), 7.24 (d, J = 1.9 Hz, 1H), 6.60 (dd, J = 7.3, 5.1 Hz, 1H), 4.94 (s, 2H), 4.61 (s, 1H), 3.19 (d, J = 6.6 Hz, 2H), 2.46 (t, J = 7.8 Hz, 2H), 1.66 - 1.59 (m, 2H), 1.58 - 1.51 (m, 2H), 1.43 (s, 9H).

[1336] (Step 4) 3-(4-aminobutyl)pyridin-2-amine (Int. C-95)

[1337] The title compound (25.0 mg) was synthesized following the procedure described in preparative example C-92 (step 4) using the compound (27.8 mg) obtained in (step 3) above.

[1338] MS m / z: 166 [M+l]+

[1339] [Preparative example C-961 6-(3-aminopropyl)-2-fluoropyridin-3-amine (Int. C-96)

[1340] Int. C-96

[1341] The title compound was synthesized in the same manner as described in preparative example C-95 using 2-fluoropyridin-3-amine and tert-butyl prop-2-yn-l-ylcarbamate.

[1342] -136-

[1343] FH13103096.12 Attorney Docket No.: LCH-03225

[1344] MS m / z: 170 [M+l]+. ’ H NMR (400 MHz, DMSO-76) 5 7.78 (s, 2H), 7.09 (dd, J = 11.3, 7.8 Hz, 1H), 6.89 (dd, 7 = 7.8, 1.8 Hz, 1H), 2.77 (q, 7 = 5.9 Hz, 2H), 2.56 (t, 7 = 7.5 Hz, 2H), 1.89 - 1.79 (m, 2H).

[1345] [Preparative example C-97] 5-(3-aminopropyl)-2-fluoropyridin-3-amine (Int. C-97)

[1346] Int. C-97

[1347] The title compound was synthesized in the same manner as described in preparative example C-95 using 5-bromo-2-fluoropyridin-3-amine and tert-butyl prop-2-yn-l-ylcarbamate, except for the omission of the bromination step.

[1348] MS m / z: 170 [M+l]+. ’ H NMR (400 MHz, DMSO-&) 5 7.84 (s, 2H), 7.17 (t, 7 = 2.3 Hz, 1H), 6.97 (dd, J = 10.7, 2.4 Hz, 1H), 2.76 (q, J = 6.3 Hz, 2H), 2.54 (s, 2H), 1.78 (p, J = 7.6 Hz, 2H).

[1349] [Preparative example C-981 5-(3-aminopropyl)pyridin-3-ol (Int. C-98)

[1350] Int. C-98

[1351] The title compound was synthesized in the same manner as described in preparative example C-95 using Int. C-5 and tert-butyl prop-2-yn-l-ylcarbamate, except for the omission of the bromination step.

[1352] MS m / z: 153 [M+l]+

[1353] [Preparative example C-991 3-(pyrimidin-4-yloxy)propan-l -amine (Int. C-99)

[1354] Int. C-99

[1355] (Step 1) tert-butyl (3-(pyrimidin-4-yloxy)propyl)carbamate

[1356] The title compound (43.4 mg, 60%) was synthesized following the procedure described in preparative example C-36 (step 2) using tert-butyl (3-hydroxypropyl)carbamate (50.0 mg, 0.29 mmol) and 4-hydroxypyrimidine (30.2 mg, 0.31 mmol).

[1357] MS m / z: 254 [M+l]+.1H NMR (400 MHz, CDC13) 58.76 (s, 1H), 8.42 (dd, J = 5.8, 0.6 Hz, 1H), 6.72 (dd, J = 5.8, 1.2 Hz, 1H), 4.74 (s, 1H), 4.44 (t, J = 6.2 Hz, 2H), 3.34 - 3.19 (m, 2H), 1.98 (p, 7 = 6.4 Hz, 2H), 1.44 (s, 9H).

[1358] (Step 2) 3-(pyrimidin-4-yloxy)propan-l-amine (Int. C-99)

[1359] The title compound (39.1 mg) was synthesized following the procedure described in

[1360] -137-

[1361] FH13103096.12 Attorney Docket No.: LCH-03225 preparative example C-92 (step 4) using the compound (43.4 mg, 0.17 mmol) obtained in (step 1) above.

[1362] MS m / z: 154 [M+l]+. ‘ H NMR (400 MHz, DMSO-&) 5 8.85 (s, 1H), 8.57 (d, J = 5.9 Hz, 1H), 7.94 (s, 3H), 7.02 (dd, J = 5.9, 1.3 Hz, 1H), 4.42 (t, J = 6.3 Hz, 2H), 2.99 - 2.88 (m, 2H), 2.10 - 1.98 (m, 2H).

[1363] [Preparative example C-1001 N2-(2-aminoethyl)pyridine-2,6-diamine (Int. C-100)

[1364] (Step 1) tert-butyl (6-bromopyridin-2-yl)(tert-butoxycarbonyl)carbamate

[1365] The title compound (811 mg, 63%) was synthesized following the procedure described in preparative example C-ll (step 4) using 6-bromopyridin-2-amine (600 mg, 3.47 mmol).

[1366] MS m / z: 374 [M+l]+

[1367] (Step 2) tert-butyl (tert-butoxycarbonyl)(6-((2-((tert-butoxycarbonyl)amino)ethyl) amino)pyridin-2-yl)carbamate

[1368] To a solution of the compound prepared in (step 1) above (811 mg, 2.17 mmol) in 1.4- dioxane (10 mL) were added then N-Boc-ethylenediamine (0.344 mL, 2.17 mmol), CS2CO3 (2.12 g, 6.52 mmol), xantphos (126 mg, 0.22 mmol) and Pd2(dba)t (99.5 mg, 0.11 mmol). The reaction mixture was stirred at 95°C for 6 h under nitrogen atmosphere. On completion, the reaction mixture was cooled to RT, diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 1 : 3 to 1 : 1) to obtain the title compound as a yellow oil (741 mg, 75%).

[1369] MS m / z: 453 [M+l]+

[1370] (Step 3) N2-(2-aminoethvl)pyridine-2,6-diamine (Int. C-100)

[1371] The title compound (251 mg) was synthesized following the procedure described in preparative example C-92 (step 4) using the compound (741 mg, 1.64 mmol) obtained in (step 2) above.

[1372] MS m / z: 153 [M+l]+

[1373] [Preparative example C-1011 l-(3-aminopropyl)-lH-pyrazol-4-ol (Int. C-101)

[1374] Int C-101

[1375] (Step 1) tert-butyl (3-(4-((tert-butyldimethylsilyl)oxy)- IH-pyrazol- 1 - yl)propyl)carbamate

[1376] -138-

[1377] FH13103096.12 Attorney Docket No.: LCH-03225

[1378] To a mixture Int. C-4 (332 mg, 1.67 mmol) and K2CO3 (578 mg, 4.18 mmol) dissolved in DMF (6 mL) was added tert-butyl N-(3-bromopropyl) carbamate (438 mg, 1.84 mmol). The reaction mixture was stirred at 60 °C for 6 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (DCM : n-hexane= 1 : 1 to 4 : 1) to obtain the title compound as a brown oil (260 mg, 44%).

[1379] MS m / z: 356 [M+l]+

[1380] (Step 2) l-(3-aminopropyl)-lH-pyrazol-4-ol (Int. C-101)

[1381] The title compound (181 mg) was synthesized following the procedure described in preparative example C-92 (step 4) using the compound (260 mg, 0.73 mmol) obtained in (step 1) above.

[1382] MS m / z: 142 [M+l]+.1H NMR (400 MHz, DMSO-&) 5 8.05 - 7.75 (m, 3H), 7.24 (s, 1H), 7.02 (s, 1H), 4.04 (t, J = 6.6 Hz, 2H), 2.72 - 2.65 (m, 2H), 2.02 - 1.93 (m, 2H). (Preparative example C-102] l-(3-aminopropyl)-lH-pyrazol-3-amine (Int. C-102)

[1383] Int. c-102

[1384] The title compounds synthesized in the same manner as described in preparative example C-101 using 3-amino-lH-pyrazole.

[1385] MS m / z: 141 [M+l]+

[1386] (Preparative example C-1031 3-(lH-pyrazol-5-yl)propan-l-amine (Int. C-103)

[1387] (Step 1) 1 -(4-methoxybenzyl)- IH-pyrazole

[1388] The title compound (951 mg, 43%) was synthesized following the procedure described in preparative example C-18 (step 1) using IH-pyrazole (800 mg, 11.8 mmol).

[1389] MS m / z: 189 [M+l]+.1H NMR (400 MHz, CDCI3) 5 7.53 (d, J = 2.1 Hz, 1H), 7.34 (d, J = 2.4 Hz, 1H), 7.21 - 7.13 (m, 2H), 6.91 - 6.83 (m, 2H), 6.25 (d, J = 2.1 Hz, 1H), 5.25 (s, 2H), 3.78 (s, 3H).

[1390] -139-

[1391] FH13103096.12 Attorney Docket No.: LCH-03225

[1392] (Step 2) 1 -(4- methoxybenzyl)- 1 H-pyrazole-5 -carbaldehyde

[1393] The title compound (268 mg, 52%) was synthesized following the procedure described in preparative example C-8 (step 2) using the compound (450 mg, 2.39 mmol) obtained in (step 1) above.

[1394] MS m / z: 217 [M+l]+.1H NMR (400 MHz, CDCh) 5 9.84 (s, 1H), 7.58 (d, J = 2.3 Hz, 1H), 7.30 - 7.22 (m, 2H), 6.91 (d, J = 2.1 Hz, 1H), 6.83 (d, J = 8.8 Hz, 2H), 5.66 (s, 2H), 3.76 (s, 3H).

[1395] (Step 3) (E)-3-(l-(4-methoxybenzyl)-lH-pyrazol-5-yl)acrylonitrile

[1396] To a solution of cyanomethyl(triphenyl)phosphonium bromide (189 mg, 0.50 mmol) in THF (5 mL) was added NaH (21.0 mg, 0.53 mmol, 60% in dispersion in mineral oil) at 0 °C. After stirring at 0 °C for 15 min, the compound prepared in (step 2) above (100 mg, 0.46 mmol) was added thereto. The reaction mixture was stirred at 60 °C for 0.5 h under nitrogen atmosphere. After being cooled to RT, the reaction was quenched by adding saturated NH4CI aqueous solution (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 1 : 5) to obtain the title compound as a white solid (85.1 mg, 77%).

[1397] MS m / z: 240 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.54 (dd, J = 2.2, 0.7 Hz, 1H), 7.23 (d, J = 16.4 Hz, 1H), 7.05 (d, J = 8.9 Hz, 2H), 6.86 (d, J = 8.9 Hz, 2H), 6.60 (d, J = 2.3 Hz, 1H), 5.73 (d, J = 16.4 Hz, 1H), 5.36 (s, 2H), 3.79 (s, 3H).

[1398] (Step 4) 3-( 1 -(4-methoxybenzyl)- 1 H-pyrazol-5 -yl)propan- 1 -amine

[1399] To a solution of the compound prepared in (step 3) above (85.1 mg, 0.36 mmol) in a mixed solvent of MeOH / EtOH (3 mL, 1 : 1) was added Raney-Ni (170 mg). After stirring at RT for 17 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to obtain the title compound as an off-white solid (87.0 mg), which was used for the next step without further purification.

[1400] MS m / z: 246 [M+l]+

[1401] (Step 5) 3-(lH-pyrazol-5-yl)propan-l-amine (Int. C-103)

[1402] A mixture of the compound prepared in (step 4) above (130 mg, 0.53 mmol) and TFA (3.3 mL) dissolved in DCM (10 mL) was stirred at 80 °C for 12 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure to afford the title compound as a yellow solid (67 mg), which was used for the next step without further purification.

[1403] -140-

[1404] FH13103096.12 Attorney Docket No.: LCH-03225

[1405] MS m / z: 126 [M+l]+

[1406] [Prepa

[1407] (Step 1) N-(benzyloxy)-5 -bromopentanamide

[1408] To a solution of 5-bromopentanoyl chloride (500 mg, 2.51 mmol) in DCM (9 mL) were added O-benzylhydroxylamine hydrochloride (400 mg, 2.51 mmol) and pyridine (406 pL, 5.01 mmol). The reaction mixture was stirred at RT for 4 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with saturated NaHCCh aqueous solution (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a colorless oil (658 mg), which was used for the next step without further purification.

[1409] MS m / z: 287 [M+l]+.1H NMR (400 MHz, CDCh) 57.93 (s, 1H), 7.40 - 7.37 (m, 5H), 4.92 (s, 2H), 3.39 (t, J = 6.5 Hz, 2H), 2.17 - 1.99 (m, 2H), 1.90 - 1.83 (m, 2H), 1.82 - 1.74 (m, 2H).

[1410] (Step 2) 1 -(benzyloxy )piperidin-2-one

[1411] To a solution of the compound prepared in (step 1) above (717 mg, 2.51 mmol) in DCM (11 mL) were added benzyltriethylammonium chloride (57.1 mg, 0.25 mmol) and 5% NaOH aqueous solution (2.80 mL). After stirring at RT for 16 h under nitrogen atmosphere, the reaction was quenched by adding 1 N HC1 aqueous solution (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 3 : 2) to obtain the title compound as a colorless oil (376 mg, 73% for 2 steps).

[1412] MS m / z: 206 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.46 - 7.41 (m, 2H), 7.39 - 7.34 (m, 3H), 4.98 (s, 2H), 3.34 (t, J = 6.1 Hz, 2H), 2.45 (t, J = 6.4 Hz, 2H), 1.82 - 1.74 (m, 2H), 1.74 - 1.68 (m, 2H).

[1413] FH13103096.12 Attorney Docket No.: LCH-03225

[1414] (Step 3) 3 -allyl- 1 -(benzyl oxy )piperidin-2-one

[1415] To a solution of the compound prepared in (step 2) above (376 mg, 1.83 mmol) in THF (20 mL) were added 1 M LiHMDS solution in THF (2 mL, 2.02 mmol) and HMPA (956 pL, 5.50 mmol) at -70 °C. After stirring at the same temperature for 0.5 h, 3-bromo-l -propene (167 pL, 1.92 mmol) was added. The mixture was stirred at RT for 19 h under nitrogen atmosphere. On completion, the reaction was quenched by adding distilled water (100 mL), diluted with saturated CuSO4 aqueous solution (10 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 3) to obtain the title compound as a colorless oil (228 mg, 51%).

[1416] MS m / z: 246 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.45 - 7.40 (m, 2H), 7.37 - 7.31 (m, 3H), 5.83 - 5.70 (m, 1H), 5.10 - 5.01 (m, 2H), 4.98 - 4.90 (m, 2H), 3.39 - 3.23 (m, 2H), 2.72 - 2.62 (m, 1H), 2.47 - 2.38 (m, 1H), 2.31 - 2.20 (m, 1H), 1.86 - 1.77 (m, 2H), 1.72 - 1.62 (m, 1H), 1.51 - 1.38 (m, 1H).

[1417] (Step 4) 1 -(benzyloxy)-3-(3-hydroxypropyl)piperidin-2-one

[1418] To BH3-SMe2 complex solution in THF (0.18 mL, 1.94 mmol) diluted in THF (3 mL) was added cyclohexene (393 pL, 3.88 mmol) at 0 °C. The reaction mixture was stirred at the same temperature for 0.5 h. The compound prepared in (step 3) above (317 mg, 1.29 mmol) was added thereto, and the reaction mixture was stirred at the same temperature for 1 h under nitrogen atmosphere. To the reaction mixture were added 10% NaOH aqueous solution (3 mL) and 30% hydrogen peroxide aqueous solution (3 mL), then the reaction mixture was allowed to stir at RT for 19 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 25) to obtain the title compound as a colorless oil (149 mg, 44%).

[1419] MS m / z: 264 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.43 - 7.37 (m, 2H), 7.36 - 7.31 (m, 3H), 4.97 - 4.87 (m, 2H), 3.63 (t, J = 6.1 Hz, 2H), 3.36 - 3.28 (m, 2H), 2.90 (s, 1H), 2.44 - 2.35 (m, 1H), 1.97 - 1.87 (m, 1H), 1.87 - 1.78 (m, 2H), 1.74 - 1.62 (m, 2H), 1.61 - 1.54 (m, 2H), 1.50 - 1.40 (m, 1H).

[1420] (Step 5) 3-(l-(benzyloxy)-2-oxopiperidin-3-yl)propyl methanesulfonate

[1421] To a solution of the compound prepared in (step 4) above (149 mg, 0.57 mmol) in DCM (5 mL) were added TEA (237 pL, 1.70 mmol) and methanesulfonyl chloride (65.7 pL, 0.85

[1422] -142-

[1423] FH13103096.12 Attorney Docket No.: LCH-03225 mmol) at 0 °C. The mixture was stirred at RT for 1 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (50 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a light orange oil (219 mg), which was used for the next step without further purification.

[1424] MS m / z: 342 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.45 - 7.40 (m, 2H), 7.39 - 7.34 (m, 3H), 5.00 - 4.90 (m, 2H), 4.31 - 4.21 (m, 2H), 3.41 - 3.33 (m, 2H), 3.02 (s, 3H), 2.45 - 2.36 (m, 1H), 2.01 - 1.92 (m, 1H), 1.91 - 1.82 (m, 4H), 1.79 - 1.70 (m, 1H), 1.67 - 1.59 (m, 1H), 1.53 - 1.44 (m, 1H).

[1425] (Step 6) 3-(3-azidopropyl)- 1 -(benzyloxy)piperidin-2-one

[1426] A mixture of the compound prepared in (step 5) above (112 mg, 0.33 mmol) and Nabh (42.7 mg, 0.66 mmol) dissolved in DMF (1.5 mL) was stirred at RT for 18 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (50 mL), then extracted twice with EA (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 2) to obtain the title compound as a colorless oil (51.2 mg, 54%).

[1427] MS m / z: 289 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.45 - 7.40 (m, 2H), 7.39 - 7.33 (m, 3H), 4.99 - 4.90 (m, 2H), 3.39 - 3.27 (m, 4H), 2.43 - 2.35 (m, 1H), 2.00 - 1.83 (m, 3H), 1.76 - 1.64 (m, 3H), 1.61 - 1.51 (m, 1H), 1.50 - 1.41 (m, 1H).

[1428] (Step 7) 3-(3-aminopropyl)piperidin-2-one (Int. C-104)

[1429] The title compound (29.8 mg) was synthesized following the procedure described in preparative example C-29 using the compound (53.1 mg, 0.18 mmol) obtained in (step 6).

[1430] MS m / z: 157 [M+l]+

[1431] [Preparative example C-1051 l-(3-aminopropyl)-N,N-dimethylpyrrolidin-3-amine (Int. C- 105)

[1432] (Step 1) 2-(3-(3-aminopvrrolidin-l-vl)propyl)isoindoline-l, 3-dione

[1433] To a solution of the compound (85.1 mg, 0.23 mmol) which was prepared in preparative

[1434] -143-

[1435] FH13103096.12 Attorney Docket No.: LCH-03225 example C-13 (step 1) in DCM (2 mL) was added 4 N HC1 solution in 1,4-dioxane (1 mL). The reaction mixture was stirred at RT for 16 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The title compound was obtained as a white solid (81.1 mg), which was used for the next step without further purification.

[1436] MS m / z: 274 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 10.62 (s, 1H), 8.31 (s, 2H), 7.87 (s, 3H), 4.04 - 3.80 (m, 2H), 3.70 - 3.45 (m, 5H), 3.16 - 2.99 (m, 2H), 2.28 - 1.90 (m, 4H).

[1437] (Step 2) 2-(3-(3-(dimethvlamino)pvrrolidin-l-vl)propyl)isoindoline-l.3-dione

[1438] To a solution of the compound prepared in (step 1) above (61.0 mg, 0.197 mmol) in MeOH (3 mL) were added NaBH(OAc)s (125 mg, 0.59 mmol) and 37 wt% formaldehyde solution (22.0 pL, 0.79 mmol). The reaction mixture was stirred at 60 °C for 2 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (30 mL) carefully, then extracted twice with DCM (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 5) to obtain the title compound as a colorless oil (45.0 mg, 57%).

[1439] MS m / z: 302 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.85 - 7.80 (m, 2H), 7.72 - 7.67 (m, 2H), 3.78 - 3.72 (m, 2H), 2.82 - 2.64 (m, 3H), 2.58 - 2.50 (m, 1H), 2.50 - 2.41 (m, 2H), 2.36 - 2.28 (m, 1H), 2.21 (s, 6H), 1.91 - 1.81 (m, 3H), 1.73 - 1.63 (m, 1H).

[1440] (Step 3) l-(3-aminopropyl)-N,N-dimethylpyrrolidin-3-amine (Int. C-105)

[1441] The title compound (24.7 mg, 96%) was synthesized following the procedure described in preparative example C-18 (step 2) using the compound (45.0 mg, 0.15 mmol) obtained in (step 2) above.

[1442] MS m / z: 172 [M+l]+.1H NMR (400 MHz, CDCh) 5 2.89 - 2.71 (m, 5H), 2.58 - 2.32 (m, 4H), 2.21 (s, 6H), 2.05 - 1.93 (m, 2H), 1.70 - 1.63 (m, 2H).

[1443] [Preparative example C-106] l-(3-amino-2-hydroxypropyl)-lH-pyrazol-4-ol (Int. C-106)

[1444] Int. C-106

[1445] (Step 1) benzyl allylcarbamate

[1446] To a solution of allylamine (570 mg, 9.98 mmol) in a mixed solvent of EA / distilled water (30 mL, 1 : 1) were added K2CO3 (3.44 g, 25.0 mmol) and benzyl chloroformate (1.49 mL, 10.5 mmol) at 0 °C. After stirring at RT for 2 h under nitrogen atmosphere, the reaction -144-

[1447] FH13103096.12 Attorney Docket No.: LCH-03225 was quenched by adding 1 N HC1 aqueous solution (50 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a colorless oil (1.90 g, 99%), which was used for the next step without further purification.

[1448] MS m / z: 192 [M+l]+. ’ H NMR (400 MHz, CDC13) 5 7.43 - 7.28 (m, 5H), 5.90-5.80 (m, 1H), 5.22-5.15 (m, 2H), 5.12 (s, 2H), 4.81 (s, 1H), 3.83 (t, J = 5.9 Hz, 2H).

[1449] (Step 2) benzyl (oxiran-2-ylmethyl)carbamate

[1450] To a solution of the compound prepared in (step 1) above (1.90 g, 9.93 mmol) in DCM (25 mL) was added 3-chloroperoxybenzoic acid (3.40 g, 19.8 mmol). The reaction mixture was stirred at RT for 16 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (100 mL) and 1 N NaOH aqueous solution (30 mL), then extracted twice with DCM (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n- hexane = 0 : 10 to 1 : 1) to obtain the mixture compound as a colorless oil (1.80 g, 87%).

[1451] MS m / z: 208 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.36 (d, J = 5.4 Hz, 5H), 5.11 (s, 2H), 4.95 (s, 1H), 3.66-3.60 (m, 1H), 3.30-3.25 (m, 1H), 3.13-3.09 (m, 1H), 2.79 (t, J = 4.4 Hz, 1H), 2.61-2.59 (m, 1H).

[1452] (Step 3) benzyl (2-hydroxy-3-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrazol- 1 -yl)propyl)carbamate

[1453] To a mixture of pyrazole-4-boronic acid pinacol ester (3.6 g, 18.5 mmol) and K2CO3 (3.84 g, 27.8 mmol) dissolved in DMF (36 mL) was added the compound prepared in (step 2) above (5.76 g, 27.8 mmol). The reaction mixture was stirred at 60 °C for 16 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The residue was diluted with distilled water (100 mL), then extracted twice with EA (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (EA : n-hexane = 0 : 10 to 10 : 0) to obtain the title compound as a white solid (2.0 g, TYo).

[1454] MS m / z: 402 [M+l]+’ H NMR (400 MHz, CDC13) 5 7.79 (s, 1H), 7.70 (s, 1H), 7.37- 7.32 (m, 5H), 5.29 (s, 1H), 5.11 (s, 2H), 4.24 - 4.04 (m, 4H), 3.41-3.37 (m, 1H), 3.1-3.13 (m, 1H), 1.31 (s, 12H).

[1455] (Step 4) benzyl (2-hydroxy-3-(4-hydroxy- 1 H-pyrazol- 1 -yl)propyl)carbamate

[1456] To a solution of the compound prepared in (step 3) above (200 mg, 0.50 mmol) in THF (5 mL) were added 2 N NaOH aqueous solution (0.5 mL, 1.00 mmol) and 30% hydrogen

[1457] -145-

[1458] FH13103096.12 Attorney Docket No.: LCH-03225 peroxide solution (0.1 mL, 1.00 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h. On completion, the reaction mixture was quenched by adding 1 N HC1 aqueous solution (50 mL), then extracted twice with 10% MeOH / DCM (50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a colorless oil (102 mg, 70%), which was used for the next step without further purification.

[1459] MS m / z: 292 [M+l]+

[1460] (Step 5) l-(3-amino-2-hydroxypropyl)-lH-pyrazol-4-ol (Int. C-106)

[1461] The title compound (46 mg, 84%) was synthesized following the procedure described in preparative example C-29 using the compound (102 mg, 0.35 mmol) obtained in (step 4).

[1462] MS m / z: 158 [M+l]+

[1463] [Preparative example C-1071 tert-butyl (l-(3-(((benzyloxy)carbonyl)amino)-2- (hydroxymethyl)propyl)-lH-pyrazol-4-yl)carbamate (Int. C-107)

[1464] Int. C-107

[1465] (Step 1) tert-butyl ( 1 -(2-cyano-3-hydroxypropyl)- 1 H-pyrazol-4-yl)carbamate

[1466] To a solution of Int. C-3 (500 mg, 2.73 mmol) in ACN (9 mL) were added 2-(hydroxy methyl)prop-2-enenitrile (0.33 mL, 4.11 mmol) and 1 ,8-diazabicyclo [5.4.0]-7-undecene (0.21 mL, 1.41 mmol). The reaction mixture was stirred at RT for 16 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 3 : 7 to 7 : 3) to obtain the title compound as a white solid (582 mg, 80%).

[1467] MS m / z: 267 [M+l]+.1H NMR (400 MHz, CDCh) 57.76 (s, 1H), 7.43 (s, 1H), 6.26 (s, 1H), 4.73 - 4.29 (m, 2H), 3.71 (d, J = 5.8 Hz, 2H), 3.40 - 3.32 (m, 1H), 3.28 - 3.11 (m, 1H), 1.50 (s, 9H).

[1468] (Step 2) tert-butyl (l-(3-(((benzyloxy)carbonyl)amino)-2-(hydroxymethyl)propyl)-lH- pyrazol-4-yl)carbamate (Int. C-107)

[1469] To a solution of the compound prepared in (step 1) above (582 mg, 2.19 mmol) in a mixed solvent of MeOH / THF (20 mL, 3 : 1) were added Raney-Ni (642 mg) and 28-30% ammonia aqueous solution (1.02 mL). After stirring at RT for 22 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure. The remaining residue (591 mg) was dissolved in THF (15 mL), then

[1470] -146-

[1471] FH13103096.12 Attorney Docket No.: LCH-03225

[1472] NaHCO t (367 mg, 4.37 mmol) in water (5 mL) and benzyl chloroformate (0.32 mL, 2.24 mmol) were added thereto. The reaction mixture was stirred at RT for 3 h under nitrogen atmosphere. On completion, the reaction was quenched by adding distilled water (20 mL), then extracted three times with DCM (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (DCM: MeOH = 100 : 0 to 95 : 5) to obtain the title compound as a white solid (737 mg, 83%).

[1473] MS m / z: 405 [M+l]+.1H NMR (400 MHz, CDCh) 57.64 (s, 1H), 7.35 - 7.32 (m, 5H), 7.26 (s, 1H), 6.30 (s, 1H), 5.36 (s, 1H), 5.11 (s, 2H), 4.27 - 3.90 (m, 2H), 3.56 (s, 1H), 3.66 - 3.34 (m, 2H), 3.16 (ddt, J = 75.6, 14.1, 6.6 Hz, 2H), 2.36 - 2.19 (m, 1H), 1.49 (s, 9H).

[1474] [Preparative example C-1081 tert-butyl (l-(3-amino-2-

[1475] ((dimethylamino)methyl)propyl)-lH-pyrazol-4-yl)carbamate (Int. C-108)

[1476] (Step 1) 3-(((benzyloxy)carbonyl)amino)-2-((4-((tert-butoxycarbonyl)amino)-lH- pyrazol- 1 -yl)methyl)propyl methanesulfonate

[1477] To a solution of the Int. C-107 (737 mg, 1.82 mmol) in DCM (18 mL) were added TEA (0.38 mL, 2.70 mmol) and methanesulfonyl chloride (0.14 mL, 1.81 mmol) at 0 °C. The mixture was stirred at RT for 18 h under nitrogen atmosphere. The reaction was quenched by adding distilled water (10 mL), then extracted twice with DCM (40 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (DCM : MeOH = 97 : 3) to obtain the title compound as a white solid (348 mg, 40%).

[1478] MS m / z: 483 [M+l]+.1H NMR (400 MHz, CDCh) 57.65 (s, 1H), 7.36 (s, 6H), 6.26 (s, 1H), 5.38 (s, 1H), 5.10 (s, 2H), 4.21 - 4.07 (m, 4H), 3.23 (tq, J = 14.0, 7.3 Hz, 2H), 3.04 (s, 3H), 2.58 - 2.50 (m, 1H), 1.49 (s, 9H).

[1479] (Step 2) tert-butyl (1 -(3 -(((benzyl oxy)carbonyl)amino)-2-

[1480] ((dimethylamino)methyl)propyl) - 1 H-pyrazol-4-yl)carbamate

[1481] To a solution of the compound prepared in (step 1) above (348 mg, 0.72 mmol) in THF (4 mL) was added 2 M dimethylamine solution in THF (1.80 mL, 3.61 mmol). The reaction mixture was stirred at 60 °C for 18 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure to obtain the title compound as a white solid (101 mg, 33%), which was used for the next step without further purification.

[1482] -147-

[1483] FH13103096.12 Attorney Docket No.: LCH-03225

[1484] MS m / z: 432 [M+l]+

[1485] (Step 3) tert-butyl (l-(3-amino-2-((dimethylamino)methyl)propyl)-lH-pyrazol-4-yl) carbamate (Int. C-108)

[1486] The title compound (71.2 mg, 99%) was synthesized following the procedure described in preparative example C-90 (step 3) using the compound (101 mg, 0.23 mmol) obtained in (step 2) above.

[1487] MS m / z: 298 [M+l]+

[1488] [Preparative example C-1091 tert-butyl ( 1 -(3-amino-2-(azetidin- 1 -ylmethyl)propyl)- lH-pyrazol-4-yl)carbamate (Int. C-109)

[1489] Int. C-109

[1490] (Step 1) tert-butyl (l-(3-(((benzyloxy)carbonyl)amino)-2-formylpropyl)-lH-pyrazol-4- yl)carbamate

[1491] To a solution of the Int. C-107 (816 mg, 2.01 mmol) in DCM (10 mL) was added Dess- Martin periodinane (1.03 g, 2.42 mmol) carefully at 0 °C with stirring. After stirring at RT for 2 h under nitrogen atmosphere, the reaction was quenched by adding saturated NaHCOt aqueous solution (20 mL), then extracted three times with DCM (40 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (DCM : MeOH = 97 : 3) to obtain the title compound as a crystalline solid (801 mg, 99%).

[1492] MS m / z: 403 [M+l]+.1H NMR (400 MHz, CDCh) 5 9.78 (s, 1H), 7.64 (s, 1H), 7.44 - 7.30 (m, 6H), 6.36 (s, 1H), 5.48 (d, J = 6.5 Hz, 1H), 5.08 (s, 2H), 4.38 (d, J = 6.0 Hz, 2H), 3.52 - 3.31 (m, 2H), 3.05 (t, J = 5.8 Hz, 1H), 1.49 (s, 9H).

[1493] (Step 2) tert-butyl (l-(3-(azetidin-l-yl)-2-

[1494] ((((benzyloxy)carbonyl)amino)methyl)propyl)-lH-pyrazol-4-yl)carbamate

[1495] To a solution of the compound prepared in (step 2) above (250 mg, 0.62 mmol) in DCE (5 mL) were added azetidine (0.06 mL, 0.89 mmol) and acetic acid (0.07 mL, 1.22 mmol) at 0 °C. After stirring at RT for 20 min, NaBH(OAc)s (263 mg, 1.24 mmol) was added to the mixture at 0 °C. The reaction mixture was stirred at RT for 0.5 h under nitrogen atmosphere. On completion, the reaction was quenched by adding saturated NaHCO t aqueous solution (20

[1496] -148-

[1497] FH13103096.12 Attorney Docket No.: LCH-03225 mL), then extracted twice with DCM (40 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by MPLC (DCM : MeOH = 95 : 5 to 85 : 15) to obtain the title compound as a colorless solid (197 mg, 71%).

[1498] MS m / z: 444 [M+l]+

[1499] (Step 3) tert-butyl ( 1 -(3-amino-2-(azetidin- 1 -ylmethyl)propyl)- 1 H-pyrazol-4- yl)carbamate (Int. C-109)

[1500] The title compound (137 mg, 99%) was synthesized following the procedure described in preparative example C-90 (step 3) using the compound (197 mg, 0.44 mmol) obtained in (step 2) above.

[1501] MS m / z: 310 [M+l]+

[1502] [Preparative example C-1101 l-(3-amino-2-((dimethylamino)methyl)propyl)-lH- pyrazol-4-ol (Int. C-110)

[1503] (Step 1) 3-hydroxy-2-((4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazol-l- yl)methyl)propanenitrile

[1504] To a mixture of pyrazole-4-boronic acid pinacol ester (500 mg, 2.58 mmol) dissolved in ACN (5 mL) were added 2-(hydroxymethyl)prop-2-enenitrile (0.31 mL, 3.87 mmol) and l,8-diazabicyclo[5.4.0]-7-undecene (0.19 mL, 1.27 mmol). The reaction mixture was stirred at RT for 23 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as a colorless oil (339 mg, 47%).

[1505] MS m / z: 278 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.82 (s, 1H), 7.81 (s, 1H), 4.61 - 4.53 (m, 1H), 4.49 - 4.43 (m, 1H), 3.75 - 3.63 (m, 2H), 3.61 - 3.45 (m, 1H), 3.31 - 3.19 (m, 1H), 1.32 (s, 12H).

[1506] -149-

[1507] FH13103096.12 Attorney Docket No.: LCH-03225

[1508] (Step 2) tert-butyl (3-hydroxy-2-((4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrazol- 1 -yl)methyl)propyl)carbamate

[1509] To a solution of the compound prepared in (step 1) above (339 mg, 1.22 mmol) in a mixed solvent of MeOH / THF (12 mL, 3 : 1) were added Raney-Ni (339 mg) and 28-30% ammonia aqueous solution (0.6 mL). After stirring at RT for 22 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure. The residue was dissolved in DCM (12 mL) and were added BOC2O (0.28 mL, 1.22 mmol) and TEA (0.26 mL, 1.87 mmol). The reaction mixture was stirred at RT for 3 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 3 : 2) to obtain the tide compound as a white foam solid (196 mg, 42%).

[1510] MS m / z: 382 [M+l]+.1H NMR (400 MHz, CDCI3) 5 7.77 (s, 1H), 7.69 (s, 1H), 5.05 - 4.97 (m, 1H), 4.19 - 4.11 (m, 2H), 3.86 - 3.79 (m, 1H), 3.59 - 3.51 (m, 1H), 3.45 - 3.35 (m, 1H), 3.21 - 3.13 (m, 1H), 3.04 - 2.95 (m, 1H), 2.22 - 2.14 (m, 1H), 1.44 (s, 9H), 1.31 (s, 12H).

[1511] (Step 3) tert-butyl (3-hydroxy-2-((4-hydroxy-lH-pyrazol-l- yl)methyl)propyl)carbamate

[1512] To a solution of the compound prepared in (step 2) above (196 mg, 0.51 mmol) in THF (5 mL) were added 30% hydrogen peroxide solution (89 pL, 1.02 mmol) and NaOH (41.0 mg, 1.03 mmol) at 0 °C. The reaction mixture was stirred at RT for 1 hour. After completion of the reaction, the mixture was quenched by adding 1 N HC1 aqueous solution (20 mL), then extracted five times with 25% z-PrOH in chloroform (20 mL). The combined organic layers were dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 15) to obtain the title compound as a colorless oil (89.0 mg, 64%).

[1513] MS m / z: 272 [M+l]+.1H NMR (400 MHz, CD3OD) 5 7.21 (s, 1H), 7.08 (s, 1H), 6.70 - 6.61 (m, 1H), 4.06 - 3.98 (m, 2H), 3.45 - 3.39 (m, 2H), 3.10 - 3.00 (m, 2H), 2.15 - 2.07 (m, 1H), 1.44 (s, 9H).

[1514] (Step 4) l-(3-((tert-butoxycarbonyl)amino)-2-(((methylsulfonyl)oxy)methyl)propyl)- lH-pyrazol-4-yl methanesulfonate

[1515] To a solution of the compound prepared in (step 3) above (89.0 mg, 0.33 mmol) in DCM (4 mL) were added TEA (69 pL, 0.50 mmol) and methanesulfonyl chloride (25 pL, 0.33 mmol) at 0 °C. The mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (10 mL), then extracted with

[1516] -150-

[1517] FH13103096.12 Attorney Docket No.: LCH-03225

[1518] DCM (10 mL). The organic layer was dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The title compound was obtained as a light brown oil (167 mg), which was used for the next step without further purification.

[1519] MS m / z: 428 [M+l]+. ’ H NMR (400 MHz, CDCh) 5 7.61 (s, 1H), 7.51 (s, 1H), 5.04 - 4.94 (m, 1H), 4.19 - 4.08 (m, 4H), 3.24 - 3.15 (m, 2H), 3.13 (s, 3H), 3.06 (s, 3H), 2.60 - 2.54 (m, 1H), 1.44 (s, 9H).

[1520] (Step 5) 1 -(3-((tert-butoxycarbonyl)amino)-2-((dimethylamino)methyl)propyl)- 1H- pyrazol-4-yl methanesulfonate

[1521] The title compound (41.8 mg, 34%) was synthesized following the procedure described in preparative example C-108 (step 2) using the compound (140 mg, 0.33 mmol) obtained in (step 4).

[1522] MS m / z: 377 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.63 (s, 1H), 7.47 (s, 1H), 5.38 - 5.31 (m, 1H), 4.13 (d, J = 5.5 Hz, 2H), 3.19 - 3.14 (m, 1H), 3.11 (s, 3H), 2.89 - 2.80 (m, 1H), 2.29 - 2.23 (m, 1H), 2.20 (s, 6H), 2.15 - 2.08 (m, 1H), 2.05 - 1.97 (m, 1H), 1.44 (s, 9H).

[1523] (Step 6) tert-butyl (3-(dimethylamino)-2-((4-hydroxy-lH-pyrazol-l- yl)methyl)propyl)carbamate

[1524] To a solution of the compound prepared in (step 5) above (41.8 mg, 0.11 mmol) in a mixed solvent of THF / MeOH (2 mL, 1 : 1) was added 2 N NaOH aqueous solution (0.28 mL, 0.56 mmol). The reaction mixture was stirred at 60 °C for 4 h under nitrogen atmosphere. After being cooled to RT, the reaction was quenched by adding 4 N HC1 solution in 1,4-dioxane to adjust pH below 4, then concentrated under reduced pressure. The title compound was obtained as a yellow solid (86.0 mg), which was used for the next step without further purification.

[1525] MS m / z: 299 [M+l]+

[1526] (Step 7) l-(3-amino-2-((dimethylamino)methyl)propyl)-lH-pyrazol-4-ol (Int. C-110)

[1527] To a solution of the compound prepared in (step 6) above (33.1 mg, 0.111 mmol) in MeOH (1 mL) was added 4 N HC1 solution in 1,4-dioxane (1 mL). The reaction mixture was stirred at RT for 1 h under nitrogen atmosphere. On completion, the solvent was removed under reduced pressure. The title compound was obtained as a light brown solid (67.8 mg), which was used for the next step without further purification.

[1528] MS m / z: 199 [M+l]+

[1529] [Preparative example C-1111 3-(5 ,6-dihydro- 1 ,2,4-triazin- l(4H)-yl)propan- 1 -amine (Int. C-lll)

[1530] -151-

[1531] FH13103096.12 Attorney Docket No.: LCH-03225

[1532] (Step 1) tert-butyl 2-(3-(((benzyloxy)carbonyl)amino)propyl)hydrazine- 1 -carboxylate To a mixture of benzyl N-(3-bromopropyl)carbamate (500 mg, 1.84 mmol) dissolved in anhydrous DMF (6 mL) was added tert-butyl carbazate (728 mg, 5.51 mmol). The reaction mixture was stirred at 100 °C for 3 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (50 mL), then extracted with EA (50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as a colorless oil (383 mg, 64%).

[1533] MS m / z: 324 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.39 - 7.30 (m, 5H), 6.09 - 5.93 (m, 1H), 5.32 - 5.21 (m, 1H), 5.09 (s, 2H), 4.06 - 3.87 (m, 1H), 3.34 - 3.25 (m, 2H), 2.96 - 2.86 (m, 2H), 1.71 - 1.62 (m, 2H), 1.45 (s, 9H).

[1534] (Step 2) tert-butyl 2-(3-(((benzyloxy)carbonyl)amino)propyl)-2-(2-((tert- butoxycarbonyl)amino)ethyl)hydrazine- 1 -carboxylate

[1535] To a solution of the compound prepared in (step 1) above (383 mg, 1.18 mmol) in MeOH (5 mL) were added acetic acid (0.27 mL, 4.73 mmol) and N-Boc-2-aminoacetaldehyde (283 mg, 1.78 mmol) at RT. After stirring at the same temperature for 2 h, NaBH tCN (149 mg, 2.37 mmol) was added thereto, then the resulting mixture was stirred at RT for 15 h under nitrogen atmosphere. The reaction was quenched by adding saturated NaHCCL aqueous solution (30 mL), then extracted twice with EA (30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 2) to obtain the title compound as a colorless oil (308 mg, 56%).

[1536] MS m / z: 467 [M+l]+.1H NMR (400 MHz, CDCh) 57.40 - 7.28 (m, 5H), 5.82 (s, 1H), 5.55 - 5.43 (m, 1H), 5.30 - 5.25 (m, 1H), 5.10 (s, 2H), 3.37 - 3.29 (m, 2H), 3.24 - 3.15 (m, 2H), 2.82 - 2.63 (m, 4H), 1.73 - 1.64 (m, 2H), 1.45 - 1.42 (m, 18H).

[1537] (Step 3) benzyl (3-(l-(2-aminoethyl)hydrazineyl)propyl)carbamate

[1538] The title compound (412 mg) was synthesized following the procedure described in -152-

[1539] FH13103096.12 Attorney Docket No.: LCH-03225 preparative example C-92 (step 4) using the compound (308 mg, 0.66 mmol) obtained in (step 2) above.

[1540] MS m / z: 267 [M+l]+

[1541] (Step 4) benzyl (3 -(5 ,6-dihydro- 1 ,2,4-triazin- 1 (4H)-yl)propyl)carbamate

[1542] A mixture of the compound prepared in (step 3) above (120 mg, 0.24 mmol) and trimethyl orthoformate (80 pL, 0.73 mmol) dissolved in MeOH (0.5 mL) was stirred at 60 °C for 2 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with saturated NaHCOt aqueous solution (10 mL), then extracted with EA (10 mL). The organic layer was dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (MeOH : DCM = 1 : 9) to obtain the title compound as a colorless oil (20.5 mg, 31%).

[1543] MS m / z: 277 [M+l]+.1H NMR (400 MHz, CDCh) 57.37 - 7.28 (m, 5H), 6.76 (s, 1H), 5.49 - 5.36 (m, 1H), 5.08 (s, 2H), 4.26 - 4.13 (m, 1H), 3.49 - 3.41 (m, 2H), 3.37 - 3.28 (m, 2H), 2.89 - 2.82 (m, 2H), 2.79 - 2.72 (m, 2H), 1.90 - 1.84 (m, 2H).

[1544] (Step 5) 3-(5,6-dihydro-l,2,4-triazin-l(4H)-yl)propan-l-amine (Int. C-lll)

[1545] The title compound (lO.Omg) was synthesized following the procedure described in preparative example C-87 (step 3) using the compound (20.5 mg, 0.07 mmol) obtained in (step 4).

[1546] MS m / z: 143 [M+l]+[Preparative example C-1121 3-(5,6-dihydro-l,2,4-triazin-4(lH)-yl)propan-l-amine (Int. C- 112)

[1547] (Step 1) benzyl (3-((2-hydroxyethyl)amino)propyl)carbamate

[1548] To a mixture of benzyl N-(3-bromopropyl)carbamate (2.00 g, 7.35 mmol) dissolved in ACN (20 mL) were added ethanolamine (0.89 mL, 14.7 mmol) and K2CO3 (4.06 g, 29.4 mmol). The reaction mixture was stirred at 70 °C for 2 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (100 mL), then extracted with 10% MeOH -153-

[1549] FH13103096.12 Attorney Docket No.: LCH-03225 in DCM (100 mL). The organic layer was dried over anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The title compound was obtained as a colorless oil (1.69 g), which was used for the next step without further purification.

[1550] MS m / z: 253 [M+l]+

[1551] (Step 2) 2-(trimethylsilyl)ethyl (3-(((benzyloxy)carbonyl)amino)propyl)(2- hydroxyethyl)carbamate

[1552] To a solution of the compound prepared in (step 1) above (1.69 g, 6.70 mmol) in DCM (30 mL) were added N-[2-(trimethylsilyl)ethoxycarbonyloxy]succinimide (1.74 g, 6.70 mmol) and TEA (2.8 mL, 20.1 mmol). The reaction mixture was stirred at RT for 18 h under nitrogen atmosphere. On completion, the reaction mixture was diluted with distilled water (80 mL), then extracted twice with DCM (80 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, then concentrated under reduced pressure. The remaining residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as colorless oil (1.66 mg, 62%).

[1553] MS m / z: 397 [M+l]+.1H NMR (400 MHz, CDCh) 57.38 - 7.30 (m, 5H), 5.09 (s, 2H), 4.21 - 4.12 (m, 2H), 3.78 - 3.69 (m, 2H), 3.44 - 3.30 (m, 4H), 3.24 - 3.14 (m, 2H), 1.78 - 1.68 (m, 2H), 1.04 - 0.95 (m, 2H), 0.03 (s, 9H).

[1554] (Step 3) 2-(trimethylsilyl)ethyl (3-(((benzyloxy)carbonyl)amino)propyl)(2- oxoethyl)carbamate

[1555] The title compound (516 mg, 62 %) was synthesized following the procedure described in preparative example C-42 (step 2) using the compound (836 mg, 2.11 mmol) obtained in (step 2) above.

[1556] MS m / z: 395 [M+l]+.1H NMR (400 MHz, CDCh) 59.58 (s, 1H), 7.37 - 7.31 (m, 5H), 5.09 (s, 2H), 4.21 - 4.12 (m, 2H), 4.04 - 3.94 (m, 2H), 3.42 - 3.33 (m, 2H), 3.27 - 3.20 (m, 2H), 1.74 - 1.61 (m, 2H), 1.06 - 0.91 (m, 2H), 0.02 (s, 9H).

[1557] (Step 4) tert-butyl 1 l-oxo-13-phenyl-6-((2-(trimethylsilyl)ethoxy)carbonyl)-12-oxa- 2 , 3 ,6 , 10-tetraazatridecanoate

[1558] To a solution of the compound prepared in (step 3) above (516 mg, 1.31 mmol) in MeOH (12 mL) were added tert-butyl carbazate (173 mg, 1.31 mmol), acetic acid (90 pL, 1.57 mmol) and NaB H tCN (123 mg, 1.96 mmol) at 0 °C. After stirring at RT for 21 h under nitrogen atmosphere, the reaction was quenched by adding saturated NaHCCh aqueous solution (50 mL), then extracted twice with DCM (50 mL). The combined organic layers were dried over

[1559] -154-

[1560] FH13103096.12 Attorney Docket No.: LCH-03225 anhydrous Na2SC>4, filtered, then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (EA : n-hexane = 1 : 1) to obtain the title compound as a colorless oil (355 mg, 53%).

[1561] MS m / z: 511 [M+l]+. ’ H NMR (400 MHz, CDCh) 57.38 - 7.30 (m, 5H), 5.09 (s, 2H), 4.19 - 4.13 (m, 2H), 3.38 - 3.27 (m, 4H), 3.22 - 3.15 (m, 2H), 2.99 - 2.93 (m, 2H), 1.76 - 1.67 (m, 2H), 1.45 (s, 9H), 1.03 - 0.98 (m, 2H), 0.03 (s, 9H).

[1562] (Step 5) benzyl (3-((2-hydrazineylethyl)amino)propyl)carbamate

[1563] To a solution of the compound prepared in (step 4) above (355 mg, 0.70 mmol) in DCM (8 mL) was added TFA (4 mL). The mixture was stirred at RT for 2 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The title compound was obtained as a light yellow oil (381 mg), which was used for the next step without further purification.

[1564] MS m / z: 367 [M+l]+

[1565] (Step 6) benzyl (3-(l-formyl-5,6-dihydro-l,2,4-triazin-4(lH)-yl)propyl)carbamate

[1566] To a solution of the compound prepared in (step 5) above (50.0 mg, 0.15 mmol) in acetic acid (1 mL) was added trimethyl orthoformate (0.50 mL). The mixture was stirred at 100 °C for 3 h under nitrogen atmosphere. On completion, the reaction mixture was concentrated under reduced pressure. The title compound was obtained as a yellow oil (43.5 mg), which was used for the next step without further purification.

[1567] MS m / z: 305 [M+l]+.1H NMR (400 MHz, CDCh) 5 8.45 (s, 1H), 7.38 - 7.34 (m, 5H), 6.63 (s, 1H), 5.10 (s, 2H), 4.85 - 4.76 (m, 1H), 3.85 - 3.77 (m, 2H), 3.30 - 3.15 (m, 6H), 1.82 - 1.72 (m, 2H).

[1568] (Step 7) 3-(5,6-dihydro-l,2,4-triazin-4(lH)-yl)propan-l-amine (Int. C-112)

[1569] To a solution of the compound prepared in (step 6) above (43.5 mg, 0.14 mmol) in MeOH / EA (2 mL, 1 : 1) were added 4 N HC1 solution in 1,4-dioxane (0.2 mL) and Pd / C (13.0 mg). After stirring at RT for 23 h under hydrogen atmosphere (1 atm), the reaction mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to obtain the title compound as a light brown oil (42.5 mg) which was used for the next step without further purification.

[1570] MS m / z: 143 [M+l]+

[1571] Preparation Example 4: Synthesis of intermediate D series

[1572] [Preparative example D-l] 2-(3-(4-amino- IH-pyrazol- 1 -yl)propyl)isoindoline- 1 ,3-dione (Int.

[1573] -155-

[1574] FH13103096.12 Attorney Docket No.: LCH-03225

[1575] D-l)

[1576] The title compound (274 mg, 94%) was synthesized following the procedure described in preparative example C- 105 (step 1) using tert-butyl (l-(3-(l ,3-dioxoisoindolin-2-yl)propyl)- lH-pyrazol-4-yl)carbamate (400 mg, 1.08 mmol).

[1577] MS m / z: 271 [M+l]+.1H NMR (400 MHz, CDCh) 5 7.86 - 7.79 (m, 2H), 7.74 - 7.67 (m, 2H), 7.10 (s, 2H), 4.09 - 4.00 (m, 2H), 3.75 - 3.66 (m, 2H), 2...

Claims

Attorney Docket No.: LCH-03225We claim:

1. A compound having a structure represented by Formula I, or a pharmaceutically acceptable salt thereof:whereinR1, R2, R3and R4are each independently H, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, sulfonyl, cycloalkyl, or heterocyclyl; or R2and R3, together with the carbon atoms that separate them, complete a heterocyclyl;Y is alkylene, alkenylene, alkynylene, cycloalkylene, heteroalkylene, or heteroalkenyleneZ is aryl, heteroaryl, cycloalkyl, heterocyclyl, amido, or amino; andA is aryl or heteroaryl.

2. The compound of claim 1, having a structure represented by Formula la, or a pharmaceutically acceptable salt thereof:whereinR1, R2, R3and R4are each independently H, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, cycloalkyl, or heterocyclyl; or R2and R3, together with the carbon atoms that separate them, complete a heterocyclyl;Y is alkylene, alkenylene, cycloalkylene, heteroalkylene, or heteroalkenyleneZ is aryl, heteroaryl, cycloalkyl, heterocyclyl, amido, or amino; and-304-FH13103096.12Attorney Docket No.: LCH-03225A is aryl or heteroaryl.

3. The compound of claim 1 or 2, wherein if R2and R3are both H or both methoxy and Z is heteroaryl or heterocyclyl, then Z is substituted with -NRaRbor -alkyl-NRaRb, wherein Raand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

4. The compound of claim 1 or 2, wherein if R2is nitro or halo and Z is heteroaryl or heterocyclyl, then Z is substituted with -NRaRbor - alkyl-NRaRb.

5. The compound of claim 1 or 2, wherein if R2is methoxy and R3is H, then Z is aryl, heteroaryl, cycloalkyl, amido, or amino.

6. The compound of claim 1 or 2, wherein if R2or R3is carboxyl and Z is heteroaryl or heterocyclyl, then Z is substituted with -NRaRbor -alkyl-NRaRb.

7. The compound of claim 1 or 2, wherein A is aryl (preferably phenyl).

8. The compound of claim 1 or 2, wherein A is heteroaryl (e.g., pyridinyl).

9. The compound of any one of claims 1-8, wherein the compound has a structure represented by Formula lb, or a pharmaceutically acceptable salt thereof:whereinX is CH or N; andFH13103096.12Attorney Docket No.: LCH-03225R5and R6are each independently H, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, cycloalkyl, or heterocyclyl; or R5and R6, together with the carbon atoms that separate them, complete a heterocyclyl.

10. The compound of claim 9, wherein X is CH.

11. The compound of claim 9, wherein X is N.

12. The compound of any one of claims 9-11, wherein R5is alkyl (e.g., methyl).

13. The compound of any one of claims 9-11, wherein R5is halo (e.g., fluoro).

14. The compound of any one of claims 9-11, wherein R5is alkoxy (preferably methoxy).

15. The compound of claim 14, wherein R6is halo (preferably fluoro).

16. The compound of any one of claims 9-14, wherein R6is alkoxy (preferably methoxy).

17. The compound of any one of claims 9-14, wherein R6is alkyl (e.g., methyl).

18. The compound of any one of claims 9-14, wherein R6is H.

19. The compound of claim 16, wherein R5is halo (preferably fluoro).

20. The compound of claim 9, wherein R5and R6, together with the carbon atoms that separate them, complete a heterocycle (e.g., 1,3 -dioxolane).

21. The compound of any one of claims 1-20, wherein the compound has a structure represented by Formula Ic or a pharmaceutically acceptable salt thereof:-306-FH13103096.12Attorney Docket No.: LCH-03225Ic whereinR7and R8are each independently H, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, cycloalkyl, or heterocyclyl.

22. The compound of claim 21, wherein the compound has a structure represented by Formula Id or a pharmaceutically acceptable salt thereof:Id whereinR3is methoxy or cyano; andR2is halo (preferably fluoro).

23. The compound of claim 21 or 22, wherein R7and R8are each deuterium.

24. The compound of any one of claims 21-23, wherein R7and R8are each independently H, Ci-6 alkyl, Ci-6 alkoxy, C2-6 alkenyl, C2-6 alkynyl, or fluoro.

25. The compound of any one of claims 1-24, wherein Z is heteroaryl (e.g., thiazolyl, pyrazolyl, pyridazinyl, imidazolyl, pyridyl, pyrimidyl, pyrazinyl, or triazinyl, each of which is optionally substituted).

26. The compound of any one of claims 1-24, wherein Z is heterocyclyl (e.g., pyrrolidinyl, piperidinyl, piperidinonyl, piperazyl, imidazolidinyl, oxazolidinyl, isothiazolidinyl, hexahydropyridazinyl, morpholinyl, tetrahydropyrrolopyridyl, tetrahydropyrrolopyridinonyl, pyranopyrimidinedionyl, pyrrolopyrimidyl, oxazaspirodecyl, or oxadiazolyl, each of which is optionally substituted).-307-FH13103096.12Attorney Docket No.: LCH-0322527. The compound of any one of claims 1-26, wherein Z is substituted with oxo, amido, alkyl, halo, or hydroxyl, and is optionally further substituted.

28. The compound of any one of claims 1-25, wherein Z is pyrazolyl.

29. The compound of any one of claims 1-23, wherein Z is dihydropyranopyridinedionyl30. The compound of any one of claims 1-29, wherein:Z is heteroaryl or heterocyclyl;Z is substituted with -NRaRbor -alkyl-NRaRb, and is optionally further substituted; andRaand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

31. The compound of any one of claims 1-24, wherein Z is cycloalkyl (e.g., cyclobutyl, cyclopentyl, bicyclopentyl, or bicyclohexyl, each of which is optionally substituted).

32. The compound of any one of claims 1-24, wherein Z is cycloalkyl (e.g., bicyclo[l .

1. l]pentanyl or bicyclo[2.

1. l]hexanyl).

33. The compound of any one of claims 1-24, wherein Z is amido.

34. The compound of any one of claims 1-24, wherein Z is substituted or unsubstituted alkanecarboxamido (e.g., morpholinylacetamido, hydroxyacetamido, or aminoacetamido).-308-FH13103096.12Attorney Docket No.: LCH-0322535. The compound of any one of claims 1-24, wherein Z is substituted or unsubstituted alkanecarboxamido (e.g., morpholinylacetamido or hydroxyacetamido).

36. The compound of any one of claims 1-24, wherein Z is alkyl-substituted carboxamido.

37. The compound of any one of claims 1-36, wherein Z is substituted with alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, hydroxyl, carboxyl, acyl, ester, halo, thioester, amino, amido, cyano, nitro, cycloalkyl, and heterocyclyl.

38. The compound of any one of claims 1-36, wherein Z is substituted with hydroxyl, halo (e.g., fluoro), carboxyl, alkoxy (e.g., methoxy), amido (e.g., hydroxymethylamido), sulfonamidyl (e.g., methyl sulfonamidyl), amino, or alkylamino (e.g., methylamino, ethylamino).

39. The compound of any one of claims 1-36, wherein Z is substituted with alkyl (e.g., (fluoro)(hydroxy)methyl or aminoalkyl).

40. The compound of any one of claims 1-39, wherein Y is alkylene (e.g., propylene or butylene), optionally substituted with hydroxyl or dimethylaminopropyl.

41. The compound of claim 40, wherein Y is alkylene (e.g., propylene or butylene) substituted with hydroxyl, dimethylaminomethyl, dimethylaminopropyl, azetidinemethyl,42. The compound of claim 40 or 41, wherein Y is -(CH2)t-, wherein t is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

43. The compound of claim 42, wherein t is 3.

44. The compound of claim 42, wherein t is 4.

45. The compound of any one of claims 1-39, wherein Y is heteroalkylene.-309-FH13103096.12Attorney Docket No.: LCH-0322546. The compound of claim 45, wherein Y is *-NHCH2CH2-, *-CH2CH2-NH-, *- CH2CH2O-, *-CH2CH2CH2O-, or *-CH2CH2CH2NH-, wherein * indicates the point of attachment to the nitrogen of formula I.

47. The compound of any one of claims 1-39, wherein Y is alkenylene (e.g., butenyl-2- ene).

48. The compound of any one of claims 1-39, wherein Y is cycloalkylene (e.g., methylcyclobutylene) .

49. The compound of any one of claims 1-48, wherein Y is substituted with alkyl (e.g. , methyl).

50. The compound of any one of claims 1-49, wherein Y is substituted with alkyl (e.g. , N,N-dimethylaminoethyl or N,N-dimethylaminomethyl).

51. The compound of any one of claims 1-50, wherein Y is substituted with amido (e.g., aminoalkylaminoalkylamido or aminoalkoxyalkylamido).

52. The compound of claim 49, wherein Y is substituted with two methyls.

53. The compound of any one of claims 1-49, wherein Y is further substituted with two fluoros.

54. The compound of any one of claims 1-53, wherein Y is substituted with hydroxyl.

55. The compound of any one of claims 1-53, wherein Y is disubstituted, preferably on the same carbon, and the substituents combine to form a heterocyclyl (e.g., an oxetanyl).

56. The compound of any one of claims 1-40, wherein the compound has a structure represented by Formula le or a pharmaceutically acceptable salt thereof:-310-FH13103096.12Attorney Docket No.: LCH-03225whereinT is alkyl (e.g., hydroxyalkyl, (hydroxy)(halo)alkyl, cycloalkylalkyl, or heterocyclylalkyl, such as morpholinylalkyl), andR9is H, alkyl (e.g., trifluoroethyl, dimethylaminoethyl), aralkyl (e.g., benzyl or phenylethyl), or cycloalkyl (e.g., cyclopropyl).

57. The compound of any one of claims 1-40, wherein the compound has a structure represented by Formula le or a pharmaceutically acceptable salt thereof:whereinT is alkyl (e.g., hydroxyalkyl, (hydroxy)(halo)alkyl, cycloalkylalkyl, or heterocyclylalkyl, such as morpholinylalkyl), andR9is H, alkyl (e.g., trifluoroethyl), aralkyl (e.g., benzyl or phenylethyl), or cycloalkyl (e.g., cyclopropyl).

58. The compound of claim 56 or 57, wherein T is alkyl (e.g., methyl) substituted with alkoxy (e.g., methylaminoethoxy), alkylamino (e.g., aminoethylmethylamino, dimethylaminoethylamino), or aminocycloalkyl (e.g., aminocyclopropyl).

59. The compound of claim 56 or 57, wherein T is alkyl (e.g., trifluoroethyl, difluoroethyl, propyl, ethyl, methyl) substituted with one, two, or three substituents selected from amino, (e.g., dimethylamino), hydroxyl, aryl (e.g., phenyl), and halo (e.g., fluoro).-311-FH13103096.12Attorney Docket No.: LCH-0322560. The compound of any one of claims 1-55, wherein the compound has a structure represented by Formula If or a pharmaceutically acceptable salt thereof:wherein n ranges from 0 to 5; p ranges from 0 to 3;Z is aryl, heteroaryl, cycloalkyl, heterocyclyl, amido, or amino; andRaand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, heteroaryl, alkylsulfoxidyl, alkylsulfonyl, or sulfonamido.

61. The compound of claim 60, wherein n is 0.

62. The compound of claim 60, wherein n is 1.

63. The compound of any one of claims 1-40, wherein the compound has a structure represented by Formula Ig or a pharmaceutically acceptable salt thereof:whereinXi is C, CH, or N;X2, X3, X4 and X5 are each independently CH, CH2, N, NH, O, or S; at least two of Xi, X2, X3, X4 or X5 are C, CH or CH2;-312-FH13103096.12Attorney Docket No.: LCH-03225 each of Xi, X2, X3, X4 and X5 is optionally substituted with W; each W is independently selected from alkyl, hydroxyl, oxo, amido, amino, heteroaryl, and heterocyclyl; is a single bond or a double bond; and m is an integer selected from 1 and 2.

64. The compound of claim 63, wherein: at least one W is NRaRb; andRaand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, amido, or alkylsulfonyl.

65. The compound of claim 63 or 64, wherein m is 1.

66. The compound of any one of claims 63-65, wherein Xi, X3 and X5 are N; and X2 and X4 are CH, optionally substituted with W.

67. The compound of any one of claims 63-66, wherein the compound has a structure, represented by Formula Ih or a pharmaceutically acceptable salt thereof:whereinU is alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

68. The compound of any one of claims 63-66, wherein the compound has a structure, represented by Formula li or a pharmaceutically acceptable salt thereof:-313-FH13103096.12Attorney Docket No.: LCH-03225whereinU is NRaRb, wherein Raand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

69. The compound of any one of claims 63-68, wherein the compound has a structure, represented by Formula Ij or a pharmaceutically acceptable salt thereof:

70. The compound of any one of claims 63-65, wherein two of Xi, X2, X3, X4 and X5 areN or NH, and the remainder are C or CH, optionally substituted with W.

71. The compound of any one of claims 63-65, wherein Xi and X2 are N; and X3, X4, and X5 are CH, optionally substituted with W.

72. The compound of any one of claims 63-65, 70, and 71, wherein the compound has a structure, represented by Formula Ik or a pharmaceutically acceptable salt thereof:-314-FH13103096.12Attorney Docket No.: LCH-03225Ik whereinU is alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

73. The compound of any one of claims 63-65, 70, and 71, wherein the compound has a structure, represented by Formula II or a pharmaceutically acceptable salt thereof:II whereinU is hydroxyl or NRaRb, wherein Raand Rbare each independently H, alkyl, alkoxy, alkenyl, alkynyl, aralkyl, heteroaralkyl, aryl, heteroaryl, carboxyl, acyl, amido, ester, thioester, cycloalkyl, heterocyclyl, alkylsulfoxidyl, or alkylsulfonyl.

74. The compound of any one of claims 64-73, wherein U is amido (e.g., aminoalkylamido, aminoalkylaminoalkylamido, carboxyalkylamido, amidoalkylamido, cycloalkylamido, such as aminocycloalkylamido, or hydrazineamido).

75. The compound of any one of claims 63-65 and 70-73, wherein the compound has a structure, represented by Formula Im or a pharmaceutically acceptable salt thereof:

76. The compound of claim 69 or 75, wherein Rais H.

77. The compound of claim 69 or 75, wherein Rais alkyl (e.g., methyl or trifluoroethyl).-315-FH13103096.12Attorney Docket No.: LCH-0322578. The compound of claim 69 or 75, wherein Rais alkyl (e.g., ethyl).

79. The compound of any one of claims 74-77, wherein Rbis heterocyclyl (e.g., piperidinyl or pyrrolidinyl).

80. The compound of any one of claims 69 and 75-78, wherein Rbis alkyl (e.g., methyl, ethyl, propyl, butyl, or pentyl).

81. The compound of any one of claims 69 and 75-78, wherein Rbis alkyl (e.g., cyclobutylmethyl, cyclopentylmethyl or cyclobutylethyl).

82. The compound of any one of claims 75-78, wherein Rbis alkylsulfonyl (e.g., methylsulfonyl).

83. The compound of any one of claims 75-78, wherein Rbis heteroaryl (e.g. , pyridinyl).

84. The compound of any one of claims 75-78, wherein Rbis amido further substituted with amino, such as alkylamino (e.g., Rbis alkylN(H)N(H)C(O)- or methylN(H)N(H)C(O)-).

85. The compound of any one of claims 69 and 75-77, wherein Rbis acyl.

86. The compound of any one of claims 69 and 75-77, wherein Rbis polyethylene glycolyl (PEG).

87. The compound of any one of claims 75-77, wherein Rbis H.

88. The compound of any one of claims 69 and 75-87, wherein Rbis optionally substituted with one or more substituents selected from alkyl, hydroxyl, oxo, halo, alkoxy, and amino.

89. The compound of any one of claims 69 and 75-88, wherein Rbis optionally substituted with one, two, or three substituents selected from alkyl (e.g., hydroxymethyl), hydroxyl, oxo, halo, alkoxy, amino (e.g., dimethylaminoethylamino), alkylamino (e.g., dimethylamino, methylamino), cycloalkyl (e.g., cyclopropyl), amido (e.g., methylamido), and heterocycloalkyl (e.g., azetidinyl or oxetanyl).-316-FH13103096.12Attorney Docket No.: LCH-0322590. The compound of any one of claims 69 and 75-89, wherein Rbis optionally substituted with one, two, or three selected from alkyl, hydroxyl, oxo, halo, alkoxy, and amino.

91. The compound of claim 90, wherein the heterocycloalkyl on Rbis substituted with halo, hydroxyl, alkyl (e.g., trifluoromethyl), carboxyl, amino, or amido (e.g., methylamido).

92. The compound of any one of claims 88-91, wherein Rbis substituted with alkyl (e.g., methyl, or trifluoromethyl).

93. The compound of any one of claims 88-91, wherein Rbis substituted with alkyl (e.g., cyclopropylmethyl) and amino.

94. The compound of any one of claims 88-91, wherein Rbis substituted with hydroxyl.

95. The compound of any one of claims 88-91, wherein Rbis substituted with oxo.

96. The compound of any one of claims 88-91 , wherein Rbis substituted with halo (e.g. , fluoro).

97. The compound of any one of claims 88-91, wherein Rbis substituted with alkoxy (e.g., methoxy).

98. The compound of any one of claims 88-91, wherein Rbis substituted with aminoalkoxy.

99. The compound of any one of claims 88-91, wherein Rbis substituted with amino.

100. The compound of any one of claims 88-91, wherein Rbis substituted with hydroxyl and halo (e.g., fluoro).

101. The compound of any one of claims 88-91, wherein Rbis substituted with hydroxyl and alkyl (e.g., methyl, trifluoromethyl, cyclobutylmethyl, or cyclopropylmethyl).-317-FH13103096.12Attorney Docket No.: LCH-03225102. The compound of any one of claims 88-91, wherein Rbis substituted with oxo and alkoxy (e.g., methoxy).

103. The compound of any one of claims 88-91, wherein Rbis substituted with oxo and hydroxyl.

104. The compound of any one of claims 88-91, wherein Rbis substituted with hydroxyl, cycloalkyl (e.g., cyclopentyl) and halo (e.g., fluoro).

105. The compound of any one of claims 88-91, wherein Rbis substituted with hydroxyl and cycloalkyl (e.g., cyclopropyl).

106. The compound of any one of claims 88-91, wherein Rbis substituted with amino and halo (e.g., difluoro).

107. The compound of any one of claims 88-91, wherein Rbis substituted with amino, hydroxyl, and alkyl (e.g., methyl).

108. The compound of any one of claims 88-91, wherein Rbis substituted with amino and cycloalkyl (e.g., cyclobutyl, cyclopentyl, or cyclopropyl).

109. The compound of any one of claims 88-91, wherein Rbis substituted with alkyl (e.g., trifluoromethyl or dimethylaminopropyl) and amino.

110. The compound of any one of claims 1-109, wherein R2and R3are each independently alkoxy (e.g., methoxy, difluoromethoxy, ethoxy, difluoroethoxy, trifluoroethoxy, isopropoxy, or carboxymethoxy), halo (e.g., fluoro or chloro), or cyano.

111. The compound of claim 110, wherein R2is halo (preferably fluoro).

112. The compound of claim 110 or 111 , wherein R3is alkoxy (preferably methoxy, difluoromethoxy, or difluoroethoxy).

113. The compound of any one of claims 110-112, wherein R2is halo (preferably fluoro) and R3is methoxy.-318-FH13103096.12Attorney Docket No.: LCH-03225114. The compound of claims 110 or 111 , wherein R2is halo (preferably fluoro) and R3is cyano.

115. The compound of any one of claims 1-109, wherein R2and R3are each independently H or alkoxy (e.g., methoxy).

116. The compound of any one of claims 1-109, wherein R2and R3are each independently H or halo (e.g., fluoro).

117. The compound of any one of claims 1-109, wherein R2and R3are each independently alkoxy (e.g., methoxy or difluoromethoxy).

118. The compound of any one of claims 1-109, wherein R2and R3are each independently halo (e.g., fluoro or chloro) or cyano.

119. The compound of any one of claims 1-109, wherein R2and R3are each independently alkoxy (e.g., methoxy or difluoromethoxy) or cyano.

120. The compound of any one of claims 1-109, wherein R2and R3are each independently hydroxyl or cyano.

121. The compound of any one of claims 1-109, wherein R2and R3are each independently alkoxy (e.g., methoxy) or amino (e.g., -NH2 or difluoroethylamino).

122. The compound of any one of claims 1-109, wherein R2and R3are each independently alkoxy (e.g., methoxy) or nitro.

123. The compound of any one of claims 1-109, wherein R2and R3are each independently alkoxy (e.g., methoxy) or amido (e.g., acetamido or difluoroacetamido).

124. The compound of any one of claims 1-109, wherein R2and R3are each independently alkoxy (e.g., methoxy) or alkyl (e.g., trifluoromethyl).

125. The compound of any one of claims 1-109, wherein R2and R3are each independently halo (e.g., fluoro) or alkyl (e.g., trifluoromethyl).-319-FH13103096.12Attorney Docket No.: LCH-03225126. The compound of any one of claims 1-109, wherein R2and R3are each independently halo (e.g., fluoro) or hydroxyl.

127. The compound of any one of claims 1-109, wherein R2and R3are each independently halo (e.g., fluoro) or amido.

128. The compound of any one of claims 1-109, wherein R2and R3combine to form a heterocycle (e.g., 1,3-dioxolane).

129. The compound of any one of the preceding claims, wherein the compound is selected from:-320-FH13103096.12Attorney Docket No.: LCH-03225-321-FH13103096.12Attorney Docket No.: LCH-03225-322-FH13103096.12Attorney Docket No.: LCH-03225-323-FH13103096.12Attorney Docket No.: LCH-03225-324-FH13103096.12Attorney Docket No.: LCH-03225FH13103096.12Attorney Docket No.: LCH-03225FH13103096.12Attorney Docket No.: LCH-03225-327-FH13103096.12Attorney Docket No.: LCH-03225-328-FH13103096.12Attorney Docket No.: LCH-03225-329-FH13103096.12Attorney Docket No.: LCH-03225-330-FH13103096.12Attorney Docket No.: LCH-03225130. The compound of any one of claims 1-128, wherein the compound is selected from:FH13103096.12Attorney Docket No.: LCH-03225-332-FH13103096.12Attorney Docket No.: LCH-03225FH13103096.12Attorney Docket No.: LCH-03225-334-FH13103096.12Attorney Docket No.: LCH-03225FH13103096.12Attorney Docket No.: LCH-03225FH13103096.12Attorney Docket No.: LCH-03225-337-FH13103096.12Attorney Docket No.: LCH-03225FH13103096.12Attorney Docket No.: LCH-03225131. A pharmaceutical composition comprising the compound of any one of claims 1-130 and a pharmaceutically acceptable excipient.

132. A method of inhibiting topoisomerase I in a subject, comprising administering a compound of any one of claims 1-130, or a pharmaceutically acceptable salt thereof, to the subject.

133. A method of treating a disease modulated by topoisomerase I in a subject, comprising administering a compound of any one of claims 1-130, or a pharmaceutically acceptable salt thereof, to the subject.

134. A method of treating a proliferative disorder in a subject, comprising administering a compound of any one of claims 1-130, or a pharmaceutically acceptable salt thereof, to the subject.

135. The method of claim 134, wherein the proliferative disorder is cancer.

136. The method of claim 134 or 135, wherein the proliferative disease is selected from breast cancer, lung cancer, bladder cancer, brain cancer, head and neck cancer, liver cancer, biliary tract cancer, cervical cancer, endometrial cancer, esophageal cancer, carcinomas, acute and chronic lymphoid leukemias, acute and chronic myelogenous leukemias, chronic myelomonocytic leukemias, colorectal cancer, gastric cancer, gastrointestinal stromal cancer, glioma, lymphomas, melanomas, multiple myeloma, myeloproliferative diseases, neuroendocrine diseases, pancreatic cancer, ovarian cancer, prostate cancer, renal cell cancer, sarcomas, thyroid cancer, mast cell leukemia, germ cell tumors, testicular cancer, small-cell lung carcinoma, neuroblastoma, osteosarcoma, and retinoblastoma.-339-FH13103096.12Attorney Docket No.: LCH-03225137. The method of claim 136, wherein the proliferative disease is selected from breast cancer, lung cancer, bladder cancer, brain cancer, head and neck cancer, liver cancer, biliary tract cancer, carcinomas, acute and chronic lymphoid leukemias, acute and chronic myelogenous leukemias, chronic myelomonocytic leukemias, colorectal cancer, gastric cancer, gastrointestinal stromal cancer, glioma, lymphomas, melanomas, multiple myeloma, myeloproliferative diseases, neuroendocrine diseases, pancreatic cancer, ovarian cancer, prostate cancer, renal cell cancer, sarcomas, thyroid cancer, mast cell leukemia, germ cell tumors, small-cell lung carcinoma, neuroblastoma, and osteosarcoma.

138. A method of inhibiting topoisomerase I in a cell in vitro, comprising contacting the cell with a compound of any one of claims 1-130 or a pharmaceutically acceptable salt thereof.-340-FH13103096.12

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