Pyrido[2,3-b] [1,4] oxazines or tetrahydropyrido[2,3-b][1,4] oxazepines as IAP antagonists

Abstract

To provide pyrido[2,3-b] [1,4] oxazine or tetrahydropyrido[2,3-b][1,4] oxazepine derivatives as antagonists of IAPs.SOLUTION: Disclosed herein are novel 2,3-dihydro-1H-pyrido[2,3-b ][1,4] oxazine or 1,2,3,4-tetrahydropyrido[2,3-b][1,4]oxazepine derivatives used as antagonists of IAPs (Inhibitors Apoptosis Proteins), also known as Smac mimetics. Disclosed herein is the use of these antagonists for inducing or sensitizing cells to the induction of apoptotic cell death, and the use of such compounds for treating proliferative disease such as cancer.SELECTED DRAWING: None

Description

[Technical field]

[0001] As used herein, IAP (inhibitor of apoptosis proteins), also known as Smac mimetics, are The present invention relates to novel 2,3-dihydro-1H-pyrido[2, 3-b][1,4]oxazine or 1,2,3,4-tetrahydropyrido[2,3-b Derivatives of [1,4] oxazepine are disclosed herein. The use of these antagonists to induce or sensitize to induction of cell death Uses of such compounds for treating proliferative diseases, such as cancer, are also disclosed. [Background technology]

[0002] Apoptosis plays an important role in cellular development and homeostasis in higher organisms. It is a tightly regulated process for eliminating damaged or unwanted cells. (Kerr, JF, et al., Br J Cancer, 1972 , 26, 239-257). Abnormalities in the apoptotic process are known to be involved in cancer, autoimmune disorders, and cancer-related diseases. It is involved in many human diseases, including epidemics and inflammation (Nicholson, D. W., et al., Nature, 2000, 407, 810-816). In fact, resistance to apoptosis is a hallmark of cancer (Hanahan, D., et al. al., cell 2000, 100, 57-70;Hanahan, D., et al., cell, 2011, 144, 646-674).

[0003] Apoptosis is initiated by exogenous stimulation of cell death receptors or by mitochondria within the cell. May be induced by any of the endogenous stimuli produced (Elmore, S. , Toxicol Pathol, 2007, 35, 495-516). Apop -tosis inhibitory protein (IAP) is a group of extremely important negative regulators of both exogenous and endogenous apoptosis pathways . IAP was initially identified in baculoviruses and was able to inhibit apoptosis in infected cells (Birnbau m, M. J., et al., J Virol, 1994, 68, 252 1-2528). The above IAP is characterized by the presence of a baculovirus IAP repeat (BIR) domain . The BIR domain is about 70-80 amino acids long and contains a Zn-binding motif that may promote protein-protein interactions related to the function of IAP (Yang, Y.L., Cell Res, 2000, 10, 169-177 ). The human IAP family includes eight proteins, namely neuronal IAP (BIR C1), cellular IAP1 (cIAP1, BIRC2), cellular IAP2 (cIAP2, BIR C3), X chromosome-linked IAP (XIAP, BIRC4), survivin (BIRC5), ubi quitin-binding BIR domain enzyme apollon (BIRC6), melanoma IAP (ML- IAP, BIRC7), and IAP-like protein 2 (BIRC8). Among these , cIAP1, cIAP2, and XIAP play a direct role in apoptosis regulation (Salvesen, G.S., et al., Nat Rev Mol Cell Bio, 2002, 3, 401-410).

[0004] cIAP1 and cIAP2 (collectively cIAPs) inhibit the caspase-8-dependent extrinsic apoptosis pathway, such as that induced by TNF-α, via their ubiquitin ligase activity (Derakhshan, A., et al., Clin Cancer Res, 2017, 23, 1379-1387). When TNF-α binds to its receptor TNFR1, cIAPs, and tumor necrosis factor receptor type 1-associated death domain (TRADD), receptor-interacting serine / threonine kinase 1 (RIPK1) and TNF receptor-associated factor (TRAF) are recruited to form complex I, which leads to the activation of the classical nuclear factor-κB (NF-κB) pathway, which is known to promote inflammation, proliferation, and cell survival (Samuel T., et al., J Biol Chem, 2006, 281, 1080-1090; Vince J. E., et al., J Biol Chem, 2009, 284, 35906-35915; Wang C., et al., Nature, 2001, 412, 346-351).

[0005] XIAP is the only IAP protein that inhibits both extrinsic and intrinsic apoptosis pathways by directly suppressing caspase activation via its BIR domain (Deveraux Q. L., et al., Nature, 1997, 388, 300-304). The BIR2 domain of XIAP and the linker region located upstream of it associate with the IAP-binding motif (IBM) and the active site of the executioner caspases caspase-3 and -7, which are shared by both extrinsic and intrinsic apoptosis, and inhibit caspase-3 ​​​​​​​​​​​​​​​and inhibits the function of -7 (Chai J., et al., Cell, 2001 , 104, 769-780; Riedl S. J., et al., Cell , 2001, 104, 791-800). XIAP binds to procaspase-9 via its BIR3 domain and prevents the dimerization and subsequent activation of caspase-9, an important initiator caspase in the intrinsic pathway (Shiozaki E.N., et al., Mol Cell, 2003, 11, 519-527). et al., Mol Cell, 2003, 11, 519-527).

[0006] cIAP1, cIAP2, and XIAP proteins are widely expressed in various tumor types . Also, the positive expression of cIAP and XIAP is associated with high-grade cancer and poor prognosis (Che X., et al., Urol Oncol , 2012, 30, 450-456; Yang C., et al., J E xp Clin Cancer Res, 2016, 35, 158). Furthermore, downregulation or depletion of these IAPs has been shown to restore sensitivity to extrinsic or intrinsic apoptosis stimuli (Gu H., et al., Aging( Albany NY), 2018, 10, 1597-1608). In summary, targeting I AP proteins provides a potential anti-tumor strategy.

[0007] The second mitochondrial-derived caspase activator (Smac), also known as direct IAP-binding protein with low pI (DIABLO), is an endogenous antagonist of cIAP1, cIAP2, and XIAP that promotes apoptosis (Du C., et al. cIAP2, and XIAP that promotes apoptosis (Du C., et al. , Cell, 2000, 102, 33-42; Verhagen A. M., et al., Cell, 2000, 102, 43-53). Smac is normally sequestered in mitochondria and released into the cytosol upon induction of apoptosis in cells . In the cytosol, the N-terminal mitochondrial targeting sequence of Smac is cleaved, exposing the tetrapeptide (Ala-Val-Pro-Ile), which enables Smac to interact with the B IR domain of IAPs (Chai J., et al., N ature, 2000, 406, 855-862). By binding to the BIR3 domains of cIAP1 and c IAP2, Smac stimulates their E3 ubiquitin ligase activity and induces their proteasomal degradation. Loss of cIAP proteins promotes the formation of death domain-containing F as-associated protein (FADD) containing RIPK1, caspase-8, and complex II, leading to the induction of TNF-α-mediated apoptosis (Dueber E. C., et al., Science, 2011 , 334, 376-380). Dimerized Smac binds to the BIR2 and BIR3 domains of XIAP and disrupts its interaction with caspases-3, -7, and -9, which leads to caspase-dependent apoptosis (Micheau, O., et al., Cell, 2003, 114, 181-190; Chai J. , et al., Cell, 2001, 104, 769-780; Liu Z , et al., Nature, 2000, 408, 1004-1008) . . .

[0008] Smac mimetics mimic the N-terminus of Smac (Ala-Val-Pro-Ile). They are small molecules containing four amino acids. Similar to Smac, Smac mimetics bind to the BI R domain of IAPs, antagonize their functions, and promote apoptosis in cancer cells (C hai J., et al., Cell, 2001, 104, 769-780 ; Dueber E. C., et al., Science, 2011, 33 4, 376-380; Liu Z., et al., Nature, 2000, 408, 1004-1008; Verhagen A. M., et al., Cell, 2000, 102, 43-53). In summary, Smac mimetics are a novel group of cancer therapeutic drug candidates.

SUMMARY OF THE INVENTION

MEANS FOR SOLVING THE PROBLEM

[0009] In one embodiment, the present specification discloses derivatives of 2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazine or 1,2,3,4-tetrahydropyrido[2,3 -b][1,4]oxazepine of formula (I). The above embodiment includes the following aspects .

[0010] Aspect 1: Formula (I):

CHEMICAL

[0011] In some embodiments, m2 is 1, and m3 is 1, and R2 and R4 are both crosslinks that form the crosslink containing 0, 1, or 2 -CH2- moieties during the crosslinking; or or m2 is 1, and m3 is 2, and one of R2 and two R4s are both crosslinks that form the crosslink containing 0, 1, or 2 -CH2- moieties during the crosslinking; or or m2 is 2, and m3 is 1, and R2 and R4 are both crosslinks that form the crosslink containing 0, 1, or 2 -CH2- moieties during the crosslinking; or or m2 is 2, and m3 is 2, and one of two R2s and one of two R4s are both crosslinks that form the crosslink containing 0, 1, or 2 -CH2- moieties during the crosslinking. form the crosslink containing 0, 1, or 2 -CH2- moieties during the crosslinking.

[0012] In some embodiments, m2 is 1, and R 1b and R4 together form a 3 - to 6 - membered carbocyclic ring; or or m2 is 2, and R 1b and one of two R4s together form a 3 - to 6 - membered carbocyclic ring.

[0013] In some embodiments, m2 is 1, and R 1d and R4 are both crosslinks that form the crosslink containing 1 or 2 -CH2- moieties during the crosslinking; or m2 is 2, and R 1d and one of two R4s are both crosslinks that form the crosslink containing 1 or 2 -CH2- moieties during the crosslinking.

[0014] Embodiment 2: Formula (II):

Chemical formula

[0015] In some embodiments, m2 is 1, and m3 is 1, and R2 and R4 are both crosslinks, forming the above crosslink containing 0, 1, or 2 -CH2- moieties in the above crosslink; or m2 is 1, and m3 is 2, and one of R2 and two of R4 are both crosslinks, forming the above crosslink containing 0, 1, or 2 -CH2- moieties in the above crosslink; or m2 is 2, and m3 is 1, and R2 and R4 are both crosslinks, forming the above crosslink containing 0, 1, or 2 -CH2- moieties in the above crosslink; or m2 is 2, and m3 is 2, and one of two R2 and one of two R4 are both crosslinks, forming the above crosslink containing 0, 1, or 2 -CH2- moieties in the above crosslink.

[0016] In some embodiments, m2 is 1, and R 1b and R4 together form a 3 - to 6 - membered carbocyclic ring ; or m2 is 2, and R 1b and one of two R4 together form a 3 - to 6 - membered carbocyclic ring.

[0017] In some embodiments, m2 is 1, and R 1d and R4 are both crosslinks forming the above crosslink containing 1 or 2 -CH2- moieties in the above crosslink; or m2 is 2, and R 1d and one of two R4 are both crosslinks, and in the above crosslink, there is 1 ​​Or form the above crosslinking containing two -CH2- moieties.

[0018] Embodiment 3: The compound according to Embodiment 1 or 2, wherein X1 is -O-, -S-, -NH-, or -CH2-.

[0019] Embodiment 4: The compound according to Embodiment 3, wherein X1 is -CH2-.

[0020] Embodiment 5: The compound according to Embodiment 1 or 2, wherein X2 is -O-, -S-, -CH2-, or -NR a -, R a is H or -C 1~8 alkyl (methyl or ethyl). Embodiment 6: The compound according to Embodiment 5, wherein X2 is -O- or -NR

[0021] -, a and R a is hydrogen or -C 1~8 alkyl (methyl or ethyl). Embodiment 7: The compound according to Embodiment 1 or 2, wherein m1 is 0.

[0022] Embodiment 8: The compound according to Embodiment 1 or 2, wherein m2 is 1 and m3 is 0 or 1.

[0023] Embodiment 9: The compound according to Embodiment 1 or 2, wherein R2, R3, R4, and R5 are each hydrogen, methyl or ethyl, -C H2OH, -CH2OCH3, -CH2OC2H5, -CHF2, -CH2OCH(CH

[0024] 3)2, cyclopropyl, or CF3. Embodiment 10: The compound according to Embodiment 1 or 2, wherein R4 and R5 together with the same carbon atom to which they are attached form a spiro 3-5 membered carbocyclic ring.

[0025] membered carbocyclic ring. Embodiment 11: The compound according to Embodiment 1 or 2, wherein R4 and R5 together with the same carbon atom to which they are attached form a spiro 3-5

[0026] Aspect 11: [Chemical formula] is [Chemical formula] wherein R 1a , R 1b , R 1c , R 1d , R2, R3, R4, R5, and R a are the compound according to Aspect 1 or 2, as defined with respect to formula (I).

[0027] Aspect 12: [Chemical formula] is [Chemical formula] wherein R 1c , R 1d , R4, and R5 are independently hydrogen or -C 1~8 alkyl the compound according to Aspect 11.

[0028] Aspect 13: R 1c and R4 are hydrogen, and R 1d and R5 are independently hydrogen or -C 1~ 8 alkyl; preferably, R 1c and R4 are hydrogen, and R 1d and R5 are methyl the compound according to Aspect 12.

[0029] Aspect 14: R 1a , R 1b , R 1c , R 1d , R2, R3, R4, R5, and Ra is each independently hydrogen, halogen, -C alkyl (-CH3 or -C2H5, etc.) 1~8 alkyl (-CH3 or -C2H5, etc.), or oxo, provided that the above -C 1~8 alkyl is optionally substituted with at least one halogen (e.g., 1 to 3 halogens), the compound according to embodiment 11.

[0030] Embodiment 15: R5 is halogen, -C2H5, -CH3, oxo, cyclopropyl, CH F2, -CH2F, -CF3, -CH2OH, -CH2OCH3, -CH2OC2H5, -CH2OCH(CH3)2, and R4 is hydrogen or -CH3; or R5 and R4 together form a 3- to 5-membered carbon ring with the carbon atom to which they are attached; or R2 and R4 together are a bridge, forming the above bridge containing one -CH2- moiety in the above bridge and R3 and R5 are each hydrogen; or R 1b and R4 together form a 3-membered carbon ring, and R 1a and R5 are each hydrogen ; or R 1d and R4 together are a bridge, forming the above bridge containing two -CH2- moieties in the above bridge, and R 1c and R5 are each hydrogen, the compound according to embodiment 14.

[0031] Embodiment 16:

Chemical formula

Chemical formula

Chemical formula

[0032] Embodiment 17: [Chemical formula] is [Chemical formula] The compound according to embodiment 1 or 2.

[0033] Embodiment 18: The compound according to embodiment 17, wherein R7 is halogen.

[0034] Embodiment 19: The compound according to embodiment 18, wherein R7 is F.

[0035] Embodiment 20: R6 is hydrogen, -CN, halogen, morpholine, -CONR a R b , -OR a , -NR a R b , -C 3~5 cycloalkyl, phenyl, or -C 1~8 alkyl wherein the -C 1~8 alkyl or -C 3~5 cycloalkyl is optionally substituted with at least one R d ; R a and R b are each hydrogen, -C 3~6 cycloalkyl, or -C 1~8 alkyl ; R d is, in each instance of its presence, independently, -CH2F, -CHF2, -CF3, - F, -Cl, -Br, -I, -OH, -NH2, -SH, -CN, -CONH2, -H, -CH3, -C2H5, -C3H7, -CH2OH, -OCH3, -OC2H5, morpho lino,

Chem.

[0036] Embodiment 21: R6 is -CONR a R b , -NR a R b , NR a CO-R b , -NR a - SO2-R b , or -OR a ; R a and R b are each hydrogen, -C 3~6 cycloalkyl, -C 1~8 alkyl, 5 or 6-membered heterocyclyl, aryl, or heteroaryl, provided that the above -C 1~8 alkyl, -C 3~6 cycloalkyl, 5- or 6-membered heterocyclyl, aryl , or heteroaryl is optionally substituted with 1 to 3 R d ; or R a and R b together with the atom(s) to which they are attached form a 3- to 8-membered ring, provided that the above ring contains 0, 1, or 2 additional heteroatoms independently selected from nitrogen, oxygen, or optionally oxidized sulfur as ring members, and the above ring is optionally substituted with 1 to 3 R d ; R d is, in each instance of its presence, independently hydrogen, halogen, -C 1~8 alkyl, -C 2~8Alkenyl, -C 2~8 Alkynyl, -C 3~8 Cycloalkyl, heterocycl yl, aryl, heteroaryl, oxo, -CN, -NO2, -OR e , -SR a , - SO2R e , -SO2NR e R f , -COR e , -CO2R e , -CONR e R f , -C (=NR e )NR f R g , -NR e R f , -NR e COR f , -NR e CONR f R g , -NR e CO2R f , -NR e SONR f R g , -NR e SO2NR f R g , or -N R e SO2R f wherein each of the above -C 1~8 alkyl, -C 2~8 alkenyl, -C2 ~8 alkynyl, -C 3~8 cycloalkyl, heterocyclyl, aryl, or hetero aryl is optionally substituted with at least one substituent selected from halogen, -C 1~8 alkyl, -OR h , -NR h R i , cycloalkyl, heterocyclyl, aryl, or heteroaryl; ; R e , R f , Rg , R h , and R i are each independently hydrogen, -C 1~8 alkyl, C 1~8 alkoxy-C 1~8 alkyl-, -C 2~8 alkenyl, -C 2~8 alkynyl , cycloalkyl, heterocyclyl, aryl, aryl-C 1~8 alkyl-, or heteroaryl, the compound according to embodiment 1 or 2.

[0037] Embodiment 22: R6 is -CONR a R b or NR a CO-R b and R a and R b are each hydrogen, -C 1~3 alkyl, -C 3~6 cycloalkyl, ph enyl, 5- or 6-membered heterocyclyl (azetidine, pyrrolidine, piperidine, oxa zolidine, isoxazolidine, oxazinane, morpholine, piperazine, tetrahydro pyran, or tetrahydrofuran, etc.), provided that the above -C 1~3 alkyl, -C 3~6 cycloalkyl, phenyl, 5- or 6-membered heterocyclyl (azetidine, pyrrol idine, piperidine, oxazolidine, isoxazolidine, oxazinane, morpholine, piperazine, tetrahydropyran, or tetrahydrofuran, etc.) are optionally substituted with at least one substituent selected from halogen, -C 1~3 alkyl, morpholine, OR e , or -NR e R f ; and R R e and Rf is independently hydrogen, halogen, -C 1~8 alkyl, or C 1~8 alko xy-C 1~8 alkyl-; a compound according to embodiment 21.

[0038] Embodiment 23: R6 is -CONR a R b ; and R R a and R b together with the nitrogen atom to which they are attached form a 4- to 6-membered ring, provided that the said ring contains 0, 1 or optionally up to 2 further heteroatoms selected independently from nitrogen, oxygen or optionally oxidized sulfur as ring members; and the said ring is optionally substituted with 1 to 3 R d ; R is independently in each of its occurrences halogen, -C d alkyl, or 1~3 -OR ; R e is independently hydrogen, halogen, or -C R e alkyl; a compound according to embodiment 21. 1~8

[0039] Embodiment 24: R6 is

Chem.

Chem.

[0040] Embodiment 25: R6 is hydrogen, halogen, -CN, -CONH2,

Chem.

[0041] Embodiment 26: R 12 and R 13 are each independently hydrogen or -C 1~8 alkyl the compound according to embodiment 1 or 2.

[0042] Embodiment 27: R 12 is hydrogen or -CH3; R 13 is hydrogen or -CH3, the compound according to embodiment 26.

[0043] Embodiment 28: R 10 and R 11 are each independently hydrogen, -C 1~8 alkyl, -C 3~ 8-cycloalkyl, -C 5~6 aryl, 5- to 6-membered heteroaryl, or -CONR a R b wherein the above -C 1~8 alkyl, -C 3~8 cycloalkyl, -C 5~6 a ryl, or 5- to 6-membered heteroaryl is optionally substituted with 1 to 3 R d ; ; R a and R b are each independently hydrogen or -C 1~8 alkyl; or (R a and R b ) together with the nitrogen atom to which they are attached form a 3- to 6-membered ring, provided that the above ring contains 0 or 1 additional oxygen atom; the above ring is optionally substituted with one R d ; ; R dis, independently in each of its occurrences, -C 1~8 alkyl, halogen, aryl or -OR e wherein R e is independently hydrogen or -C 1~8 alkyl phenyl ethyl, benzyl or phenyl, a compound according to embodiment 1 or 2.

[0044] Embodiment 29: R 10 is hydrogen; R 11 is -CONR a R b phenyl, benzyl, pyridinyl or furyl wherein each of said phenyl, benzyl, pyridinyl or furyl is optionally substituted with 1 to 3 R d selected from methyl, methoxy or halogen; R a and R b are each independently hydrogen or methyl or (R a and R b ) together with the nitrogen atom to which they are attached form a 5- or 6-membered ring wherein said ring contains 0 or 1 additional oxygen atom, a compound according to embodiment 28.

[0045] Embodiment 30: R 10 is hydrogen; R 11 is hydrogen, methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, tert t-butyl, 1-methylpropyl, 1,1-dimethylethyl, 1-pentyl, 2-pentyl 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1 -butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2- Methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-meth yl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3, 3-dimethyl-2-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohe xyl, vinyl, -CH2OH, -CH2CH2OH, -CH2CH2OCH3, -CH 2OCH3, -CH2OCH(CH3)2, -CH2OC2H5, -CH2F, -CHF 2, -CF3, -H, -F, -Cl, -Br, -I, -OH, -NH2, -SH, -CO NH2, -CONHCH3, -CON(CH3)2,

Chem.

[0046] Embodiment 31: R 10 is hydrogen; R 11 is hydrogen, methyl, ethyl, 2-propyl, -CF3, -CHF2, -CH2C H2OCH3, -CH2OH, -CH2OCH3, -CH2OCH(CH3)2, -CH 2OC2H5, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -C ONH2, -CONHCH3, -CON(CH3)2,

Chem.

[0047] Embodiment 32: R 10 and R 11 are different, R 10 and R 11 are attached to the carbon atom is in the (S) configuration, the compound according to any one of embodiments 1 to 31.

[0048] Embodiment 33: R 10 and R 11 together form a 3-membered carbon ring, the compound according to embodiment 1 or 2 of the compound.

[0049] Embodiment 34:

Chemical formula

[0050] Embodiment 35: R 14 and R 15 are each independently hydrogen or -C 1~8 alkyl (methyl or ethyl) selected from the compound according to embodiment 1 or 2.

[0051] Embodiment 36:

Table 1-1

Table 1-2

Table 1-3

Table 1-4

Table 1-5

Table 1-6

Table 1-7

Table 1-8

Table 1-9

Table 1-10

Table 1-11

[0052] In some embodiments, as used herein, the compounds disclosed herein, or pharmaceutically acceptable salts thereof, or stereoisomers, tautomers, or prodrugs thereof, and A pharmaceutical composition comprising at least one pharmaceutically acceptable carrier or excipient is disclosed. .

[0053] In some embodiments, as used herein, a method of treating a disease responsive to inhibition of cIAP, the method comprising administering to a subject in need thereof a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a stereoisomer, tautomer, or prodrug thereof. is disclosed.

[0054] In some embodiments, a method of treating cancer modulated by cIAPs, the method comprising administering to a subject in need thereof a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a stereoisomer, tautomer, or prodrug thereof. is disclosed. DETAILED DESCRIPTION OF THE INVENTION

[0055] The following terms have the meanings set forth throughout this specification.

[0056] As used herein, including the appended claims, the singular forms of the words "a," "an," and "the" include the corresponding plural referents unless the context clearly dictates otherwise.

[0057] As used herein, unless the context clearly dictates otherwise, the term "or" is used to mean "and / or" and is synonymous therewith.

[0058] The term "alkyl" refers to a straight-chain, branched-chain, or cyclic hydrocarbon radical having from 1 to 18 carbon atoms, such as from 1 to 12 carbon atoms, more preferably from 1 to 10 carbon atoms, even more preferably from 1 to 8 carbon atoms, or from 1 to 6 carbon atoms, or from 1 to 4 carbon atoms, etc. -chain, branched-chain, or cyclic hydrocarbon radical having from 1 to 18 carbon atoms, such as from 1 to 12 carbon atoms, more preferably from 1 to 10 carbon atoms, even more preferably from 1 to 8 carbon atoms, or from 1 to 6 carbon atoms, or from 1 to 4 carbon atoms, etc.​​​​​​​ Refers to a hydrocarbon group selected from linear and branched saturated hydrocarbon groups. An alkyl group containing 1 to 6 carbon atoms (i.e., C alkyl) examples include methyl, ethyl, 1 1~6 -propyl i.e., n-propyl ("n-Pr"), 2-propyl i.e., isopropyl ("i-Pr"), 1-butyl i.e., n-butyl ("n-Bu"), 2-methyl-1- propyl i.e., isobutyl ("i-Bu"), 1-methylpropyl i.e., sec-but yl ("sec-Bu"), 1,1-dimethylethyl i.e., t-butyl ("t-Bu" ), 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl -2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2- hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4- methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3 -dimethyl-2-butyl, and 3,3-dimethyl-2-butyl groups, but are not limited to these.

[0059] The term "cycloalkyl" refers to a saturated cyclic hydrocarbon group selected from monocyclic groups and polycyclic (e.g., bicyclic and tricyclic) groups including fused, bridged, or spirocycloalkyl groups.

[0060] The term "aryl", used alone or in combination with other terms, refers to a group selected from the following. · 5- and 6-membered carbocyclic aromatic rings, e.g., phenyl, · 7- to 12-membered bicyclic ring systems in which at least one ring is carbocyclic and aromatic, e.g., naphthyl and indanyl, and At least one ring is carbocyclic and aromatic, e.g., fluorenyl, Tricyclic ring systems such as 15-membered tricyclic ring systems.

[0061] The terms "aromatic hydrocarbon ring" and "aryl" are used interchangeably throughout this disclosure. In some embodiments, the monocyclic or bicyclic aromatic hydrocarbon ring is 10 to 10 ring-forming carbon atoms (i.e., C 5~10 Aryl). Monocyclic or bicyclic Examples of cyclic aromatic hydrocarbon rings are phenyl, naphth-1-yl, and naphth-2-yl. Examples of the aryl group include, but are not limited to, anthracenyl, and phenanthrenyl. In some embodiments, the aromatic hydrocarbon ring is a naphthalene ring (naphthalene-1-yl). or naphth-2-yl) or a phenyl ring. The aromatic hydrocarbon ring is a phenyl ring.

[0062] The term "aryl-alkyl-" refers to any of the above defined groups further substituted with an aryl group. An example of an aryl-alkyl group is aryl-C 1~8 a alkyl, for example, phenylethyl, or phenylmethyl (benzyl).

[0063] The term "heteroaryl" refers to a group selected from: At least one heteroatom selected from nitrogen (N), sulfur (S), and oxygen (O). Some of the members may be, for example, 1 to 4, or in some embodiments, 1 to 3. In some embodiments, the ring contains 1-2 heteroatoms and the remaining ring atoms are carbon. is a seven-membered aromatic monocyclic ring, At least one heteroatom selected from N, O, and S, for example, 1 to 4, or, in some embodiments, 1 to 3, and in some embodiments 1 to 2 heteroatoms, with the remaining ring atoms being carbon, a 7- to 12-membered bicyclic ring, wherein at least one ring is aromatic and at least one heteroatom is present in the aromatic ring, said bicyclic ring, and · at least one heteroatom selected from N, O, and S, for example, 1 to 4, and also or, in some embodiments, 1 to 3, or in other embodiments 1 or 2 heteroatoms, with the remaining ring atoms being carbon, an 11- to 14-membered tricyclic ring, wherein at least one ring is aromatic and at least one heteroatom is present in the aromatic ring, said tricyclic ring.

[0064] When the total number of S and O atoms in the heteroaryl group exceeds 1, those hetero atoms are not adjacent to each other. In some embodiments, the total number of S atoms and O atoms in the heteroaryl group is 2 or less. In some embodiments, the total number of S atoms and O atoms in the aromatic heterocyclic ring is 1 or less. When the heteroaryl group contains two or more heteroatom-containing rings, the heteroatoms may be the same or different. The nitrogen atoms in the ring(s) of the heteroaryl group may be oxidized to form N-oxides. As used herein, the term "C-bonded heteroaryl" means that the heteroaryl group is bonded to the core molecule by a bond from a C atom of the heteroaryl ring. .

[0065] The terms "aromatic heterocyclic" and "heteroaryl" are synonymous throughout the disclosure of this is used. In some embodiments, the monocyclic or bicyclic aromatic heterocycle has 5, 6 , 7, 8, 9, or 10 ring members and contains 1, 2, 3, or 4 heteroatom ring members independently selected from nitrogen (N), sulfur (S), and oxygen (O ), and the remaining ring members are carbon. In some embodiments, the monocyclic or bicyclic aromatic heterocycle is a monocyclic or bicyclic ring containing 1 or 2 heteroatom ring members independently selected from nitrogen (N), sulfur (S), and oxygen (O ). In some embodiments, the monocyclic or bicyclic aromatic heterocycle is a 5- to 6-membered heteroaryl ring, the ring is monocyclic, and has 1 or 2 heteroatoms independently selected from nitrogen (N), sulfur (S), and oxygen (O ). In some embodiments, the monocyclic or bicyclic aromatic heterocycle is an 8- to 10-membered heteroaryl ring, the ring is bicyclic, and has 1 or 2 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen .

[0066] "Heterocyclyl", "heterocycle" or "heterocyclic" are synonymous and include one or more heteroatoms selected from nitrogen, oxygen, or optionally oxidized sulfur as ring members, with the remaining ring members being carbon, and include monocyclic rings, fused rings, bridged rings, and spiro rings, i.e., non-aromatic heterocyclyl groups including monocyclic heterocyclyl, bridged heterocyclyl, spiro heterocyclyl, and fused heterocyclic groups. As used herein, the term "optionally oxidized sulfur" refers to S, SO, or SO2.

[0067] The compounds disclosed herein may contain asymmetric centers and, therefore, may exist as enantiomers. "Enantiomers" refers to two stereoisomers of a compound that are non-superimposable mirror images of each other. When the compounds disclosed herein have two or more asymmetric centers, they may exist further as diastereomers. Enantiomers and diastereomers belong to a broader group of stereoisomers. All such possible stereoisomers are intended to be included, including substantially pure separated enantiomers, their racemic mixtures, and mixtures of diastereomers. All stereoisomers of the compounds disclosed herein and / or their pharmaceutically acceptable salts are intended to be included. Unless otherwise specifically mentioned, a reference to one isomer applies to any of the possible isomers. Whenever the isomeric composition is not specified, all possible isomers are included. The term "substantially pure" as used herein means that the desired stereoisomer contains no other stereoisomer(s) in excess of 35% by weight, for example, in excess of 30% by weight, further for example, in excess of 25% by weight, still further for example, in excess of 20% by weight. In some embodiments, the term "substantially pure" means that the desired stereoisomer contains no other stereoisomer(s) in excess of 10% by weight, for example, in excess of 5% by weight, for example, in excess of 1% by weight. If the compounds disclosed herein contain an olefinic double bond, unless otherwise specified, such double bond is meant to include both E and Z geometric isomers.

[0068]

[0069]

[0070] ​​​​​​​​​​​​If the compounds disclosed in this specification contain a disubstituted cyclohexyl group or a cyclobutyl group, the substituents present on the cyclohexyl ring or cyclobutyl ring may adopt cis and trans conformations. The cis conformation means that both substituents are present on the upper side of the arrangement of the two substituents on the carbon, while trans means that the two substituents are present on the opposite sides.

[0071] It may be advantageous to separate the multiple reaction products from each other and / or from the starting materials. The desired product of each step or series of steps is separated and / or purified to the desired degree of homogeneity by techniques common in the art (hereinafter referred to as "separated"). Generally, such separation includes liquid-liquid extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography includes, for example, reverse and normal phase chromatography; size exclusion chromatography; ion exchange chromatography; high, medium, and low pressure liquid chromatography methods and apparatuses; small scale analytical chromatography; simulated moving bed ("SMB") chromatography and preparative thin or thick layer chromatography; and small scale thin layer chromatography and flash chromatography techniques, and may include any number of methods. One of ordinary skill in the art will apply the technique most likely to achieve the desired separation.

[0072] "Diastereomer" refers to stereoisomers of a compound that have two or more chiral centers but are not mirror images of each other. A mixture of diastereomers can be separated into the individual diastereomers of the mixture by methods well known to those skilled in the art, such as chromatography and / or fractional crystallization. ​ Based on the physicochemical differences of the mers, they can be separated into the above individual diastereomers. Enantiomers can be separated by reacting the enantiomer mixture with a suitable optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher acid chloride) to convert it into a diastereomer mixture, separating the diastereomers, and converting the individual diastereomers into the corresponding pure enantiomers (e.g., by hydrolysis). Enantiomers can also be separated by using a chiral HPLC column.

[0073] The term "pharmaceutically acceptable salt" refers to a salt that, within the scope of sound medical judgment, is suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and that has a reasonable benefit / risk ratio. Pharmaceutically acceptable salts can be prepared in situ or separately during the final isolation and purification of the compounds disclosed herein by reacting the free base functional group with a suitable organic acid or by reacting an acidic group with a suitable base.

[0074] When an acid is added and the compounds disclosed herein are obtained as acid addition salts, the free base can be obtained by basifying the solution of the acid salt. Conversely, when the product is a free base, the free base can be dissolved in a suitable organic solvent and / or water and the solution can be treated with an acid according to conventional procedures for producing acid addition salts, such as pharmaceutically acceptable addition salts. Those skilled in the art will recognize various synthetic methodologies that can be used to produce non-toxic pharmaceutically acceptable addition salts without undue experimentation. ​

[0075] As defined herein, a "pharmaceutically acceptable salt thereof" is a compound represented by the formula (I): Salts of compounds and salts of stereoisomers of compounds of formula (I), e.g. salts of enantiomers and / or or diastereomeric salts thereof.

[0076] As used herein, the terms "administration," "administering," "treat," and "treating" refer to , animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, Administering exogenous drugs, therapeutic agents, or other substances to an animal, human, subject, cell, tissue, organ, or biological fluid. Treatment of a cell means contacting the cell with a reagent, such as a diagnostic agent, or a composition. The term "injection" also includes contacting a fluid with said cells. "Administering" and "treating" also refer to in vitro and ex vivo treatments, e.g., administering to cells, reagents, It also refers to treatment with a diagnostic agent, a binding compound, or with another cell. The term "subject" refers to any organism, preferably an animal, more preferably a mammal (e.g., a rat). , mouse, dog, cat, and rabbit), and most preferably human.

[0077] The term "effective amount" or "therapeutically effective amount" refers to an amount that reduces the severity of a disease or clinical symptoms of a disease or disorder. When administered to a subject to treat at least one of the above diseases, disorders, or conditions, The above-mentioned " A "therapeutically effective amount" refers to a therapeutically effective amount of the compound, the disease, disorder, and / or the symptoms of the disease or disorder. the condition, said disease, disorder, and / or symptoms of said disease or disorder, the age of the subject receiving treatment The dosage may vary depending on the age and / or weight of the subject being treated. The appropriate amount in an example can be apparent to those skilled in the art or can be determined by conventional experiments. In some embodiments, a "therapeutically effective amount" is at least one compound disclosed herein and / or at least one of its stereoisomers, and / or at least one pharmaceutically acceptable salt thereof that is effective for "treating" a disease or disorder in a subject as defined above. In the case of combination therapy, a "therapeutically effective amount" refers to the total amount of the combination objects for the effective treatment of a disease, disorder, or illness. The pharmaceutical composition containing the compound disclosed herein can be administered to a subject in need thereof via oral administration, inhalation, rectal administration, parenteral administration, or topical administration. In the case of oral administration, the pharmaceutical composition may be a conventional solid preparation, such as tablets, powders, granules, capsules, etc., a liquid preparation, such as a suspension in water or oil, or other liquid preparations such as syrups, solutions, suspensions, etc. In the case of parenteral administration, the pharmaceutical composition may be a solution, an aqueous solution, an oil suspension concentrate, a lyophilized powder, etc.

[0078] The above preparations of the pharmaceutical composition are preferably selected from tablets, coated tablets, capsules, suppositories, nasal sprays, or injections, more preferably tablets or capsules. The pharmaceutical composition may be a single unit dose of an exact dosage. Additionally, the pharmaceutical composition may further contain additional active ingredients. All preparations of the pharmaceutical composition disclosed herein can be manufactured by conventional methods in the pharmaceutical field. For example, the active ingredient is mixed with one or more excipients and then the desired preparation is manufactured.

[0079] It can be done. "Pharmaceutically acceptable excipients" refer to conventional pharmaceutical carriers suitable for the desired pharmaceutical formulation, such as diluents, water, vehicles such as various organic solvents, starches, sucrose, and other fillers; binders such as cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone ( PVP); wetting agents such as glycerin; disintegrants such as agar, calcium carbonate, and sodium bicarbonate ; absorption promoters such as quaternary ammonium compounds; surfactants such as hexadecanol ; absorption carriers such as kaolin and soap clay; lubricants such as talc, calcium stearate, magnesium stearate, and polyethylene glycol . When added, the above pharmaceutical composition further contains other pharmaceutically acceptable excipients such as dispersants, stabilizers, thickeners, complexing agents, buffers, penetration enhancers, polymers, aromatic compounds, sweeteners, and coloring agents.

[0080] The term "disease" refers to any disease, discomfort, illness, symptom, or sign, and can be synonymous with the terms "disorder" or "ailment".

[0081] Throughout this specification and the subsequent aspects, unless the context otherwise requires, the terms "comprise" , "comprises", and variations such as "comp rising" are intended to identify the presence of the subsequent features, but do not exclude the presence or addition of one or more other features. As used herein, the term "comprising" can be equivalently expressed as the terms "containing" , "including", and optionally "having".

[0082] Throughout this specification and the following aspects, the term "C" n~m ", where n and m are integers, represents a range including endpoints that indicates the number of carbon atoms. Examples include C 1~8 , C 1~6 , etc. can be mentioned.

[0083] Unless otherwise specifically defined elsewhere in this written document, all other technical terms and scientific terms used in this specification have the meaning generally understood by those skilled in the technical field to which the present invention belongs.

[0084] General synthesis The compounds (including their salts) disclosed in this specification can be produced using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes. The reactions for producing the compounds disclosed in this specification can be carried out in a suitable solvent that can be readily selected by one skilled in the art of organic synthesis. Suitable solvents can be substantially non-reactive with the starting materials, intermediates, or

[0085] products at the temperature at which the reaction is carried out, for example, in the range from the boiling point of the solvent. A given reaction can be carried out in a single solvent or a mixture of multiple solvents. The selection of suitable protecting groups can be readily determined by one skilled in the art. The reaction can be monitored by any and appropriate methods known in the art, such as NMR, UV, HPLC, LC-MS, and TLC. The compounds can be purified by various methods including HPLC and normal-phase silica chromatography.

[0086]

[0087]

[0088] ​​​​​​Scheme I [ka] Compounds of formula (I) can be prepared as shown in Scheme I. Nucleophilic substitution between the aryl and compound (i) gives compound (ii), which is then catalyzed by iron. Compound (iii) is obtained by reductive cyclization mediated by borane. Compound (iv) was obtained by reduction mediated by 3-methyl-2-propanediol. (vi) was then obtained by Negishi or Suzuki coupling to give compound ( Compound (vii) was obtained. Compound (ix) was obtained by deprotection and subsequent acylation. Compound (ix) was reacted with key intermediate (x) to give compound (xi). Thus, compound (xii) represented by formula (I) was obtained.

[0089] Scheme II [ka] Compounds of formula (I) can also be synthesized as shown in Scheme II. Nucleophilic substitution between i) and a Boc-protected amino alcohol gave compound (ii). to give compound (iii) by Cu or Pd mediated C-N coupling, Subsequent Negishi or Suzuki coupling gives compound (iv). Compound (v) was obtained by protection. Compound (v) was reacted with 2-chloroacetyl chloride to give Compound (vi) was obtained by reacting compound (vi) with key intermediate (vii) to obtain compound (vi). (viii) was obtained. Finally, the compound (ix) represented by formula (I) was obtained by deprotection.

[0090] Abbreviations JPEG2025090788000032.jpg226161

Example

[0091] Example 1: 7-(4-Fluorobenzyl)-1-(2-((2R,5R)-5-methyl- 2-(((R)-3-Methylmorpholino)methyl)piperazin-1-yl)acetyl)- 2,3-Dihydro-1H-pyrido[2,3-b][1,4]oxazine-6-carbonitrile rilginate (Compound 1)

Chem.

Chem.

[0092] Step 2: 7-Bromo-1H-pyrido[2,3-b][1,4]oxazin-2(3H )-one [Chemical formula] Ethyl 2-((5-bromo-3-nitropyridin-2-yl)oxy)acetate (13 g, 42.6 mmol) in AcOH (300 mL) was slowly added with Fe powder (13 g, 23 2 mmol) at 70 °C, and the resulting mixture was stirred at 70 °C for 1 hour. AcOH was removed in vacuo, and the resulting solid was washed with MeOH (200 mL) and filtered. This operation was repeated 5 times. The MeOH phase was concentrated in vacuo to obtain the crude product (9.6 g, yield 98%), which was used directly in the next step. LC-MS (M+H) + = 228 .9, 230.9.

[0093] Step 3: 7-Bromo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine [Chemical formula] 7-Bromo-1H-pyrido[2,3-b][1,4]oxazin-2(3H)-one ( 9 g, 39.3 mmol) in anhydrous THF (150 mL) was added dropwise with BH3 (98.3 mL, 1N THF solution) at room temperature, and the resulting solution was stirred at 60 °C for 1 hour. This reaction mixture was cooled to room temperature, quenched with MeOH (10 mL), adjusted to pH 1-2 with the addition of 1N HCl, and stirred at 60 °C for 1 h. This reaction mixture was cooled to room temperature and water (100 mL) was added, and the pH was adjusted to 8-9. This mixture was extracted with EtOAc (100 mL × 3), the combined organic layers were dried over Na2SO4, filtered, and in vacuo ​​It was distilled off to obtain a crude product (7.46 g, yield 88%), which was used directly in the next step. LC-MS (M+H) + = 215.1, 217.1.

[0094] Step 4: 7-Bromo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine tert-Butyl 7-bromo-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate

Chemical formula

[0095] Step 5: 7-Bromo-1-(tert-butoxycarbonyl)-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazine 5-oxide

Chemical formula

[0096] Step 6: 7-Bromo-6-cyano-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazine-1-carboxylic acid tert-butyl [Chemical formula] To a solution of 7-bromo-1-(tert-butoxycarbonyl)-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazine 5-oxide (2.9 g, 8.8 mmol) in MeCN (100 mL) was added TMSCN (13.04 g, 131.4 m mol) at room temperature under N2, and the resulting solution was stirred at 80 °C for 12 h. The reaction mixture was concentrated in vacuo and the residue was purified by silica gel chromatography (DCM:MeOH = 50:1) to give the above product (715 mg, yield 24%). LC-MS (M+H) = 340.2, 342.2. +

[0097] Step 7: 6-Cyano-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl [Chem.] 7-Bromo-6-cyano-2,3-dihydro-1H-pyrido[2,3-b][1,4] tert-Butyl 7-bromo-6-cyano-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate (715 mg, 2.1 mmol) and bis (tri-tert-butylphosphine)palladium (54 mg, 0.11 mmol) in anhydrous THF (100 mL) was added dropwise to (4-fluorobenzyl)zinc chloride (II) (8.4 m L, 4.2 mmol, 0.5 M THF solution), and the resulting solution was stirred at 60 °C for 2 hours . The reaction mixture was quenched with MeOH (5 mL), concentrated in vacuo, and the residue was purified by silica gel chromatography (PE:EtOAc = 5:1) to give the above product (470 mg, 61% yield). LC-MS (M+H) = 370.4. +

[0098] Step 8: 7-(4-Fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3 -b][1,4]oxazine-6-carbonitrile [Chem.] 6-Cyano-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2, 3-b][1,4]oxazine-1-carboxylic acid tert-butyl (360 mg, 0.9 8 mmol) in DCM (10 mL) was added to a 1,4-dioxane solution of HCl (4 N, 2 .5 mL, 10 mmol), and the resulting solution was stirred at room temperature for 12 hours. This solution was concentrated in vacuo. The mixture was neutralized with aqueous NaHCO3 and extracted with EtOAc . The combined organic layers were dried, filtered, and concentrated to give the title compound (260 mg, yield​ Yield 99%). LC-MS (M+H) + = 270.4

[0099] Step 9: 1-(2-Chloroacetyl)-7-(4-fluorobenzyl)-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazine-6-carbonitrile

Chem.

[0100] Step 10: Methyl ((benzyloxy)carbonyl)-L-seryl-D-alanine (methyl ((benzyloxy)carbonyl)-L-seryl-D -alaninate)

Chem.

[0101] Step 11: (3S,6R)-3-(hydroxymethyl)-6-methylpiperazine-2 ,5-dione [ka] Under nitrogen, methyl((benzyloxy)carbonyl)-L-seryl-D-alaninate (10 g, 30.8 mmol), 10% Pd / C (500 mg), MeOH (100 C. (65 mL) and cyclohexene (65 mL) were added. The mixture was heated to reflux. The hot reaction mixture was filtered through Celite, and the cake was diluted with hot MeOH. The combined filtrate was concentrated. The resulting solid was slurried in 2-butanone. The mixture was cooled to room temperature and petroleum ether was slowly added over 10 minutes. After stirring for 30 minutes, the solids were The cake was filtered and further washed with a mixture of PE / 2-butanone (v / v=2 / 1). The product was obtained after filtration and drying (2.5 g, 51% yield). LC-MS (M+H) + = 159.1.

[0102] Step 12: ((2R,5R)-5-Methylpiperazin-2-yl)methanol hydrochloride [ka] (3S,6R)-3-(Hydroxymethyl)-6-methylpiperazine-2,5-dione (2.5 g, 15.8 mmol) was added to a THF solution of borane (1 M, 126 mL, 126. 4 mmol), and the mixture was heated at 70 °C for 18 h. The solution was cooled in ice, and then M eOH (30 mL) was added dropwise, followed by 5N HCl (8 mL). The mixture was heated at 70 °C for 2 h and then cooled to room temperature. The resulting solid was filtered, washed with TH F, and dried to obtain the above product (2.0 g, 77% yield). LC-MS (M+ H) = 131.1. +

[0103] Step 13: (2R,5R)-5-(Hydroxymethyl)-2-methylpiperazine-1 -carboxylic acid tert-butyl

Chemical Structure

[0104] Step 14: (2R,5R)-4-Benzyl-5-(hydroxymethyl)-2-methyl tert-butyl piperazine-1-carboxylate

Chem.

[0105] Step 15: (2R,5R)-4-Benzyl-5-(chloromethyl)-2-methylpipe radine-1-carboxylic acid tert-butyl

Chem.

[0106] Step 16: (2R,5S)-4-Benzyl-2-methyl-5-(((R)-3-methyl morpholino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0107] Step 17: (2R,5S)-2-Methyl-5-(((R)-3-methylmorpholino) methyl)piperazine-1-carboxylic acid tert-butyl [Chemical formula] (2R,5S)-4-Benzyl-2-methyl-5-(((R)-3-methylmorpholino )methyl)piperazine-1-carboxylic acid tert-butyl (1.3 g, 3.2 mmol) To a solution of this in EtOH (20 mL), Pd / C (130 mg) and a few drops of AcOH were carefully added. The mixture was stirred at ambient temperature for 3 hours under a H2 atmosphere. The mixture was concentrated and extracted with DCM. The combined organic layers were washed with saturated NaHCO3 solution, dried, filtered, and concentrated. The resulting title compound (1.0 g) was used in the next step without purification LC-MS (M+H)+ = 314.2.

[0108] Step 18: (2R,5S)-4-(2-(6-Cyano-7-(4-fluorobenzyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl) -2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl) piperazine-1-carboxylic acid tert-butyl [Chemical formula] 1-(2-Chloroacetyl)-7-(4-fluorobenzyl)-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazine-6-carbonitrile (68 mg, 0. 196 mmol), (2R,5S)-2-methyl-5-(((R)-3-methylmorpho olino)methyl)piperazine-1-carboxylic acid tert-butyl (62 mg, 0.196 mm ol), KI (65 mg, 0.392 mmol), and K2CO3 (81 mg, 0.58 A solution of MeCN (10 mL) of 8 mmol was stirred at room temperature for 2 h. The reaction mixture was concentrated in vacuo to remove the solvent, and the residue was washed with H2O (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were concentrated and purified by preparative TLC (DCM:MeOH = 20:1) to give the title compound (80 mg, yield 65.5%). LC-MS (M+H) = 623.7. In vacuo, the solvent was removed, and the residue was washed with H2O (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were concentrated and purified by preparative TLC (DCM:MeOH = 20:1) to give the title compound (80 mg, yield 65.5%). LC-MS (M+H) = 623.7. MS (M+H) + = 623.7.

[0109] Step 19: 7-(4-Fluorobenzyl)-1-(2-((2R,5R)-5-methyl -2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl)acetyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-6-carbo nitrile formate (Compound 1) (2R,5S)-4-(2-(6-Cyano-7-(4-fluorobenzyl)-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxo ethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine- 1-carboxylic acid tert-butyl (80 mg, 0.128 mmol) in DCM (10 mL ) was added to a 1,4-dioxane solution of hydrochloric acid (4 N, 2.5 mL), and the resulting solution was stirred at room temperature for 4 h. The solution was concentrated in vacuo, dissolved in MeOH (10 mL), adjusted to pH 8-9 by the addition of an aqueous NaHCO3 solution, and then concentrated in vacuo to remove the solvent and the residue was purified by preparative HPLC (gradient eluent: 10% - 25% CH3CN / H2O (each containing 0.1% formic acid (FA))) to give Compound 1 as the formate (30 mg, yield 41%). (30 mg, yield 41%). 1 1H NMR (400 MHz, DMSO-d6) δ 1 1.31 - 9.78 (m, 1H), 8.63 (s,1H), 8.26 (s, 2 H), 7.31 - 7.22 (m, 2H), 7.22- 7.11 (m, 2H), 4.53 - 4.34(m, 2H), 4.17 -3.79 (m, 6H), 3.5 2 - 3.31 (m, 2H), 3.26 - 2.53(m, 10H), 2.23 - 2.09 (m, 1H), 2.04 - 1.92(s, 1H), 1.91 - 1 .78 (m, 1H), 1.08 - 1.01 (m,3H), 0.90 - 0.82 (m, 3H). LC-MS(M+H) + = 523.5。

[0110] Example 2: 1-(7-(4-Fluorobenzyl)-2,3-dihydro-1H-pyrido[2 ,3-b][1,4]oxazin-1-yl)-2-((2R,5R)-5-methyl-2 -(((R)-3-Methylmorpholino)methyl)piperazin-1-yl)ethan-1-one formate (Compound 2)

Chem.

Chem.

[0111] Step 2: 7-(4-Fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3 -b][1,4]oxazine

Chemical formula

[0112] Step 3: 2-Chloro-1-(7-(4-fluorobenzyl)-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one

Chemical formula

[0113] Step 4: (2R,5S)-4-(2-(7-(4-fluorobenzyl)-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoeth yl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazin-1 tert-Butyl -carboxylate [Chem.] The title compound (80 mg) of Step 4 was prepared in the same manner as described in Step 18 of Example 1 from 2-chloro-1-(7-(4-fluorobenzyl)-2,3-dihydro-1H -pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one and (2R ,5S)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine- 1-tert-butyl -carboxylate. LC-MS (M+H) + = 598. 6.

[0114] Step 5: 1-(7-(4-Fluorobenzyl)-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)-5-methyl- 2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl)ethan-1- one formate (Compound 2) Compound 2 (20 mg) as its formate was prepared in the same manner as described in Step 19 of Example 1 from (2R,5S)-4-(2-(7-(4-fluorobenzyl)-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoeth yl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1 -tert-butyl -carboxylate. 1 HNMR (400 MHz, DMSO -d6) δ 8.64 - 8.11 (s, 3H),7.85 (s, 1H), 7.3 0 - 7.19 (m, 2H), 7.18 - 7.04(m, 2H), 4.45 - 4.26 (m, 2H), 4.07 - 3.71 (m, 6H), 3.52 - 3.3 3 (m, 2H), 3.22 - 3.04 (m, 2H), 3.03 - 2.59 ( m, 7H), 2.47 - 2.37 (m, 1H), 2.21 - 2.07 (m, 1H), 2.04 - 1.91 (m, 1H), 1.89 - 1.74 (m, 1H) , 1.02 (d, J = 5.8 Hz, 3H), 0.91 - 0.74 (m, 3H ). LC-MS (M+H) + = 498.6。

[0115] Example 3: 7-(4-Fluorobenzyl)-1-(2-((2R,5R)-5-methyl- 2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl)acetyl)- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-6-carboxa midate (Compound 3)

Chemical formula

Chemical formula

[0116] Step 2: 7-(4-Fluorobenzyl)-1-(2-((2R,5R)-5-methyl -2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl)acetyl) -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-6-carbox amide formate (Compound 3) Compound 3 (30 mg) as its formate was prepared from 7-(4-fluorobenzyl)-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazine-6-carboxamide and (2R,5 S)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1- carboxylic acid tert-butyl in the same manner as described in Steps 9 and 18- 19 of Example 1. 1 HNMR (400 MHz, DMSO- d6) δ 10.26 - 8.03 (m, 4H),7.79 (s, 1H), 7.4 4 (s, 1H), 7.30 - 7.17 (m,2H), 7.14 - 6.99 ( m, 2H), 4.52 - 4.17 (m, 4H),4.13 - 3.72 (m, 4H), 3.54 - 3.29 (m, 2H), 3.17- 3.02 (m, 2H) , 3.01 - 2.76 (m, 4H), 2.73- 2.57 (m, 3H), 2 .48 - 2.35 (m, 1H), 2.21 -2.08 (m, 1H), 2.04 - 1.90 (m,1H), 1.88 - 1.71 (m, 1H), 1.05 - 0 .95 (m, 3H), 0.92 - 0.80 (m,3H). LC-MS (M+H) + = 541.6.

[0117] Example 4: 1-(6-Bromo-7-(4-fluorobenzyl)-2,3-dihydro-1H -pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)-5 -methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl)eth tan-1-one (Compound 4)

Chemical Structure

Chemical Structure

[0118] Step 2: 6-Bromo-7-(4-fluorobenzyl)-2,3-dihydro-1H-pi tert-Butyl 6-bromo-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate

Chem.

[0119] Step 3: 1-(6-Bromo-7-(4-fluorobenzyl)-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)- 5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl) ethan-1-one (Compound 4) Compound 4 as a formate salt was prepared in the same manner as described in Steps 8-9 and Steps 18-19 of Example 1 from 6-bromo-7-(4-fluorobenzyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl and tert-butyl (2R,5S)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylate. The above salt was dissolved in an aqueous solution of NaHCO3 ​​​​It was neutralized and extracted with EtOAc. The combined EtOAc layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 4 (27 mg) as the free base. 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (s, 1H), 8.25 (s, 1H), 7.28 - 7.18 (m, 2H), 7.17 - 7.09 (m, 2H), 4.52 - 4.29 (m, 2H), 4.12 - 3.89 (m, 3H ), 3.88 - 3.60 (m, 2H), 3.55 - 3.09 (m, 4H), 3.08 - 2.58 (m, 7H), 2.45 - 2.26 (m, 2H), 2.1 6 (s, 1H), 2.03 - 1.92 (m, 1H), 1.85 - 1.73 ( m, 1H), 1.01 - 0.91 (m, 3H), 0.91 - 0.74 (m, 3H). LC-MS (M+H) + = 576.5.

[0120] Example 5: 1-(7-(4-Fluorobenzyl)-6-methyl-2,3-dihydro-1H -pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)-5 -methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl)eth tan-1-one (Compound 5)

Chemical Structure

Chemical Structure

[0121] Step 2: 1-(7-(4-Fluorobenzyl)-6-methyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)- 5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl) ethan-1-one (Compound 5) Compound 5 as formate was prepared in the same manner as described in Steps 8-9 and Steps 18-19 of Example 1 from 7-(4-fluorobenzyl)-6-methyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl and (2R,5S)-2-methyl-5-(((R)-3-methylmorpholino)methyl)pi perazine-1-carboxylic acid tert-butyl. The above salt was dissolved in an aqueous solution of NaHCO3 ​ It was neutralized with [reagent] and extracted with EtOAc. The combined EtOAc layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 5 (12.6 mg) as the free base. 1 H NMR (400 MHz, DMSO-d6) δ 8.49 - 8.06 (m, 1 H), 7.21 - 7.03 (m, 4H), 4.45 - 4.24 (m, 2H), 4.10 - 3.96 (m, 2H), 3.92 - 3.84 (m, 2H), 3. 85 - 3.70 (m, 1H), 3.59 - 3.41 (m, 3H), 3.29 - 3.15 (m, 1H), 3.07 - 2.65 (m, 5H), 2.63 - 2 .53 (m, 2H), 2.31 - 2.18 (m, 4H), 2.18 - 2.06 (m, 1H), 2.01 - 1.84 (m, 2H), 1.78 - 1.65 (m , 1H), 1.00 - 0.66 (m, 6H). LC-MS (M+H) + = 51 2.3.

[0122] Example 6: 1-(6-Cyclopropyl-7-(4-fluorobenzyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5 R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1- yl)ethan-1-one (Compound 6)

Chemical Structure

[0123] Step 2: 1-(6-Cyclopropyl-7-(4-fluorobenzyl)-2,3-dih dro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R, 5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1 -yl)ethan-1-one (Compound 6) Compound 6 as a formate was prepared in the same manner as described in Steps 8-9 and Steps 18-19 of Example 1 from 6-cyclopropyl-7-(4-fluorobenzyl)-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert -butyl and (2R,5S)-2-methyl-5-(((R)-3-methylmorpholino)meth yl)piperazine-1-carboxylic acid tert-butyl and prepared from. The above salt was neutralized with an aqueous solution of NaH CO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered , and concentrated. The residue was further lyophilized to obtain compound 6 (10 mg) as the free base. 1 H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 8. 29 (s, 1H), 7.26 - 7.16 (m,2H), 7.16 - 7.06 (m, 2H), 4.43 - 4.19 (m, 2H),4.12 - 3.92 (m, 4H), 3.82- 3.72 (m, 1H), 3.53 - 3.43 (m, 3H ), 3.29 - 3.17 (m, 1H), 3.05- 2.78 (m, 2H), 2.77 - 2.63 (m, 3H), 2.61 -2.55 (m, 1H), 2.3 1 - 2.05 (m, 4H), 2.01 - 1.86(m, 2H), 1.80 - 1.66 (m, 1H), 0.95 - 0.66 (m,10H). LC-MS (M+ H) + = 538.4.

[0124] Example 7: 1-(7-(4-fluorobenzyl)-6-(trifluoromethyl)-2,3 -dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-(( (2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperaz ine-1-yl)ethan-1-one (Compound 7)

Chemical Structure

Chemical formula

[0125] Step 2: 1-(7-(4-Fluorobenzyl)-6-(trifluoromethyl)-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piper dine-1-yl)ethan-1-one (Compound 7) Compound 7 as a formate was prepared in the same manner as described in Steps 8-9 and Steps 18-19 of Example 1 from 7-(4-fluorobenzyl)-6-(trifluoromethyl)- 7-(4-Fluorobenzyl)-6-(trifluoromethyl)- 2,3-Dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-Butyl and (2R,5S)-2-methyl-5-(((R)-3-methylmorph olino)methyl)piperazine-1-carboxylic acid tert-butyl were prepared. The above salt was neutralized with an aqueous solution of N aHCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered and concentrated. The residue was further lyophilized to obtain compound 7 (26.4 mg) as the free base . 1 H NMR (400 MHz, DMSO-d6) δ 8.58 (s, 1H) , 7.27 - 7.05 (m, 4H), 4.53 - 4.35 (m, 2H), 4 .16 - 3.95 (m, 4H), 3.92 - 3.81 (m, 1H), 3.78 - 3.65 (m, 1H), 3.53 - 3.37 (m, 2H), 3.19 - 3 .07 (m, 1H), 2.97 - 2.77 (m, 4H), 2.76 - 2.57 (m, 3H), 2.47 - 2.29 (m, 2H), 2.20 - 2.09 (m , 1H), 2.01 - 1.89 (m, 1H), 1.83 - 1.73 (m, 1 H), 0.97 - 0.89 (m, 3H), 0.89 - 0.80 (m, 3H). LCMS (M+H) + = 566.3.

[0126] Example 8: 1-(7-(4-Fluorobenzyl)-6-(hydroxymethyl)-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2 R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazine -1-yl)ethan-1-one (Compound 8) [Chemical formula] Step 1: 1-(tert-Butoxycarbonyl)-7-(4-fluorobenzyl)- 6-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine 5- oxide [Chemical formula] The titled compound (1.2 g) of Step 1 was prepared from tert-butyl 7-(4-fluorobenzyl)-6-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate in the same manner as described in Step 5 of Example 1. Method for preparing 7-(4-fluorobenzyl)-6-methyl-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl. LC-MS (M+H) + = 375.2.

[0127] Step 2: 6-(Acetoxymethyl)-7-(4-fluorobenzyl)-2,3-dih dro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-but yl [Chemical formula] A solution of 1-(tert-butoxycarbonyl)-7-(4-fluorobenzyl)-6-methyl -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine 5-oxide ( 1.2 g, 3.2 mmol) in Ac2O (20 mL) was stirred at 130 °C for 3 hours. The reaction solution was cooled to room temperature, water (50 mL) was added, and the mixture was extracted with EtOAc (5m L×3). The combined organic layers were dehydrated over Na2SO4, filtered, and concentrated in vacuo to obtain a crude product (1.0 g), which was used directly in the next step without purification. Used. LC-MS (M+H) + = 417.2

[0128] Step 3: (2R,5S)-4-(2-(6-(acetoxymethyl)-7-(4-flu orobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpho olino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0129] Step 4: (2R,5S)-4-(2-(7-(4-fluorobenzyl)-6-(hydro xymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpho lino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0130] Step 5: 1-(7-(4-Fluorobenzyl)-6-(hydroxymethyl)-2,3 -dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-(( (2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperaz in-1-yl)ethan-1-one (Compound 8) Compound 8 as the formate salt was prepared in a similar manner to the method described in Step 19 of Example 1, from (2R,5S)-4-(2-(7-(4-Fluorobenzyl)-6-(hydroxymethyl) -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)- 2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)pi perazine-1-carboxylic acid tert-butyl. The above salt was neutralized with aqueous NaHCO3 and extracted with EtOAc. The combined organic layers were dried, filtered and concentrated. The residue was further lyophilized to give Compound 8 (5.7 mg) as the free base.1 HNM R (400 MHz, DMSO-d6) δ 8.34(s, 1H), 7.26 - 7 .16 (m, 2H), 7.16 - 7.06 (m,2H), 5.11 (s, 1H ), 4.49 - 4.26 (m, 4H), 4.06- 3.68 (m, 6H), 3.53 - 3.37 (m, 2H), 3.26 -2.60 (m, 10H), 2. 23 - 2.11 (m, 1H), 2.05 - 1.94(m, 1H), 1.91 - 1.80 (m, 1H), 1.10 - 1.01(m, 3H), 0.95 - 0 .76 (m, 3H). LC-MS (M+H) + = 528.3。

[0131] Example 9: 1-(7-(4-Fluorobenzyl)-6-hydroxy-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R) -5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl )ethan-1-one (Compound 9)

Chemical Structure

Chemical Structure

[0132] Step 2: 6-Acetoxy-7-(4-fluorobenzyl)-2,3-dihydro-1H -pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chemical formula

[0133] Step 3: 7-(4-Fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3 -b][1,4]oxazin-6-ol

Chemical formula

[0134] Step 4: 2-Chloro-1-(7-(4-fluorobenzyl)-6-hydroxy-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan- 1-one

Chem.

[0135] Step 5: (2R,5S)-4-(2-(7-(4-fluorobenzyl)-6-hydroxy-2,3 -dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl ))-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl )piperazine-1-carboxylic acid tert-butyl

Chem.

[0136] Step 6: 1-(7-(4-Fluorobenzyl)-6-hydroxy-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R )-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl ethyl)ethan-1-one (Compound 9) Compound 9 as formate was prepared in a similar manner to the method described in Step 19 of Example 1 from (2R,5S)-4-(2-(7-(4-fluorobenzyl)-6-hydroxy-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxo ethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine- 1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dried, filtered, and concentrated. The residue was further lyophilized to obtain Compound 9 (5.2 mg). 1 1H NMR (400 MHz, DM SO-d6) δ 9.39 (s, 1H), 8.71(s, 1H), 8.17 (s , 1H), 7.34 - 7.15 (m, 2H),7.15 - 7.02 (m, 2 H), 4.45 - 4.25 (m, 2H), 4.03- 3.65 (m, 6H), 3.57 - 3.39 (m, 3H), 3.30 - 3.05 (m, 3H), 3. 03 - 2.78 (m, 4H), 2.75 - 2.56 (m, 2H), 2.28 - 2.15 (m, 1H), 2.11 - 2.01 (m, 1H), 1.99 - 1 .83 (m, 1H), 1.20 - 1.09 (m, 3H), 0.96 - 0.79 (m, 3H). LC-MS (M + H) + = 514.5。

[0137] Example 10: 1-(7-(4-Fluorobenzyl)-3-methyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)- 5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl) ethan-1-one (Compound 10)

Chemical formula

Chemical formula

[0138] Step 2: 7-Bromo-3-methyl-1H-pyrido[2,3-b][1,4]oxadi n-2(3H)-one

Chemical formula

[0139] Step 3: 7-Bromo-3-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazine

Chemical formula

[0140] 1,4]oxazine-1-carboxylate To a solution of 7-bromo-3-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]

Chemical formula

[0141] Step 5: 7-(4-Fluorobenzyl)-3-methyl-2,3-dihydro-1H-pi rid[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chem.

[0142] Step 6: 7-(4-Fluorobenzyl)-3-methyl-2,3-dihydro-1H-pi rid[2,3-b][1,4]oxazine

Chem.

[0143] Step 7: 2-Chloro-1-(7-(4-fluorobenzyl)-3-methyl-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1- one

Chemical Structure

[0144] Step 8: (2R,5S)-4-(2-(7-(4-fluorobenzyl)-3-methyl -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)- 2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)pi perazine-1-carboxylic acid tert-butyl

Chemical Structure

[0145] Step 9: 1-(7-(4-Fluorobenzyl)-3-methyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)- 5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl) ethan-1-one (Compound 10) Compound 10 as a formate was prepared in a similar manner to the method described in Step 19 of Example 1 (2R,5S)-4-(2-(7-(4-Fluorobenzyl)-3-methyl-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoeth yl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1 -carboxylic acid tert-butyl. The above salt was neutralized with an aqueous NaHCO3 solution and extracted with EtOAc. The combined organic layers were dried, filtered, and concentrated. The residue was further lyophilized to obtain Compound 10 (8 mg) as the free base HNMR (400 M 1 Hz, DMSO-d6) δ 8.28 (s, 1H),7.85 (s, 1H), 7 .26 (s, 2H), 7.14 - 7.12 (m,2H), 4.48 (s, 1H ), 4.09 - 4.05 (m, 2H), 3.88(s, 2H), 3.55 (s ), 1H), 3.49 - 3.47 (m, 3H),3.26 - 3.20 (m, 1 , 1H), 3.49 - 3.47 (m, 3H),3.26 - 3.20 (m, 1 H), 2.93 (s, 1H), 2.79 - 2.76(m, 3H), 2.70 - 2.59 (m, 4H),2.32 - 2.26 (m, 1H), 2.16 (brs , 2H), 2.01 - 1.94 (m, 1H),1.77 - 1.72 (m, 1 H), 1.34 - 1.28 (m, 3H), 0.91- 0.82 (m, 6H). LC-MS (M+H) + = 512.0。

[0146] Example 11: 1-(6-Bromo-7-(4-fluorobenzyl)-3-methyl-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2 (R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazine -1-yl)ethan-1-one (Compound 11)

Chem.

Chem.

[0147] Step 2: 1-(6-Bromo-7-(4-fluorobenzyl)-3-methyl-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-chloro ethan-1-one

Chemical formula

[0148] Step 3: (2R,5S)-4-(2-(6-Bromo-7-(4-fluorobenzyl) -3-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino )methyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0149] Step 4: 1-(6-Bromo-7-(4-fluorobenzyl)-3-methyl-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2 (2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazine -1-yl)ethan-1-one (Compound 11) Compound 11 as a formate was prepared in a similar manner to the method described in Step 19 of Example 1, (2R,5S)-4-(2-(6-bromo-7-(4-fluorobenzyl)-3-methyl -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)- 2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)pi perazine-1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 11 (10 mg) as the free base. 1 HNM R (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 7.23 (s , 2H), 7.23 - 7.14 (m, 2H), 4.55 (s, 1H), 4.1 3 - 4.09 (m, 2H), 3.97 (s, 2H), 3.59 (brs, 1H ), 3.55 - 3.42 (m, 3H), 3.27 - 3.21 (m, 1H), 2.95 (brs, 1H), 2.77 - 2.74(m, 3H), 2.65 - 2 .59 (m, 3H), 2.30 - 2.23 (m,1H), 2.16 (brs, 2H), 2.00 - 1.95 (m, 2H), 1.76- 1.73 (m, 1H) , 1.33 (d, J = 4.0 Hz, 3H),0.86 (d, J = 8.0 H z, 6 H). LC-MS (M+H) + = 590.0, 592.0。

[0150] Example 12: 1-(6-Bromo-7-(4-fluorobenzyl)-3,3-dimethyl-2 ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2- (((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piper azin-1-yl)ethan-1-one (Compound 12)

Chemical formula

Chemical formula

[0151] Step 2: 7-Bromo-3,3-dimethyl-1H-pyrido[2,3-b][1,4]oxa zine-2-(3H)-one

Chemical Structure

[0152] Step 3: 7-Bromo-3,3-dimethyl-2,3-dihydro-1H-pyrido[2,3 -b][1,4]oxazine

Chemical Structure

[0153] Step 4: 7-Bromo-3,3-dimethyl-2,3-dihydro-1H-pyrido[2,3 -b][1,4]oxazine-1-carboxylic acid tert-butyl

Chem.

[0154] Step 5: 7-(4-Fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chem.

[0155] Step 6: 7-(4-Fluorobenzyl)-3,3-dimethyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazine

Chem.

[0156] Step 7: 6-Bromo-7-(4-fluorobenzyl)-3,3-dimethyl-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazine

Chemical formula

[0157] Step 8: 1-(6-Bromo-7-(4-fluorobenzyl)-3,3-dimethyl-2 ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2- chloroethan-1-one

Chemical formula

[0158] Step 9: (2R,5S)-4-(2-(6-Bromo-7-(4-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxa zin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmol pholino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0159] Step 10: 1-(6-Bromo-7-(4-fluorobenzyl)-3,3-dimethyl- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2 -((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)pi perazin-1-yl)ethan-1-one (Compound 12) Compound 12 as a formate was prepared in the same manner as described in Step 19 of Example 1 (2R,5S)-4-(2-(6-Bromo-7-(4-fluorobenzyl)-3,3-di (Methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl) )(2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)meth yl)piperazine-1-carboxylic acid tert-butyl was prepared. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated . The residue was further lyophilized to obtain Compound 12 (15 mg) as the free base. 1 H NMR (400 MHz, DMSO-d6) δ 8.38 (s, 1H), 7.24 (s, 2H), 7.17 - 7.13 (m, 2H), 4.14 (d, J = 15 .6 Hz, 1H), 3.97 (s, 3H), 3.57 - 3.47 (m, 4H) , 3.26 (t, J = 9.7 Hz, 1H), 2.96 (brs, 1H), 2 .78 - 2.75 (m, 4H), 2.57 - 2.55 (m, 2H), 2.30 - 2.27 (m, 1H), 2.15 (s, 2H), 2.02 - 1.88 (m, 2H), 1.77 (d, J = 12.4 Hz, 1H), 1.36 (s, 3H) , 1.30 (s, 3H), 0.91 - 0.81 (m, 6H). LC-MS (M +H) + = 604.0, 606.0.

[0160] Example 13: 2-((2R,5R)-2-(((1S,4S)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-5-methylpiperazin-1-yl) )-1-(7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-1-yl)ethan-1-one (Compound 13) [Chemistry] Step 1: (2R,5S)-5-(((1S,4S)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-4-benzyl-2-methylpiperazine- 1-carboxylic acid tert-butyl [Chemistry] The labeled compound of Step 1 (110 mg) was prepared in the same manner as the method described in Step 16 of Example 1 from (1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane salt acid salt and (2R,5R)-4-benzyl-5-(chloromethyl)-2-methylpiperazine -1-carboxylic acid tert-butyl. LC-MS (M+H) = 402 + .0

[0161] Step 2: (2R,5S)-5-(((1S,4S)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl [Chemistry] The labeled compound of Step 2 (60 mg) was prepared in the same manner as the method described in Step 17 of Example 1 from (2R,5S)-5-(((1S,4S)-2-oxa-5-azabicyclo [[2.2.1]heptan-5-yl)methyl)-4-benzyl-2-methylpiperazine-1 -carboxylic acid tert-butyl. LC-MS (M+H) = 312.0 +

[0162] Step 3: (2R,5S)-5-(((1S,4S)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-4-(2-(7-(4-fluorobenzyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0163] Step 4: 2-((2R,5R)-2-(((1S,4S)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-5-methylpiperazin-1-yl )-1-(7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-1-yl)ethan-1-one (Compound 13) Compound 13 as a formate salt was prepared in the same manner as the method described in Step 19 of Example 1 from (2R,5S)-5-(((1S,4S)-2-oxa-5-azabicyclo[2.2.1 heptan-5-yl)methyl)-4-(2-(7-(4-fluorobenzyl)-2,3 heptan-5-yl)methyl)-4-(2-(7-(4-fluorobenzyl)-2,3 ​​-Dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxo Prepared from tert-butyl 2-ethyl)-2-methylpiperazine-1-carboxylate. The above mentioned salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dried , filtered and concentrated. The residue was further lyophilized to give compound 13 (8 mg) as the free base . 1 H NMR (400 MHz, CD3OD) δ 8.44 (s, 1H) , 7.79 (s, 1H), 7.23 (s, 2H),7.04 - 7.01 (m, 2H), 4.44 - 4.41 (m, 2H), 4.31 (s, 1H), 4.10 - 4.07 (m,2H), 3.93 (brs, 3H), 3.79 (brs, 1 H), 3.56 - 3.53 (m, 1H), 3.47(d, J = 7.2 Hz, 1H), 3.37 (s, 1H), 2.99 (d,J = 12.0 Hz, 1H), 2.80 - 2.77(m, 4H), 2.64 (s, 1H), 2.55 - 2. 26 (m, 4H), 1.66 (d, J = 9.2Hz, 1H), 1.54 (d , J = 9.3 Hz, 1H), 1.03 (s,3H). LC-MS (M+H) + = 496.3.

[0164] Example 14: 2-((2R,5R)-2-(((1R,4R)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-5-methylpiperazin-1-yl )-1-(7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-1-yl)ethan-1-one (Compound 14) [Chemistry] Step 1: (2R,5S)-5-(((1R,4R)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-4-benzyl-2-methylpiperazine- 1-carboxylic acid tert-butyl [Chemistry] The labeled compound of Step 1 (100 mg) was prepared in the same manner as described in Step 16 of Example 1 from (1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane hydrochloride and (2R,5R)-4-benzyl-5-(chloromethyl)-2-methylpiperazine -1-carboxylic acid tert-butyl. LC-MS (M+H) + = 402 .0.

[0165] Step 2: (2R,5S)-5-(((1R,4R)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl [Chemistry] The labeled compound of Step 2 (60 mg) was prepared in the same manner as described in Step 17 of Example 1 from (2R,5S)-5-(((1R,4R)-2-oxa-5-azabicyclo 2.2.1]heptan-5-yl)methyl)-4-benzyl-2-methylpiperazine-1 -carboxylic acid tert-butyl. LC-MS (M+H) + = 312.0 .

[0166] Step 3: (2R,5S)-5-(((1R,4R)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-4-(2-(7-(4-fluorobenzyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl [Chemical formula] The labeled compound (30 mg) in Step 3 was prepared in the same manner as described in Step 18 of Example 1 from (2R,5S)-5-(((1R,4R)-2-oxa-5-azabicyclo [[2.2.1]heptan-5-yl)methyl)-2-methylpiperazine-1-carboxylic acid t ert-butyl and 2-chloro-1-(7-(4-fluorobenzyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one. LC-MS (M+H) = 596.0. +

[0167] Step 4: 2-((2R,5R)-2-(((1R,4R)-2-oxa-5-azabicyclo [2.2.1]heptan-5-yl)methyl)-5-methylpiperazin-1-yl )-1-(7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-1-yl)ethan-1-one (Compound 14) Compound 14 as a formate salt was prepared in the same manner as described in Step 19 of Example 1 from (2R,5S)-5-(((1R,4R)-2-oxa-5-azabicyclo[2.2.1 heptan-5-yl)methyl)-4-(2-(7-(4-fluorobenzyl)-2,3 ​​​-Dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxo Prepared from tert-butyl 2-methylpiperazine-1-carboxylate. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dried, filtered, and concentrated. The residue was further lyophilized to obtain compound 14 (5 mg) as the free base. 1 H NMR (400 MHz, CD3OD) δ 8.54 (s, 1H) , 7.80 (s, 1H), 7.22 (s, 2H), 7.04 - 7.02 (m, 2H), 4.42 (brs, 2H), 4.19 (s, 1H), 4.01 - 3. 97 (m, 2H), 3.92 (s, 2H), 3.88 - 3.87 (m, 2H) , 3.70 (d, J = 16.6 Hz, 1H), 3.46 (d, J = 7.6 Hz, 1H), 2.97 (d, J = 12.2 Hz, 1H), 2.90 - 2. 86 (m, 1H), 2.80 - 2.72 (m, 4H), 2.68 (s, 1H) , 2.54 - 2.51 (m, 2H), 2.45 (d, J = 12.3 Hz, 1 H), 2.31 (d, J = 9.4 Hz, 1H), 1.51 (s, 2H), 1 .10 - 1.05 (m, 3H). LC-MS (M+H) + = 496.3.

[0168] Example 15: (S)-4-(Difluoromethyl)-3-(((2R,5R)-1-(2- (7-(4-Fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-1-yl)-2-oxoethyl)-5-methylpiperazin-2-yl Methyl)oxazolidin-2-one (Compound 15)

Chem.

Chem.

[0169] Step 2: (2R,5R)-5-(((S)-4-(difluoromethyl)-2-oxo oxazolidin-3-yl)methyl)-2-methylpiperazine-1-carboxylic acid tert -butyl

Chem.

[0170] Step 3: (2R,5R)-5-(((S)-4-(Difluoromethyl)-2-oxo oxazolidin-3-yl)methyl)-4-(2-(7-(4-fluorobenzyl)-2 ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2- oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0171] Step 4: (S)-4-(Difluoromethyl)-3-(((2R,5R)-1-(2- (7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-1-yl)-2-oxoethyl)-5-methylpiperazin-2-yl methyl)oxazolidin-2-one (Compound 15) Compound 15 as a formate was prepared in the same manner as described in Step 19 of Example 1 from (2R,5R)-5-(((S)-4-(difluoromethyl)-2-oxooxazolidin-3-yl)methyl)-4-(2-(7-(4-fluorobenzyl)-2,3-dihydro ​​​((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpholino)methyl)-5-methylpiperazin-1-yl)-2-oxoethyl )-2-methylpiperazine-1-carboxylic acid tert-butyl was prepared. The above salt was neutralized with an aqueous solution of N aHCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered through, and concentrated. The residue was further lyophilized to obtain compound 15 (5 mg) as the free base . 1 H NMR (400 MHz, CD3OD) δ 8.54 (s, 1H), 7. 80 (s, 1H), 7.23 (s, 2H), 7.03 - 7.01 (m, 2H) , 6.09 (t, J = 54.8 Hz, 1H), 4.43 (s, 3H), 4. 29 - 4.21 (m, 2H), 4.08 (s, 1H), 3.92 (s, 2H) , 3.88 - 3.84 (m, 3H), 3.70 - 3.66 (m, 2H), 3. 21 - 3.17 (m, 2H), 2.96 - 2.82 (m, 3H), 2.59 (t, J = 11.3 Hz, 1H), 2.47 (s, 1H), 1.05 (d, J = 4.0 Hz, 3H). LC-MS (M+H) + = 534.3.

[0172] Example 16: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpholino)methyl)-5-methylpiperazin-1-yl)-1-(7-(4-fluorobenzyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )ethan-1-one (Compound 16)

Chemical Structure

Chem.

[0173] Step 2: (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino) methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chem.

[0174] Step 3: (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino) methyl)-4-(2-(7-(4-fluorobenzyl)-2,3-dihydro-1H-pyr ido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl piperazine-1-carboxylic acid tert-butyl ​ [Chemical formula] The labeled compound (20 mg) of Step 3 was prepared in the same manner as described in Step 18 of Example 1 by the method of, (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino)meth yl)-2-methylpiperazine-1-carboxylic acid tert-butyl and 2-chloro-1- (7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-1-yl)ethan-1-one. LC-MS (M+H) + = 612.3.

[0175] Step 4: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorph olino)methyl)-5-methylpiperazin-1-yl)-1-(7-(4-fluorobenzyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )ethan-1-one (Compound 16) Compound 16 as a formate salt was prepared in the same manner as described in Step 19 of Example 1 from (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino)methyl)-4 -(2-(7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine- 1-carboxylic acid tert-butyl. The above salt was neutralized with NaHCO3 and extracted with EtO Ac. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further freeze dried to obtain Compound 16 (4 mg) as the free base. 1 1H NMR (400 MHz , in CD3OD) δ 8.28 (s, 1H), 7.82 (s, 1H), 7.27 - 7.19 (m, 2H), 7.13 - 7.05 (m, 2H), 4.36 (brs , 1H), 4.30 (brs, 1H), 4.04 - 3.96 (m, 2H), 3 .85 (brs, 2H), 3.83 - 3.73 (m, 1H), 3.50 - 3. 41 (m, 1H), 3.41 - 3.34 (m, 2H), 3.04 (brs, 2 H), 2.90 - 2.82 (m, 1H), 2.73 (brs, 1H), 2.6 5 (brs, 2H), 2.59 - 2.51 (m, 2H), 2.28 - 2.18 (m, 1H), 2.18 - 2.08 (m, 1H), 2.00 - 1.88 (m , 2H), 1.21 (brs, 1H), 0.89 - 0.75 (m, 9H). L C-MS (M + H) + = 512.3。

[0176] Example 17: 1-(6-Ethoxy-7-(4-fluorobenzyl)-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R) -5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl )ethan-1-one (Compound 17)

Chemical Structure

Chemical Structure

[0177] Step 2: 5-Bromo-6-ethoxy-3-nitropyridin-2-amine

Chemical formula

[0178] Step 3: 3-Bromo-6-chloro-2-ethoxy-5-nitropyridine

Chemical formula

[0179] Step 4: 2-((5-bromo-6-ethoxy-3-nitropyridin-2-yl)oxy) ethyl acetate [ka] 3-Bromo-6-chloro-2-ethoxy-5-nitropyridine (2 g, 7.14 mmol) l) and ethyl 2-hydroxyacetate (880 mg, 8.5 mmol) at 0°C. 0% NaH (340 mg, 8.5 mmol) was added in small portions. The resulting mixture was The mixture was carefully quenched, diluted, and diluted with E The combined organic layers were dried, filtered and concentrated. The residue was evaporated onto silica gel. The product was purified by gel column chromatography (PE: EtOAc = 5:1) to give the title compound. The product was obtained (2 g, 80% yield). LC-MS (M+H) + = 349.0.

[0180] Step 5: 7-Bromo-6-ethoxy-1H-pyrido[2,3-b][1,4]oxa 2(3H)-Pyridinone [Chemical formula] The labeled compound (1 g, crude) in Step 5 was prepared in the same manner as described in Step 2 of Example 1 from 2-((5-bromo-6-ethoxy-3-nitropyridin-2-yl)oxy )ethyl acetate. The crude material was used in the next step without purification. LC- MS (M+H) + = 273.0, 275.0

[0181] Step 6: 7-Bromo-6-ethoxy-2,3-dihydro-1H-pyrido[2,3-b [1,4]oxazine [Chemical formula] 7-Bromo-6-ethoxy-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazine (500 mg, crude) was prepared in the same manner as described in Step 3 of Example 1 from 7-bromo-6-ethoxy-1H-pyrido[2,3-b][1,4]oxazin-2 (3H)-one. The crude material was used in the next step without purification. LC -MS (M+H) + = 259.0, 261.0

[0182] Step 7: 7-Bromo-6-ethoxy-2,3-dihydro-1H-pyrido[2,3-b [1,4]oxazine-1-carboxylic acid tert-butyl [Chemical formula] The labeled compound (400 mg) in Step 7 was prepared in the same manner as described in Step 4 of Example 1 Prepared from 7-bromo-6-ethoxy-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazine. LC-MS (M+H) + = 359.1, 361 .1

[0183] Step 8: tert-Butyl 6-ethoxy-7-(4-fluorobenzyl)-2,3-dihydro-1H- pyrido[2,3-b][1,4]oxazine-1-carboxylate

Chemical formula

[0184] Step 9: 6-Ethoxy-7-(4-fluorobenzyl)-2,3-dihydro-1H- pyrido[2,3-b][1,4]oxazine

Chemical formula

[0185] Step 10: 2-Chloro-1-(6-ethoxy-7-(4-fluorobenzyl)-2, (3-Dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan- 1-one

Chemical formula

[0186] Step 11: (2R,5S)-4-(2-(6-Ethoxy-7-(4-fluorobenzyl)-2,3-dihydro-1H- pyrido[2,3-b][1,4]oxazin-1-yl )-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl )piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0187] ​Step 12: 1-(6-Ethoxy-7-(4-fluorobenzyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R )-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl )ethan-1-one (Compound 17) Compound 17 as a formate salt was prepared in a similar manner to the method described in Step 19 of Example 1 (2R,5S)-4-(2-(6-Ethoxy-7-(4-fluorobenzyl)-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxo ethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine- 1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 17 (53 mg) as the free base. 1 HNMR (400 MHz, DMSO-d6) δ 8.25 (s, 1H), 7.20 - 7.17 (m , 2H), 7.10 - 7.06 (m, 2H),4.37 - 4.27 (m, 2 H), 4.18 (q, J = 8.0 Hz, 2H),3.93 - 3.70 (m, 6H), 3.50 - 3.39 (m, 3H), 3.22- 3.03 (m, 5H) , 3.00 - 2.77 (m, 4H), 2.70- 2.54 (m, 2H), 2 .05 - 2.01 (m, 1H), 1.92 -1.87 (m, 1H), 1.23 (t, J = 8.0Hz, 3H), 1.14 (d, J = 8.0 Hz, 3H) , 0.92-0.86 (m, 3H). LC-MS (M+H)+ = 542.3。

[0188] Example 18: 1-(8-(4-Fluorobenzyl)-3,4-dihydropyrido[2,3- b][1,4]oxazepin-1(2H)-yl)-2-((2R,5R)-5-methyl -2-(((R)-3-Methylmorpholino)methyl)piperazin-1-yl)ethan-1 -one (Compound 18)

Chemical Structure

Chemical Structure

[0189] Step 2: 8-Bromo-1-tosyl-1,2,3,4-tetrahydropyrido[2,3- b][1,4]oxazepine [Chemical formula] N-(3-((3,5-dibromopyridin-2-yl)oxy)propyl)-4-methyl benzenesulfonamide (2.2 g, 4.74 mmol), picolinic acid (466 mg, 3.79 mmol), CuI (1.08 g, 5.69 mmol), and K2CO3 (1. 96 g, 14.2 mmol) were added to DMSO (20 mL). The mixture was stirred at 140 °C for 3 hours under a nitrogen atmosphere. The mixture was cooled, diluted with water, and extracted with EtOAc (80 mL × 3). The combined organic layers were washed with saturated brine, dehydrated over Na2SO 4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (PE: EtOAc = 5:1) to give the title product (1.03 g, 57% yield) . LC-MS (M+H) + = 383.0, 385.0.

[0190] Step 3: 8-(4-Fluorobenzyl)-1-tosyl-1,2,3,4-tetrahydro pyrido[2,3-b][1,4]oxazepine [Chemical formula] The title compound of Step 3 (630 mg) was prepared in the same manner as described in Step 7 of Example 1 from 8-bromo-1-tosyl-1,2,3,4-tetrahydropyrido[2,3-b [1,4]oxazepine and (4-fluorobenzyl)zinc chloride (II). . LC-MS (M+H) + = 413.1.

[0191] Step 4: 8-(4-Fluorobenzyl)-1,2,3,4-tetrahydropyrido[2 ,3-b][1,4]oxazepine

Chem.

[0192] Step 5: 2-Chloro-1-(8-(4-fluorobenzyl)-3,4-dihydropyr ido[2,3-b][1,4]oxazepin-1(2H)-yl)ethan-1-one

Chem.

[0193] Step 6: (2R,5S)-4-(2-(8-(4-fluorobenzyl)-3,4-di (Hydroxypyrido[2,3 - b][1,4]oxazepin - 1(2H) - yl)-2 - oxo ethyl)-2 - methyl - 5 - (((R)-3 - methylmorpholino)methyl)piperazine - 1 - carboxylic acid tert - butyl

Chemical formula

[0194] Step 7: 1-(8-(4 - fluorobenzyl)-3,4 - dihydropyrido[2,3 - b][1,4]oxazepin - 1(2H) - yl)-2 - ((2R,5R)-5 - methyl -2 - (((R)-3 - methylmorpholino)methyl)piperazin - 1 - yl)ethan - 1 - one (Compound 18) Compound 18 as a formate was prepared in the same manner as described in Step 19 of Example 1 from (2R,5S)-4-(2-(8-(4 - fluorobenzyl)-3,4 - dihydropyrido [2,3 - b][1,4]oxazepin - 1(2H) - yl)-2 - oxoethyl)-2 - methyl - 5 - (((R)-3 - methylmorpholino)methyl)piperazine - 1 - carboxylic acid tert - butyl. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc It was extracted. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain 25 mg of Compound 18 as the free base. 1 HNMR (400 MHz, C D3OD) δ 8.14 (s, 1H), 7.77(s, 1H), 7.29 (s, 2H), 7.06 (s, 2H), 4.70 (s,1H), 4.46 (s, 1H ), 4.19 (s, 1H), 4.01 (s, 2H),3.90 (s, 1H), 3.58 (s, 1H), 3.45 (s, 1H),3.04 - 2.63 (m, 9 H), 2.52 (s, 1H), 2.34 - 2.21(m, 3H), 2.09 ( s, 1H), 1.88 - 1.77 (m, 2H),1.58 (s, 1H), 1. 01 (s, 3H), 0.84 (s, 3H). LC-MS(M+H) + = 512 .3

[0195] Example 19: 1-(7-(4-Fluorobenzyl)-2-methyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)- 5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl) ethan-1-one (Compound 19)

Chemical formula

Chemical formula

[0196] Step 2: tert-Butyl 7-bromo-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazine-1-carboxylate

Chemical Structure

[0197] Step 3: tert-Butyl 7-(4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-py rido[2,3-b][1,4]oxazine-1-carboxylate

Chemical Structure

[0198] Step 4: 7-(4-Fluorobenzyl)-2-methyl-2,3-dihydro-1H-pi rido[2,3-b][1,4]oxazine

Chemical formula

[0199] Step 5: 2-Chloro-1-(7-(4-fluorobenzyl)-2-methyl-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1- one

Chemical formula

[0200] Step 6: (2R,5S)-4-(2-(7-(4-Fluorobenzyl)-2-methyl -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)- 2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)pi perazine-1-carboxylic acid tert-butyl

Chemical formula

[0201] Step 7: 1-(7-(4-Fluorobenzyl)-2-methyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)- 5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl) ethan-1-one (Compound 19) Compound 19 as a formate (a mixture of diastereomers, 1 / 1 ratio) was used in the procedure of Example 1 In a manner similar to the method described in Step 19, (2R,5S)-4-(2-(7-(4-fluoro benzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methyl morpholino)methyl)piperazine-1-carboxylic acid tert-butyl was prepared. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dried, filtered, and concentrated. The residue was further lyophilized to obtain Compound 19 (20 mg, a mixture of diastereomers, 1 / 1 ratio) as the free base. 1 HNMR (400 MHz , DMSO-d6) δ 8.25 (s, 1H), 7.84 (d, J = 12.0 Hz, 1H), 7.30 - 7.22 (m, 2H), 7.16 - 7.09 (m, 2H), 4.77 - 4.67 (m, 1H), 4.35 - 4.19 (m, 2H ), 4.07 - 4.03 (m, 1H), 3.88 (s, 2H), 3.59 - 3.46 (m, 3H), 2.98 - 2.51 (m, 8H), 2.33 - 2.1 7 (m, 3H), 2.06 - 1.89 (m, 2H), 1.76 (t, J = 1 2.0 Hz, 1H), 1.19 - 1.12 (m, 3H), 0.91 - 0.82 (m, 6H). LC-MS (M+H) + = 512.3.

[0202] Example 19A: 1-((S)-7-(4-fluorobenzyl)-2-methyl-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R ​,5R)-5-Methyl-2-(((R)-3-methylmorpholino)methyl)piperazin- 1-yl)ethan-1-one (Compound 19A) [Chemical formula] Compound 19A as a formate salt was prepared from the chiral starting materials (S)-(1-hydroxypropan-2-yl)carbamic acid tert-butyl and (2R,5S)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl in the same manner as described in Example 19 in Step 1 . The above salt was neutralized with an aqueous NaHCO3 solution and extracted with EtOAc. The combined organic layers were dehydrated , filtered, and concentrated. The residue was further lyophilized to obtain Compound 19A (23 mg) as the free base . The above salt was neutralized with an aqueous NaHCO3 solution and extracted with EtOAc. The combined organic layers were dehydrated , filtered, and concentrated. The residue was further lyophilized to obtain Compound 19A (23 mg) as the free base . 1 1H NMR (400 MHz, DMSO-d6) δ 8.22 (s, 1 H), 7.82 (s, 1H), 7.26 - 7.22(m, 2H), 7.13 - 7.08 (m, 2H),4.74 (s, 1H), 4.32 - 4.29 (m, 1H), 4.19 - 4.16 (m, 1H), 4.05- 4.00 (m, 1H) , 3.85 (s, 2H), 3.49 - 3.45(m, 3H), 3.29 - 3 .23 (m, 1H), 2.95 (bro, 1H),2.82 - 2.64 (m, 3H), 2.59 - 2.53 (m, 2H), 2.30- 2.12 (m, 4H) , 1.95 (t, J = 12.0 Hz, 2H),1.71 (d, J = 8.0 Hz, 1H), 1.15 - 1.10 (m, 3H),0.85 - 0.83 (m, 6H). LC-MS (M+H) + = 512.5

[0203] Example 19B: 1-((R)-7-(4-Fluorobenzyl)-2-methyl-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R ,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin- 1-yl)ethan-1-one (Compound 19B) [Chemical formula] Compound 19B as a formate was prepared from (R)-(1-hydroxypropan-2-yl)carbamic acid tert-butyl and (2R,5S)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl, which are chiral starting materials in Step 1, in the same manner as described in Example 19. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 19B (15 mg) as a free base. (R)-(1-hydroxypropan-2-yl)carbamic acid tert-butyl and (2R,5S)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 19B (15 mg) as a free base. and concentrated. The residue was further lyophilized to obtain Compound 19B (15 mg) as a free base. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 19B (15 mg) as a free base. The residue was further lyophilized to obtain Compound 19B (15 mg) as a free base. 1 H NMR (400 MHz, DMSO-d6) δ 8.25 (s, 1 H), 7.83 (s, 1H), 7.27 - 7.24 (m, 2H), 7.14 - 7.09 (m, 2H), 4.67 (s, 1H), 4.38 - 4.19 (m, 3H), 3.88 (s, 2H), 3.58 - 3.51 (m, 2H), 3.31 - 3.16 (m, 2H), 3.00 (s, 1H), 2.88 - 2.72 (m, 3H), 2.63 (s, 2H), 2.32 (s, 2H), 2.17 (s, 2H ​), 2.00 (s, 2H), 1.79 - 1.76(m, 1H), 1.21 - 1.11 (m, 3H), 0.93 - 0.81 (m,6H). LC-MS (M+H ) + = 512.5。

[0204] Example 20: 1-(7'-(4-Fluorobenzyl)-1'H,3'H-spiro[cyclo propane-1,2'-pyrido[2,3-b][1,4]oxazin]-1'-yl)-2 -((2R,5R)-5-Methyl-2-(((R)-3-methylmorpholino)methyl)pi perazin-1-yl)ethan-1-one hydrochloride (Compound 20) [Chemical formula] Step 1: tert-Butyl (1-(((3,5-dibromopyridin-2-yl)oxy)methyl)cycl opropyl)carbamate [Chemical formula] The title compound (3.9 g) of Step 1 was prepared from tert-butyl (1-(hydroxymethyl)cyclopropyl)carbamate and 3,5-dibromo-2-fluoropyridine in the same manner as described in Step 1 of Example 19 by the same method. LC-MS (M+H) =423.0。 + =423.0。

[0205] Step 2: tert-Butyl 7'-bromo-1'H,3'H-spiro[cyclopropane-1,2'-pyr ido[2,3-b][1,4]oxazine]-1'-carboxylate [Chemical formula] (1-(((3,5-dibromopyridin-2-yl)oxy)methyl)cyclopropyl ) tert-Butyl carbamate (1.69 g, 4 mmol), CuI (762 mg, 4 mmol), picolinic acid (492 mg, 4 mmol), and Cs2CO3 (2.6 g, 8 mmol) were added DMSO (15 mL). The mixture was stirred at 12 0 °C for 2 h under a nitrogen atmosphere. After cooling to room temperature, the mixture was treated with water and the resulting mixture was extracted with EtOAc (80 mL × 3). The combined organic layers were washed with saturated brine , dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (PE / EtOAc = 5:1) to give the title product (360 mg, yield 27%). LC-MS (M+H) + = 341.0, 343.0.

[0206] Step 3: 7'-(4-Fluorobenzyl)-1'H,3'H-spiro[cyclopropa ne-1,2'-pyrido[2,3-b][1,4]oxazine]-1'-carboxylic acid ter t-butyl

Chemical Structure

[0207] Step 4: 7'-(4-Fluorobenzyl)-1'H,3'H-spiro[cyclopropa ne-1,2'-pyrido[2,3-b][1,4]oxazine]

Chem.

[0208] Step 5: 2-Chloro-1-(7'-(4-fluorobenzyl)-1'H,3'H-spi ro[cyclopropane-1,2'-pyrido[2,3-b][1,4]oxazine]-1' -yl)ethan-1-one

Chem.

[0209] Step 6: (2R,5S)-4-(2-(7'-(4-fluorobenzyl)-1'H, 3'H-spiro[cyclopropane-1,2'-pyrido[2,3-b][1,4]oxazi -1'-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylm orpholinomethyl)piperazine-1-carboxylic acid tert-butyl

Chem.

[0210] Step 7: 1-(7'-(4-fluorobenzyl)-1'H,3'H-spiro[cyclo propane-1,2'-pyrido[2,3-b][1,4]oxazin]-1'-yl)-2 -((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)pi perazin-1-yl)ethan-1-one hydrochloride (Compound 20) Compound 20 as formate was prepared in the same manner as the method described in Step 19 of Example 1 from (2R,5S)-4-(2-(7'-(4-fluorobenzyl)-1'H,3'H-spiro (cyclopropane-1,2'-pyrido[2,3-b][1,4]oxazin]-1'- yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)m ethyl)piperazine-1-carboxylic acid tert-butyl. The above salt was converted to NaHCO ​It was neutralized with 3 aqueous solution and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated. The residue was acidified with 1N HCl and freeze-dried to obtain Compound 20 (23 mg) as its hydrochloride. 1 H NMR (400 MHz, DMSO-d6) δ 8.35 ( s, 2H), 7.89 (d, J = 2.2 Hz,1H), 7.55 (d, J = 1.9 Hz, 1H),7.30 - 7.21 (m, 2H), 7.15 - 7.0 6 (m, 2H), 4.30 (s, 2H), 3.95- 3.76 (m, 4H), 3.47 (d, J= 11.3 Hz, 1H), 3.41 (d, J = 8.4 H z, 1H), 3.15 - 3.13 (m, 1H),2.95 (s, 1H), 2. 86 - 2.59 (m, 6H), 2.48 - 2.41(m, 1H), 2.25 (t, J = 11.2 Hz, 1H), 2.05(s, 1H), 1.87 (t, J = 10.3 Hz, 1H), 1.67 (d,J = 13.3 Hz, 1H), 1 .05 (s, 3H), 0.96 - 0.81 (m,4H), 0.66 (s, 3H ). LC-MS (M+H) + =524.4.

[0211] Example 21: 1-(7-(4-Fluorobenzyl)-6-(1-hydroxycyclopropyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)meth yl)piperazin-1-yl)ethan-1-one (Compound 21)

Chemical Structure

Chemical formula

[0212] Step 2: 1-(tert-Butoxycarbonyl)-7-(4-fluorobenzyl)- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-6-carboxylic acid

Chemical formula

[0213] Step 3: 7-(4-Fluorobenzyl)-6-(methoxy(methyl)carbamoyl) -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chemical formula

[0214] Step 4: tert-Butyl 6-acetyl-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate Under nitrogen, to a solution of tert-butyl 7-(4-fluorobenzyl)-6-(methoxy(methyl)carbamoyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate (640 mg, 1.53 mmol) in THF (20 mL) at -78 °C was added MeLi (1.4 mL, 1.6 M hexane solution, 2.24 mmol), and the mixture was stirred at the same temperature for 2 h. The reaction mixture was quenched with a saturated aqueous solution of NH4Cl, and the mixture was extracted with EtOAc (20 mL × 2). The combined organic layers were washed with saturated brine, dehydrated over Na2SO4, filtered, and concentrated by evaporation. The residue was purified by column chromatography (PE:EtOAc = 2:1) to obtain the title product (380 mg, 64% yield). H NMR (400 MHz, DMSO-d6) δ 8.14 (s, 1H ), 7.17 (dd, J = 8.1, 5.6 Hz, 2H), 7.08 (t, J = 8.9 Hz, 2H), 4.37 (s, 2H), 4.24 (s, 2H), 3. (PE:EtOAc = 2:1) to obtain the title product (380 mg, 64% yield). (PE:EtOAc = 2:1) to obtain the title product (380 mg, 64% yield). 1 H NMR (400 MHz, DMSO-d6) δ 8.14 (s, 1H ), 7.17 (dd, J = 8.1, 5.6 Hz, 2H), 7.08 (t, J = 8.9 Hz, 2H), 4.37 (s, 2H), 4.24 (s, 2H), 3. ​​​​​84 (s, 2H), 2.50 (s, 3H), 1.42(s, 9H). LC-MS (M+H) + = 387.1。

[0215] Step 5: 6-(1-((tert-butyldimethylsilyl)oxy)vinyl)-7- (4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4] oxazine-1-carboxylic acid tert-butyl

Chemical Structure

[0216] Step 6: 6-(1-((tert-butyldimethylsilyl)oxy)cyclopropyl )-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b] 1,4]oxazine

Chemical Structure

[0217] , 6H). LC-MS (M+H) + = 415.2.

[0217] Step 7: 1-(6-(1-((tert-butyldimethylsilyl)oxy)cycloprop yl)-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-1-yl)-2-chloroethan-1-one

Chem.

[0218] Step 8: (2R,5S)-4-(2-(6-(1-((tert-butyldimethylsily l)oxy)cyclopropyl)-7-(4-fluorobenzyl)-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)- 2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carbo nic acid tert-butyl

Chem.

[0219] Step 9: 1-(7-(4-Fluorobenzyl)-6-(1-hydroxycyclopropyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1-yl)ethan-1-one (Compound 21) (2R,5S)-4-(2-(6-(1-((tert-butyldimethylsilyl)oxy)cyclopropyl)-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido (2R,5S)-4-(2-(6-(1-((tert-butyldimethylsilyl)oxy)cyclopropyl)-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl -5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert -butyl (230 mg, 0.3 mmol) in methanol (3 mL) was added with HCl 1,4-dioxane solution (3 mL, 4 M) at 0 °C. The mixture was stirred at room temperature for 4 h. The solvent was distilled off, and the residue was washed with aqueous NaHCO3 solution and extracted with EA. The combined organic layers were dehydrated, filtered, and concentrated. The residue was purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain the title product (60 mg, yield 36%). (DCM:MeOH = 10:1) to obtain the title product (60 mg, yield 36%). 1 1H NMR (400 MHz, DMSO-d6) δ 8.22 (s, 1H ), 7.24 (s, 2H), 7.10 (t, J = 8.0 Hz, 2H), 6. 03 (s, 1H), 4.43 - 4.23 (m,3H), 4.15 (d, J = 15.5 Hz, 1H), 3.97 - 3.93 (m,2H), 3.74 (s, 1 H), 3.61 (d, J = 15.7 Hz, 1H),3.45 (d, J = 10 .1 Hz, 1H), 3.36 (s, 1H), 3.17 - 3.12 (m, 1H) , 2.28 - 2.78 (m, 4H), 2.66(d, J = 9.9 Hz, 2H ), 2.56 (d, J = 10.6 Hz, 1H),2.33 - 2.30 (m, 2H), 2.12 (s, 1H), 1.93 (s,1H), 1.74 (d, J = 10.7 Hz, 1H),0.90 - 0.83 (m, 10H). LC-MS (M +H) + = 554.3。

[0220] Example 22: 2 - ((2R,5R)-2 - ((((3R,5R)-3,5 - dimethylmorpho lino)methyl)-5 - methylpiperazin - 1 - yl)-1 - ((S)-7 - (4 - fluoro benzyl)-2 - methyl - 2,3 - dihydro - 1H - pyrido[2,3 - b][1,4] oxazin - 1 - yl)ethan - 1 - one (Compound 22)

Chemical Structure

[0221] Example 23: 1-((S)-2-((Benzyloxy)methyl)-7-(4-fluoroben zyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1- yl)-2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpholino Methyl)-5-methylpiperazin-1-yl)ethan-1-one (Compound 23)

Chem.

Chem.

[0222] Step 2: (S)-2-((Benzyloxy)methyl)-7-bromo-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-but yl

Chem.

[0223] Step 3: (S)-2-((Benzyloxy)methyl)-7-(4-fluorobenzyl ​​)-2,3-Dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carbo nic acid tert-butyl

Chem.

[0224] Step 4: (S)-2-((Benzyloxy)methyl)-7-(4-fluorobenzyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine

Chem.

[0225] Step 5: (S)-1-(2-((Benzyloxy)methyl)-7-(4-fluoroben zyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1- yl)-2-chloroethan-1-one

Chem.

[0226] Step 6: (2R,5S)-4-(2-((S)-2-((benzyloxy)methyl) -7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-1-yl)-2-oxoethyl)-5-(((3R,5R)-3,5 -dimethylmorpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0227] Step 7: 1-((S)-2-((benzyloxy)methyl)-7-(4-fluorobenzyl ​​​​​​-(2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpholino )methyl)-5-methylpiperazin-1-yl)ethan-1-one (Compound 23) Compound 23 as the formate salt was prepared in a similar manner to the method described in Step 19 of Example 1 from (2R,5S)-4-(2-((S)-2-((benzyloxy)methyl)-7-(4- fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxa zin-1-yl)-2-oxoethyl)-5-(((3R,5R)-3,5-dimethylmorpholino )methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 23 (210 mg , yield 47%) as the free base. HNMR (400 MHz, DMSO-d6 ) δ 8.31 (s, 1H), 7.83 (s,1H), 7.3 - 7.23 (m , 5H), 7.23 - 7.16 (m, 2H),7.15 - 7.08 (m, 2 1 H), 4.97 (brs, 1H), 4.58 -4.52 (m, 1H), 4.5 1 - 4.39 (m, 2H), 4.27 - 4.20(m, 1H), 4.19 - , 5H), 7.23 - 7.16 (m, 2H),7.15 - 7.08 (m, 2 H), 4.97 (brs, 1H), 4.58 -4.52 (m, 1H), 4.5 1 - 4.39 (m, 2H), 4.27 - 4.20(m, 1H), 4.19 - 4.11 (m, 1H), 3.88 (s, 2H),3.55 - 3.49 (m, 1 H), 3.48 - 3.41 (m, 3H), 3.31- 3.24 (m, 2H), 3.11 (brs,2H), 3.05 - 2.98 (m, 1H), 2.83 - 2.72 (m, 1H), 2.68 - 2.53 (m,4H), 2.38 (brs , 1H), 2.27 - 2.17 (m, 1H),2.08 - 1.98 (m, 1 H), 1.98 - 1.90 (m, 1H), 0.85(d, J = 4.5 Hz, 3H), 0.80 (d, J = 5.6 Hz, 6H). LC-MS(M+H) + = 632.4。

[0228] Example 24: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorph olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro benzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyr ido[2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 24)

Chemical formula

Chemical formula

[0229] Step 2: (S)-7-bromo-2,6-dimethyl-2,3-dihydro-1H-pyrido tert-Butyl [2,3-b][1,4]oxazine-1-carboxylate

Chem.

[0230] Step 3: (S)-7-(4-Fluorobenzyl)-2,6-dimethyl-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chem.

[0231] Step 4: (S)-1-(tert-Butoxycarbonyl)-7-(4-fluorobenz zyl)-2,6-dimethyl-2,3-dihydro-1H-pyrido[2,3-b][1,4] oxazine 5-oxide

Chem.

[0232] Step 5: (S)-6-(acetoxymethyl)-7-(4-fluorobenzyl)-2- methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chemical formula

[0233] Step 6: (S)-(7-(4-fluorobenzyl)-2-methyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-6-yl)methyl acetate

Chemical formula

[0234] Step 7: (S)-(1-(2-chloroacetyl)-7-(4-fluorobenzyl)- 2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6 -yl)methyl acetate

Chemical formula

[0235] Step 8: (2R,5S)-4-(2-((S)-6-(acetoxymethyl)-7-( 4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-5-(((3R,5R)-3 ,5-dimethylmorpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert -butyl

Chemical formula

[0236]

Chemical Structure

[0237] ​​​​​​​​​​​​​​Step 10: 2-((2R,5R)-2-(((3R,5R)-3,5-Dimethylmorph olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-Flu orobenzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pi rid[2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 24) Compound 24 as a formate salt was prepared in a manner similar to the method described in Step 19 of Example 1 (2R,5S)-5-(((3R,5R)-3,5-Dimethylmorpholino)methyl)-4 -(2-((S)-7-(4-Fluorobenzyl)-6-(hydroxymethyl)-2-Me thyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous NaHCO3 solution and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 24 (97m g, yield 48%) as the free base. 1 HNMR (400 MHz, DMSO-d 6) δ 8.12 (s, 1H), 7.28 - 7.17(m, 2H), 7.16 - 7.04 (m, 2H), 5.15 - 5.01(m, 1H), 4.72 (s, 1H), 4.50- 4.37 (m, 2H), 4.33 (d, J = 10.6 H z, 1H), 4.20 (d, J = 10.9 Hz,1H), 3.97 (s, 2 H), 3.94 - 3.83 (m, 1H), 3.54- 3.37 (m, 3H), 3.18 - 3.02(m, 2H), 2.97 - 2.88 (m, 1H), 2. 82 - 2.71 (m, 1H), 2.71 - 2.63(m, 2H), 2.62 - 2.53 (m, 2H), 2.31 - 2.19(m, 1H), 2.18 - 2 .05 (m, 1H), 2.02 - 1.89 (m,2H), 1.19 (d, J = 5.7 Hz, 3H), 0.92 - 0.75(m, 9H). LC-MS (M+ H) + = 556.6。

[0238] Example 25: 1-((S)-7-(4-Fluorobenzyl)-6-(hydroxymethyl) -2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 1-yl)-2-((2R,5R)-5-methyl-2-(((R)-3-methylmorpho olino)methyl)piperazin-1-yl)ethan-1-one (Compound 25)

Chemical formula

Chemical formula

[0239] Step 2: (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-6 -(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R) -3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0240] Step 3: 1-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl) -2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 1-yl)-2-((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino no)methyl)piperazin-1-yl)ethan-1-one (Compound 25)​ Compound 25 as formate was prepared in a similar manner to the method described in Step 19 of Example 1, (2R,5S)-4-(2-((S)-7-(4-Fluorobenzyl)-6-(hydroxy dimethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxo azine-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methyl morpholino)methyl)piperazine-1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dehydrated filtered, and concentrated. The residue was further lyophilized to obtain compound 25 (103 mg, yield 51 %) as the free base. 1 1H NMR (400 MHz, DMSO-d6) δ 8. 18 (s, 1H), 7.26 - 7.17 (m,2H), 7.16 - 7.06 (m, 2H), 5.12 - 5.04 (m, 1H),4.77 (s, 1H), 4 .50 - 4.37 (m, 2H), 4.37 -4.28 (m, 1H), 4.26 - 4.15 (m,1H), 4.07 - 4.00 (m, 1H), 3.97 (s , 2H), 3.56 - 3.40 (m, 3H),3.31 - 3.24 (m, 1 H), 3.05 - 2.92 (m, 1H), 2.88- 2.53 (m, 6H), 2.36 - 2.23(m, 1H), 2.23 - 2.07 (m, 2H), 2. 03 - 1.84 (m, 2H), 1.79 - 1.67(m, 1H), 1.16 (d, J = 5.9 Hz, 3H), 0.93 -0.77 (m, 6H). LC-M S (M+H) + = 542.5.

[0241] Example 26: 1-((S)-2-Cyclopropyl-7-(4-fluorobenzyl)-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-5-Methyl-2-(((R)-3-methylmorpholino)methyl)piper din-1-yl)ethan-1-one (Compound 26)

Chemical Structure

Chemical Structure

[0242] Step 2: (S)-1-Cyclopropyl-2-((3,5-dibromopyridin-2-yl l)oxy)ethan-1-amine

Chemical Structure

[0243] Step 3: (S)-(1-Cyclopropyl-2-((3,5-dibromopyridin-2- yl)oxy)ethyl)carbamic acid tert-butyl [Chemical formula] (S)-1-Cyclopropyl-2-((3,5-dibromopyridin-2- yl)oxy)ethane-1-amine (6.5 g, 19.3 mmol), (B oc)2O (6.3 g, 29 mmol), DMAP (0.1 g, 0.82 mmol), and Et3N (5.8 g, 58 mmol) in DCM (100 mL) were stirred at room temperature for 12 hours. The reaction mixture was washed with water (50 mL), extracted with DCM (50 mL × 3), the combined DCM phases were dehydrated over N a2SO4, filtered, and evaporated in vacuo. The residue was purified by silica gel chromatography -(PE:EtOAc = 5:1) to give the product (6.5 g, 77% yield ). LC-MS (M+H) + = 434.9, 436.9.

[0244] Step 4: (S)-7-Bromo-2-cyclopropyl-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl [Chemical formula] The labeled compound of Step 4 (1.2 g, yield 26%) was prepared from tert-butyl (S)-(1-cyclopropyl-2-((3,5-dibromopyridin-2-yl)oxy)ethyl)carbamate by the same method as described in Step 2 of Example 18. LC-MS (M+H) = 355.0, 357.0. +

[0245] Step 5: (S)-2-Cyclopropyl-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert- butyl [Chemical formula] The labeled compound of Step 5 (0.8 g, yield 62%) was prepared from tert-butyl (S)-7-bromo-2-cyclopropyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate and (4-fluorobenzyl)zinc chloride (II) by the same method as described in Step 7 of Example 1. LC-MS (M+H) = + 385.2.

[0246] Step 6: (S)-2-Cyclopropyl-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine [Chemical formula] ​​​​​​​​ The labeled compound of Step 6 (0.6 g, yield 100%) was prepared from tert-butyl in the same manner as described in Step 8 of Example 1. (S)-2-Cyclopropyl-7-(4-fluorobenzyl)- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid LC-MS (M+H) + = 285.0.

[0247] Step 7: (S)-2-Chloro-1-(2-cyclopropyl-7-(4-fluoroben zyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one

Chemical formula

[0248] Step 8: (2R,5S)-4-(2-((S)-2-Cyclopropyl-7-(4-f luorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0249] Step 9: 1-((S)-2-cyclopropyl-7-(4-fluorobenzyl)-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piper azin-1-yl)ethan-1-one (Compound 26) Compound 26 as a formate salt was prepared in the same manner as described in Step 19 of Example 1 from (2R,5S)-4-(2-((S)-2-cyclopropyl-7-(4-fluorobenz yl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl )piperazine-1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous NaHCO3 solution and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated The residue was further lyophilized to obtain Compound 26 (190 mg, yield 75%) as the free base. 1 1H NMR (400 MHz, DMSO-d6) δ 8.11 (s, 1H) , 7.86 (d, J = 1.8 Hz, 1H), 7.29 - 7.25 (m, 2H ), 7.15 - 7.10 (m, 2H), 4.49 - 4.46 (m, 1H), 4.24 - 4.21 (m, 1H), 4.14 - 4.00 (m, 2H), 3.8 9 (s, 2H), 3.56 - 5.48 (m, 2H), 3.40 - 3.37 ( m, 2H), 3.02 (brs, 1H), 2.85 - 2.82 (m, 1H), 2.71 (brs, 2H), 2.58 - 2.54 (m, 3H), 2.35 - 2 .09 (m, 3H), 2.04 (brs, 1H), 1.77 (s, 1H), 0 .85 (d, J = 6.1 Hz, 3H), 0.80 (brs, 4H), 0.52 (d, J = 5.9 Hz, 1H), 0.44 (s, 3H). LC-MS (M+H ) + = 538.9。

[0250] Example 27: 1 - ((S)-2 - cyclopropyl - 7 - (4 - fluorobenzyl)-2, 3 - dihydro - 1H - pyrido[2,3 - b][1,4]oxazin - 1 - yl)-2 - ( (2R,5R)-2 - ((((3R,5R)-3,5 - dimethylmorpholino)methyl)-5 - methylpiperazin - 1 - yl)ethan - 1 - one (Compound 27) [Chemical Structure] Step 1: (2R,5S)-4-(2 - ((S)-2 - cyclopropyl - 7 - (4 - f luorobenzyl)-2,3 - dihydro - 1H - pyrido[2,3 - b][1,4]oxaz in - 1 - yl)-2 - oxoethyl)-5 - ((((3R,5R)-3,5 - dimethylmorph ((S)-2-Cyclopropyl-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)(2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpholino)methyl)-5-methylpiperazin-1-yl)ethyl)methanone [Chemical formula] The title compound of Step 1 (300 mg, yield 64%) was prepared in the same manner as described in Step 18 of Example 1 from (S)-2-chloro-1-(2-cyclopropyl-7-(4- fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one and (2R,5S)-5-(((3R,5R)-3, 5-dimethylmorpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. LC-MS (M+H) = 652.9. +

[0251] Step 2: 1-((S)-2-Cyclopropyl-7-(4-fluorobenzyl)-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-2-(((3R,5R)-3,5-dimethylmorpholino)methyl)-5 -methylpiperazin-1-yl)ethan-1-one (Compound 27) Compound 27 as a formate salt was prepared in the same manner as described in Step 19 of Example 1 from (2R,5S)-4-(2-((S)-2-cyclopropyl-7-(4-fluorobenzyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-oxoethyl)-5-(((3R,5R)-3,5-dimethylmorpholino)methyl )-2-methylpiperazine-1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous NaHCO3 solution and extracted with EtOAc. The combined organic layers were dried, ​​​​Filtered and concentrated. The residue was further lyophilized to give Compound 27 (210 mg, 83% yield). It was obtained as a free base. 1 H NMR (400 MHz, DMSO-d6) δ 8.11 (s, 1H), 7.87(s, 1H), 7.28 - 7.25 (m, 2H), 7.12 (t, J = 8.8 Hz, 2H), 4.49 - 4.47 (m, 1H) , 4.23 - 4.21 (m, 1H), 4.11 - 4.09 (m, 1H), 3 .96 - 3.93 (m, 1H), 3.89 (s,2H), 3.50 - 3.48 (m, 2H), 3.25(s, 1H), 3.15 (brs, 2H), 3.04 - 3.01 (m, 1H), 2.76 - 2.68(m, 3H), 2.52 (br s, 2H), 2.40 (brs, 1H), 2.22 - 2.19 (m, 1H), 2.07 - 2.02 (m, 1H), 1.96 -1.91 (m, 1H), 0.8 4 (brs, 10H), 0.53 - 0.51 (m,2H), 0.45 (s, 2 H). LC-MS (M+H) + = 552.9.

[0252] Example 28: 1-((S)-2-Ethyl-7-(4-fluorobenzyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R, 5R)-5-Methyl-2-(((R)-3-methylmorpholino)methyl)piperazin-1 -yl)ethan-1-one (Compound 28) [Chemical Structure Diagram] Step 1: (S)-(1-((3,5-Dibromopyridin-2-yl)oxy)buta tert-Butyl (S)-(1-hydroxybutan-2-yl)carbamate

Chem.

[0253] Step 2: tert-Butyl (S)-7-bromo-2-ethyl-2,3-dihydro-1H-pyrido[2, 3-b][1,4]oxazine-1-carboxylate

Chem.

[0254] Step 3: tert-Butyl (S)-2-ethyl-7-(4-fluorobenzyl)-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate

Chem.

[0255] Step 4: (S)-2-Ethyl-7-(4-fluorobenzyl)-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazine

Chemical formula

[0256] Step 5: (S)-2-Chloro-1-(2-ethyl-7-(4-fluorobenzyl)- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan -1-one

Chemical formula

[0257] Step 6: (2R,5S)-4-(2-((S)-2-Ethyl-7-(4-fluoroben zyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1- yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)me thyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0258] Step 7: 1-((S)-2-Ethyl-7-(4-fluorobenzyl)-2,3-dih dro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R, 5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazine-1 -yl)ethan-1-one (Compound 28) Compound 28 as a formate salt was prepared in the same manner as described in Step 19 of Example 1 from (2R,5S)-4-(2-((S)-2-Ethyl-7-(4-fluorobenzyl)-2 ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2- oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piper Prepared from tert-butyl gin-1-carboxylate. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dried, filtered, and concentrated. The residue was further lyophilized to obtain compound 28 (290 mg, yield 45%) as the free base. The combined organic layers were dried, filtered, and concentrated. The residue was further lyophilized to obtain compound 28 (290 mg, yield 45%) as the free base. 1 H NMR (400 MHz, DMSO-d6) δ 8.16 (s, 1H), 7. 84 (s, 1H), 7.25 (dd, J = 8.4, 5.6 Hz, 2H), 7 .12 (t, J = 8.8 Hz, 2H), 4.45 (d, J = 11.7 Hz, 2H), 4.20 (d, J = 11.3 Hz, 1H), 4.09 (d, J = 14.4 Hz, 1H), 3.88 (s, 2H), 3.51 (s, 2H), 3. 42 (d, J = 15.9 Hz, 1H), 2.99 (s, 1H), 2.82 ( d, J = 9.9 Hz, 2H), 2.73 (d, J = 11.5 Hz, 1H), 2.71 - 2.56 (m, 4H), 2.27 (t, J = 10.8 Hz, 1H ), 2.16 (s, 2H), 2.00 (s, 1H), 1.76 (s, 1H), 1.46 (s, 2H), 0.87 - 0.81 (m, 9H). LC-MS (M + H ) + = 526.5.

[0259] Example 29: 1 - ((S)-2-benzyl-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R ,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin- 1-yl)ethan-1-one (Compound 29) [Chemical formula] Step 1: (S)-(1-((3,5-Dibromopyridin-2-yl)oxy)-3- phenylpropan-2-yl)carbamic acid tert-butyl [Chemical formula] N-[(2S)-1-Hydroxy-3-phenylpropan-2-yl]carbamic acid t ert-butyl (4.7 g, 18.9 mmol) and 3,5-dibromo-2-fluoropyr idine (4.4 g, 17.1 mmol) in THF (80 mL) were added portionwise with NaH (865 mg, 21.6 mmol, 60%) at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 5 hours under a nitrogen atmosphere. When the reaction was complete, the reaction mixture was quenched by the addition of water (60 mL). The resulting solution was extracted with ethyl acetate (60 m L×3). The organic phases were combined, washed with saturated brine, and dried over Na2SO4. The solvent was concentrated under reduced pressure, and the residue was purified by flash chromatography (DCM:EtO Ac = 3:1) to obtain the title product (9.9 g, 73% yield). LC- MS (M+H) + = 485.0.

[0260] Step 2: (S)-2-Benzyl-7-bromo-2,3-dihydro-1H-pyrido[2 ,3-b][1,4]oxazine-1-carboxylic acid tert-butyl [Chemical formula] (S)-(1-((3,5-Dibromopyridin-2-yl)oxy)-3-phenylpro tert-Butyl (1-(pyridin-2-yl)carbamate) (1.5 g, 3.1 mmol) in TH F (10 mL) solution was added with CuI (190 mg, 1.0 mmol), Cs2CO3( 2.1 g, 6.4 mmol), and DMEDA (105 mg, 1.3 mmol) at room temperature. The resulting mixture was stirred at 80 °C for 16 h under a nitrogen atmosphere. When the reaction was complete , the resulting mixture was filtered and the filter cake was washed with DCM (2 × 10 mL). The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography (PE:EtOAc = 5:1) to give the title product (260 mg, 20% yield). LC-MS (M+H ) + = 405.1.

[0261] Step 3: (S)-2-Benzyl-7-(4-fluorobenzyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chemical Structure

[0262] Step 4: (S)-2-Benzyl-7-(4-fluorobenzyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine

Chemical Structure

[0263] Step 5: (S)-1-(2-Benzyl-7-(4-fluorobenzyl)-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-chloroeth tan-1-one​​​​​ [Chem.] The labeled compound (83 mg, crude) of Step 5 was prepared in the same manner as described in Step 9 of Example 1 and (S)-2-benzyl-7-(4-fluorobenzyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine and 2-chloroacetyl chloride were used directly in the next step without purification. LC-MS (M+H) + = 411.0.

[0264] Step 6: (2R,5S)-4-(2-((S)-2-benzyl-7-(4-fluoro benzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1 -yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino) methyl)piperazine-1-carboxylic acid tert-butyl [Chem.] The labeled compound (90 mg, 64% yield) of Step 6 was prepared in the same manner as described in Step 18 of Example 1 and (S)-1-(2-benzyl-7-(4-fluorobenzyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-chloroethan-1-one and (2R,5S)-2-methyl-5-(((R)-3 -methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl were obtained. LC-MS (M+H) = 688.4. + = 688.4.

[0265] Step 7: 1-((S)-2-benzyl-7-(4-fluorobenzyl)-2,3-di (2R,5S)-4-(2-((S)-2-benzyl-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl (Compound 29) ,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin- 1-yl)ethan-1-one (Compound 29) (2R,5S)-4-(2-((S)-2-benzyl-7-(4-fluorobenzyl) -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)- 2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)pi erazine-1-carboxylic acid tert-butyl (90 mg, 0.13 mmol) in DCM ( 3 mL) was added dropwise with a solution of HCl (gas) in 1,4-dioxane (0.5 mL, 2 mmol, 4 M) at 0 °C. The resulting mixture was stirred at room temperature for 3 hours. When the reaction was complete, the resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with saturated NaHCO3 solution (10 mL). The resulting mixture was extracted with EtOAc (2 × 15 mL). The organic phases were combined, washed with saturated brine, and dried over Na2SO4. The solvent was concentrated under reduced pressure and the residue was purified by preparative HPLC under the following conditions: column XBridge Shield RP1 8 OBD column, 30 × 150 mm, 5 μm; mobile phase, acetonitrile / water (containing 10 mmol / L NH4HCO3 and 0.1% NH3·H2O), gradient of 25% - 55% in 8 minutes; detector UV 254 nm. The title compound (29 mg, yield 38%) was obtained. 8 OBD column, 30 × 150 mm, 5 μm; mobile phase, acetonitrile / water (containing 10 mmol / L NH4HCO3 and 0.1% NH3·H2O), gradient of 25% - 55% in 8 minutes; detector UV 254 nm ol / L NH4HCO3 and 0.1% NH3·H2O), gradient of 25% - 55% in 8 minutes; detector UV 254 nm The title compound (29 mg, yield 38%) was obtained. 1 1H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.92 (s, 1H), 7.34 - 7.25 (m, 2H), 7.26 (s, 3H), 7.21-7.10(m, 2H), 7.08 ( s, 2H), 4.76 (s, 1H), 4.37(d, J = 11.2 Hz, 1 H), 4.28-4.20 (m, 1H), 3.93(d, J = 2.2 Hz, 2 H), 3.69 (s, 1H), 3.51 (s,2H), 3.37 (d, J = 9.4 Hz, 2H), 3.00 (s, 1H),2.88 (s, 2H), 2.8 7-2.78 (m, 1H), 2.64-2.57 (m,4H), 2.44-2.2 8 (m, 2H), 2.29-2.07 (m, 2H),1.99 (s, 1H), 1.84 (t, J = 10.6 Hz, 1H),1.71-1.64 (m, 1H) , 0.82 (d, J = 6.2 Hz, 6H). LC-MS(M+H) + = 588 .2。

[0266] Example 30: 1-((S)-7-(4-Fluorobenzyl)-2-(p-tolyl)-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)pipera din-1-yl)ethan-1-one (Compound 30) [Chemical formula] Step 1: (S)-2-Amino-2-(p-tolyl)ethan-1-ol [Chemical formula] (S)-Amino(4-methylphenyl)acetic acid (1.9 g, 11.5 mmol) in THF (30 mL) of the solution was added dropwise with LiAlH4 (17.5 mL, 17.5 m mol, 1 M) at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at 70 °C for 16 hours under a nitrogen atmosphere . When the reaction was completed, the reaction mixture was quenched by the addition of NaOH (2 M) at 0 °C . The resulting mixture was filtered, and the filter cake was washed with THF (2 × 10 mL) . The filtrate was concentrated under reduced pressure. The aqueous layer was extracted with CH2Cl2 (2 × 30 mL). The organic phases were combined, washed with saturated brine, and dried over Na2SO4. The solvent was concentrated under reduced pressure to obtain the title compound (800 mg, yield 46%). LC-MS (M+H) + = 152.2

[0267] Step 2: (S)-(2-Hydroxy-1-(p-tolyl)ethyl)carbamic acid te rt -butyl

Chemical Structure

[0268] Step 3: (S)-(2-((3,5-dibromopyridin-2-yl)oxy)-1- (p-Tolyl)ethyl)carbamic acid tert-butyl [ka] The title compound of Step 3 (1.3 g, 86% yield) was synthesized as described in Step 1 of Example 29. In a similar manner to that described above, (S)-(2-hydroxy-1-(p-tolyl)ethyl)carba It was prepared from tert-butyl amine and 3,5-dibromo-2-fluoropyridine. C-MS (M+H) + = 485.0.

[0269] Step 4: (S)-7-Bromo-2-(p-tolyl)-2,3-dihydro-1H-pyrifos tert-Butyl do[2,3-b][1,4]oxazine-1-carboxylate [ka] The title compound of Step 4 (350 mg, 31% yield) was synthesized by hydrochloric acid addition using the procedure described in Step 2 of Example 29. (S)-(2-((3,5-dibromopyridin-2-yl)oxy)-1,2-dibromopyridin-2-yl)-1,2-dibromopyridin-2-yl (S)-(2-((3,5-dibromopyridin-2-yl)oxy ... Prepared from tert-butyl 1-(p-tolyl)ethyl)-1-(p-tolyl)-carbamate. LC -MS (M+H) + = 405.2.

[0270] Step 5: (S)-7-(4-fluorobenzyl)-2-(p-tolyl)-2,3-di Hydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert- Butyl [ka] The labeled compound of Step 5 (275 mg, yield 73%) was prepared from (S)-7-bromo-2-(p-tolyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl and (4-fluorobenzyl)zinc(II) chloride in the same manner as described in Step 3 of Example 29. LC-MS (M+H) -1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl and (4-fluorobenzyl)zinc(II) chloride. LC-MS (M+H) + = 435.3

[0271] Step 6: (S)-7-(4-Fluorobenzyl)-2-(p-tolyl)-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazine

Chemical formula

[0272] Step 7: (S)-2-Chloro-1-(7-(4-fluorobenzyl)-2-(p-t olyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-yl )ethan-1-one

Chemical formula

[0273] Step 8: (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2 -(p-tolyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxaz in-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorph olino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical Structure

[0274] Step 9: 1-((S)-7-(4-fluorobenzyl)-2-(p-tolyl)-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piper azine-1-yl)ethan-1-one (Compound 30) (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2-(p-tolyl) (aryl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-yl (R)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl D) tert-Butyl piperazine-1-carboxylate (90 mg, 0.13 mmol) A solution of CM (3 mL) was added to a 1,4-dioxane solution of HCl (gas) (0.5 mL, 2mmol, 4M) was added dropwise. The resulting mixture was stirred at room temperature for 3 hours. The reaction was complete. The mixture was concentrated under reduced pressure. The mixture was diluted with saturated NaHCO3 solution. The mixture was diluted with ethyl acetate (10 mL) and the resulting mixture was extracted with EtOAc (2×15 mL). The organic phases were combined, washed with saturated aqueous sodium chloride, and dried over Na2SO4. The solvent was removed under reduced pressure. The residue was purified by column chromatography under the following conditions: RP18 OBD column, 30×150 mm, 5 um; mobile phase, acetonitrile / water (1 0mmol / L NH4HCO3 and 0.1% NH3·H2O), 25 Purification was performed by preparative HPLC with a gradient of 0.1% to 55%; detector UV 254 nm. The compound was obtained (29 mg, yield 38%). 1 HNMR (400 MHz, DMSO- d6) δ 8.24 (s, 1H), 7.92 (s, 1H), 7.34-7.25 (m, 2H), 7.26 (s, 3H), 7.21-7.10(m, 2H), 7. 08 (s, 2H), 4.76 (s, 1H), 4.37(d, J = 11.2H z, 1H), 4.28-4.20 (m, 1H),3.93 (d, J = 2.2H z, 2H), 3.69 (s, 1H), 3.51(s, 2H), 3.37 (d, J = 9.4 Hz, 2H), 3.00 (s, 1H),2.88 (s, 2H), 2.87-2.78 (m, 1H), 2.64-2.57(m, 4H), 2.44- 2.28 (m, 2H), 2.29-2.07 (m,2H), 1.99 (s, 1H ), 1.84 (t, J = 10.6 Hz, 1H),1.71-1.64 (m, 1 H), 0.82 (d, J = 6.2 Hz, 6H). LC-MS(M+H) + = 5 88.2。

[0275] Example 31: (S)-7-(4-Fluorobenzyl)-2-methyl-1-(2-((2R ,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin- 1-yl)acetyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxa zine-6-carboxamide (Compound 31)

Chemical Structure

Chemical Structure

[0276] Step 2: (S)-1-(tert-Butoxycarbonyl)-7-(4-fluoroben zyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxa zine-6-carboxylic acid

Chemical formula

[0277] Step 3: (S)-6-Carbamoyl-7-(4-fluorobenzyl)-2-methyl- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chemical formula

[0278] Step 4: (S)-7-(4-Fluorobenzyl)-2-methyl-2,3-dihydro- 1H-pyrido[2,3-b][1,4]oxazine-6-carboxamide

Chem.

[0279] Step 5: (S)-1-(2-Chloroacetyl)-7-(4-fluorobenzyl)-2 -methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-6- carboxamide

Chem.

[0280] Step 6: (2R,5S)-4-(2-((S)-6-carbamoyl-7-(4-fluoro benzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-meth ylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0281] Step 7: (S)-7-(4-fluorobenzyl)-2-methyl-1-(2-((2R ,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazine- 1-yl)acetyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]ox azine-6-carboxamide (Compound 31) Compound 31 as a formate was prepared in a manner similar to the method described in Step 19 of Example 1. (2R,5S)-4-(2-((S)-6-carbamoyl-7-(4-fluorobenzyl )-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpho no)methyl)piperazine-1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous solution of Na HCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered and concentrated. The residue was further lyophilized to obtain compound 31 (53 mg, yield 41%) as the free base. 1 H NMR (400 MHz, DMSO-d6) δ 8.37 (s , 1H), 7.80 (s, 1H), 7.44 (s,1H), 7.30 - 7.1 7 (m, 2H), 7.08 (t, J = 8.8 Hz, 2H), 4.79 (s, 1H), 4.39 -4.26 (m, 4H), 4.07(d, J = 15.6 Hz , 1H), 3.55 - 3.45 (m, 3H),3.30 - 3.17 (m, 1 H), 2.97 (t, J = 9.7 Hz, 1H),2.83 - 2.72 (m, 4H), 2.60 - 2.56 (m, 2H), 2.37- 2.06 (m, 4H) , 1.96 (t, J = 9.4 Hz, 1H),1.74 (d, J = 11.6 Hz, 1H), 1.19 (d, J = 6.4 Hz,3H), 0.86 (d, J = 5.8 Hz, 6H). LC-MS (M+H) + = 555.5.

[0282] Example 32: (S)-1-(2-((2R,5R)-2-(((3R,5R)-3,5- Dimethylmorpholino)methyl)-5-methylpiperazin-1-yl)acetyl)-7-( 4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazine-6-carboxamide (Compound 32)

Chemical Structure

Chemical Structure

[0283] Step 2: (S)-1-(2-((2R,5R)-2-(((3R,5R)-3,5- Dimethylmorpholino)methyl)-5-methylpiperazin-1-yl)acetyl)-7-( ​(4-Fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazine-6-carboxamide (Compound 32) Compound 32 as a formate salt was prepared in a similar manner to the method described in Step 19 of Example 1 (2R,5S)-4-(2-((S)-6-carbamoyl-7-(4-fluorobenzyl )-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-yl)-2-oxoethyl)-5-(((3R,5R)-3,5-dimethylmorpho linyl)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dehydrated, filtered, and concentrated. The residue was further lyophilized to obtain Compound 32 (15 mg, yield 17%) as the free base. 1 1H NMR (400 MHz, DMSO-d6) δ 8.32 (s, 1H), 7.80 (s, 1H), 7.44 (s, 1H), 7. 22 (s, 2H), 7.08 (t, J = 8.8 Hz, 2H), 4.75 (s , 1H), 4.43 - 4.21 (m, 4H), 3.96 (d, J = 15.4 Hz, 1H), 3.52 (d, J = 15.3 Hz, 1H), 3.42 (d, J = 9.2 Hz, 2H), 3.08 (s, 2H), 2.94 (d, J = 10 .5 Hz, 1H), 2.77 (d, J = 8.4 Hz, 1H), 2.71 - 2 .55 (m, 5H), 2.30 - 2.18 (m, 2H), 1.96 (d, J = 8.6 Hz, 1H), 1.24 - 1.18 (m, 4H), 0.87 (d, J = 5.8 Hz, 3H), 0.83 (d, J= 6.0 Hz, 6H). LC- MS (M+H) + = 569.5.

[0284] Example 33: 1-((S)-7-(4-fluorobenzyl)-2-isobutyl-2,3- Dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2 R,5R)-5-Methyl-2-(((R)-3-methylmorpholino)methyl)piperazine -1-yl)ethan-1-one (compound 33) [ka] Step 1: (S)-(1-((3,5-dibromopyridin-2-yl)oxy)-4- tert-Butyl methylpentan-2-ylcarbamate [ka] The title compound of Step 1 (6 g, 83% yield) was synthesized by the method described in Step 1 of Example 29. The same procedure was used to obtain (S)-(1-hydroxy-4-methylpentan-2-yl)carbamate. It was prepared from tert-butyl phosphate and 3,5-dibromo-2-fluoropyridine. -MS (M+H) + = 451.0.

[0285] Step 2: (S)-7-Bromo-2-isobutyl-2,3-dihydro-1H-pyrido[ 2,3-b][1,4]oxazine-1-carboxylate tert-butyl [ka] The title compound of Step 2 (760 mg, 15% yield) was synthesized by hydrochloric acid addition using the procedure described in Step 2 of Example 29. Prepared in a similar manner to the method described above from tert-butyl (S)-(1-((3,5-dibromopyridin-2-yl)oxy)-4-methylpentan-2-yl)carbamate. LC-MS (M+H) + = 371.1.

[0286] Step 3: (S)-7-(4-Fluorobenzyl)-2-isobutyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chemical formula

[0287] Step 4: (S)-7-(4-Fluorobenzyl)-2-isobutyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine

Chemical formula

[0288] Step 5: (S)-2-Chloro-1-(7-(4-fluorobenzyl)-2-isobutyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one ethan-1-one

Chemical Structure

[0289] Step 6: (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2-isobutyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical Structure

[0290] Step 7: 1-((S)-7-(4-Fluorobenzyl)-2-isobutyl-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2 (R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazine -1-yl)ethan-1-one (Compound 33) Compound 33 (13 mg, yield 6%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2-iso butyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl from (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2-isobutyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl 1 HNMR (300 MHz, DMSO-d6) δ 8.09 (s, 1H), 7.83 (d, J = 2 .1 Hz, 1H), 7.28-.16 (m, 2H),7.16-7.04 (m, 2H), 4.65-4.11 (m, 3H), 3.99(d, J = 15.9 Hz , 1H), 3.86 (s, 2H), 2.92 (s,2H), 2.82-2.65 (m, 4H), 2.58-2.49(m, 2H), 2.36-2.14 (m, 3 H), 2.11-1.87 (m, 3H), 1.66(s, 3H), 1.28 (d , J = 34.5 Hz, 3H), 0.91-0.80(m, 12H). LC-MS (M+H) + = 554.4

[0291] Example 34: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpho olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro benzyl)-2-(methoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b [1,4]oxazin-1-yl)ethan-1-one (Compound 34)

Chemical formula

Chemical formula

[0292] Step 2: (S)-2-((benzyloxy)methyl)-7-chloro-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-but yl

Chemical formula

[0293] Step 3: (S)-2-((Benzyloxy)methyl)-7-(4-fluorobenzyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carbo nic acid tert-butyl

Chemical Structure

[0294] Step 4: (S)-7-(4-Fluorobenzyl)-2-(hydroxymethyl)-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid te rt-butyl

Chemical Structure

[0295] Step 5: (R)-7-(4-Fluorobenzyl)-2-(((methylsulfonyl)oxy)methyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-carboxylic acid tert-butyl

Chemical Structure

[0296] Step 6: (R)-(7-(4-Fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-2-yl)methyl methanesulfonate

Chemical Structure

[0297] Step 7: (S)-7-(4-Fluorobenzyl)-2-(methoxymethyl)-2,3 -dihydro-1H-pyrido[2,3-b][1,4]oxazine

Chemical formula

[0298] Step 8: (S)-2-Chloro-1-(7-(4-fluorobenzyl)-2-(methoxy​ (dimethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1 -yl)ethan-1-one

Chemical Structure

[0299] Step 9: (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino) methyl)-4-(2-((S)-7-(4-fluorobenzyl)-2-(methoxymethyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl) -2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical Structure

[0300] Step 10: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorph olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-flu orobenzyl)-2-(methoxymethyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-1-yl)ethan-1-one (Compound 34) Compound 34 (30 mg, yield 85%) was prepared in the same way as described in Step 7 of Example 29 from (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino)me thyl)-4-(2-((S)-7-(4-fluorobenzyl)-2-(methoxymethyl) -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)- 2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. Obtained. 1 H NMR (300 MHz, DMSO-d6) δ 8.27 (s, 1H), 7.84 (s, 1H), 7.30 - 7.18 (m, 2H), 7.18 - 7.05 (m, 2H), 4.88 (s, 1H), 4.50 (d, J = 11.4 Hz, 1H), 4.20 (d, J = 11.4, 2.7 Hz, 1H), 4.08 (d, J = 15.0 Hz, 1H), 3.87 (s, 2H), 3.53 - 3.43 (m , 2H), 3.41 - 3.32 (m, 3H), 3.23 (s, 3H), 3.21 - 3.09 (m, 2H), 3.06 - 2.96 (m, 1H), 2.87 - 2.62 (m, 4H), 2.59 - 2.50 (m, 2H), 2.46 - 2.39 (m, 1 H), 2.30 - 2.17 (m, 1H), 2.17 - 2.04 (m, 1H), 2 .01 - 1.88 (m, 1H), 0.91 - 0.79 (m, 9H). LC-MS ( M + H) + = 556.3.

[0301] Example 35: 1 - ((S)-7-(4 - fluorobenzyl)-2-(methoxymethyl)- 2,3 - dihydro - 1H - pyrido[2,3 - b][1,4]oxazin - 1 - yl)-2 -((2R,5R)-5 - methyl - 2 - (((R)-3 - methylmorpholino)methyl)pi perazin - 1 - yl)ethan - 1 - one (Compound 35)

Chemical Structure

Chemical Structure

[0302] Step 2: 1-((S)-7-(4-Fluorobenzyl)-2-(methoxymethyl)- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2 -((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)pi perazin-1-yl)ethan-1-one (Compound 35) Compound 35 (57 mg, yield 59%) was prepared in the same way as described in Step 7 of Example 29 from (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2- (methoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxa zin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorph olino)methyl)piperazine-1-carboxylic acid tert-butyl. 1 HN MR (300 MHz, chloroform-d) δ 8.48(s, 1H), 7.84 ( s, 1H), 7.23-7.10 (m, 2H),7.06-6.92 (m, 2H ), 5.11 (t, J = 7.5 Hz, 1H),4.66-4.53 (m, 2H ), 4.23 (dd, J = 11.3, 2.8Hz, 1H), 3.89 (s, 2H), 3.76-3.51 (m, 3H), 3.51-3.29(m, 2H), 3.30 (s, 3H), 3.27-3.09 (m,2H), 3.09-2.88 (m, 2H), 2.87-2.74 (m, 2H),2.78-2.65 (m, 1 H), 2.62-2.54 (m, 1H), 2.53(s, 2H), 2.39-2 .28 (m, 1H), 2.23-2.02 (m,2H), 1.96-1.85 ( m, 1H), 1.02 (d, J = 6.3 Hz, 3H), 0.96 (d, J = 6.3 Hz, 3H). LC-MS(M+H) + = 542.3。

[0303] Example 36: 2-((2R,5R)-2-(((3R,5R)-3,5-Dimethylmorpho olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-2-(ethoxymeth yl)-7-(4-fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b [1,4]oxazin-1-yl)ethan-1-one (Compound 36)

Chemical Structure

Chemical Structure

[0304] Step 2: (S)-2-Chloro-1-(2-(ethoxymethyl)-7-(4-fluoro benzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1 -yl)ethan-1-one

Chemical Structure

[0305] Step 3: (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino) methyl)-4-(2-((S)-2-(ethoxymethyl)-7-(4-fluorobenzyl )-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl) -2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0306] Step 4: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpho lino)methyl)-5-methylpiperazin-1-yl)-1-((S)-2-(ethoxymeth (R)-7-(4-Fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b [1,4]oxazin-1-yl)ethan-1-one (Compound 36) Compound 36 (21 mg, yield 15%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-5-(((3R,5R)-3,5-Dimethylmorpholino)methyl )-4-(2-((S)-2-(Ethoxymethyl)-7-(4-fluorobenzyl)- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2 -oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. 1 H NMR (300 MHz, chloroform-d) δ 8.50 (s, 1H), 7.83 (s, 1H), 7.21 - 7.11 (m, 2H), 6.98 (t, J = 8.5 Hz, 2H), 5.09 (t, J = 7.4 Hz, 1H), 4.62 (d, J = 11.3 Hz, 1H), 4.52 (d, J = 13.7 Hz, 1H ), 4.24 (d, J = 11.4, 2.8 Hz, 1H), 3.89 (s, 2 H), 3.63 (d, J = 10.9, 3.0 Hz, 2H), 3.56 - 3.4 6 (m, 1H), 3.51 - 3.37 (m, 3H), 3.42 - 3.26 (m, 2H), 3.21 (d, J = 10.5 Hz, 1H), 3.12 (d, J = 1 3.7 Hz, 1H), 2.98 - 2.87 (m, 1H), 2.86 - 2.77 ( m, 3H), 2.72 (d, J = 11.5, 2.9 Hz, 1H), 2.58 - 2.41 (m, 2H), 2.18 - 2.00 (m, 2H), 1.14 (t, J ​= 7.0 Hz, 3H), 1.02 (d, J = 6.2 Hz, 3H), 0.96 (d, J = 6.4 Hz, 6H). LC-MS (M+H) + = 570.5

[0307] Example 37: 1-((S)-2-(ethoxymethyl)-7-(4-fluorobenzyl)- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2 -((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)pi perazin-1-yl)ethan-1-one (Compound 37)

Chemical Structure

Chemical Structure

[0308] Step 2: 1-((S)-2-(Ethoxymethyl)-7-(4-fluorobenzyl)- 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2 -((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)pi perazin-1-yl)ethan-1-one (Compound 37) Compound 37 (40 mg, yield 47%) was prepared in the same way as described in Step 7 of Example 29 from (2R,5S)-4-(2-((S)-2-(ethoxymethyl)-7-(4- fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxa zin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorph olino)methyl)piperazine-1-carboxylic acid tert-butyl. 1 HN MR (300 MHz, DMSO-d6) δ 8.27(s, 1H), 7.83 ( s, 1H), 7.29-7.17 (m, 2H),7.17-7.02 (m, 2H ), 4.79 (s, 1H), 4.48 (dd,J = 11.5, 1.3 Hz, 1H), 4.27-4.10 (m, 2H), 3.86(s, 2H), 3.60- 3.47 (m, 2H), 3.44-3.17 (m,7H), 3.07-2.92 (m, 2H), 2.86-2.56 (m, 5H),2.44-2.25 (m, 2 H), 2.22-2.16 (m, 1H), 2.15-2.03(m, 1H), 1 .87-1.77 (m, 1H), 1.02 (t,J = 7.0 Hz, 3H), 0 .93 (d, J = 6.2 Hz, 3H), 0.81(s, 3H). LC-MS ( M+H)+ = 556.3。

[0309] Example 38: 1-((S)-7-(4-Fluorobenzyl)-2-(isopropoxymethyl yl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl yl)piperazin-1-yl)ethan-1-one (Compound 38)

Chemical formula

Chemical formula

[0310] Step 2: (S)-2-chloro-1-(7-(4-fluorobenzyl)-2-(isop ropoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxaz in-1-yl)ethan-1-one

Chemical formula

[0311] Step 3: (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2 -(isopropoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-me thylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0312] Step 4: 1-((S)-7-(4-fluorobenzyl)-2-(isopropoxymeth yl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)meth (Piperazin-1-yl)ethan-1-one (Compound 38) Compound 38 (63 mg, yield 73%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2- (isopropoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4] oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methyl morpholino)methyl)piperazine-1-carboxylic acid tert-butyl. 1 H NMR (300 MHz, chloroform-d) δ8.50 (s, 1H), 7.8 3 (s, 1H), 7.20 - 7.09 (m, 2H),7.04 - 6.91 (m, 2H), 4.99 - 4.93 (m, 1H), 4.66 - 4.52(m, 2H), 4.23 (d, J = 11.3, 2.9 Hz,1H), 3.88 (s, 2H), 3.74 - 3.34(m, 6H), 3.26 - 3.01 (m, 2H), 3.00 - 2.88 (m, 1H), 2.87 - 2.67 (m,3H), 2.62 - 2.50 (m, 2H), 2.33(s, 1H), 2.25 - 2.03 (m, 2H), 2 .01 (s, 1H), 1.95 - 1.85 (m,1H), 1.09 (d, J = 6.1 Hz, 3H), 1.06 - 0.99 (m,6H), 0.96 (d, J = 6.3 Hz, 3H). LC-MS (M+H) + = 570.3.

[0313] Example 39: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpho lino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino)methyl)-4-(2-((S)-7-(4-fluorobenzyl)-2-(isopropoxymethyl)-2,3-dihydro-1H-pyrido[2, 3-b][1,4]oxazin-1-yl)ethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl (Compound 39)

Chemical formula

Chemical formula

[0314] Step 2: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorph olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro robenzyl)-2-(isopropoxymethyl)-2,3-dihydro-1H-pyrido[2, ​(3-b)[1,4]oxazin-1-yl)ethan-1-one (Compound 39) Compound 39 (60 mg, yield 72%) was prepared in the same manner as described in Step 7 of Example 29 by the method of (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino)me thyl)-4-(2-((S)-7-(4-fluorobenzyl)-2-(isopropoxym thyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester from. 1 1H NMR (300 MHz, chloroform-d) δ 8.51 (s, 1H), 7.83 (s, 1H), 7.20 - 7.09 (m, 2H), 7.03 - 6.91 (m, 2H), 5.05 - 4.96 (m,1H), 4.65 - 4.47 (m, 2H), 4.27 - 4.19 (m, 1H),3.88 (s, 2H), 3. 68 - 3.57 (m, 2H), 3.55 - 3.36 (m, 3H), 3.36 - 3. 26 (m, 2H), 3.22 (d, J = 9.8 Hz, 1H), 3.10 (d , J = 13.7 Hz, 1H), 2.98 - 2.87 (m, 1H), 2.85 - 2.66 (m, 3H), 2.57 - 2.44 (m,2H), 2.16 - 2.02 (m, 2H), 1.10 (d, J = 6.1 Hz,3H), 1.05 - 0.99 (m, 6H), 0.96 (d, J = 6.4 Hz,6H). LC-MS (M+H ) + = 584.3.

[0315] Example 40: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpho (R)-methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro benzyl)-2-(phenoxymethyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-1-yl)ethan-1-one (Compound 40)

Chemical Structure

Chemical Structure

[0316] Step 2: (S)-2-chloro-1-(7-(4-fluorobenzyl)-2-(pheno xy methyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 1-yl)ethan-1-one

Chemical Structure

[0317] Step 3: (2R,5S)-5-(((3R,5R)-3,5-Dimethylmorpholino) methyl)-4-(2-((S)-7-(4-Fluorobenzyl)-2-(phenoxymeth yl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical Structure

[0318] Step 4: 2-((2R,5R)-2-(((3R,5R)-3,5-Dimethylmorph olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-Fluoro benzyl)-2-(phenoxymethyl)-2,3-dihydro-1H-pyrido[2,3- b][1,4]oxazin-1-yl)ethan-1-one (Compound 40) Compound 40 (48 mg, 56% yield) was prepared in the same manner as described in Step 7 of Example 29 The method of (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino)meth yl)-4-(2-((S)-7-(4-fluorobenzyl)-2-(phenoxymethyl ))-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl) -2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl was prepared . 1 H NMR (300 MHz, chloroform-d) δ 8.47 (s, 1H ), 7.84 (s, 1H), 7.30 - 7.19 (m, 2H), 7.19 - 7. 09 (m, 2H), 7.03 - 6.91 (m, 3H), 6.85 - 6.76 (m , 2H), 5.41 (s, 1H), 4.78 - 4.61 (m, 2H), 4.38 - 4.30 (m, 1H), 4.13 - 3.95 (m, 2H), 3.88 (s, 2 H), 3.65 - 3.55 (m, 2H), 3.34 - 3.23 (m, 2H), 3 .23 - 3.10 (m, 2H), 2.97 - 2.85 (m, 1H), 2.76 ( s, 4H), 2.57 - 2.43 (m, 2H), 2.17 - 2.01 (m, 2H ), 1.01 (d, J = 6.2 Hz, 3H), 0.89 (d, J = 6.4 Hz, 6H). LC-MS (M+H) + = 618.5.

[0319] Example 41: 1 - ((S)-7-(4-fluorobenzyl)-2-(phenoxymethyl) - 2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)- 2 - ((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl) piperazin-1-yl)ethan-1-one (Compound 41) [Chemical formula] Step 1: (2R,5S)-4-(2-((S)-7-(4-Fluorobenzyl)-2 -(Phenoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]ox azine-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methyl morpholino)methyl)piperazine-1-carboxylic acid tert-butyl [Chemical formula] The labeled compound of Step 1 (97 mg, yield 46%) was prepared in the same manner as described in Step 18 of Example 1 from (S)-2-chloro-1-(7-(4-fluorobenzyl)-2 -(Phenoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]ox azine-1-yl)ethan-1-one and (2R,5S)-2-methyl-5-(((R )-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl. LC-MS (M+H) = 704.5. +

[0320] Step 2: 1-((S)-7-(4-Fluorobenzyl)-2-(phenoxymethyl) -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)- 2-((2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl) piperazin-1-yl)ethan-1-one (Compound 41) Compound 41 (45 mg, yield 49%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2- (Phenoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl was prepared from (Sazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl. It was prepared from 1 1 H NMR (300 MHz, chloroform-d) δ 8.47 (s, 1H), 7.84 (s, 1H), 7.30 - 7.19 (m, 2H), 7.19 - 7.09 (m, 2 H), 7.03 - 6.91 (m, 3H), 6.87 - 6.75 (m, 2H), 5 .41 (t, J = 7.5 Hz, 1H), 4.74 (dd, J = 11.5, 1 .5 Hz, 2H), 4.38 - 4.30 (m, 1H), 4.11 - 3.92 (m , 2H), 3.88 (s, 2H), 3.73 - 3.63 (m, 1H), 3.63 - 3.48 (m, 2H), 3.21 - 3.09 (m, 2H), 3.06 - 2.86 (m, 2H), 2.82 - 2.68 (m, 3H), 2.56 (t, J = 10. 9 Hz, 1H), 2.48 (s, 1H), 2.29 (s, 1H), 2.20 - 1.98 (m, 3H), 1.96 - 1.84 (m, 1H), 1.01 (d, J = 6.3 Hz, 3H), 0.86 (d, J = 6.3 Hz, 3H). LC-MS (M + H) + = 604.4.

[0321] Example 42: 1 - ((S)-2-Cyclohexyl-7-(4-fluorobenzyl)-2, 3-Dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-5-Methyl-2-(((R)-3-methylmorpholino)methyl)piperidine (2-Methyl-1H-indol-1-yl)ethan-1-one (Compound 42) [Chemistry] Step 1: 3-Bromo-2-fluoro-5-(4-fluorobenzyl)pyridine [Chemistry] To a solution of 3,5-dibromo-2-fluoropyridine (9.5 g, 37.2 mmol), XantPhos (650 mg, 1.1 mmol), and Pd2(dba)3 (70 mg, 0.07 mmol) in THF (75 mL) was added dropwise a THF solution (74.6 mL, 37.3 mmol, 0.5 M) of chloro[(4-fluorophenyl)methyl]zinc at room temperature over 5 minutes. The resulting mixture was stirred at 40 °C for 6 hours under a nitrogen atmosphere. When the reaction was complete, the resulting mixture was concentrated under reduced pressure, and the residue was purified by flash column chromatography (PE:EtOAc = 20:1) to obtain the title compound (2.8 g, yield 31%). LC-MS (M+H) = 284.0. (PE:EtOAc = 20:1) to obtain the title compound (2.8 g, yield 31%). LC-MS (M+H) + = 284.0.

[0322] Step 2: (S)-(2-((3-Bromo-5-(4-fluorobenzyl)pyridin-2-yl)oxy)-1-cyclohexylethyl)carbamic acid tert-butyl [Chemistry] The title compound of Step 2 (270 mg, yield 63%) was prepared in the same manner as described in Step 1 of Example 29 using 3-bromo-2-fluoro-5-(4-fluorobenzyl)pyridine and (S)-(1-cyclohexyl-2-hydroxyethyl)carbamic acid ter t-butyl and (S)-(1-cyclohexyl-2-hydroxyethyl)carbamic acid tert-butyl.​​​​​​ Prepared from t-butyl. LC-MS (M+H) + = 507.1

[0323] Step 3: (S)-2-Cyclohexyl-7-(4-fluorobenzyl)-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert- butyl

Chemical formula

[0324] Step 4: (S)-2-Cyclohexyl-7-(4-fluorobenzyl)-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazine

Chemical formula

[0325] Step 5: (S)-2-Chloro-1-(2-cyclohexyl-7-(4-fluorobenz Dihydro-2,3-pyrido[2,3-b][1,4]oxazine-1-yl ETHAN-1-ONE [ka] The title compound of Step 5 (105 mg, 88% yield) was synthesized as described in Step 9 of Example 1. In a manner similar to that of (S)-2-cyclohexyl-7-(4-fluorobenzyl)- 2,3-Dihydro-1H-pyrido[2,3-b][1,4]oxazine and 2-chloroa Prepared from cetyl chloride. LC-MS (M+H) + = 403.2.

[0326] Step 6: (2R,5S)-4-(2-((S)-2-cyclohexyl-7-(4-fluorophenyl) (fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazide (R)-3-methylmorpholine tert-Butyl methylpiperazine-1-carboxylate [ka] The title compound of Step 6 (40 mg, 30% yield) was synthesized as described in Step 18 of Example 1. In a similar manner to that described above, (S)-2-chloro-1-(2-cyclohexyl-7-(4-fluorophenyl) (fluorobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazide (2R,5S)-2-methyl-5-(((R)-3 Prepared from tert-butyl (methylmorpholino)methyl)piperazine-1-carboxylate LC-MS (M+H) + =680.5.

[0327] Step 7: 1-((S)-2-Cyclohexyl-7-(4-fluorobenzyl)-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-5-Methyl-2-(((R)-3-methylmorpholino)methyl)pipera zin-1-yl)ethan-1-one (Compound 42) Compound 42 (12 mg, yield 24%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-4-(2-((S)-2-Cyclohexyl-7-(4-flu orobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholi no)methyl)piperazine-1-carboxylic acid tert-butyl. 1 HNMR (300 MHz, chloroform-d) δ 8.26 (s,1H), 7.83 (s, 1H), 7.18 - 7.08 (m, 2H), 7.04 - 6.92(m, 2H), 4.67 (d, J = 11.3 Hz, 1H),4.53 - 4.30 (m, 2H) , 4.17 (dd, J = 11.3, 2.7 Hz,1H), 3.90 (s, 2 H), 3.74 - 3.45 (m, 3H), 3.19(s, 1H), 3.04 (d , J = 8.4 Hz, 2H), 2.89 - 2.65(m, 4H), 2.53 (d , J = 7.7 Hz, 2H), 2.34 (s,3H), 2.15 (s, 2H) , 2.02 - 1.89 (m, 3H), 1.79 - 1.70(m, 1H), 1.3 9 (s, 1H), 1.26 - 0.63 (m, 12H). LC-MS(M+H) + = 580.5.

[0328] Example 43: 1-((S)-2-Cyclohexyl-7-(4-fluorobenzyl)-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-2-(((3R,5R)-3,5-Dimethylmorpholino)methyl)-5 -methylpiperazin-1-yl)ethan-1-one (Compound 43)

Chemical formula

Chemical formula

[0329] Step 2: 1-((S)-2-Cyclohexyl-7-(4-fluorobenzyl)-2, 3-Dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-( (2R,5R)-2-(((3R,5R)-3,5-Dimethylmorpholino)methyl)-5 -methylpiperazin-1-yl)ethan-1-one (Compound 43) Compound 43 (15 mg, yield 27%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-4-(2-((S)-2-Cyclohexyl-7-(4-flu orobenzyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-yl)-2-oxoethyl)-5-(((3R,5R)-3,5-Dimethylmorph olino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl . 1 H NMR (300 MHz, chloroform-d) δ 8.26 (s, 1H), 7 .84 (s, 1H), 7.14 (t, J = 7.1Hz, 2H), 6.98 ( t, J = 8.6 Hz, 2H), 4.68 (d,J = 11.3 Hz, 1H), 4.38 (s, 2H),4.22 - 4.12 (m, 1H), 3.90 (s, 2 H), 3.62 (d, J = 10.9 Hz, 2H),3.36 - 3.21 (m, 3H), 3.13 - 2.96 (m, 1H), 2.80(s, 4H), 2.69 ( d, J = 11.3 Hz, 1H), 2.47 (d,J = 10.1 Hz, 2H) , 2.25 - 1.92 (m, 4H), 1.77 - 1.53(m, 3H), 1.4 9 - 1.36 (m, 1H), 1.15 - 1.11 (m,2H), 1.06 - 0.8 6 (m, 13H). LC-MS (M+H) + = 594.3

[0330] Example 44: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpho olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro benzyl)-2-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazin-1-yl)ethan-1-one (Compound 44)

Chemical Structure

Chemical Structure

[0331] Step 2: tert-Butyl (S)-7-chloro-2-phenyl-2,3-dihydro-1H-pyrido[2 ,3-b][1,4]oxazine-1-carboxylate

Chemical Structure

[0332] Step 3: (S)-7-(4-Fluorobenzyl)-2-phenyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid tert-butyl

Chem.

[0333] Step 4: (S)-7-(4-Fluorobenzyl)-2-phenyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine

Chem.

[0334] Step 5: (S)-2-Chloro-1-(7-(4-fluorobenzyl)-2-phenyl -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)eth tan-1-one [Chemical formula] The labeled compound of Step 5 (350 mg, yield 51%) was prepared in the same manner as described in Step 9 of Example 1 from (S)-7-(4-fluorobenzyl)-2-phenyl-2,3 -dihydro-1H-pyrido[2,3-b][1,4]oxazine and 2-chloroacetyl chloride. LC-MS (M+H) + = 397.1.

[0335] Step 6: (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino) methyl)-4-(2-((S)-7-(4-fluorobenzyl)-2-phenyl-2,3 -dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxo ethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl [Chemical formula] The labeled compound of Step 6 (150 mg, yield 57%) was prepared in the same manner as described in Step 18 of Example 1 from (S)-2-chloro-1-(7-(4-fluorobenzyl)- 2-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 1-yl)ethan-1-one and (2R,5S)-5-(((3R,5R)-3,5-di methylmorpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl LC-MS (M+H) + = 688.4.

[0336] Step 7: 2-((2R,5R)-2-(((3R,5R)-3,5-dimethylmorpho (R)-methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro benzyl)-2-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazin-1-yl)ethan-1-one (Compound 44) Compound 44 (51 mg, yield 66%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino)meth yl)-4-(2-((S)-7-(4-fluorobenzyl)-2-phenyl-2,3- dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxo ethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. 1 H NMR (300 MHz, DMSO-d6) δ 8.62 (s, 1H), 7.82 (d, J = 2.1Hz, 1H), 7.39-7.27 (m, 3H), 7.24 -7.11 (m, 4H), 7.08-6.94 (m,2H), 6.01 (s, 1 H), 4.86 (d, J = 11.7, 1.9Hz, 1H), 4.57-4.4 7 (m, 1H), 4.29 (d, J = 14.4Hz, 1H), 3.92 (s , 2H), 3.55 (dd, J = 11.0,2.9 Hz, 2H), 3.31 (d, J = 11.6 Hz, 1H), 3.25-3.14(m, 3H), 2.9 6 (d, J = 9.0 Hz, 1H), 2.82(dd, J = 11.9, 2.9 Hz, 1H), 2.74-2.65(m, 1H), 2.69-2.63 (m, 3 H), 2.63-2.44 (m, 2H), 2.37(t, J = 11.3 Hz, 1H), 2.07 - 1.95 (m, 1H), 1.18 (d, J = 6.3 Hz, 3H), 0.78 (d, J = 6.3 Hz, 6H). LC - MS (M + H) + = 588.3。

[0337] Example 45: 1 - ((S) - 7 - (4 - fluorobenzyl) - 2 - phenyl - 2,3 - di hydro - 1H - pyrido[2,3 - b][1,4]oxazin - 1 - yl) - 2 - ((2R ,5R) - 5 - methyl - 2 - (((R) - 3 - methylmorpholino)methyl)piperazin - 1 - yl)ethan - 1 - one (Compound 45)

Chemical Structure

Chemical Structure

[0338] Step 2: 1-((S)-7-(4-Fluorobenzyl)-2-phenyl-2,3-di hydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-((2R ,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin- 1-yl)ethan-1-one (Compound 45) Compound 45 (44 mg, yield 35%) as the free base was used in Step 7 of Example 29 in a manner similar to the method described, to prepare from (2R,5S)-4-(2-((S)-7-(4-fluoro benzyl)-2-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1,4] oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methyl morpholino)methyl)piperazine-1-carboxylic acid tert-butyl. 1 H NMR (300 MHz, chloroform-d) δ8.63 (s, 1H), 7.8 0 (s, 1H), 7.35 - 7.11 (m, 7H),7.06 - 6.95 (m, 2H), 6.23 (s, 1H), 4.94 (d,J = 11.5, 1.9 Hz, 1H), 4.54(d, J = 11.6, 3.0 Hz, 2H), 3.92 (s , 2H), 3.60 (d, J = 11.5 Hz,1H), 3.54 - 3.47 ( m, 1H), 3.51 - 3.42 (m, 1H),3.18 - 3.10 (m, 1H ), 3.08 - 2.99 (m, 1H), 2.98(dd, J = 11.4, 2. 3 Hz, 1H), 2.89 - 2.76 (m, 2H),2.76 - 2.67 (m, 2H), 2.55 (t, J = 10.9 Hz,1H), 2.50 (s, 1H), 2.22 - 2.17 (m, 1H), 2.09 (t, J = 10.9 Hz, 1H) , 2.01 - 1.91 (m, 1H), 1.83 (dd, J = 13.4, 3.1 Hz, 1H), 1.04 (d, J = 6.2 Hz, 3H), 0.58 (d, J = 6.3 Hz, 3H). LC-MS (M+H) + = 574.3。

[0339] Example 46: 2 - ((2R,5R)-2 - (((3R,5R)-3,5 - dimethylmorpho olino)methyl)-5 - methylpiperazin - 1 - yl)-1 - ((S)-7-(4 - fluoro benzyl)-2 - isopropyl - 2,3 - dihydro - 1H - pyrido[2,3 - b][1 ,4]oxazin - 1 - yl)ethan - 1 - one (Compound 46)

Chemical formula

Chemical formula

[0340] Step 2: (S)-7-(4 - fluorobenzyl)-2 - isopropyl - 2,3 - dihydro tert-Butyl 1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate butyl

Chem.

[0341] Step 3: (S)-7-(4-Fluorobenzyl)-2-isopropyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazine

Chem.

[0342] Step 4: (S)-2-Chloro-1-(7-(4-fluorobenzyl)-2-isopropyl -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )ethan-1-one

Chem.

[0343] Step 5: (2R,5S)-5-(((3R,5R)-3,5-Dimethylmorpholino) methyl)-4-(2-((S)-7-(4-fluorobenzyl)-2-isopropyl-2 ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2- oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0344] Step 6: 2-((2R,5R)-2-(((3R,5R)-3,5-Dimethylmorph olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro (2R,5S)-5-(((3R,5R)-3,5-Dimethylmorpholino)methyl)-4-(2-((S)-7-(4-fluorobenzyl)-2-isopropyl-2, ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 46) Compound 46 (30 mg, yield 31%) was prepared from (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino)methyl)-4-(2-((S)-7-(4-fluorobenzyl)-2-isopropyl-2, ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl in the same manner as described in Step 7 of Example 29. 1 H NMR (300 MHz, chloroform-d) δ 8.8.26 (s, 1H), 7.83 (s, 1H), 7.19 - 7.08 (m, 2H), 7.03 - 6.92 (m, 2H), 4.66 (d, J = 11.2 Hz, 1H), 4.38 (s, 1H), 4.25 - 4.15 (m, 1H), 3.90 (s, 2H), 3.66 - 3.57 (m, 2H), 3.33 - 3.27 (m, 2H), 3.21 (s, 1H ), 2.78 (s, 4H), 2.72 - 2.62 (m, 1H), 2.58 - 2. 24 (m, 2H), 2.15 - 2.02 (m, 1H), 2.02 - 1.89 (m , 4H), 1.06 (d, J = 6.4 Hz, 3H), 1.03 - 0.84 (m , 12H). LC-MS (M+H) + = 554.4.

[0345] Example 47: 1-((S)-7-(4-Fluorobenzyl)-2-isopropyl-2,3 -dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-(( (2R,5R)-5-Methyl-2-(((R)-3-methylmorpholino)methyl)piperazin -1-yl )ethan-1-one (Compound 47)

Chem.

Chem.

[0346] Step 2: 1-((S)-7-(4-Fluorobenzyl)-2-isopropyl-2,3 -dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-(( (2R,5R)-5-methyl-2-(((R)-3-methylmorpholino)methyl)piperazin -1-yl)ethan-1-one (Compound 47) Compound 47 (36 mg, yield 44%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-2- isopropyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino )methyl)piperazine-1-carboxylic acid tert-butyl. 1 HNMR ( 300 MHz, chloroform-d) δ 8.26 (s,1H), 7.87-7.80 (m, 1H), 7.19-7.08(m, 2H), 7.05-6.92 (m, 2 H), 4.65 (dd, J = 11.4, 1.4Hz, 1H), 4.59-4. 28 (m, 2H), 4.27-4.16 (m, 1H),3.90 (s, 2H), 3.75-3.49(m, 3H), 3.26-2.96 (m, 3H), 2.90 -2.63 (m, 4H), 2.60-2.40 (m,2H), 2.33 (s, 1 H), 2.15 (s, 1H), 2.05-1.76(m, 4H), 1.02 (d d, J = 19.5, 6.4 Hz, 6H), 0.90(s, 6H). LC-MS (M+H) + = 540.5.

[0347] Example 48: 2-((2R,5R)-2-((3,3-dimethylmorpholino)methyl)- 5-methylpiperazin-1-yl)-1-((S)-7-(4-fluorobenzyl)-6 -(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)ethan-1-one (Compound 48) [Chemistry] Step 1: (2R,5S)-4-Benzyl-5-((3,3-dimethylmorpholino)meth yl)-2-methylpiperazine-1-carboxylic acid tert-butyl [Chemistry] The labeled compound of Step 1 (500 mg, yield 81%) was prepared from 3,3-dimethylmorpholine and (2R,5R)-4-benz yl-5-(chloromethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl in the same manner as described in Step 16 of Example 1. LC-MS (M+H) = 418.4. +

[0348] Step 2: (2R,5S)-5-((3,3-dimethylmorpholino)methyl)-2-me thyl [Chemistry] The labeled compound of Step 2 (350 mg, crude) was prepared from (2R,5S)-4-benzyl-5-((3,3-dimethylmorpholi no)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl in the same manner as described in Step 17 of Example 1. LC-MS (M+H) + = 328.3.

[0349] Step 3: (2R,5S)-4-(2-((S)-6-(acetoxymethyl)-7-( 4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-5-((3,3-dimethylmorpho tert-Butyl (S)-1-(2-((2R,5R)-2-((3,3-dimethylmorpholino)methyl)-5-methylpiperazin-1-yl)acetyl)-7-(4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-6-carboxylate

Chem.

[0350] Step 4: ((S)-1-(2-((2R,5R)-2-((3,3-dimethylmorph olino)methyl)-5-methylpiperazin-1-yl)acetyl)-7-(4-fluoroben zyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]ox azine-6-yl)methyl acetate

Chem.

[0351] Step 5: 2-((2R,5R)-2-((3,3-dimethylmorpholino)methyl)- 5-methylpiperazin-1-yl)-1-((S)-7-(4-fluorobenzyl)-6 -(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)ethan-1-one (Compound 48) ((S)-1-(2-((2R,5R)-2-((3,3-dimethylmorpholino)methyl yl)-5-methylpiperazin-1-yl)acetyl)-7-(4-fluorobenzyl)- 2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6 -yl)methyl acetate (120 mg, 0.2 mmol) in THF (5 mL) was added with a solution of LiOH (10 mg, 0.4 mmol) in H2O (5 mL). The resulting mixture was stirred at room temperature for 1 hour. When the reaction was complete, the resulting mixture was concentrated under reduced pressure The residue was purified by preparative HPLC under the following conditions: column XBridge Shield RP1 8 OBD column, 30×150 mm, 5um; mobile phase acetonitrile / water (10 mmol / L NH4HCO3 and 0.1% NH3·H2O), gradient of 23% - 53% in 8 minutes; detector UV 254 nm to give the title compound (47 mg, yield 42%). 1 H NMR (300 MHz, chloroform-d) δ 8. 39 (brs, 1 H), 7.14 - 7.03 (m,2 H), 7.03 - 6.91 (m, 2 H),4.95 (s, 1 H), 4.64 - 4.47 (m, 2 H), ​4.45 - 4.35 (m, 1 H), 4.35 - 4.24 (m, 1 H), 4.19 - 4.08 (m, 1 H), 3.82 (s, 2H), 3.68 - 3.60 (m, 2 H), 3.33 - 3.07 (m, 4 H), 2.89 - 2.76 (m, 1 H) , 2.76 - 2.67 (m, 1 H), 2.57 - 2.40 (m, 4 H), 2. 44 - 2.36 (m, 1 H), 2.23 - 2.01 (m, 2 H), 1.36 - 1 .28 (m, 3 H), 1.05 - 0.98 (m, 3 H), 0.98 - 0.89 (m, 6 H). LC - MS (M + H) + = 556.3。

[0352] Example 49: 2 - ((2R,5R) - 2 - (((2S,5R) - 2,5 - dimethylmorph olino)methyl) - 5 - methylpiperazin - 1 - yl) - 1 - ((S) - 7 - (4 - fluoro benzyl) - 6 - (hydroxymethyl) - 2 - methyl - 2,3 - dihydro - 1H - pyr ido[2,3 - b][1,4]oxazin - 1 - yl)ethan - 1 - one (Compound 49)

Chemical Structure

Chemical Structure

[0353] Step 2: (2R,5S)-5-(((2S,5R)-2,5-Dimethylmorpholino) methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chem.

[0354] Step 3: (2R,5S)-4-(2-((S)-6-(Acetoxymethyl)-7-( 4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-5-(((2S,5R)-2 ,5-Dimethylmorpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert -butyl

Chem.

[0355] Step 4: (2R,5S)-5-(((2S,5R)-2,5-dimethylmorpholino) methyl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl yl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxaz in-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert -butyl

Chemical Structure

[0356] Step 5: 2-((2R,5R)-2-(((2S,5R)-2,5-dimethylmorpho lino)methyl)-5-methylpiperazine-1-yl)-1-((S)-7-(4-fluoro robenzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido (2R,5S)-5-(((2S,5R)-2,5-Dimethylmorpholino)methyl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl (Compound 49) Compound 49 as a formate salt was prepared in a manner similar to the method described in Step 19 of Example 1 (2R,5S)-5-(((2S,5R)-2,5-Dimethylmorpholino)methyl)-4 -(2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl )-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ). The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dried, filtered, and concentrated. The residue was further lyophilized to obtain Compound 49 (15 mg, yield 14%) as the free base. The above salt was neutralized with an aqueous solution of NaHCO3 and extracted with EtOAc. The combined organic layers were dried, filtered, and concentrated. The combined organic layers were dried, filtered, and concentrated. The residue was further lyophilized to obtain Compound 49 (15 mg, yield 14%) as the free base. The residue was further lyophilized to obtain Compound 49 (15 mg, yield 14%) as the free base. 1 1H NMR (400 MHz, DMSO-d 6) δ 8.22 (s, 1H), 7.21 - 7.17(m, 2H), 7.14 - 7.09 (m, 2H), 5.07 (t, J = 5.6 Hz, 1H), 4.7 9 (s, 1H), 4.43 - 4.32 (m,3H), 4.22 - 4.20 ( m, 1H), 4.06 - 4.00 (m, 2H),3.95 - 3.91 (m, 1H), 3.57 - 3.55 (m, 1H), 3.50 - 3.47 (m, 1H) , 3.39 - 3.37 (m, 1H), 3.01 - 2.96 (m, 1H), 2 .89 - 2.86 (m, 2H), 2.77 - 2.74 (m, 1H), 2.71 (brs, 2H),2.60 (brs, 1H), 2.55 (brs, 1H), 2.32 - 2.27 (m, 1H), 2.21 - 2.18 (m, 1H), 1.9 6 (brs, 1H), 1.64 - 1.55 (m,2H), 1.16 (d, J = 6.4 Hz, 3H), 0.97 (d, J =6.2 Hz, 3H), 0.86 (d, J = 6.2 Hz, 6H). LC-MS (M+H) + = 556.5。

[0357] Example 50: 2-((2R,5R)-2-((7-oxa-4-azaspiro[2.5]octan -4-yl)methyl)-5-methylpiperazin-1-yl)-1-((S)-7- (4-fluorobenzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 50)

Chemical Structure

Chemical Structure

[0358] Step 2: (2R,5S)-5-((7-oxa-4-azaspiro[2.5]octane tert-Butyl 2-methylpiperazine-1-carboxylate [ka] The title compound of Step 2 (320 mg, crude) was purified by the method described in Step 17 of Example 1. In a similar manner to the above, (2R,5S)-5-((7-oxa-4-azaspiro[2.5]o (4-methyl-4-ctan-4-yl)methyl)-4-benzyl-2-methylpiperazine-1-carboxylic acid Prepared from ert-butyl. LC-MS (M+H) + = 326.2.

[0359] Step 3: (2R,5S)-5-((7-oxa-4-azaspiro[2.5]octane -4-yl)methyl)-4-(2-((S)-6-(acetoxymethyl)-7-(4-fluorophenyl)- (fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1, 4]Oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carbo tert-Butyl phosphate [ka] The title compound of Step 3 (448 mg, 53% yield for two steps) was synthesized by the same procedure as in Example 1. (S)-(1-(2-chloroacetyl)-7-(2-chlorophenyl)-2-(2-chlorophenyl)-1,2-dichlorophenyl ... (4-Fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b [1,4]oxazin-6-yl)methyl acetate and (2R,5S)-5-((7 -Oxa-4-azaspiro[2.5]octan-4-yl)methyl)-2-methylpipera Prepared from tert-butyl sulphonate-1-carboxylate. LC-MS (M+H) + = 6 96.4.

[0360] Step 4: (2R,5S)-5-((7-oxa-4-azaspiro[2.5]octan -4-yl)methyl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(h ydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1, 4]oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carbo nic acid tert-butyl

Chemical formula

[0361] Step 5: 2-((2R,5R)-2-((7-oxa-4-azaspiro[2.5]o ctan-4-yl)methyl)-5-methylpiperazin-1-yl)-1-((S)-7- (4-fluorobenzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 50) Compound 50 (25 mg, yield 11%) was prepared in the same manner as described in Step 7 of Example 29 ​​by the method of (2R,5S)-5-((7-oxa-4-azaspiro[2.5]octane- 4-yl)methyl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydro xymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carbox ylic acid tert-butyl was prepared. 1 HNMR (300 MHz, chloroform-d) δ 8.40 (brs, 1 H), 7.15 - 7.04(m, 2 H), 7.04 - 6.92 (m, 2 H), 4.91 (s, 1 H),4.66 - 4.47 (m, 2 H), 4.44 - 4.24(m, 2 H), 4.18 - 4.07 (m, 1 H), 3.83 (s, 2 H), 3.69 - 3.63 (m,2H), 3.43 - 3.30 (m, 2 H), 3.20 - 2.98 (m, 2 H),2.89 - 2.76 (m, 4 H), 2.76 - 2.66 (m, 1 H), 2.55 - 2.34(m, 3 H) , 2.18 - 2.05 (m, 1 H), 1.36 - 1.27(m, 3 H), 1. 05 - 0.97 (m, 3 H), 0.64 - 0.41(m, 4 H). LC-MS ( (M + H) + = 554.4.

[0362] Example 51: 2-((2R,5R)-2-(((R)-3-ethylmorpholino)methyl) -5-methylpiperazin-1-yl)-1-((S)-7-(4-fluorobenzyl)- 6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b [1,4]oxazin-1-yl)ethan-1-one (Compound 51) [Chemical formula] Step 1: (2R,5S)-4-Benzyl-5-(((R)-3-ethylmorpholino) methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl [Chemical formula] The labeled compound of Step 1 (550 mg, yield 89%) was prepared in the same manner as described in Step 16 of Example 1 from (R)-3-ethylmorpholine and (2R,5R)-4-benzyl-5-(chloromethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. LC-MS (M+H) = 418.3. +

[0363] Step 2: (2R,5S)-5-(((R)-3-ethylmorpholino)methyl)-2- methylpiperazine-1-carboxylic acid tert-butyl [Chemical formula] The labeled compound of Step 2 (350 mg, crude) was prepared in the same manner as described in Step 17 of Example 1 from (2R,5S)-4-benzyl-5-(((R)-3-ethylmorpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. LC-MS (M+H) + = 328.3.

[0364] Step 3: (2R,5S)-4-(2-((S)-6-(acetoxymethyl)-7-( 4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-5-(((R)-3-ethyl​​​​​​ tert-Butyl (2R,5S)-5-(((R)-3-ethylmorpholino)methyl)-2-methylpiperazine-1-carboxylate [Chemical formula] The title compound of Step 3 (534 mg, yield 58% over 2 steps) was prepared in the same manner as described in Step 18 of Example 1 from (2R,5S)-5-(((R)-3-ethyl morpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl and (( S)-(1-(2-chloroacetyl)-7-(4-fluorobenzyl)-2-methyl-2 ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6-yl)methyl acetate. LC-MS (M+H) + = 698.4.

[0365] Step 4: (2R,5S)-5-(((R)-3-ethylmorpholino)methyl)-4- (2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl)-2-methyl -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl) -2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl [Chemical formula] The title compound of Step 4 (250 mg, yield 88%) was prepared in the same manner as described in Step 4 of Example 8 from (2R,5S)-4-(2-((S)-6-(acetoxymethyl )-7-(4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2 ,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-5-(((R)- ​​tert-Butyl 2-((2R,5R)-2-(((R)-3-ethylmorpholino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one Prepared from tert-butyl . LC-MS (M+H) + = 656.4

[0366] Step 5: 2-((2R,5R)-2-(((R)-3-ethylmorpholino)methyl) -5-methylpiperazin-1-yl)-1-((S)-7-(4-fluorobenzyl)- 6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b [1,4]oxazin-1-yl)ethan-1-one (Compound 51) Compound 51 (52 mg, 24% yield) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-5-(((R)-3-ethylmorpholino)methyl)-4-( 2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl)-2-methyl -2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)- 2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl . 1 H NMR (400 MHz, chloroform-d) δ 8.49 (brs, 1 H), 7.16 - 7.08 (m, 2 H), 7.05 - 6.95 (m, 2 H), 5.01 (s, 1 H), 4.67 - 4.52 (m, 2 H), 4.52 - 4.36 (m, 2 H), 4.36 - 4.26 (m, 1 H), 3.85 (s, 2 H), 3.79 - 3.65 (m, 2 H), 3.59 - 3.50 (m, 1 H), 3.37 -3.21 (m, 2 H), 3.05 - 2.99 (m, 1 H), 2.98 - 2.8 9 (m, 1 H), 2.88 - 2.83 (m, 2 H), 2.79 - 2.70 (m , 1 H), 2.68 - 2.54 (m, 2 H), 2.21 - 2.12 (m, 3 H ), 2.02 - 1.94 (m, 1 H), 1.70 - 1.54 (m, 1 H), 1 .36 - 1.29 (m, 4 H), 1.07 - 1.00 (m, 3 H), 0.92 - 0.84 (m, 3 H). LC - MS (M + H) + = 556.4。

[0367] Example 52: 1 - ((S) - 7 - (4 - fluorobenzyl) - 6 - (hydroxymethyl) - 2 - methyl - 2,3 - dihydro - 1H - pyrido[2,3 - b][1,4]oxazin - 1 - yl) - 2 - ((2R,5R) - 2 - (((R) - 3 - (methoxymethyl)morpho olino)methyl) - 5 - methylpiperazin - 1 - yl)ethan - 1 - one (Compound 52)

Chemical formula

Chemical formula

[0368] Step 2: (2R,5S) - 5 - (((R) - 3 - (methoxymethyl)morpholino)me tert-Butyl (2R,5S)-4-benzyl-5-(((R)-3-(methoxymethyl)morpholino)methyl)-2-methylpiperazine-1-carboxylate

Chem.

[0369] Step 3: (2R,5S)-4-(2-((S)-6-(acetoxymethyl)-7-( 4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-5-(((R)-3-(meth oxymethyl)morpholino)methyl)-2-methylpiperazine-1-carboxylate tert- butyl

Chem.

[0370] ​ Step 4: (2R,5S)-4-(2-((S)-7-(4-Fluorobenzyl)-6 -(Hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-5-(((R)-3-(meth oxymethyl)morpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert- butyl

Chemical Structure

[0371] Step 5: 1-((S)-7-(4-Fluorobenzyl)-6-(hydroxymethyl) -2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine- 1-yl)-2-((2R,5R)-2-(((R)-3-(methoxymethyl)morpho olino)methyl)-5-methylpiperazin-1-yl)ethan-1-one (Compound 52) Compound 52 (45 mg, yield 26%) as the free base was prepared in the same manner as described in Step 7 of Example 29 to give (2R,5S)-4-(2-((S)-7-(4-Fluoro ​​(Benzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-5-(((R )-3-(methoxymethyl)morpholino)methyl)-2-methylpiperazine-1-carbo nic acid tert-butyl was prepared from 1 HNMR (300 MHz, chloroform-d ) δ 8.41 (brs, 1 H), 7.17 - 7.06(m, 2 H), 7.06 -6.94 (m, 2 H), 4.97 (s, 1H), 4.68 - 4.46 (m, 3 H), 4.45 - 4.27 (m, 2 H), 3.86(s, 2 H), 3.77 -3.60 (m, 2 H), 3.58 - 3.42 (m,3 H), 3.39 - 3.3 3 (m, 2 H), 3.26 (s, 3 H),3.14 - 2.83 (m, 4 H) , 2.83 - 2.51 (m, 3 H), 2.41(s, 1 H), 2.32 - 1. 91 (m, 3 H), 1.36 - 1.28 (m,3 H), 1.07 - 0.99 ( m, 3 H). LC-MS (M + H) + = 572.4

[0372] Example 53: 2-((2R,5R)-2-(((R)-3-(ethoxymethyl)morpho olino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro benzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 53)

Chemical Structure

Chem.

[0373] Step 2: (R)-3-(Ethoxymethyl)morpholine

Chem.

[0374] Step 3: (2R,5S)-4-Benzyl-5-(((R)-3-(ethoxymethyl) morpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chem.

[0375] Step 4: (2R,5S)-5-(((R)-3-(ethoxymethyl)morpholino)me thyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chem.

[0376] Step 5: (2R,5S)-4-(2-((S)-6-(acetoxymethyl)-7-( 4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-5-(((R)-3-(eth oxymethyl)morpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert- butyl

Chem.

[0377] Step 6: (2R,5S)-5-(((R)-3-(ethoxymethyl)morpholino)me thyl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl ))-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert- butyl

Chem.

[0378] Step 7: 2-((2R,5R)-2-(((R)-3-(ethoxymethyl)morpho no)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro benzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 53) Compound 53 (54 mg, yield 42%) was prepared in the same way as described in Step 7 of Example 29 from (2R,5S)-5-(((R)-3-(ethoxymethyl)morpholino)methy l)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl) -2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-but yl. 1 H NMR(400 MHz, DMSO-d6) δ 8.19 ( brs, 1 H), 7.26-7.17 (m, 2H), 7.17-7.07 (m, 2 H), 5.12-5.04(m, 1 H), 4.75 (s, 1 H), 4.4 9-4.30 (m, 3 H), 4.25-4.17(m, 1 H), 4.02-3. 93 (m, 3 H), 3.68-3.53 (m,2 H), 3.46-3.33 ( m, 4 H), 3.32-3.23 (m, 2 H),2.90-2.81 (m, 1 H), 2.81-2.73 (m, 3 H), 2.66-2.53(m, 2 H), 2.34-2.18 (m, 3 H), 2.08-1.98(m, 1 H), 1.96 -1.87 (m, 1 H), 1.21-1.15 (m,3 H), 1.10-1.0 2 (m, 3H), 0.89-0.83 (m, 3H). LC-MS (M+H) + = 586.4.

[0379] Example 54: 1-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl) -2-Methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine- 1-yl)-2-((2R,5R)-2-(((R)-3-(hydroxymethyl)morpho (5-methyl-5-piperazin-1-yl)ethan-1-one (compound 54) [ka] Step 1: (S)-4-benzyl-3-(((tert-butyldimethylsilyl)oxy) Methyl)morpholine [ka] [(3R)-4-benzylmorpholin-3-yl]methanol (425 mg, 2.1 m mol) and imidazole (350 mg, 5.2 mmol) in DMF (10 mL) Add TBDMSCl (375 mg, 2.5 mmol) in small portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 16 hours. The reaction mixture was quenched with H2O at room temperature. The resulting mixture was diluted with EtOAc ( It was extracted with (2×30 mL). The organic phases were combined, washed with saturated brine, and dehydrated over Na2SO4 and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (P E:EtOAc = 4:1) to obtain the title compound (342 mg, yield 76 %). LC-MS (M+H) + = 322.2

[0380] Step 2: (S)-3-(((tert-butyldimethylsilyl)oxy)methyl)morpholine

Chem.

Chem.

[0381] Step 3: (2R,5S)-4-benzyl-5-(((S)-3-(((tert-butyl dimethylsilyl)oxy)methyl)morpholino)methyl)-2-methylpiperazine- 1-carboxylic acid tert-butyl

Chem.

[0382] Step 4: (2R,5S)-5-(((S)-3-(((tert-butyldimethylsilyl oxy)methyl)morpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0383] Step 5: (2R,5S)-4-(2-((S)-6-(acetoxymethyl)-7-( 4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-5-(((S)-3-((( tert-butyldimethylsilyl)oxy)methyl)morpholino)methyl)-2-methyl piperazine-1-carboxylic acid tert-butyl

Chemical formula

[0384] Step 6: (2R,5S)-5-(((S)-3-(((tert-butyldimethylsily l)oxy)methyl)morpholino)methyl)-4-(2-((S)-7-(4-fluoro benzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl tert-butyl piperazine-1-carboxylate

Chemical formula

[0385] Step 7: 1-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl) -2-Methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 1-yl)-2-((2R,5R)-2-(((R)-3-(hydroxymethyl)morpho lino)methyl)-5-methylpiperazin-1-yl)ethan-1-one (Compound 54) Compound 54 (70 mg, yield 59%) was prepared in the same way as described in Step 7 of Example 29 from (2R,5S)-5-(((S)-3-(((tert-butyldimethylsily l)oxy)methyl)morpholino)methyl)-4-(2-((S)-7-(4-fluoro benzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methylpi perazine-1-carboxylic acid tert-butyl. 1 HNMR (400 MHz, chloroform-d) δ 8.43 (brs, 1H), 7.16 - 7.07 (m, 2H), 7 .05 - 6.96 (m, 2H), 4.88 (s, 1H), 4.67 - 4.51 (m, 2H ), 4.49 - 4.29 (m, 3H), 3.96 - 3.67 (m, 6H), 3.65 - 3 .48 (m, 3H), 3.30 - 3.22 (m, 1H), 3.13 - 3.00 (m, 2H ), 2.99 - 2.86 (m, 2H), 2.84 - 2.77 (m, 1H), 2.67 - 2 .63 (m, 2H), 2.40 - 2.35 (m, 1H), 2.31 - 2.21 (m, 1H ), 2.18 - 2.08 (m, 2H), 1.35 - 1.29 (m, 3H), 1.08 - 1 .02 (m, 3H). LC-MS (M + H) + = 558.3.

[0386] Example 55: 2-((2R,5R)-2-(((S)-3-(difluoromethyl)morpho (R)-Methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro benzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyr ido[2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 55)

Chemical Structure

Chemical Structure

[0387] Step 2: (S)-4-Benzyl-3-(difluoromethyl)morpholine

Chemical formula

[0388] Step 3: (S)-3-(Difluoromethyl)morpholine hydrochloride

Chemical formula

[0389] Step 4: (2R,5S)-4-Benzyl-5-(((S)-3-(difluoromethyl )morpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0390] Step 5: (2R,5S)-5-(((S)-3-(difluoromethyl)morpholino) methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0391] Step 6: (2R,5S)-4-(2-((S)-6-(acetoxymethyl)-7-( 4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b] [1,4]oxazin-1-yl)-2-oxoethyl)-5-(((S)-3-(dif luoromethyl)morpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert -butyl

Chem.

[0392] Step 7: (2R,5S)-5-(((S)-3-(difluoromethyl)morpholino) methyl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl yl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxaz in-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert -butyl

Chem.

[0393] Step 8: 2-((2R,5R)-2-(((S)-3-(difluoromethyl)morpho lino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-fluoro benzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 55) Compound 55 (53 mg, yield 46%) was prepared from tert-butyl (2R,5S)-5-(((S)-3-(difluoromethyl)morpholino)meth yl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl )-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid in the same manner as described in Step 7 of Example 29. 1 H NMR (400 MHz, chloroform-d) δ 8.41 (b rs, 1H), 7.14 - 7.04 (m, 2H), 7.03 - 6.92 (m, 2H), 6 .32 - 5.87 (m, 1H), 5.06 - 4.77 (m, 1H), 4.64 - 4.50 (m, 2H), 4.44 - 4.35 (m, 1H), 4.35 - 4.25 (m, 1H), 4 .10 - 4.02 (m, 1H), 3.83 (s, 2H), 3.82 - 3.57 (m, 4H ), 3.23 - 3.07 (m, 2H), 2.95 - 2.79 (m, 3H), 2.78 - 2 .64 (m, 2H), 2.62 - 2.36 (m, 4H), 2.19 - 2.07 (m, 1H ), 1.35 - 1.23 (m, 3H), 1.06 - 0.99 (m, 3H). LC - MS (M + H) + = 578.4。

[0394] Example 56: 2 - ((2R,5R) - 2 - ((8 - oxa - 5 - azaspiro[3.5]non an - 5 - yl)methyl) - 5 - methylpiperazin - 1 - yl) - 1 - ((S) - 7 - ( 4 - fluorobenzyl) - 6 - (hydroxymethyl) - 2 - methyl - 2,3 - dihydro - 1H - pyrido[2,3 - b][1,4]oxazin - 1 - yl)ethan - 1 - one (Compound 56)

Chemical Structure

Chemical Structure

[0395] Step 2: (2R,5S)-5-((8-Oxa-5-azaspiro[3.5]nonan- 5-yl)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0396] Step 3: (2R,5S)-5-((8-Oxa-5-azaspiro[3.5]nonan- 5-yl)methyl)-4-(2-((S)-6-(acetoxymethyl)-7-(4-flu orobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazine-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

[0397] Step 4: (2R,5S)-5-((8-Oxa-5-azaspiro[3.5]nonan- 5-yl)methyl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydro xymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl [Chemical formula] The title compound of Step 4 (250 mg, yield 75%) was prepared in the same manner as described in Step 4 of Example 8 from (2R,5S)-5-((8-oxa-5-azaspiro[3.5 nonan-5-yl)methyl)-4-(2-((S)-6-(acetoxymethyl)-7- (4-fluorobenzyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b (1,4]oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1 -carboxylic acid tert-butyl. LC-MS (M+H) = 668.4 + .

[0398] Step 5: 2-((2R,5R)-2-((8-Oxa-5-azaspiro[3.5]no nan-5-yl)methyl)-5-methylpiperazine-1-yl)-1-((S)-7-( 4-fluorobenzyl)-6-(hydroxymethyl)-2-methyl-2,3-dihydro- ​(1H-Pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 56) Compound 56 (59 mg, yield 28%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-5-((8-oxa-5-azaspiro[3.5]nonan-5 -yl)methyl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydro xymethyl)-2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4] oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. 1 1H NMR (300 MHz, chloroform-d) δ 8. 45 (brs, 1H), 7.17 - 7.06 (m, 2H), 7.06 - 6.93 (m, 2 H), 4.97 (s, 1H), 4.66 - 4.50 (m, 2H), 4.47 - 4.37 ( m, 1H), 4.37 - 4.24 (m, 2H), 3.85 (s, 2H), 3.72 - 3. 56 (m, 3H), 3.48 - 3.38 (m, 1H), 3.22 - 3.08 (m, 2H) , 2.93 - 2.66 (m, 3H), 2.63 - 2.32 (m, 4H), 2.32 - 2. 20 (m, 1H), 2.17 - 2.00 (m, 3H), 1.87 - 1.49 (m, 4H) , 1.37 - 1.29 (m, 3H), 1.07 - 0.99 (m, 3H). LC-MS ( M + H) + = 568.3.

[0399] Example 57: 1-((S)-7-(4-Fluorobenzyl)-6-(hydroxymethyl) -2-(methoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4] oxazin-1-yl)-2-((2R,5R)-5-methyl-2-(((R)-3-meth ​((2R)-2-(morpholin-4-ylmethyl)piperazin-1-yl)ethan-1-one (Compound 57) [Chemical formula] Step 1: (S)-(1-(Benzyloxy)-3-((3,5-dibromo-6-methyl pyridin-2-yl)oxy)propan-2-yl)carbamic acid tert-butyl [Chemical formula] The titled compound of Step 1 (9.8 g, yield 99%) was prepared from (R)-(1-(benzyloxy)-3-hydroxypropan- 2-yl)carbamic acid tert-butyl and 3,5-dibromo-2-chloro-6-methyl pyridine in the same manner as described in Step 1 of Example 19. LC-MS (M+H) = 529.0. +

[0400] Step 2: (S)-2-((Benzyloxy)methyl)-7-bromo-6-methyl-2 ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylic acid t ert-butyl [Chemical formula] The titled compound of Step 2 (4 g, yield 74%) was prepared from (S)-(1-(benzyloxy)-3-((3,5-dibromo-6 -methylpyridin-2-yl)oxy)propan-2-yl)carbamic acid tert-but yl in the same manner as described in Step 2 of Example 34. LC-MS (M+H) = 449.1. +

[0401] ​​Step 3: (S)-2-((benzyloxy)methyl)-7-(4-fluorobenzyl )-6-Methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine 1-tert-Butyl carboxylate [ka] The title compound of Step 3 (2.9 g, 91% yield) was synthesized by the procedure described in Step 7 of Example 1. In a similar manner, (S)-2-((benzyloxy)methyl)-7-bromo-6-methyl Chil-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxamide It was prepared from tert-butyl benzoate and (4-fluorobenzyl)zinc(II) chloride. LC-MS (M+H) + = 479.2.

[0402] Step 4: (S)-7-(4-fluorobenzyl)-2-(hydroxymethyl)-6- Methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-carboxylate tert-Butyl carboxylate [ka] The title compound of Step 4 (2 g, 84% yield) was synthesized using the method described in Step 4 of Example 34. In a similar manner to the above, (S)-2-((benzyloxy)methyl)-7-(4-fluorobenzyloxy)methyl Benzyl)-6-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]ox Prepared from tert-butyl sazine-1-carboxylate. LC-MS (M+H) + = 389.2.

[0403] Step 5: (R)-7-(4-fluorobenzyl)-6-methyl-2-((methylsulfonyl) (Rhonyl)oxy)methyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazine-1-carboxylic acid tert-butyl

Chemical formula

[0404] Step 6: (R)-(7-(4-Fluorobenzyl)-6-methyl-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-2-yl)methyl methanesulfonate

Chemical formula

[0405] Step 7: (S)-7-(4-Fluorobenzyl)-2-(methoxymethyl)-6-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine ​​​​​​​ [Chemical formula] The labeled compound of Step 7 (665 mg, yield 58%) was prepared from (R)-(7-(4-fluorobenzyl)-6-methyl-2, 3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-2-yl)methyl meth anesulfonate and sodium methoxide in the same manner as described in Step 7 of Example 34. LC-MS (M+H) = + = 303.1.

[0406] Step 8: (S)-7-(4-Fluorobenzyl)-2-(methoxymethyl)-6-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine-1-car boxylic acid tert-butyl [Chemical formula] The labeled compound of Step 8 (590 mg, yield 79%) was prepared from (S)-7-(4-fluorobenzyl)-2-(methoxymethyl )-6-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine in the same manner as described in Step 4 of Example 1. LC-MS (M+H) = 403.2. + = 403.2.

[0407] Step 9: (S)-1-(tert-Butoxycarbonyl)-7-(4-fluorobenz zyl)-2-(methoxymethyl)-6-methyl-2,3-dihydro-1H-pyrido[2, [Chemical formula] The labeled compound of Step 9 (457 mg, yield 74%) was prepared in the same manner as described in Step 5 of Example 1 from (S)-7-(4-fluorobenzyl)-2-(methoxymethyl )-6-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine -1-carboxylic acid tert-butyl. LC-MS (M+H) + = 419 .2。

[0408] Step 10: (S)-6-(acetoxymethyl)-7-(4-fluorobenzyl)-2 -(methoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]ox azine-1-carboxylic acid tert-butyl

Chemical formula

[0409] Step 11: (S)-(7-(4-fluorobenzyl)-2-(methoxymethyl)-2 ,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-6-yl)methyl acetate

Chemical formula

[0410] Step 12: (S)-(1-(2-Chloroacetyl)-7-(4-fluorobenzyl) -2-(methoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4] oxazin-6-yl)methyl acetate

Chemical formula

[0411] Step 13: (2R,5S)-4-(2-((S)-6-(acetoxymethyl)-7- (4-fluorobenzyl)-2-(methoxymethyl)-2,3-dihydro-1H-pyrido [2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl- 5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert -butyl

Chemical formula

[0412] Step 14: (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl)-2-(methoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl-5-(((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-butyl

Chemical Structure

[0413] ​​​​​​​​​​​​​​Step 15: 1-((S)-7-(4-Fluorobenzyl)-6-(hydroxymethyl )-2-(methoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazin-1-yl)-2-((2R,5R)-5-methyl-2-(((R)-3- methylmorpholino)methyl)piperazin-1-yl)ethan-1-one (Compound 57) Compound 57 (20 mg, yield 15%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-4-(2-((S)-7-(4-fluorobenzyl)-6- (hydroxymethyl)-2-(methoxymethyl)-2,3-dihydro-1H-pyrido[2 ,3-b][1,4]oxazin-1-yl)-2-oxoethyl)-2-methyl-5- (((R)-3-methylmorpholino)methyl)piperazine-1-carboxylic acid tert-but yl. 1 H NMR (300 MHz, DMSO-d6) δ 8.22 ( brs, 1 H), 7.25 - 7.14 (m, 2H), 7.17 - 7.04 (m, 2 H), 5.14 - 5.04(m, 1 H), 4.85 (s, 1 H), 4.5 4 - 4.45 (m, 1 H), 4.45 - 4.33(m, 2 H), 4.25 - 4. 09 (m, 2 H), 3.96 (s, 2 H),3.54 - 3.45 (m, 2 H ), 3.49 - 3.33 (m, 1 H), 3.30 - 3.24(m, 2H), 3. 21 (s, 3 H), 3.07 - 2.95 (m,1 H), 2.93 - 2.74 ( m, 2 H), 2.73 - 2.53 (m, 4H),2.39 - 2.12 (m, 3 H), 2.10 - 1.96 (m, 1 H), 1.82 - 1.71(m, 1 H), 1.22 (s, 1 H), 0.92 - 0.85 (m, 3 H), 0.85 - 0.79 (m, 3H). LC-MS (M + H) + = 572.0。

[0414] Example 58: 2 - ((2R,5R)-2 - (((3R,5R)-3,5 - dimethylmorpho olino)methyl)-5 - methylpiperazin - 1 - yl)-1 - ((S)-7-(4 - fluoro benzyl)-6 - (hydroxymethyl)-2 - (methoxymethyl)-2,3 - dihydro -1H - pyrido[2,3 - b][1,4]oxazin - 1 - yl)ethan - 1 - one (Compound 58)

Chemical Structure

Chemical Structure

[0415] Step 2: (2R,5S)-5-(((3R,5R)-3,5-Dimethylmorpholino) methyl)-4-(2-((S)-7-(4-Fluorobenzyl)-6-(hydroxymethyl yl)-2-(methoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1, 4]oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carbo nic acid tert-butyl

Chemical formula

[0416] Step 3: 2-((2R,5R)-2-(((3R,5R)-3,5-Dimethylmorpho lino)methyl)-5-methylpiperazin-1-yl)-1-((S)-7-(4-Fluoro benzyl)-6-(hydroxymethyl)-2-(methoxymethyl)-2,3-dihydro -1H-pyrido[2,3-b][1,4]oxazin-1-yl)ethan-1-one (Compound 58) ​​Compound 58 (29 mg, yield 32%) was prepared in the same manner as described in Step 7 of Example 29 from (2R,5S)-5-(((3R,5R)-3,5-dimethylmorpholino)meth yl)-4-(2-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl ))-2-(methoxymethyl)-2,3-dihydro-1H-pyrido[2,3-b][1,4 oxazin-1-yl)-2-oxoethyl)-2-methylpiperazine-1-carboxylic acid tert-butyl. 1 1H NMR (400 MHz, DMSO-d6) δ 8.20 (brs, 1 H), 7.25 - 7.17 (m, 2 H), 7.21 - 7.07 (m, 2 H), 5.13 - 5.06 (m, 1 H), 4.86 (s, 1 H), 4.55 - 4.36 (m, 3 H), 4.26 - 4.18 (m, 1 H), 4.09 - 3.95 (m, 3 H), 3.52 - 3.44 (m, 2 H), 3.43 - 3.38 (m, 2 H), 3.24 (s, 3H), 3.19 - 3.11 (m, 2 H), 3.09 - 2.98 (m, 1 H), 2.84 - 2.75 (m, 1 H) , 2.73 - 2.51 (m, 5 H), 2.50 - 2.44 (m, 1 H), 2. 30 - 2.20 (m, 1 H), 2.19 - 2.10 (m, 1 H), 2.00 - 1 .90 (m, 1 H), 0.90 - 0.80 (m, 9 H). LC-MS (M+H) + = 586.3.

[0417] Example 59: 1-((S)-7-(4-fluorobenzyl)-6-(hydroxymethyl) -2-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazine- 1-yl)-2-((2R,5R)-2-(((S)-3-(isopropoxymethyl)mo lholino)methyl)-5-methylpiperazin-1-yl)ethan-1-one (Compound 59 )

Chemical formula

Chemical formula

[0418] Step 2: (S)-4-Benzyl-3-(isopropoxymethyl)morpholine

Chemical formula

[0419] Step 3: (S)-3-(isopropoxymethyl)morpholine

Chemical formula

[0420] Step 4: (2R,5S)-4-benzyl-5-(((S)-3-(isopropoxymeth yl)morpholino)methyl)-2-methylpiperazine-1-carboxylic acid tert-butyl

Chemical formula

Claims

[Claim 1] The invention described in the specification.

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