Nampt inhibitor and antibody–drug conjugate containing same

Abstract

The present invention relates to an immunoconjugate comprising a cytotoxic drug having NAMPT inhibitory activity and, specifically, to a drug-linker conjugate comprising a cytotoxic drug having NAMPT inhibitory activity and a linker, an immunoconjugate comprising the drug-linker conjugate and an antibody or antigen-binding fragment thereof, and a composition for preventing or treating cancer, comprising the immunoconjugate or a pharmaceutically acceptable salt thereof. Exhibiting an excellent anticancer effect compared to a cytotoxic drug or antibody alone, with almost no side effects, the immunoconjugate of the present invention can be very advantageously used for preventing or treating cancer.

Description

[0001] Description of the Invention

[0002]

Title of invention

[0003] NAMPT inhibitors and antibody-drug conjugates containing the same

[0004]

Technical Field

[0005]

Background Technology

[0006] Nicotinamide phosphoribosyl transferase (NAMPT) catalyzes the synthesis of nicotinamide mononucleotide (NMN) from nicotinamide (NAM) and 5' phosphoribosyl-1'-pyrophosphate (PRPP), playing a key role in the cyclic biosynthetic pathway of NAD+ (nicotinamide adenine dinucleotide). NAD is essential for several signaling pathways, including mono-ADP-ribosylation in both the immune system and G-protein-coupled receptor signaling, among other poly-ADP-ribosylation in DNA repair, and is also essential for the deacetylase activity of sirtuins. NAD+ is produced by two distinct biosynthetic pathways, the salvage and de novo synthesis pathways. As a rate-limiting enzyme in the NAD+ salvage pathway, NAMPT is biologically essential and may be implicated in a number of diverse diseases. In particular, cancer cells must continuously resynthesize NAD+ to avoid NAD+ depletion and subsequent cell death. Increased expression of NAMPT has been reported in various cancers, including colon, breast, and ovarian cancers (Garten, A. et al., Nat. Rev. Endocrinol. 2015, 11:535—546; Sampath, D. et al., Pharmacol. Ther. 2015, 151:16-31). However, various NAMPT inhibitors have shown disappointing results in clinical trials due to dose-limiting toxicities, such as thrombocytopenia, retinal, and cardiac toxicity (Ghanem, MS et al., Nutrients, 2021, 13:1665).

[0007]

Contents of the invention

[0008]

Technical Challenges

[0009] Antibody-drug conjugates (ADCs) are immunoconjugates that combine a drug and an antibody via a linker. Because the antibody transports the drug to the target site and releases the drug, they are attracting attention as a new class of drugs due to their targeted nature and fewer side effects compared to single-agent use. The present inventors developed a payload with novel NAMPT inhibitory activity and, by applying it to an ADC, confirmed its excellent anticancer effect, thereby completing the present invention.

[0010]

Technical Solution

[0011] (1) The immunoconjugate or a pharmaceutically acceptable salt thereof according to the present invention is represented by the following Chemical Formula I:

[0012] [Chemical Formula I]

[0013] Ab-(L-D) p In Chemical Formula I above, 日 is a cytotoxic drug moiety represented by the following Chemical Formula III,

[0014] [Chemical Formula III] In Chemical Formula III above,

[0015] Ri is pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, pyrrolyl, H

[0016] ..、..、쒀 쇼〜、 4 i s

[0017] V- •쳟쇼

[0018] / ', and at this time, the above R1 의 내는 치환되지 않거나 적어도 하나의 - H7} 각각 독립적으로 - NH2, - (C1-3알킬) NH2, - NH(CI-3알킬), - N(CI-3알킬) 2, - 0H, - NO2, Ci-5알킬, - F, -Cl, -Br, 또는 - 1로치환되며, (The content in this part seems to be incomplete or contains some unclear notations and cannot be accurately translated. It should be noted that in the original text, there are some characters that may be encoding or input errors. For example, "의 내는 치환되지 않거나 적어도 하나의 - H7}" is difficult to understand clearly. But the translation is done as much as possible based on the existing text.) is unsubstituted or at least one of -H7 is independently substituted with -NH2, -(C1-3alkyl)NH2, -NH(CI-3alkyl), -N(CI-3alkyl)2, -OH, -NO2, Ci-5alkyl, -F, -Cl, -Br, or -1,

[0019] R2 is pyridinyl, pyrimidinyl, phenyl, piperidinyl, piperazinyl, morpholinyl, pyrazolyl, imidazolyl, phenolic, -CH2C(=0)NH2, -C(=0)NH2, -C(=0)NHCH2CH3, -CH2NHC(=0)CH2CH3, -NH2, -NHCH3, -NHCH2CH2CH3, It should be noted that there are some inaccuracies or unclear parts in the original text, which may affect the accuracy of the translation. It is recommended to check and correct the original text for a more accurate translation result.

[0020] At this time, the inside of the above R2 is independently unsubstituted or at least one of the insides is independently - NH2, - NH (CI-5 alkyl), - MC1-5 alkyl) 2, -NHCHs, -N (CH3) 2, - NHCH2CH3, -OH, - NO2, - S (= 0) 2CH3, - C1-5 alkyl (the inside of the above alkyl is unsubstituted or at least one of the insides is independently substituted with one selected from the group consisting of phenyl, pyridinyl, morpholinyl, piperazinyl and piperidinyl), - (Ci-5 alkyl) NH2, -morpholinyl, -piperidinyl, -piperazinyl, -phenyl, -pyridinyl, -pyrimidinyl (the insides of the above -morpholinyl, - piperidinyl, -piperazinyl, -phenyl, -pyridinyl and -pyrimidinyl are unsubstituted or at least one of the insides is independently - C1-3 alkyl and - CF3 substituted with one selected from the group consisting of ), -cyclohexyl, -cyclobutoxy, -cyclopropoxy, -cyclopentoxy, -F, -Cl, -Br, -I, -CHF2, -CF3, - C1-3alkoxy, -

[0021] NHC(=0)CH3, -C(=0)CH2CH3, -C(=0)N(CH3)2, -C(=0)NH(CH3), -C(=0)NH2, -S(=O)2NHCH3;

[0022] (The above Rai and Ra2 are each independently - H,

[0023] -F, -Cl, -Br, -I or -Ci-5 alkyl, and Yi is one selected from -CH2-, - NH- and - 0-), is replaced,

[0024] Za and Zb are each independently - or one selected from the group consisting of 1) to 18),

[0025] 1) Ci-5 alkyl;

[0026] 2) -F, -Cl, -Br, or -I; i2,

[0027] OWH Im ), (wherein Rb2 is - CH『 , -NH- or - 0-, and Rbs and Rb4 are each independently - H, - NH2, -F, -Cl, -Br, or - 1), pyrimidinyl or pyrrolyl, and RX9 is - H or Ci-5 alkyl;

[0028] 5) -COOH;

[0029] 6) - NRxwRxii, where Rxio or Rxii are each independently -H or - CH3;

[0030] 7) -CFs;

[0031] 8) -CN; morpholinyl or one or more independently Ci-5 alkyl, - F, -C1,

[0032] -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2 substituted morpholinyl; 10) piperidinyl or one or more - H are each independently Ci-5 alkyl, - F, -

[0033] Piperidinyl substituted with Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2;

[0034] 11) piperazinyl or one or more -H7} each independently C1-5 alkyl, - F, -

[0035] Piperazinyl substituted with Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2;

[0036] 12) -0Rxi2, where Rxi2 is -CFs or Ci-3 alkyl;

[0037] 13) -0H;

[0038] 14) , where the number is 0, 1, 2 or 3, and Rxi3 is - NH2, -

[0039] NH(CH₃)₂, -N(CH₃)₂, methylpyrazolyl, pyrazolyl, piperazinyl, piperidinyl, or morpholinyl;

[0040] 15), where RX14 is -CH₂-, -NH-, or -O-, and RX15 is -H, straight-chain or branched C₁₋₅ alkyl, -NH₂, -F, -Cl, -Br, or -I; r' is H

[0041] ..ah..::;:: 3 ",,,,"

[0042] 16) ° 1 Xu—, where RX16 is C₁₋₅ alkyl or -CH₂CH₂N(CH₃)₂;

[0043] 5.. Doubt '••• Doubt

[0044] L J

[0045] 17); or

[0046] 18) - (C₁₋₃ alkyl)-NH₂

[0047] Ak is —CH₂, —(CH₂)₂, —(CH₂)₃— or, —CH₂, —(CH₂)₂, —pH- outside

[0048] (CH₂)₃ - or the -H is each independently unsubstituted or at least one of them is each independently -NH₂, -OH, -NO₂, -C₁₋₅ alkyl, -CH₂NH₂, -CF₃, -OCF₃, -CN, -F,

[0049] -Cl, -Br, or -I substituted;

[0050] L is a linker moiety connecting Ab to} day;

[0051] Ab is an antibody or an antigen-binding fragment thereof; and day is an integer from 1 to 10.

[0052] (2) In the above (1), day is pyridinyl, pyrazolyl, imidazolyl, phenyl, or At this time, the inner side of the above Ri is not substituted or at least one of the inner sides is independently substituted with - NH2, -OH, -N02 or - F,

[0053] R2 is pyridinyl, phenyl, piperidinyl, piperazinyl,

[0054] At this time, each of the R2 is independently unsubstituted or at least one of the R2 is independently - NH2, - C1-5 alkyl, -(C1-5 alkyl) NH2, -morpholinyl,

[0055] -piperazinyl, is replaced,

[0056] Za and Zb are each independently - or one selected from the group consisting of 1) to 6) below,

[0057] 1) -F; i 2 , Rb2 is - CH2 - or - NH-, Rb3 and Rb4 can each independently be -H or - NH2), and RX9 is

[0058] 3) - NRxwRxii, where Rxio and Rxn are both - H;

[0059] 4) Piperidinyl;

[0060] 5) , where Rxi4 is — CH2 — and RX15 is — H or - NH2;

[0061] 、〈S r'1seo

[0062] ...•'.old and young'、、

[0063] 6) ° Sea“® , where RX16 is C1-5 alkyl ,

[0064] Ak can be - CH『 , - (CH2)2 - or - (CH2)3 -.

[0065] (3) In (1) or (2) above, R1 is pyridinyl or pyrrolyl,

[0066] R2 is pyridinyl or phenyl, wherein one of the phenyls is substituted with piperazinyl,

[0067] 0 / \

[0068] >』〔、、사,

[0069] ? V '◊짜

[0070] Za and Zb are each independently -H, -F, -NH2 or - (CH2)c -, where c is 2, (wherein Rb2 is -NH-, and both Rb3 and Rb4 are -H), and RX9 is -CH2-;

[0071] Ak may be -CH2- or - (CH2)2-.

[0072] (4) In any one of the above (1) to (3), the cytotoxic drug moiety represented by the formula III may be a compound of any one of the following Examples 1 to 43:

[0073]

[0074] (5) In any one of the above (1) to (4), the cytotoxic drug moiety represented by the formula III may be a compound of any one of the following formulas

[0075] IV-1 to IV-4: [Formula IV-1]

[0076] (6) In any one of the above (1) to (5), L may include one or more of the following a) to d): a) a cleavable unit; b) a dipeptide, tripeptide or tetrapeptide unit; c) a glucuronide unit; and d) a PEG unit.

[0077] (7) In any one of the above (1) to (6),

[0078] L may be represented by the following chemical formula V:

[0079] [Chemical Formula V] In the above chemical formula V,

[0080] Ly is a single bond, or -valine-citrulline- or -glycine-glycine- phenylalanine-glycine-, and Lx and Ly cannot be single bonds at the same time;

[0081] Lz is a PABC (para-aminobenzyl carbamate), PAB (para-aminobenzyl), or glucuronide moiety; e is an integer from 1 to 6; h and j are each independently an integer from 1 to 3; at least two of f, g, and i are 0, and when f, g, or i are not 0, f or g is an integer from 2 to 16, and i is an integer from 16 to 30.

[0082] (8) In any one of the above (1) to (7),

[0083] L may be represented by any one of the following chemical formulas V-1 to V-10:

[0084] [Chemical Formula V-1]

[0085] [Chemical Formula V-2]

[0086]

[0087] [Chemical Formula V-6]

[0088]

[0089] [Chemical formula v-1

[0090]

[0091] (9) In any one of the above (1) to (8),

[0092] L-D may be represented by any one of the following chemical formulas VI-1 to VI-11:

[0093] [Chemical Formula VI-1]

[0094] [Chemical Formula VI-2]

[0095]

[0096] [Chemical Formula VI-5]

[0097]

[0098] [Chemical Formula VI-8]

[0099]

[0100] (10) In any one of the above (1) to (9), the antibody may target a cancer cell-specific antigen.

[0101] (11) 상기 (10)에 있어서, 암세포 특이적 항원은 5T4, ABL, ABCF1, ACVR1, ACVR1B, ACVR2, ACVR2B, ACVRL1, AD0RA2A, AFP, Aggrecan, AGR2, AICDA, AIF1, AIGI, AKAP1, AKAP2, ALCAM, ALK, AMH, AMHR2, ANGPT1, ANGPT2, ANGPTL3, ANGPTL4, ANPEP, APC, APOCI, AR, 아로마타제 (aromatase), ASPH, ATX, AX1, AXL, AZGP1 (zinc-a- glycoprotein), B4GALNT1, B7, B7.1, B7.2, B7-H1, B7-H3, B7-H4, B7-H6, BAD, BAFF, BAG1, BAI1, BCR, BCL2, BCL6, BCMA, BDNF, BLNK, BLR1 (MDR15), BlyS, BMP1, BMP2, BMP3B (GDFIO) , BMP4, BMP6, BMP8, BMP10, BMPR1A, BMPR1B, BMPR2, BPAG1 (플렉틴), BRCA1, C19orfl0 (IL27w), C3, C4A, C5, C5R1, CA6, CA9, CANT1, CAPRIN-1, CASP1, CASP4, CAV1, CCBP2 (D6 / JAB61), CCL1(1—309), CCLI1(에오탁신), CCL13(MCP-4), CCL15(MIP-Id), CCL16(HCC-4), CCL17(TARC), CCL18(PARC), CCL19(MIP-3b), CCL2(MCP-1), MCAF, CCL20(MIP-3a) , CCL21 (MEP-2), SLC, exodus-2, CCL22(MDC / STC-I), CCL23 (MPIF-I), CCL24 (MPIF- 2 / 에오탁신- 2), CCL25 (TECK), CCL26(에오탁신- 3), CCL27 (CTACK / ILC) ,CCL28, CCL3 (MIP-Ia), CCL4 (MIPIb),CCL5(RANTES) , CCL7 (MCP-3), CCL8 (mcp-2) , CCNA1 , CCNA2, CCND1 , CCNE1 , CCNE2, CCR1 (CKR1 / HM145) , CCR2 (mcp-IRB / RA) , CCR3 (CKR3 / CMKBR3) , CCR4, CCR5(CMKBR5 / ChemR13) , CCR6 (CMKBR6 / CKR-L3 / STRL22 / DRY6) , CCR7 (CKR7 / EBI1), CCR8 또는 CDwl98 (CMKBR8 / TERI / CKR-L1) , CCR9 (GPR-9-6) , CCRL1 (VSHK1), CCRL2 (L-CCR) , CD13, CD164, CD19, CDH6, CDIC, CD2, CD20, CD21, CD200, CD22, CD23, CD24, CD27, CD28, CD29, CD3, CD33, CD35, CD37, CD38, CD3E, CD3G, CD3Z, CD4, CD40, CD40L, CD44, CD45RB, CD47, CD52, CD56, CD69, CD70, CD72, CD74, CD79A, CD79B, CD8, CD80, CD81, CD83, CD86, CD97, CD99, CD117, CD125, CD137, CD147,

[0102] CD179b, CD223 , CD279, CDH1 (E-카드헤린), CDH10, CDH12, CDH13 , CDH18, CDH19, CDH20, CDH3, CDH5, CDH7, CDH8, CDH9, CDH17, CDK2, CDK3, CDK4, CDK5, CDK6,

[0103] CDK7, CDK9, CDKN1A (p21Wapl / Cipl) , CDKN1B (p27Kipl), CDKN1C, CDKN2A (pl6INK4a) , CDKN2B, CDKN2C, CDKN3 , CEA, CEACAM5, CEACAM6, CEBPB, CERI , CFC1B, CHGA, CHGB, 키티나제 (Chitinase), CHST10, CIK, CKLFSF2, CKLFSF3 , CKLFSF4, CKLFSF5, CKLFSF6, CKLFSF7, CKLFSF8, CLDN3 , CLDN6, CLDN7 (클라우딘- 7), CLDN18, CLEC5A, CLEC6A, CLEC11A, CLEC14A, CLN3, CLU (클러스테린), CMKLR1 , CMK0R1 (RDC1), CNR1, C-MET, C0L18A1 , COL I Al, C0L4A3 , C0L6A1 , CR2, Cripto, CRP, CSF1 (M-CSF) , CSF2 (GM-CSF) , CSF3 (GCSF) , CTAG1B (NY-ESO-1), CTLA4, CTL8, CTNNB1 (b-카테닌) , CTSB (카텝신 B), CX3CL1 (SCYD1), CX3CR1 (V28) , CXCL1 (GR01), CXCL10 (IP-IO), CXCLI1 (l-TAC / IP-9), CXCL12 (SDF1), CXCL13 , CXCL14, CXCL16, CXCL2 (GR02) , CXCL3 (GR03), CXCL5 (ENA-78 / LIX) , CXCL6 (GCP-2) , CXCL9 (MIG), CXCR3 (GPR9 / CKR-L2) , CXCR4, CXCR6 (TYMSTR / STRL33 / Bonzo) , CYB5, CYC1, CYSLTR1 , DAB2IP, DES, DKFZp451J0118, DLK1, DNCL1 , DPP4, E2F1, Engel, Edge, Fennel, EFNA3 , EFNB2, EGF, EGFR, ELAC2, ENG, Enola, EN02, EN03, EpCAM, EPHA1 , EPHA2, EPHA3 , EPHA4,EPHA5, EPHA6, EPHA7, EPHA8, EPHA9, EPHA10, EPHB1, EPHB2, EPHB3, EPHB4, EPHB5, EPHB6, EPHRIN-A1, EPHRIN-A2, EPHRINA3, EPHRIN-A4, EPHRIN-A5, EPHRIN-A6, EPHRIN-B1, EPHRIN-B2, EPHRIN-B3, EPHB4, EPG, ERBB2 (HER-2), ERBB3, ERBB4, EREG, ERK8, estrogen receptor (Ear 1 and ESR2), F3 (TF), FADD, FAP, farnesyltransferase, FasL, FASNf, FCER1A, FCER2, FCGR3A, FGF, FGF1 (aFGF), FGF10, FGF11, FGF12, FGF12B, FGF13, FGF14, FGF16, FGF17, FGF18, FGF19, FGF2 (bFGF), FGF20, FGF21, FGF22, FGF23, FGF3 (int-2), FGF4 (HST), FGF5, FGF6 (HST-2), FGF7 (KGF), FGF8, FGF9, FGFR1, FGFR2, FGFR3, FGFR4, FIGF (VEGFD), FILl (EPSILON), FBL1 (ZETA), FL J 12584, FLJ25530, FLRT1 (fibronectin), FLT1,

[0104] , GNRH1 , GPC1, GPC3, GPNB, GPR2 (CCR10), GPR31, GPR44, GPR81 (FKSG80), GRCC10 (CIO), GRP, GSN (Gelsol in), GSTP1 , GUCY2C, HAVCR1 , HAVCR2, HDAC, HDAC4, HDAC5, HDAC7A, HDAC9, Hedgehog, HGF, HIF1A, HIP1, HLA-A, HLA-DR, HLA-DRA, HLA-E, HM74, HMOXI, HSP90, HUMCYT2A, ICEBERG, ICOSL, ID2, IFN-a, IFNA1, IFNA2, IFNA4, IFNA5, EFNA6, BFNA7, IFNB1, IFN감마, IFNW1, IGBP1, IGF1, IGFIR, IGF2, IGFBP2, IGFBP3, IGFBP6, DL-1, ILIO, ILIORA, ILIORB, IL-1, IL1R1 (CD121a), ILlR2(CD121b), IL-IRA, IL-2, IL2RA (CD25) , IL2RB(CD122) , IL2RG(CD132) , IL-4, IL- 4R(CD123) , IL-5, IL5RA(CD125) , IL3RB(CD131) , IL-6, IL6RA, (CD126) , IR6RB(CD130) , IL-7, IL7RA(CD127) , IL-8, CXCR1 (IL8RA), CXCR2, (IL8RB / CD128), IL-9, IL9R(CD129) , IL-10, IL10RA(CD210) , IL10RB(CDW210B) , IL-11, IL11RA, IL-12, IL-12A, IL-12B, IL-12RB1, IL-12RB2, IL-13,IL13RA1, IL13RA2, IL14, IL15, IL15RA, IL16, IL17, IL17A, IL17B, IL17C, IL17R, IL18, IL18BP, IL18R1, IL18RAP, IL19, ILIA, ILIB, ILIF10, ILIF5, IL1F6, ILIF7, IL1F8, DL1F9, ILIHYI, ILIR1, IL1R2, ILIRAP, ILIRAPLI, ILIRAPL2, ILIRL1, IL1RL2, ILIRN, IL2, IL20, IL20RA, IL21R, IL22, IL22R, IL22RA2, IL23, DL24, IL25, IL26, IL27, IL28A, IL28B, IL29, IL2RA, IL2RB, IL2RG, IL3, IL30, IL3RA, IL4, 1L4, IL6ST (Glycoprotein 130), ILK, INHA, INHBA, INSL3, INSL4, IRAKI, IRAK2, ITGA1, ITGA2, ITGA3, ITGA6 (α6 integrin), ITGAV, ITGB3, ITGB4 (β4 integrin), JAG1, JAK1, JAK3, JTB, JUN, K6HF, KAI1, KDR, KIT, KITLG, KLF5 (GC Box BP), KLF6, KLK10, KLK12, KLK13, KLK14, KLK15, KLK3, KLK4, KLK5, KLK6, KLK9, KRT1, KRT19 (Keratin 19), KRT2A, KRTHB6 (Hair-specific type II keratin), L1CAM, LAG3, LAMA5, LAMP1, LEP (Leptin), Lewis Y antigen (LeY), LILRB1, Lingo—p75, Lingo- Troy, LGALS3BP, LRRC15, LPS, LTA (TNF-b), LTB, LTB4R (GPR16), LTB4R2, LTBR, LY75, LYPD3, MACMARCKS, MAG or OMgp, MAGEA3, MAGEA6, MAP2K7 (c-Jun), MDK, MIB1, midkine, MIF,EGYPTIAN, MJP-2, MLSN, MK, MKI67 (Ki-67), MMP2, MMP9, MSMB, MT3 (Enzyme— UI), mTOR, MTSS1 , MUC1 (mucin), MUC16, MYC, MYD88, NCK2, NCR3LG1 , . Neurocan, NFKBI, NFKB2, NGFB (NGF), NGFR, NgR-Lingo, NgRNogo66, (Nogo), NgR— p75, NgR-Troy, NMEI (NM23A), NOTCH, N0TCH1, N0TCH3, N0X5, NPPB, NR0B1 , NR0B2, NRID1, NR1D2, NR1H2, NR1H3 , NR1H4, NR112, NR113, NR2C1 , NR2C2, NR2E1 , NR2E3 , NR2F1 , NR2F2, NR2F6, NR3C1 , NR3C2, NR4A1 , NR4A2, NR4A3 , NR5A1 , NR5A2, NR6A1 , NRP1, NRP2, NT5E, NTN4, NY-ES01 , ODZI, OPRDI , P2RX7, PAP, PARTI , PATE, PAWR, P—가드헤린, PCA3, PCD1, PD-L1 , . PCDGF, PCNA, PDGFA, PDGFB, PDGFRA, PDGFRB, PECAMI , Ll-CAM, peg—proteins, PF4 (CXCL4) , PGF, PGR, phosphacan, PIAS2, PI3 PIK3CG, PLAU(uPA), PLG, PLXDCI, PKC, PKC—PKK, PPBP(CXCL7), PPID, PR1, PRAME, PRKCQ, PRKD1, PRL, PROC, PR0K2, PSAP, PSCA, PSMA, PTAFR, PTEN, PTHR2, PTGS2 (COX— 2), PTN, PVRIG, RAC2 (P21Rac2) , RANK, RANK RARE, RARE, RGS1, RGS13 , RGS3, RNFI10 (ZNF144) , Ron, R0B02, R0R1, RXR, .S100A2, SCGB1D2 (lipophilin B), SCGB2A1 (mammaglobin 2), SCGB2A2 (mammaglobin 1), SCYE1 (endothelial monocyte-activating cytokine), SDF2, SERPENA1, SERPINA3, SERPINB5 (maspin), SERPINEI (PAI-I), SERPINFI, SHIP-1,

[0105] SHIP-2, SHB1, SHB2, SHBG, SfcAZ, SLC2A2, SLC33A1, SLC43A1, SLC44A4, SLC34A2, SLIT2, SPP1, SPRR1B (Sprl), ST6GAL1, ST8SIA1, STAB1, STATE, STEAP, STEAP2, TB4R2, TBX21 , TCP10, TDGF1, TEK, TGFA, TGFB1, TGFB1I1, TGFB2, TGFB3, TGFBI, TGFBR1, TGFBR2, TGFBR3, THIL, THBS1 (Thrombospondin-1), THBS2, THBS4, THPO, TIE (Tie-1), TIMP3, tissue factor (tissue factor) factor), TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11, TNF, TNF-a, TNFAIP2 (B94), TNFAIP3, TNFRSFI1A, TNFRSF1A, TNFRSF1B, TNFRSF21, TNFRSF5, TNFRSF6 (Fas), TNFRSF7, TNFRSF8, TNFRSF9, TNFSF10 (TRAIL), TNFRSF10A, TNFRSF10B, TNFRSF12A, TNFRSF17, TNFSF11 (TRANCE), TNFSF12 (AP03L), TNFSF13 (April), TNFSF13B, TNFSF14 (HVEM-L), TNFRSF14 (HVEM), TNFSF15 (VEGI), TNFSF18, TNFSF4 (CD40 ligand), TNFSF5 (CD40 ligand), TNFSF6 (FasL), TNFSF7 (CD27 ligand), TNFSF8 (CD30 ligand), TNFSF9 (4-1BB ligand), TOLLIP, Toll-like receptor, T0P2A (topoisomerase Ila), TP53, TPM1, TPM2, TRADD, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5, TRAF6, TRKA, TREM1, TREM2, TR0P2, TRPC6, TSLP,TWEAK, 티로시나저] (Tyrosinase), uPAR, VEGF, VEGFB, VEGFC, 베르시칸 (ver si can), VHL C5, VLA-4, WT1, Wnt-1, XCL1 (림포탁틴), XCL2 (SCM-Ib) , XCRI (GPR5 / CCXCR1) , YY1, ZFPM2, CLEC4C (BDCA-2, DLEC, CD303 , CDH6, CLECSF7) , CLEC4D (MCL, CLECSF8) , CLEC4E (Mincle), CLEC6A (덱틴- 2), CLEC5A (MDL-1 , CLECSF5) , CLEC1B (CLEC-2) , CLEC9A (DNGR-1), CLEC7A (덱틴- 1), CLEC11A, PDGFRa, SLAMF7, GP6 (GPVI), LILRA1 (CD85I), LILRA2 (CD85H, ILT1), LILRA4 (CD85G, ILT7) , LILRA5 (CD85F, ILT11), LILRA6 (CD85b, ILT8) , LILRB1, NCR1 (CD335, LY94, NKp46) , NCR3 (CD335, LY94, NKp46) , NCR3 (CD337, NKp30) , OSCAR, TARM1, CD30, CD300C, CD300E, CD300LB (CD300B) , CD300LD (CD300D) ,

[0106] KIR2DL4 (CD158D), KIR2DS, KLRC2 (CD159C, NKG2C), KLRK1 (CD314, NKG2D), NCR2 (CD336, NKp44), PILRB, SIGLEC1 (CD169, SN), SIGLEC5, SIGLEC6, SIGLEC7, SIGLEC8, SIGLEC9, SIGLEC10, SIGLEC11, SIGLEC12, SIGLEC14, SIGLEC15 (CD33L3), SIGLEC16, SIRPA, SIRPB1 (CD172B) , TREM1 (CD354) , TREM2, KLRF1 (NKp80) , 17- 1A, SLAM7, MSLN, CTAG1B / NY-ES0-1, MAGEA3 / A6, ATP5I (Q06185), OAT (P29758), AIFM1 (Q9Z0X1), AGFA (Q64133), MTDC (P18155), CMC1 (Q8BH59), PREP (Q8K411), YMEL1 (088967) . , LPPRC (Q6PB66), LONM (Q8CGK3), ACON (Q99KI0), 0D01 (Q60597), IDHP (P54071), ALDH2 (P47738), ATPB (P56480), AATM (P05202), TMM93 (Q9CQW0), ERG 13 (Q9CQE7), RTN4 (Q99P72), CL041 (Q8BQR4), ERLN2 (Q8BFZ9), TERA (Q01853), DADI (P61804), CALX (P35564), CALU (035887), VAPA (Q9WV55), MOGS(Q80UM7), GANAB(Q8BHN3), ER01A(Q8R180), UGGG1(Q6P5E4), P4HA1(Q60715), HYEP(Q9D379), CALR(P14211), AT2A2(055143), PDIA4(P08003), PDIA1 (P09103), PDIA3 (P27773), PDIA6 (Q922R8), CLH (Q68FD5), PPIB (P24369),TCPG (P80318), M0T4 (P57787),

[0107] NICA (P57716), BAS I (P18572), VAPA (Q9WV55), ENV2 (P11370), VAT1 (Q62465),

[0108] 4F2 (P10852), ENOA (P17182), ILK (055222), GPNMB (Q99P91), ENV1 (P10404),

[0109] ER01A (Q8R180) , CLH (Q68FD5) , DSG1A (Q61495) , AT1A1 (Q8VDN2) , HY0U1 (Q9JKR6) ,

[0110] It may be at least one selected from the group consisting of TRAP1 (Q9CQN1), GRP75 (P38647), ENPL (P08113), CH60 (P63038), and CH10 (Q64433).

[0111] (12) In any one of the above (1) to (11), the antibody may target HER2.

[0112] (13) The pharmaceutical composition for preventing or treating cancer according to the present invention comprises any one of the immunoconjugates (1) to (12) or a pharmaceutically acceptable salt thereof as an active ingredient. (14) The method for preventing or treating cancer according to the present invention comprises a step of administering a therapeutically effective amount of any one of the immunoconjugates (1) to (12) or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0113] (15) One use according to the present invention is the use of any one of the immunoconjugates (1) to (12) or a pharmaceutically acceptable salt thereof for preventing or treating cancer.

[0114] (16) Another use according to the present invention is the use of any one of the immunoconjugates of (1) to (12) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for preventing or treating cancer.

[0115] (17) In any one of (13) to (16), the cancer may be a cancer in which NAMPT is overexpressed or overactivated.

[0116] (18) In any one of (13) to (16), the cancer may be HER2-positive or mutant cancer.

[0117] (19) The drug-linker conjugate according to the present invention or a pharmaceutically acceptable salt thereof is represented by the following Chemical Formula II:

[0118] [Chemical Formula II]

[0119] L-D In Chemical Formula II above, 日 is a cytotoxic drug moiety represented by the following Chemical Formula III,

[0120] [Chemical Formula III]

[0121] In Chemical Formula III above, 日 is pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, pyrrolyl, At this time, the inside of the above Ri is unsubstituted or at least one -

[0122] H7} are each independently substituted with -NH2, -(Ci-3alkyl)NH2, -NH(CI-3alkyl), -N(CI-3alkyl)2, -OH, -NO2, Ci-5alkyl, -F, -Cl, -Br, or -I,

[0123] R2 is pyridinyl, pyrimidinyl, phenyl, piperidinyl, piperazinyl, morpholinyl, pyrazolyl, imidazolyl, phenolic, - CH2C(=0)NH2, - C(=0)NH2, - C(=0)NHCH2CH3, - CH2NHC(=0)CH2CH3, -NH2, -NHCH3, -NHCH2CH2CH3,

[0124] At this time, the inside of the above R2 is independently unsubstituted or at least one of the insides is independently - NH2, - NH (CI-5 alkyl), - MC1-5 alkyl) 2, -NHCHs, -N (CH3) 2, - NHCH2CH3, -OH, - NO2, - S (= 0) 2CH3, - C1-5 alkyl (the inside of the above alkyl is unsubstituted or at least one of the insides is independently substituted with one selected from the group consisting of phenyl, pyridinyl, morpholinyl, piperazinyl and piperidinyl), - (Ci-5 alkyl) NH2, -morpholinyl, -piperidinyl, -piperazinyl, -phenyl, -pyridinyl, -pyrimidinyl (the insides of the above -morpholinyl, - piperidinyl, -piperazinyl, -phenyl, -pyridinyl and -pyrimidinyl are unsubstituted or at least one of the insides is independently - C1-3 alkyl and - CF3 substituted with one selected from the group consisting of ), -cyclohexyl, -cyclobutoxy, -cyclopropoxy, -cyclopentoxy, -F, -Cl, -Br, -I, -CHF2, -CFs, - C1-3alkoxy, - NHC(=0)CH3, -C(=O)CH2CH3, -C(=O)N(CH3)2, -C(=0)NH(CH3), -C(=0)NH2, -S(=O)2NHCH3;

[0125] is replaced by ,

[0126] Za and Zb are each independently - or one selected from the group consisting of 1) to 18),

[0127] 1) Ci-5 alkyl;

[0128] 2) -F, -Cl, -Br, or -I; i2,

[0129] OWH Im ), (wherein Rb2 is - CH『 , -NH- or - 0-, and Rbs and Rb4 are each independently - H, - NH2, -F, -Cl, -Br, or - 1), pyrimidinyl or pyrrolyl, and RX9 is - H, or - C1-5 alkyl;

[0130] 5) -COOH;

[0131] 6) - NRxwRxii, where Rxio or Rxii are each independently -H or - CH3;

[0132] 7) -CFs;Morpholinyl or one or more independently Ci-5 alkyl, -F, -C1,

[0133] -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2 substituted morpholinyl; 10) piperidinyl or one or more - H are each independently Ci-5 alkyl, - F, -

[0134] Piperidinyl substituted with Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2;

[0135] 11) Piperazinyl or one or more -H7} each independently Ci-5 alkyl, - F, -

[0136] Piperazinyl substituted with Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2;

[0137] 12) -0Rxi2, where Rxi2 is -CFs or Ci-3 alkyl;

[0138] 13) -OH;

[0139] 14), wherein q is 0, 1, 2 or 3, and RX13 is -NH2, -

[0140] NH(CH3), -N(CH3)2, methylpyrazolyl, pyrazolyl, piperazinyl, piperidinyl, or morpholinyl;

[0141] 15), wherein RX14 is -CH2-, -NH- or -O-, and RX15 is -H, straight-chain or branched C1-5 alkyl, -NH2, -F, -Cl, -Br, or -I; r 'where H

[0142] ..Ah..::;:: 3 ",,,"

[0143] 16) ° 1 Permit -, wherein RX16 is C1-5 alkyl or -CH2CH2N(CH3)2;

[0144] 5..Fighting '•••Fighting

[0145] L J

[0146] 17); or

[0147] 18) -(C1-3 alkyl)-NH2 Ak is -CH2-, -(CH2)2-, -(CH2)3 - or -(CH2)4,

[0148] (CH2)3 - or are each independently unsubstituted or at least one of

[0149] -Bos are each independently -NH2, -OH, -NO2, -C1-5 alkyl, -CH2NH2, -CF3, -OCF, -CN, -F,

[0150] -Cl, -Br, or -I substituted;

[0151] L is a linker moiety linked to the day.

[0152] (20) In the above (19), the day is pyridine, pyrazole, imidazole, phenyl, or At this time, the inside of the above Ri is unsubstituted or at least one of the insides is independently substituted with -NH2, -OH, -NO2 or -F,

[0153] At least one of the insides is independently -NH2, -C1-5 alkyl, -(C1-5 alkyl)NH2, -morpholine,

[0154] -piperazine, -F or substituted,

[0155] Za and Zb are each independently -visible or one selected from the group consisting of the following 1) to 6),

[0156] 1) -F;

[0157] 2) , where c is 0, 1, 2 or 3, and RX8 is piperidine, i 2, (the above

[0158] Rb2 is -CH2- or -NH-, and Rb3 and blood4 are each independently -H or -NH2),

[0159] RX9 is -H;

[0160] 3) -NRxioRxn, where Rxw and Rxn are both -H;

[0161] 4) Piperidine;

[0162] [[ID=四十二]]

[0162] 5), where RX14 is -CH『and RX15 is -H or -NH2;

[0163] ..,;: :show,,,...

[0164] 6) ° solve “®”, where RX16 is C1-5 alkyl,

[0165] It should be noted that there are some unclear or incorrect expressions in the original text, which may affect the accuracy of the translation. It is recommended to check and correct the original text for a more accurate translation.Ak can be -CH₂, -(CH₂)₂ - or -(CH₂)₃ -

[0166] (21) In the above (19) or (20),

[0167] — 今 r qi

[0168] X우'' f 日 is pyridine or it is,

[0169] R₂ is pyridine or phenyl, where phenyl is substituted with piperazine,

[0170] Za and Zb are each independently -H, -F, where c is 2, (wherein Rb₂ is -NH-, and both Rb₃ and Rb₄ are -H), and RX9 is -H;

[0171] Ak can be -CH₂ - or -(CH₂)₂ -. (22) In any one of the above (19) to (21), the cytotoxic drug moiety represented by the formula III can be a compound of any one of Examples 1 to 43 below:

[0172]

[0173]

[0174] (23) In any one of the above (19) to (22), the cytotoxic drug moiety represented by the formula III can be a compound of any one of the following formulas

[0175] IV-1 to IV-4:

[0176] [Formula IV-1]

[0177] [Formula IV-2]

[0178]

[0179] (24) In any one of the above (19) to (23),

[0180] L may comprise one or more of the following a) to d): a) a cleavable unit; b) a dipeptide, tripeptide or tetrapeptide unit; c) a glucuronide unit; and d) a PEG unit.

[0181] (25) In any one of the above (19) to (24), L may be represented by the following chemical formula V:

[0182] [Chemical Formula V]

[0183] Ly is a single bond, or -valine-citrulline- or -glycine-glycine- phenylalanine-glycine-;

[0184] Lz is a PABC (para-aminobenzyl carbamate), PAB (para-aminobenzyl), or glucuronide moiety; e is an integer from 1 to 6; h and j are each independently an integer from 1 to 3; at least two of f, g, and i are 0, and when f, g, or i are not 0, f or g is an integer from 2 to 16, and i is an integer from 16 to 30.

[0185] (26) In any one of the above (19) to (25),

[0186] L may be represented by any one of the following chemical formulas V-1 to V-10:

[0187] [Chemical Formula V-1] [Chemical Formula V-4]

[0188] [Chemical formula V-8]

[0189] (27) In any one of the above (19) to (26), the drug-linker conjugate or a pharmaceutically acceptable salt thereof may be represented by any one of the following chemical formulas VI-1 to VI-11:

[0190] [Chemical Formula VI-1]

[0191]

[0192] [Chemical Formula VI-4]

[0193]

[0194] [Chemical Formula VI-7]

[0195]

[0196] [Chemical Formula VI-11]

[0197]

[0198] (28) The urea compound according to the present invention, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, or a hydrate thereof, or a solvate thereof, has the following chemical formula:

[0199] It may be indicated as VII:

[0200] [Chemical Formula VII]

[0201]

Effect of the invention

[0202]

Brief description of the drawings

[0203]

Best Mode for Carrying Out the Invention

[0204] [Chemical Formula I]

[0205] Ab-(L-D) p In the above formula, D is a cytotoxic drug moiety;

[0206] L is a linker moiety connecting Ab and D;

[0207] Ab is an antibody or an antigen-binding fragment thereof; and n is an integer of 1 to 10. Another aspect of the present invention is a drug-linker conjugate represented by the following Chemical Formula II or a pharmaceutically acceptable salt thereof.

[0208] [Chemical Formula II]

[0209] L-D In the above formula, D is a cytotoxic drug moiety;

[0210] L is a linker moiety linked to the day. In the present invention, the term "immunoconjugate" means a complex in which a cytotoxic drug-linker conjugate is linked to an antibody or an antigen-binding fragment thereof. In the present invention, the term "drug-linker conjugate" means a substance for the preparation of an immunoconjugate to which an antibody or an antigen-binding fragment thereof is not linked, and can be used as an immunoconjugate by binding to any antibody or an antigen-binding fragment thereof as desired. When the immunoconjugate is administered in vivo, after the antibody or an antigen-binding fragment thereof, which is a component thereof, binds to the antigen targeted, the drug is released so that the drug can act on the target cells and / or surrounding cells, and excellent drug efficacy and reduced side effects can be expected as the target drug. Those skilled in the art can appropriately adjust the DAR (Drug-antibody ratio) of the immunoconjugate according to the form of the linker or the ratio of the antibody and the linker (or drug-linker conjugate) in the preparation of the immunoconjugate, and in the immunoconjugate of the present invention, the DAR can be 1 to 10. As one embodiment, in the immunoconjugate of the present invention, the DAR can be 2 to 10, for example, 2 to 4, 4 to 6, 6 to 8, and specifically can be 2, 4, 6, 8, or 10, but is not limited thereto. The immunoconjugate of the present invention includes an antibody or an antigen-binding fragment thereof, a compound according to any one of (1) to (5) described below as a cytotoxic drug moiety, and at least one of (6) to (12) described below as a linker moiety. In addition, the immunoconjugate of the present invention may be one in which a drug-linker conjugate represented by any one of (13) to (23) described below is linked to an antibody or an antigen-binding fragment thereof. Those skilled in the art can easily recognize and clearly understand from the common technical knowledge that the structure of each moiety may be partially modified as a result of the reaction during the conjugation process. Hereinafter, each component of the immunoconjugate and drug-linker conjugate of the present invention will be described in detail.Cytotoxic drug moiety The present invention provides a compound according to any one of the following (1) to (5), a stereoisomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as a cytotoxic drug moiety.

[0211] (1) A urea compound represented by the following chemical formula III, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof:

[0212] [Chemical Formula III] In the above chemical formula III,

[0213] Ri is pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, pyrrolyl, furanyl, phenyl, phenolic, One - H is independently selected from -NH2, - (C1-3 alkyl) NH2, - NH(Ci-3 alkyl), - MC1-3 alkyl) 2,

[0214] -OH, - NO2, - C1-5 alkyl, - F, -Cl, -Br, or -I may be substituted,

[0215] R2 is pyridinyl, pyrimidinyl, phenyl, piperidinyl, piperazinyl, morpholinyl, pyrazolyl, imidazolyl, phenolic, - CH2C(=0)NH2,

[0216] -C(=0)NH2, -C(=0)NHCH2CH3, -CH2NHC(=0)CH2CH3, -NH2, -NHCH3, -NHCH2CH2CH3, At this time, each of the R2 is independently unsubstituted or at least one of the R2 is independently - NH2, - NH(CI-5 alkyl), - N(CI-5 alkyl) 2, -NHCHs, -N(CH3)2, -NHCH2CH3,

[0217] -0H, - N02, -S(=0)2CHS, - C1-5 alkyl (wherein the alkyl may be unsubstituted or at least one of the inner groups may be independently substituted with one selected from the group consisting of phenyl, pyridinyl, morpholinyl, piperazinyl and piperidinyl), -(C1-5 alkyl) NH2,

[0218] -morpholinyl, -piperidinyl, -piperazinyl, -phenyl, -pyridinyl, -pyrimidinyl (wherein the -morpholinyl, -piperidinyl, -piperazinyl, -phenyl, -pyridinyl and -pyrimidinyl may be unsubstituted or substituted with at least one -H7} independently selected from the group consisting of -Ci-3alkyl and -CF3), -cyclohexyl, cyclobutoxy, -cyclopropoxy, -cyclopentoxy, -F, -Cl, -Br, -I, -CHF2, -CFs,

[0219] - C1-3 alkoxy, -NHC(=O)CHs, -C(=0)CH2CH3, -C(=0)N(CHS)2, -C(=0)NH(CH3), -C(=0)NH2, Each independently - H, -F, -Cl, -Br, -I or - C1-5 alkyl, and Yi can be one selected from -CH2-, -NH-, and -0-, can be replaced with ,

[0220] Zb is independently one selected from the group consisting of - or 1) to 18) below,

[0221] 1)C1-5 alkyl; i2, may be CH2OH), (wherein Rb2 may be -CH2-, -NH- or - 0-, and Rb3 and Rb4 may each independently be - H, - NH2, -F, -Cl, - Br, or - 1), pyrimidinyl or pyrrolyl, and RX9 may be - H or Ci-5 alkyl;

[0222] 5) -COOH;

[0223] 6) -NRxioRxn, where Rxio or Rx n can be independently - H, or - Ofe;

[0224] 7) -CFs;

[0225] 8) -CN;

[0226] 9) Morpholinyl or morpholinyl substituted with one or more of Ci-5 alkyl, - F, -C1, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2, each independently;

[0227] 10) Piperidinyl or piperidinyl in which one or more - H are each independently substituted with Ci-5 alkyl, - F, - Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2;

[0228] 11) Piperazinyl or piperazinyl independently substituted with one or more -H7} Ci-5 alkyl, - F, - Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2;

[0229] 12) -0Rxi2, where Rxi2 can be -CFs or C1-3 alkyl;

[0230] 13) -0H;

[0231] 14) , where the number can be 0, 1, 2 or 3, and Rxi3 is

[0232] -NH2, -NH(CH3), -N(CH3)2, methylpyrazolyl, pyrazolyl, piperazinyl, piperidinyl, or morpholinyl;

[0233] 15), where RX14 can be -CH2-, -NH-, or -O-;

[0234] Rxis can be -H, straight-chain or branched C1-5 alkyl, -NH2, -F, -Cl, -Br, or -I;

[0235] (CH2)3 - or each of which is independently unsubstituted or at least one of each is independently -NH2, -OH, -NO2, -C1-5 alkyl, -CH2NH2, -CF3, -OCF3, -CN, -F,

[0236] -Cl, -Br, or -I and can be substituted.

[0237] (2) In the above (1),

[0238] Ri is pyridinyl, pyrazolyl, imidazolyl, phenyl, and at this time, each of the Ri is unsubstituted or at least one -H can be independently substituted by -NH2, _ OH, -NO2 or -F, and the difference = 1 makes it ~?

[0239] R2 is pyridinyl, phenyl, piperidinyl, piperazinyl, ', independently unsubstituted or at least one of each is independently -NH2, -C1-

[0240] 5 alkyl, - (C1-5 alkyl) NH2, -morpholinyl, -piperazinyl, -F or can be substituted;

[0241] Za and Zb are each independently -visible or one selected from the group consisting of the following 1) to 6),

[0242] 1) -F; i 2 ,

[0243] Rb2 can be - CH2 - or - NH-, and Rb3 and Rb4 are each independently -H or -

[0244] NH2 may be), and Rx9 is mine;

[0245] 3) - NRxwRxii, where Rxio and Rxn are both -;

[0246] 4) Piperidinyl;

[0247] 5) , where RX14 is - CH『, and RX15 can be - H or - NH2; or

[0248] 6) , where RX16 is C1-5 alkyl,

[0249] Ak is -CH2-, - (CH2)2 -, or - (CH2)3 -.

[0250] (3) In (1) or (2) above,

[0251] Ri is pyridinyl or ,

[0252] R2 is pyridinyl or phenyl, wherein one of the phenyl groups is substituted with piperazinyl,

[0253] Za and Zb are each independently - H, -F, -NH2 or Here, c is 2, (The above Rb2 is -NH-, Rbs and Rb4 are both -H), and RX9 is;

[0254] Ak is - CH2 - or - (CH2)2 -.

[0255] (4) In any one of the above (1) to (3), the compound represented by the above chemical formula III may be a compound of Examples 1 to 43 below.

[0256]

[0257]

[0258] (5) In any one of the above (1) to (4), the compound may be a compound represented by the following chemical formulae IV-1 to IV-4.

[0259] [Chemical Formula IV-1]

[0260] 1- (2- ((4-fluorophenyl)ethynyl)- 4'- (piperazin-1-yl)- [l,r-biphenyl] -4- yl)- 3- (2-(pyridin- 3 -yl)ethyl)urea (1— (2— ( (4— f luorophenyl )ethynyl )—4 ' — (piperazin-l-yl )— [1 , 1 ' — biphenyl ]— 4— yl )—3— (2— (pyridin— 3— yl )ethyl )urea)

[0261] [Chemical Formula IV-2]

[0262] 1-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(imidazo[1,2-a]pyridin—7—ylmethyl)urea

[0263] [Chemical Formula IV-3]

[0264] N-(2-(piperazin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-

[0265] (Pyridin-4-yl)phenyl)ethynyl)benzamide (N-(2-(piperazin-l-yl)ethyl)-4-((5-(3-

[0266] (2-(pyr idin-3-yl)ethyl)ur ei do)-2-(pyr idi n-4-y 1)phenyl)ethynyl)benz am ide) [Formula IV-4]

[0267] 1-(3-((4-aminophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(imidazo[l,2—a]pyridin-7-ylmethyl)urea) linker moiety

[0268] L is a linker moiety connecting Ab and any one of the above (1) to (5). The linker moiety of the present invention may include any one or more of the following (6) to (12).

[0269] (6) Cleavable linker or non-cleavable linker

[0270] (7) A linker comprising a cleavable peptide unit such as a dipeptide, tripeptide, or tetrapeptide (e.g., GGFG).

[0271] (8) Glucuronide linker

[0272] (9) Linker containing a PEG group

[0273] (10) Any one linker selected from the group consisting of MP— E(PEG24)— VC— PABC, MP— E(PEG)nVA— PAB, MP— E(PEG)nVK— PAB, MP- E(PEG)nGCit-PAB, MP- K(PEG)n- PEGm- VA- PAB, MP- K(PEG)n- PEGm- VK- PAB, MC- VK(PEG)n- PAB, MC- FK(PEG)n- PAB, MP- K(PEG)nB-VA-PAB (Glue) and MP- K(PEG)n- B- VK- PAB(Gluc).

[0274] (11) A linker represented by the following chemical formula V:

[0275] [Chemical Formula V]

[0276] Ly is a single bond, or -valine-citrulline- or -glycine-glycine- phenylalanine-glycine-, and Lx and Ly cannot be single bonds at the same time;

[0277] Lz is a PABC (para-aminobenzyl carbamate), PAB (para-aminobenzyl), or glucuronide moiety; e is an integer from 1 to 6; h and j are each independently an integer from 1 to 3; at least two of f, g, and i are 0, and when f, g, or sphere is not 0, f or g is an integer from 2 to 16, and sphere is an integer from 16 to 30. For example, in the above formula V, e is an integer from 2 to 5; h and j are 2; at least two of f, g, and i are 0, and when f, g, or sphere is not 0, f or g is an integer from 4 to 12, and sphere can be 24.

[0278] (12) Any one linker selected from the following chemical formulas V-1 to V-10

[0279] [Chemical Formula V-1]

[0280] [Chemical Formula V-2]

[0281]

[0282] [Chemical Formula V-6]

[0283]

[0284] [Chemical formula V-1

[0285] The drug-linker conjugate of the present invention may be a compound represented by any one of the following (13) to (23).

[0286] (13) 4- ( (S)-2- ( (S)-2- (6- (2,5-dioxo-2,5-dihydro- 1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl 4-(2'-((4-fluorophenyl)ethynyl)- 4'- (3- (2-(pyridin-3-yl)ethyl)ureido)- [1,1'-biphenyl]-4-yl)piperazine-1-carboxylate (4- ( (S)-2- ( (S)-2- (6- (2,5-di oxo-2,5-dihydro- 1H- pyrrol-l-yl)hexanam i do ) -3-methy 1 but an am i do ) — 5— ur ei dopent an am i do ) benzy 1 4-

[0287] (2 ' -((4-f luorophenyl )ethynyl )-4 ' - (3- (2- (pyr idin- 3- y 1 )ethyl )ureido)-[l ,1'- biphenyl ]-4-yl )piperazine—l— carboxylate)

[0288] [Chemical Formula VI-1]

[0289]

[0290] (14) 4- ((17S,20S)- 1- (2,5-dioxo-2,5-dihydro-lH-pyrrol-l-yl)-17-isopropyl- 15, 18 -dioxo- 20- (3 -ureidopropyl)- 3, 6, 9, 12 -tetraoxa- 16, 19 - diazahenicosan- 21-amido)benzyl 4- (2'- ((4 -fluorophenyl)ethynyl)- 4'- (3-(2-

[0291] (Pyridin- 3 -yl)ethyl)ureido)- [1,1'-biphenyl] -4 -yl)piperazine-1-carboxylate (4- ((17S,20S)—l— (2,5— dioxo— 2,5— dihydro—lH— pyrrol— 1— y 1)—17— isopropyl— 15, 18 — dioxo— 20— (3— ureidopropyl)— 3,6,9, 12— tetraoxa— 16, 19— diazahenicosan— 21— ami do) benzyl 4-(2 ' — ( (4— f luorophenyl )ethynyl )—4 ' — (3— (2— (pyridin— 3— yl )ethyl )ureido)— [1 , 1 ' -biphenyl ] -4-yl )piperazine—l— carboxylate)

[0292] [Chemical Formula VI-2]

[0293]

[0294] (15) l-(4-((2S, 5S)-37-(2, 5 -dioxo- 2 , 5 -dihydro- 1H-pyrrol- 1-yl)- 5-isopropyl- 4 , 7 , 35 -trioxo- 2- (3 -ureidopropyl )- 10 , 13 , 16 , 19 , 22 , 25 , 28 , 31 -octaoxa- 3 , 6 , 34 -triazaheptatriacontanamido)benzyl )- 4- ( 2- (( 4-fluorophenyl)ethynyl)- 4- (3- (imidazo [ 1 , 2- a]pyridin- 7-ylmethyl)ureido)phenyl)pyridin-1-ium (l-(4-((2S , 5S)-37-(2, 5-dioxo-2, 5- dihydro— 1H— pyrrol— 1—yl )—5— isopropyl— 4,7,35— trioxo— 2—(3— ureidopropyl )- 10,13,16,19,22,25,28, 31— oct aoxa— 3 , 6 , 34-tr i azahept at ri acont anami do) benzyl )-

[0295] 4— (2—( (4— f luorophenyl )ethynyl )—4— (3—( imidazo[l, 2— a]pyr idin— 7— ylmethyl )ureido)phenyl )pyr idin— 1— ium)

[0296] [Chemical Formula VI-3] (16) 4-((2S,5S)-37-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5-isopropyl-4,7,35-trioxo-2-(3-ureidopropyl)-10,13,16,19,22,25,28,31-octaoxa-3,6,34-triazaheptatriacontanamiddo)benzyl 4-(2-(4-((5-(3-(2-

[0297] (pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperazine-1-carboxylate (4-((2S,5S)-37-(2,5-di oxo-2 , 5-di hydr o-lH-pyr rol-l-yl ) -5 - i sopropy 1 — 4 , 7 , 35— tri oxo-2 - (3- ureidopropyl )-10 , 13 , 16, 19,22,25,28, 31— octaoxa— 3 , 6 , 34— tri az aheptat ri acont an am i do ) benzy 1 4— ( 2— ( 4— ( ( 5— ( 3— ( 2— ( pyr idin— 3— yl )ethyl )urei do )-2-( pyr idi n-4-y 1 ) phenyl )ethynyl )benzamido)ethyl )piperazine— 1-carboxylate)

[0298] [Chemical Formula VI-4]

[0299] (17) 4-((2S, 5S)-37-(2, 5-dioxo- 2, 5-dihydro- 1H-pyrrol- 1-yl)- 5-isopropyl- 4, 7, 35-trioxo- 2- (3-ureidopropyl)- 10, 13, 16, 19,22,25,28, 31-octaoxa- 3, 6, 34-triazaheptatriacontanamido)benzyl (4-((5- (3-(imidazo[1, 2-a]pyridin- 7 -ylmethyl)ureido)- 2- (pyridin- 4 -yl)phenyl)ethynyl)phenyl)carbamate (4- ((2S, 5S)— 37— (2, 5— di oxo— 2, 5— di hydro— 1H— pyrrol— 1—yl )— 5— i sopropy 1—4, 7, 35- trioxo- 2-(3- ureidopropyl)- 10,13,16,19,22,25,28,31- octaoxa- 3,6,34- tri azaheptatri acontanamido)benzyl (4— ((5— (3— (imidazo[l,2— a]pyr idin— 7— ylmethyl )ureido)— 2— (pyr idin— 4— yl )phenyl )ethynyl )phenyl )carbamate)

[0300] [Chemical Formula VI-5]

[0301] (18) 4- ((78S,87S)- 87 -Benzyl- 78- (3- (2,5-dioxo-2,5-dihydro-1na-pyrrol-1- yl)propanamido)- 75 ,79,82,85, 88 -pentaoxo-

[0302] 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71- Tetracosaoxa- 74, 80, 83, 86, 89 - Pentaazahennonacontane- 91-amido)benzyl (4- ((5- (3-

[0303] (Imidazo [1,2-a]pyridin-7-ylmethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)phenyl)carbamate (4-((78S,87S)-87-benzyl-78-(3-(2,5-dioxo-

[0304] 2,5— dihydro— 1H— pyrrol— 1—yl )propan amido)— 75,79,82,85,88— pentaoxo—

[0305] 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71- tetracosaoxa-74,80,83,86, 89— pentaazahennonacontan— 91— amido)benzy 1 (4—((5—

[0306] (3— (imidazo[l,2— a]pyridin— 7— ylmethyl)ureido)— 2— (pyridin— 4— yl )phenyl )ethynyl )phenyl )carbamate)

[0307] [Chemical Formula VI-6]

[0308] (19) 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propyl (N-((6S,9S)-1-amino-6-((4-((((4-((5-(3-(imidazo[1,2-a]pyridin-7-ylmethyl)ureido)-2-

[0309] (pyridin-4-yl)phenyl)ethynyl)phenyl)carbamoyl)oxy)methyl)phenyl)carbamoyl)-9-isopropyl-1,8,11-trioxo-14,17,20,23,26,29,32,35-octaoxa-2,7,10-triazaheptatriacontan-37-yl)sulfamoyl)carbamate (3-(2,5-dioxo-2,5-dihydro-

[0310] IH-pyr ro 1 - 1-y 1 ) pr opy 1 (N- ((6S,9S)- 1- amino- 6- ((4- ((((4- ((5- (3- ( imidazo[l,2- a]pyr idin-7-ylmethyl )ureido)— 2— (pyr idin— 4— yl ) phenyl )ethynyl ) pheny 1 ) car b amoy 1 ) oxy ) me t hy 1 )phenyl )carbamoyl )-9 - i sopropy 1—1 ,8, 11-tr i oxo-14 , 17,20,23 ,26,29,32, 35— oct aoxa— 2 , 7 , 10- tri az aheptat ri acont an— 37— y Dsulf amoy 1 ) carbamat e )

[0311] [Chemical Formula VI-7]

[0312] (20) (S)- 1-(4-(5-Benzyl- 55-(2, 5-dioxo- 2, 5-dihydro- 1H-pyrrole- 1-yl)- 4,7, 10, 13, 53 -pentaoxo- 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49 -dodecaoxa-

[0313] 3, 6, 9, 12, 52 -pentaazapentapentacontanamido)benzyl)- 4- (2- ((4-fluorophenyl)ethynyl)- 4- (3-(imidazo [1,2- a]pyridin- 7 -ylmethyl)ureido)phenyl)pyridin- 1-ium ((S)-l-(4- (5— benzyl— 55— (2,5— dioxo— 2,5— dihydro— lH— pyrrol— 1— y 1)— 4, 7, 10, 13, 53— pentaoxo— 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49- dodecaoxa- 3 , 6, 9, 12, 52- pentaazapentapentacontanamido)benzyl )—4— (2—( (4— f luorophenyl )ethynyl )—4—(3— (imidazo[l,2— a]pyridin— 7— ylmethyl)ureido)phenyl)pyridin— 1— ium)

[0314] [Chemical Formula VI-8]

[0315] (21) 1-(4-(((2ne,3chi,4ne,5ne,6ne)-6-carboxy-3,4,5-trihydroxytetrahydro- 2H-pyran- 2 -yl)oxy)- 3- (3- (6- (2, 5 -dioxo- 2,5-dihydro- 1H-pyrrol- 1-yl)hexanamido)propanamido)benzyl)- 4- (2-((4-fluorophenyl)ethynyl)- 4- (3- (imidazo [1,2-a]pyridin- 7-ylmethyl)ureido)phenyl)pyridine— 1—ium (l—(4—((2S,3R,4S,5S,6S)—6—carboxy—

[0316] 3,4,5-tr i hydroxytetr ahydro-2H-pyran-2-y Doxy )-3-(3-(6-(2,5-dioxo-2,5- dihydro—lH— pyrrol— 1—yl )hexanamido)propanamido)benzyl )— 4— (2—((4— f luorophenyl )ethynyl )—4— (3—( imidazo[l ,2— a]pyr idin— 7— ylmethyl )ureido)phenyl )pyr idin— 1— ium)

[0317] [Chemical Formula VI-9]

[0318]

[0319] (22) l-(4-((78S, 81S, 84S)- 78- (3- (2,5 -dioxo- 2,5 -dihydro- 1H-pyrrol- 1-yl)propanamido)- 81 -isopropyl- 75, 79, 82 -trioxo- 84- (3 -ureidopropyl)- 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71- tetracosaoxa- 74, 80, 83 -triazapentaoctacontane- 85 -amido)benzyl )- 4- (2-( (4- Fluorophenyl)ethynyl)-4-(3-(imidazo[1,2-a]pyridin-7-ylmethyl)ureido)phenyl)pyridin-1-ium (1-(4-((78S,81S,84S)-78-(3-(2,5-dioxo-

[0320] 2, 5-dihydro-lH-pyrro 1-1-yl)pr opan ami do)-81- isopropyl -75, 79, 82— tr ioxo— 84— (3— ureidopropyl )- 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59,62,65,68,71- tetracosaoxa-74, 80, 83-tr i azapent aoct acont an-85-ami do) benzyl )— 4— (2— ( (4— f luorophenyl )ethynyl )—4— (3—( imi dazo [ 1 , 2-a ] pyr idin~7~ ylmethyl )ureido)phenyl )pyr idin— 1— ium)

[0321] [Chemical Formula VI-10] (23) (2S,3S,4S,5R,6S)-6-(2-(3-(6-(2, 5 -dioxo- 2, 5 -dihydro- 1H-pyrrol- 1-yl)hexanamido)propanamido)- 4-( ( ( (4-( (5- (3-(imidazo[1,2-a]pyridin- 7-ylmethyl)ureido)- 2-(pyridin- 4-yl)phenyl)ethynyl)phenyl)carbamoyl)oxy)methyl)phenoxy)- 3,4,5-trihydroxytetrahydro- 2H-pyran- 2 -carboxylic acid ( ( 2S, 3S, 4S, 5R, 6S) - 6- ( 2- ( 3- ( 6-

[0322] ( 2 , 5-di oxo-2 , 5-di hydr o-lH-pyr ro 1 — 1— y 1 ) hexanam i do ) propan am i do ) -4 - ((((4—((5—

[0323] (3-( imidazo[l ,2— a]pyr idin— 7— ylmethyl )ureido)-2-(pyr idin-4- yl ) phenyl )ethynyl )phenyl)carb amoy 1 ) oxy ) me t hy 1 ) phenoxy )-3 ,4,5— trihydroxytetrahydro—2H— pyran— 2— carboxyl ic acid)

[0324] [화학식 VI-11] Antibody or antigen-binding fragment thereof A person skilled in the art can prepare an immunoconjugate by attaching an appropriate antibody or antigen-binding fragment thereof to the drug-linker conjugate of the present invention according to the purpose, and the antibody or antigen-binding fragment thereof of the immunoconjugate can recognize a target site and bind thereto to deliver the cytotoxic drug of the present invention into or around a cell. In the present invention, the term "antibody" means a protein molecule that acts as a ligand that specifically recognizes an antigen, including an immunoglobulin molecule that immunologically has reactivity with a specific antigen, and includes polyclonal antibodies, monoclonal antibodies, and whole antibodies. The term also includes chimeric antibodies and bivalent or bispecific molecules, diabodies, triabodies, and tetrabodies. The term further includes single-chain antibodies having a binding function to FcRn, scabs, derivatives of antibody constant regions, and artificial antibodies based on protein scaffolds. A whole antibody has a structure having two full-length light chains and two full-length heavy chains, each light chain being linked to a heavy chain by a disulfide bond. The whole antibody includes IgA, IgD, IgE, IgM, and IgG, and IgG includes subtypes such as IgG1, IgG2, IgG3, and IgG4. In the present invention, the terms "fragment," "antibody fragment," and "antigen-binding fragment" are used interchangeably to refer to any fragment of the antibody of the present invention that retains the antigen-binding function of the antibody. Exemplary antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, Fd, dsFv, and scFv. The Fab has a structure having variable regions of the light chain and heavy chain, constant region of the light chain, and first constant region (CH1 domain) of the heavy chain, and has one antigen-binding site.An antigen-binding fragment of an antibody molecule, or antibody fragment, refers to a fragment that possesses antigen-binding function. Fab' differs from Fab in that it has a hinge region containing one or more cysteine ​​residues at the C-terminus of the heavy chain CH1 domain. F(ab')2 antibodies are generated when cysteine ​​residues in the hinge region of Fab' form disulfide bonds. Dogger refers to the heavy chain portion included in the Fab fragment. Fv (variable fragment) refers to the smallest antibody fragment that contains only the heavy chain variable region and the light chain variable region. Double disulfide Fv (dsFv) has a disulfide bond connecting the heavy chain variable region and the light chain variable region, and single-chain Fv (scFv) can form a dimer-like structure by covalently linking the heavy chain variable region and the single chain variable region through a peptide linker or directly connecting at the C-terminus. Although not limited thereto, such antibody fragments can be obtained using proteolytic enzymes (for example, Fab can be obtained by restriction digestion of the whole antibody with papain, and F(ab')2 fragment can be obtained by digestion with pepsin), or can be produced through genetic recombination technology. In the present invention, but is not limited thereto, the antibody may target a cancer cell-specific antigen. Through this, the immunoconjugate administered in vivo may move to the vicinity of cancer cells, be internalized by cancer cells, and then the drug may be released, or the drug may be released on its own due to the acidic environment around the cancer cells.상기 암 세포 특이적 항원은 예컨대, 5T4, ABL, ABCF1, ACVR1, ACVR1B, ACVR2, ACVR2B, ACVRL1, AD0RA2A, AFP, Aggrecan, AGR2, AICDA, AIF1, AIGI, AKAP1, AKAP2, ALCAM, ALK, AMH, AMHR2, ANGPT1, ANGPT2, ANGPTL3, ANGPTL4, ANPEP, APC, APOCI, AR, 아로마타제 (aromatase), ASPH, ATX, AX1, AXL, AZGP1 (zinc-a- glycoprotein), B4GALNT1, B7, B7.1, B7.2, B7-H1 , B7-H3 , B7-H4, B7-H6, BAD, BAFF, BAG1, BAI1, BCR, BCL2, BCL6, BCMA, BDNF, BLNK, BLR1 (MDR15), BlyS, BMP1, BMP2, BMP3B (GDFIO), BMP4, BMP6, BMP8, BMP10, BMPR1A, BMPR1B, BMPR2, BPAG1 (플렉틴), BRCA1, C19orfl0 (IL27w), C3, C4A, C5, C5R1, CA6, CA9, CANT1,.

[0325] CAPRIN-1, CASP1 , CASP4, CAV1, CCBP2 (D6 / JAB61), CCL1 (1-309), CCLI1 (에오탁신) , CCL13 (MCP-4) , CCL15 (MIP-Id), CCL16 (HCC-4) , CCL17 (TARC) , CCL18 (PARC) , CCL19 (MIP-3b) , CCL2 (MCP-1), MCAF, CCL20 (MIP— 3a), CCL21 (MEP-2), SLC, exodus-2, CCL22(MDC / STC- 1) , CCL23 (MPIF-I), CCL24 (MPIF- 2 / 에오탁신- 2) , CCL25 (TECK) , CCL26(에오탁신- 3) , CCL27 (CTACK / ILC) , CCL28, CCL3 (MIP-Ia), CCL4 (MIPIb), CCL5(RANTES) , CCL7 (MCP-3), CCL8 (mcp-2) , CCNA1 , CCNA2, CCND1 , CCNE1 , CCNE2, CCR1 (CKR1 / HM145) , CCR2 (mcp-IRB / RA) , CCR3 (CKR3 / CMKBR3) , CCR4, CCR5(CMKBR5 / ChemR13) , CCR6 (CMKBR6 / CKR-L3 / STRL22 / DRY6) , CCR7 (CKR7 / EBI1), CCR8 또는 CDwl98 (CMKBR8 / TERI / CKR-L1) , CCR9 (GPR-9-6) , CCRL1 (VSHK1), CCRL2 (L-CCR) , CD13, CD164, CD19, CDH6, CDIC, CD2, CD20, CD21, CD200, CD22, CD23, CD24, CD27, CD28, CD29, CD3, CD33, CD35, CD37, CD38, CD3E, CD3G, CD3Z, CD4, CD40, CD40L, CD44, CD45RB, CD47, CD52, CD56, CD69, CD70, CD72, CD74, CD79A, CD79B, CD8, CD80, CD81, CD83, CD86, CD97, CD99, CD117, CD125, CD137, CD147,CD179b, CD223, CD279, CDH1 (E-카드헤린), CDH10, CDH12, CDH13, CDH18, CDH19, CDH20, CDH3, CDH5, CDH7, CDH8, CDH9, CDH17, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK9, CDKN1A (p21Wapl / Cipl), CDKN1B (p27Kipl), CDKN1C, CDKN2A (pl6INK4a), CDKN2B, CDKN2C, CDKN3, CEA, CEACAM5, CEACAM6, CEBPB, CERI, CFC1B, CHGA, CHGB, 키티나제 (Chitinase), CHST10, CLU (클러스테린), CMKLR1 , CMK0R1 (RDC1), CNR1, C-MET, C0L18A1 , COL I Al, C0L4A3 , C0L6A1 , CR2, Cripto, CRP, CSF1 (M-CSF), CSF2 (GM-CSF), CSF3 (GCSF), CTAG1B (NY-ESO-1) , CTLA4, CTL8, CTNNB1 (b-카테닌), CTSB (카텝신 B), CX3CL1 (SCYD1), CX3CR1 (V28), CXCL1 (GR01), CXCL10 (IP-IO), CXCLI1 (l-TAC / IP-9), CXCL12 (SDF1), CXCL13, CXCL14, CXCL16, CXCL2 (GR02), CXCL3 (GR03), CXCL5 (ENA-78 / LIX), CXCL6 (GCP-2), CXCL9 (MIG), CXCR3 (GPR9 / CKR-L2), CXCR4, CXCR6 (TYMSTR / STRL33 / Bonzo), CYB5, CYC1, CYSLTR1, DAB2IP,DES, DKFZp451J0118, DLK1, DNCL1, DPP4, E2F1, Engel, Edge, Fennel, EFNA3, EFNB2, EGF, EGFR, ELAC2, ENG, Enola, EN02, EN03, EpCAM, EPHA1, EPHA2, EPHA3, EPHA4, EPHA5, EPHA6, EPHA7, EPHA8, EPHA9, EPHA10, EPHB1, EPHB2, EPHB3, EPHB4, EPHB5, EPHB6, EPHRIN-A1, EPHRIN-A2, EPHRINA3, EPHRIN-A4, EPHRIN-A5, EPHRIN-A6, EPHRIN-B1, EPHRIN-B2, EPHRIN-B3, EPHB4, EPG, ERBB2 (HER-2), ERBB3, ERBB4, EREG, ERK8, 에스트로겐 수용체 (Ear 1과 ESR2), F3 (TF), FADD, FAP, FGF10, FGF11, FGF12, FGF12B, FGF13, FGF14, FGF16, FGF17, FGF18, FGF19, FGF2 (bFGF), FGF20, FGF21, FGF22, FGF23, FGF3 (int-2), FGF4 (HST), FGF5, FGF6 (HST-2), FGF7 (KGF), FGF8, FGF9, FGFR1, FGFR2, FGFR3, FGFR4, FIGF (VEGFD), FILl(EPSILON), FBL1 (ZETA), FL J 12584, FLJ25530, FLRT1 (피브로넥틴), FLT1, FLT-3 , FOLR1(FR—알파), FOS, FOSLl(FRA-l), FY (DARC), GABRP (GABAa), GAGEB1, GAGECI, GALNAC4S-6ST, GATA3, GD2, GD3, GDF5, GFI1, GFRA1, GGT1, GM-CSF, GNAS1, GNRH1,GPC1, GPC3, GPNB, GPR2 (CCR10), GPR31, GPR44, GPR81 (FKSG80), GRCC10 (CIO), GRP, GSN (Gelsol in), GSTP1, GUCY2C, HAVCR1, HAVCR2, HDAC, HDAC4, HDAC5, HDAC7A, HDAC9, Hedgehog, HGF, HIF1A, HIP1, histamine and histamine receptor, HLA-,

[0326] A, HLA-DR, HLA-DRA, HLA-E, HM74, HMOXI, HSP90, HUMCYT2A, ICEBERG, ICOSL, ID2, IFN-a, IFNA1, IFNA2, IFNA4, IFNA5, EFNA6, BFNA7, IFNB1, IFN-gamma, IFNW1, IGBP1,

[0327] IGF1, IGFIR, IGF2, IGFBP2, IGFBP3, IGFBP6, DL-1, ILIO, ILIORA, ILIORB, IL-1, IL1R1 (CD121a), ILlR2(CD121b), IL-IRA, IL-2, IL2RA (CD25), IL2RB(CD122), IL2RG(CD132), IL-4, IL-4. 4R(CD123) , IL-5, IL5RA(CD125) , IL3RB(CD131) , IL-6, IL6RA, (CD126) , IR6RB(CD130) , IL-7, IL7RA(CD127) , IL-8, CXCR1 (IL8RA), CXCR2, (IL8RB / CD128), IL-9, IL9R(CD129) , IL-10, IL10RA(CD210) , IL10RB(CDW210B) , IL- 11, IL11RA, IL-12, IL-12A, IL-12B, IL-12RB1, IL-12RB2, IL-13, IL13RA1, IL13RA2, IL14, IL15, IL15RA, IL16, IL17, IL17A, IL17B, IL17C, IL17R, IL18, IL18BP, IL18R1, IL18RAP, IL19, ILIA, ILIB, ILIF10, ILIF5, IL1F6, ILIF7, IL1F8, DL1F9, ILIHYI, ILIR1, IL1R2, ILIRAP, ILIRAPLI , ILIRAPL2, ILIRL1, IL1RL2, ILIRN, IL2, IL20, IL20RA, IL21R, IL22, IL22R, IL22RA2, IL23, DL24, IL25, IL26, IL27, IL28A, IL28B, IL29, IL2RA, IL2RB, IL2RG, IL3, IL30, IL3RA, IL4, 1L4, IL6ST (dangdanbaegjil 130), ILK, INHA, INHBA, INSL3, INSL4, IRAKI, IRAK2, ITGA1. ITGA2, ITGA3, ITGA6 (a6 integrin), ITGAV, ITGB3, ITGB4 (xin4 integrin), JAG1, JAK1, JAK3,JTB, JUN, K6HF, KAI1, KDR, KIT, KITLG, KLF5 (GC Box BP), KLF6, KLK10, KLK12, KLK13 , KLK14, KLK15, KLK3, KLK4, KLK5, KLK6, KLK9, KRT1, KRT19 (keratin 19), KRT2A, KRTHB6 (hair-specific type II keratin), L1CAM, LAG3, LAMA5, LAMP1, LEP (leptin), Lewis Y antigen (LeY), LILRB1, Lingo— p75, Lingo- Troy, LGALS3BP, LRRC15, LPS, LTA (TNF-b), LTB, LTB4R (GPR16), LTB4R2, LTBR, ​​LY75, LYPD3 , MACMARCKS, MAG or OMgp, MAGEA3, MAGEA6, MAP2K7 (c-Jun), MDK, MIB1, midkine, MIF, MISRII, MJP-2, MLSN, MK, MKI67 (Ki-67), MMP2, MMP9, MSMB, MT3 (metalothionectin—UI), mTOR, MTSS1, MUC1 (mucin), MUC16, MYC, MYD88, NCK2, NCR3LG1, neurocan, NFKBI, NFKB2, NGFB (NGF), NGFR, NgR-Lingo,

[0328] NgRNogo66, (Nogo), NgR—p75, NgR-Troy, NMEI (NM23A), NOTCH, N0TCH1, N0TCH3, N0X5, NPPB, NR0B1, NR0B2, NRID1, NR1D2, NR1H2, NR1H3, NR1H4, NR112, NR113, NR2C1, NR2C2, NR2E1, NR2E3, NR2F1, NR2F2, NR2F6, NR3C1, NR3C2, NR4A1, NR4A2, NR4A3, NR5A1, NR5A2, NR6A1, NRP1, NRP2, NT5E, NTN4, NY-ES01 , ODZI, OPRDI, P2RX7, PAP, PARTI, PATE, PAWR, P—gardherin, PCA3, PCD1, PD-L1, PCDGF, PCNA, PDGFA, PDGFB, PDGFRA, PDGFRB, PECAMI, Ll-CAM, peg—heart sparaginager], PF4 (CXCL4), PGF, PGR, phosphacan, PIAS2, PI3 kinase, PIK3CG, PLAU (uPA), PLG, PLXDCI, PKC, PKC—beta, PPBP (CXCL7), PPID, PR1, PRAME, PRKCQ, PRKD1, PRL, PROC, PR0K2, PSAP, PSCA, PSMA, PTAFR, PTEN, PTHR2, PTGS2 (COX—2), PTN, PVRIG, RAC2 (P21Rac2), RANK, RANK ligand, RARE, RGS1, RGS13, RGS3, RNFI10 (ZNF144), Ron, R0B02, R0R1, RXR, S100A2, SCGB1D2 (lipophilin B), SCGB2A1 (mammaglobin 2), SCGB2A2 (mammaglobin 1), SCYE1 (endothelial monocyte-activating cytokine), SDF2, SERPENA1, SERPINA3, SERPINB5 (maspin), SERPINEI (PAI-I), SERPINFI, SHIP-1, SHIP-2, SHB1, SHB2, SHBG, SfcAZ,SLC2A2, SLC33A1, SLC43A1, SLC44A4, SLC34A2, SLIT2, SPP1, SPRR1B (Sprl), ST6GAL1, ST8SIA1, STAB1, STATE, STEAP, STEAP2, TB4R2, TBX21, TCP10, TDGF1, TEK, TGFA, TGFB1 , TGFB1I1 , TGFB2 , TGFB3 , TGFBI , TGFBR1 , TGFBR2 , TGFBR3 , THING , THBS1 (Type 1), THBS2 , THBS4 , THPO , TIE (Tie-1 ), TIMP3 (tissue factor), TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11, TNF, TNF-a, TNFAIP2 (B94) , TNFAIP3, TNFRSFI1A, TNFRSF1A, TNFRSF1B, TNFRSF21 , TNFRSF5, TNFRSF6 (Fas), TNFRSF7, TNFRSF8, TNFRSF9, TNFSF10 (TRAIL), TNFRSF10A, TNFRSF10B, TNFRSF12A, TNFRSF17, TNFSF11 (TRANCE) , TNFSF12 (AP03L) , TNFSF13 (April), TNFSF13B, TNFSF14 (WHO-,

[0329] L), TNFRSF14 (HVEM), TNFSF15 (VEGI), TNFSF18, TNFSF4 (0X40 ligand), TNFSF5 (CD40 ligand), TNFSF6 (FasL), TNFSF7 (CD27 ligand), TNFSF8 (CD30 ligand), TNFSF9 (4-1BB ligand), TOLLIP, Toll-like receptor, T0P2A (topoisomerase Ila), TP53, TPM1, TPM2, TRADD, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5, TRAF6, TRKA, TREM1, TREM2, TR0P2, TRPC6, TSLP, TWEAK, Tyrosinase, uPAR, VEGF, VEGFB, VEGFC, Versican (ver si can), VHL C5, VLA-4, WT1, Wnt-1, XCL1 (lymphotactin), XCL2 (SCM-Ib), , CLEC4E (Mincle), CLEC6A (Dectin-2), CLEC5A (MDL-1, CLECSF5), CLEC1B (CLEC-2), CLEC9A (DNGR-1), CLEC7A (Dectin-1), CLEC11A, PDGFRa, SLAMF7, GP6 (GPVI), LILRA1 (CD85I), LILRA2 (CD85H, ILT1), LILRA4 (CD85G, ILT7), LILRA5 (CD85F, ILT11), LILRA6 (CD85b, ILT8), LILRB1, NCR1 (CD335, LY94, NKp46), NCR3 (CD335, LY94, NKp46), NCR3 (CD337, NKp30), OSCAR, TARM1, CD30, CD300C, CD300E, CD300LB (CD300B), CD300LD (CD300D), KIR2DL4 (CD158D), KIR2DS, KLRC2 (CD159C,NKG2C), KLRK1 (CD314, NKG2D), NCR2 (CD336, NKp44), PILRB, SIGLEC1 (CD169, SN), SIGLEC5, SIGLEC6, SIGLEC7, SIGLEC8, SIGLEC9, SIGLEC10, SIGLEC11, SIGLEC12, SIGLEC14, SIGLEC15 (CD33L3), SIGLEC16, SIRPA, SIRPB1 (CD172B), TREM1 (CD354), TREM2, KLRF1 (NKp80), 17-1A, SLAM7, MSLN, CTAG1B / NY-ES0-1, MAGEA3 / A6, ATP5I (Q06185), OAT (P29758) AIFM1 (Q9Z0X1), AGFA (Q64133), MTDC (P18155), CMC1 (Q8BH59), PREP (Q8K411), YMEL1 (088967), LPPRC (Q6PB66), LONM (Q8CGK3), ACON (Q99KI0), 0D01 (Q60597), IDHP (P54071), ALDH2 (P47738), ATPB (P56480), AATM (P05202), TMM93 (Q9CQW0), ERG I 3 (Q9CQE7), RTN4 (Q99P72), CL041 (Q8BQR4), ERLN2 (Q8BFZ9), TERA (Q01853), DADI (P61804), CALX (P35564), CALU (035887), VAPA (Q9WV55), MOGS (Q80UM7), GANAB (Q8BHN3), ER01A (Q8R180), UGGG1 (Q6P5E4), P4HA1 TCPG (Q60715) (P80318), M0T4 (P57787),

[0330] NICA (P57716), BAS I (P18572), VAPA (Q9WV55), ENV2 (P11370), VAT1 (Q62465),

[0331] 4F2 (P10852), ENOA (P17182), ILK (055222), GPNMB (Q99P91), ENV1 (P10404),

[0332] ER01A (Q8R180), CLH (Q68FD5), DSG1A (Q61495), AT1A1 (Q8VDN2), HY0U1 (Q9JKR6),

[0333] TRAP1 (Q9CQN1), GRP75 (P38647), ENPL (P08113), CH60 (P63038), or CH10 (Q64433), but is not limited thereto. In one embodiment, the antibody or antigen-binding fragment thereof of the present invention may target HER2, and specifically may be trastuzumab, but is not limited thereto. Use of the immunoconjugate or a pharmaceutically acceptable salt thereof and a treatment method using the same Another embodiment of the present invention is a pharmaceutical composition for preventing or treating cancer, comprising the immunoconjugate or a pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutical composition of the present invention exhibits NAMPT inhibitory activity, thereby having an excellent anticancer effect, and is effectively delivered to target cells, so that side effects are rarely observed compared to when a cytotoxic drug is administered alone. In the present invention, the cancer includes all cancers that can be treated by inhibition of NAMPT, and may be solid cancer or blood cancer.For example, pseudomyxoma, intrahepatic cholangiocarcinoma, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, lip cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colon cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla of Vater cancer, bladder cancer, peritoneal cancer, parathyroid cancer, adrenal cancer, paranasal sinus cancer, non-small cell lung cancer, tongue cancer, astrocytoma, small cell lung cancer, pediatric brain cancer, pediatric lymphoma, pediatric leukemia, small intestine cancer, meningioma, esophageal cancer, Selected from the group consisting of glioma, renal pelvic cancer, kidney cancer, heart cancer, duodenal cancer, malignant soft tissue cancer, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, eye cancer, vulvar cancer, ureteral cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, stomach cancer, gastric carcinoid, gastrointestinal stromal cancer, Wilms' cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational trophoblastic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid, vaginal cancer, spinal cancer, acoustic neuroma, pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsillar cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, blood cancer, and thymic cancer It may be one or more types, but is not limited thereto. In addition, the cancer includes not only primary cancer but also metastatic cancer, but is not limited thereto, and the cancer may be a cancer in which NAMPT is overexpressed or overactivated. In addition, it may be a HER2-positive or mutant cancer, such as HER2-positive or mutant breast cancer, gastric cancer, colon cancer, non-small cell lung cancer, but is not limited thereto. In addition, the immunoconjugate of the present invention has an excellent effect even in cancer resistant to ENHERTU® (trastuzumab-deruxtecan), an FDA-approved drug.Therefore, the immunoconjugate of the present invention can exhibit excellent anticancer effects when administered to patients who do not respond to ADC based on HER2 antibody and topoisomerase inhibitor or who have relapsed after treatment with the ADC. In the present invention, the pharmaceutically acceptable salt is a salt commonly used in the pharmaceutical industry, for example, 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, L-ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic 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, It may mean an acid salt prepared from 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, L-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, L-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, L-tartaric acid, thiocyanic acid, toluenesulfonic acid, undecylenic acid or other compounds that can be used as acids, but is not limited thereto. It is not. For administration, the pharmaceutical composition of the present invention may additionally contain at least one type of pharmaceutically acceptable carrier in addition to the immunoconjugate, and may also be used by adding other conventional additives such as antioxidants, buffers, bacteriostatic agents, etc., as needed. In addition, such pharmaceutical composition may be formulated by further adding diluents, dispersants, surfactants, binders, and lubricants thereto.The composition of the present invention may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically) depending on the intended method, wherein the dosage thereof varies within the range depending on the patient's body weight, age, sex, health status, and eating habits, administration time, administration method, excretion rate, disease severity, etc. The daily dosage of the immunoconjugate of the present invention is about 0.001 to 1000 mg / kg and may be administered once a day or in divided doses several times. In addition to the immunoconjugate of the present invention, the pharmaceutical composition of the present invention may further contain as an active ingredient at least one therapeutic agent that exhibits the same or similar medical effect as the immunoconjugate. The therapeutic agent may be selected from the group consisting of a chemotherapeutic agent, a radiotherapy agent, an immunotherapy agent, and a tumor microenvironment modulator. According to one specific embodiment of the present invention, the present invention provides a method for preventing or treating cancer, comprising administering a therapeutically effective amount of the immunoconjugate to a subject in need thereof. The method for preventing or treating cancer according to the present invention includes not only treating the cancer itself before the onset of symptoms, but also suppressing or avoiding such symptoms by administering the immunoconjugate of the present invention. According to one specific embodiment of the present invention, the present invention provides the use of the immunoconjugate of the present invention in the manufacture of a medicament for preventing or treating cancer. To manufacture the medicament, the immunoconjugate of the present invention may be combined with acceptable adjuvants, diluents, carriers, etc., and may be prepared in a complex formulation with other active agents (therapeutic agents) to provide a synergistic effect of the active ingredients. As used herein, "subject" means a mammal, including a human, and "administration" means providing a predetermined substance to a patient by any suitable method. As used herein, the term "therapeutically effective amount" refers to an amount of the immunoconjugate of the present invention that is effective in preventing or treating cancer.The matters mentioned in the uses, compositions, and treatment methods of the present invention are equally applicable unless they are contradictory to each other. The embodiments of the present invention can be modified into many different forms, and the scope of the present invention is not limited to the embodiments described below. In addition, the embodiments of the present invention are provided so that a person having average knowledge in the art can more completely explain the present invention. Furthermore, the expression “comprising” a certain component throughout the specification does not exclude other components unless specifically stated to the contrary, but rather means that other components can be further included.

[0334]

Embodiment of the Invention

[0335] <1. Preparation of cytotoxic drug> Example 1: Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-f luorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea) (Synthetic reaction scheme 1)

[0336] Scheme 1. (a) 4-pyridine boronic acid, Pd[PPh3]4, sodium carbonate (N a2 C03), toluene, ethanol (EtOH), reflux, 1- 3 hours (1-

[0337] 3h); (b) Sodium nitrite (NaN02), KI, HC1, water (water), 0°C 1h (Ih); (c) Pd[PPh3]4,

[0338] Cui, TEA, ACN, room temperature (rt), 1 hour (Ih); (d) Fe, AcOH, 80 °C 2 hours (2h); (e) phenyl chloroformate, pyridine, THF, room temperature (RT),

[0339] 2 hours (2h); (f ) 2— (pyridin— 3— yl)ethan— 1— amine (2— (pyridin— 3— yl)ethan— 1— amine) ,

[0340] TEA, THF, reflux, 24 h (24 h) Step 1(a) Synthesis of 5-nitro-2-(pyridin-4-yl)aniline

[0341] 2-Bromo-5-nitroaniline (3 g, 13.8 mmol, 1.0 eq) and 4-pyridine boronic acid (2.55 g, 1.5 eq) were dissolved in toluene (40 mL) and ethanol (EtOH) (30 mL) to obtain a mixture of Pd[PPh3]4 (3.15 g,

[0342] 0.2 eq) and sodium carbonate (Na2C03) aqueous solution (2M, 16.5 ml, 2.4 eq) in toluene (toluene,

[0343] A mixture of 40 mL of ethanol (EtOH, 30 mL) and 16.5 mL of distilled water was rapidly added to the reaction mixture, which was refluxed for 1-3 hours in a nitrogen atmosphere, cooled to room temperature, and concentrated under reduced pressure. The resulting residue was diluted with distilled water and Toyo, and filtered through Celite. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The resulting residue was purified by column chromatography (silica gel, 30-70% hexane / ethyl acetate) to obtain the title target compound (2.77 g). (Yellow solid, yield 93%) Step 2(b) Synthesis of 4-(2-iodo-4-nitrophenyl)pyridine A solution of sodium nitrite (0.48 g, 1.5 eq) dissolved in water (13 mL) was slowly added dropwise to a suspension of 5-nitro-2-(pyridin-4-yl)aniline (1 g, 4.65 mmol, 1 eq) in concentrated aqueous hydrochloric acid (12 mL) and water (12 mL) at 0°C. The reaction mixture became clear by the end of the addition of the sodium nitrite solution. After the addition, the reaction mixture was stirred at 0°C for 10 min. Then, a solution of potassium iodide (2.31 g, 2.0 eq) dissolved in water (12 mL) was added to the mixture at 0°C. A very viscous reddish-brown mixture was formed, which turned dark brown. The reaction mixture was stirred at room temperature for 1 h, treated with saturated aqueous potassium carbonate solution (pH>8), and extracted with ethyl acetate. The organic layer was washed with water, dried over magnesium sulfate, and concentrated under reduced pressure. The obtained residue was purified by column chromatography (silica gel, hexane / ethyl acetate, 1:1) to give the title target compound (0.84 g, yield 55%). Step 3 (c) 4-(2-((4-fluorophenyl)ethynyl)-4-nitrophenyl)pyridine (4-(2-

[0344] Synthesis of ((4-f luorophenyl )ethynyl )—4— nitrophenyl )pyr idine) Triethylamine (0.1 mL, 3 eq), Cul (1.5 mg, 3 mol%) and Pd[PPh3]4 (9.2 mg,

[0345] 3 mol%) was added to a solution of 4-(2-iodo-4-nitrophenyl)pyridine (87 mg, 0.27 mmol, 1.0 eq) in MeCN (2 mL). The mixture was stirred at room temperature for 5 min under N2 atmosphere. Then, 4-fluorophenyl-acetylene (38 mg, 1.2 eq) was added. The reaction mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The obtained residue was purified by column chromatography (silica gel, hexane / ethyl acetate) to obtain the target compound (82 mg, yield 96.5%). Step 4 (d) Synthesis of 3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)aniline

[0346] A mixture of 4-(2-((4-fluorophenyl)ethynyl)-4-nitrophenyl)pyridine (0.08 g, 0.25 mmol, 1.0 eq), iron (0.14 g, 10 eq), and AcOH (2 mL) was stirred at 80°C for 1-2 h. The catalyst was removed by filtration through Celite, and the filtrate was concentrated under reduced pressure. The obtained black viscous oil residue was basified with aqueous potassium carbonate solution, extracted with ethyl acetate, and the organic layer was concentrated under reduced pressure. The target compound was obtained as a crude product. Step 5 (e) Synthesis of phenyl (3- ((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)carbamate

[0347] 3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)aniline (90 mg, 1.0 eq) and pyridine (40 uL, 2 eq) were dissolved in tetrahydrofuran (2 mL). The reaction solution was cooled with ice, phenyl chloroformate (47 uL, 1.5 eq) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, water (0.1 mL) was added to the reaction mixture, and the reaction mixture was concentrated under reduced pressure. The obtained residue was diluted with water and ethyl acetate. The organic layer was washed with water, dried over magnesium sulfate, and concentrated under reduced pressure. The target compound was obtained as a crude product. Step 6 (f) Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(pyridineyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea A mixture of phenyl (3-((4-fluorophenyl)ethynyl)-4-(pyridineyl)phenyl)carbamate (1.0 eq), 3-(2-aminoethyl)pyridine (3-(2-Aminoethyl)pyridine, 2 eq), and triethylamine (3 eq) in THF (4 mL) was heated at 80 °C for 24 h. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by column chromatography (silica gel, 0-5% dichloromethane / methanol) to obtain the title target compound. P NMR (400 MHz, CDC13) 8.65-8.66 (m, 2H) , 8.44-8.45 (m, 2H) , 7.66 (d, J=2.1 Hz, 1H), 7.60 (dt, J = 7.8, 1.8 Hz, 2H) , 7.54 (m, 2H) , 7.27-7.37 (m, 5H), 6.97-7.02 (m, 2H), 6.94 (s, 1H), 5.08 (t, J = 5.8 Hz, 1H), 3.61 (q, J = 6.4 Hz, 2H), 2.91 (t, J= 6.6 Hz, 2H); MS(ESI) m / z MH. + 437. Example 2: Synthesis of 1-(2-(1H-imidazol-1-yl)ethyl)-3-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)urea (1-(2-(1H-imidazol-1-yl)ethyl)—3— (3—((4— f luorophenyl)ethynyl)—4— (pyridin— 4— yl)phenyl)urea)

[0348] Compound of Example 2 was prepared in substantially the same manner as in Example 1, except that 1H—imidazole-1—ethanamine was used instead of 3-(2-aminoethyl)pyridine in step 6) compared to the synthesis of compound of Example 1. Medium NMR (400 MHz, MeOD) 6 8.59-8.61 (m, 2H), 7.78 (d, J = 2.1 Hz, 1H), 7.74 — 7.66 (m, 3H), 7.36 — 7.48 (m, 4H), 7.19 (m, 1H), 7.10 (m, 2H), 7.00 (m, 1H), 4.20 (t, J = 6.0 Hz, 2H), 3.58 (t, J = 6.0 Hz, 2H); MS(ESI) m / z

[0349] MH + 426. Example 3: 1-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-

[0350] (Imidazole [1,2- a]pyridin- 7 -ylmethyl)urea (1- (3- ((4- f luorophenyl)ethynyl)- 4-

[0351] Synthesis of (pyr idin-4-yl ) phenyl )-3-( imidazo[l,2- a]pyr idiri- 7- ylmethyl ) urea) The compound of Example 3 was prepared in substantially the same manner as in Example 1, except that in step 6), 1-{imidazo[1,2-a]pyridin-7-yl}methanamine dihydrochloride was used instead of 3-(2-aminoethyl)pyridine. 1H NMR (400 MHz, MeOD) δ 8.59-8.61 (m, 2H), 8.40 (d, J = 7.0 Hz, 1H), 7.83 (d, J = 2.2 Hz, 1H), 7.79 (m, 1H), 7.71-7.72 (m, 2H), 7.51 - 7.54 (m, 2H), 7.43 - 7.48 (m, 2H), 7.35 - 7.39 (m, 2H), 7.07 - 7.11 (m, 2H), 6.93 (dd, J = 7.0, 1.6 Hz, 1H), 4.49 (s, 2H); MS(ESI) m / z MH + Example 4: Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(3-hydroxyphenyl)urea (1-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(3-hydroxyphenyl)urea) Compound of Example 4 was prepared in substantially the same manner as in Example 1, except that 3-(2-aminoethyl)phenol hydrobromide was used instead of 3-(2-aminoethyl)pyridine in step 6) compared to the synthesis of compound of Example 1. P NMR (400 MHz, MeOD) 5 8.60 (d, J = 6.1 Hz, 2H) , 7.79 (d, J = 2.2 Hz, 1H), 7.71-7.72 (m, 2H) , 7.36 - 7.46 (m, 4H) , 7.08-7.15 (m, 2H) , 6.70 - 6.75 (m, 2H), 6.65 (dd, J= 8.1, 2.1 Hz, 1H), 3.46 (t, J= 7.1 Hz, 2H), 2.78 (t, J= 7.0 Hz, 2H); MS(ESI) m / z MH + 452. Example 5: Synthesis of 1-((1H-pyrrolo[3,2-c]pyridin-2-yl)methyl)-3-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)urea (1-((lH-pyrrolo[3,2-c]pyr idin-2-yl)methyl)—3— (3—((4-f luorophenyl)ethynyl)-4-(pyr idin-4- yl)phenyl)urea) Compound of Example 5 was prepared in substantially the same manner as in Example 1 except that 1-{1H-pyrrolo[3,2-c]pyridin-2-yl}methanamine dihydrochloride was used instead of 3-(2-aminoethyl)pyridine in step 6) compared to the synthesis of compound of Example 1. P NMR (400 MHz, DMSO) 5 11.45 (s, 1H) , 9.05 (s, 1H) , 8.75 (s, 1H) ,

[0352] 8.65-8.67 (m, 2H) , 8.12 (d, J= 5.7 Hz, 1H) , 7.93 (d, J= 1.9 Hz, 1H) , 7.64—

[0353] 7.68 (m, 2H), 7.44-7.52 (m, 4H), 7.35 (d, J= 5.7 Hz, 1H), 7.24-7.28 (m, 2H),

[0354] 6.85 (t, J = 5.8 Hz, 1H), 6.45 (s, 1H), 4.49 (d, J = 5.7 Hz, 2H); MS(ESI) m / z MH + 462. Example 6: 1-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-

[0355] (2-nitro-1H-imidazol-1-yl)ethyl)urea (1-(3-((4-f luorophenyl )ethynyl )-4-

[0356] Synthesis of (pyr idin-4-yl ) phenyl )- 3- (2- (2- nitro- 1H- imidazol- 1- yl ) ethyl ) urea) Compound of Example 6 was prepared in substantially the same manner as in Example 1, except that 2-(2-nitro-1H-imidazol-1-yl)ethan-1-amine hydrochloride was used instead of 3-(2-aminoethyl)pyridine in step 6) compared to the synthesis of compound of Example 1. (0.044 g, 38%) P NMR (400 MHz, CDCls) 8 8.52 (d, J = 5.4 Hz, 2H) , 8.19 (s, 1H) , 7.67 (s, 1H) , 7.44 (d, J = 5.4 Hz, 2H) , 7.27 (s, 1H) , 7.21 - 7.16 (m, 3H) , 7.09 (s, 1H), 7.02 (s, 1H) , 6.88 (t, J = 8.6 Hz, 2H) , 6.10 (t, J = 5.7 Hz,

[0357] 1H), 4.52 (t, J= 5.7 Hz, 2H), 3.53 (q, J= 5.8 Hz, 2H). MS(ESI) m / z MH + 471 Example 7: Synthesis of 1-(4'-(4,4-difluoropiperidine-1-carbonyl)-2-((4-fluorophenyl)ethynyl)-[1,1'-biphenyl]-4-yl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(4'-(4,4-difluoropiper idine-l-car bony 1)-2-((4-f luorophenyl)ethynyl)—[l,l'— biphenyl] -4-yl)-3-(2-(pyridin-3-yl)ethyl)urea) (Synthetic Reaction Scheme 2)

[0358] Reaction Scheme 2. (a) 4,4-difluoropiperidine hydrochloride, HATU, TEA, DCM, room temperature (RT), 2 h (2 h); (b) 2-bromo-5-nitroaniline, PdC12(dppf), K2CO3, dioxane:water (3: l(v / v)), 90 °C, 2 h (2 h); (c) sodium nitrite (NaN02) f KI, HC1, water (water), 0°C, 1 hour (Ih) ; (d) PdCl2(PPh3)2 f Cui, TEA, ACN, 90 °C 3 h (3h); (e) Fe, AcOH, reflux, 1 h (1h); (f) Phenyl chloroformate, pyridine, THF,

[0359] 0°C—room temperature (RT), 2 h (2 h); (g) 2-(pyridin-3-yl)ethan-l-amine, TEA, THF, 80 °C, 4 h (4 h) Step 1 (a) Synthesis of (4-(4,4-di fluoropiperidine-l-carbonyl)phenyl)boronic acid

[0360] 4-Carboxyphenylboronic acid (0.5 g, 3 mmol, 1.0 eq) was dissolved in DCM (15 ml, 0.2 M), and 4,4-difluoropiperidine hydrochloride (0.57 g, 3.6 mmol, 1.2 eq), HATU (1.48 g, 3.9 mmol, 1.3 eq), and TEA (1.7 ml, 12 mmol) were added. The mixture was stirred at room temperature for 2 hours. Saturated sodium bicarbonate (Sat. NaHC03) aqueous solution was added, and the mixture was extracted with DCM. The organic layer was concentrated under reduced pressure, and the next reaction of step 2) was carried out without separation. Step 2 (b) Synthesis of (2'-amino-4'-nitro-[1,1'-biphenyl]-4-yl)(4,4-difluoropiperidin-1-yl)methanone

[0361] 2-Bromo-5-nitroaniline (2-bromo-5-nitroaniline, 0.5 g, 2.3 mmol, 1.0 eq) was dissolved in 1,4-dioxane:water (3:l(v / v)) (12 ml, 0.2 M), and (4-(4,4-difluoropiperidine—l—carbonyl)phenyl)boronic acid (0.8 g, 3 mmol, 1.3 eq), PdC12(dppf) (0.34 g, 0.46 mmol, 0.2 eq), K2CO3 (0.7 g, 5.06 mmol, 2.2 eq) were added and the mixture was stirred at 90 °C. The mixture was stirred for 2 hours at room temperature and filtered. After cooling to room temperature, water and EA (ethyl acetate) were added to the filtrate to extract the organic layer. The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (1.226 g, 68%). Step 3 (c) (4, 4-difluoropiperidin-1-yl) (2'-iodo-4'-nitro-

[0362] [1, 1'-biphenyl]-4-yl)methanone ((4,4-difluoropiperidin-l-yl)(2'-iodo-4'-nitro-

[0363] Synthesis of [1, 1' -biphenyl ] -4-yl ) methanone )

[0364] (2'-amino-4'-nitro-[l,r-biphenyl]-4-yl)(4,4-difluoropiperidin-1-yl)methanone ((2'-amino-4'-nitro-[1,1'-biphenyl]-4-yl)(4,4-di fluor op i peri din- l-yl)methanone, 0.311 g, 0.861 mmol, 1 eq) was added to 0.9 ml of water and concentrated HC1 (cone. After adding HC1 (0.9 ml), NaN02 (0.059 g, 0.861 mmol, 1 eq) dissolved in 0.9 ml of water and KI (0.143 g, 0.861 mmol, 1 eq) dissolved in 0.9 ml of water were added at 0°C and stirred for 1 hour. Saturated After adding sodium bicarbonate (Sat. NaHC03) aqueous solution and neutralizing, it was extracted with ethanol. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.089 g, 22%). Step 4 (d) Synthesis of (4, 4-difluoropiperidin-1-yl)(2'-((4-fluorophenyl)ethynyl)-4'-nitro-[1,1'-biphenyl]-4-yl)methanone ((4,4- dif luoropiperidin-1-yl)(2'-((4- f luorophenyl)ethynyl)-4'-nitro-[1, 1'-biphenyl]-4-yl )methanone)

[0365] (4, 4-Difluoropiperidin-1-yl)(2'-iodo-4'-nitro-[1,1'-biphenyl]-4- yl)methanone ((4,4- dif luoropiperidin- l-yl)(2'-iodo-4'-nitro-[1,1'-biphenyl]- 4- yDmethanone, 0.089 g, 0.188 mmol, 1 eq) was dissolved in ACN (acetonitrile, 2 ml, 0.1 M), then TEA (triethylamine, 0.06 ml, 0.414 mmol, 2.2 eq), PdC12(PPh3)2 (4 mg, 0.006 mmol, 3 mol%), and Cui (2 mg, 0.008 mmol, 4 mol%) were added and stirred at room temperature for 5 min. 1-Ethynyl-4-f luorobenzene (1-ethynyl-4-f luorobenzene, 0.032 ml, 0.283 mmol, 1.5 eq) was added and stirred at 90°C for 3 hours. After cooling to room temperature, the mixture was concentrated and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.075 g, 86%). Step 5 (e) (4'-Amino-2'-((4-fluorophenyl)ethynyl)-[1,1'-biphenyl]-4-yl)(4, 4-difluoropiperidin-1-yl)methanone ((4'-amino-2'-((4-f luorophenyl )ethynyl )-[l, 1' -biphenyl ]— 4— y 1 ) (4, 4-di f 1 uor op i Synthesis of per idi n- 1- yl )methanone)

[0366] (4, 4 -difluoropiperidin- 1-yl) (2 '-((4 -fluorophenyl)ethynyl)- 4'-nitro- [1,1'-biphenyl] -4 -yl)methanone ((4,4- di fluor op i peri din- 1-yl )(2'- ((4- f luorophenyl )ethynyl )— 4 1 — ni tro— [ 1 , 1 1-biphenyl ] -4-yl ) methanone , 0.075 g, 0.161 mmol, 1 eq) was dissolved in AcOH (acetic acid, 1.3 ml, 0.13 MH], Fe (0.090 g, 1.61 mmol, 10 eq) was added, and refluxed for 1 hour. After cooling to room temperature, the mixture was filtered through a Celite filter, water was added to the filtrate, and extracted with DCM (dichloromethane). The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.030 g, 43%). Step 6 (f) Phenyl (4'- (4, 4-difluoropiperidine-1-carbonyl)- 2- ((4- Synthesis of phenyl (4'-(4,4-difluorophenyl)ethynyl)-[1,1'-biphenyl]-4-yl)carbamate (phenyl (4'-(4,4-difluorophenyl)ethynyl)-2-((4-f 1 uor opheny 1 )ethynyl )—[l,l'—biphenyl]—4-yl )carbamate)

[0367] (4'-amino-2'-((4-fluorophenyl)ethynyl)-[1,T-biphenyl]-4-yl)(4,4-difluoropiperidin-1-yl)methanone ( (4 1 -ami no-2 1- ((4- f luorophenyl )ethynyl )- [1,1'— biphenyl]— 4— yl)(4, 4— difluoropiperidin—l—yl)methanone, 0.030 g, 0.069 mmol, 1 eq) was dissolved in THF (tetrahydrofuran, 1 ml, 0.1 ⑴), and pyridine (pyridine, 0.011 ml, 0.138 mmol, 2 eq) and phenyl chloroformate (Phenyl chloroformate, 0.013 ml, 0.104 mmol, 1.5 eq) were added at 0°C, and stirred at room temperature for 2 hours. 0.1 ml of water was added, stirred for 10 minutes, concentrated, and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.027 g, 71 %) was obtained. Step 7 (g) 1- (4'- (4, 4-difluoropiperidine- 1-carbonyl)- 2- ((4-

[0368] ■Synthesis of phenyl (4'-(4,4-difluoropiperidine-1-carbonyl)- 2-((4-fluorophenyl)ethynyl)- [1,1'-biphenyl] -4 -yl)- 3- (2-(pyridin- 3 -yl)ethyl)urea) (1- (4'-(4,4-difluoropiperidine-1-carbonyl)- 2-((4-fluorophenyl)ethynyl)- [1,1'-biphenyl] -4 -yl)carbamate (phenyl (4'-(4,4~ di f luoropiper idine-l-car bonyl )— 2—((4— f luorophenyl )ethynyl )-[l, 1 1-biphenyl]- 4-yl) carbamate, 0.027 g, 0.049 mmol, 1 eq) was dissolved in THF (1 ml, 0.06 M), and 2-(pyridin-3-yl)ethan-1-amine (2-(pyridin-3- yl)ethan-1-amine, 0.011 ml, 0.097 mmol, 2 eq) and TEA (0.02 ml, 0.147 mmol, 3 eq) were added and stirred at 80°C for 4 h. The reaction mixture was concentrated and purified by column chromatography (silica gel, MC / MeOH) to obtain the title compound (0.020 g, 70%). P NMR (400 MHz, MeOD) 5 8.47 (s, 1H) , 8.41 (d, J = 4.8 Hz, 1H) , 7.80 (d, J = 7.8 Hz, 1H) , 7.74 (s, 1H) , 7.72 (s, 2H) , 7.57 - 7.52 (m, 2H) , 7.44 - 7.39 (m, 2H) , 7.38 - 7.32 (m, 3H) , 7.08 (t, J = 8.7 Hz, 2H) , 3.98 - 3.57 (m, 4H), 3.50 (t, J = 7.0 Hz, 2H), 2.92 (t, J = 6.9 Hz, 2H), 2.07 (s,

[0369] 4H). MS(ESI) m / z MH + 583 Example 8: Synthesis of N-(2-(piperidin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (Synthetic Reaction Scheme 3)

[0370] Scheme 3. Reagents and conditions: (a) 2-(piperidin-1-yl)ethan-1-amine, HATU, DIPEA, DMF, room temperature (rt), overnight; (b) NaN02, KI, HC1, H20, 5 °C, 1 h; (c) PdC12(PPh3)2, Cui, TEA, ACN, 60 °C, 5 h; (d) Zn, NH4CI, 1,4-dioxane, water (H2O), room temperature (rt), overnight; (e) phenyl chloroformate, pyridine, THF, at 0 °C Room temperature (rt), 2 h (2 h); (f ) 2— (pyridin— 3— yl)ethan— 1— amine, TEA, THF, room temperature (rt), overnight Step 1 (a) Synthesis of 4-ethynyl- N- (2- (piperidin- 1- yl) ethyl) benzamide

[0371] 4—Ethynyl benzoic acid (0.5 g, 3.42 mmol, 1 eq) was dissolved in DMF (11 ml, 0.3 M), and 2—(piperidin—l— yl)ethan—l— amine (0.657 g, 5.13 eq, 1.5 eq), HATU (1.950 g, 5.13 mmol, 1.5 eq), and DIPEA (1.326 g, 10.26 mmol, 3 eq) were added and stirred at room temperature overnight. The organic layer was extracted with doyu and 0, dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.788 g, 89.7%). Step 2 (b) Synthesis of 4-(2-iodo-4-nitrophenyl)pyridine

[0372] 5-Nitro-2-(pyridin-4-yl)aniline (5-nitro-2-(pyridin-4-yl)aniline, 1.00 g, 4.64 mmol, 1 eq) was added to 4.5 ml of water and 4.5 ml of concentrated HC 1 (cone. HC1), and then NaN02 (0.326 g, 4.73 mmol, 1.0 eq) dissolved in 4.5 ml of water was slowly added dropwise while stirring at 5 °C. After the addition, the mixture was stirred for 10 minutes, and KI (1.464 g, 8.82 mmol, 2 eq) dissolved in 4.5 ml of water was added, and the temperature was raised to room temperature and stirred for 30 minutes. Saturated K2CO3 (Sat. K2CO3) aqueous solution was added, neutralized to pH 8 or higher, and extracted with DCM. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, EA / Hex) to obtain the title compound (0.952 g, 62.9%). Step 3 (c) 4- ((5-nitro-2- (pyridin- 4-yl)phenyl)ethynyl)- N- (2- (piperidin- 1-yl)ethyl)benzamide (4- ((5-nitro-2- (pyridin- 4- yl)phenyl)ethynyl)- N- (2-

[0373] Synthesis of 4-(2-iodo-4-nitrophenyl)pyridine (4-(2-iodo-4-nitrophenyl)pyridine, (piper idin-l-yl )ethyl )benzamide)

[0374] 0.5 g, 1.53 mmol, 1 eq), 4-ethynyl- N- (2-(piperidin- 1-yl)ethyl)benzamide (4-ethynyl— N— (2— (piperidin— 1— yl)ethyl)benzamide, 0.588 g, 2.295 mmol , 1.5 eq), PdC12(PPh3)2 (0.0322 g, 0.046 mmol, 0.03 eq), Cui (0.00876 g, 0.046 mmol, 0.03 eq) were added and degassed with nitrogen gas. After that, ACN (3.06 ml, 0.5 M) and TEA (0.309 g, 3.06 mmol, 2 eq) were added, heated to 60 °C, and stirred for 5 hours. After completion of the reaction, extraction was performed with EA, and the organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.29 g, 41.7%). Step 4 (d) Synthesis of 4-((5-amino-2-(pyridin-4-yl)phenyl)ethynyl)-N-(2-(piperidin-1-yl)ethyl)benzamide (4-((5-amino-2-(pyridin-4-yl)phenyl)ethynyl)-N-(2-(piperidin-1-yl)ethyl)benzamide)

[0375] 4- ((5-nitro-2-(pyridin-4-yl)phenyl)ethynyl)- N- (2-(piperidin-1-yl)ethyl)benzamide (4- ( (5-nitro-2-(pyridin-4- y1)phenyl)ethynyl)- N- (2-

[0376] (piper idin-l-yl )ethyl )benzamide, 0.29 g, 0.64 mmol , 1 eq) was dissolved in 1,4-dioxane (1,4-dioxane): water OfeO) (3:l(v / v)) (1.5 ml, 0.4 M), then Zn (0.418 g, 6.4 mmol , 10 eq) and NH4CI (0.342 g, 6.4 mmol , 10 eq) were added and stirred at room temperature overnight. The mixture was washed with MeOH and filtered under reduced pressure. The filtrate was concentrated and used for the next reaction in step 5) without purification. Step 5 (e) Synthesis of phenyl (3-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)-4-(pyridin-4-yl)phenyl)carbamate

[0377] 4— ( (5—amino—2_(pyridin— 4 -yl)phenyl)ethynyl)- N- (2-(piperidin- 1-yl)ethyl )benzamide (4- ( (5-amino- 2- (pyridin- 4- y 1 )phenyl )ethynyl )- N- (2-

[0378] (piper i di nl-yl )ethyl )benzamide , 0.272 g, 0.64 mmol , 1 eq) was dissolved in THF (3 ml , 0.2 M), and then pyridine (pyr idine , 0.063 g, 0.8 mmol , 1.25 eq) was added.

[0379] After cooling to 0 °C, phenyl chloroformate (0.180 g, 1.152 mmol, 1.8 eq) was added dropwise. The mixture was slowly heated to room temperature and stirred for 2 hours. After completion of the reaction, the mixture was concentrated, neutralized with 1 N NaOH, and extracted with EA and water (Compound 0). The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.115 g, 31.9%). Step 6 (f) Synthesis of N-(2-(piperidin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (N-(2-(piperidin-l-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide) Phenyl (3-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)-4-(pyridin-

[0380] 4-(4-Phenyl)phenyl (3-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)-4-(pyridin-4-yl)phenyl)carbamate, 0.115 g, 0.21 mmol, 1 eq) was dissolved in THF (2 ml, 0.1 M), then 2-(pyridin-3-yl)ethan-1-amine (0.0513 g, 0.42 mmol, 2 eq), TEA (0.064 g, 0.63 mmol, 3 eq) were added and stirred at room temperature overnight. After completion of the reaction, it was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain the title compound (0.016 g, 13.7%). 1H NMR (400 MHz, DMSO) δ 8.91 (s, 1H), 8.68 (s, 2H), 8.57 - 8.42 (m,

[0381] 3H), 8.20 (s, 1H), 7.94 (s, 1H), 7.86 (d, J = 8.0 Hz, 2H), 7.72 - 7.63 (m, 3H), 7.53 - 7.43 (m, 4H), 7.39 - 7.33 (m, 1H), 6.42 (s, 1H), 3.41 (m, 4H), 2.82 (t, J = 6.6 Hz, 2H), 2.55 (m, 5H), 1.54 (m, 4H), 1.41 (m, 2H); MS(ESI) m / z MH+ 573.1 Example 9: Synthesis of 1-(2-((4-fluorophenyl)ethynyl)-3'-morpholino-[1,1'-biphenyl]-4-yl)-3-(2-(pyridin-3-yl)ethyl)urea

[0382] (Synthetic reaction scheme 4)

[0383] Scheme 4. Reagents and conditions: (a) l-ethynyl-4-f luorobenzene, TEA, Cui, PdC12(PPh3)2, ACN, reflux (reflux), 1 h (Ih); (b) Phenyl chloroformate, pyridine, THF, 0°C, 2 h (2h); (c) 2-(pyridin-3-yl)ethan-1-amine, TEA, THF, 80°C, 2 h (2 h): (d) Fe, AcOH, reflux (reflux), 1 h (Ih); (e) NaN02, KI, pTSA, ACN, 5 °C, 1 h (1 h); (f) (3-morpholi nophenyl)boronic acid, PdCLXjdppf , K2CO3, 1,4-di oxane, H2O, 90 °C, 4 h (4 h) Step 1 (a) Synthesis of 3-((4-fluorophenyl)ethynyl)-4-nitroaniline

[0384] 3-Bromo-4-nitroaniline (2.17 g, 0.01 mol, 1 eq) was dissolved in ACN (100 ml, 0.1 M), and TEA (3 ml, 0.22 mol, 2.2 eq), PdC12(PPh3)2 (210 mg, 0.3 mmol, 3 mol%), and Cui (80 mg, 0.4 mmol, 4 mol%) were added. The mixture was stirred at room temperature for 5 min. 1-Ethynyl-4-f luorobenzene (1.72 ml, 0.015 mmol, 1.5 eq) was added and stirred at 90°C for 1 h. After cooling to room temperature, concentration was achieved and the residue was purified by column chromatography (silica gel, EA / HX) to obtain the title compound (1.44 g, 56%). Step 2(b) Synthesis of phenyl (3-((4-'sulfophenyl)ethynyl)-4-nitrophenyl)carbamate (phenyl (3-((4-f fluorophenyl)ethynyl)-4-nitrophenyl)carbamate)

[0385] 3-((4-f luorophenyl)ethynyl)-4-nitroaniline (3-((4- f luorophenyl)ethynyl )—4— nitroaniline, 1.436 g, 5.60 mmol , 1 eq) was dissolved in THF (56 ml, 0.1 M), and pyridine (0.9 ml, 11.2 mmol , 2 eq) and phenyl chloroformate (1.06 ml, 8.41 mmol , 1.5 eq) were added dropwise at 0°C, and stirred at room temperature for 2 hours. Water was added, stirred for 10 minutes, concentrated, and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (1.968 g,

[0386] 93.4%) was obtained. Step 3 (c) 1-(3-((4-fluorophenyl)ethynyl)-4-nitrophenyl)-3-(2-(pyridin-3-yl)ethyl)urea 0}(1—(3—((4—fluorophenyl)ethynyl)—4—nitrophenyl)—3—(2—

[0387] (pyridin-3-yl)ethyl)urea) Synthesis Phenyl (3-((4-fluorophenyl)ethynyl)-4-nitrophenyl)carbamate (phenyl (3-((4-fluorophenyl)ethynyl)-4~nitrophenyl)carbamate, 1 g, 2.66 mmol, 1 eq) was dissolved in THF (1 ml, 0.06 M), then 2-(pyridin-3-yl)ethan-1-amine (2-(hex7heart(1辻—3—yl)ethan—1—amine, 0.62 ml, 5.32 mmol, 2 eq), TEA (1.11 ml, 7.98 mmol, 3 eq) were added and stirred at 80 °C for 4 hours. The reaction mixture was concentrated and purified by column chromatography (silica gel, MC / MeOH) to obtain the title compound (0.954 g, 96%). Step 4 (d) 1-(4-amino-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(4-amino-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-

[0388] (pyridin-3-yl)ethyl)urea) Synthesis

[0389] 1-(3-((4-fluorophenyl)ethynyl)-4-nitrophenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-fluorophenyl)ethynyl)-4-nitrophenyl)-3-(2 -

[0390] (pyridin-3-yl)ethyl)urea (0.954 g, 2.55 mmol, 1 eq) was dissolved in AcOH (20 ml, 0.13 M), and Fe (1.42 g, 25.5 mmol, 10 eq) was added and refluxed for 1 hour. After cooling to room temperature, the mixture was filtered through a Celite filter, water was added to the filtrate, and extracted with DCM. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.859 g, 90%). Medium NMR (400 MHz, DMSO) 6 8.46 (d, J = 1.5 Hz, 1H), 8.43 (d, J = 3.9 Hz, 1H), 8.06 (s, 1H), 7.39 - 7.31 (m, 2H) , 7.26 (t, J = 8.8 Hz, 2H) , 6.98

[0391] (dd, J = 8.7, 2.4 Hz, 1H) , 6.64 (d, J = 8.7 Hz, 1H) , 6.06 - 5.99 (m, 1H) , 5.18 (s, 2H) , 3.36 - 3.30 (m, 2H) , 2.77 (t, J = 7.0 Hz, 2H) . Step 5 (e) Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-iodophenyl)-3-(2-(pyridin- 3 -yl)ethyl)urea

[0392] 1-(4-amino-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea ( 1-(4-amino-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea ( 1-(4-amino-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea

[0393] 3-yl )ethyl )urea, 0.05 g, 0.134 mmol , 1 eq) was dissolved in ACN (1.34 ml, 0.1 M), pTSA (p-toluenesulfonic acid, 0.069 g, 0.402 mmol, 3 eq) was added, and then NaN02 (0.01 g, 0.136 mmol, 1.02 eq) dissolved in 0.15 ml of water at 0°C and KI (0.045 g, 0.268 mmol, 2 eq) dissolved in 0.15 ml of water were added. After reacting at room temperature for 1 hour, saturated sodium bicarbonate (Sat. NaHCOs) aqueous solution was added, neutralized to pH 8 or higher, and extracted with a toyo. The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.038 g, 58.4%). Step 6 (f) 1-(2-((4-fluorophenyl)ethynyl)-3'-morpholino-[1,1'-biphenyl]-

[0394] Synthesis of 1-(2-((4-fl uor ophenyl )ethynyl )-3 '- mor pho 1 ino-[l, 1 '-biphenyl ]-4-yl )-3-(2-(pyridin- 3-yl )ethyl )urea)

[0395] 1-(3-((4-Fluorophenyl)ethynyl)-4-iodophenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-fluorophenyl)ethynyl)-4-iodophenyl)-3-(2-(pyridin-3-yl)ethyl)urea, 0.05 g, 0.1 mmol, 1 eq) was dissolved in 1,4-dioxane: water (H2O) (3:1 (V / V)) (0.5 ml, 0.2 M). After that, (3-morpholinophenyl)boronic acid (0.041 g, 0.2 mmol, 2 eq), PdCl2(dppf) (0.0146 g, 0.02 mmol, 0.2 eq), and K2CO3 (0.0304 g, 0.22 mmol, 2.2 eq) were added, and the mixture was heated to 90 °C and stirred for 4 hours. After completion of the reaction, it was washed with toluene and filtered through a celite filter, and then extracted with EA and H2O. The organic layer was dried over MgSO4, filtered, concentrated, and filtered through prep-LC (0.1% formic acid in H2O / ACN) to obtain the title compound (0.078 g, 15.0%). 1H NMR (400 MHz, DMSO) δ 8.74 (s, 1H), 8.48 (s, 1H), 8.45 (d, J = 4.4

[0396] Hz, 1H), 7.84 (m, 1H), 7.69 (m, 1H), 7.42 (dd, J = 8.3, 5.6 Hz, 2H), 7.37

[0397] (s, 2H), 7.33 (m, 2H), 7.26 (t, J = 8.8 Hz, 2H), 7.18 (s, 1H), 7.04 (d, J =

[0398] 7.4 Hz, 1H), 6.96 (d, J = 8.6 Hz, 1H), 6.30 (t, J = 5.5 Hz, 1H), 3.74 - 3.70

[0399] (m, 4H), 3.39 (dd, J = 12.9, 6.6 Hz, 2H) , 3.15 - 3.10 (m, 4H) , 2.81 (t, J = 6.9 Hz, 2H).; MS(ESI) m / z MH— 519.48 Example 10: 1- (2- ((4-fluorophenyl)ethynyl)- 4'- (piperazin-1-yl)- [1,1'-biphenyl] -4 -yl)- 3- (2-(pyridin-3 -yl)ethyl)urea (1- (2-((4-fluorophenyl)ethynyl)— 4'— (piper azin— 1—yl)-[l, 1' -biphenyl ] -4-yl )-3- (2 -

[0400] Synthesis of (pyr idin-3-yl)ethyl)urea (Synthetic reaction scheme 5)

[0401] Scheme 5. (a) PdCMdppf), K2CO3, 1,4-dioxane (1,4-dioxane), water (H2O), 90 °C, 4 h (4 h) : (b) TFA, DCM, room temperature (rt), overnight. Step 1 (a) tert-Butyl 4-(2'-((4-fluorophenyl)ethynyl)- 4'-(3-(2-

[0402] Synthesis of tert-butyl 4-(2'-((4-f luorophenyl )ethyny 1 )-4'-(3-(2-(pyridin-3- yl)ethyl)ureido)- [1,1'-biphenyl]- 4-yl)piperazine-1-carboxylate

[0403] 1-(3-((4-fluorophenyl)ethynyl)-4-iodophenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-fluor ophenyl)ethyny 1)-4-iodophenyl)-3-(2-(pyri din-3-yl )ethyl )urea, 0.05 g, 0.1 mmol , 1 eq), was dissolved in 1,4-dioxane:water (H20)(3:1(V / V)) (0.5 ml, 0.2 M) and then (4-(4-(tert-butoxycarbonyl)piperazin— 1—yl )phenyl )boronic acid, After adding 0.061 g, 0.2 mmol, 2 eq), PdC12(dppf) (0.0146 g, 0.02 mmol, 0.2 eq), K2C0s (0.0304 g, 0.22 mmol, 2.2 eq), the mixture was heated to 90°C and stirred for 4 hours. After completion of the reaction, the mixture was washed with a toaster, filtered through a celite filter, and extracted with EA, Seong0. The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.043 g, 69.4%). Step 2 (b) 1- (2- ((4-fluorophenyl)ethynyl)- 4'- (piperazin-1-yl)- [1,1'-biphenyl]-4 -yl)- Synthesis of 3-(2-(pyridin- 3 -yl)ethyl)urea (1-(2-((4-fluorophenyl )ethynyl )—4'— (piper az in— 1—yl )-[l, 1' -biphenyl ] -4-yl )-3-(2-(pyridin- 3- yl)ethyl)urea) tert-butyl 4-(2'-((4-fluorophenyl)ethynyl)- 4'-(3-(2-(pyridin- 3- yl)ethyl)ureido)- [ 1 , 1' -biphenyl]- 4 -yl)piperazine- 1-carboxylate (tert- butyl 4-(2。1 -((4-f luorophenyl )ethynyl )-4 1 - (3- (2- (pyr idin- 3- y 1 )ethyl )ureido)-[l , 1 1 - bi pheny 1 ] -4-y 1 ) pi per az i ne-l-car boxy late, 0.047 g, 0.075 mmol , leq) was dissolved in DCM (0.5 ml), then trifluoroacetic acid (Tri fluoroacetic acid, 0.083 g, 0.75 mmol, 10 eq) was added and stirred at room temperature overnight. After completion of the reaction, the mixture was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in IfeO / Acetonitrile) to obtain the title compound (0.017 g, 43.6%). P NMR (400 MHz, DMSO) 5 8.76 (s, 1H) , 8.47 (d, J = 15.1 Hz, 2H), 7.81 (s, 1H), 7.69 (d, J = 7.6 Hz, 1H) , 7.52 (d, J = 8.2 Hz, 2H) , 7.48 - 7.43 (m, 2H), 7.35 (d, J = 7.6 Hz, 2H), 7.26 (dd, J = 16.4, 8.2 Hz, 3H), 7.04 (d, J = 8.1 Hz, 2H), 6.38 (s, 1H), 3.42 - 3.35 (m, 3H), 3.26 (s, 4H),

[0404] 3.07 (s, 4H), 2.81 (t, J = 6.7 Hz, 2H).; MS(ESI) m / z MH— 518.45 Example 11: 1-(3-((4-fluorophenyl)ethynyl)- 4-(2-methyl- 4-oxo- 3,4-dihydroquinazolin- 7-yl)phenyl)- 3-(2-(pyridin- 3 -yl)ethyl)urea (1-(3-((4-fluorophenyl)ethynyl)— 4— (2— methyl— 4— oxo— 3,4— dihydroquinazolin- 7-yl ) phenyl )-

[0405] Synthesis of 3-(2-(pyr idin-3-yl )ethyl )urea) (Synthetic scheme 6) Reaction Scheme 6. (a) PdC12(dppf), Bis(pinacolato)diboron, KOAc, Dioxane,

[0406] 60 °C, 16 hours (16h); (b) PdC12(dppf), K2CO3, 1,4-dioxane, water (H2O),

[0407] 90 °C, 4 h (4 h) Step 1 (a) Synthesis of 2-methyl- 7- (4, 4, 5, 5-tetramethyl- 1,3, 2-dioxaborolan- 2-yl)quinazolin- 4(3H)-one

[0408] 7-Bromo-2-methylquinazolin-4(3H)-one (0.24 g, 1 mmol, leq) was dissolved in Dioxane (10 ml, 0.1 M), and then bis(pinacolato)diboron (0.5 g, 2 mmol, 2 eq), potassium acetate (0.2 g, 2 mmol, 2 eq), and PdC12(dppf) (0.15 g, 0.2 mmol, 0.2 eq) were added and stirred at 60 °C for 16 h. The reaction mixture was concentrated under reduced pressure and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.075 g, 26%). Step 2 (b) Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(2-methyl- 4-oxo- 3,4-dihydroquinazolin- 7-yl)phenyl)- 3-(2-(pyridin- 3-yl)ethyl)urea) Compared to the synthesis of the compound of Example 9, in step 6) 2-methyl- 7-(4, 4, 5, 5-tetramethyl- 1,3, 2 The title compound (7.38 mg, 22%) was obtained in substantially the same manner as in step 6) of Example 9, except that 2-methyl-7- (4,4,5,5-tetramethyl- 1,3,2- di oxaborolan- 2-yl)quinazolin- 4(3H)-one was used. NMR (400 MHz, MeOD) 6 8.50 (s, 1H), 8.43 (d, J = 4.5 Hz, 1H), 8.26.

[0409] (d, J = 8.3 Hz, 1H), 7.90 (s, 1H), 7.85 - 7.76 (m, 3H), 7.48 - 7.41 (m, 3H), 7.37 (dd, J = 7.8, 5.7 Hz, 2H), 7.08 (t, J = 8.5 Hz, 2H) , 3.53 (t, J = 6.9

[0410] Hz, 2H), 2.94 (t, J= 7.0 Hz, 2H), 2.50 (s, 3H). MS(ESI) m / z MH+ 518 Example 12: Synthesis of 1-(2-(benzo[d]oxazol- 5-yl)ethyl)-3-(3-((4-fluorophenyl)ethynyl)-4-(pyridin- 4-yl)phenyl)urea (Scheme 7) Scheme 7. (a) Potassium (2-((tert-butoxycarbonyl)amino)ethyl)trifluoroborate, CS2CO3, Pd(0Ac)2, RuPhos, toluene / water; (b) TFA, DCM (c) TEA, THF, 24 h, reflux Step 1 (a) Synthesis of tert-butyl (2-(benzo[d]oxazol- 5-yl)ethyl)carbamate 5-bromobenzo[d]oxazole in toluene (6 mL) and water (2 mL) A mixture of oxazo 1 e, 0.2 g, 1 eq), potassium (2-((tert-butoxycarbonyl)amino)ethyl)trifluoroborate (0.28 g, 1.1 eq) and cesium carbonate (0.977 g, 3 eq) was degassed twice with nitrogen. Then, palladium (II) acetate (11 mg, 0.05 eq) and RuPhos (46 mg, 0.1 eq) were added, and the mixture was heated to 95 °C overnight under nitrogen. The reaction was cooled to room temperature, and water was added, and the mixture was extracted twice with EtOAc. The combined organic layers were then washed with brine, dried over anhydrous MgS04, filtered, and concentrated under reduced pressure. It was then purified (hexane / ethyl acetate = 1 / 5) to give the title compound (40 mg, yield Step 2 (b) Synthesis of 2-(benzo [ d ] oxazol-5 -yl)ethan-1-amine (2-( benzo [ d ] oxazo 1 -5 - yl )ethan-1-amine)

[0411] To a solution of tert-butyl (2-(benzo[d]oxazol-5-yl)ethyl)carbamate (0.04 g, 1 eq) in DCM (5 mL), TFA (0.11 mL, 10 eq) was added, and the mixture was stirred at room temperature for 2 h. The reaction mass was then concentrated under reduced pressure to afford crude 2-(benzo[d]oxazol-5-yl)ethan-1-amine, which was further dried under high vacuum. Step 3 (c) Synthesis of 1-(2-(benzo [d] oxazol-5-yl)ethyl)-3-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)urea (1-(2-(benzo [d] oxazo 1-5-yl)ethyl)-3-(3-((4-f luorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)urea) In comparison with the synthesis of the compound in Example 1, in Step 6) 2-(benzo [d] oxazol-5-yl)ethan-1-amine (2-) was used instead of 3-2-(aminoethyl)pyridine (3-(2-Aminoethyl)pyr idine).

[0412] The title compound was obtained in substantially the same manner as in Example 1, except that (benzo[d]oxazol-5-yl)ethan-1-amine) was used. In NMR (400 MHz, MeOD) δ 8.60 (d, J = 5.1 Hz, 2H), 8.45 (s, 1H), 7.77 - 7.61 (m, 5H), 7.45 - 7.34 (m, 5H), 7.10 (t, J = 8.6 Hz, 2H), 3.53 (t, J = 7.0 Hz, 2H), 3.01 (t, J = 7.0 Hz, 2H). Example 13: Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(piperazin-1-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-fluorophenyl)ethynyl)-4-(piperazin-1-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea) (Synthesis Scheme 8)

[0413] Scheme 8. (a) tert-Butyl piperazine-1-carboxylate, Cs2CO3, DMF, 80 °C, 24 h; (b) 1-Ethynyl-4-fluorobenzene, PdCl2(PPh3)2 fCui, TEA, ACN, 90 °C, 2 h; (c) Zn, NH4CI, 1,4 - dioxane, H2O, rt, 1 h; (d) Phenyl chloroformate, Pyridine, THF, 0 °C 2 h; (e) 2-(pyridin-3-yl)ethan-1-amine, TEA, THF, reflux, 4 h; (f) 4N HCl in dioxane, 1,4 - Dioxane, rt, 24 h Step 1 (a) Synthesis of tert-butyl 4-(2-iodo-4-nitrophenyl)piperazine-1-carboxylate

[0414] 1-Fluoro-2-iodo-4-nitrobenzene (0.1 g, 0.375 mmol, 1.0 eq) was dissolved in DMF (1 mL), and then CS2CO3 (0.15 g, 0.45 mmol,

[0415] 1.2 eq) and tert-butyl piperazine-1-carboxylate (0.07 g, 0.375 mmol, 1.0 eq) were added, and the mixture was stirred at 80 °C overnight. After completion of the reaction, it was extracted with EA and H2O. The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, EA / Hexane) (0.114 g, 70.4%) to obtain the title compound. 1H NMR (400 MHz, CDCl3) δ 8.71 (d, J = 2.2 Hz, 1H), 8.21 (dd, J = 8.8,

[0416] 2.2 Hz, 1H), 7.00 (d, J = 8.8 Hz, 1H), 3.70 - 3.61 (m, 4H), 3.11 - 3.03 (m, 4H), 1.49 (s, 9H). Step 2 (b) Synthesis of tert-butyl 4-(2-((4-fluorophenyl)ethynyl)-4-nitrophenyl)piperazine—l— carboxylate Tert-butyl 4-(2-iodo-4-nitrophenyl)piperazine—l— carboxylate (0.114 g, 0.263 mmol, 1 eq) was dissolved in ACN (3 ml, 0.1 M), followed by addition of TEA (0.08 ml, 0.579 mmol, 2.2 eq), PdC12(PPh3)2 (5.5 mg, 0.008 mmol, 3 mol%), Cui (2 mg, 0.011 mmol, 4 mol%) were added and stirred at room temperature for 5 minutes. 1-Ethynyl-4- f luorobenzene (0.045 ml, 0.395 mmol, 1.5 eq) was added and stirred at 90°C for 2 hours. After cooling to room temperature, the mixture was concentrated and purified by column chromatography (silica gel,

[0417] Purified by EA / HX) to obtain the title compound (0.109 g, 97.3%). 1H NMR (400 MHz, CDCl3) δ 8.39 (d, J = 2.5 Hz, 1H), 8.15 (dd, J = 9.1, 2.6 Hz, 1H), 7.52 (dd, J = 8.4, 5.4 Hz, 2H), 7.12 (t, J = 8.6 Hz, 2H), 6.95 (d, J = 9.1 Hz, 1H), 3.72 — 3.65 (m, 4H), 3.47 — 3.38 (m, 4H), 1.52 (s, 8H). Step 3 (c) Synthesis of tert-butyl 4-(4-amino-2-((4-fluorophenyl)ethynyl)phenyl)piperazine-1-carboxylate tert-Butyl 4-(2-((4-fluorophenyl)ethynyl)-4-nitrophenyl)piperazine-1-carboxylate (0.109 g, 0.256 mmol, 1 eq) was dissolved in 1,4-dioxane: water (H2O) (3:1 (v / v)) (3 ml, 0.4 M), then Zn (0.14 g, 2.56 mmol, 10 eq) and NH4Cl (0.137 g, 2.56 mmol, 10 eq) were added and stirred at room temperature for 1 hour. Washed with MeOH and filtered under reduced pressure. The filtrate was concentrated and then extracted with EA and water. The organic layer was dried over MgSO4, filtered, concentrated, and the next reaction in step 4) was carried out without purification. 1H NMR (400 MHz, CDCl3) δ 7.47 (dd, J = 8.2, 5.6 Hz, 2H), 7.26 — 7.26 (m, 1H), 7.05 (t, J = 8.5 Hz, 2H), 6.90 (s, 1H), 6.68 (d, J = 6.2 Hz, 1H), 3.64 (s, 4H), 3.09 (s, 4H), 1.48 (s, 9H). Step 4 (d) tert-Butyl 4-(2-((4-fluorophenyl)ethynyl)-4-.

[0418] ((phenoxycarbonyl)amino)phenyl)piperazine-1-carboxylate (tert-butyl 4~(2~((4~fluorophenyl)ethynyl)-4-((phenoxycarbonyl)amino)phenyl)piperazine—1—carboxylate) Synthesis tert-Butyl 4-(4-amino-2-((4-fluorophenyl)ethynyl)phenyl)piperazine-1-carboxylate (tert-butyl 4-(4-amino-2-((4-fluorophenyl)ethynyl)phenyl)piperazine—l—carboxylate, 0.114 g, 0.256 mmol, 1 eq) was dissolved in THF (3 ml, 0.1 M), and then at 0 °C, pyridine (0.04 ml, 0.512 mmol, 2 eq), phenyl chloroformate (0.049 ml, 0.384 mmol,

[0419] 1.5 eq) were added, and then it was stirred at room temperature for 2 hours. Water was added, stirred for 10 minutes, concentrated, and then purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.093 g, 70.5%). 1H NMR (400 MHz, CDCl3) δ 7.66 (s, 1H), 7.51 — 7.44 (m, 2H), 7.44 — 7.35 (m, 3H), 7.25 — 7.22 (m, 2H), 7.19 (d, J = 7.8 Hz, 3H), 7.06 (t, J =

[0420] 8.6 Hz, 2H), 6.87 (s, 1H), 3.67 (s, 4H), 3.18 (s, 4H), 1.48 (s, 9H). Step 5 (e) Synthesis of tert-butyl 4-(2-((4-fluorophenyl)ethynyl)- 4-(3-(2-(pyridin- yl)ethyl)ureido)phenyl)piperazine-1-carboxylate (tert-butyl 4-(2-((4-f luorophenyl )ethynyl )— 4— (3— (2— (pyridin— 3— yl )ethyl )ureido)phenyl )piperazine— 1-carboxylate tert-butyl 4-(2-((4-fluorophenyl)ethynyl)- 4-

[0421] ((phenoxycarbonyl)amino)phenyl)piperazine-1-carboxylate (tert- butyl 4- (2-((4- f luorophenyl )ethynyl ) -4 - ( ( phenoxy carbonyl 1 ) am i no ) pheny 1 )piperazine— 1— carboxylate, 0.093 g, 0.180 mmol , 1 eq) was dissolved in THF (2 ml , 0.1 M), and 2- (pyridin- 3 -yl)ethan- 1-amine (2- (pyridin- 3- yl)ethan- 1-amine, 0.042 ml , 0.36 mmol ,

[0422] 2 eq), TEA (0.075 ml, 0.54 mmol, 3 eq) were added and stirred at 80°C for 4 hours. The reaction mixture was concentrated and purified by column chromatography (silica gel, MC / MeOH) to obtain the title compound (0.089 g, 90.8%). P NMR (400 MHz, CDCls) 8 8.52 (s, 1H), 8.43 (d, J = 4.2 Hz, 1H),

[0423] 7.62 (d, J = 7.7 Hz, 1H) , 7.51 - 7.44 (m, 2H) , 7.41 (s, 1H) , 7.30 (d, J =

[0424] 6.9 Hz, 1H), 7.17 (d, J = 7.8 Hz, 1H) , 7.06 (t, J = 8.6 Hz, 2H) , 6.84 (d, J

[0425] = 8.7 Hz, 1H), 6.63 (s, 1H) , 5.02 (s, 1H) , 3.61 (s, 4H) , 3.56 — 3.47 (m, 2H) ,

[0426] 3.10 (s, 4H), 2.87 (t, J = 6.5 Hz, 2H), 1.48 (s, 9H). Step 6 (f) Synthesis of tert-butyl 4-(2-((4-fluorophenyl)ethynyl)-4-(piperazin-1-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-fluorophenyl)ethynyl)-4-(piperazin-1-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea) tert-butyl 4-(2-((4-fluorophenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)piperazine-1-carboxylate (tert-butyl 4-(2-((4-fluorophenyl)ethynyl)-4—(3—(2—(pyridin—3—yl) )ethyl )ureido )phenyl )piperazine— 1-carboxy late, 0.089g, 0.164mmol 1 , leq) was dissolved in 1,4-dioxane (1,4-dioxane, 2ml, 0.1M), then 4 N HCK4N HC1 in dioxane (0.16ml, 0.655mmol, 4eq) was added and stirred overnight. After washing with 1,4-dioxane and filtering under reduced pressure, the compound was obtained. P NMR (400 MHz, MeOD) 5 8.45 (s, 1H) , 8.39 (d, J = 4.4 Hz, 1H) , 7.77 (d, J = 7.8 Hz, 1H), 7.56 - 7.48 (m, 3H) , 7.39 (dd, J = 7.6, 5.0 Hz, 1H) , 7.24 (dd, J = 8.7, 2.4 Hz, 1H) , 7.14 (t, J = 8.7 Hz, 2H) , 6.95 (d, J = 8.8 Hz, 1H), 3.46 (t, J = 6.9 Hz, 2H) , 3.15 — 3.09 (m, 4H) , 3.05 — 2.99 (m, 4H) ,

[0427] 2.89 (t, J = 6.9 Hz, 2H). Example 14: Synthesis of 1-(4-(4-(3-aminobenzoyl)piperazin-1-yl)-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (Scheme 9) Scheme 9. (a) 3-((tert-butoxycarbonyl)amino)benzoic acid, DIPEA, HATU, DCM, room temperature (rt), 24 h (24 h); (b) 4N HC1 in dioxane, 1,4-dioxane, room temperature (rt), 24 h (24 h); Step 1 (a) tert-butyl (3-(4-(2-((4-fluorophenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)piperazine-1-carbonyl)phenyl)carbamate (tertbutyl (3-(4-(2-((4-fluorophenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)piperazine-1-carbonyl)phenyl)carbamate )ethynyl )- 4- (3- (2- (pyr idin- 3- yl )ethyl )ureido)phenyl )piperazine—l— carbonyl )phenyl )carbamate)

[0428] 3-((tert-butoxycarbonyl)amino)benzoic acid (3-((tert-butoxycarbonyl)amino)benzoic acid, 0.024g, 0.1mmol, leq) was dissolved in DCM (1ml, 0.3M) and then HATU (0.046g, 0.12mmol, 1.2eq) was added to DI PEA (0.05ml, 0.3mmol,

[0429] 3 eq), 1-(3-((4-fluorophenyl)ethynyl)-4-(piperazin-1-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-f luorophenyl)ethynyl)-4-(piperazin-1-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea, 0.044 g, 0.1 mmol, leq) was added and stirred at room temperature for 24 hours. The reaction mixture was concentrated and purified by column chromatography (silica gel, EA). (0.022g, 33.3%) P NMR (400 MHz, CDCls) 8 8.64 — 8.42 (m, 2H) , 8.02 (s, 1H) , 7.78 (d, J = 7.8 Hz, 1H), 7.72 (s, 2H) , 7.52 (s, J = 8.4 Hz, 1H) , 7.49 — 7.33 (m, 8H) , 7.20 (d, J = 7.6 Hz, 1H) , 7.11 - 7.03 (m, 4H) , 6.93 (s, 1H) , 6.85 (d, J = 8.4 Hz, 1H), 6.68 (d, J = 12.4 Hz, 2H) , 5.07 (s, 1H) , 3.98 (s, 2H) , 3.64 (s, 2H), 3.55 (s, 2H), 3.24 (s, 2H) , 3.13 (s, 2H) , 2.92 (s, 2H) , 1.54 (s, 9H) .Step 2 (b) Synthesis of tert-butyl (3-(4-(2-((4-fluorophenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)urea) tert-butyl (3-(4-(2-((4-fluorophenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)urea) tert-butyl (3-(4-.

[0430] (2-((4-f luorophenyl )ethynyl )- 4- (3- (2- (pyr idin- 3- yl )ethyl )ureido)phenyl )piperazine—l— carbonyl )pheny 1 ) car bamat e , 0.05g,

[0431] 0.075 mmol, leq) was dissolved in 1,4-dioxane (1,4-dioxane, 1 ml), then 4N HCK4N HC1 in dioxane (0.1 ml, 0.377 mmol, 5 eq) was added and stirred overnight. Saturated sodium bicarbonate (Sat. NaHCOs) aqueous solution was added, neutralized, and extracted with a colander. The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.018 g, 42.8%). P NMR (400 MHz, MeOD) 5 8.46 (s, 1H) , 8.41 (d, J = 4.2 Hz, 1H) , 7.79 (d, J = 7.8 Hz, 1H), 7.61 - 7.52 (m, 3H) , 7.44 - 7.39 (m, 1H) , 7.27 (dd, J = 8.8, 2.6 Hz, 1H), 7.22 - 7.12 (m, 3H), 7.01 - 6.97 (m, 1H), 6.80 (dd, J = 7.7, 1.9 Hz, 1H), 6.77 - 6.74 (m, 1H), 6.72 - 6.68 (m, 1H) , 3.95 (s, 2H) , 3.67 (s, 2H), 3.48 (t, J = 7.0 Hz, 2H) , 3.24 (s, 2H) , 3.13 (s, 2H) , 2.90 (t, J = 7.0 Hz, 2H). Example 15: Synthesis of 1-(4-(3-(2-aminoethyl)-2-methyl-4-oxo-3,4-dihydroquinazolin-7-yl)-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(4-(3-(2-aminoethyl)-2-methyl-4-oxo-3,4-dihydroquinazolin-7-yl)-3-((4-fluorophenyl)ethynyl)phenyl)— 3—(2—(pyridin— 3— yl)ethyl)urea) (Synthetic Reaction Scheme 10) Scheme 10. (a) tert-butyl (2-bromoethyl)carbamate, CS2CO3, DMF, 80 °C, 24 h (24 h); (b) 4N HC1 in dioxane, 1,4-dioxane, room temperature (rt), 24 h (24 h) Step 1 (a) tert-butyl (2- (7- (2- ((4-fluorophenyl)ethynyl)- 4- (3- (2-

[0432] (Pyridin-3-yl)ethyl)ureido)phenyl)-2-methyl-4-oxoquinazolin-3(4H)-yl)ethyl)carbamate (tert-butyl (2-(7-(2-((4-fluorophenyl)ethynyl)-4-(3-(2-

[0433] Synthesis of (pyr idin-3-yl )ethyl )ureido)phenyl )—2— methyl— 4— oxoquinazol in— 3(4H)— yl )ethyl )carbamate)

[0434] 1- (3-( (4-fluorophenyl)ethynyl)- 4- (2-methyl- 4-oxo- 3 , 4 -dihydroquinazoline-

[0435] 7 — (1)phenyl)one 3— (2— (pyridin— 3 —yl)ethyl)urea 0} (1-(3-( (4-f luorophenyl )ethynyl )-4- (2— methyl— 4— oxo— 3 ,4— dihydroquinazol in— 7— yl ) phenyl )— 3— (2— (pyridin— 3— yl ) ethyl )urea, 0.064 g, 0.124 mmol , 1.0 eq) was dissolved in DMF (1 mL), then CS2CO3 (0.081 g, 0.248 mmol , 1.2 eq) and tert-butyl (2-bromoethyl )carbamate (0.033 g, 0.15 mmol , 1.2 eq) were added and stirred at 80 °C overnight. After completion of the reaction, the mixture was extracted with EA and H20. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, DCM / MeOH) to obtain the title compound (0.049 g, 59.8%). Medium NMR (400 MHz, CDCI3) 5 8.49 (s, 2H), 8.24 (d, J = 8.3 Hz, 1H), 7.84 (s, 1H), 7.77 (d, J = 9.5 Hz, 1H), 7.68 (s, 2H), 7.54 - 7.48 (m, 1H), 7.37 (s, 2H), 7.35 — 7.29 (m, 3H), 7.04 (t, J = 8.7 Hz, 1H), 7.00 — 6.92 (m, 3H), 5.22 (s, 1H), 5.00 (t, J = 5.9 Hz, 1H), 4.27 (t, J = 6.0 Hz, 2H), 3.58

[0436] (s, 2H), 3.54 — 3.47 (m, 2H), 2.92 — 2.84 (m, 2H), 2.72 (s, 3H), 1.39 (s, 9H). Step 2 (b) Synthesis of 1-(4-(3-(2-aminoethyl)-2-methyl-4-oxo-3,4-dihydroquinazolin-7-yl)-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(4-(3-(2-aminoethyl)-2-methyl-4-oxo-3,4-dihydroquinazolin-7-yl)—3—((4-fluorophenyl)ethynyl)phenyl)—3—(2—(pyridin-3—yl)ethyl)urea) tert-butyl (2-(7-(2-((4-fluorophenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)urea) Tert-butyl (2- (7- (2- ((4- f luorophenyl )ethynyl )-4- (3- (2- (pyridin- 3- yl )ethyl )ureido)phenyl )— 2— methyl— 4— oxoquinazol in— 3(4H)— yl )ethyl)carbamate, 0.05 g, 0.075 mmol , leq) was dissolved in 1,4-dioxane (1,4-dioxane, 1 ml), and 4N HCK4N HC1 in dioxane, 0.1 ml, 0.377 mmol, 5 eq) was added and stirred overnight. After adding saturated sodium bicarbonate (Sat. NaHCOs) aqueous solution and neutralizing, extraction was performed with a kiln. The organic layer was dried over MgS04, filtered, concentrated, and purified by preparative HPLC (0.1% formic acid / Acetonitrile) to obtain the title compound (0.002 g, 0.5%). P NMR (400 MHz, MeOD) 5 8.50 (s, 5H), 8.44 (s, 1H), 8.30 (d, J = 8.3 Hz, 1H), 7.93 (s, 1H), 7.80 (dd, J = 12.0, 5.0 Hz, 3H) , 7.51 — 7.36 (m, 5H), 7.08 (t, J = 8.7 Hz, 2H), 4.49 (t, J = 6.2 Hz, 2H) , 3.62 (s, 1H) , 3.53 (t, J = 7.0 Hz, 2H), 3.41 — 3.36 (m, 2H) , 2.94 (t, J = 6.9 Hz, 2H) , 2.73 (s, 3H) . Example 16: N-(2-(3-aminopiperidin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (N-(2-(3-aminopiperidin— 1— yl)ethyl)— 4— ((5— (3— (2— (pyridin— 3— yl)ethyl)ureido)— 2 —.

[0437] Synthesis of (pyridin-4-yl)phenyl)ethynyl)benzamide (Synthetic scheme 11) Scheme 11. Reagents and conditions: (a) HATU, DIPEA,

[0438] DMF, room temperature (rt), overnight; (b) Phenyl chloroformate, pyridine, THF, 0°C, 2 h; (c) 2-(pyridin-3-yl)ethan-1-amine, TEA, THF, reflux, 24 h; (d) PdC12(PPh3)2, Cui, TEA, ACN, 60°C, 5 h; (e) 4N HC1 in dioxane, 1,4-dioxane, room temperature (rt), 2 h Step 1 (a) tert-butyl Synthesis of (1-(2-(4-ethynylbenzamido)ethyl)piperidin-3-yl)carbamate (tert-butyl (1-(2-(4-ethynylbenzamido)ethyl)piperidin-3-yl)carbamate)

[0439] 4-Ethynylbenzoic acid (0.15 g, 1 mmol, 1 eq) was dissolved in DMF (3 ml, 0.3 M), and tert-butyl (1-(2-aminoethyl)piperidin-3-yl) carbamate (0.243 g, 1 mmol, 1.5 eq), HATU (0.456 g, 1.2 mmol, 1.2 eq), and DI PEA (0.52 ml, 3 mmol, 3 eq) were added. The mixture was stirred at room temperature overnight. The mixture was washed with water and filtered to obtain the title compound (0.371 g, 74%). Medium NMR (400 MHz, CDC13) 8 7.74 (d, J = 8.3 Hz, 2H), 7.55 (d, J = 8.3 Hz, 2H), 6.76 (s, 1H), 4.72 (s, 1H), 3.72 (s, 1H), 3.53 (dt, J = 10.5, 5.3 Hz, 2H), 2.75 (s, 1H), 2.56 (t, J = 5.3 Hz, 2H), 2.36 - 2.09 (m, 2H), 1.84 — 1.69 (m, 3H), 1.43 (s, 9H). Step 2 (b) Synthesis of phenyl (3-iodo-4-(pyridin-4-yl)phenyl)carbamate (phenyl (3-iodo-4-(pyridin-4-yl)phenyl)carbamate)

[0440] 3-Iodo-4-(pyridin-4-yl)aniline (0.592 g, 2 mmol, 1 eq) was dissolved in THF (20 ml, 0.1 M). Then, pyridine (0.32 ml, 4 mmol, 2 eq) and phenyl chloroformate (0.38 ml, 3 mmol, 1.5 eq) were added dropwise at 0 °C while stirring. After completion of the reaction, it was concentrated and extracted with EA and water. The organic layer was dried over MgSO₄ and then the next reaction in step 3 was carried out without purification. ¹H NMR (400 MHz, CDCl₃) δ 8.79 (d, J = 6.0 Hz, 2H), 8.20 (s, 1H), 7.91 (d, J = 6.2 Hz, 2H), 7.63 (d, J = 8.0 Hz, 1H), 7.38 - 7.33 (m, 3H), 7.24 (d, J = 8.6 Hz, 1H), 7.13 (d, J = 7.6 Hz, 2H). Step 3 (c) Synthesis of 1-(3-iodo-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea Phenyl (3-iodo-4-(pyridin-4-yl)phenyl)carbamate (1.14 g, 2 mmol, 1 eq) was dissolved in THF (20 ml, 0.1 M). Then, 2-(pyridin-3-yl)ethan-1-amine (0.47 ml, 4 mmol, 2 eq) and TEA (0.84 ml, 6 mmol, 3 eq) were added and stirred at 80 °C for 4 hours.The reaction mixture was concentrated and purified by column chromatography (silica gel, MC / MeOH) to afford the title compound (0.552, 62%). 1H NMR (400 MHz, CDCl3) δ 8.62 (d, J = 5.9 Hz, 2H), 8.40 (s, 2H),.

[0441] 7.93 (d, J = 2.0 Hz, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.51 (s, 1H), 7.41 (dd,

[0442] J = 8.4, 2.1 Hz, 1H), 7.31 - 7.27 (m, 1H), 7.24 (s, 1H), 7.12 (d, J = 8.3

[0443] Hz, 1H), 5.42 (t, J = 5.6 Hz, 1H), 3.58 (q, J = 6.2 Hz, 2H), 2.89 (t, J =

[0444] 6.5 Hz, 2H). Step 4 (d) tert-Butyl (1-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-

[0445] 2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperidin-3-yl)carbamate (tert-butyl (1-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperidin-3-yl)carbamate) synthesis

[0446] 1-(3-iodo-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea(!-(3— iodo— 4— (pyridin— 4— yl)phenyl)— 3— (2— (pyridin— 3— yl)ethyl)urea, 0.044 g, 0.1 mmol, 1 eq) was dissolved in ACN (1 ml, 0.1 M), then TEA (0.03 ml, 0.22 mmol, 2.2 eq), PdC12(PPhs)2 (2.1 mg, 0.003 mmol, 3 mol%), and Cui (0.8 mg, 0.004 mmol, 4 mol%) were added and stirred at room temperature for 5 min. tert-butyl (1-(2-(4- After adding tert-butyl (l-(2-(4-ethynylbenzamido)ethyl)piperidin— 3— yl)carbamate (0.045 g, 0.12 mmol, 1.2 eq), the mixture was stirred at 90°C for 2 hours. After cooling to room temperature, the mixture was concentrated and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.109 g, 97.3%). P NMR (400 MHz, CDC13) 8 8.57 (s, 4H), 8.00 (s, 1H), 7.67 (d, J =

[0447] 8.1 Hz, 2H), 7.62 (t, J = 8.2 Hz, 2H) , 7.45 (s, 3H) , 7.31 (s, 1H) , 7.25 -

[0448] 7.19 (m, 4H), 5.94 (s, 1H) , 5.05 (s, 1H) , 3.80 (s, 1H) , 3.68 - 3.50 (m, 5H) ,

[0449] 3.03 (s, 1H), 2.91 (t, J = 6.5 Hz, 2H), 2.82 (s, 3H), 2.51 (s, 2H), 2.02 (s,

[0450] 10H), 1.83 (s, 3H), 1.72 (s, 2H), 1.42 (d, J = 19.1 Hz, 12H), 1.25 (s, 2H). Step 5 (e) Synthesis of N-(2-(3-aminopiper idin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (tert-butyl (1-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide) tert-butyl (1-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide) -yl)carbamate (tert- butyl (1-(2- (4-((5-(3-(2-(pyr idin-3-yl)ethyl)ureido)-2-(pyr idin-4- yl)phenyl)ethynyl)benzamido)ethyl)piperidin— 3— yl)carbamate, 0.03g,

[0451] 0.044 mmol 1 , leq) was dissolved in 1,4-dioxane (1,4-dioxane, 1 ml), then 4N HC1 in dioxane (4N HC1 in dioxane, 0.1 ml, 0.436 mmol , lOeq) was added and stirred at room temperature for 2 hours. The title compound (0.014 g, 50%) was obtained by washing with 1,4-dioxane and filtering under reduced pressure. NMR (400 MHz, MeOD) 6 8.89 (s, 2H), 8.86 (s, 1H), 8.75 (s, 1H),

[0452] 8.60 (d, J = 8.1 Hz, 1H), 8.41 (d, J = 5.3 Hz, 2H), 8.08 - 8.02 (m, 1H),

[0453] 7.97 (s, 1H), 7.96 - 7.94 (m, 2H), 7.68 - 7.64 (m, 1H), 7.63 - 7.59 (m, 1H), 7.59 - 7.55 (m, 2H), 3.90 (s, 1H), 3.85 (t, J = 5.6 Hz, 2H), 3.73 (s, 2H),

[0454] 3.61 (t, J = 6.8 Hz, 2H), 3.49 (t, J = 5.7 Hz, 2H), 3.14 (t, J = 6.7 Hz, 3H),

[0455] 2.26 - 2.13 (m, 2H), 2.09 - 1.96 (m, 1H), 1.80 - 1.66 (m, 1H), 1.44 (s, 1H). Example 17: Synthesis of 1-(4-(3-(2-aminoethyl)-2-methyl-4-oxo-3,4-dihydroquinazolin-7-yl)-3-((4-fluorophenyl)ethynyl)phenyl)-3-(imidazole [1,2-a]pyridin-7-ylmethyl)urea) (Synthetic Scheme 12)

[0456] Scheme 12. (a) Pd(dppf)C12, K2CO3, 1,4-dioxane, water (H2O), 90 °C overnight; (b) tert-butyl (2-bromoethyl)carbamate, CS2CO3, DMF, 80 °C for 24 h; (c) 4 N HC1 in dioxane, 1,4-dioxane, room temperature (rt), 24 h Step 1 (a) 1- (3- ( (4-fluorophenyl)ethynyl )-4- (2-methyl- 4-oxo- 3 , 4-dihydroquinazolin- 7 -yl)phenyl)- Synthesis of 3-(Imidazo [1,2- a]pyridin- 7 -ylmethyl)urea (1-(3- ((4— fluorophenyl )ethynyl )—4— (2— methyl— 4— oxo— 3,4— dihydroquinazol in— 7 — yl ) phenyl )-3-( imidazo[l,2- a]pyr idin- 7- ylmethyl )urea) In comparison with the synthesis of the compound of Example 11, in step 2) 1-(3- ((4-fluorophenyl)ethynyl)-4 -iodophenyl)- 3- (2- (pyridin- 3 -yl)ethyl)urea) was used instead of 1- The title compound (0.034 g, 31.2%) was prepared in substantially the same manner as in step 6 of Example 11, except that (3-((4-fluorophenyl)ethynyl)-4-iodophenyl)-3-(imidazo[l,2-a]pyridin-7-ylmethyl)urea) was used.

[0457] 1H NMR (400 MHz, MeOD) 5 8.43 (d, J = 7.1 Hz, 1H) , 8.26 (d, J = 8.3

[0458] Hz, 1H), 7.91 (s, 1H), 7.85 (d, J = 2.0 Hz, 1H) , 7.82 (s, 1H) , 7.78 (d, J =

[0459] 8.2 Hz, 1H), 7.57 - 7.50 (m, 3H) , 7.49 - 7.45 (m, 1H) , 7.36 (dd, J = 8.6,

[0460] 5.5 Hz, 2H), 7.07 (t, J = 8.8 Hz, 2H) , 6.97 (d, J = 7.1 Hz, 1H) , 4.52 (s,

[0461] 2H), 2.50 (s, J = 5.9 Hz, 3H) . Step 2 (b) Synthesis of tert-butyl (2-(7-(2-((4-fluorophenyl)ethynyl)-4-(3-(imidazo[l,2-a]pyridin-7-ylmethyl)ureido)phenyl)-2-methyl-4-oxoquinazolin-3(4H)-yl)ethyl)carbamate (tert-butyl (2-(7-(2-((4-f luorophenyl)ethynyl)-4-(3-(imidazo[l,2— a]pyridin-7-ylmethyl)ureido)pheny 1 )-2-methyl-4-oxoquinazolin-3(4H)-yl)ethyl)carbamate) Comparing with the synthesis of the compound of Example 15, 1-(3-((4-fluorophenyl)ethynyl)-4-(2 -methyl- 4-oxo- 3 , 4 -dihydroquinazolin- 7 -yl)phenyl)- 3-(2- (pyridin- 3 -yl)ethyl)urea (1- (3- ((4- f luorophenyl )ethynyl )-4- (2- methyl- 4-oxo- 3 , 4-dihydroquinazolin- 7-yl )phenyl )—3— (2— (pyridin— 3— yl )ethyl )urea) instead of 1- (3-((4-fluorophenyl)ethynyl)- 4- (2-methyl- 4 -oxo- 3 , 4 -dihydroquinazolin- 7-yl)phenyl)- 3- (imidazo [1,2- a]pyridin- 7 -ylmethyl)urea (1- (3-((4- f luorophenyl )ethynyl )—4— (2— methyl— 4— oxo— 3 , 4— dihydroquinazol in— 7— yl ) phenyl )-

[0462] The title compound (23 mg, 56.1%) was obtained in substantially the same manner as in the synthesis in step 1) of Example 15, except that 3-( imidazo[l , 2-a]pyr idin-7-ylmethyl )urea) was used. Step 3 (c) Synthesis of 1-(4-(3-(2-aminoethyl)-2-methyl-4-oxo-3,4-dihydroquinazolin-7-yl)-3-((4-fluorophenyl)ethynyl)phenyl)-3-(imidazole [1,2-a]pyridin-7-ylmethyl)urea (1-(4-(3-(2-aminoethyl)-2-methyl-4-oxo-3,4-di hydr oquinazol in-7-y 1)-3-((4-fluorophenyl)ethynyl 1)pheny 1)-3-(imidazole [1,2-a]pyridin-7-ylmethyl )urea) Compared to the compound synthesis of Example 15 in Step 2), tert-butyl (2-(7-(2-( (4-fluorophenyl)ethynyl)- 4- (3- (2- (pyridin- 3 -yl)ethyl)ureido)phenyl)- 2-methyl- 4-oxoquinazolin- 3(4H)-yl)ethyl)carbamate (tert- butyl (2- (7- (2-( (4- f luorophenyl )ethynyl )— 4— (3— (2— (pyridin— 3— yl )ethyl )ureido)phenyl )—2—methyl—

[0463] The same procedure as in step 2) of Example 15 except that tert-butyl (2- (7- (2-( (4-fluorophenyl)ethynyl)-4- (3- (imidazo[1,2-a]pyridin-7-ylmethyl)ureido)phenyl)-2-methyl-4-oxoquinazolin-3(4H)-yl)ethyl)carbamate) was used instead of 4-oxoquinazol in-3(4H)-yl)ethyl)carbamate) The title compound (10 mg, 86.3%) was obtained in substantially the same manner as the synthesis.

[0464] 1H NMR (400 MHz, MeOD) 5 8.79 (d, J = 7.0 Hz, 1H) , 8.43 (d, J = 8.3

[0465] Hz , 1H) , 8.21 (s , 1H) , 8.07 - 8.00 (m, 2H) , 7.98 (s , 1H) , 7.92 (d, J = 1.9 Hz, 1H), 7.88 (s, 1H), 7.62 - 7.57 (m, 1H), 7.57 - 7.47 (m, 2H), 7.45 - 7.35 (m, 2H), 7.11 (t, J = 8.7 Hz, 2H), 4.67 (s, 2H), 4.60 (t, J = 6.1 Hz, 2H), 3.48 (t, J = 6.0 Hz, 2H), 2.99 (s, 3H). Example 18: Synthesis of 1-(3-(1H-pyrazol-4-yl)propyl)-3-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)urea (1-(3-(1H-pyrazol-4-yl)propyl)—3—(3—((4—f luorophenyl)ethynyl)—4—(pyridin—4—yl)phenyl)urea) (Synthetic Scheme 13) Scheme 13. (a) 3-(1H-pyrazol-4-yl)propan-1-amine, TEA, THF, reflux, 3 h; (b) 1-ethynyl-4-fluorobenzene, PdCl2(PPh3)2 f Cui, TEA, ACN, 90 °C, 2 hrs Step 1 (a) Synthesis of 1-(3-(1H-pyrazol-4-yl)propyl)-3-(3-iodo-4-(pyridin-4-yl)phenyl)urea (1-(3-(1H-pyrazol-4-yl)propyl)-3-(3-iodo-4-(pyridin-4-yl)phenyl)urea) Compared with the synthesis of the compound of Example 16, in Step 3) 2-(pyridin-3-yl)ethane-

[0466] It was prepared in substantially the same manner as Example 16, except that 3-(1H-pyrazol-4-yl)propan-1-amine was used instead of 1-amine (2-(pyr idin-3-yl)ethan-1-amine). (0.1 g, 93.5%) P NMR (400 MHz, MeOD) 5 8.60 (d, J = 5.5 Hz, 2H), 8.21 (d, J = 2.1 Hz, 1H), 7.53 - 7.42 (m, 5H), 7.24 (d, J = 8.4 Hz, 1H) , 3.37 (s , 1H) , 3.28 - 3.26 (m, 2H) , 2.61 (t , J = 7.6 Hz , 2H) , 1.90 - 1.78 (m, 2H) . Step 2 (b) Synthesis of 1-(3-(1H-pyrazol-4-yl)propyl)-3-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)urea

[0467] 1-(3-(1H-pyrazol-4-yl)propyl)-3-(3-iodo-4-(pyridin-4-yl)phenyl)urea (1-(3-(IH-pyr azol -4-yl) pr opy 1)-3-( 3- iodo-4-( pyr idi n- 4- yl )phenyl )urea, 0.1 g, 0.22 mmol, 1 eq) was dissolved in ACN (2 ml, 0.1 M), and then TEA (0.07 ml, 0.48 mmol, 2.2 eq), PdC12(PPhs)2 (4.6 mg, 0.007 mmol, 3 mol%), and Cui (2 mg, 0.009 mmol, 4 mol%) were added and stirred at room temperature for 5 min. 1-Ethynyl-4-fluorobenzene (1—ethynyl— 4— f luorobenzene , 0.030 ml , 0.27 mmol , 1.2 eq) was added and stirred at 90°C for 2 hours. After cooling to room temperature, the mixture was concentrated and purified by column chromatography (C18 , 0.1% formic acid in H2O / acetonitrile) to obtain the title compound (2.8 mg, 3%). P NMR (400 MHz, MeOD) 5 8.62 (s, 2H), 7.82 (d, J = 2.2 Hz, 1H), 7.74 (d, J = 6.0 Hz, 2H), 7.70 - 7.63 (m, 2H), 7.61 - 7.57 (m, 1H) , 7.53 - 7.49 (m, 1H), 7.47 - 7.44 (m, 1H), 7.42 - 7.38 (m, 2H), 7.12 (t, J = 8.8 Hz, 2H), 3.28 (t, J = 7.0 Hz, 2H), 2.62 (t, J = 7.5 Hz, 2H), 1.90 - 1.81 (m, 2H).Example 19: Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(isoquinolin-6-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-fluorophenyl)ethynyl)-4-(isoquinolin-6-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea) (Synthesis Scheme 14). Scheme 14. (a) NaNO2, KI, pTSA, ACN, 0 °C, 1 hour (1h); (b) Pd(dppf)Cl2, K2CO3, 1,4-dioxane, water (H2O), 90 °C 3 hours (3h); (c) Fe, NH4Cl, EtOH, water (H2O), reflux, 1 hour (1h); (d) phenyl chloroformate, pyridine, THF, 0 °C to room temperature (rt), 2 hours (2h); (e) 2-(pyridin-3-yl)ethan-1-amine, TEA, THF, room temperature (rt), reflux step 1 (a) Synthesis of 2-((4-fluorophenyl)ethynyl)-1-iodo-4-nitrobenzene

[0468] 2-((4-fluorophenyl)ethynyl)-4-nitroaniline (2-((4-f 1 uor opheny 1 ) et hyny 1 ) -4-nitr oan i 1 i ne , 0.1 g, 0.39 mmol , 1 eq) was dissolved in ACN (4 ml , 0.1 M), pTSA (0.2 g, 1.17 nmol, 3 eq) was added, and then NaN02 (0.03 g, 0.408 mmol, 1.02 eq) dissolved in 0.15 ml of water and KI (0.13, 0.78 mmol, 2 eq) dissolved in 0.15 ml of water were added at 0 °C. After reacting at room temperature for 1 hour, saturated sodium bicarbonate (Sat. NaHCOs) aqueous solution was added, neutralized to pH 8 or higher, and extracted with a distilled water. The organic layer was dried over MgS04, filtered, and concentrated to obtain the target compound as a crude product. Blood NMR (400 MHz, CDC13) 8 8.31 (d, J = 2.6 Hz, 1H), 8.07 (d, J = 8.7 Hz, 1H), 7.83 (dd, J = 8.7, 2.7 Hz, 1H), 7.64 - 7.59 (m, 2H), 7.10 (t, J = 8.6 Hz, 2H) Step 2 (b) Synthesis of 6-(2-((4-fluorophenyl)ethynyl)-4-nitrophenyl)isoquinoline (6-(2-((4-fluorophenyl)ethynyl)-4-nitrophenyl)isoquinoline)

[0469] 2-((4-fluorophenyl)ethynyl)-1-iodo-4-nitrobenzene (2-((4-fluorophenyl)ethynyl)—l— iodo— 4— nitrobenzene, 0.14 g, 0.38 mmol, 1 eq) was dissolved in 1,4-dioxane: water (H20) (3:l(v / v), 2 ml, 0.2 MH], and then isoquinol in-6-ylboronic acid (0.066 g, 0.38 mmol, 1 eq), Pd(dppf)C12 (0.056 g, 0.076 mmol, 0.2 eq), K2CO3 (0.116 g, 0.836 mmol, 2.2 After adding eq), degassing was performed with nitrogen gas (due gas). The reaction solution was heated to 90 °C and stirred for 3 hours. After completion of the reaction, it was filtered through a celite filter and extracted with EA and H2O. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.032 g, 23%). P NMR (400 MHz, MeOD) 5 9.37 (s, 1H), 8.58 — 8.52 (m, 2H), 8.38 (d, J = 8.5 Hz, 1H), 8.31 (s, 2H), 8.06 (d, J = 8.5 Hz, 1H), 7.97 (d, J = 5.7 Hz, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.38 — 7.30 (m, 2H), 7.08 (t, J = 7.9 Hz, 2H). MS(ESI) m / z MH+ 369 Step 3 (c) Synthesis of 3-((4-fluorophenyl)ethynyl)-4-(isoquinolin-6-yl)aniline (3-((4-f luorophenyl )ethynyl )—4—( isoquinolin— 6— yl )ani 1 ine)

[0470] A mixture of 6-(2-((4-fluorophenyl)ethynyl)-4-nitrophenyl)isoquinoline (6-(2-((4- f 1 uor opheny 1 ) et hyny 1 ) -4-nitr opheny 1 ) i soqu i no 1 i ne , 0.032 g, 0.087 mmol 1 , 1.0 eq), Fe (0.024 g, 0.434 mmol, 5 eq) and NH4C1 (0.023 g, 0.434 mmol, 5 eq) was stirred at 80°C for 1 h. The catalyst was removed by filtration through Celite, and the filtrate was concentrated under reduced pressure. Water was added, and extraction was performed with a sieve, and the organic layer was concentrated under reduced pressure. The target compound was obtained as a crude product. Step 4 (d) Synthesis of phenyl (3-((4-fluorophenyl)ethynyl)-4-(isoquinolin-6-yl)phenyl)carbamate) Compared to the synthesis of the compound in Example 1, in Step 5) 3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)aniline (3-((4-f luorophenyl)ethynyl)-4-(isoquinolin-6-yl)aniline) was used instead of 3-((4-f luorophenyl)ethynyl)-4-(isoquinolin-6-yl)aniline )ani 1 ine) was used, the title compound was prepared in substantially the same manner as the synthesis in step 5) of Example 1.

[0471] MS(ESI) m / z MH+ 459 Step 5 (e) Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(isoquinolin-6-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-f luorophenyl)ethynyl)-4-(isoquinolin-6-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea) Compared to the synthesis of the compound in Example 1, in Step 6) phenyl (3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)carbamate was used instead of phenyl (3-((4-fluorophenyl)ethynyl)-4-(isoquinolin-6-yl)phenyl)carbamate (phenyl The title compound was prepared in substantially the same manner as in step 6) of Example 1, except that (3-( (4- f luorophenyl )ethynyl )—4—( isoquinol in— 6— yl )phenyl ) carbamate) was used. P NMR (400 MHz, MeOD) 5 9.29 (s, 1H), 8.53 — 8.46 (m, 2H), 8.43 (s, 1H), 8.23 ​​— 8.16 (m, 2H), 8.02 (d, J = 8.4 Hz, 1H), 7.90 (d, J = 5.3 Hz, 1H), 7.86 - 7.79 (m, 2H), 7.54 - 7.47 (m, 2H), 7.47 - 7.40 (m, 1H), 7.34 - 7.26 (m, 2H), 7.05 (t, J = 8.2 Hz, 2H), 3.54 (t, J = 6.8 Hz, 2H) , 2.95 (t, J = 6.4 Hz, 2H) . MS(ESI) m / z MH+ 487 Example 20: 1- (3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(6-nitropyridin-3-yl)ethyl)urea (1- (3-((4- f luorophenyl)ethynyl)-4-

[0472] Synthesis of (pyr idin-4-yl ) phenyl )-3- (2- (6- nitropyr idin-3- yl ) ethyl ) urea) (Synthetic Scheme 15) Scheme 15. Reagents and conditions: (a) Pd(dppf)C12, CS2CO3, toluene, water (H2O), 80 °C for 16 h; (b) 4 N HC1 in dioxane, 1,4-dioxane, room temperature (rt), for 16 h; (c) triethylamine (TEA), THF, room temperature (rt), overnight. Step 1 (a) Synthesis of tert-butyl (2-(6-nitropyr idin- 3-yl)ethyl)carbamate

[0473] 5-Bromo-2-nitro-pyridine (1 g, 4.93 mmol, 1.0 eq) was dissolved in toluene (20 ml, 0.25 M) and water (H2O, 5 ml, IM), and then potassium 2-(Boc-aminoethyl)trifluoroborate (1.36 g, 5.41 mmol, 1.1 eq), Pd(dppf)C12 (0.302 g, 0.369 mmol, 0.07 eq), and CS2CO3 (4.82 g, 14.8 mmol, 3 eq) were added. The reaction mixture was heated to 80°C and stirred for 16 h. After the reaction was completed, it was cooled to room temperature and extracted with EA, 0. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, EA / HX) to obtain the title compound (0.842 g, 64%). Step 2 (b) Synthesis of 2-(6-nitropyridin-3-yl)ethan-1-amine Tert-butyl (2-(6-nitropyridin-3-yl)ethyl)carbamate (0.84 g, 3.15 mmol, 1 eq) was dissolved in 1,4-dioxane (1,4-di oxane, 6 ml, 0.5 MH), and 4N HC1 (4N HC1 in dioxane, 8 ml, 31.5 mmol, 10 eq) was added, followed by stirring at room temperature for 16 hours. After completion of the reaction, the mixture was concentrated, and diethyl ether was added. The resulting The crystals were washed with ether and filtered under reduced pressure to obtain the title compound (0.507 g, 79%).Step 3 (c) Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(6-nitropyridin-3-yl)ethyl)urea ( 1-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(6-nitropyridin-3-yl)ethyl)urea) phenyl (3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)carbamate ( phenyl ( 3-((4-f 1 uoropheny 1 )ethynyl 1 )-4-( pyr idin-4- y 1 )pheny 1 ) carbamate , 0.081 g, 0.20 mmol, 1 eq) was dissolved in THF (1 ml, 0.2 M), then 2-(6-nitropyridin-3-yl)ethan-1-amine (2-(6-ni tropyr idin-3- y 1)ethan-l-amine, 0.082 mg, 0.4 mmol, 2 eq) and TEA (0.84 ml, 0.6 mmol, 3 eq) were added and stirred at room temperature for 16 h. After completion of the reaction, the mixture was concentrated and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.113 g, 23%). P NMR (400 MHz, CDCls) 8 8.67 (d, J = 4.2 Hz, 2H), 8.49 (s, 1H), .

[0474] 8.17 (d, J = 8.2 Hz, 1H), 7.93 (d, J = 8.2 Hz, 1H) , 7.74 (s, 1H) , 7.64 (d, J = 5.1 Hz, 3H), 7.39 (d, J = 8.5 Hz, 1H) , 7.31 (d, J = 8.2 Hz, 2H) , 6.99 (t, J = 8.2 Hz, 2H), 5.77 (s, 1H) , 3.65 (d, J = 6.1 Hz, 2H) , 3.08 (t, J = 6.3 Hz, 2H); MS(ESI) MH +482.4 Example 21: Synthesis of 1-(2-(6-aminopyridin-3-yl)ethyl)-3-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)urea (1-(2-(6-aminopyr idin-3-yl)ethyl)—3—(3—((4—f luorophenyl)ethynyl)—4—(pyr idin—4—yl)phenyl)urea) (Synthetic Reaction Scheme 16) Scheme 16. Reagents and conditions: (a) Zn, NH4CI,

[0475] 1,4-dioxane, water (H2O), room temperature (rt), overnight

[0476] 1-(3-((4-fluorophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(6-nitropyridin-3-yl)ethyl)urea (1-(3-((4-f luorophenyl )ethynyl )-4-(pyr idin-4- yl)phenyl)— 3— (2— (6— nitropyridin— 3— yl)ethyl)urea, 0.048 g, 0.1 mmol , 1 eq)

[0477] 1.4-Dioxane: Dissolve in water (H2O)(3:l(v / v)) (0.5 ml, 0.2 M), then add Zn (0.065 g, 1 mmol, 10 eq) and NH4CI (0.053 g, 1 mmol, 10 eq) and stir at room temperature overnight. Wash with MeOH, filter under reduced pressure, concentrate, and purify by reverse phase column chromatography (0.1% formic acid in H2O / ACN) to obtain the title compound (0.001 g, 2.4%). P NMR (400 MHz, MeOD) 5 8.62 (s, 2H) , 7.80 (s, 2H) , 7.73 (d, J = 4.5 Hz, 2H), 7.61 (d, J = 8.7 Hz, 1H) , 7.46 (dd, J = 18.2, 8.6 Hz, 2H) , 7.42 - 7.37 (m, 2H), 7.12 (t, J = 8.3 Hz, 2H), 6.74 (d, J = 8.5 Hz, 1H), 3.45 (t, J = 6.8 Hz, 2H), 2.75 (t, J = 6.8 Hz, 2H); MS MH' 450.39 Example 22: N- Synthesis of (2-(piperazin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (N-(2-(piperazin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide) (Synthetic Scheme 17)

[0478] Scheme 17. Reagents and conditions: (a) tert-butyl 4-(2-aminoethyl)piperazine—l— carboxylate, HATU, DIPEA, DMF, room temperature (rt), overnight; (b) NaN02, KI, HC1, H20, 5 °C, 1 h; (c) PdC12(PPh3)2, Cui, TEA, ACN, 60 °C, 5 h; (d) Zn, NH4Cl, 1,4-dioxane, water (H2O), room temperature (rt), overnight; (e) phenyl chloroformate , pyridine, THF, 0 °C to room temperature (rt), 2 hrs; (f) 2-(pyridin-3-yl)ethan-l-amine, TEA, THF, room temperature (rt), overnight; (g) TFA, DCM, room temperature (rt), overnight Step 1 (a) Synthesis of tert-butyl 4-(2-(4-ethynylbenzamido)ethyl)piperazine-1-carboxylate

[0479] 4-Ethynylbenzoic acid (0.25 g, 1.71 mmol, 1 eq) was dissolved in DMF (11 ml, 0.3 M), and tert-butyl 4-(2-aminoethyl)piperazine-1-carboxylate (0.589 g, 2.57 eq, 1.5 eq), HATU (0.977 g, 2.57 mmol, 1.5 eq), and DI PEA (0.663 g, 5.13 mmol, 3 eq) were added and stirred at room temperature overnight. The organic layer was extracted with doyu and Seong0, dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.382 g, 63%). Step 2 (b) Synthesis of 4-(2-iodo-4-nitrophenyl)pyridine

[0480] 5-Nitro-2-(pyridin-4-yl)aniline (5-nitro-2-(pyridin-4-yl)aniline, 1.00 g, 4.64 mmol, 1 eq) was added to 4.5 ml of water and 4.5 ml of cone. HC1 was added, and then NaN02 (0.326 g, 4.73 mmol, 1.0 eq) dissolved in 4.5 ml of water was slowly added dropwise while stirring at 5 °C. After the addition, the mixture was stirred for 10 minutes, and KI (1.464 g, 8.82 mmol, 2 eq) dissolved in 4.5 ml of water was added. The mixture was warmed to room temperature and stirred for 30 minutes. After adding aqueous K2CO3 solution, the mixture was neutralized to pH 8 or higher, and extracted with DCM. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, Hex / EA) to obtain the title compound (0.952 g, 62.9%). Step 3 (c) tert-butyl 4- (2- (4- ( (5 -nitro- 2- (pyridin- 4-yl) phenyl) ethynyl) benzamido) ethyl) piperazine-1-carboxylate (tert- butyl 4- (2- (4-

[0481] Synthesis of ((5— nitro— 2— (pyr idin— 4— yl ) phenyl )ethynyl )benzamido)ethyl )piperazine— 1— carboxylate)

[0482] 4-(2-iodo-4-nitrophenyl)pyridine (0.232 g, 0.71 mmol, 1 eq), tert-butyl 4-(2-(4-ethynylbenzamido)ethyl)piperazine-1-carboxylate (0.382 g, 1.07 mmol, 1.5 eq), PdC12(PPh3)2 (0.014 g, 0.02 mmol, 0.03 eq), Cui (0.0038 g, 0.02 mmol, 0.03 eq), TEA (0.144 g, 1.42 mmol, 2 eq) were dissolved in ACN (2 ml, 0.3 M) and degassed with nitrogen gas. The reaction mixture was heated to 60 °C and stirred for 5 hours. After completion of the reaction, it was extracted with EA, Seong0. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, DCM / MeOH) to obtain the title compound (0.25 g, 65%). Step 4 (d) Synthesis of tert-butyl 4-(2-(4-((5-amino- 2-(pyridin- 4-yl)phenyl)ethynyl)benzamido)ethyl)piperazine-1-carboxylate (tert-butyl 4-(2-(4-((5—amino— 2— (pyridin— 4— yl)phenyl)ethynyl)benzamido)ethyl)piperazine— 1— carboxylate) Tert-butyl 4-(2-(4-((5-nitro- 2-(pyridin- 4 -yl)phenyl)ethynyl)benzamido)ethyl)piperazine-1-carboxylate (tert-butyl 4-(2-(4-((5-nitro— 2— (pyridin— 4— yl)phenyl)ethynyl) )benzamido)ethyl )piperazine— 1— carboxylate, 0.255 g, 0.46 mmol, 1 eq) was dissolved in 1,4 -dioxane: water (H2O) (3:1 (v / v)) (4.6 ml, 0.1 M), then Zn (0.3 g, 4.6 mmol, 10 eq) and NH4CI (0.246 g, 4.6 mmol, 10 eq) were added and stirred at room temperature overnight. It was washed with MeOH and filtered under reduced pressure. The filtrate was concentrated and the next reaction in step 5) was carried out without purification. Step 5 (e) Synthesis of tert-butyl 4-(2-(4-((5-((phenoxycarbonyl)amino)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperazine-1-carboxylate tert-Butyl 4-(2-(4-(((5-amino-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperazine-1-carboxylate, 0.023 g, 0.043 mmol, 1 eq) was dissolved in DMF (0.3 ml, 0.1 M), then pyridine (0.01 g, 0.129 mmol, 3 eq) was added. After cooling to 0 °C, phenyl chloroformate (0.007 g, 0.0473 mmol, 1.1 eq) was added dropwise. After slowly heating to room temperature, it was stirred for 2 hours. After the reaction was completed, it was concentrated, neutralized with 1 N NaOH, and then extracted with EA and water.The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, DCM / MeOH) to obtain the title compound (0.020 g, 72%). Step 6 (f) Synthesis of tert-butyl 4-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperazine-1-carboxylate (tertbutyl 4-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperazine—l— carboxylate) tert-butyl 4-(2-(4-((5-((phenoxycarbonyl)amino)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperazine-1-carboxylate (tert- butyl 4-(2-(4-.

[0483] ((5-(( phenoxycar bony 1 ) am i no ) — 2— ( pyr idin— 4— yl) phenyl) ethynyl) benzamido) ethyl) piperazine— l— carboxylate, 0.030 g, 0.046 mmol, 1 eq) was dissolved in THF (1 ml, 0.05 M), and 2- (pyridin- 3 -yl) ethan- 1-amine (2- (pyridin— 3— yl) ethan- 1— amine, 0.011 g, 0.092 mmol, 2 eq) and TEA (0.014 g, 0.138 mmol, 3 eq) were added and stirred at room temperature overnight. After completion of the reaction, the mixture was concentrated and purified by column chromatography (silica gel, DCM / MeOH) to obtain the title compound (0.021 g, 70%). Step 7 (g) Synthesis of N-(2-(piperazin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (N-(2-(piperazin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide) tert-butyl 4-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide) tert-butyl 4-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperazine-1-carboxylate (tert- Butyl 4-(2-(4- ((5— (3— (2— (pyridin— 3— yl )ethyl )ureido)-2-(pyridin-4- yl)phenyl)ethynyl)benzamido)ethyl)piperazine—l— carboxylate, 0.064 g, 0.094 mmol, leq) was dissolved in DCM (1 ml, 0.1 M), TFA (0.107 g, 0.94 mmol, 10 eq) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the mixture was concentrated and purified by reverse-phase column chromatography (0.The title compound (0.021 g, 40%) was obtained by purification with 1% formic acid in H2O / ACN. P NMR (400 MHz, MeOD) 5 8.63 (d, J= 4.4 Hz, 2H), 8.49 (s, 1H), 8.43 (s, 1H), 7.88 (s, 1H), 7.82 (d, J = 7.9 Hz, 4H), 7.74 (d, J = 4.7 Hz, 2H), 7.45 (d, J = 7.3 Hz, 5H), 3.55 (d, J = 10.7 Hz, 4H), 3.23 (s, 4H), 2.94 (t, .

[0484] J = 6.8 Hz, 2H), 2.79 (s, 4H), 2.69 (t, J = 6.5 Hz, 2H). Example 23: N-Phenyl- 2- ((4- ((2-(piperidin- 1-yl)ethyl)carbamoyl)phenyl)ethynyl)-4- (3- (2-(pyridin- 3 -yl)ethyl)ureido)benzamide (N-pheny 1-2-( (4- ( (2- (piper idin- 1- yl )ethyl ) carbamoyl ) phenyl )ethynyl )-4- (3 -

[0485] Synthesis of (2-(pyridin-3-yl)ethyl)ureido)benzamide) (Synthetic scheme 18) Scheme 18. Reagents and conditions: (a) Aniline, EDC, DMAP, DMF, room temperature (rt), overnight; (b) PdC12(PPh3)2, Cui, TEA, ACN, 60 °C, 5 h; (c) Zn, NH4CI, 1,4-dioxane, water (H2O), room temperature (rt), overnight; (d) phenyl chloroformate, pyridine, THF, 0 °C to room temperature (rt), 2 hrs; (e) 2-(pyridin-3-yl)ethan-1-amine, TEA, THF, room temperature (rt), overnight (overnight) Step 1 (a) Synthesis of 2-iodo-4-nitro-N-phenylbenzamide (2-iodo-4-nitro-N-phenyl benzamide)

[0486] 2-iodo-4-nitrobenzoic acid (0.5 g, 1.71 mmol, 1 eq) was dissolved in DMF (7 ml, 0.25 M), and aniline (0.159 g, 1.71 mmol, 1 eq), EDC (0.392 g, 2.05 mmol, 1.2 eq), and DMAP (0.417 g, 3.42 mmol, 2 eq) were added under nitrogen gas and stirred at room temperature for 16 hours. After completion of the reaction, 5% LiCl aqueous solution (100 ml) was added, stirred, and extracted with DCM. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, Hex / EA) to obtain the title compound (0.341 g, 54%). Step 2 (b) Synthesis of 4-nitro-N-phenyl- 2- ((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)benzamide

[0487] 2-Iodo-4-nitro-N-phenylbenzamide (0.34 g, 0.92 mmol, 1 eq), 4-ethynyl-N-(2-(piperidin-1-yl)ethyl)benzamide (0.353 g, 1.38 mmol, 1.5 eq), PdCl2(PPh3)2 (0.019 g, 0.03 mmol, 0.03 eq), CuI (0.005 g, 0.03 mmol, 0.03 eq), TEA (0.186 g, 1.84 mmol, 2 eq) were dissolved in ACN (4 ml, 0.25 M), and then degassed with nitrogen gas. The reaction mixture was heated to 60 °C and stirred for 5 hours. After completion of the reaction, it was extracted with EA and water. The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.276 g, 60%). Step 3 (c) 4-Amino-N-phenyl-2-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)benzamide (4-amino-N-phenyl-2-((4-((2-

[0488] Synthesis of tert-butyl 4-(2-(4-((5-nitro-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)piperazine-1-carboxylate (tert-butyl 4-(2-(4- ((5— nitro— 2— ( pyr idin— 4— y 1 ) pheny 1 ) ethyny l)benzamido)ethyl)piperazine— 1— carboxylate, 0.1 g, 0.20 mmol , 1 eq) was dissolved in 1,4-dioxane (1,4-dioxane): water (H2O) (3: l(v / v)) (2 ml, 0.1 MH], and Zn (0.13 g, 2 mmol, 10 eq), NH4C1 (0.106 g, 2 mmol, 10 eq) was added and stirred at room temperature overnight. Washed with MeOH and filtered under reduced pressure. The filtrate was concentrated and the next reaction of step 4) was carried out without purification. Step 4 (d) Synthesis of phenyl (4-(phenylcarbamoyl)-3-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)carbamate

[0489] 4-amino-N-phenyl-2-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)benzamide ( 4-amino-N-phenyl-2-((4-((2- (piper idin-l-yl )ethyl )carbamoyl )phenyl )ethynyl )benzamide , 0.093 g, 0.2 mmol , 1 eq) was dissolved in THF (2 ml, 0.1 M) and pyridine (pyr idine, 0.02 g, 0.25 mmol, 1.25 eq) was added. After cooling to 0 °C, phenyl chloroformate (phenyl chloroformate,

[0490] 0.056 g, 0.36 mmol, 1.8 eq) was added dropwise. The mixture was slowly heated to room temperature and stirred for 2 hours. After completion of the reaction, the mixture was concentrated, neutralized with 1 N NaOH, and extracted with EA, 0. The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.052 g, 44%). Step 5 (e) Synthesis of N-phenyl- 2- ((4- ((2- (piperidin- 1-yl)ethyl)carbamoyl)phenyl)ethynyl)- 4- (3- (2- (pyridin- 3-yl)ethyl)ureido)benzamide (N-phenyl- 2- ((4- ((2- (piper idin- 1-yl)ethyl)carbamoyl)phenyl)ethynyl)- 4- (3- (2- (pyr idin- 3- yl)ethyl)ureido)benzamide) phenyl (4- (phenylcarbamoyl)- 3- ((4- ((2- (piperidin- 1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)carbamate ( phenyl (4-( phenyl carbamoyl )-3- ( (4—( (2— (piper idin— 1—yl )ethyl )carbamoyl )phenyl )ethynyl )pheny 1 )carbamat e , 0.052 g, 0.089 mmol, 1 eq) was dissolved in THF (1 ml, 0.1 M), then 2—(pyridin— 3—yl)ethan— 1—amine (2— (pyridin— 3— yl)ethan— 1— amine, 0.022 g, 0.178 mmol, 2 eq) and TEA (0.027 g, 0.267 mmol, 3 eq) were added and stirred at room temperature overnight. After completion of the reaction, the mixture was concentrated and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.033 g, 60%). P NMR (400 MHz, MeOD) 5 8.45 (d, J = 22.7 Hz, 2H) , 8.18 (s, 1H) , 7.96 (d, J = 8.0 Hz, 2H) , 7.92 (s, 1H) , 7.81 (d, J = 7.6 Hz, 1H) , 7.62 (t, J = 7.4 Hz, 2H), 7.56 (d, J = 7.0 Hz, 3H) , 7.46 (d, J = 6.5 Hz, 4H) , 7.33 (d, J = 8.9 Hz, 1H), 6.23 (s, 1H), 3.83 (s, 2H), 3.51 (t, J = 6.5 Hz, 2H), 3.37 (s, 4H), 2.92 (t, J = 6.5 Hz, 2H), 1.93 (s, 4H), 1.72 (s, 2H) Example. 4-( (2- (3 -aminopiperidin- 1-yl )-5- (3- (2- (pyridin- 3-yl)ethyl)ureido)phenyl)ethynyl)- N- (2- (piperidin- 1-yl)ethyl)benzamide (4-((2 -

[0491] ( 3-am i nop i per idi nly 1 )— 5— (3— (2— (pyr idin— 3— yl )ethyl )ureido)phenyl )ethynyl )-

[0492] Synthesis of N-(2-(piperidin-1-yl)ethyl)benzamide) (Synthetic Scheme 19)

[0493] Scheme 19. Reagents and conditions: (a) tert-butyl piperidin-3-yl carbamate, CS2CO3, DMF, 80 °C overnight; (b) PdC12(PPh3)2, Cui, TEA, ACN, 60 °C, 5 h;

[0494] (c) Zn, NH4CI, 1,4-dioxane, water (H2O), room temperature (rt), overnight;

[0495] (d) Phenyl chloroformate, pyridine, THF, 0 °C to room temperature (rt), 2 hours (2 hrs); (e) 2-(Pyridin-3-yl)ethan-1-amine, TEA, THF, 55 °C, overnight; (f) TFA, DCM, room temperature (rt), overnight Step 1 (a) Synthesis of tert-butyl (1-(2-iodo-4-nitrophenyl)piperidin-3-yl)carbamate

[0496] 1-Fluoro-2-iodo-4-nitrobenzene (0.5 g, 1.87 mmol, 1 eq) was dissolved in DMF (3.74 ml, 0.5 M), and then tert-butyl piperidin-3-ylcarbamate (0.412 g, 2.057 mmol, 1.1 eq) and CS2CO3 (0.670 g, 2.057 mmol, 1.1 eq) were added. The mixture was heated at 80 °C for 16 hours with stirring. After completion of the reaction, it was slowly cooled to room temperature, and then extracted with EA and water. The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, Hex / EA) to obtain the title compound (0.614 g, 73%). Step 2 (b) tert-butyl (1-(4-nitro-2-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)piperidin-3-yl)carbamate

[0497] Synthesis of (4-nitro-2-((4-((2-(piper idin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)piper idin-3-yl)carbamate) tert-butyl (1-(2-iodo-4-nitrophenyl)piperidin-3-yl)carbamate (tertbutyl (1—(2— iodo— 4— nitrophenyl)piper idin— 3— yl)carbamate, 0.614 g, 1.37 mmol, 1 eq), 4-ethynyl- N- (2-(piperidin-1-yl)ethyl)benzamide (4-ethynyl- N- (2- (piper i di nl-yl)ethyl)benzamide, 0.527 g, 2.06 mmol, 1.5 eq), PdC12(PPh3)2 (0.028 g, 0.04 mmol, 0.03 eq), Cui (0.008 g, 0.04 mmol, 0.03 eq), and TEA (0.38 g, 2.74 mmol, 2 eq) were dissolved in ACN (13.7 ml, 0.1 M) and degassed with nitrogen gas. The reaction mixture was heated to 60 °C and stirred for 5 hours. After completion of the reaction, the mixture was extracted with EA and C0. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.277 g, 35%).Step 3 (c) Synthesis of tert-butyl (1-(4-amino-2-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)piperidin-3-yl)carbamate tert-butyl (1-(4-nitro-2-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)piperidin-3-yl)carbamate, 0.277 g, 0.48 mmol, 1 eq) was dissolved in 1,4-dioxane: water (3:1) (4.8 ml, 0.1 M HCl), and then Zn (0.314 g, 4.8 mmol, 10 eq) and NH4Cl (0.257 g, 4.8 mmol, 10 eq) were added, and the mixture was stirred at room temperature overnight. It was washed with MeOH and filtered under reduced pressure. The filtrate was concentrated and the next reaction was carried out without purification. Step 4 (d) tert-butyl (1-(4-((phenoxycarbonyl)amino)-2-((4-((2-.

[0498] (Piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)piperidin-3-yl)carbamate (tert-butyl (1-(4-((phenoxycarbonyl)amino)-2-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)piperidin-3-yl)carbamate) Synthesis tert-butyl (1-(4-amino-2-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)piperidin-3-yl)carbamate (tert-butyl (1-(4—amino—2—((4—((2—(piperidin—1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)piperidin—3-yl)carbamate, 0.209 g, 0.38 mmol, 1 eq) was dissolved in THF (3.8 ml, 0.1 M), and then pyridine (0.09 g, 1.14 mmol, 3 eq) was added. After cooling to 0 °C, phenyl chloroformate (0.107 g, 0.68 mmol, 1.8 eq) was added dropwise. After slowly heating to room temperature, the mixture was stirred for 2 hours. After completion of the reaction, it was concentrated, neutralized with 1 N NaOH, and then extracted with EA and water. The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.052 g, 44%). Step 5 (e) tert-butyl (1-(2-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)piperidin-3-yl)carbamate (tert-butyl (l-(2-((4-((2-

[0499] Synthesis of tert-butyl (1- (4-((phenoxycarbonyl)amino)-2- ((4- ((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)piperidin-3-yl)carbamate, 0.1 g, 0.15 mmol, 1 eq) was dissolved in THF (3 ml, 0.05 M), then 2— (pyridin— 3—yl)ethan— 1— amine (2— (pyridin— 3— yl)ethan— 1— amine, 0.054 g, 0.45 mmol, 3 eq) and TEA (0.045 g, 0.45 mmol, 3 eq) were added and stirred at 55°C overnight. After completion of the reaction, the mixture was concentrated and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.070g, 67%) Step 6 (f ) Synthesis of 4-( (2- (3-aminopiperidin-1-yl)- 5- (3- (2-(pyridin-3-yl)ethyl)ureido)phenyl)ethynyl)- N- (2-(piperidin-1-yl)ethyl)benzamide (4- ((2- ( 3-aminopiperidin-1-yl)ethyl)benzamide) tert-butyl (1- (2- ((4- ((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)- 4- (3- (2- (pyridin-3-yl)ethyl)benzamide) -yl)ethyl)ureido)phenyl)piperidin- 3 -yl)carbamate (tert- butyl (1-(2-( (4-( (2-(pi per idin-l-yl )ethyl )carbamoyl )phenyl )ethynyl )-4-(3-(2-.

[0500] (pyridin— 3— yl)ethyl)ureido)phenyl)piper idin— 3— yl)carbamate, 0.030 g, 0.043 mmol, leq) was dissolved in DCM (1 ml, 0.05 M), TFA (0.025 g, 0.215 mmol, 5 eq) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the mixture was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain the title compound (0.0947 g, 37%). P NMR (400 MHz, MeOD) 5 8.46 (s, 2H) , 7.91 (d, J = 6.9 Hz, 2H) , 7.78 (d, J = 7.5 Hz, 1H), 7.65 (d, J = 6.9 Hz, 2H) , 7.59 (s, 1H) , 7.41 (s, 1H), 7.28 (d, J = 8.4 Hz, 1H), 7.01 (d, J = 8.8 Hz, 1H), 3.75 (s, 2H), 3.49 (dd, J = 19.1, 12.4 Hz, 4H), 3.32 (s, 2H), 3.21 (s, 5H) , 2.89 (t, J = 6.7 Hz, 4H) , 2.66 (s, 1H), 2.10 (s, 1H), 1.99 (s, 1H) , 1.85 (s, 5H) , 1.66 (s, 3H) . Example 25: 4- ((3'- (3-aminopyrrolidine-1-carbonyl)-4- (3- (2-(pyridin-3-yl)ethyl)ureido)- [1,1'-biphenyl] -2 - yl)ethynyl)- N- (2-(piperidin-1-yl)ethyl)benzamide ( 4- ( ( 3 ' - ( 3- am i nopyr rol idi ne- 1-carbonyl ) -4 - (3-(2-

[0501] (pyr idin-3-yl )ethyl )ureido)-[l, 1 '-biphenyl ]— 2— yl )ethynyl )— N— (2— (piper idin—

[0502] Synthesis of 1- yl )ethyl )benzamide (Synthetic Reaction Scheme 20)

[0503] Scheme 20. Reagents and conditions: (a) tert-butyl pyrrol idin-3-ylcarbamate, EDC, HOBT, TEA, DCM, room temperature (rt), overnight; (b) Pd(dppf)Cl2, K2C03, 1,4-dioxane, water (H2O), 90 °C, overnight; (c) 10% aqueous p-TSA, NaN02, KI, H2O, ACN, 0 °C, 1 h; (d) PdC12(PPh3)2, Cu, TEA, ACN, 60 °C, 5 h; (e) Zn, NH4Cl, 1,4-dioxane (1,4-dioxane) , water OW), room temperature (rt), overnight; (f) phenyl chloroformate, pyridine, THF, 0 °C to RT, 2 hrs; (g) 2— (pyridin— 3— yl)ethan— 1— amine (2— (pyridin— 3— yl)ethan— 1— amine), TEA, THF, room temperature (rt), overnight; (h) TFA, DCM, room temperature (rt), overnight Step 1 (a) tert-butyl (1-(3-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan- 2-yl)benzoyl)pyrrolidin- 3 -yl)carbamate (tert- butyl (l-(3-(4,4,5,5-tetramethyl- Synthesis of 1,3,2— dioxaborolan— 2— yl )benzoyl )pyrrol idin-3-yl )carbamate)

[0504] 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)benzoic acid (2 g, 8.06 mmol, 1 eq) was dissolved in DCM (80 ml, 0.1 M), and tert-butyl pyrrol idin-3-ylcarbamate (1.8 g, 9.67 mmol, 1.2 eq), EDC (1.853 g, 9.67 mmol, 1.2 eq), HOBT (1.306 g, 9.67 mmol, 1.2 eq), TEA (1.63 g, 16.12 mmol, 2 eq) were added. The mixture was stirred at room temperature overnight. After the reaction was completed, it was extracted with MC and H20. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, EA / Hex) to obtain the title compound (2.1 g, 98%). Step 2 (b) Synthesis of tert-butyl (1-(2'-amino-4'-nitro-[1,1'-biphenyl]-3-carbonyl)pyrrolidin-3-yl)carbamate

[0505] 2-iodo-5-nitroaniline (2-iodo-5-nitroaniline, 0.7 g, 2.65 mmol, 1 eq) was dissolved in 1,4-dioxane: water (H20)(3:l(v / v), 13 ml, 0.14 M), and tert-butyl (1-(3-(4, 4, 5, 5-tetramethyl- 1,3, 2-dioxoborolan- 2-yl)benzoyl)pyrrolidin- 3-yl)carbamate (tert- butyl (l-(3-(4,4,5,5-tetramethyl- 1,3,2— dioxaborolan— 2— yl)benzoyl)pyrrolidin— 3— yl)carbamate, 1.65 g, 3.97 mmol, 1.5 eq), Pd(dppf)C12 (0.387 g, 0.53 mmol, 0.2 eq), K2CO3 (0.805 g, 5.83 mmol, 2.2 eq) was added and degassed with nitrogen gas (N2 gas). The reaction solution was heated to 90 °C and stirred overnight. After completion of the reaction, it was filtered through a celite filter and extracted with EA, 0. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, EA / Hex) to obtain the title compound (1.1 g, 97%). Step 3 (c) Synthesis of tert-butyl (1-(2'-iodo-4'-nitro-[1,1'-biphenyl]-3-carbonyl)pyrrolidin-3-yl)carbamate (tert-butyl (1-(2'-iodo-4'-nitro-[1,1'-biphenyl]-3-carbonyl)pyrrolidin-3-yl)carbamate) 1-amino-4'-nitro-[1,1'-biphenyl]-3-carbonyl)pyrrolidin-3-yl)carbamate, 0.6 g, 1.4 mmol, 1 eq) was dissolved in ACN (14 ml, 0.1 M), and then an aqueous solution of 10% p-TSA (1.687 g, 9.8 mmol, 7 eq) was added. An aqueous solution of 0.1 M NaNO2 (0.106 g, 1.54 mmol, 1.1 eq) was added dropwise over 0.5 h. 0.1 M NaI (0.419 g, 2.8 mmol, 2 eq) was added to the reaction solution at 0 °C in one portion, and the mixture was stirred for 0.5 h. After completion of the reaction, water (10 ml) was added to the reaction solution, and the mixture was extracted with EA. The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, EA / Hex) to obtain the title compound (0.671 g, 89%). Step 4 (d) tert-Butyl (1-(4 1tert-Butyl (1-(4'-nitro-2'-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)-[1,1'-biphenyl]-3-carbonyl)pyrrolidin-3-yl)carbamate (tert-butyl (1-(2'-iodo-4'-nitro-[1,1'-biphenyl]-3-carbonyl)pyrrolidin-3-yl)carbamate, 0.3 g, 0.56 mmol, 1 eq), 4-ethynyl-N-(2-(piperidin-1-yl)ethyl)benzamide (4-ethynyl-N-(2-(piperidin-1-yl)ethyl)benzamide, 0.215 g, 0.84 mmol, 1.5 eq), PdCl2(PPh3)2 (0.012 g, 0.017 mmol, 0.03 eq), CuI (0.003 g, 0.017 mmol, 0.03 eq), and TEA (0.113 g, 1.12 mmol, 2 eq) were dissolved in ACN (5.6 ml, 0.1 M) and degassed with nitrogen gas. The reaction mixture was heated to 60 °C and stirred for 5 hours. After completion of the reaction, the mixture was extracted with EA and water. The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.316 g, 85%).Step 5 (e) Synthesis of tert-butyl (1-(4'-amino- 2'- ((4- ((2- (piperidin- 1-yl)ethyl)carbamoyl)phenyl)ethynyl)- [1,1'-biphenyl] -3 -carbonyl)pyrrolidin- 3 -yl)carbamate (tert- butyl (1-(4'-amino- 2'- ((4- ((2- (piperidin- 1- yl)ethyl)carbamoyl)phenyl)ethynyl)— [1,1' -biphenyl]— 3 -carbonyl)pyrrolidin- 3 -yl)carbamate) (1-(4'-nitro-2'-((4-((2- (piperidin- 1- yl )ethyl )carbamoyl )phenyl )ethynyl )— [1 , 1 ' -bi pheny 1 ] -3~carbony 1 )pyrrol idin— 3— yDcarbamate, 0.2 g, 0.3 mmol , 1 eq) was dissolved in 1 , 4-dioxane ( 1 , 4-dioxane ) : water (H2O) (3 : l(v / v)) (3 ml, 0.1 MH], and then Zn (0.196 g, 3 mmol, 10 eq) and NH4C1 (0.16 g, 3 mmol, 10 eq) were added and stirred at room temperature overnight. Washed with MeOH and filtered under reduced pressure. The filtrate was concentrated and used for the next reaction of step 6) without purification. Step 6 (f) Tert-butyl (1-(4'-((phenoxycarbonyl)amino)- 2'- ((4-((2-.

[0506] (piperidin-1-yl)ethyl)carbamoyl)phenol)ethynyl)-[1,1'-biphenyl]-3-carbonyl)pyrrolidin-3-yl)carbamate (tert-butyl (1-(4'-((phenoxycarbony 1)amino)-2 1 - ( (4-((2-

[0507] Synthesis of tert-butyl (1-(4'-amino-2'-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)-[l,1'-biphenyl]-3-carbonyl)pyrrolidin-3-yl)carbamate) tert-butyl (1-(4'-amino-2'-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)-[1,1'-biphenyl]-3-carbonyl)pyrrolidin-3-yl)carbamate, 0.19 g, 0.3 mmol, 1 eq) was dissolved in THF (3 ml, 0.1 M), and pyridine (0.07 g, 0.9 mmol, 3 eq) was added. After cooling to 0 °C, phenyl chloroformate (0.084 g, 0.54 mmol, 1.8 eq) was added dropwise. The mixture was slowly heated to room temperature and stirred for 2 hours. After completion of the reaction, the mixture was concentrated, neutralized with 1 N NaOH, and extracted with EA, 0. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.083 g, 33%). Step 7 (g) tert-Butyl (1- (2 1 - ( (4- ( (2- (piperidin- 1-yl)ethyl)carbamoyl)phenyl)ethynyl)- 4'- (3- (2- (pyridin- 3 -yl)ethyl)ureido)- [1,1'-biphenyl] -3 -carbonyl)pyrrolidin- 3 -yl)carbamate (tert- butyl (l- (2'- ((4- ((2-

[0508] Synthesis of tert-butyl (1- (4'- ((phenoxycarbonyl)amino)- 2'- ((4- ((2- (piperidin- 1-yl)ethyl)carbamoyl)phenyl)ethynyl)- 4'- (3- (2- (pyridin- 3 -yl)ethyl)ureido)- [ 1,1'-biphenyl]- 3-carbony l)pyrrolidin- 3-yl)carbamate) tert-butyl (1- (4'- ((phenoxycarbonyl)amino)- 2'- ((4- ((2- (piperidin- 1-yl)ethyl)carbamoyl)phenol)ethynyl)- [1,1'-biphenyl] -3 -carbonyl)pyrrolidin- 3-yl)carbamate

[0509] (piper idin-l-yl)ethyl )carbamoyl )phenyl )ethynyl )— [1 , 1 ' -biphenyl ] -3- carbonyl )pyrrol idin-3-yl )carbamate, 0.083 g, 0.1 mmol , 1 eq) was dissolved in THF (1 ml, 0.1 M), then 2-(pyridin-3-yl)ethan-1-amine (2-(pyridin-3- yl)ethan-1-amine, 0.024 g, 0.2 mmol, 2 eq) and TEA (0.030 g, 0.3 mmol, 3 eq) were added and stirred at 55 °C overnight. After completion of the reaction, the mixture was concentrated and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.049 g, 62%). Step 8 (h) 4-((3 '-(3-aminopyrrolidine-1-carbonyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)-[1,1'-biphenyl]-2-yl)ethynyl)- N-(2-(piperidin-1-yl)ethyl)benzamide (4-((3 '-(3-aminopyrrol idine-l-carbonyl)-4-(3-(2-

[0510] Synthesis of tert-butyl (1-(2'-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)- (pyridin-3-yl)ethyl)ureido)-[ 1,1 '-biphenyl] -2-yl )ethynyl )-N-(2-(piperidin- l-yl )ethyl )benzamide)

[0511] 4'- (3- (2- (pyridin- 3 -yl)ethyl)ureido)- [1,1'-biphenyl] -3 -carbonyl)pyrrolidin- 3-yl)carbamate (tert- butyl (1- (2 '-( (4-( (2- (pi per idin-l- yl )ethyl )carbamoyl )phenyl )ethynyl )— 4' — (3— (2— (pyr idin— 3— yl )ethyl )ureido)- [1,1' -biphenyl ]-3-carbony 1 )pyrrol idin-3-yl )carbamate, 0.049 g, 0.062 mmol , leq) was dissolved in DCM (1 ml, 0.05 M), TFA (0.035 g, 0.31 mmol, 5 eq) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the product was concentrated and purified by reverse phase column chromatography (0.1% formic acid in H2O / ACN) to obtain the title compound (0.014 g, 33%). P NMR (400 MHz, MeOD) 5 8.39 (s, 2H) , 8.28 (s, 1H) , 7.94 (d, J = 5.5 Hz, 2H), 7.85 (d, J = 7.8 Hz, 1H) , 7.79 — 7.71 (m, 1H) , 7.65 — 7.45 (m, 6H), 7.38 (s, 1H), 7.33 (t, J = 7.6 Hz, 1H), 7.30 - 7.22 (m, 1H), 4.05 - 3.88 (m, 2H), 3.78 (d, J = 22.7 Hz, 5H), 3.63 (s, 1H) , 3.53 — 3.33 (m, 6H) , 3.21 (dd, J = 14.6, 7.3 Hz, 1H), 2.87 — 2.77 (m, 2H) , 2.66 (s, 1H) , 2.40 (s, 1H) , 2.16 (s, 1H), 1.95 - 1.86 (m, 4H) , 1.69 (s, 2H) , 1.33 (dd, J = 16.2, 8.9 Hz, 1H) .Example 26: 1-(2-((4-fluorophenyl)ethynyl)-4'-(piperazin-1-yl)-[1,1'-biphenyl]-4-yl)-3-(imidazo[1,2-a]pyridin-7-ylmethyl)urea (1-(2-((4-f luorophenyl)ethynyl)— 4'—(piperazin— 1—yl)-[l, 1'-biphenyl] -4-yl)-3-.

[0512] Synthesis of (imidazo[l,2- a]pyridin-7-ylmethyl)urea) (Synthetic reaction scheme 21)

[0513] Scheme 21. Reagents and conditions: (a) phenyl chloroformate, pyridine, THF, 0 °C to room temperature (rt); (b) imidazo[l,2-a]pyridin-7-ylmethanamine IC1, TEA, THF, reflux, overnight; (c)

[0514] Pd(dppf)C12, K2CO3, 1,4-dioxane, water (H2O), 90 °C overnight;

[0515] (d) TFA, DCM, room temperature (rt), overnight Step 1 (a) Phenyl (3-((4-fluorophenyl)ethynyl)-4-iodophenyl)carbamate

[0516] Synthesis of (phenyl (3- ((4- fluorophenyl ) ethynyl )-4- iodophenyl ) carbamate)

[0517] 3-((4-Fluorophenyl)ethynyl)-4-iodoaniline (3-((4-fluorophenyl)ethynyl)—4—iodoaniline, 1 g, 2.97 mmol, 1 eq) was dissolved in THF (10 ml, 0.3 M), and then pyridine (0.329 g, 41.58 mmol, 14 eq) and phenyl chloroformate (0.558 g, 3.564 mmol, 1.2 eq) were added dropwise with stirring at 0 °C. After completion of the reaction, it was concentrated and extracted with EA and water (Cheng 0). The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, EA / Hex) to obtain the title compound (1.3 g, 96%). Step 2 (b) 1-(3-((4-Fluorophenyl)ethynyl)-4-iodophenyl)-3-

[0518] It should be noted that there seems to be an unclear or incorrect expression "水 (成0)" in the original text which might need further clarification for a more accurate translation.Synthesis of (imidazo[1,2-a]pyridin-7-ylmethyl)urea (1-(3-((4-f luorophenyl)ethynyl 1)-4-iodophenyl )-3-( imidazot 1 , 2-a]pyridin-7-y Imethy 1 )urea) Phenyl (3-((4-fluorophenyl)ethynyl)-4-iodophenyl)carbamate (phenyl (3-((4-f luorophenyl )ethynyl )-4-iodopheny 1 ) carbamate , 1 g, 2.18 mmol , 1 eq) was dissolved in THF (21.8 ml, 0.1 M), and imidazo[1,2-a]pyridin-7-ylmethanamine* HC1 salt (imidazo[l,2— a]pyridin— 7— After adding ylmethanamine IC1 salt (0.8 g, 4.36 mmol, 2 eq), TEA (0.662 g, 6.54 mmol, 3 eq), the mixture was heated to reflux and stirred overnight. After completion of the reaction, the mixture was concentrated, crystallized with Hex and MC, and the crystals were washed three times with 0 and filtered to obtain the title compound (0.85 g, 76%). Step 3 (c) tert-Butyl 4- (2'- ((4-fluorophenyl)ethynyl)-4'- (3-

[0519] Synthesis of (tert-butyl 4-(2'-((4-fluorophenyl)ethynyl)-4'-(3-(imidazo[l,2- a]pyridin— 7— y Imethy l)urei do)— [1,1'— biphenyl]— 4— yl)piperazine— 1— carboxylate)

[0520] 1-(3-((4-fluorophenyl)ethynyl)-4-iodophenyl)-3-(imidazo[1,2-a]pyridin—7—ylmethyl)urea 0} (1— (3—((4— fluorophenyl)ethynyl)— 4— iodophenyl)— 3—

[0521] (Imidazo[l,2— a]pyridin— 7— ylmethyl)urea, 0.1 g, 0.19 mmol, 1 eq) was dissolved in 1,4-dioxane: water (H20)(3: l(v / v), 0.8 ml, 0.25 M), and (4-(4-(tert-butoxycarbonyl)piperazin— 1—yl)phenyl)boronic acid ((4-(4-(tert- butoxycarbonyl )piperaz in— 1—yl )phenyl )boronic acid, 0.11 g, 0.38 mmol , 2 eq), Pd(dppf )C12 (0.027 g, 0.038 mmol , 0.2 eq), K2CO3 (0.058 g, 0.418 mmol , 2.2 eq) was added and degassed with nitrogen gas. The reaction solution was heated to 90 °C and stirred overnight. After completion of the reaction, it was filtered through a celite filter and extracted with EA, 0. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.024 g, 19%).Step 4 (d) Synthesis of tert-butyl 4-(2'-((4-fluorophenyl)ethynyl)- 4'-(piperazin-1-yl)- [1,1'-biphenyl] -4 -yl)- 3-(imidazo[1,2-a]pyridin- 7 -ylmethyl)urea -1- Carboxylate (tert- butyl 4-(2'-((4-fluorophenyl)ethynyl)-4'-(3-(imidazo[l,2- a]pyridin— 7—y lmethyl)urei do)— [1,1'— biphenyl]— 4— yl )pi per azine— 1— carboxylate, 0.024 g, 0.037 mmol, leq) was dissolved in DCM (1 ml, 0.03 M), TFA (0.021 g, 0.185 mmol, 5 eq) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the residue was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain the title compound (0.007 g, 35%). P NMR (400 MHz, MeOD) 5 8.42 (s, 1H) , 7.83 (s, 1H) , 7.72 (s, 1H) , 7.61 - 7.49 (m, 4H) , 7.45 - 7.31 (m, 4H) , 7.08 (d, J = 6.9 Hz, 4H) , 6.97 (d, J = 6.6 Hz, 1H), 4.51 (s, 2H) , 3.62 (s, 4H) , 3.21 (s, 4H) .Example 27: Synthesis of 1-(imidazo[l,2-a]pyridin-7-ylmethyl)-3-(3-((4-(piperidin-4-yl)phenyl)ethynyl)-4-(pyridin-4-yl)phenyl)urea (1-(imidazo[l,2-a]pyridin-7-ylmethyl)-3-(3-((4-(piper idin-4- yl)phenyl)ethynyl)-4-(pyr idin-4- yl)phenyl)urea) (Synthetic Scheme 22).

[0522] Scheme 22. Reagents and conditions: (a) trimethylsilylacetylene, PdC12(PPh3)2, Cui, TEA, toluene, 100 °C for 12 h; (b) K2C03, MeOH, room temperature (rt), 2 h; (c) PdC12(PPh3)2, Cui, TEA, ACN, 60 °C for 1 h; (d) phenyl chloroformate, pyridine, THF, 0 °C to room temperature (rt); (e) imidazo[1,2-a]pyridin-7-ylmethanamine, TEA, THF, 80 °C overnight; (f) TFA, DCM, room temperature (rt), overnight, Step 1 (a) Synthesis of tert-butyl 4-(4-((trimethylsilyl)ethynyl)phenyl)piperidine-1-carboxylate tert-Butyl 4-(4-bromophenyl)piperidine-1-carboxylate (0.5 g, 1.47 mmol, 1 eq) was dissolved in toluene (7.4 ml, 0.2 M), and then trimethylsilylacetylene (0.216 g, 2.2 mmol, 1.5 eq), PdCl2(PPh3)2 (0.103 g, 0.147 mmol, 0.1 eq), Cui (0.027 g, 0.147 mmol, 0.1 eq), TEA (0.594 g, 5.88 mmol, 4 eq) were added, and then degassed with nitrogen gas (N2 gas).The reaction mixture was heated to 100 °C and stirred for 12 hours. After completion of the reaction, it was extracted with DCM, and the organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (Silica gel, EA / Hex) to obtain the title compound (0.196 g, 37%). Step 2 (b) Synthesis of tert-butyl 4-(4-ethynylphenyl)piperidine-1-carboxylate Tert-butyl 4-(4-((trimethylsilyl)ethynyl)phenyl)piperidine-1-carboxylate.

[0523] (tert-butyl 4-(4-( (trimethylsi lyl )ethynyl )phenyl )piperidine—l— carboxylate, 0.196 g, 0.55 mmol, 1 eq) was dissolved in MeOH (5.5 ml, 0.1 M), and K2C03 (0.38 g, 2.75 mmol, 5 eq) was added and stirred at room temperature for 2 hours. After completion of the reaction, the mixture was filtered, concentrated, and purified by column chromatography (silica gel, EA / Hex) to obtain the title compound (0.156 g, 100%). Step 3 (c) Synthesis of tert-butyl 4-(4-((5-amino-2-(pyridin-4-yl)phenyl)ethynyl)phenyl)piperidine-1-carboxylate

[0524] 3-iodo— 4-(pyridin-4-yl)aniline (3-i odo-4-(pyr idi n-4-yl)ani line,

[0525] 0.1 g, 0.37 mmol, 1 eq), tert-butyl 4-(4-ethynylphenyl)piperidine-1-carboxylate (0.157 g, 0.55 mmol, 1.5 eq), PdCl2(PPh3)2 (0.007 g, 0.01 mmol, 0.03 eq), CuI (0.002 g, 0.01 mmol, 0.03 eq), TEA (0.075 g, 0.74 mmol, 2 eq) were dissolved in ACN (3.7 ml, 0.1 M), and then degassed with nitrogen gas. The reaction mixture was heated to 60 °C and stirred for 1 hour. After completion of the reaction, it was extracted with EA and H2O. The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.075 g, 43%). Step 4 (d) tert-butyl 4-(4-((5-((phenoxycarbonyl)amino)-2-(pyridin-4-yl)phenyl)ethynyl)phenyl)piperidine-1-carboxylate (tert-butyl 4-(4-((5-

[0526] Synthesis of ( (phenoxycarbonyl )amino)— 2— (pyridin— 4— yl )phenyl )ethynyl )phenyl )piper idine- 1-carboxylate Tert-butyl 4- (4- ((5-amino-2-(pyridin- 4 -yl)phenyl)ethynyl)phenyl)piperidine- 1-carboxylate (tert- butyl 4- ( 4- ( ( 5-amino-2- (pyridin- 4 - yl)phenyl)ethynyl)phenyl)piperidine—l— carboxylate, 0.075 g, 0.16 mmol , 1 eq) was dissolved in THF (1.6 ml, 0.1 M), and then pyridine (pyr idine, 0.177 g, 2.24 mmol , 14 eq), phenyl Chloroformate (phenyl chloroformate 0.030 g, 0.192 mmol,

[0527] 1.2 eq) was added dropwise and stirred. After completion of the reaction, the mixture was concentrated and extracted with EA, C0. The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.06 g, 63%). Step 5 (e) tert-butyl 4- (4- ((5- (3- (imidazo [1,2- a]pyridin- 7-ylmethyl)ureido)- 2- (pyridin- 4 -yl)phenyl)ethynyl)phenyl)piperidine- 1-carboxylate (tert-butyl 4- (4- ((5- (3- (imidazo [l,2- a]pyr idin- 7- ylmethyl )ureido)- 2-

[0528] Synthesis of tert-butyl 4-(4-((5-((phenoxycarbonyl)amino)-2-(pyridin-4-yl)phenyl)ethynyl)phenyl)piperidine—l— carboxylate) (tert- butyl 4-(4-((5-

[0529] ((phenoxycarbonyl)amino)— 2— (pyridin— 4— yl)phenyl)ethynyl)phenyl)piperidine— 1-carboxylate, 0.060 g, 0.1 mmol, 1 eq) was dissolved in THF (1 ml, 0.1 M), and then imidazo[l,2- a]pyridin- 7-ylmethanamine IC1 salt (0.037 g, 0.2 mmol, 2 eq) and TEA (0.030 g, 0.3 mmol, 3 eq) were added. The mixture was heated to reflux and stirred overnight. After completion of the reaction, the mixture was concentrated and the next reaction of step 6) was performed without purification. Step 6 (f) Synthesis of tert-butyl 4-(4-((5-(3-(imidazo[1,2-a]pyridin-7-ylmethyl)-3-(3-((4-(piperidin-4-yl)phenyl)ethynyl)-4-(pyridin-4-yl)phenyl)urea (1-(imidazo[l,2-a]pyridin-7-ylmethyl)—3—(3—((4—(piper idin—4— yl)phenyl)ethynyl)—4—(pyr idin—4— yl)phenyl)urea) tert-butyl 4-(4-((5-(3-(imidazo[1,2-a]pyridin-7-ylmethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)phenyl)piperidine-1-carboxylate (tert-butyl 4-(4-((5- (3— (imidazo[l,2— a]pyridin— 7— ylmethyl)ureido)— 2— (pyridin— 4— yl)phenyl)ethynyl)phenyl)piperidine—l— carboxylate, 0.063 g, 0.1 mmol, leq) was dissolved in DCM (1 ml, 0.1 M), TFA (0.057 g, 0.5 mmol, 5 eq) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the mixture was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain the title compound (0.021 g, 40%) was obtained. P NMR (400 MHz, MeOD) 5 8.60 (d, J = 5.4 Hz, 2H), 8.56 (d, J = 6.9.

[0530] Hz, 1H), 8.21 s, 1H), 7.97 (s, 1H), 7.88 (d, J = 1.7 Hz, 1H), 7.76 - 7.71

[0531] (m, 3H), 7.68 s, 1H), 7.53 (dd, J = 8.4, 1.9 Hz, 1H), 7.44 (d, J = 8.5 Hz,

[0532] 1H), 7.32 (d, J = 8.1 Hz, 2H), 7.27 (d, J = 8.2 Hz, 2H), 7.20 (d, J = 6.8

[0533] Hz, 1H), 4.58 s, 2H), 3.51 (d, J = 12.6 Hz, 2H), 3.14 (t, J = 11.8 Hz, 2H),

[0534] 2.92 (t, J = 12.2 Hz, 1H), 2.07 (d, J = 13.6 Hz, 2H), 1.92 (t, J = 11.8 Hz,

[0535] 2H); MS MH + 527.25 Example 28: Synthesis of N-(2-(2,8-diazaspiro[4.5]decan-8-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (Synthetic Scheme 23)

[0536] Scheme 23. Reagents and conditions: (a) HATU, DIPEA, DMF, room temperature (rt), overnight; (b) PdC12(PPh3)2, Cui, TEA, ACN, 60 °C 5 h; (c) TFA, DCM, room temperature (rt), overnight. Step 1 (a) Synthesis of tert-butyl 8-(2-(4-ethynylbenzamido)ethyl)-2,8-diazospiro[4.5]decane-2-carboxylate

[0537] 4-ethynyl benzoic acid (0.05 g, 0.34 mmol, 1 eq) in DMF

[0538] (Dissolved in (3.4 ml, 0.1 M), HATU (0.193 g, 0.51 mmol, 1.5 eq) was added and stirred at room temperature for 10 minutes. To the reaction solution, tert-butyl 8-(2-aminoethyl)-2,8-diazaspiro[4.5]decane-2-carboxylate (0.149 g, 0.51 mmol, 1.5 eq) and DIPEA (0.131 g, 1.02 mmol, 3 eq) were added and stirred at room temperature overnight. After completion of the reaction, it was extracted with EA, saturated brine. The organic layer was dried over MgSO4, filtered, concentrated and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.118 g, 84%). Step 2 (b) Synthesis of tert-butyl 8-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)-2,8-diazaspiro[4.5]decane-2-carboxylate

[0539] 1-(3-iodo-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea, 0.05 g, 0.11 mmol, 1 eq), tert-butyl 8-(2-(4-ethynylbenzamido)ethyl)-2,8-diazospiro[4.5]decane-2-carboxylate, 0.068 g, 0.165 mmol, 1.5 eq), PdC12(PPhs)2 (0.002 g, 0.0033 mmol, 0.03 eq), Cui (0.00063 g, 0.0033 mmol, 0.03 eq), TEA (0.022 g, 0.22 mmol, 2 eq) were dissolved in ACN (1.1 ml, 0.1 M) and degassed with nitrogen gas (N2O). The reaction mixture was heated to 60 °C and stirred for 5 hours. After completion of the reaction, it was extracted with DCM: MeOH (9: l(v / v)), water (H2O). The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.052 g,

[0540] 65%) was obtained. Step 3 (c) N-(2-(2,8-diazaspiro[4.5]decan-8-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (N-(2-

[0541] Synthesis of tert-butyl 8- (2- (4- ( (5- (3- (2- (pyridin-3- yl)ethyl)ureido)-2- (pyridin-4-yl)phenyl)ethynyl)benzamide) tert-butyl 8- (2- (4- ( (5- (3- (2- (pyridin-3- yl)ethyl)ureido)-2- (pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)- 2,8-diazaspiro [4.5]decane- 2 -carboxylate (tert-butyl 8- (2- (4- ((5- (3- (2- (pyridin-3- yl)ethyl)ureido)-2- (pyridin-4- yl)) phenyl )ethynyl )benzamido)ethyl )-2, 8-di azaspiro [4.5] decane-2-carboxy late, 0.052 g, 0.071 mmol, leq) was dissolved in DCM (1 ml, 0.07 M), TFA (0.04 g, 0.36 mmol, 5 eq) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the residue was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain the title compound (0.015 g, 34%). P NMR (400 MHz, MeOD) 5 8.59 (d, J = 3.9 Hz, 2H) , 8.48 (s, 1H) , 8.40 (s, 1H), 8.33 (s, 1H), 7.84 (dd, J = 21.1, 10.8 Hz, 4H) , 7.70 (d, J = 4.5 Hz, 2H), 7.47 (d, J = 8.5 Hz, 1H) , 7.42 (d, J = 7.1 Hz, 3H) , 3.73 (s, 2H), 3.51 (t, J = 6.5 Hz, 2H), 3.41 (s, 2H) , 3.15 (d, J = 29.0 Hz, 8H) , 2.92 (t, J = 6.5 Hz, 2H), 2.05 — 1.79 (m, 6H); MS MH +628.5 Example 29: N-(2-(3-aminopyrrolidin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (N-(2-(3-aminopyrrol idin-l-yl)ethyl)— 4— ( (5— (3— (2— (pyr idin— 3— yl)ethyl)ureido)—2—

[0542] Synthesis of ( py ridi n-4-y 1 ) pheny 1 ) et hyny 1 ) benz amide) (Synthetic reaction scheme 24) Scheme 24. Reagents and conditions: (a) HATU, DIPEA,

[0543] DMF, room temperature (rt), overnight; (b) PdCl2(PPh3)2, Cui, TEA, ACN, 60 °C for 5 h: (c) TFA, DCM, room temperature (rt), overnight Step 1 (a) Synthesis of tert-butyl (1-(2-(4-ethynylbenzamido)ethyl)pyrrolidin-3-yl)carbamate

[0544] 4-Ethynylbenzoic acid (0.05 g, 0.34 mmol, 1 eq) was dissolved in DMF (3.4 ml, 0.1 M), then HATU (0.193 g, 0.51 mmol, 1.5 eq) was added and the mixture was stirred at room temperature for 10 minutes. Tert-butyl (1-(2-aminoethyl)pyrrolidin-3-yl)carbamate (0.117 g, 0.51 mmol, 1.5 eq) and DIPEA (0.131 g, 1.02 mmol, 3 eq) were added to the reaction solution and the mixture was stirred at room temperature overnight. After completion of the reaction, the mixture was extracted with EA and water. The organic layer was dried over MgSO₄, filtered, concentrated and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.063 g, 26%). Step 2 (b) Synthesis of tert-butyl (1-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)ethyl)pyrrolidin-3-yl)carbamate

[0545] 1-(3-Iodo-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (0.053 g,

[0546] tert-butyl (1-(2-(4-ethynylbenzamido)ethyl)pyrrolidin-3-yl)carbamate (0.12 mmol, 1 eq), tert-butyl (1-(2-(4-ethynylbenzamido)ethyl)pyrrolidin-3-yl)carbamate (0.063 g, 0.176 mmol, 1.5 eq), PdC12(PPh3)2 (0.003 g, 0.0036 mmol, 0.03 eq), Cui (0.00069 g, 0.0036 mmol, 0.03 eq), TEA (0.024 g, 0.24 mmol, 2 eq) were dissolved in ACN (1.2 ml, 0.1 M), and then degassed with nitrogen gas (G gas). The reactants were The mixture was heated to 60 °C and stirred for 5 hours. After completion of the reaction, the mixture was extracted with DCM: MeOH (9: l(v / v)) and water (0). The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.036 g, 44%).Step 3 (c) Synthesis of N-(2-(3-aminopyrrolidin-1-yl)ethyl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (N-(2-(3-aminopyrrol idin-l-yl )ethyl )— 4— ( (5— (3— (2— (pyridin— 3— yl )ethyl )ureido)— 2— (pyridin-4-yl )phenyl )ethynyl )benzamide) tert-butyl (1-(2-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)- 2-(pyridin-4-yl)phenyl)ethynyl)benzamide) - (tert-butyl (1- (2— (4— ((5— (3— (2— (pyridin— 3— yl )ethyl )ureido)-2-(pyridin-4- yl )phenyl )ethynyl )benzamido)ethyl )pyrrol idin-3-yl )carbamate, 0.036 g, 0.053 mmol, leq) was dissolved in DCM (1 ml, 0.05 M), TFA (0.06 g, 0.53 mmol, 10 eq) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the residue was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain the title compound (0.020 g, 66%). P NMR (400 MHz, MeOD) 5 8.61 (d, J = 5.9 Hz, 1H) , 8.48 (s, 1H) , 8.41 (d, J = 4.4 Hz, 1H), 8.12 (s, 2H) , 7.85 (dt, J = 22.1, 7.8 Hz, 4H) , 7.73 (d,.

[0547] J = 5.9 Hz, 1H), 7.64 (d, J = 8.1 Hz, 1H) , 7.46 — 7.39 (m, 4H) , 3.83 (s, 1H) ,

[0548] 3.65 - 3.55 (m, 2H) , 3.51 (t, J = 7.0 Hz, 2H), 3.19 (s, 1H) , 3.12 - 3.01 (m,

[0549] 1H), 2.96 - 2.84 (m, 5H), 2.62 (dd, J = 17.1, 9.1 Hz, 2H), 2.38 (s, 1H),

[0550] 1.90 (s, 1H). Example 30: Synthesis of 1-(3-((3-aminophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((3-aminophenyl)ethynyl 1)-4-(pyri din-Aryl)phenyl)-3-(2-(pyri din-3-yl)ethyl)urea) (Synthetic Reaction Scheme 25)

[0551] 1 Reaction Scheme 25. Reagents and conditions: (a) 3-ethynylaniline, PdC12(PPh3)2, Cui, TEA, ACN, 80 °C for 3 h

[0552] 1-(3-iodo-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (0.050 g, 0.11 mmol, 1 eq), 3-ethynylaniline (0.019 g, 0.165 mmol, 1.5 eq), PdCl2(PPh3)2 (0.007 g, 0.011 mmol, 0.1 eq), CuI (0.002 g, 0.011 mmol, 0.1 eq), and TEA (0.044 g, 0.44 mmol, 4 eq) were dissolved in ACN (1.1 ml, 0.1 M) and degassed with nitrogen gas. The reaction mixture was heated to 80 °C and stirred for 3 h. After completion of the reaction, it was extracted with DCM:MeOH (9:1 (v / v)). The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (15 mg, %). 1H NMR (400 MHz, DMSO) δ 8.79 (s, 1H), 8.66 (s, 2H), 8.46 (d, J = 13.7 Hz, 2H), 7.83 (s, 1H), 7.68 (d, J = 7.8 Hz, 1H), 7.64 (d, J = 5.4 Hz, 2H), 7.43 (d, J = 1.1 Hz, 2H), 7.35 (dd, J = 7.6, 4.8 Hz, 1H), 7.02 (dd, J = 8.4, 7.5 Hz, 1H), 6.58 (dd, J = 8.6, 1.1 Hz, 2H), 6.53 (d, J = 7.5 Hz, 1H), 6.31 (t, J = 5.7 Hz, 1H), 5.26 (s, 2H), 3.39 (dd, J = 12.8, 6.8 Hz, 2H), 2.80 (t, J = 7.0 Hz, 2H).Example 31: Synthesis of 4-((2-(1-(cyclohexylamino)-1-oxopropan-2-yl)-5-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)ethynyl)-N-(2-(piperidin-1-yl)ethyl)benzamide (4— ((2— (1— ( cyclohexylamino) — 1— oxopr opan-2-yl) — 5— ( 3— ( 2— ( pyr i di n— 3— yl )ethyl )ureido)phenyl )ethynyl )— N— (2— (piper idin— 1— yl )ethyl )benzamide) (Synthetic Scheme 26).

[0553] Reaction Scheme 26 (a) SOCI2, MeOH, room temperature (rt); (b) NaH, Mel, DMF, 4 h (4 h); (c) NaOH, H20, MeOH, reflux (ref lux), 4 h (4 h); (d) TBTU, TEA, cyclohexanamine, DCM, 24 h (24 h); (e) PdtPPhsh, Cui, TEA, 4 —ethynyl— N— (2—(piperidin-1—yl)ethyl)benz 0 {4-ethynyl-N-(2-(piper idin-1- yl)ethyl)benzamide), ACN, room temperature (rt), 1 h (Ih); (f) Fe, NH4C1, EtOH, H2O, 90 °C 2 h (2h); (g) phenyl chloroformate, pyridine, THF, room temperature (RT), 2 h (2h); (h) 2-(pyridin-3-yl)ethan-1-amine, TEA, THF, reflux, 24 h (24h) Step 1 (a) methyl 2-(2-iodo-4-nitrophenyl)acetate )acetate) synthesis

[0554] 2- (2-iodo-4-nitrophenyl)acetic acid (2- (2- iodo-4-nitrophenyl)acetic acid, 1 g, 3.26 mmol, 1 eq) was dissolved in MeOH (0.25 MH], the reaction mixture was cooled with ice, and S0C12 (7.8 ml) was slowly added dropwise. The mixture was stirred at room temperature, and when the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain the title compound. Step 2 (b) Synthesis of methyl 2- (2- iodo-4-nitrophenyl)propanoate Methyl 2- (2-iodo-4-nitrophenyl)acetate (3.25 mmol) was dissolved in DMF (6 ml ) was dissolved in 0.24 ml of water and Mel (0.24 ml, 1.2 eq) was added. The reaction solution was cooled with ice and 60% NaH (0.14 g, 3.58 mmol, 1.1 eq) was added in portions and stirred for 4 hours. Water was added to the reaction mixture and extracted with distilled water. The organic layer was washed with a saturated aqueous ammonium chloride solution, dried over magnesium sulfate, and concentrated under reduced pressure to obtain the title target compound (1.2 g). Step 3 (c) Synthesis of 2-(2-iodo-4-nitrophenyl)propanoic acid Methyl 2-(2-iodo-4-nitrophenyl)propanoate (methyl 2-(2-iodo-4-nitrophenyl)propanoate, 0.36 g, 2.98 mmol, 1 eq), A mixture of NaOH (0.3 g, 7.5 mmol, 2.5 eq) dissolved in water (0.2 M) and MeOH (0.1 M) was stirred at 60°C for 2-4 h. The reaction was confirmed by TLC, and upon completion, the pH of the reaction product was adjusted to 1 with IN HC1 aqueous solution and extracted with Toyo.The organic layer was washed with water, dried over magnesium sulfate, and concentrated under reduced pressure to obtain the title target compound (1 g). Step 4 (d) Synthesis of N-cyclohexyl-2-(2-iodo-4-nitrophenyl)propenamide (N- cyclohexyl 1-2-(2-iodo-4-nitrophenyl)propenamide).

[0555] 2-(2-iodo-4-nitrophenyl)propanoic acid (2-(2-iodo-4-nitr opheny 1) pr opano ic acid, 1 g, 3 mmol), TBTU (1.4 g, 4.5 mmol, 1.5 eq) was added to 30 mL of DCM and stirred at room temperature for 50 minutes. TEA (0.83 ml, 6 mmol, 2 eq) and cyclohexanamine (41 ml, 3.6 mmol, 1.2 eq) were added to the reaction mixture and stirred at room temperature for 24 hours. Water was added to the reaction mixture and the mixture was extracted with distilled water. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the title target compound (1.4 g). Step 5 (e) 4- ((2- (1-(cyclohexylamino)- 1-oxopropan- 2 -yl)- 5-nitrophenyl)ethynyl)- N- ( 2- (piperidin- 1-yl)ethyl)benzamide (4- ( (2- (1-

[0556] Synthesis of (cyclohexylamino)— 1— oxopropan— 2— yl )-5-nitr ophenyl )ethynyl )— N— (2— (piper idin— 1-yl )ethyl )benzamide)

[0557] TEA (0.1 mL, 3 eq), Cui (1.4 mg, 3 mol%), and Pd[PPh3]4 (5.3 mg, 3 mol%) were added to a solution of N-cyclohexyl-2-(2-iodo-4-nitrophenyl)propenamide in MeCN (2 mL). The mixture was stirred at room temperature for 5 min under a nitrogen atmosphere. Then, 4-ethynyl-N-(2-(piperidin-1-yl)ethyl)benzamide (7 mg, 0.3 mmol, 1.0 eq) was added. The reaction mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The obtained residue was purified by column chromatography to obtain the target compound (0.15 g). Step 6 (f) 4- ( (5-amino-2- (1-(cyclohexylamino)-1-oxopropan-2-yl)phenyl)ethynyl)- N- (2- (piperidin-1-yl)ethyl)benzamide ( 4- ( ( 5-ami no-2- (1-

[0558] Synthesis of ( eye 1 ohexy 1 ami no)-l-oxopr opan-2-y 1 ) phenyl )ethynyl )— N— (2— (piper idin— 1— yl )ethyl )benzamide)

[0559] 4- ( (2- (1-(cyclohexylamino)- 1-oxopropan-2-yl)- 5-nitrophenyl)ethynyl)- N- (2-(piperidin- 1-yl)ethyl)benzamide ( 4-( (2-( 1-( cyclohexyl ami no)-l- oxopropan-2-y 1 ) — 5— nitr opheny 1 )ethyny 1 ) — N— ( 2— ( pi per idin— 1— y 1 ) et hy 1 ) benz amide , 0.26 g, 0.49 mmol , 1.0 eq) , Fe (0.27 g, 10 eq) NH4C1 (0.24 g, 10 eq) , EtOH (0.1 M) , H2O (0.1 M) were added and 90 °C for 2 hours. The reaction mixture became a black suspension, and when the reaction was completed, the reaction mixture was washed with water and extracted with DCM. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure to obtain the target compound as a crude product. Step 7 (g) Synthesis of phenyl (4-(1-(cyclohexylamino)-1-oxopropan-2-yl)-3-((4-((2-(piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)carbamate

[0560] 4- ( (5-amino-2- (1-(cyclohexylamino)-1-oxopropan-2-yl)phenyl)ethynyl)- N- (2-(piperidin-1-yl)ethyl)benzamide ( 4-( (5-ami no-2-( 1-( cyclohexyl ami no)-l- oxopropan-2-yl )phenyl )ethynyl )— N— (2— (piper idin— 1—yl )ethyl ) benz am ide , 0.24 g, 0.48 mmol , 1.0 eq) and Pyr idine (0.087 ml , 2 eq) were dissolved in THF (10 mL), and phenyl chloroformate (0.091 ml , 1.5 eq) was added while cooling the reaction solution with ice. The mixture was stirred at room temperature for 2 hours. After the reaction was completed, water (0.2 mL) was added to the reaction mixture, and the reaction mixture was concentrated under reduced pressure. The resulting residue was diluted with water and diluted with Toyo. The organic layer was washed with water, dried over magnesium sulfate, and concentrated under reduced pressure. The resulting residue was purified by column chromatography (silica gel, 0-5% MeOH / DCM) to obtain the target compound. Step 8 (h) Synthesis of 4-((2-(1-(cyclohexylamino)-1-oxopropan-2-yl)-5-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)ethynyl)-N-(2-(piperidin-1-yl)ethyl)benzamide (4— ((2— (1— ( ohexylamino ) — 1— oxopr opan-2-yl ) — 5— ( 3— ( 2— ( pyr i di n— 3— yl )ethyl )ureido)phenyl )ethynyl )— N— (2— (piper idin— 1— yl )ethyl )benzamide) Phenyl (4-(1-(cyclohexylamino)-1-oxopropan-2-yl)-3- ((4-((2-

[0561] (piperidin-1-yl)ethyl)carbamoyl)phenyl)ethynyl)phenyl)carbamate (phenyl (4-(1-

[0562] ( eye 1 ohexy 1 ami no ) — 1— oxopr opan— 2— y 1 )-3-( (4-( (2-(pi per i di n-1- yl )ethyl )carbamoyl )phenyl )ethynyl )phenyl )carbamate , 0.08 g, 0.13 mmol , 1.0 eq), 3- (2-aminoethyl )pyridine (3- (2-Aminoethyl )pyridine, 0.031 g, 0.25 mmol , 2 eq) and TEA ( 0.054 ml, 3 eq) were dissolved in THF (2 mL) and heated at 80 °C for 24 h. The reaction mixture was concentrated under reduced pressure, and the obtained residue was purified by column chromatography (silica gel, 0-5% MeOH / DCM) to obtain the target compound. P NMR (400 MHz, MeOD) 5 8.47 (d, J= 1.4 Hz, 1H) , 8.42 (dd, J= 4.8, 1.2 Hz, 1H), 7.89 (d, J = 8.3 Hz, 2H) , 7.81 - 7.79 (m, 1H) , 7.69 - 7.67 (m, 3H), 7.50 (d, J = 7.8 Hz, 1H) , 7.42 (dd, J = 7.6, 4.9 Hz, 1H) , 7.36 - 7.34 (m, 1H), 7.28 - 7.27 (m, 1H) , 4.13 (q, J= 7.2 Hz, 1H) , 3.64 - 3.61 (m, 3H) ,

[0563] 3.50 (t, J = 7.0 Hz, 2H), 3.37 (s, 1H), 2.92 (t, J = 6.9 Hz, 2H), 2.76 - 2.71 (m, 6H), 1.86 - 1.83 (m, 1H), 1.71 - 1.65 (m, 10H), 1.51 (d, J = 7.1

[0564] Hz, 3H), 1.39 — 1.00 (m, 6H). Example 32: Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(((1-methyl- 1H-pyrazol- 3-yl)methyl)sulfonyl)phenyl)-3-(2-(pyridin- 3 -yl)ethyl)urea (Scheme 27) Reaction Scheme 27. (a) Na2S, DMF, room temperature (rt), 16 hours (16h); (b) 3- (chloromethyl)-

[0565] 3-(chloromethyl)-1-methyl- IH-pyrazole, K2CO3, DMF, room temperature (rt), 16 h (16 h) (c) mCPBA, DCM, room temperature (rt), 48 h (48 hrs); (d) PdC12[PPh3]2, Cui, TEA, ACN, room temperature (rt), 16 h (16 h) (e) Fe, AcOH, 80 °C 2 h (2 h); (f) phenyl chloroformate, pyridine, THF, room temperature (RT), 3 h (3 h); (g) 2-(pyridin-3- yl)ethan-1-amine, TEA, THF, Reflux, 24 h (24 h) Step 1 (a) Synthesis of 2-iodo-4-nitrobenzenethiol

[0566] 1-Fluoro-2-iodo-4-nitrobenzene (1-f luoro-2-iodo-4-nitrobenzene, 5 g, 0.019 mol, 1 eq) was dissolved in DMF (25 ml), sodium disulfide (1.6 g, 0.021 mol, 1.1 eq) was added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, 200 ml of water was added to the reaction mixture, the pH was adjusted to 5 with IN HC1, and extracted several times with DCM. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure, and the resulting residue was purified by column chromatography (silica gel, 25% EA / HX) to obtain the target compound (4 g, 0.014 mol, 73%). Step 2 (b) Synthesis of 3-(((2-iodo-4-nitrophenyl)thio)methyl)-1-methyl-1H-pyrazole (3-(((2~i odo-4-nitr opheny 1 ) thio ) me t hy 1 )-1-methyl-IH-pyrazole)

[0567] 2-Iodo-4-nitrobenzenethiol (4 g, 0.014 mol, 1 eq) was dissolved in DMF (40 ml), and K2C03 (3.9 g, 0.028 mol, 2 eq) was added and stirred. 3-(chloromethyl)-1-methyl-1H-pyrazole (1.85 g, 0.014 mol, 1 eq) was added to the reaction mixture and stirred for 16 hours. The reaction was confirmed by TLC, and when the reaction was completed, water was added to the reaction mixture, and the resulting precipitate was filtered to obtain the target compound (4.9 g, 0.013 mol, 93%). Step 3 (c) Synthesis of 3-(((2-iodo-4-nitrophenyl)sulfonyl)methyl)-1-methyl-1H-pyrazole (3-(((2- iodo-4-nitr opheny 1 )sulfonyl )methyl )-1-methyl-1H-pyr azole)

[0568] 3-(((2-iodo-4-nitrophenyl)thio)methyl)-1-methyl-1H-pyrazole (3-(((2- iodo— 4— ni trophenyl)thio)methyl )—1— methyl— IH—pyrazole, 4.5 g, 0.012 mmol , 1 eq) was dissolved in DCM (200 ml), mCPBA (meta-chlorophenylbenzoic acid, 8.3 g, 0.048 mol, 4 eq) was added, and the mixture was stirred for 2 days. The reaction was confirmed by TLC, and additional mCPBA was added or, when the reaction was complete, water was added to dilute, and the mixture was neutralized with saturated aqueous NaHCOs solution and extracted several times with DCM. The organic layer was dried over magnesium sulfate, concentrated under reduced pressure, and recrystallized (MeOH / MC) to obtain the target compound (3 g, 0.0074 mol, 62%). Step 4 (d) Synthesis of 3-(((2-((4-fluorophenyl)ethynyl)-4-nitrophenyl)sulfonyl)methyl)-]-methyl- 1H-pyrazole (3-( ( (2-( (4-fluorophenyl)ethynyl)-4-nitr opheny 1)sulfonyl)methyl)-1-methyl- 1H-pyrazole)

[0569] 3-(((2-iodo-4-nitrophenyl)sulfonyl)methyl)-1-methyl-1H-pyrazole (3-(((2- iodo-4-nitr opheny 1 ) su 1 f ony 1 ) me t hy 1 ) - 1-me t hy 1 - IH-py r azole, 3 g, 0.0074 mol , 1 eq) was added with MeCN (30 ml) and TEA (3 ml, 0.022 mol, 3 eq). PdCl2[PPh3]2 (155 mg, 3 mol%) and Cui (42 mg, 3 mol%) were added to the reaction mixture and stirred at room temperature under a nitrogen atmosphere for 5 minutes. Then, 4-fluorophenyl-acetylene (1 g, 0.0088 mol , 1.2 eq) was added. The reaction mixture was stirred at room temperature for 16 hours and concentrated under reduced pressure. The obtained residue was purified by column chromatography (silica gel, 4% MeOH / DCM) to obtain the target compound (2 g, 0.005 mmol, 68%). Step 5 (e) Synthesis of 3-((4-fluorophenyl)ethynyl)-4-(((1-methyl- 1H-pyrazol-3-yl)methyl)sulfonyl)aniline (3-((4-fluorophenyl)ethynyl)-4-(((1-methyl- 1H-pyrazol-3-yl)methyl)sulfonyl)aniline)

[0570] 3- ( ( (2-( (4-fluorophenyl)ethynyl)-4-nitrophenyl)sulfonyl)methyl)-1-methyl-1H-pyrazole (3— ( ( (2— ( (4— f luorophenyl )ethynyl ) -4-nitr opheny 1 ) su 1 f ony 1 )methyl )—1— methyl - IH-pyrazole , 2 g, 0.005 mol 1 , 1 eq) was added AcOH (30 ml ) and iron (2.8 g, 0.05 mol , 10 eq) and stirred at 85°C for 2 to 3 hours. The catalyst was removed by filtration through Celite, and the filtrate was concentrated under reduced pressure. The residue was basified with aqueous potassium carbonate solution, extracted with Toyo, and the organic layer was concentrated under reduced pressure to obtain the target compound as a crude product. Step 6 (f) Synthesis of phenyl (3-((4-fluorophenyl)ethynyl)-4-(((1-methyl-1H-pyrazol-3-yl)methyl)sulfonyl)phenyl)carbamate

[0571] 3-((4-fluorophenyl)ethynyl)-4-(((1-methyl-1H-pyrazol-3-yl)methyl)sulfonyl)aniline (3-((4-f luorophenyl)ethynyl)-4-(((1-methyl-1H-pyrazol-3-yl)methyl)sulfonyl)aniline, 1.3 g, 0.0035 mol, 1 eq) and pyridine (Pyr idine, 0.56 ml, 0.0053 mol, 2 eq) were dissolved in THF, and phenyl chloroformate (Phenyl chloroformate, 0.66 ml, 0.0053 mol, 1.5 eq) was added while cooling the reaction solution with ice, and the mixture was stirred at room temperature for 3 to It was stirred for 4 hours. After the reaction was completed, water (0.1 mL) was added to the reaction mixture, and the reaction mixture was concentrated under reduced pressure. Water was added to the obtained residue, and extraction was performed with Toyo. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The obtained residue was purified by column chromatography (silica gel, 4-6% MeOH / DCM) to obtain the target compound (0.8 g,

[0572] 0.0016 mol, 46% was obtained. Step 7 (g) Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(((1-methyl-1H-pyrazol-3-yl)methyl)sulfonyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-fluorophenyl)ethynyl)-4-(((1-methyl -1H-pyrazol-3-yl)methyl)sulfonyl)phenyl)carbamate (phenyl (3-((4-fluorophenyl)ethynyl)-4-(((1-methyl-1H-pyrazol-3-yl)methyl)sulfonyl)phenyl)carbamate (phenyl (3-((4-fluorophenyl)ethynyl)-4-(((1-methyl-1H-pyrazol-3-yl)methyl)sulfonyl)phenyl)carbamate) )-4-( ( (1- methyl— 1H— pyr azo 1—3— yl) methy 1) sulfonyl ) phenyl) carbamate, 0.1 g, 0.2 mmol , 1.0 eq), 3- (2- aminoethyl ) pyridine (3- (2- Aminoethyl ) pyridine, 2 eq) and TEA (3 eq) were dissolved in THF (4 mL) and heated at 80 °C for 24 h. The reaction mixture was concentrated under reduced pressure, and the obtained residue was purified by column chromatography (silica gel, 0-5% MeOH / DCM) to obtain the target compound. Middle NMR (400 MHz, MeOD) 6 8.48 (s, 1H), 8.42 (m, 1H), 7.90 (s, 1H), 7.81 (d, J = 7.7 Hz, 1H), 7.74 -7.71 (m, 3H), 7.47 - 7.38 (m, 3H), 7.22 (t, J = 8.1 Hz, 2H), 6.13 (s, 1H), 4.75 (s, 2H), 3.77 (s, 3H), 3.52 (t, J = 6.7 Hz, 2H), 2.93 (t, J = 6.5 Hz, 2H).

[0573] MS(ESI) m / e MH +518.4 Example 33: Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(((1-methyl-1H-pyrazol-3-yl)methyl)sulfonyl)phenyl)-3-(imidazo[1,2-a]pyridin-7-ylmethyl)urea Compound of Example 33 was prepared in substantially the same manner as in Example 32, except that 1-{imidazo[1,2-a]pyridin-7-yl Imethanamine dihydrochloride# was used instead of 3-(2-aminoethyl)pyridine in step 7) compared to the synthesis of compound of Example 32. Medium NMR (400 MHz, DMSO) 6 9.41 (s, 1H), 8.50 (d, J = 6.9 Hz, 1H), 8.17 (s, 1H), 8.00 (s, 1H), 7.89 (s, 1H), 7.75 (m, 2H), 7.67 (d, J = 8.8 Hz, 1H), 7.58 (s, 1H), 7.52 (s, 1H), 7.46 (d, J = 8.7 Hz, 1H), 7.41 (s, 1H), 7.35 (t, J = 8.5 Hz, 2H), 7.12 (m, 1H), 6.86 (d, J = 6.9 Hz, 1H), 6.03 (s, 1H), 4.72 (s, 2H), 4.36 (d, J = 4.8 Hz, 2H), 3.73 (s, 3H).

[0574] MS(ESI) m / e MH+ 543.4 Example 34: 3-Amino-N-(2-((4-fluoroethyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)benzamide (Synthesis of 3-Amino-N-(2-((4-fluoroethyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)benzamide) (Synthesis Scheme 28)

[0575] Scheme 28. Reactants and Conditions: (a) 3-((tert-Butoxycarbonyl)amino)benzoic acid, DIPEA, HATU, DCM, room temperature (rt), 24 hours (24h); (b) 4N HCl in dioxane, 1,4-Dioxane, room temperature (rt), 24 hours (24h) Step 1 (a) Synthesis of tert-Butyl (3-((2-((4-fluorophenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)carbamoyl)phenyl)carbamate

[0576] 1-(4-Amino-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (0.05 g, 0.134 mmol, 1 eq) was dissolved in DCM (0.5 ml), and then 3-((tert-butoxycarbonyl)amino)benzoic acid (3-((tert-butoxycarbonyl)amino)benzoic acid,

[0577] 0.032 g, 0.134 mmol, leq), DIPEA (0.07 ml, 3 eq), HATU (0.061 g, 1.2 eq) were added, and the mixture was stirred at room temperature for 24 hours. After concentrating the reaction mixture, it was purified by reverse-phase column chromatography to obtain the title compound (0.047 g, 59.5%). Step 2 (b) Synthesis of 3-amino-N-(2-((4-fluorophenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)benzamide (3—amino—N—(2—((4-fluorophenyl)ethynyl)—4—(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)benzamid)e tert-butyl (3-((2-((4-fluorophenyl)ethynyl)-4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)carbamoyl)phenyl)carbamate, 0.047 g, 0.079 mmol, leq) was dissolved in 1,4-dioxane (1,4-Dioxane, 0.9 ml, 0.2 M), then 4N HCl * 4N HCl in dioxane, 0.2 ml, 0.79 mmol, 10 eq) was added, and the mixture was stirred at room temperature for 24 hours. After concentrating the reaction mixture, it was purified by reverse-phase column chromatography to obtain the title compound (2.62 mg, 7%). 1H NMR (400 MHz, MeOD) δ 8.44 (d, J = 21.9 Hz, 2H), 7.82 — 7.74 (m,

[0578] 2H), 7.68 (d, J = 2.3 Hz, 1H), 7.54 - 7.46 (m, 2H), 7.44 - 7.37 (m, 1H),

[0579] 7.32 (dd, J = 8.8, 2.3 Hz, 1H), 7.29 - 7.19 (m, 3H), 7.11 (t, J = 8.7 Hz,

[0580] 2H), 6.94 — 6.88 (m, 1H), 3.49 (t, J = 7.0 Hz, 2H), 2.91 (t, J = 6.9 Hz, 2H). Example 35: 4-Fluoro- N- (2- ((4-fluorophenyl)ethynyl)- 4- (3- (2- (pyridin-

[0581] 3 -ethyl)ureido)phenyl)benzamide (4-ho11101*0-si-(2-((4-bu11101*01)1161171)61:1171171)-

[0582] Synthesis of 4-(3-(2-(pyridin-3-yl)ethyl)ureido)phenyl)benzamide) (Synthetic Scheme 29) Scheme 29. Reactants and conditions: (a) 4-fluorobenzoic acid, DIPEA, HATU, DCM, room temperature (rt), 24 h;

[0583] 1-(4-amino-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(4-amino-3-((4-f 1 uor ophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea, 0.05 g, 0.134 mmol, leq) was dissolved in DCM (0.5 ml), and 4-fluorobenzoic acid (4-f luorobenzoic acid, 0.019 g, 0.134 mmol, leq), DI PEA (0.07 ml, 3 eq), and HATU (0.061 g, 1.2 eq) were added and stirred at room temperature for 24 h. The reaction mixture was concentrated and purified by reverse phase column chromatography to obtain the title compound (0.047 g, 59.5%). P NMR (400 MHz, MeOD) 5 8.46 (d, J = 24.0 Hz, 2H) , 8.12 - 8.03 (m, 2H), 7.81 (d, J = 7.8 Hz, 1H) , 7.72 (d, J = 2.4 Hz, 1H) , 7.67 (d, J = 8.8 Hz, 1H), 7.55 - 7.46 (m, 2H) , 7.45 - 7.39 (m, 1H) , 7.36 (dd, J = 8.8, 2.5 Hz, 1H), 7.28 (t, J = 8.7 Hz, 2H) , 7.12 (t, J = 8.8 Hz, 2H), 3.51 (t, J = 7.0 Hz, 2H), 2.93 (t, J = 6.9 Hz, 2H). Example 36: N- (2- ((4-fluorophenyl)ethynyl)-4- (3- (2- (pyridin-3-yl)ethyl)ureido)phenyl)propionamide (N- (2- ( (4- fl uor ophenyl )ethynyl )-4- (3 -

[0584] Synthesis of (2-(pyridin-3-yl)ethyl)ureido)phenyl)propionamide (Synthetic Scheme 30)

[0585] Scheme 30. Reagents and conditions: (a) propionic acid, DIPEA, HATU, DCM, room temperature (rt), 24 hours (24h);

[0586] 1-(4-amino-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(4-amino-3-((4-f 1 uor ophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea, 0.05g, 0.134mmol, leq) was dissolved in DCM (0.5ml), then propionic acid (0.01ml, 0.134mmol, leq), DI PEA (0.07ml, 3eq), and HATU (0.061g, 1.2eq) were added and stirred at room temperature for 24h. The reaction mixture was concentrated and purified by column chromatography (silica gel, DCM / MeOH) to obtain the title compound (0.039 g, 67%). P NMR (400 MHz, DMSO) 5 9.30 (s, 1H) , 8.58 (s, 1H) , 8.47 (d, J = 1.7 Hz, 1H), 8.44 (dd, J = 4.7, 1.4 Hz, 1H) , 7.73 (d, J = 2.4 Hz, 1H) , 7.70 - 7.62 (m, 3H), 7.52 (d, J = 8.7 Hz, 1H) , 7.37 - 7.28 (m, 3H) , 7.22 (dd, J = 8.8, 2.4 Hz, 1H), 6.23 (t, J = 5.7 Hz, 1H) , 3.37 (dd, J = 13.0, 6.9 Hz, 2H) ,

[0587] 2.79 (t, J = 7.0 Hz, 2H), 2.42 - 2.35 (m, 2H), 1.11 (t, J = 7.5 Hz, 3H). Example 37: Synthesis of 1-(3-((4-fluorophenyl)ethynyl)-4-(2-oxo-2-(piperidin-1-yl)ethyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (Scheme 31) Scheme 31 (a) TBTU, piperidine, TEA, DCM, room temperature (rt), 24 h (24 h) (b) Pd[PPh3]4, Cui, TEA, 1-ethynyl-4-fluorobenzene, ACN, room temperature (rt), 1 h (l h); (c) Zn, NH4CI, 1,4-dioxane, water (H2O), room temperature (rt), 24 h (24 h); (d) phenyl chloroformate, pyridine, THF, room temperature (RT), 2 h (2 h); (e) 2-(pyridin-3-yl)ethan-1-amine )etharrl-amine), TEA, THF, reflux, 24 h (24 h) Step 1 (a): Synthesis of 2-(2-iodo-4-nitrophenyl)-1-(piperidin-1-yl)ethan-1-one

[0588] 2-(2-iodo-4-nitrophenyl)acetic acid (1 g, 3.26 mmol) was stirred at room temperature for 50 minutes with 20 ml of DCM and TBTU (1.57 g, 1.5 eq). TEA (0.9 ml, 2 eq) and piperidine (0.38 ml, 1.2 eq) were added and stirred for 24 hours. Water was added to the reaction mixture, extracted with ethyl acetate, and the organic layer was dried over magnesium sulfate and concentrated under reduced pressure to obtain the target compound (1.1 g, 90%). Step 2 (b): Synthesis of 2-(2-((4-fluorophenyl)ethynyl)-4-nitrophenyl)-1-(piperidin-1—yl)ethan-1—one (2—(2—((4—f luorophenyl)ethynyl)—4—nitrophenyl)—1—(piperidin—1-yl )ethan-1-one) Triethylamine (1.22 ml, 3 eq), Cui (17 mg, 3 mol%) and Pd[PPh3]4 (62 mg, 3 mol%) were added to 2-(2-iodo-4-nitrophenyl)-1-(piperidin-1-yl)ethan-1-one (2-(2-iodo-4-nitrophenyl)—1—(piperidin—l—yl)ethan—l—one, 1.1 g, 2.94 mmol) in ACN (30 mL) was added to the solution. The mixture was stirred at room temperature for 5 minutes under a nitrogen atmosphere. Then, 1-ethynyl-4-f luorobenzene ( l-ethynyl-4-f luorobenzene , 0.42 mg, 1.2 eq) was added. The reaction mixture was stirred at room temperature for 1 hour and concentrated under reduced pressure. The obtained residue was purified by column chromatography (30% EA / HX) to obtain the target compound (0.84 g, 78%).Step 3 (c): Synthesis of 2-(4-amino-2-((4-fluorophenyl)ethynyl)phenyl)-1-(piperidin-1-yl)ethan-1-one.

[0589] 2- (2- ((4-fluorophenyl)ethynyl)-4-nitrophenyl)-1-(piperidin-1-yl)ethan-1-one (2— (2— ((4— fluorophenyl )ethynyl )—4— nitrophenyl)— 1— (piperidin— l—yl)ethan— l— one, 0.84 g, 2.3 mmol , 1.0 eq), Zn (1.5 g, 10 eq), NH4CI (1.37 g, 10 eq), dioxane:water (H2O) (12 ml , 3: l (v: v) , 0.02 M) were added and stirred at room temperature for 24 h. After the reaction was completed, the mixture was concentrated under reduced pressure to remove dioxane, and the reaction mixture was washed with water and a saturated NaHCOs solution and then extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure to obtain the title compound as a crude product. Step 4 (d): Synthesis of phenyl (3-((4-fluorophenyl)ethynyl)-4-(2-oxo-2-(piperidin-1—yl)ethyl)phenyl)carbamate

[0590] 2-(4-amino-2-((4-fluorophenyl)ethynyl)phenyl)-1-(piperidin-1-yl)ethan-1-one (2—(4—amino— 2—((4— f luorophenyl )ethynyl )phenyl )—1—(piperidin—l—yl )ethan— 1— one , 1.0 eq) and pyridine (0.42 ml , 2 eq) were dissolved in tetrahydrofuran (10 mL). The reaction solution was cooled with ice, and phenyl chloroformate (0.44 ml , 1.5 eq) was added. The mixture was stirred at room temperature for 2 h. After the reaction was completed, water (0.2 mL) was added to the reaction mixture, and the reaction mixture was concentrated under reduced pressure. The obtained residue was diluted with water and ethyl acetate. The organic layer was washed with water, dried over magnesium sulfate, and concentrated under reduced pressure. The residue obtained was purified by column chromatography (silica gel, 0-5% methanol / dichloromethane) to obtain the target compound. Step 5 (e): 1-(3-((4-fluorophenyl)ethynyl)-4-(2-oxo-2-(piperidin-1-yl)ethyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-f luorophenyl)ethynyl)—4— (2— oxo— 2— (piper idin—l—yl)ethyl)phenyl)-3- (2-

[0591] Synthesis of phenyl (3-((4-fluorophenyl)ethynyl)-4-(2-oxo-2-(piperidin-1-yl)ethyl)phenyl)carbamate ( phenyl (3- ((4-f 1 uor opheny 1 ) et hyny 1)-4-(2-oxo-2-

[0592] A mixture of (piperazin-l-yl)ethyl)phenyl)carbamate (0.2 g, 0.44 mmol, 1.0 eq), 3-(2-aminoethyl)pyridine (0.1 g, 2 eq), and triethylamine (0.18 ml, 3 eq) in THF (8 mL) was heated at 80 °C for 24 h. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by column chromatography (silica gel, 0-5% methanol / dichloromethane) to obtain the target compound. P NMR (400 MHz, DMSO) 5 8.59 (s, 1H), 8.47 (d, J= 1.8 Hz, 1H), 8.44

[0593] (dd, J = 4.7, 1.5 Hz, 1H) , 7.71 (d, J = 2.2 Hz, 1H) , 7.68 (dt, J = 7.6, 1.7

[0594] Hz, 1H), 7.64 - 7.57 (m, 2H), 7.37 - 7.27 (m, 3H), 7.23 (dd, J = 8.4, 2.3

[0595] Hz, 1H), 7.13 (d, J = 8.4 Hz, 1H), 6.25 (t, J = 5.6 Hz, 1H), 3.80 (s, 2H),

[0596] 3.47 - 3.42 (m, 4H) , 3.37 (dd, J = 12.9, 6.8 Hz, 2H) , 2.79 (t, J = 7.0 Hz,

[0597] 2H), 1.54 — 1.52 (m, J = 5.1 Hz, 2H), 1.39 (s, 4H). Example 38: N-(Piperidin-4-yl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (Syn-(Cheji6xin(Wu11-4-71)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide) synthesis (Synthesis Scheme 32)

[0598] Scheme 32. Reactants and conditions: (a) HATU, DIPEA, DCM, 0 °C to room temperature (rt), overnight; (b) PdCl2(PPh3)2, Cui, TEA, ACN, 60 °C overnight; (c) TFA, DCM, room temperature (rt), overnight Step 1 (a): Synthesis of tert-butyl 4-(4-ethynylbenzamido)piperidine-1-carboxylate (tert-butyl 4-(4-匕1±71171匕6112江111구(10)1江砂614(五116-1-031’1)0久71가6)

[0599] 4-Ethynylbenzoic acid (0.5 g, 3.42 mmol, 1 eq) was dissolved in DCM (34.2 ml, 0.1 M), HATU (1.55 g, 4.1 mmol, 1.5 eq) was added, and the mixture was stirred at room temperature for 10 minutes. tert-Butyl 4-aminopiperidine-1-carboxylate (0.821 g, 4.1 mmol, 1.5 eq) and DI PEA (1.326 g, 10.26 mmol, 3 eq) were added to the reaction mixture, and the mixture was stirred at room temperature overnight. After the reaction was completed, the mixture was extracted with EA and water (H2O). The organic layer was dried over MgS04, filtered, concentrated and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.769 g, 68.4%). Step 2 (b) tert-butyl 4- (4- ((5- (3- (2- (pyridin- 3 -yl)ethyl)ureido)-2- (pyridin- 4 -yl)phenyl)ethynyl)benzamido)piperidine- 1-carboxylate ( tert- butyl 4-

[0600] Synthesis of (4- ((5- (3- (2- (pyr idin- 3- yl )ethyl )urei do )-2-( py ridi n-4- yl )phenyl )ethynyl )benzamido)piperidine—l— carboxylate)

[0601] 1- (3-iodo-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (!- (3— iodo— 4— (pyridin— 4— yl)phenyl)— 3— (2— (pyridin— 3— yl)ethyl)urea, 0.05 g, 0.11 mmol, 1 eq), tert-butyl 4- (4-ethynylbenzamido)piperidine-1-carboxylate (tert- butyl 4- ( 4-ethynyl 1 benzamido)piperidine-1-carboxylate, 0.054 g, 0.165 mmol, 1.5 eq), PdCl2(PPh3)2 (0.002 g, 0.0033 mmol, 0.03 eq), Cui (0.00063 g, 0.0033 mmol, 0.03 eq), and TEA (0.022 g, 0.22 mmol, 2 eq) were dissolved in ACN (1.1 ml, 0.1 M) and degassed with nitrogen gas (N2 gas). The reaction mixture was heated to 60 °C and stirred overnight. After completion of the reaction, the mixture was extracted with DCM and water (C0). The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.021 g, 29.6%).Step 3 (c) Synthesis of N-(piperidin-4-yl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide (N-(piperidin-4-yl)-4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide) tert-butyl 4-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamide) tert-butyl 4-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzamido)piperidine-1-carboxylate (tert-butyl 4-(4-((5-(3- (2-(pyridin-3-yl)ethyl)urei do)-2-( pyridin-4- yl)phenyl)ethynyl)benzamido)piperidine—l— carboxylate, 0.021 g, 0.032 mmol, leq) was dissolved in DCM (1 ml, 0.07 M), TFA (0.036 g, 0.32 mmol, 10 eq) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the mixture was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN). (0.008 g, 49.3%) P NMR (400 MHz, DMSO) 5 9.22 (s, 1H) , 8.66 (d, J = 5.6 Hz, 2H) , 8.45 (dd, J = 11.1, 6.4 Hz, 3H), 8.33 (s, 1H) , 7.95 (s, 1H) , 7.87 (d, J = 8.2 Hz, 2H), 7.67 (dd, J = 12.2, 6.7 Hz, 3H) , 7.46 (dd, J = 14.7, 8.4 Hz, 4H) , 7.38 — 7.30 (m, 1H), 6.76 (s, 1H) , 3.96 (s, 1H) , 3.44 — 3.34 (m, 5H) , 2.85 — 2.77 (m, 3H), 1.93-1.85 (m, 2H) , 1.68-1.58 (m, 2H) .Example 39: 1-(3-((4-(4-aminopiperidine-1-carbonyl)phenyl)ethynyl)-4-.

[0602] (Pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea(1-(3-((4-(4-aminopiper idine-1-carbonyl )phenyl )ethynyl )— 4— (pyridin— 4— yl )phenyl )-3- (2 -

[0603] Synthesis of (pyr idin-3-yl)ethyl)urea (Synthetic reaction scheme 33)

[0604] Scheme 33. Reagents and conditions: (a) HATU, DIPEA, DCM, 0 °C to room temperature (rt), overnight; (b) PdCl2(PPh3)2, Cui, TEA, ACN, 60 °C, overnight; (c) TFA, DCM, room temperature (rt), overnight Step 1 (a) Synthesis of tert-butyl (1-(4-ethynylbenzoyl)piperidin-4-yl)carbamate

[0605] 4-Ethynylbenzoic acid (0.5 g, 3.42 mmol, 1 eq) was dissolved in DCM (34.2 ml, 0.1 M), HATU (1.55 g, 4.1 mmol, 1.5 eq) was added, and the mixture was stirred at room temperature for 10 minutes. Tert-butyl piperidin-4-yl carbamate (0.821 g, 4.1 mmol, 1.5 eq) and DI PEA (1.326 g, 10.26 mmol, 3 eq) were added to the reaction solution and stirred at room temperature overnight. After completion of the reaction, the mixture was extracted with EA and water (Compound 0). The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.640 g, 57%). Step 2 (b) Tert-butyl (1- (4- ((5- (3- (2-(pyridin-3-yl)ethyl)ureido)-2- (pyridin-4-yl)phenyl)ethynyl)benzoyl)piperidine-4 - Synthesis of tert-butyl (1-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)benzoyl)piperidin-4-yl)carbamate)

[0606] 1-(3-iodo-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3— iodo— 4— (pyridin— 4— yl)phenyl)— 3— (2— (pyridin— 3— yl)ethyl)urea, 0.1 g, 0.23 mmol, 1 eq), tert-butyl (1-(4-ethynylbenzoyl)piperidin-4-yl)carbamate (tert- butyl (1-(4-ethynylbenzoyl)piperidin-4-yl)carbamate, 0.113 g, 0.345 mmol, 1.5 eq), PdCl2(PPh3)2 (0.005 g, 0.0069 mmol, 0.03 eq), Cui (0.0013 g, 0.0069 mmol, 0.03 eq), TEA (0.047 g, 0.46 mmol, 2 eq) were dissolved in ACN (2.3 ml, 0.1 M) and degassed with nitrogen gas. The reaction mixture was heated to 60 °C and stirred overnight. After completion of the reaction, the mixture was extracted with DCM and water (ratio 0). The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.075 g, 50.6%). Step 3 (c) 1-(3-((4-(4-aminopiperidine-1-carbonyl)phenyl)ethynyl)-4-

[0607] Synthesis of tert-butyl (1-(4-((5-(3-(2-(pyridin-3-yl)phenyl)ethynyl)urea)-2-(pyridin-4-yl)phenyl)ethynyl)benzoyl)piperidin-4-yl)carbamate (1-(3-((4-(4-aminopiper idine-1-carbonyl)phenyl)ethynyl)—4— (pyridin—4— yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea) (0.068 g, 0.1 mmol, leq) was dissolved in DCM (1 ml, 0.1 MH), TFA (0.114 g, 1 mmol, 10 eq) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the mixture was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain the title compound (0.0337 g, 61.9%). P NMR (400 MHz, DMSO) 5 8.94 (s, 1H), 8.68 (s, 2H), 8.48 (d, J = 13.5 Hz, 2H), 7.97 - 7.90 (m, 3H), 7.73 (d, J = 7.8 Hz, 1H) , 7.68 (d, J = 5.3 Hz, 2H), 7.50 - 7.44 (m, 4H) , 7.41 - 7.36 (m, 3H) , 6.46 (t, J = 5.6 Hz, 1H), 4.57-4.27 (m, 1H) , 3.58 (s, 2H), 3.39 (dd, J = 12.7, 6.7 Hz, 3H), 3.13 (s, 1H), 2.81 (t, J = 6.9 Hz, 2H), 2.02-1.81 (m, 2H), 1.43 (s, 2H); MS MH +

[0608] 545.13 Example 40: Synthesis of N-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)phenyl)acetamide (N-(4-((5-(3-(2-(pyridin-3-yl)ethyl)ureido)-2-(pyridin-4-yl)phenyl)ethynyl)phenyl)acetamide) (Synthetic Reaction Scheme 34)

[0609] Scheme 34. Reagents and conditions: (a) Acetic anhydride, DCM, room temperature (rt), overnight; (b) PdC12(PPh3)2, Cui, TEA, ACN, 60 °C, overnight. Step 1 (a) Synthesis of N-(4-ethynylphenyl)acetamide (ci-(4-ethynylphenyl)acetamide)

[0610] 4—Ethynylaniline (4—ethynylani 1 ine, 0.5 g, 4.26 mmol, 1 eq) was dissolved in DCM (12.78 ml, 0.3 M), acetic anhydride (acetic anhydride, 0.477 g, 4.68 mmol, 1.1 eq) was added, and the mixture was stirred at room temperature. After completion of the reaction, the mixture was extracted with DCM and water (0). The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, EA / HEX) to obtain the title compound. Step 2 (b) Synthesis of N- (4-( (5- (3- (2- (pyridin- 3 -yl)ethyl)ureido)-2- (pyridin- 4-yl)phenyl)ethynyl)phenyl)acetamide

[0611] 1-(3-iodo-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3— iodo— 4— (pyridin— 4— yl)phenyl)— 3— (2— (pyridin— 3— yl)ethyl)urea, 0.1 g, 0.23 mmol , 1 eq), N-(4-ethynylphenyl)acetamide (N-(4-ethynylphenyl)acetamide, 0.055 g, 0.345 mmol , 1.5 eq), PdC12(PPhs)2 (0.005 g, 0.0069 mmol , 0.03 eq), Cui (0.0013 g, 0.0069 mmol , 0.03 eq), TEA (0.047 g, 0.46 mmol, 2 eq) was dissolved in ACN (2.3 ml, 0.1 L) and degassed with nitrogen gas (N2 gas). The reaction mixture was heated to 60 °C and stirred overnight. After completion of the reaction, it was extracted with DCM and water (Ratio 0). The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.047 mg, 43%). P NMR (400 MHz, DMSO) 5 10.12 (s, 1H) , 8.81 (s, 1H) , 8.66 (d, J = 6.0 Hz, 2H), 8.50 - 8.41 (m, 2H) , 7.85 (s, 1H) , 7.64 (ddd, J = 14.3, 13.3, 8.2 Hz, 5H), 7.42 (s, 2H) , 7.37 - 7.31 (m, 3H) , 6.34 (t, J = 5.6 Hz, 1H) , 3.39 (dd, J = 12.8, 6.7 Hz, 2H) , 2.80 (t, J = 7.0 Hz, 2H), 2.06 (s, 3H); MS

[0612] MH +476.2 Example 41: Synthesis of 1-(4-(6-aminopyridin-3-yl)-3-((4-fluorophenyl)ethynyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(4-(6-aminopyridin— 3— y 1 )-3-((4-fluorophenyl)ethynyl 1 )phenyl )-3-(2-(pyridin-3— yl)ethyl)urea) (Synthetic Reaction Scheme 35)

[0613] Scheme 35. Reagents and conditions: (a) PdC12(dppf ), K2CO3, 1,4-dioxane, H2O, 90 °C 1 hour (lh).

[0614] 1-(3-((4-fluorophenyl)ethynyl)-4-iodophenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-f luorophenyDet hyny 1)-4-iodophenyl)-3-(2-(pyr i din-3-yl)ethyl)urea, 0.1 g, 0.20 mmol, 1 eq) was dissolved in 1,4-dioxane (1,4-dioxane): water (H20)(3:1(V:V), 2 ml, 0.1 MH], and (6-aminopyridin-3-yl)boronic acid ((6-aminopyridin-3-yl)boronic acid, 0.055 g, 0.4 mmol, 2 eq), PdC12(dppf) (0.029 g, 0.04 mmol, 0.2 eq), K2CO3 (0.061 g, 0.44 mmol, 2.2 eq) were added and degassed with nitrogen gas. The reaction solution was heated to 90 °C and stirred for 1 hour. After completion of the reaction, it was filtered through a celite filter and extracted with EA and water (H2O). The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.053 g, 58.8%). P NMR (400 MHz, MeOD) 5 8.47 (d, J = 1.5 Hz, 1H), 8.42 — 8.38 (m,

[0615] 1H), 8.16 (d, J = 2.1 Hz, 1H), 7.79 (d, J = 7.9 Hz, 1H), 7.74 (dd, J = 8.6, 2.3 Hz, 1H), 7.68 (d, J = 2.2 Hz, 1H), 7.42 - 7.34 (m, 4H), 7.28 (d, J=8.5 Hz, 1H), 7.08 (t, J=8.8 Hz, 2H), 6.67 (d, J=8.6 Hz, 1H), 3.50 (t, J=7.0 Hz, 2H), 2.91 (t, J=7.0 Hz, 2H);MS MH + 452.08 Example 42: Synthesis of 1-(3-((4-aminophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-aminophenyl)ethyny 1)-4-(pyri din-Aryl)phenyl)-3-(2-(pyri idin-3-yl)ethyl)urea) (Synthetic Reaction Scheme 36) Scheme 36. Reagents and conditions: (a) Dimethyl 1-diazo-2-oxopropylphosphate, K2CO3, methanol (MeOH), room temperature (rt), 3 h; (b) PdC12(PPh3)2, Cu, Et3N, ACN, 90 °C, 2 h; (c) Zn, NH4C1, 1,4-dioxane, H2O, room temperature (rt), overnight. Step 1 (a) Synthesis of 1-ethynyl-4-nitrobenzene

[0616] 4-Nitrobenzaldehyde (0.3 g, 1.99 mmol, 1 eq) was dissolved in methanol (MeOH) (8 ml, 0.25 MH), dimethyl 1-diazo-2-oxopropyl phosphate (0.458 g, 2.388 mmol, 1.2 eq), K2CO3 (0.55 g, 3.98 mmol, 2 eq) were added, and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, the mixture was concentrated, diluted with EA (ethyl acetate), and extracted with brine and water in that order. The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, EA / HEX) to obtain the title compound (0.25 g. 85 %) was obtained. Step 2 (b) Synthesis of 1-(3-((4-nitrophenyl)ethynyl)-4-(pyridineyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea

[0617] 1-(3-iodo-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea

[0618] (1—(3— iodo— 4— (pyr idin— 4— yl ) phenyl )—3— (2— (pyr idin— 3— yl ) ethyl )urea, 0.2 g, 0.45 mmol, 1 eq) was dissolved in acetonitrile (ACN, 4.5 ml, 0.1 MH], and 1-ethynyl- 4-nitrobenzene (1-ethynyl- 4-nitrobenzene, 0.099 g, 0.675 mmol, 1.5 eq), PdC12(PPh3)2 (0.009 g, 0.0135 mmol, 0.03 eq), Cui (0.002 g, 0.0135 mmol, 0.03 eq), Et3N (0.091 g, 0.9 mmol, 2 eq) were added and then nitrogen Degassing was performed with gas (due gas). The reaction solution was heated to 90 °C and stirred for 2 hours. After completion of the reaction, it was filtered through celite and extracted with EA, 0. The organic layer was dried over MgSO, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.109 g, 52%). Step 3 (c) 1-(3-((4-aminophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-

[0619] Synthesis of (pyridin-3-yl)ethyl)urea (1-(3-((4-aminophenyl)ethynyl)-4-(pyridineyl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea)

[0620] 1-(3-((4-nitrophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea (1-(3-((4-nitrophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(2-(pyridin-3-yl)ethyl)urea, 0.020 g, 0.043 mmol, 1 eq) was dissolved in 1,4-dioxane (1,4-dioxane): water (H2O) (3:l(v / v))(2 ml, 0.02 MH], and then Zn (0.028 g, 0.43 mmol, 10 eq), NH4CI (0.023 g, 0.43 mmol, 10 eq) was added and stirred at room temperature overnight. It was washed with methanol and filtered under reduced pressure. The filtrate was concentrated and extracted with DCM: MeOH (9: l(v / v)), H2O. The organic layer was dried over MgS04, filtered, concentrated, and purified by column chromatography (silica gel, MeOH / DCM) to obtain the title compound (0.013 g, 69%). P NMR (400 MHz, MeOD) 5 8.56 (s, 2H), 8.43 (d, J = 25.2 Hz, 2H), 7.78 (d, J = 7.8 Hz, 1H), 7.70 (d, J = 4.3 Hz, 2H), 7.67 (d, J = 2.0 Hz, 1H), 7.39 (dt, J = 16.2, 5.2 Hz, 3H), 7.06 (d, J = 8.4 Hz, 2H), 6.61 (d, J = 8.4 Hz, 2H), 3.49 (t, J = 7.0 Hz, 2H), 2.90 (t, J = 7.0 Hz, 2H). Example 43: 1-(3-((4-aminophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-

[0621] Synthesis of (imidazo[1,2-a]pyridin-7-ylmethyl)urea (1-(3-((4-aminophenyl)ethynyl)-4-(pyridin-4-yl)phenyl)-3-(imidazo[l,2— a]pyridin— 7— ylmethyl)urea)

[0622] (Synthetic reaction scheme 37) Scheme 37. Reagents and conditions: (a) 4-ethynylaniline, PdC12(PPh3)2, Cui, TEA, ACN, 60 °C, overnight

[0623] 1-(Imidazo[1,2-a]pyridin-7-ylmethyl)-3-(3-iodo-4-(pyridin-4-yl)phenyl)urea (1-(imidazo[l,2-a]pyr idin-7-ylmethyl)-3-(3-iodo-4-(pyr idin-4-yl)phenyl)urea, 0.1 g, 0.213 mmol, 1 eq), 4-ethynylaniline (4-ethynylani 1 ine, 0.029 g, 0.256 mmol, 1.2 eq), PdC12(PPhs)2 (0.004 g, 0.006 mmol, 0.03 eq), Cui (0.001 g, 0.006 mmol, 0.03 eq), TEA (0.043 g, 0.426 mmol, 2 eq) was dissolved in ACN (2 ml, 0.1 M), heated to 60 °C, and stirred overnight. After completion of the reaction, the mixture was extracted with EA and 0. The organic layer was dried over MgSO4, filtered, concentrated, and purified by column chromatography (silica gel, DCM / MeOH) to obtain the title compound 190 (0.004 g, 3.6%). P NMR (400 MHz, MeOD) 5 8.54 (s, 2H) , 8.36 (d, J = 6.1 Hz, 1H) , 7.76 (s, 1H), 7.69 (d, J = 5.0 Hz, 3H) , 7.50 (s, 1H) , 7.43 (d, J = 6.6) Hz, 2H), 7.36 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 8.2 Hz, 2H), 6.89 (d, J = 7.0 Hz, 1H), 6.57 (d, J = 8.2 Hz, 2H), 4.45 (s, 2H).

[0624] <Experimental Example 1> NAMPT enzyme inhibition assay The NAMPT enzyme activity inhibition ability (IC50) of the compound was evaluated using the NAMPT Inhibitor Screening Assay Kit (# 7176-1) from BPS bioscience. Recombinant NAMPT protein was added to each well of a black 96-well plate, and dilution buffer was added to blank wells instead of NAMPT protein. After adding a 5-fold concentrated compound solution, the plate was pre-incubated at room temperature (approximately 22.0°C) for 30 minutes before starting the reaction to allow the compound and protein to bind before the enzymatic reaction. At this time, the concentration of NAMPT protein was adjusted according to the enzyme activity of each protein lot within the final concentration range of 4-10 ng / pl, and the test compound was serially diluted at a ratio of 1:3 from 1000 nM to 1 nM, and n=2 was tested for about 7 concentrations. A 2-fold concentrated test buffer containing ATP (final concentration 20 pM), Nicot inamide (final concentration 20 pM), Phosphoribosyl pyrophosphate (PRPP, final concentration 40 pM), and Ethanol (final concentration 1.5%) was added, and the plate was incubated at 30.0°C for 2 hours, and the fluorescence of the generated reactant was measured with excitation 340 nm and emission 460 nM. It was determined. NAMPT enzyme activity (%) was calculated by subtracting the measurement value of the blank well without NAMPT protein from the measurement value of all wells and then taking the measurement value of the control well without compound as 100%. IC50 was calculated using GraphPad Prism 9 software, and the results are shown in Table 1 below. In Table 1 below, A, B, and In are as follows.

[0625] A: NAMPT IC50 <0.5 |』M (0.5 pM or less)

[0626] B: 0.5 pM <NAMPT IC50 <1 pM(0.5 pM초과 1 pM 이하) C: NAMPT IC50 >1 pMd pM exceeds)

[0627] [Table 1] As confirmed in the table above, the compounds of the present invention exhibit excellent inhibitory activity against NAMPT, and sufficient inhibitory activity is observed even at low concentrations. Therefore, the compounds of the present invention may have excellent preventive or therapeutic effects against NAMPT-related diseases.

[0628] <Experimental Example 2> Cellular NAD measurement assay

[0629] The reduction of intracellular NAD+ and NADH levels due to the inhibition of NAMPT enzyme activity by compounds in HCT-116 cells was evaluated using the NAD / NADH-Glo™ Assay (Promega, G9072) from Promega. Cells were cultured in DMEM (Gibco, 11995-065) containing 10% FBS. The cells were dispensed at a density of 2,500 cells per well in a 96-well plate, and HCT-116 cells were cultured for 24 hours, and then treated with drugs using serum-free DMEM. First, the compounds were dissolved in 100% DMSO (Dimethyl sulfoxide), and then the compounds were diluted in the culture medium so that the final DMSO concentration was 0.5% for drug treatment, and the final concentration of the compounds was 0.1 nM - 100 nM. Afterwards, it was cultured for an additional 24 hours at 37°C C025% condition. To perform NAD / NADH-Glo™ Assay, the culture medium of drug-treated cells was removed, and 50 pL of DPBS per well was treated and left at room temperature for 5 minutes. NAD / NADH-Glo™ Detection Reagent (Reconstituted Luciferin Detection Reagent 1 mL, Reductase 5 |』L, Reductase Substrate 5 |』L, NAD Cyc 1 ng Enzyme 5 |』L, NAD

[0630] After preparing the required amount of Cycling Substrate 25 pL), 50 pL was treated per well. For cell lysis, the cells were incubated for 2 minutes at 400 rpm on a plate shaker, then incubated for 30 minutes at room temperature and luminescence was measured. The NAD+ / NADH level was calculated by subtracting the measurement value of the blank well without cells from the measurement value of all wells, and then using the 0.5% DMSO treatment control as the standard for 100%. The IC50 was calculated using GraphPad Prism 9 software, and the results are shown in Table 2 below. In Table 2 below, A, B, and C are as follows.

[0631] A: NAD IC 50 <10 nM (less than 10 nM)

[0632] B: 10 nM < NAD IC50 < IpM pM (over 10 nM, 1 pM or less)

[0633] C: NAD IC50 >1 pM (greater than 1 pM)

[0634] [Table 2]

[0635] As confirmed in the table above, the compounds of the present invention significantly inhibited NAD production in the HCT-116 cell line and significantly suppressed the concentration of intracellular NAD even at low concentrations. Therefore, the compounds of the present invention inhibit NAMPT and can sufficiently lower the concentration of NAD in cells even at low concentrations, and thus can be excellent in the prevention or treatment of diseases (e.g., cancer) that can be treated through the inhibition of NAMPT.

[0636] <Experimental Example 3> Tumor cytotoxicity assay

[0637] The anticancer efficacy of the compound was evaluated using the MTT assay in two cell lines, HCT-116 (colon cancer) and NCI-N87 (gastric cancer). HCT-116 cells were cultured in DMEM (Gibco, 11995-065) containing 10% FBS, and NCI-N87 cells were cultured in RPMI Medium 1640 (Gibco, 22400-089) containing 10% FBS and 25 mM HEPES. HCT-116 cells were seeded at a density of 2,500 cells per well and NCI-N87 cells were seeded at a density of 10,000 cells per well in 96-well plates, and the cells were cultured for 24 hours, and then treated with drugs using medium without FBS. First, the compounds were prepared in 100% DMSO (Dimethyl sulfoxide) at a concentration of 0.02 pM - 200 pM, and then the compounds were diluted in the culture medium so that the final DMSO concentration was 0.5% for drug treatment, and the final concentration range of the compounds was 0.1 nM - 1000 nM. After that, the cells were cultured for an additional 72 hours at 37°C C025%. To check for cytotoxicity, MTT (Sigma-Aldrich, M2128) was treated for 2 or 4 hours, and formazan produced by reduction by the enzymes of the mitochondria of living cells was measured using a Spark® Multimode Microplate Reader TECAN instrument. Cell viability (%) was relatively determined by subtracting the reference 650 nm absorbance from the 570 nm absorbance (Optical density, OD), and taking the 0.5% DMSO treatment control as the standard of 100%. The results were plotted as a function of compound concentration using GraphPad Prism 9 software to calculate CCso (50% cytotoxic concentration) values, and the results are shown in Table 3 below. In Table 3 below, A, B, and C are as follows.

[0638] A: CC5o <30 nM (30 nM or less)

[0639] B: 30 nM < CC5o <100 nM (over 30 nM and ≤100 nM)

[0640] C: CC50 >1 pM (over 1 pM)

[0641] [Table 3] As confirmed in the table above, the compounds of the present invention exhibit excellent cytotoxicity against colon cancer and gastric cancer cell lines even at low concentrations. Therefore, the compounds of the present invention are known to have excellent preventive or therapeutic effects against various diseases, such as cancer, by inhibiting NAMPT.

[0642] <2. Preparation of drug-linker conjugates> In order to confirm whether the cytotoxic drug moiety of the present invention is applicable to ADC, drug-linker conjugates were prepared as follows using the cytotoxic drug moieties of Examples 3, 10, 22, and 43, as examples. Example 44: Synthesis of linker-payload 1

[0643] To a solution of compound 1 (42.6 mg, 57.7 nmol, 1.0 equiv) dissolved in DMF (300 uL) were added compound 2 (compound of Example 10; 30 mg, 57.7 nmol, 1.0 equiv) and H0Bt (9.36 mg, 69.2 nmol, 1.2 equiv), DIEA (14.9 mg, 115 nmol, 20.1 uL, 2.0 equiv). The mixture was stirred at 25°C for 1 h. LC-MS showed one peak with the desired mass (MS cal.: 1117.8, MS observed: [M+H] + = 1118.9). The mixture was purified by prep-HPLC (TFA conditions). LCMS (MS cal.: 1118.5 [M+H]+) and HPLC confirmed that linker-payload l (25 mg, 21.7 μmol, 37.6% yield, 97.2% purity) was obtained as a white solid.

[0644] LCMS: MS cal.: 1117.8, MS observed: [M+H] + = 1118.5 Example 45: Synthesis of Linker-Payload 2 Step 1. To a solution of compound 3 (90 mg, 237 nmol, 1.0 equiv) and compound 4 (105 mg, 237 nmol, 1.0 equiv) in DMF (0.9 mL) was added DIEA (61.9 μL, 355 nmol, 1.5 equiv). The mixture was stirred at 25 °C for 1 h. LC-MS showed that compound 3 was completely consumed and one major peak with the desired mass (MS cal.: 706.3, observed MS: [M+H] + = 707.2) was detected. The reaction was purified by preparative HPLC (CTFA conditions). LCMS and HPLC confirmed that compound 5 (135 mg, 178 nmol, 74.9% yield, 93.1% purity) was obtained as a colorless oil.

[0645] LCMS: MS cal.: 706.3, MS observed: [M+H] += 707.2 Step 2. DIEA (61.9 μL, 355 nmol, 2.0 equiv) was added to a solution of compound 5 (135 mg, 178 nmol, 1.0 equiv) and bis(4-nitrophenyl) carbonate (324 mg, 1.07 mmol, 6.0 equiv) in DMF (2 mL). The mixture was stirred at 25°C for 1 h. LC-MS confirmed the complete consumption of compound 5 and the detection of a major peak with the desired mass (MS calculated: 871.3, MS observed: [M+H] + = 872.1). The reaction mixture was added to 20 mL of isopropyl ether and centrifuged to obtain compound 6. LCMS and HPLC results showed that compound 6 (130 mg, 144 nmol, 80.8% yield, 96.4% purity) was obtained as a colorless oil.

[0646] LCMS: MS cal.: 871.3, MS observed: [M+H] + = 872.1 Step 3. To a solution of compound 2 (compound of Example 10; 22 mg, 24.2 nmol, 1.0 equiv) and compound 6 (65.6 mg, 72.5 nmol, 3.0 equiv) dissolved in DMF (0.2 mL) were added HOBt (3.92 mg, 29.0 nmol, 1.2 equiv) and DIEA (8.42 yL, 48.3 nmol, 2.0 equiv). The mixture was stirred at 25°C for 1 h. LC-MS showed that compound 2 was completely consumed and one major peak with the desired mass (MS cal.: 1251.5, observed MS: [M+H] + = 1252.2) was detected. The mixture was purified by prep-HPLC (TFA conditions). LC-MS and HPLC confirmed that linker-payload 2 (22 mg, 17.5 u mol, 72.4% yield, 99.6% purity) was obtained as a white solid.

[0647] LCMS: MS cal.: 1251.5, MS observed: [M+Na] + = 1274.2 Example 46: Synthesis of linker-payload 3

[0648] Step 1. SOC12 (316 mg, 2.66 mmol, 193 uL) was added to a solution of compound 7 (1.00 g, 1.66 mmol) in THF (15 mL). The mixture was stirred at 25°C for 4 h. The completion of the reaction was confirmed by LC-MS. The reaction mixture was concentrated under reduced pressure to obtain a mixture, after which MTBE was added and the mixture was stirred at 25°C for 20 min to obtain compound 8 (1.01 g, 86.4% yield, 89.0% purity) as a yellow solid.

[0649] LCMS: m / z = 620.3 (M+H) + Step 2. Compound 8 (268 mg, 433 μmol) was added to a solution of compound 9 (compound of Example 3; 100 mg, 216 μmol) in DMF (2 mL). The mixture was stirred at 85°C for 6 hours. The completion of the reaction was confirmed by LC-MS. The reaction mixture was concentrated under reduced pressure to obtain a mixture, which was then purified by prep-HPLC (column: CDOl-Phenomenex luna C18 150*25*10 μm; mobile phase: [water (TFA)- ACN]; gradient: 22%-52% B, for 10 min). Compound 10 (80.1 mg, 31.1% yield, 97.7% purity, TFA) was obtained as a yellow solid.

[0650] LCMS: m / z = 1046.3 (M+l) +Step 3. Et2NH (0.2 mL) was added to a solution of compound 10 (TFA, 80.0 mg, 68.9 μmol) in DMF (1 mL). The mixture was stirred at 25°C for 0.5 h. The reaction was confirmed to be complete by LC-MS. The reaction mixture was concentrated under reduced pressure to obtain a residue. MTBE was added, and the mixture was stirred at 25°C for 10 min to obtain compound 11 (TFA, 60.1 mg, 92.7% yield) as a yellow solid.

[0651] LCMS: m / z = 823.9 (M+H) + Step 4. DIEA (15.6 mg, 121 μmol, 21.1 μL) was added to a solution of compounds 11 (50.0 mg, 60.6 μmol) and 12 (58.6 mg, 84.9 μmol) in DMF (2.50 mL). The mixture was stirred at 25°C for 1 h. The completion of the reaction was confirmed by LC-MS. The reaction mixture was concentrated to obtain a mixture, and then MeCN / MTBE = 1 / 1 (total 2.00 mL) was added, stirred at 25°C, and the resulting solid was dried under vacuum. The residue was purified by prep-HPLC (Column: CD04- Welch Ultimate C18 150*25*7 μm; Mobile phase: [water (TFA)-ACN]: gradient: 15%-35% B for 15 min). Linker-payload 3 (28.8 mg, 96.6% purity) was obtained as a white solid.

[0652] LCMS: m / z = 699.4 (M+H) +피 NMR (400 MHz, DMSO- ⑦) 6 10.12 (s, 1H) , 9.61 (s, 1H) , 9.17 (d, J = 6.8 Hz, 2H), 8.84 (d, J = 7.2 Hz, 1H) , 8.43 (d, J = 6.8 Hz, 2H) , 8.30 (d, J = 1.6 Hz, 1H), 8.08-8.18 (m, 2H) , 8.01 (d, J = 2.0 Hz, 2H) , 7.85 (d, J = 8.4 Hz, 1H), 7.76 (s, 1H) , 7.69 (dd, J= 8.4, 2.8 Hz, 3H) , 7.59 (dd, J= 8.8,

[0653] 2.4 Hz, 1H), 7.47-7.54 (m, 4H) , 7.39-7.46 (m, 2H) , 7.26 (t, J= 8.8 Hz, 2H) , 7.00 (s, 2H), 5.98 (d, J= 4.8 Hz, 1H) , 5.70-5.84 (m, 2H) , 5.29-5.53 (m, 2H) , 4.53 (d, J = 5.6 Hz, 2H) , 4.30-4.41 (m, 1H) , 4.15-4.25 (m, 1H) , 3.56-3.61 (m, 6H), 3.47 (s, 24H) , 3.35 (t, J = 5.6 Hz, 4H) , 3.14 (d, J = 5.6 Hz, 2H) , 2.92-3.01 (m, 2H) , 2.44-2.46 (m, 1H) , 2.39 (d, J = 6.4 Hz, 1H) , 2.33 (d, J = 7.6 Hz, 2H), 1.91-1.98 (m, 1H) , 1.53-1.68 (m, 2H) , 1.34-1.47 (m, 2H) , 0.83

[0654] (dd, J = 12.4, 6.8 Hz, 6H) . 실시예 47: 링커-페이로드 4의 합성 To a solution of compound 13 (158 mg, 141 μmol) and compound 14 (compound of Example 22; 80.9 mg, 141 μmol) in pyridine (2 mL) and DMF (2 mL) were added HOBt (38.2 mg, 282 limol) and DIEA (54.7 mg, 425 μmol, 73.8 uL). The mixture was stirred at 25°C for 2 hours. The formation of the desired compound was confirmed by LC-MS. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was then purified by prep-HPLC (Column: CD06-Waters Xbridge C18 150*40*10 μm; Mobile phase: [water (TFA)-

[0655] ACN]; gradient: 4%-34% B, for 10 min). Linker-payload 4 (15.50 mg, 6.6% yield, 98.5% purity) was obtained as a yellow solid.

[0656] LCMS: m / z = 1553.7 (M+H) + P NMR (400 MHz, DMSO—o6) 5 10.01 (s, 1H) , 9.34 (s, 1H) , 8.91 (d, J =

[0657] 6.0 Hz, 2H), 8.86 (t, J = 5.2 Hz, 1H), 8.81 (s, 1H), 8.76 (d, J = 5.2 Hz,

[0658] 1H), 8.37 (d, J = 8.0 Hz, 1H), 8.18 (d, J = 6.4 Hz, 2H), 8.13 (d, J = 7.2

[0659] Hz, 1H), 7.97-8.06 (m, 2H) , 7.84-7.95 (m, 4H) , 7.56-7.66 (m, 5H) , 7.52 (dd,

[0660] J = 8.4, 2.0 Hz, 1H), 7.33 (d, J = 8.4 Hz, 2H) , 6.99 (s, 2H) , 6.89 (t, J = 5.2 Hz, 1H), 6.07 (br s, 1H) , 5.05 (s, 2H) , 4.37 (dd, J= 13.2, 7.6 Hz, 2H) , 4.17-4.26 (m, 2H) , 3.55-3.69 (m, 8H) , 3.42-3.55 (m, 32H) , 3.29-3.39 (m, 5H) , 3.14 (q, J = 5.6 Hz, 3H), 2.90-3.09 (m, 5H) , 2.42-2.48 (m, 1H) , 2.39 (t, J = 6.4 Hz, 1H), 2.30-2.36 (m, 2H) , 1.96 (dq, J= 13.2, 6.8 Hz, 1H) , 1.54-1.76

[0661] (m, 2H), 1.31-1.50 (m, 2H), 0.84 (dd, J = 12.0, 6.8 Hz, 6H). Example 48: Synthesis of linker-payload 5

[0662]

[0663] Linker-payload 5 Step 1. 4-ethynylaniline (4-ethynylani 1 ine, 778 mg, 6.65 mmol) in THF

[0664] (20 ml) was added DIEA (1.72 g, 13.3 mmol, 2.32 ml). The mixture was stirred at 0 °C, and triphosgene (493 mg, 1.66 eq) was added dropwise under nitrogen. The mixture was stirred at 25 °C for 1 hour. A solution of compound 3 (2 g, 3.32 mmol) dissolved in DMF (20 ml) was added dropwise to the mixture. The reaction mixture was stirred at 25 °C for 2.5 hours. The completion of the reaction was confirmed by LC-MS. Water was added dropwise to the reaction mixture, and the precipitated solid was filtered under reduced pressure and dried in vacuo. The solid was purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 15 (713 mg, 28.8% yield).

[0665] LCMS: m / z = 745.5 (M+H) + Step 2. Compound 16 (450 mg, 0.959 mmol), Cui (18.26 mg, 0.096 mmol), Pd(PPhs)4 (110 mg, 0.096 mmol), and triethylamine (291 mg, 2.87 mmol, 0.401 ml) were added to a solution of compound 15 (1.07 g, 1.438 mmol) in DMF (15 ml), and the mixture was degassed with nitrogen gas. The mixture was heated to 40 。in and stirred for 3 hours. The completion of the reaction was confirmed by LC-MS. The reaction mixture was used in the next reaction without purification.

[0666] LCMS: m / z = 543.86 (1 / 2 M+H)+ Step 3. Piperidine (119 mg, 1.406 mmol, 0.139 ml) was added to a solution of compound 17 (509 mg, 0.469 mmol) in DMF (7.5 ml). The mixture was stirred at room temperature for 1 h. The completion of the reaction was confirmed by LC-MS. The reaction mixture was concentrated in vacuo and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 18 (42 mg, 10.4% yield).

[0667] LCMS: m / z= 864.6 (M+H) + Step 4. Compound 19 (40.23 mg, 0.058 mmol) and DIEA (12.57 mg, 0.097 mmol, 0.017 ml) were added to a solution of compound 18 (42 mg, 0.049 mmol) in DMF (1.8 ml). The mixture was stirred at room temperature for 1 h. The completion of the reaction was confirmed by LC-MS. The reaction mixture was purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain linker-payload 5 (containing the compound of Example 43; 1.7 mg, 2.4% yield).

[0668] LCMS: m / z= 1439.1 (M+H) +P NMR (400 MHz, DMSO) 6 10.02 (s, 3H) , 9.99 (s, 3H) , 8.65 (s, 6H) , 8.55 (s, 3H), 8.13 (s, 5H) , 7.94 (s, 5H) , 7.88 (s, 7H) , 7.62 (d, J= 8.9 Hz, 17H), 7.54 — 7.42 (m, 17H) , 7.34 (dd, J = 18.0, 8.2 Hz, 15H) , 6.95 (s, 7H) , 5.98 (s, 3H), 5.43 (s, 7H) , 5.09 (s, 6H) , 4.39 (s, 8H), 4.22 (d, J= 7.0 Hz, 4H), 3.59 (s, 11H), 3.47 (s, 105H), 3.17 (s, 8H), 2.98 (d, J= 28.0 Hz, 11H), 2.36 (d, J = 21.9 Hz, 8H), 1.93 (d, J = 17.1 Hz, 5H), 1.64 (d, J = 35.8 Hz, 8H), 1.37 (s, 8H), 0.84 (dd, J = 13.7, 6.1 Hz, 21H). Example 49: Synthesis of linker-payload 6

[0669] Step 1. Compound 21 (1.220 g, 9.91 mmol), HATU (2.260 g, 5.946 mmol), and DI PEA (1.280 g, 9.91 mmol, 1.72 ml) were added to a solution of compound 20 (2.768 g, 4.955 mmol) in THF (50 ml). The mixture was stirred at room temperature overnight. The completion of the reaction was confirmed by LC-MS. The reaction mixture was concentrated under reduced pressure, and DCMC (10 ml) was added, followed by hexane (200 ml). The precipitated solid was filtered under reduced pressure to obtain compound 22.

[0670] LCMS: m / z= 664.2 (M+H) +Step 2. DIPEA (3.466 g, 26.818 mmol, 4.671 ml) was added to a solution of compound 23 (1.131 g, 9.655 mmol) in THF (35 ml). Triphosgene (948 mg, 3.218 mmol) was added dropwise to the mixture at 0 °In. The mixture was stirred at 0 °In for 10 minutes. A solution of compound 22 (3.56 g, 5.364 mmol) in DMF (35 ml) was added dropwise to the mixture. The mixture was stirred at room temperature for 2 hours. The completion of the reaction was confirmed by LC-MS. The reaction mixture was concentrated under reduced pressure and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 24 (1.518 g, 35.1% yield).

[0671] LCMS: m / z= 807.4 (M+H) + Step 3. To a solution of compound 24 (1.676 g, 2.078 mmol) in DMF (14 ml) were added compound 16 (650 mg, 1.385 mmol), Cui (1.055 g, 5.540 mmol), DIPEA (358 mg,

[0672] After adding Pd(PPh3)4 (2.77 mmol, 0.483 ml) and Pd(PPh3)4 (1.120 g, 0.970 mmol), the mixture was degassed with nitrogen gas. The mixture was stirred at 40 。in for 1 hour. The completion of the reaction was confirmed by LC-MS. The reaction mixture was used in the next reaction without purification.

[0673] LCMS: m / z= 1148.02 (M+H) +Step 4. Piperidine (326 mg, 3.835 mmol, 0.379 ml) was added to a solution of compound 25 (1.468 g, 1.278 mmol) dissolved in DMF (20 ml). The mixture was stirred at 25 °C for 1 hour. It was confirmed by LC-MS that the reaction was complete. After concentrating the reaction mixture under reduced pressure, it was purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 26 (160 mg, 13.5% yield).

[0674] LCMS: m / z = 926.04 (M+H) + Step 5. Compound 27 (231.65 mg, 0.161 mmol), HATU (122 mg, 0.322 mmol), and DIPEA (31.2 mg, 0.241 mmol, 0.042 ml) were added to a solution of compound 26 (149 mg, 0.16 mmol) dissolved in DMF (2 ml). The mixture was stirred at 25 °C for 1 hour. It was confirmed by LC-MS that the reaction was complete. After concentrating the reaction mixture under reduced pressure, it was purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 28 (124 mg, 32.87% yield).

[0675] LCMS: m / z = 1173.90 (1 / 2 M+H)+ Step 6. Piperidine (22.1 mg, 0.26 mmol, 0.020 ml) was added to a solution of compound 28 (122 mg, 0.052 mmol) dissolved in DMF (6 ml). The mixture was stirred at 25 °C for 1 hour. It was confirmed by LC-MS that the reaction was complete. After concentrating the reaction mixture under reduced pressure, it was purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 29 (85 mg, 76.9% yield).

[0676] LCMS:m / z= 1062.71 (1 / 2 M+H)+ Step 7. Compound 30 (19.71 mg, 0.074 mmol) and DI PEA (10.34 mg, 0.080 mmol, 0.014 ml) were added to a solution of compound 29 (85 mg, 0.040 mmol) dissolved in DMF (1 ml). The mixture was stirred at 25 。in for 1 h. The completion of the reaction was confirmed by LC-MS. The reaction mixture was purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain linker-payload 6 (including the compound of Example 43; 40 mg, 43.9% yield).

[0677] LCMS: m / z= 1138.41 (1 / 2 M+H) + P NMR (400 MHz, DMSO) 5 10.00 (s, 1H) , 9.91 (s, 1H) , 9.02 (s, 1H) , 8.66 (s, 2H), 8.54 (s, 1H) , 8.43 (s, 1H) , 8.24 - 8.13 (m, 3H) , 8.04 (s, 1H) ,

[0678] 7.93 (s, 1H), 7.88 (s, 1H) , 7.85 (s, 1H) , 7.63 (d, J= 8.1 Hz, 4H) , 7.57 (s,

[0679] 1H), 7.50 (d, J = 8.2 Hz, 2H), 7.45 (t, J = 6.7 Hz, 3H), 7.38 (d, J = 8.3

[0680] Hz, 2H), 7.32 (d, J = 8.4 Hz, 2H) , 7.25 (d, J = 4.0 Hz, 4H) , 7.18 (d, J =

[0681] 4.3 Hz, 1H), 6.98 (s, 2H), 6.90 (d, J = 7.3 Hz, 2H), 5.09 (s, 2H), 4.52 (s,

[0682] 1H), 4.37 (d, J= 5.6 Hz, 2H), 4.16 (d, J= 6.3 Hz, 1H), 3.88 (dd, J= 16.4,

[0683] 8.9 Hz, 2H), 3.72 (dt, J= 17.5, 11.6 Hz, 4H) , 3.62 — 3.57 (m, 2H) , 3.50 (s,

[0684] 88H), 3.44 — 3.41 (m, 3H), 3.38 (t, J = 5.9 Hz, 2H), 3.23 (s, 3H), 3.17 (d, J = 5.3 Hz, 2H), 3.06 (d, J = 9.9 Hz, 1H), 2.86 — 2.77 (m, 1H), 2.40 (t, J = 7.4 Hz, 2H), 2.08 (d, J = 7.8 Hz, 2H), 1.92 — 1.80 (m, 1H), 1.71 (d, J = 7.8 Hz, 1H). Example 50: Synthesis of linker-payload 7

[0685]

[0686] Linker-payload 7 Step 1. Compound 31 (78.68 mg, 0.116 mmol), HATU (88.02 mg, 0.231 mmol), and DI PEA (22.44 mg, 0.174 mmol, 0.030 ml) were added to a solution of compound 18 (100 mg, 0.116 mmol) dissolved in DMF (2 ml). The mixture was stirred at 25 °C for 2 hours. The completion of the reaction was confirmed by LC-MS. The reaction mixture was concentrated under reduced pressure and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 32 (45 mg, 25.7% yield).

[0687] LCMS:m / z=755.20 (1 / 2 M+H)+ Step 2. Piperidine (7.61 mg, 0.089 mmol, 0.009 ml) was added to a solution of compound 32 (45 mg, 0.030 mmol) dissolved in DMF (1 ml). The mixture was stirred at 25 。in for 1 hour. The completion of the reaction was confirmed by LC-MS. The reaction mixture was concentrated under reduced pressure and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 33 (25 mg, 67.3% yield).

[0688] LCMS: m / z=1286.91 (M+H) + Step 3. CSI (9.12 mg, 0.064 mmol, 0.006 ml) was added to a solution of compound 34 (10 mg, 0.064 mmol) in DCM (6 ml). The mixture was stirred at 25 °C for 30 minutes. A solution of compound 33 (99.58 mg, 0.077 mmol) in DMF (0.24 ml) and TEA (32.61 mg, 0.322 mmol, 0.045 ml) were added to the mixture. The mixture was stirred at 25 °C for 30 minutes. The completion of the reaction was confirmed by LC-MS. The reaction mixture was purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain linker-payload 7 (including the compound of Example 43; 11 mg, 11.1% yield).

[0689] LCMS: m / z=774.15 (1 / 2 M+H) +P NMR (400 MHz, DMSO) 69.99 (d, J= 9.9 Hz, 2H) , 9.05 (s, 1H) , 8.67 (s, 2H), 8.11 (d, J= 7.7 Hz, 2H) , 7.86 (d, J= 8.8 Hz, 2H) , 7.68 — 7.59 (m, 6H), 7.48 (dd, J = 12.7, 6.4 Hz, 5H), 7.36 (d, J = 8.4 Hz, 2H), 7.31 (d, J = 8.9 Hz, 2H), 7.00 (d, J = 1.9 Hz, 3H), 5.97 (s, 1H) , 5.40 (s, 2H) , 5.09 (s, 2H), 4.41 (s, 2H), 4.23 (d, J = 7.2 Hz, 1H) , 4.03 (t, J = 6.4 Hz, 2H) , 3.59 (s, 2H), 3.51 — 3.47 (m, 33H) , 3.09 — 3.02 (m, 3H) , 2.67 (s, 1H) , 2.33 (s, 1H), 1.96 (s, 1H), 1.84 — 1.80 (m, 2H) , 1.74 — 1.66 (m, 1H) , 1.64 — 1.54 (m, 1H), 1.37 (s, 2H), 0.84 (dd, J = 13.1, 6.7 Hz, 6H). Example 51: Synthesis of linker-payload 8

[0690] Step 1. To a solution of compound 21 (7.28 g, 59.1 mmol) in CH2CI2 (60 mL) and MeOH (20 mL), EEDQ (14.6 g, 59.1 mmol) and compound 20 (5.5 g, 9.85 mmol) were added. The mixture was stirred at 25°C for 72 h. The completion of the reaction was confirmed by LC-MS. The reaction mixture was concentrated under reduced pressure. EtOAc (100 mL) was added to the concentrate and stirred at 25°C for 30 min to obtain compound 22 (6.1 g, 78% yield) as a yellow solid.

[0691] LCMS: m / z = 664.5 (M+H)+ Step 2. A solution of compound 22 (600 mg, 904 pmol) in THF (6 mL) was added with SOCl2

[0692] (537 mg, 4.52 mmol, 328 μL) at 0 °C. The mixture was stirred at 25 °C

[0693] for 1.5 h. It was confirmed by LCMS that the reaction was complete. The reaction mixture was recrystallized with MTBE (50 mL) and filtered under reduced pressure to obtain compound 35 as a yellow solid (580 mg, 94.1% yield).

[0694] LCMS: m / z = 682.1 (M+H) + Step 3. To a solution of compound 35 (561 mg, 823 pmol) and compound 36 (the compound of Example 3;

[0695] 190 mg, 411 pmol) in DMF (8 mL) were added KI (683 mg, 4.12 mmol), DIEA (133 mg, 1.03 mmol, 179 μL). The mixture was stirred at 25 °C for 6 h. It was confirmed by LC-MS that the reaction was complete. The reaction mixture was purified by prep-HPLC to obtain the mixed target product (TFA condition; column: F-Welch Xtimate C18 40*200mm 7μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 18%-58% B for 20 min). The mixture was purified by SFC (column: Daicel Chiralpak I BN 250mm*30mm*10μm; mobile phase: [CO2

[0696] EtOH: ACN = 3:1 (0.1% NH3-H2O)]; B%: 60%, isocratic elution mode) to obtain compound 37 as a yellow solid (85.0 mg, 53% yield).

[0697] LCMS:m / z = 1107.3 Step 4. Et2NH (355 mg, 4.85 mmol, 0.5 mL) was added to a solution of compound 37 (75 mg, 67.6 pmol) dissolved in DMF (3 mL). The mixture was stirred at 25 °C for 1 h. The reaction was confirmed to be complete by LC-MS. The reaction mixture was purified by prep-HPLC (column: F-Welch Xtimate C18 40*200mm 7um; mobile phase: [H20(0.1%TFA)-ACN]; gradient: 2%- 42% B for 20 min) to obtain compound 38 (65 mg, 96.1% yield, 100% purity) as a yellow solid.

[0698] LCMS: m / z = 885 (M) + Step 5. DIEA (18.9 mg, 146 pmol, 25.5 pL) and compound 39 (63.5 mg, 73.3 pmol) were added to a solution of compound 38 (65 mg, 73.3 pmol) in DMF (1 mL). The mixture was stirred at 25 。in for 1 hour. The completion of the reaction was confirmed by LC-MS. The reaction mixture was purified by prep-HPLC (column: F-Welch Xtimate C18 40*200mm 7um; mobile phase: [H20(0.1%TFA)-ACN]: gradient: 2%-42% B for 20 min) to obtain linker-payload 8 (36.16 mg, 30% yield, 100% purity) as a yellow liquid.

[0699] LCMS: m / z = 818.7 (M / 2) + Blood NMR (400 MHz, DMSO) 9.97 (s, 1H), 9.58 (s, 1H), 9.18 (d, J= 6.8

[0700] Hz, 2H), 8.84 (d, J = 6.4 Hz, 1H), 8.40-8.47 (m, 3H), 8.30 (d, J = 2.0 Hz, 1H), 8.15-8.20 (m, 2H), 8.12 (d, J = 1.6 Hz, 1H), 7.99-8.06 (m, 3H), 7.76 (s, 1H), 7.67-7.72 (m, 3H), 7.59 (dd, J = 8.8, 2.0 Hz, 1H), 7.49-7.55 (m, 4H), 7.44 (d, J = 7.2 Hz, 1H) , 7.35 (t, J = 6.0 Hz, 1H) , 7.27-7.30 (m, 1H) , 7.24-7.27 (m, 5H) , 7.16-7.21 (m, 1H) , 7.00 (s, 2H) , 5.79 (s, 2H), 4.53 (d, J = 5.6 Hz, 2H), 4.47-4.50 (m, 1H) , 3.88 (dd, J = 12.4, 6.0 Hz, 2H) , 3.74- 3.83 (m, 2H), 3.68 (d, J = 5.6 Hz, 3H) , 3.55-3.64 (m, 10H) , 3.44-3.48 (m, 34H), 3.35 (t, J = 6.0 Hz, 4H) , 3.12-3.16 (m, 2H) , 3.05 (dd, J = 13.6, 4.0 Hz, 1H), 2.79-2.85 (m, 1H) , 2.64-2.69 (m, 1H) , 2.38 (t, J = 6.4 Hz, 2H),

[0701] 2.30-2.34 (m, 2H) Example 52: Synthesis of linker-payload 9 Step 1. Compound 40 (2.00 g, 4.39 mmol) and compound 41 (2.73 g, 8.78 mmol) were dissolved in DCM (20 ml) and MeOH (8 ml), then EEDQ (2.17 g, 8.78 mmol) was added and stirred at 25 °C for 3 hours. After completion of the reaction, the mixture was concentrated and purified by column chromatography.

[0702] (silica gel, petroleum ether / EA) to obtain compound 42 (2.43 g, 69.84% yield) as a white solid.

[0703] LCMS: m / z = 749.1 (M+H) + Step 2. Compound 42 (1.00 g, 1.34 mmol)> Dissolve in DCM (10 mL), add SOC12 (794 mg, 6.68 mmol), and stir at 0 。In for 2 hours. After confirming that the reaction was complete by LC-MS, the reaction mixture was concentrated to obtain compound 43 (lg, crude) in the form of a colorless oil.

[0704] LCMS:m / z = 767.3(M+H)+ Step 3. Compound 43 (997 mg, 1.30 mmol) and compound 44 (compound of Example 3; 200 mg, 433 pmol) were dissolved in DMF (10 mL), KI (431 mg, 2.60 mmol) and DIEA (140 mg, 1.08 mmol) were added, and the mixture was stirred at 25 。in for 4 hours. After confirming that the reaction was complete by LC-MS, the reaction mixture was concentrated and purified by prep-HPLC (TFA conditions) to obtain compound 45 (80.0 mg, 5.02% yield) as a yellow solid.

[0705] LCMS: m / z = 1192 P NMR: (400 MHz, DMSO—⑦) <59.54 (s, 1H), 9.17 (d, J = 6.8 Hz, 2H), 8.88 (s, 1H), 8.83 (d, J = 7.2 Hz, 1H), 8.43 (d, J = 6.8 Hz, 2H), 8.29 (d, J = 1.6 Hz, 1H), 8.11 (s, 2H), 8.00 (d, J = 2.0 Hz, 1H), 7.87 (d, J = 7.2 Hz, 2H), 7.75 (s, 1H), 7.63-7.71 (m, 3H) , 7.49-7.60 (m, 3H), 7.36-7.45 (m, 4H), 7.23-7.34 (m, 6H), 7.18 (d, J = 8.4 Hz, 1H), 5.78 (s, 2H), 5.64 (d, J = 8.0 Hz, 1H), 5.49 (t, J = 9.6 Hz, 1H) , 5.17-5.22 (m, 1H) , 5.06 (t, J = 9.6 Hz, 1H), 4.74 (d, J = 10.0 Hz, 1H) , 4.53 (br d, 7 = 5.6 Hz, 2H) , 4.25-4.32 (m, 2H), 4.16-4.24 (m, 1H) , 3.60 (s, 3H) , 3.24-3.29 (m, 4H) , 1.97-2.03 (m, 9H) Step 4. Compound 45 (70.0 mg, 58.6 pmol) was dissolved in THF (2.25 mL) and H2O

[0706] After dissolving in (0.75 mL), Li0H-H20 (7.39 mg, 176 pmol) was added and stirred at 25。in for 6 hours. After confirming that the reaction was complete by LC-MS, MeCN (40 mL) was added to the reaction mixture, and the resulting precipitate was filtered under reduced pressure to obtain compound 46 (35.0 mg, crude) as a brown solid.

[0707] LCMS: m / z = 830.2 Step 5. Compound 46 (35.0 mg, 42.1 pmol) and compound 47 (25.9 mg, 84.2 pmol) were dissolved in DMF (5 mL) and stirred at 25 。in for 1 h. After confirming the completion of the reaction by LC-MS, the reaction mixture was concentrated and purified by prep-HPLC (TFA conditions) to obtain linker-payload 9 (2.10 mg, 4.78% yield, 98.15% purity) as a yellow solid.

[0708] LCMS: m / z = 1023.5 P NMR: (400 MHz, DMSO—⑦) <59.47 (s, 1H), 9.15 (d, J = 6.8 Hz, 2H),

[0709] 9.12 (s, 1H), 8.73 (d, J = 7.6 Hz, 1H), 8.44 (d, J = 6.4 Hz, 2H), 8.36 (s,

[0710] 1H), 8.17 (s, 1H), 8.00 (s, 1H), 7.94 (s, 1H), 7.85 (t, J = 6.0 Hz, 1H),

[0711] 7.63-7.72 (m, 2H), 7.59 (d, J = 8.8 Hz, 1H), 7.51-7.57 (m, 2H), 7.26 (t, J = 8.8 Hz, 5H), 7.17 (d, J = 8.8 Hz, 1H), 6.98 (s, 2H) , 5.79 (s, 3H) , 5.24- 5.49 (m, 2H), 4.93 (d, J = 7.6 Hz, 1H) , 4.48 (d, J = 5.6 Hz, 2H) , 3.92 (d, J = 9.6 Hz, 1H), 2.67 (s, 2H) , 2.55 (s, 2H), 2.40-2.41 (m, 1H), 2.33 (d, J = 0.8 Hz, 2H), 2.01 (s, 2H), 1.42-1.48 (m, 4H), 1.23 (s, 2H), 1.13-1.18 (m, 2H) Example 53: Synthesis of linker-payload 10 Step 1. Compound 48 (1.00 g, 1.66 mmol) was dissolved in THF (15 mL), and then SOC12

[0712] (316 mg, 2.66 mmol, 193 pL) was added, and the mixture was stirred at 25 °C for 4 hours. After confirming the completion of the reaction by LC-MS, the reaction mixture was concentrated, MTBE (20 mL) was added, and the mixture was stirred for 20 minutes to obtain Compound 49 (1.06 g, 89.9% yield) in the form of a white solid.

[0713] LCMS m / z = 620.1 (M+H) + Step 2. Compound 49 (464 mg, 748 pmol) was dissolved in DMF (8 mL), and then KI (827 mg, 4.98 mmol), Compound 50 (the compound of Example 3; 230 mg, 498 pmol), and DIEA (129 mg, 997 pmol, 174 pL) were added, and the mixture was stirred at 25 °C for 16 hours. After confirming the completion of the reaction by LC-MS, the reaction mixture was purified by prep-HPLC (TFA conditions) and then by prep-SFC (CO2 / MeOH = 3:1 (0.1% FA:NH3 in MeOH)) to obtain Compound 51 (153 mg, containing 38.9% of Compound 52).

[0714] LCMS m / z = 1045.6 Step 3. Compound 51 (153 mg, 146 pmol) was dissolved in DMF (2 mL), and then piperidine (74.7 mg, 877 pmol, 86.7 pL) was added, and the mixture was stirred at 25 °C for 1 hour. After confirming the completion of the reaction by LC-MS, the reaction mixture was purified by prep-HPLC (TFA conditions) to obtain Compound 52 (112 mg, 90.8% yield) in the form of a yellow solid.

[0715] LCMS m / z = 823.2 Step 4. Compound 52 (102 mg, 124 pmol) was dissolved in DMF (5 mL), and then HATU (94.1 mg, 248 pmol), DIEA (32.0 mg, 248 pmol, 43.1 pL), and compound 53 (178 mg, 124 pmol) were added and stirred at 25°C for 1 hour. After confirming that the reaction was complete by LC-MS, the reaction mixture was used in the next reaction without purification.

[0716] LCMS m / z = 1122.7 (M / 2) Step 5. Piperidine (105 mg, 1.24 mmol, 122 pL) was added to the reaction mixture of Step 4 and stirred at 25°C for 1 h. After confirming the completion of the reaction by LC-MS, the reaction mixture was purified by prep-HPLC (TFA conditions) to obtain compound 55 (17.1 mg, 98.3% yield) as a yellow gum.

[0717] LCMS m / z = 1011.7 (M / 2) Step 6. Compound 55 (17.1 mg, 8.40 pmol) was dissolved in DMF (1.5 mL), and compound 56 (2.68 mg, 10.1 pmol) and DIEA (2.17 mg, 16.8 pmol, 2.93 pL) were added and stirred at 25°C for 1 h. After confirming the completion of the reaction by LC-MS, the reaction mixture was purified by prep-HPLC (TFA conditions) to obtain linker-payload 10 (10.8 mg, 56.2% yield, 100% purity) as a yellow gum.

[0718] LCMS m / z = 1087.6 (1 / 2M+H) +

[0719] HRMS m / z = 2173.1 Example 54: Synthesis of linker-payload 11

[0720] Step 1. Dissolve compound 58 (2.1 g, 0.018 mol) in THF (67 ml, 0.15 M).

[0721] DI PEA (8.7 ml, 0.05 mol) was added and triphosgene (1.78 g, 0.006 mol) was added at 0°C under nitrogen and stirred for 10 minutes at 0°C. Compound 57 (7.394 g, 0.01 mol) was added to anhydrous

[0722] Dissolve in DMF (67 ml, 0.15 M) and slowly dropwise add to the reaction mixture and stir at 25 。in for 1 hour. Water was added to the reaction mixture, and the precipitated solid was filtered under reduced pressure and dried in vacuo. The solid was purified by column chromatography (silica gel, HX / EA) to obtain compound 59 (5.995 g, yield 67.2%) as a white solid*. P NMR (400 MHz, DMSO) 6 9.99 s, 1H), 8.81 (s, 1H), 7.93 s, 1H),

[0723] 7.88 (d, J= 7.5 Hz, 2H), 7.69 (d, J = 7.4 Hz, 2H), 7.47 (d, J= 8.4 Hz, 2H),

[0724] 7.40 (t, J= 7.6 Hz, 5H), 7.31 (t, 7 = 6.7 Hz, 2H), 7.20 (d, J= 8.4 Hz, 1H),

[0725] 7.09 (d, J= 8.4 Hz, 1H), 5.60 (d, J = 7.8 Hz, 1H), 5.50 (t, J= 9.7 Hz, 1H),

[0726] 5.18 (t, J = 8.8 Hz, 1H), 5.10 — 5.03 (m, 3H), 4.73 (d, J = 10.0 Hz, 1H),

[0727] 4.32 — 4.27 (m, 2H), 4.24 — 4.18 (m, 1H) , 4.06 (s, 1H) , 3.63 (s, 3H) , 3.33 — 3.25 (m, 2H), 2.00 (d, J = 4.5 Hz, 9H) . Step 2. Compound 59 (1.996 g, 2.237 mmol), compound 60 (0.7 g, 1.492 mmol), Cui (85.22 mg, 0.447 mmol), Pd(PPh3)4 (314 mg, 0.447 mmol)> Dissolve in anhydrous DMF (75 ml, 0.02 M), then add TEA (2 ml, 14.916 mmol)> and degas with nitrogen gas. The mixture was heated to 40 。in and stirred for 2 hours. The reaction was confirmed to be complete by LC-MS, and the reaction mixture was concentrated and purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 61 (0.681 g, 45.1% yield) as a white solid. P NMR (400 MHz, DMSO) 6 10.01 (s, 1H), 9.36 (s, 1H), 7.92 (s, 2H),

[0728] 7.54 — 7.41 (m, 5H) , 7.34 (d, J = 7.4 Hz, 3H) , 7.25 (d, J = 8.5 Hz, 2H) , 7.13 (d, J= 8.2 Hz, 1H), 6.89 (d, J= 5.6 Hz, 1H) , 5.64 (d, J= 7.3 Hz, 1H) , 5.51 (t, J = 9.5 Hz, 1H), 5.19 (t, J = 8.2 Hz, 1H) , 5.14 — 5.02 (m, 3H) ,

[0729] 4.75 (d, J = 9.9 Hz, 1H) , 4.33 (s, 2H) , 3.64 (s, 4H) , 2.55 (s, 1H) , 2.14 —

[0730] 2.08 (m, 1H), 2.02 (d, J = 3.5 Hz, 9H). Step 3. Compound 61 (0.681 g, 0.674 mmol) was dissolved in THF: H2O (v / v=3:1, 34 ml, 0.02 M), then Li0H-H2O (0.17 g, 4.04 mmol) was added and stirred at 25 。in for 1 h. The completion of the reaction was confirmed by LC-MS, and MeCN (400 ml) was added to the reaction mixture. The precipitated solid was filtered under reduced pressure and dried in vacuo. The solid was purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN) to obtain compound 62 (0.279 g, 47.5% yield).

[0731] LCMS: m / z = 871 (M+H) + blood NMR (400 MHz, DMSO) 6 10.02 (s, 1H) , 9.56 (s, 1H) , 9.26 (s, 1H) , 8.67 (d, J= 4.1 Hz, 2H), 8.51 (d, J= 6.8 Hz, 1H) , 8.19 (d, J = 5.5 Hz, 3H) , 7.90 (s, 2H), 7.66 (d, J = 5.2 Hz, 2H) , 7.58 — 7.39 (m, 6H) , 7.33 (d, J = 8.5 Hz, 2H), 7.17 (s, 3H) , 6.87 (d, J = 6.8 Hz, 1H), 5.10 (s, 2H), 4.70 (d, J = 6.9 Hz, 1H), 4.36 (d, J = 4.9 Hz, 3H), 3.10 (s, 3H), 2.55 (s, 2H). Step 4. Dissolve compound 62 (0.126 mg, 0.145 mmol) in DMF (1.45 ml, 0.1M), add compound 63 (53.52 mg, 0.174 mmol), DI PEA (0.076 ml, 0.434 mmol), and stir at 25 。in for 1 hour. The reaction was confirmed to be complete by LC-MS, purified by reverse-phase column chromatography (0.1% formic acid in H2O / ACN), and lyophilized to obtain linker-payload 11 (compound of Example 43; 21 mg, 13.6% yield).

[0732] LCMS: m / z=1064.6 (M+H) +P NMR (400 MHz, DMSO) 6 9.99 (s, 1H) , 9.08 (s, 1H) , 9.00 (s, 1H) , 8.66 (d, J = 5.8 Hz, 2H) , 8.50 (d, J = 7.0 Hz, 1H) , 8.23 ​​(s, 1H) , 7.93 — 7.84 (m, 3H), 7.64 (d, J = 5.7 Hz, 2H) , 7.53 (s, 1H) , 7.51 — 7.39 (m, 5H) , 7.32 (d, J= 8.5 Hz, 2H) , 7.10 (s, 2H) , 6.99 (s, 2H) , 6.90 (s, 1H) , 6.87 (d, J = 7.0 Hz, 1H), 5.84 (d, J = 3.7 Hz, 1H) , 5.30 (d, J = 4.8 Hz, 1H) , 5.07 (s, 2H), 4.86 (d, J = 6.8 Hz, 1H) , 4.36 (d, J = 6.0 Hz, 2H) , 3.87 (d, J = 9.8 Hz, 1H), 2.55 (d, J = 7.4 Hz, 2H) , 2.02 (t, J = 7.3 Hz, 2H) , 1.45 (dd, = 14.3, 7.3 Hz, 4H) , 1.21 — 1.11 (m, 2H) .

[0733] <3. Preparation of antibody-drug conjugate> Example 55: Conjugation method

[0734] (1) Trastuzumab-linker-payload conjugation experimental procedure) Trastuzumab-linker-payload conjugation was performed as follows. Trastuzumab dissolved in initial buffer (4.23 mM histidine / histidine-HC1, 55.77 mM trehalose dihydrate, 0.0085% polysorbate 20) was pipetted into a 50 mL tube. ) Reduction buffer was added to make the mAb concentration 8.89-9.0 mg / mL. ) TCEP was added for reduction. The reaction vial was placed in an incubator-shaker at 22 °C and reacted at 60 rpm for 18 h or at 37 °C, 300 rpm for 2-3 h. ) After reduction, DMA solvent was added to the sample to make the organic solvent ratio 10% (v / v). ) Linker-payload dissolved in DMA was added to the final mAb concentration of 8 mg / mL. The mixture was reacted at 22-25 °C for 1 hour. After 1 hour, DAR was confirmed by LC-MS, and the sample was purified by Zeba Spin desalting column (40K, 10mL*3) or HighTrap desalting column. ) Next, the sample was concentrated and buffer exchanged with 20 mM histidine / histidine HC1, pH 5.5 using Ami con (50 KDa, 15 mL), and then SEC purity and free drug concentration were confirmed.

[0735] (2) ADC purification using Zeba Spin desalting column

[0736] The Zeba Spin desalting column was used after pretreatment as follows.

[0737] 1) After removing the bottom stopper of the column, centrifugation (700 g, 2 minutes) was performed to remove the storage solution.

[0738] 2) The column was sterilized by applying 5 mL of 0.5 M NaOH onto the resin and left for 30 minutes.

[0739] 3) After centrifugation (700 g, 2 minutes), the eluate was discarded.

[0740] 4) After applying 5 ml of formulation buffer to the resin, centrifugation (700 g, 2 min) was performed, and the eluate was discarded. This step was repeated two more times until the pH of the eluate became the same as that of the formulation buffer. Finally, centrifugation was performed for 5 minutes.

[0741] 5) The column was transferred to a new collection tube and the antibody-drug conjugate mixture was applied onto the column resin.

[0742] 6) The antibody-drug conjugate was obtained by centrifugation (700 g, 4 minutes).

[0743] (3) Purification of ADC using Amicon® Ultra Centrifugal Filter Unit Low molecular weight substances were removed using an ultra-low molecular weight filtration membrane. It is recommended to use an ultra-low molecular weight filtration membrane with a molecular weight of 50 kDa. The Amicon Ultra Centrifugal Filter Unit was used in the following manner.

[0744] 1) Wash the centrifugal filter unit with distilled water.

[0745] 2) Equilibrated with 20 mM histidine buffer (pH 5.5).

[0746] 3) Up to 12 mL of the antibody-drug conjugate mixture was added to an Ami con® Ultra filter device and centrifuged to concentrate to approximately 3 mL. 9 mL of 20 mM histidine buffer (pH 5.5) was added and buffer exchange was performed until the amount of water discarded was 63 mL to remove low-molecular-weight substances. The remaining solution was concentrated to obtain approximately 9 mL of an ADC composition solution.

[0747] 4) The sample was filtered through a 0.22 um membrane.

[0748] (4) ADC purification using a Hitrap desalting column

[0749] The Hitrap desalting column was used after pretreatment in the following manner.

[0750] 1) Connect the Hitrap desalting column to the FPLC equipment.

[0751] 2) Sterilize the column by applying 20 mL of 0.5 M NaOH onto the resin, wash with distilled water, and stabilize the column with 20 mM histidine / histidine HC1, 8% sucrose pH 5.5.

[0752] 3) Inject the solution prepared in 1) into the FPLC.

[0753] 4) After purifying the antibody-drug conjugate, concentrate it using Ami con (50 KDa, 15 mL).

[0754] (5) DAR measurement using LC-MS LC-MS analysis was performed using an Agi lent 1260 / 1290 series HPLC system and TOF / Q-TOF mass spectrometry or Waters Xevo G3 Q-TOF mass spectrometry. DAR was calculated based on the peak intensity of the deconvoluted mass. 0.1 M DTT was added to the antibody-drug conjugate solution. The mixture was incubated at 37°C for 15-30 min to prepare a sample in which the disulfide bonds between the heavy and light chains of the antibody-drug conjugate and between the heavy chains were cleaved. The sample thus obtained was used for LC-MS analysis.

[0755] (6) Aggregate measurement using SEC-HPLC

[0756] Size-exclusion chromatography was performed at 25 °C using an Agilent 1260 series HPLC system with a TSK gel G3000SWXL size-exclusion chromatography column (7.8 X 300 mm, 5 μm). The mobile phase used was 78 mM KH2PO4, 122 mM K2HPO4, 250 mM KCl, 15% IPA, pH 7.0 ± 0.1. The flow rate was set at 0.75 mL / min. A sample of 40 - 50 μL was loaded per injection. Compounds were detected at 280 nm and 220 nm using a UV detector. The retention time of the aggregate peak was determined based on the molecular weight of the peak, and the degree of aggregation was determined by the relative area at 280 nm.

[0757] (7) Analysis of trastuzumab-linker-payload The results of the analysis of the prepared trastuzumab-linker-payload are shown in Table 4 below. [Table 4] Results of trastuzumab-linker-payload conjugation

[0758] | Reduced | |

[0759] …… …、… 三 | SEC-purity(%) | s Linker > TCEP / Ab : Drug / mAb > MS-DAR : 서

[0760] W | | | | | | s pay load ; ratio 서 ratio ; 서 서

[0761] <4. Effect of antibody-drug conjugate >

[0762] <Experimental Example 4> HER2 Extracellular Domain Competitive ELISA The antigen binding ability of the produced ADC was evaluated using HER2 ECD competitive ELISA (Acrobiosystems, EHM-VI). The experiment was performed according to the manual provided by the manufacturer. First, the ECD (extracellular domain) antigen of human HER-2 protein was dispensed into a 96-well plate at a concentration of 0.1 pg / ml, and the plate was sealed and coated at 4°C for 16 hours. The next day, the plate was washed three or more times with phosphate-buffered saline (PBS-T) containing 0.05% Tween-20, and then incubated at 37°C for 1.5 hours in a blocking solution containing 2% BSA in PBS-T to minimize nonspecific binding.

[0763] Biot inylated Ant i-HER-2 antibody was prepared at 0.05 pg / ml using a dilution solution and mixed with the standard substance and ADC sample in a 1:1 ratio. The mixture was then dispensed into each well to allow the coated human HER-2 ECD to competitively bind to the biot inylated ant i-HER-2 antibody or ADC. Then, Streptavidin-HRP and TMB substrate solutions at a concentration of 0.1 pg / ml were added in sequence and reacted for 1 hour and 20 minutes in a light-shielded state. Washing was performed three times with washing solution at each step, and finally, the absorbance value at the 450 band was measured using a plate reader. The stronger the binding strength between the antigen and the test substance, the lower the absorbance value because the competitive binding with the biot inylated Ant i-HER-2 antibody decreases. Therefore, the stronger the binding ability, the lower the absorbance value. The experimental results are shown in Figures 1 to 3, and it was confirmed that the produced ADC maintained antigen binding ability similar to trastuzumab.

[0764] <Experimental Example 5> Internalization Assay The degree of internalization of ADC produced in target expression cell lines was evaluated using a FACS Lyric flow cytometer from BD biosciences. The pH Sensor Labe 1 ing Kit (Doj indo, A558) was used according to the manual provided by the manufacturer to label the test substance with a pH-sensitive fluorescent dye. 100,000 SK-0V-3 cells were distributed per well in a 24-well plate, and the attachment of cells was confirmed the next day. The labeled test substance was diluted in the culture medium to a final concentration of 20 pg / ml, treated with the drug, and cultured for 24 hours. After removing the culture medium of the drug-treated cells, the cells were dissociated into single cells by treating with Accutase (Millipore, SCR005), and then centrifuged. After removing the supernatant and adding 500 pl of Cell Staining Buffer, the degree of internalization was analyzed using a FACS Lyric flow cytometer, and the degree of internalization was quantified as the percentage of APC-Cy7 positive cells, which is shown in Fig. 4. As a result, it was confirmed through the FACS experiment that trastuzumab and the produced ADC were internalized into cells at the same level, which means that the ADC of the present invention can effectively deliver the payload by internalizing into target cells.

[0765] <Experimental Example 6> In vitro anticancer efficacy of the ADC produced by in vitro cell proliferation assay was evaluated using Cel ITiter-Glo TM(Promega). Cells were cultured in RPMI-1640 medium containing 10% FBS. Cells were dispensed at a density of 2,500 cells per well in white 96-well plates and cultured for 24 hours. After treatment with drugs by serially diluting free payload or ADC in cell culture medium, the cells were cultured with the test substances for an additional 6 days at 37°C and 25% CO. After 6 days, cell growth inhibition was assessed by adding the same volume of CellTiter-Glo solution per well, reconstituted according to the manufacturer's instructions, to all test wells, incubating for 10 minutes, and measuring luminescence using a Spark® Multimode Microplate Reader TECAN instrument. Cell viability (%) was determined relative to the untreated control group as 100%. The results were plotted as a function of compound concentration using GraphPad Prism 9 software to calculate IC5O (50% inhibition of cell proliferation) values. The IC5o values ​​for each ADC are shown in Table 5 below.

[0766] [Table 5]

[0767]

[0768] A: CC50 <1 nM (less than 1 nM)

[0769] B: 1 <CC5o <10 nM(l 초과 10 nM 이하)

[0770] C: CC50 >1 pM (exceeding 1 pM) As can be seen in the table above, the ADC of the present invention was confirmed to have excellent cancer cell killing ability even at low concentrations against gastric cancer, breast cancer, and ovarian cancer cell lines. In particular, it was observed that it exhibited a specifically superior inhibitory effect on high-expression cell lines NCI-N87, SK-BR-3, and SK-OV-3, which express a large amount of the target antigen HER-2, compared to the negative cell line MDA-MB-231, thereby confirming that the ADC of the present invention exhibits an excellent anti-tumor effect on target cells expressing HER2, and can be excellent in the prevention or treatment of various diseases, such as cancer.

[0771] <Experimental Example 7> Evaluation of antitumor efficacy of antibody-drug conjugate in NCI-N87 xenograft model To evaluate the in vivo antitumor efficacy of the manufactured antibody-drug conjugate, efficacy evaluation was conducted in the NCI-N87 xenograft model.

[0772] Athymic Balb / C nude mice of 6-8 weeks of age were purchased and acclimatized for 7 days. Then, human gastric cancer cell line NCI-N87 (ATCC, CRL-5822) was diluted in a suspension of 1:1 mixed with PBS and Matrigel and injected subcutaneously on the right flank of the mouse at a dose of 5 x 10 per mouse. 6 Cells were injected. When the average tumor size after transplantation reached 80 to 120 mrf, mice were selected and randomly assigned to each group of 5 mice. Test substances, including the control group, were administered intravenously on the first day, and tumor size and weight were measured twice a week. Tumor volume was calculated according to the following formula to evaluate the tumor growth inhibition ability of the test substance. Tumor volume = ' T(a=short axis, b=long axis). The results are shown in Figs. 5 and 6. As can be seen in Fig. 5, the antibody-drug conjugate not only had a very excellent antitumor effect compared to the control group and the antibody-only administration group, but also showed a superior effect compared to Enhertu at the same dose. In addition, the control ADC in which linker-payload 6 was linked to the isotype control IgG did not have significant antitumor inhibitory activity, which means that the tested ADC showed a tumor inhibitory effect by target-dependently translocating to tumor tissues by the HER-2 antibody trastuzumab. In addition, no significant body weight loss was observed following the administration of any test substance used in the test (Fig. 6). Therefore, it was confirmed that the antibody-drug conjugate of the present invention exhibited excellent antitumor activity.

[0773] <Experimental Example 8> Evaluation of antitumor efficacy of antibody-drug conjugate in Calu-3 xenograft model To confirm the in vivo antitumor efficacy of the manufactured antibody-drug conjugate, efficacy was evaluated in the Calu-3 xenograft model. The overall experimental process was the same as Experimental Example 7. Specifically, Calu-3 (ATCC, HTB55), a human-derived non-small cell lung cancer cell line, was diluted in PBS and injected subcutaneously into the right flank of mice at a concentration of 10 x 10 per individual. 6Cells were injected. After group separation, test substances including control substances were administered once a week for a total of three times, and the results of monitoring tumor volume and animal body weight during the experimental period are shown in FIGS. 7 and 8. As a result, it was found that the antibody-drug conjugate of the present invention had an excellent antitumor effect equivalent to Enhertu in the Calu-3 non-small cell lung cancer model. Therefore, it was confirmed that the antibody-drug conjugate of the present invention showed excellent antitumor activity in lung cancer and could be excellent in the prevention or treatment of various diseases, such as cancer. <Experimental Example 9> Evaluation of antitumor efficacy of antibody-drug conjugate in NCI-N87 Enhertu resistant xenograft model In order to confirm the antitumor efficacy of the manufactured antibody-drug conjugate in the Enhertu resistant model, efficacy evaluation was conducted in the NCI-N87 Enhertu resistant xenograft model.

[0774] Enhertu resistant NCI-N87 cell line (Wuxi) was generated at Wuxi APPTEC by long-term treatment of 100 ng / ml Enhertu from NCI-N87 (ATCC, CRL-5822) parental cells. 6-8 week old NOD / SCID mice were purchased and acclimatized for 7 days. Enhertu resistant NCI-N87 was diluted in a suspension of 1:1 mixture of PBS and Matrigel, and 8X10 per mouse were injected intraperitoneally. 6cel Is was injected subcutaneously into the right flank of the mouse. When the average tumor size after transplantation reached 120 to 150 mrf, the mice were selected and randomly assigned to each group of 5 mice. The test substances including the control group were administered intravenously on the first day, and the results of monitoring the tumor volume and animal body weight during the experimental period are shown in Figs. 9 and 10. In summary, the antibody-drug conjugate of the present invention exhibited effective anticancer effects even in a model with Enhertu resistance. Enhertu, which showed excellent efficacy in the parental cell line NCI-N87 (Experimental Example 7), did not show significant efficacy in the current experimental example using NCI-N87 Enhertu R cells due to the acquired resistance mechanism of the resistant cell line, and the tumor continued to grow in the same manner as the control group, while the antibody-drug conjugate of the present invention was confirmed to have an excellent tumor inhibition effect at the same dose as Enhertu. On the other hand, no significant signs of toxicity were observed in observed items such as body weight changes. These results suggest that the ADC of the present invention overcomes the enhertu resistance mechanism and has the potential to treat patients who do not respond to existing treatments or who have relapsed.

[0775] <Experimental Example 10> ADC Characteristics Evaluation

[0776] (1) E. coli p-Glucuronidase Cleavage Test Test Method: The enzymatic cleavage of the p-glucuronide linker was evaluated by treating the cysteine derivatives of linker-payload 9 and linker-payload 11 with p-glucuronidase. In this test, commercially available E. coli-derived p-glucuronidase (Sigma, E.C.3.2.1.31) was used instead of the human-derived enzyme. Through this, it was possible to confirm whether the compounds of Example 3 and the compound of Example 43 were released and whether stable intermediates were formed during the reaction process. 12.5 μL of a 100 mM cysteine solution and 12.5 μL of a 30 mM borate buffer solution at pH 9 were added to 90 μL of purified water. Subsequently, 10 μL of linker-payload dissolved in a 10 mM DMSO solution was added. After 5 minutes, HPLC analysis results confirmed that all linker-payloads were completely converted to cysteine-linker-payload (cys-1 inker-pay load). 100 μL of the prepared cysteine-linker-payload solution was added to 880 μL of PBS, and then 20 μL of E. coli-derived p-glucuronidase (Sigma, E.C.3.2.1.31 Type IX-A, 1 mg / mL PBS solution) was added. The reaction mixture was reacted at 37 °C, and samples of 40 μL were taken at the time points of t = 0, 5, 10, 30, 60, and 90 minutes, respectively. 100 μL of acetonitrile was added to each sample, stirred for about 10 seconds, and then at 4 °Centrifugation was performed at 15,000 g for 10 minutes in In. The supernatant was used for quantitative analysis of cysteine-linker-payload and payload using LC-MS / MS, and the analysis was performed using a method similar to that described in U.S. Patent No. 8,568,728 (incorporated herein by reference). As a result of the test, it was confirmed from Figure 11 that the payload was rapidly released from the cysteine-linker-payload through a 1,6-elimination reaction following an enzymatic reaction by p-glucuronidase.

[0777] (2) Cathepsin L cleavage test experimental method:

[0778] Enzymatic cleavage of the GGFG (Glycine-Glycine-Phenylalanine-Glycine) linker was evaluated by treating trastuzumab-linker-payload 6 with human liver-derived cathepsin L (Sigma: EC 3.4.22.15). This allowed us to confirm the release of the desired drug, compound of Example 43, and the presence of stable intermediates generated during the reaction. Prior to use, cathepsin L (Sigma, Cat# C6854) was activated in cathepsin L digestion buffer (30 mmol / L dithiothreitol, pH 5.0) and then diluted to a final concentration of 120 nmol / L for use in the reaction. The reaction mixture was incubated at 37°C, and t

[0779] = 40 samples were collected at 0, 10, 30 minutes, 1, 2, 4, 6 and 24 hours, respectively. 100 pL of acetonitrile containing an internal standard was added to each sample. The mixture was stirred for about 10 seconds, then centrifuged at 4°C and 15,000 g for 10 minutes to secure the supernatant. The compound of Example 43 was quantitatively analyzed using LC-MS / MS. As a result of the test, it was confirmed from Figure 12 that the compound of Example 43 was rapidly released from the ADC through a cleavage reaction after the enzymatic reaction by cathepsin L.

[0780] (3) Cathepsin B cleavage test Test method:

[0781] Enzymatic cleavage of the VC (Vainine-Citrul line) linker was evaluated by treating the trastuzumab-linker-payload 10 with cathepsin B. In this study, commercially available recombinant cathepsin B (Sino Biological, EC3.4.22.1) was used instead of the human enzyme. This allowed us to confirm whether the target drug, compound of Example 3, was released and whether a stable intermediate was formed during the reaction. Before use, cathepsin B (Sino Biological, Cat#10483- H08H) was kept in a cooled solution at pH 5.0.

[0782] After activation using cathepsin B digestion buffer (30 mmol / L DTT), it was diluted to a final concentration of 1 U for reaction. The reaction mixture was reacted at 37°C, and 40 pL samples were collected at t = 0, 10, 30 min, 1, 2, 4, 6, and 24 h, respectively. 100 pL of acetonitrile containing an internal standard was added to each sample, stirred for about 10 seconds, and centrifuged at 4°C and 15,000 g for 10 minutes. The supernatant was used for quantitative analysis of the compound of Example 3 using LC-MS / MS. As a result of the test, it was confirmed from Figure 13 that the compound of Example 3 was rapidly released from the ADC through a cleavage reaction following the enzymatic reaction by cathepsin B.

[0783] (4) Stability test of ADC (Antibody-drug conjugate) in plasma and buffer solution Experimental method: To evaluate the stability of antibody-drug conjugate (ADC) in plasma and phosphate-buffered saline (PBS) containing 1% BSA (bovine serum albumin), trastuzumab-linker-payload 8, trastuzumab-linker-payload 9, and trastuzumab-linker-payload 10 were prepared at a concentration of 100 pg / mL, respectively. Each solution was incubated at 37°C for 14 days, and samples were collected on days 0, 1, and 3 or 4, 7, and 14. For total antibody quantitative analysis, 20 pL of each sample was added to PBS containing 0.1% Tween 20.

[0784] 200 pL of 10 pL protein A magnetic beads were mixed. The sample was gently reacted for about 1 hour at room temperature, then fixed on a magnetic rack and washed with 200 pL of PBS containing 0.1% Tween 20. The washed beads were resuspended in 50 mM Tris-HCl buffer (pH 8.0).

[0785] It was resuspended in 100 pL. 4|』L of 500 mM DTT (dithiothreitol) was added, and the reaction was performed at 70°C for about 10 minutes. After cooling to room temperature, 1pL of trypsin solution (1 mg / mL) was added, and the digestion reaction was induced at 37°C for about 24 hours. After that, 30 pL of acetonitrile containing the internal standard was added, stirred for about 10 seconds, and centrifuged at 4°C for 15,000 × g for 10 minutes. The supernatant was collected, transferred to an LC vial, and analyzed by LC-MS / MS. As a result, the stability of all ADCs was confirmed to be about 80% at the 14-day point (Figs. 14 to 16).

[0786] (5) Pharmacokinetics of trastuzumab-linker-payload 9 and trastuzumab-linker-payload 10 in rat plasma Method: The pharmacokinetic characteristics of trastuzumab-linker-payload 9 and trastuzumab-linker-payload 10 in plasma were evaluated after a single intravenous administration (IV bolus, 3 mg / kg) of each antibody-drug conjugate (ADC) to male SD rats. Blood was collected at 1, 2, 4, 8, 24, 48, 96, 168, and 336 hours after administration, and the concentration of ADC in plasma was measured using the same method as in the plasma stability test. After a single intravenous dose of 3 mg / kg, the total antibodies of trastuzumab-linker-payload 9 and trastuzumab-linker-payload 10 exhibited the following pharmacokinetic characteristics: plasma clearance (CL) was 4.6±0.1 and 6.3±0.9 mL / day / kg, respectively; volume of distribution (Nz) was 90.8±8.3 and 93.8±3.5 mL / kg, respectively; terminal half-life (T1 / 2) was 13.6±1.1 and 10.5±1.9 days, respectively; initial plasma concentration (C1 / 2) was 1.5±1.1 and 1.5±1.9 days, respectively; and initial plasma concentration (C1 / 2) was 1.5±1.1 and 1.5±1.9 days, respectively. o ) were 81.6±8.4 and 92.1±8.0 pg / mL, respectively, and the area under the plasma concentration-time curve (AUCo- iast) from administration to the last quantifiable time point was 347.1±16.2 and 302.7±13.9 daypg / mL, respectively. The LC-MS / MS analysis results of ADC are shown in Fig. 17 and Table 6.

[0787] [Table 6]

[0788] (6) Pharmacokinetics of trastuzumab-linker-payload 5, trastuzumab-linker-payload 6, and trastuzumab-linker-payload 11 in rat plasma Method: The pharmacokinetic characteristics of trastuzumab-linker-payload 5, trastuzumab-linker-payload 6, and trastuzumab-linker-payload 11 in plasma were evaluated after a single intravenous administration (IV bolus, 3 mg / kg) of each antibody-drug conjugate (ADC) to male SD rats. Blood was collected at 1, 2, 4, 8, 24, 48, 96, 168, and 336 hours after administration, and the concentration of ADC in plasma was measured using the same method as in the plasma stability test.

[0789] After a single intravenous dose of 3 mg / kg, the total antibodies of trastuzumab-linker-payload 5, trastuzumab-linker-payload 6, and trastuzumab-linker-payload 11 exhibited the following pharmacokinetic characteristics: plasma clearance (CL) was 17.5±1.8, 3.6±0.3, and 4.1±0.3 mL / day / kg, respectively; volume of distribution (Nz) was 162.9±5.4, 66.8±6.7, and 91.2±8.5 mL / kg, respectively; terminal half-life (T1 / 2) was 6.5±0.5, 12.8±2.4, and 15.4±2.3 days, respectively; initial plasma concentration (C1 / 2) was 1.5±0.5, 1.5±0.5, and 1.5±0.5 days, respectively. o ) were 40.9±1.2, 94.7±6.7, and 100.3±10.4 pg / mL, respectively, and the area under the plasma concentration-time curve (AUCo-iast) from administration to the last quantifiable time point was 124.2±29.2, 450.4±19.0, and 359.1±9.0 day .pg / mL, respectively. The LC-MS / MS analysis results of ADC are shown in Fig. 18 and Table 7.

[0790] [Table 7]

[0791] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without changing the technical idea or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. The scope of the present invention should be interpreted as including within the scope of the present invention all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts rather than the above detailed description.

Claims

【Scope of Claims】 【 Claim 11 An immunoconjugate represented by the following formula I or a pharmaceutically acceptable salt thereof: [Formula I] Ab-(L-D) p In Formula I above, 日 is a cytotoxic drug moiety represented by the following Formula III, [Chemical Formula III] In the above formula III, Ri is pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, pyrrolyl, "4, bucket:, :, = r 1 / jo A ~ if s f new Y furanyl, phenyl, phenolic, "", ;, At this time, the inside of the above-mentioned day is unsubstituted or at least one of -H7} is each independently -NH2, -(C1-3alkyl)NH2, -NH(C1-3alkyl), -N(C1-3alkyl)2, -OH, -NO2, C1-5alkyl, -F, -Cl, -Br, or -1 substituted, R2 is pyridinyl, pyrimidinyl, phenyl, piperidinyl, piperazinyl, morpholinyl, pyrazolyl, imidazolyl, phenolic, - CH2C(=0)NH2, - C(=0)NH2, - C(=0)NHCH2CH3, - CH2NHC(=0)CH2CH3, -NH2, -NHCH3, -NHCH2CH2CH3, At this time, the -H of the above R2 is each independently unsubstituted or at least one of -H is each independently -NH2, -NH(C1-5alkyl), -M(C1-5alkyl)2, -NHCH3, -N(CH3)2, -NHCH2CH3, -OH, -NO2, -S(=O)2CH3, -C1-5alkyl (the inside of the alkyl is unsubstituted or at least one of the inside is each independently substituted with one selected from the group consisting of phenyl, pyridinyl, morpholinyl, piperazinyl and piperidinyl), -(C1-5alkyl)NH2, -morpholinyl, -piperidinyl, -piperazinyl, -phenyl, -pyridinyl, -pyrimidinyl (the inside of the above -morpholinyl, -piperidinyl, -piperazinyl, -phenyl, -pyridinyl and -pyrimidinyl is unsubstituted or at least one of the inside is each independently substituted with one selected from the group consisting of -C1-3alkyl and -CF3), -cyclohexyl, -cyclobutoxy, -cyclopropoxy, -cyclopentoxy, -F, -Cl, -Br, -I, -CHF2, -CF3, -C1-3alkoxy, - NHC(=O)CH3, -C(=O)CH2CH3, -C(=O)N(CH3)2, -C(=O)NH(CH3), -C(=O)NH2, -S(=O)2NHCH3; - S(=0)2, - S(=0)2CH3, -trifluoromethylphenyl, (The above Ray is -H, - C1-5 alkyl or - CF3), is substituted, Za and Zb are each independently one selected from the group consisting of - visible or the following 1) to 18), 1) Ci-5 alkyl; 2) -F, -Cl, -Br, or -I; i 2, - CH2OH), (wherein Rb2 is - CH『, -NH-, or - 0-, and Rb3 and Rb』 are each independently - H, - NH2, -F, -Cl, -Br, or -I), pyrimidine or pyrrole, and Rx9 is - H, or Ci-5 alkyl; 5) -COOH; 6) -NRxioRxn, where Rxio or Rx n are each independently - H, or - Ofe; 7) -CFs; 8) -CN; 9) morpholine or morpholine substituted with one or more of each independently Ci-5 alkyl, - F, -C1, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2; 10) piperidine or piperidine substituted with one or more - Hs each independently with C1-5 alkyl, - F, - Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2; 11) piperazine or piperazine substituted with one or more -H7} each independently with Ci-5 alkyl, - F, - Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or - C(=0)CH=CH2; 12) -0Rxi2, where Rxi2 is -CFs or C1-3 alkyl; 13) -0H； 14), where 거 is 0, 1, 2 or 3, and Rxi3 is - NH2, - NH(CHs), -N(CH3)2, methylpyrazolyl, pyrazolyl, piperazine, piperidine, or morpholine; 327 15), where RX14 is -CH2-, -NH-, or - 0-, and RX15 is - H, straight or branched Ci-5 alkyl, - NH2, -F, -Cl, -Br, or - 1; r '처 H ..ah. i:: 3 "、、、,、 16) ° 1 Huh— , where RX16 is C1-5 alkyl or - CH2CH2N(CH3)2 ; 5.. Dou 제 '••'''dou L J 17); or 18) - (C1-3 alkyl)- NH2 Ak is - CH『 , - (CH2)『 , - (CH2)3 - or , - CH『 , _ (CH2)2 _ , - [[ID==19]](CH2)3 - or each of 내 is independently unsubstituted or at least one of 내 is independently substituted with - NH2, -0H, - N02, -C1-5 alkyl, - CH2NH2, -CFs, - OCF3, -CN, -F, -Cl, -Br, or -I; L is a linker moiety connecting Ab오}日; Ab is an antibody or an antigen-binding fragment thereof; and n is an integer from 1 to 10. [

2. ] In the previous claim, R is pyridine, pyrazole, imidazole, phenyl, or At this time, the inner side of the Ri is not substituted or at least one of the inner sides is independently substituted with - NH2, -OH, -N02 or - F, At this time, each of the Rs in R2 is independently unsubstituted or at least one of them is independently -NH2, -C1-5alkyl, -(C1-5alkyl)NH2, -morpholine, 329 -piperazinyl, substituted, Za and Zb are each independently -O- or one selected from the group consisting of the following 1) to 6), 1） -F； Here, c is 0, 1, 2 or 3, and RX8 is piperidine, g triple-'hole 허 'small H , Rb! (The above Rbi is - NH2), or (the above Rb2 is -CH2- or -NH-, and Rb3 and Rb4 can each independently be -H or -NH2); and RX9 is as follows; 3) —NRx10Rx11 where Rx10 and Rx11 are both as follows; 4) Piperidinyl; ...,...,% of the hole 4iRX ⑯ 5) , where RX14 is - CH2 - and RX15 is - H or - NH2; 6), where RX16 is C1-5alkyl, Ak is -CH2-, -(CH2)2 - or -(CH2)3 -, an immunoconjugate or a pharmaceutically acceptable salt thereof. 【 In claim 3], n is pyridine or, R2 is pyridine or phenyl, where one moiety of phenyl is substituted with piperazine, Za and Zb are each independently - H, -F, Here, c is 2, (where Rb2 is -NH-, and Rb3 and Rb4 are both -H); and RX9 is as follows; Ak is -CH2- or -(CH2)2 -, an immunoconjugate or a pharmaceutically acceptable salt thereof. 【 Claim 4 331 In claim 1, the cytotoxic drug moiety represented by the chemical formula III is an immunoconjugate or a pharmaceutically acceptable salt thereof, which is any one of the compounds of Examples 1 to 43 below: 【 Claim 5 According to claim 1, the cytotoxic drug moiety represented by the formula III is a compound represented by the following formula Any one of IV-1 to IV-4: an immunoconjugate or a pharmaceutically acceptable salt thereof. 341 [Chemical Formula IV-1] 【 Claim 6 According to claim 1, L includes any one or more of the following a) to d): an immunoconjugate or a pharmaceutically acceptable salt thereof: a) a cleavable unit; b) a dipeptide, tripeptide or tetrapeptide unit; c) a glucuronide unit; and d) a PEG unit. 【 Claim 7 According to claim 1, L is represented by the following formula V: an immunoconjugate or a pharmaceutically acceptable salt thereof: [Chemical Formula V] , Ly is a single bond or -valine-citrulline- or -glycine-glycine-phenylalanine-glycine-; and Lx and Ly cannot both be a single bond at the same time; Lz is a PABC (para-aminobenzyl carbamate), PAB (para-aminobenzyl), or glucuronide moiety; e is an integer from 1 to 6; h and j are each independently an integer from 1 to 3; at least two of f, g, and i are 0, and when f, g, or i are not 0, f or g is an integer from 2 to 16, and i is an integer from 16 to 30. 【

8. In claim 1, L is an immunoconjugate represented by any one of the following chemical formulae V-1 to V-10, or a pharmaceutically acceptable salt thereof: [Chemical Formula V-1] [Chemical Formula V-5] [Chemical Formula V-9] 346 【

9. In claim 1, L- D is an immunoconjugate represented by any one of the following chemical formulas VI-1 to VI-11, or a pharmaceutically acceptable salt thereof: [Chemical Formula VI-1] 347 [Chemical Formula VI-4] [Chemical Formula VI-7] 349 [Chemical Formula VI-11]

10. In claim 1, the antibody is an immunoconjugate or a pharmaceutically acceptable salt thereof that targets a cancer cell-specific antigen. 【 청구항 11】 제 10항에 있어서, 암세포 특이적 항원은 5T4, ABL, ABCF1, ACVR1, ACVR1B, ACVR2, ACVR2B, ACVRL1, AD0RA2A, AFP, Aggrecan, AGR2, AICDA, AIF1, AIGI, AKAP1, AKAP2, ALCAM, ALK, AMH, AMHR2, ANGPT1, ANGPT2, ANGPTL3, ANGPTL4, ANPEP, APC, APOCI, AR, 아로마타제 (aromatase), ASPH, ATX, AX1, AXL, AZGP1(zinc-a- glycoprotein), B4GALNT1, B7, B7.1, B7.2, B7-H1, B7-H3, B7-H4, B7-H6, BAD, BAFF, BAG1, BAI1, BCR, BCL2, BCL6, BCMA, BDNF, BLNK, BLR1 (MDR15), BlyS, BMP1, BMP2, BMP3B (GDFIO), BMP4, BMP6, BMP8, BMP10, BMPR1A, BMPR1B, BMPR2, BPAG1 (플렉틴), BRCA1, C19orfl0 (IL27w), C3, C4A, C5, C5R1, CA6, CA9, CANT1, CAPRIN-1, CASP1, CASP4, CAV1, CCBP2 (D6 / JAB61), CCL1 (1-309), CCLI1 (에오탁신), CCL13 (MCP-4), CCL15 (MIP-Id), CCL16 (HCC-4), CCL17 (TARC), CCL18 (PARC), CCL19 (MIP- 3b), CCL2 (MCP-1), MCAF, CCL20 (MIP- 3a), CCL21 (MEP-2), SLC, exodus-2, CCL22(MDC / STC-I), CCL23 (MPIF-I), CCL24 (MPIF- 2 / 에오탁신- 2) , CCL25 (TECK) , CCL26(에오탁신- 3), CCL27 (CTACK / ILC) , CCL28, CCL3 (MIP-Ia), CCL4 (MIPIb), CCL5(RANTES) , CCL7 (MCP-3), CCL8 (mcp-2) , CCNA1 , CCNA2, CCND1 , CCNE1 , CCNE2, CCR1 (CKR1 / HM145) , CCR2 (mcp-IRB / RA) , CCR3 (CKR3 / CMKBR3) , CCR4, CCR5(CMKBR5 / ChemR13) , CCR6 (CMKBR6 / CKR-L3 / STRL22 / DRY6) , CCR7 (CKR7 / EBI1), CCR8 또는 CDwl98 (CMKBR8 / TERI / CKR-L1) , CCR9 (GPR-9-6) , CCRL1 (VSHK1), CCRL2 (L-CCR) , CD13, CD164, CD19, CDH6, CDIC, CD2, CD20, CD21, CD200, CD22, CD23, CD24, CD27, CD28, CD29, CD3, CD33, CD35, CD37, CD38, CD3E, CD3G, CD3Z, CD4, CD40, CD40L, CD44, CD45RB, CD47, CD52, CD56, CD69, CD70, CD72, CD74, CD79A, CD79B, CD8, CD80, CD81, CD83, CD86, CD97, CD99, CD117, CD125, CD137, CD147, CD179b, CD223 , CD279, CDH1 (E-카드헤린), CDH10, CDH12, CDH13 , CDH18, CDH19, CDH20, CDH3, CDH5, CDH7, CDH8, CDH9, CDH17, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK9, CDKN1A (p21Wapl / Cipl) , CDKN1B (p27Kipl), CDKN1C, CDKN2A (pl6INK4a) , CDKN2B, CDKN2C,CDKN3, CEA, CEACAM5, CEACAM6, CEBPB, CERI, CFC1B, CHGA, CHGB, 키티나제 (Chitinase), CHST10, CIK, CKLFSF2, CKLFSF3, CKLFSF4, CKLFSF5, CKLFSF6, CKLFSF7, CKLFSF8, CLDN3, CLDN6, CLDN7 (클라우딘- 7), CLDN18, CLEC5A, CLEC6A, CLEC11A, CLEC14A, CLN3, CLU (클러스테린), CMKLR1, CMK0R1 (RDC1), CNR1, C-MET, C0L18A1, COL I Al, C0L4A3, C0L6A1, CR2, Crypto, CRP, CSF1 (M-CSF), CSF2 (GM-CSF), CSF3 (GCSF), CTAG1B (NY-ESO-1), CTLA4, CTL8, CTNNB1 (b-카테닌), CTSB (카텝신 B), CX3CL1 (SCYD1), CX3CR1 (V28), CXCL1 (GR01), CXCL10 (IP-IO), CXCLI1 (l-TAC / IP-9), CXCL12 (SDF1), CXCL13, CXCL14, CXCL16, 352 CXCL2 (GR02), CXCL3 (GR03), CXCL5 (ENA-78 / LIX), CXCL6 (GCP-2), CXCL9 (MIG), , EFNB2, EGF, EGFR, ELAC2, ENG, Enola, EN02, EN03, EpCAM, EPHA1, EPHA2, EPHA3, EPHA4, EPHA5, EPHA6, EPHA7, EPHA8, EPHA9, EPHA10, EPHB1, EPHB2, EPHB3, EPHB4, EPHB5, EPHB6, EPHRIN-A1, EPHRIN-A2, EPHRINA3, EPHRIN-A4, EPHRIN-A5, EPHRIN-A6, EPHRIN-B1, EPHRIN-B2, EPHRIN-B3, EPHB4, EPG, ERBB2 (HER-2), ERBB3, ERBB4, EREG, ERK8, 에스트로겐 수용체 (Ear 1과 ESR2) , F3 (TF) , FADD, FAP, 파르네실트란스퍼라제 , FasL, FASNf , FCER1A, FCER2, FCGR3A, FGF, FGF1 (aFGF), FGF10, FGF11, FGF12, FGF12B, FGF13, FGF14, FGF16, FGF17, FGF18, FGF19, FGF2 (bFGF), FGF20, FGF21, FGF22, FGF23, FGF3 (int-2), FGF4 (HST), FGF5, FGF6 (HST-2), FGF7 (KGF), FGF8, FGF9, FGFR1 , FGFR2, FGFR3 , FGFR4, FIGF (VEGFD), FILl(EPSILON), FBL1 (ZETA), FL J 12584, FLJ25530, FLRT1 (피브로넥틴), FLT1, FLT-3, FOLR1(FR—알파), FOS, FOSLl(FRA-l) , FY(DARC) , GABRP(GABAa) , GAGEB1 , GAGECI ,GALNAC4S-6ST, GATA3, GD2, GD3, GDF5, GFI1, GFRA1, GGT1, GM-CSF, GNAS1, GNRH1, GPC1, GPC3, GPNB, GPR2 (CCR10), GPR31, GPR44, GPR81 (FKSG80), GRCC10 (CIO), GRP, GSN (Gelsol in), GSTP1, GUCY2C, HAVCR1, HAVCR2, HDAC, HDAC4, HDAC5, HDAC7A, HDAC9, Hedgehog, HGF, HIF1A, HIP1, histamine and histamine receptor, HLA- A, HLA-DR, HLA-DRA, HLA-E, HM74, HMOXI, HSP90, HUMCYT2A, ICEBERG, ICOSL, ID2, IFN-a, IFNA1, IFNA2, IFNA4, IFNA5, EFNA6, BFNA7, IFNB1, IFN-gamma, IFNW1, IGBP1, IGF1, IGFIR, IGF2, IGFBP2, IGFBP3, IGFBP6, DL-1, ILIO, ILIORA, ILIORB, IL-1, IL1R1 (CD121a), ILlR2 (CD121b), IL-IRA, IL-2, IL2RA (CD25), IL2RB (CD122), 353 IL2RG(CD132) , IL-4, IL- 4R(CD123) , IL-5, IL5RA(CD125) , IL3RB(CD131) , IL-6, IL6RA, (CD126), IR6RB(CD130), IL-7, IL7RA(CD127), IL-8, CXCR1 (IL8RA), CXCR2, (IL8RB / CD128), IL-9, IL9R(CD129), IL-10, IL10RA(CD210), IL10RB(CDW210B), IL-11, IL11RA, IL-12, IL-12A, IL-12B, IL-12RB1, IL-12RB2, IL-13, IL13RA1, IL13RA2, IL14, IL15, IL15RA, IL16, IL17, IL17A, IL17B, IL17C, IL17R, IL18, IL18BP, IL18R1, IL18RAP, IL19, ILIA, ILIB, ILIF10, ILIF5, IL1F6, IL1F7, IL1F8, DL1F9, ILIHYI, ILIR1, IL1R2, ILIRAP, ILIRAPLI, ILIRAPL2, ILIRL1, IL1RL2, ILIRN, IL2, IL20, IL20RA, IL21R, IL22, IL22R, IL22RA2, IL23, DL24, IL25, IL26, IL27, IL28A, IL28B, IL29, IL2RA, IL2RB, IL2RG, IL3, IL30, IL3RA, IL4, 1L4, IL6ST (Glycoprotein 130), ILK, INHA, INHBA, INSL3, INSL4, IRAKI, IRAK2, ITGA1, ITGA2, ITGA3, ITGA6 (alpha6 integrin), ITGAV, ITGB3, ITGB4 (beta4 integrin), JAG1, JAK1, JAK3, JTB, JUN, K6HF, KAI1, KDR, KIT, KITLG, KLF5 (GC Box BP), KLF6, KLK10, KLK12, KLK13, KLK14, KLK15, KLK3, KLK4, KLK5, KLK6, KLK9, KRT1, KRT19 (Keratin 19), KRT2A, KRTHB6 (hair-specific type II keratin), L1CAM, LAG3, LAMA5, LAMP1, LEP (Leptin),Lewis Y antigen (LeY), LILRB1, Lingo— p75, Lingo- Troy, LGALS3BP, LRRC15, LPS, LTA (TNF-b), LTB, LTB4R (GPR16), LTB4R2, LTBR, ​​LY75, LYPD3, MACMARCKS, MAG or OMgp, MAGEA3, MAGEA6, MAP2K7 (c-Jun), MDK, MIB1, midkine, MIF, MISRII, MJP-2, MLSN, MK, MKI67 (Ki-67), MMP2, MMP9, MSMB, MT3 (metallothionein— UI), mTOR, MTSS1, MUC1 (mucin), MUC16, MYC, MYD88, NCK2, NCR3LG1, neurocan, NFKBI, NFKB2, NGFB (NGF), NGFR, NgR-Lingo, NgRNogo66, (Nogo) , NgR— p75, NgR-Troy, NMEI (NM23A) , NOTCH, N0TCH1 , N0TCH3 , N0X5, NPPB, NR0B1 , NR0B2, NRID1, NR1D2, NR1H2, NR1H3 , NR1H4, NR112, NR113, 354 NR2C1, NR2C2, NR2E1, NR2E3, NR2F1, NR2F2, NR2F6, NR3C1, NR3C2, NR4A1, NR4A2, NR4A3, NR5A1, NR5A2, NR6A1, NRP1, NRP2, NT5E, NTN4, NY-ES01, ODZI, OPRDI, P2RX7, PAP, PARTI, PATE, PAWR, P—gardherin, PCA3, PCD1, PD-L1, PCDGF, PCNA, PDGFA, PDGFB, PDGFRA, PDGFRB, PECAMI, Ll-CAM, peg—heart{sparaginase}, PF4 (CXCL4), PGF, PGR, phosphacan (phosphacan), PIAS2, PI3 kinase, PIK3CG, PLAU (uPA), PLG, PLXDCI, PKC, PKC—beta, PPBP (CXCL7), PPID, PR1, PRAME, PRKCQ, PRKD1, PRL, PROC, PR0K2, PSAP, PSCA, PSMA, PTAFR, PTEN, PTHR2, PTGS2 (COX— 2), PTN, PVRIG, RAC2 (P21Rac2), RANK, RANK ligand, RARE, RGS1, RGS13, RGS3, RNFI10 (ZNF144), Ron, R0B02, R0R1, RXR, S100A2, SCGB1D2 (lipophilin B), SCGB2A1 (mammaglobin 2), SCGB2A2 (mammaglobin 1), SCYE1 (endothelial monocyte-activating cytokine), SDF2, SERPENA1, SERPINA3, SERPINB5 (maspin), SERPINEI (PAI-I), SERPINFI, SHIP-1, SHIP-2, SHB1, SHB2, SHBG, SfcAZ, SLC2A2, SLC33A1, SLC43A1, SLC44A4, SLC34A2, SLIT2, SPP1, SPRR1B (Sprl), ST6GAL1, ST8SIA1, STAB1, STATE, STEAP, STEAP2, TB4R2, TBX21, TCP10, TDGF1, TEK,TGFA, TGFB1 , TGFB1I1 , TGFB2 , TGFB3 , TGFBI , TGFBR1 , TGFBR2 , TGFBR3 , THINGS, THBS1 (antigen- 1), THBS2, THBS4, THPO, TIE (Tie-1), TIMP3 , functional 인자 (tissue factor), TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11, TNF, TNF-a, TNFAIP2 (B94) , TNFAIP3, TNFRSFI1A, TNFRSF1A, TNFRSF1B, TNFRSF21 , . TNFRSF5, TNFRSF6 (Fas), TNFRSF7, TNFRSF8, TNFRSF9, TNFSF10 (TRAIL), TNFRSF10A, TNFRSF10B, TNFRSF12A, TNFRSF17, TNFSF11 (TRANCE) , TNFSF12 (AP03L) , TNFSF13 (Apr), TNFSF13B, TNFSF14 (HVEM- L), TNFRSF14 (HVEM) , TNFSF15 (VEGI), TNFSF18, TNFSF4 (0X40 °C) , TNFSF5, (CD40 ligand), TNFSF6 (FasL), TNFSF7 (CD27 ligand), TNFSF8 (CD30 ligand), 355 TNFSF9 (4-1BB ligand), TOLLIP, Toll-like receptor, T0P2A (topoisomerase Ila), TP53, TPM1, TPM2, TRADD, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5, TRAF6, TRKA, TREM1, TREM2, TR0P2, TRPC6, TSLP, TWEAK, Tyrosinase, uPAR, VEGF, VEGFB, VEGFC, Versican, VHL C5, VLA-4, WT1, Wnt-1, XCL1 (lymphotactin), XCL2 (SCM-Ib), XCRI (GPR5 / CCXCR1), YY1, ZFPM2, CLEC4C (BDCA-2, DLEC, CD303, CDH6, CLECSF7), CLEC4D (MCL, CLECSF8), CLEC4E (Mincle), CLEC6A (Dectin-2), CLEC5A (MDL-1, CLECSF5), CLEC1B (CLEC-2), CLEC9A (DNGR-1), CLEC7A (Dectin-1), CLEC11A, PDGFRa, SLAMF7, GP6 (GPVI), LILRA1 (CD85I), LILRA2 (CD85H, ILT1), LILRA4 (CD85G, ILT7), LILRA5 (CD85F, ILT11), LILRA6 (CD85b, ILT8), LILRB1, NCR1 (CD335, LY94, NKp46), NCR3 (CD335, LY94, NKp46), NCR3 (CD337, NKp30), OSCAR, TARM1, CD30, CD300C, CD300E, CD300LB (CD300B), CD300LD (CD300D), KIR2DL4 (CD158D), KIR2DS, KLRC2 (CD159C, NKG2C), KLRK1 (CD314, NKG2D), NCR2 (CD336, NKp44), PILRB, SIGLEC1 (CD169, SN), SIGLEC5, SIGLEC6, SIGLEC7, SIGLEC8, SIGLEC9, SIGLEC10,SIGLEC11, SIGLEC12, SIGLEC14, SIGLEC15 (CD33L3), SIGLEC16, SIRPA, SIRPB1 (CD172B), TREM1 (CD354), TREM2, KLRF1 (NKp80), 17- 1A, SLAM7, MSLN, CTAG1B / NY-ES0-1, MAGEA3 / A6, ATP5I (Q06185), OAT (P29758), AIFM1 (Q9Z0X1), AGFA (Q64133), MTDC (P18155), CMC1 (Q8BH59), PREP (Q8K411), YMEL1 (088967), LPPRC (Q6PB66), LONM (Q8CGK3), ACON (Q99KI0), 0D01 (Q60597), IDHP (P54071), ALDH2 (P47738), ATPB (P56480), AATM (P05202), TMM93 (Q9CQW0), ERG I 3 (Q9CQE7), RTN4 (Q99P72), CL041 (Q8BQR4), ERLN2 (Q8BFZ9), TERA (Q01853), DADI (P61804), CALX (P35564), CALU (035887), VAPA (Q9WV55), MOGS (Q80UM7) GANAB (Q8BHN3), ER01A (Q8R180), UGGG1 (Q6P5E4), P4HA1 (Q60715), HYEP (Q9D379), CALR (P14211), AT2A2 (055143), PDIA4 (P08003), PDIA1 (P09103), PDIA3 (P27773), PDIA6 (Q922R8), CLH (Q68FD5), PPIB (P24369), TCPG (P80318), M0T4 (P57787), NICA (P57716), BAS I (P18572), VAPA (Q9WV55), ENV2 (P11370), VAT1 (Q62465), 4F2 (P10852), ENOA (P17182), ILK (055222), GPNMB (Q99P91), ENV1 (P10404), ER01A (Q8R180) , CLH (Q68FD5) , DSG1A (Q61495) , AT1A1 (Q8VDN2) , HY0U1 (Q9JKR6) , An immunoconjugate or a pharmaceutically acceptable salt thereof, wherein the immunoconjugate is at least one selected from the group consisting of TRAP1 (Q9CQN1), GRP75 (P38647), ENPL (P08113), CH60 (P63038), and CH10 (Q64433).

12. In claim 1, the antibody is an immunoconjugate or a pharmaceutically acceptable salt thereof targeting HER2.

13. A pharmaceutical composition for preventing or treating cancer, comprising the immunoconjugate of any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof as an active ingredient.

14. A pharmaceutical composition for preventing or treating cancer, wherein the cancer is a cancer in which NAMPT is overexpressed or overactivated, according to claim 13.

15. A pharmaceutical composition for preventing or treating cancer, according to claim 13, wherein the cancer is HER2 positive or mutant cancer.

16. A method for preventing or treating cancer, comprising administering a therapeutically effective amount of an immunoconjugate of any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

17. Use of the immunoconjugate of any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof for preventing or treating cancer.

18. Use of the immunoconjugate of any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for preventing or treating cancer. 【Claim 1 is a drug-linker conjugate represented by the following chemical formula II or a pharmaceutically acceptable salt thereof: 358 [Chemical Formula II] In the above chemical formula 11, LD is a cytotoxic drug moiety represented by the following chemical formula III, [Chemical Formula III] In the above chemical formula III, Ri is pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, pyrrolyl, furanyl, phenyl, phenolic, , H7} are each independently substituted with -NH2, - (C1-3 alkyl) NH2, - NH(CI-3 alkyl), - MC1-3 alkyl) 2, -OH, - N02, Ci-5 alkyl, - F, -Cl, -Br, or -I, R2 is pyridinyl, pyrimidinyl, phenyl, piperidinyl, piperazinyl, morpholinyl, pyrazolyl, imidazolyl, phenolic, - CH2C(=0)NH2, - C(=0)NH2, - C(=0)NHCH2CH3, - CH2NHC(=0)CH2CH3, -NH2, -NHCH3, -NHCH2CH2CH3, At this time, the inside of the above R2 is independently unsubstituted or at least one of the insides is independently - NH2, - NH (CI-5 alkyl), - MC1-5 alkyl) 2, -NHCHs, -N (CH3) 2, - NHCH2CH3, -OH, - NO2, - S (=0) 2CH3, - C1-5 alkyl (the inside of the above alkyl is unsubstituted or at least one of the insides is independently substituted with one selected from the group consisting of phenyl, pyridinyl, morpholinyl, piperazinyl and piperidinyl), - (Ci-5 alkyl) NH2, -morpholinyl, -piperidinyl, -piperazinyl, -phenyl, -pyridinyl, -pyrimidinyl (wherein the -morpholinyl, -piperidinyl, -piperazinyl, -phenyl, -pyridinyl and -pyrimidinyl may be unsubstituted or at least one of them is independently substituted with one selected from the group consisting of -C1-3alkyl and -CF3), -cyclohexyl, -cyclobutoxy, -cyclopropoxy, -cyclopentoxy, -F, -Cl, -Br, -I, -CHF2, -CF3, -C1-3alkoxy, - NHC(=0)CH3, -C(=0)CH2CH3, -C(=0)N(CH3)2, -C(=0)NH(CH3) , -C(=0)NH2, -S(=O)2NHCH3; substituted with, Za and Zb are each independently -void or one selected from the group consisting of the following 1) to 18), 1) C1-5alkyl; 2) -F, -Cl, -Br, or -I; i 2, CH2OH), (wherein Rb2 is -CH2, -NH- or -O-, and Rb3 and Rb4 are each independently -H, -NH2, -F, -Cl, -Br, or -I), pyrimidinyl or pyrrolyl, and RX9 is -H, or -C1-5alkyl; 5) -COOH; 6) -NRxioRxn, where Rxio or Rx n are each independently - H, or - Ofe; 7) -CF3; 8) -CN; 9) morpholinyl or morpholinyl in which one or more of them are each independently substituted with C1-5alkyl, -F, -Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or -C(=0)CH=CH2; 10) piperidinyl or piperidinyl in which one or more -H are each independently substituted with C1-5alkyl, -F, -Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or -C(=0)CH=CH2; 11) piperazinyl or piperazinyl in which one or more -H are each independently substituted with C1-5alkyl, -F, -Cl, -Br, -I, -C(=0)NHCH3, -C(=0)CH3 or -C(=0)CH=CH2; 12) -ORx12, where Rx12 is -CF3 or C1-3alkyl; 13) -0H； 0 / \...basis. "!~~ 14) ' 0 , where the number is 0, 1, 2 or 3, and Rxi3 is - NH2, - NH(CHs) , -N(CH3)2, methylpyrazolyl, pyrazolyl, piperazinyl, piperidinyl, or morpholinyl; 15) , where RX14 is -CH2-, -NH-, or - 0-, and RX15 is - H, straight or branched chain Ci-5 alkyl, - NH2, -F, -Cl, -Br, or - 1; 16), where RX16 is C1-5alkyl or -CH2CH2N(CH3)2; ‘‘three’’ 1 17), ‘‘, number’’; or 18) - (C1-3 alkyl)- NH2 Ak is - CH『 , - (CH2)『 , - (CH2)3 - or , - CH『 , _ (CH2)2 _ , - •punish (CH2)3 - or ' each independently is unsubstituted or at least one of each independently is - NH2, -0H, - N02, -C1-5alkyl, - CH2NH2, -CFs, - OCF3, -CN, -F, -Cl, -Br, or -I substituted; L is a linker moiety linked to the day.

20. According to claim 19, Ri is pyridinyl, pyrazolyl, imidazolyl, phenyl, O、、,、...、••"''Heart、, s / g _,_,_,_,_,_, L,_, H '''''' I or 貢'-"""~, thin" &, and at this time, the above-mentioned R1 inside is not substituted or at least One - H is independently replaced by - NH2, -OH, -N02 or - F, At least one - H is each independently - NH2, - Ci-5alkyl, - (Ci-5alkyl) NH2, -morpholine, -piperazinyl, Substituted, Za and Zb are each independently - visible or one selected from the group consisting of the following 1) to 6), 1） -F； 366 where c is 0, 1, 2 or 3, Rx8 is piperidinyl, i 2, (the above Rb2 is - CH2 - or - NH-, and Rb3 and Rb4 are each independently - H or - NH2), RX9 is - visible; 3) - NRxwRxii, where Rxio and Rxn are both - visible; 4) Piperidinyl; 5) , where RX14 is - CH2- and RX15 is - H or - NH2; 6), where RX16 is C1-5alkyl, Ak is -CH2-, - (CH2)2 - or - (CH2)3 -, a drug-linker conjugate or a pharmaceutically acceptable salt thereof.

21. According to claim 19, 367 The day is pyridine or it, R2 is pyridine or phenyl, where phenyl is substituted with piperazine, Za and Zb are each independently - H, -F, Here, c is 2, (The above Rb2 is - NH-, and Rb3 and Rb4 are both -H), and RX9 is within; Ak is - CH2 - or - (CH2)2 -, a drug-linker conjugate or a pharmaceutically acceptable salt thereof. 【

22. In claim 19, the cytotoxic drug moiety represented by the chemical formula III is a drug-linker conjugate or a pharmaceutically acceptable salt thereof, which is any one of the compounds of Examples 1 to 43 below: 【 ​ ​ 378 [Chemical Formula IV-1] ​ ​ 【 ​ L is a drug-linker conjugate represented by the following chemical formula V, or a pharmaceutically acceptable salt thereof: [Chemical Formula V] , Ly is a single bond, or -valine-citrulline- or -glycine-glycine- phenylalanine-glycine-; Lz is a PABC (para-aminobenzyl carbamate), PAB (para-aminobenzyl), or glucuronide moiety; e is an integer from 1 to 6; h and j are each independently an integer from 1 to 3; at least two of f, g, and i are 0, and when f, g, or i are not 0, f or g is an integer from 2 to 16, and i is an integer from 16 to 30.

26. In claim 19, L is a drug-linker conjugate represented by any one of the following chemical formulae V-1 to V-10, or a pharmaceutically acceptable salt thereof: [Chemical Formula V-1] [Chemical Formula V-5] [Chemical Formula V-9] 383 【

27. ​​A drug-linker conjugate represented by any one of the following chemical formulas VI-1 to VI-11 or a pharmaceutically acceptable salt thereof: [Chemical Formula VI-1] [Chemical Formula VI-2] [Chemical Formula VI-5] 385 [Chemical Formula VI-8] 386 【

28. A urea compound represented by the following chemical formula VII, a stereoisomer thereof, and Pharmaceutically acceptable salts or hydrates or solvates thereof: [Chemical Formula VII]

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