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WO2025177150A1 - Compound as parp1 inhibitor and pharmaceutical composition comprising the same - Google Patents

Compound as parp1 inhibitor and pharmaceutical composition comprising the same

Info

Publication number
WO2025177150A1
WO2025177150A1 PCT/IB2025/051745 IB2025051745W WO2025177150A1 WO 2025177150 A1 WO2025177150 A1 WO 2025177150A1 IB 2025051745 W IB2025051745 W IB 2025051745W WO 2025177150 A1 WO2025177150 A1 WO 2025177150A1
Authority
WO
WIPO (PCT)
Prior art keywords
methyl
mmol
fluoro
dihydrooxazin
pyridin
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/IB2025/051745
Other languages
French (fr)
Inventor
Moo Sung Ko
Il Hyang Kim
Younghue HAN
Jaeyoung Lee
Dahae Lee
Mijin Yoon
Uji KIM
Keesoo NAM
Junho Cho
Hyunmo YANG
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Chong Kun Dang Corp
Original Assignee
Chong Kun Dang Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Chong Kun Dang Corp filed Critical Chong Kun Dang Corp
Publication of WO2025177150A1 publication Critical patent/WO2025177150A1/en
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5355—Non-condensed oxazines and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/538—1,4-Oxazines, e.g. morpholine ortho- or peri-condensed with carbocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5383—1,4-Oxazines, e.g. morpholine ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5386—1,4-Oxazines, e.g. morpholine spiro-condensed or forming part of bridged ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/5415—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with carbocyclic ring systems, e.g. phenothiazine, chlorpromazine, piroxicam
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/04—Ortho-condensed systems

Definitions

  • the present invention relates to a compound, stereoisomers thereof, pharmaceutically acceptable salts thereof as a PARP1 inhibitor and a pharmaceutical composition including the same, and a method for preventing or treating PARP1 activity- associated diseases using the same and a use thereof.
  • PARP poly(ADP-ribose)polymerase
  • PARP1 and PARP2 are the most extensively studied PARPs for their roles in DNA damage repair.
  • PARP1 is activated by DNA damage break and functions to catalyze the addition of poly(ADP-ribose) (PAR) chains to a target protein.
  • PARylation This post-translational modification, known as PARylation, mediates recruitment of additional DNA repair factors into DNA lesions.
  • PARP self-PARylation triggers release of bound PARP from DNA, thus allowing access to other DNA repair proteins to complete the repair.
  • binding of PARP to an injured site, catalytic activity thereof, and eventual release thereof from DNA are all important steps in which cancer cells respond to DNA damage caused by chemotherapy and radiation therapy (Bai P. Biology of poly(ADP-ribose) polymerases: the factotums of cell maintenance. Mol Cell 2015;58:947-58.).
  • Inhibition of the PARP family enzyme was utilized as a strategy to selectively kill cancer cells by inactivating complementary DNA repair pathways.
  • a number of preclinical and clinical studies have demonstrated that tumor cells, including deleterious alterations of BRCA1 or BRCA2, the major tumor suppressor proteins involved in double-strand DNA break (DSB) repair by homologous recombination (HR), are selectively sensitive to small molecule inhibitors of DNA repair enzymes of the PARP family. These tumors have a defective homologous recombination repair (HRR) pathway and rely on PARP enzyme function for survival.
  • HRR homologous recombination repair
  • Patent Document 0001 International Patent Publication No. WO 2021/013735
  • Patent Document 0002 International Patent Publication No. WO 2021/260092
  • Non-Patent Document 0001 J. Med. Chem. 2021, 64, 19, 14498-14512
  • the present invention may provide a compound represented by a following formula
  • M is CR X , N, NR y , O, or S,
  • Ri and R2 are each independently H, halogen, OH, CN, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, or O-(C1-C5 alkyl),
  • M is CR X , N, O or S
  • X, Y and Z are each independently N or CR Z , each of R x and R z is independently H or halogen,
  • RA and RB are each independently H or C1-C5 alkyl, or RA and RB may be bonded to each other to form three- to seven-membered cycloalkyl,
  • Ri and R2 are each independently H, halogen, C1-C5 alkyl, C1-C5 haloalkyl, or O- (C1-C5 alkyl),
  • A is C6-C12 arylene or five- to 12-membered heteroarylene including one to three heteroatoms independently selected from the group consisting of N, O and S, and halogen may be F.
  • M is CR X , N, O or S
  • X, Y and Z are each independently N or CR Z , each of R x and R z is independently H or halogen,
  • RA and RB are each independently H or C1-C3 alkyl, or RA and RB may be bonded to each other to form four- to six-membered cycloalkyl, is a single bond or a double bond, is absent or a single bond,
  • Ri and R2 are each independently H, halogen, C1-C3 alkyl, C1-C3 haloalkyl, or O- (C1-C3 alkyl),
  • A is phenylene or five- to six-membered heteroarylene including one to three Ns, and halogen may be F.
  • Cm-Cn (in which m and n are each independently an integer of 1 or more) may mean the number of carbons, for example, “C1-C5 alkyl” may represent alkyl having one to five carbon atoms.
  • alkyl may mean a linear or branched saturated hydrocarbon group.
  • alkyl may have one to five carbon atoms.
  • alkyl may have one to four carbon atoms.
  • alkyl may have one to three carbon atoms.
  • Examples of alkyl may include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, n-pentyl, sec -pentyl, tert-pentyl, isopentyl, sec-isopentyl, neo-pentyl, and the like, but are not limited thereto.
  • cycloalkyl may mean a saturated hydrocarbon ring having three or more carbon atoms, which may include both monocyclic and polycyclic structures.
  • cycloalkyl may be a ring having three to 12, three to ten, three to eight, or three to seven carbon atoms.
  • cycloalkyl may include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclopentyl, bicyclohexyl, bicycloheptyl, bicyclooctyl, bicyclononyl, spiropentyl, spirohexyl, spiroheptyl, spirooctyl, spirononyl, and the like, but are not limited thereto.
  • heterocyclo alkyl may mean a cyclic functional group in which at least one or more carbon atoms constituting the above-defined cycloalkyl ring are substituted with a heteroatom.
  • the heteroatom may include nitrogen (N), oxygen (O), or sulfur (S).
  • the heteroatoms included in the ring of heterocycloalkyl may be one type or two or more types, one or one or more of one type of heteroatom may be included therein, and at least one or more of the two or more types of heteroatoms may be included therein, respectively.
  • heterocycloalkyl may be a three- to 12-membered ring.
  • heterocycloalkyl may include oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, morpholinyl, piperazinyl, thiomorpholinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like, but are not limited thereto.
  • aryl may mean a monocyclic or polycyclic aromatic hydrocarbon ring. As one example, in the present invention, aryl may have six to 20 carbon atoms. As one example, in the present invention, aryl may have six to 12 carbon atoms. Examples of aryl may include phenyl, biphenyl, naphthalenyl, and the like, but are not limited thereto.
  • heteroaryl may mean a monocyclic or polycyclic aromatic hydrocarbon ring including at least one or more heteroatoms such as nitrogen (N), oxygen (O), or sulfur (S).
  • heteroaryl may be a five- to 12- membered ring.
  • heteroaryl may be a five- to 12- membered ring having one to three heteroatoms.
  • heteroaryl may be a five- to eight-membered ring having one to three heteroatoms.
  • heteroaryl may be a five- or six-membered ring having one or two heteroatoms.
  • stereoisomer may include a diastereomer and an optical isomer, which may include a single enantiomer, a mixture of enantiomers including a racemate, a single diastereomer, and a mixture of diastereomers all.
  • optical isomer may include a single enantiomer, a mixture of enantiomers including a racemate, a single diastereomer, and a mixture of diastereomers all.
  • Such isomer may be separated by resolution according to conventional techniques, for example, column chromatography, HPLC, or the like.
  • the isomer may be stereospecifically synthesized by using a known array of optically pure starting materials and/or reagents.
  • the isomer may be an optical isomer.
  • “pharmaceutically acceptable salts” may mean salts conventionally used in a pharmaceutical field, and may include, for example, inorganic ion salts prepared from calcium, potassium, sodium, magnesium, and the like; inorganic acid salts prepared from hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, sulfuric acid, hydroiodic acid, etc.; organic acid salts prepared from acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbric acid, carbonic acid, vanillic acid, etc.; sulfonic acid salts prepared from methanesulfonic acid, ethan
  • the compound represented by formula 1 of the present invention may be the same as the compound list in this specification, but also include a pharmaceutically acceptable isotopic-labeled compound in which at least one or more atoms are replaced with an atom having the same atomic number but having an atomic mass or mass number different from the atomic mass or mass number prevailing in nature.
  • isotopes which may be included in the compound of the present invention may include: isotope of hydrogen, 2 H, 3 H; isotope of carbon, n C, 13 C, 14 C; isotope of chlorine, 36 Cl;isotope of fluorine, 18 F; isotope of iodine, 123 I, 125 I; isotope of nitrogen, 13 N, 15 N; isotope of oxygen, 15 0, 17 0, 18 O; isotope of phosphorus, 32 P; isotope of sulfur, 35 S; and the like.
  • a certain isotopic-labeled compound of the present invention may be useful in studying drugs and/or a distribution of substrate tissues (e.g., assays).
  • a radioactive isotope tritium, that is, 3 H, and carbon- 14, that is, 14 C may be useful in view of ease of incorporation and means of immediate detection.
  • Substitution with heavier isotopes for example, substitution of hydrogen ( X H) with deuterium ( 2 H), may exhibit an excellent therapeutic effect on diseases by enhancing metabolic stability, such as increasing a half-life in vivo or reducing a dosage.
  • prevention may refer to all the acts, which inhibit or delay the occurrence of a disease by administering the compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
  • treatment may refer to all the acts, by which a symptom of an individual likely to develop or suffering from a disease gets better or takes a favorable turn by administering the compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
  • the compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof may show an effect of preventing or treating the PARP1 activity-associated diseases at a level similar to, substantially the same as, or higher than that of a conventionally known drug for preventing or treating the PARP1 activity-associated diseases.
  • the present invention may provide a method for preparing the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
  • the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof may be prepared according to any one method of reaction formulas 1 to 4, which may be modified to a level obvious to those skilled in the art.
  • Halo may mean F, Cl, Br or I, and each of Ri, R2, RA, RB, A, X, Y, Z and M may be substantially the same as defined in formula 1, if not specifically defined.
  • a compound of formula 1-1 may be subjected to a substitution reaction with an amine compound to prepare a compound of formula 1-2, which may be subjected to a Suzuki reaction to prepare a compound of formula 1-3, and a protecting group may be removed to prepare a compound of formula 1-4.
  • the compound of formula 1-1 may be subjected to a Suzuki reaction to prepare a compound of formula 2-1, which may be subjected to a substitution reaction with the amine compound to prepare the compound of formula 1-3, and a protecting group may be removed to prepare the compound of formula 1-4.
  • the compound of formula 1-1 may be subjected to a Suzuki reaction to prepare the compound of formula 2-1, which may be subjected to a hydrolysis reaction to prepare a compound of formula 3-1, which may be subjected to a substitution reaction with the amine compound to prepare the compound of formula 1-3, and a protecting group may be removed to prepare the compound of formula 1-4.
  • a compound of formula 4-1 and the compound of formula 1-4 may be subjected to a substitution reaction, or a compound of formula 4-2 and the compound of formula 1-4 may be subjected to a reductive amination reaction to prepare a compound of formula 4-3.
  • the cancer may be at least one selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, hematological cancer, gastrointestinal cancer such as gastric cancer or colorectal cancer, lung cancer such as small cell lung cancer or non-small cell lung cancer, and brain cancer such as glioma or glioblastoma.
  • the inflammatory disease may be at least one selected from the group consisting of rheumatoid arthritis, multiple sclerosis, Crohn's disease, ulcerative colonitis, ulcerative colitis, graft-versus-host disease (GVHD), systemic lupus erythematosus, toxic shock syndrome, osteoarthritis, and insulin-dependent diabetes mellitus.
  • GVHD graft-versus-host disease
  • the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof of the present invention may show an inhibitory effect against PARP1 and may be advantageously used for preventing or treating PARP1 activity-associated diseases.
  • Mode for Invention
  • example compound 48 of 5-[2-[(5-fluoro-3-oxo-spiro[4H-l,4-benzoxazin-2,l'- cyclobutene]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.050 g, 68.46%) in a white solid form.
  • example compound 78 of 5-[2-[(2-ethyl-8-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6- dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2-carboxamide (0.070 g, 55.70%) in a white solid form.
  • example compound 88 of 5-[2-[(3,3-dimethyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7-yl)methyl]-3,6- dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2-carboxamide (0.038 g, 51.71%) in a white solid form.
  • the PARP1/PARP2 binding ability of the test substance was measured using PARPtrapTM Assay Kit for PARP1 (BPS Bio #80584)/PARP2 (BPS Bio #78296).
  • PARP1 assay samples were treated at a concentration of 0.1 nM, 0.3 nM, 1 nM, 10 nM, 100 nM, 1,000 nM, and 10,000 nM.
  • PARP2 assay samples were treated at a concentration of 10 nM, 30 nM, 100 nM, 300 nM, 1,000 nM, 3,000 nM, and 10,000 nM. After sample treatment, the reaction was allowed to proceed for 60 minutes at room temperature, and fluorescence polarization (Ex 485 nm/Em 535 nm) was measured. 2.
  • the PARP1/PARP2 binding ability EC50 (nM) obtained according to the above experimental method is shown in a following table.
  • the compounds according to examples of the present invention exhibit an excellent inhibitory effect against PARP1 and an excellent selective inhibitory effect against PARP1.

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Abstract

The present invention relates to a compound represented by formula 1 as a PARP1 inhibitor, stereoisomers thereof, pharmaceutically acceptable salts thereof and a pharmaceutical composition including the same, and a method for preventing or treating PARP1 activity-associated diseases using the same and a use thereof.

Description

DESCRIPTION
Title
COMPOUND AS PARP1 INHIBITOR AND PHARMACEUTICAL COMPOSITION COMPRISING THE SAME
Technical Field
The present invention relates to a compound, stereoisomers thereof, pharmaceutically acceptable salts thereof as a PARP1 inhibitor and a pharmaceutical composition including the same, and a method for preventing or treating PARP1 activity- associated diseases using the same and a use thereof.
Background
Enzymes of poly(ADP-ribose)polymerase (PARP) family play important roles in many cellular processes such as replication, recombination, chromatin remodeling, and DNA damage repair (O'Connor MJ, Mol Cell (2015) 60(4):547-60).
Examples of PARP inhibitors and their mechanisms of action are instructed, for example, in WO 2004/080976.
PARP1 and PARP2 are the most extensively studied PARPs for their roles in DNA damage repair. PARP1 is activated by DNA damage break and functions to catalyze the addition of poly(ADP-ribose) (PAR) chains to a target protein. This post-translational modification, known as PARylation, mediates recruitment of additional DNA repair factors into DNA lesions.
After completion of this recruitment role, PARP self-PARylation triggers release of bound PARP from DNA, thus allowing access to other DNA repair proteins to complete the repair. Thus, the binding of PARP to an injured site, catalytic activity thereof, and eventual release thereof from DNA are all important steps in which cancer cells respond to DNA damage caused by chemotherapy and radiation therapy (Bai P. Biology of poly(ADP-ribose) polymerases: the factotums of cell maintenance. Mol Cell 2015;58:947-58.).
Inhibition of the PARP family enzyme was utilized as a strategy to selectively kill cancer cells by inactivating complementary DNA repair pathways. A number of preclinical and clinical studies have demonstrated that tumor cells, including deleterious alterations of BRCA1 or BRCA2, the major tumor suppressor proteins involved in double-strand DNA break (DSB) repair by homologous recombination (HR), are selectively sensitive to small molecule inhibitors of DNA repair enzymes of the PARP family. These tumors have a defective homologous recombination repair (HRR) pathway and rely on PARP enzyme function for survival. PARP inhibitor therapy has mainly targeted BRCA-mutant cancer, but PARP inhibitors have been clinically tested in non-BRCA mutant tumors exhibiting homologous recombination deficiency (HRD) (Turner N, Tutt A, Ashworth A. Hallmarks of 'BRCAness' in sporadic cancers. Nat Rev Cancer 2004;4:814-9.).
It is believed that PARP inhibitors with improved selectivity for PARP1 can lead to improved efficacy and reduced toxicity compared to other clinical PARP 1/2 inhibitors. It is also believed that selectively and potently inhibition of PARP1 causes the capture of PARP1 on DNA, thus leading to double-strand DNA break (DSB) through the disruption of S-phase replication fork. PARP 1 -DNA capture is also believed to be an effective mechanism for selectively killing tumor cells with HRD.
Thus, there is an unmet medical need for effective and safe PARP inhibitors, particularly PARP inhibitors with selectivity for PARP1.
[Related Art References]
[Patent Documents]
(Patent Document 0001) International Patent Publication No. WO 2021/013735 (Patent Document 0002) International Patent Publication No. WO 2021/260092 [Non-Patent Documents]
(Non-Patent Document 0001) J. Med. Chem. 2021, 64, 19, 14498-14512
Disclosure
Technical Problem
The present invention may provide a compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof as a PARP1 inhibitor.
The present invention may provide a pharmaceutical composition including a compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof as a PARP1 inhibitor.
The present invention may provide a pharmaceutical composition including a compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof as a PARP1 inhibitor as an active ingredient for preventing or treating PARP1 activity-associated diseases.
The present invention may provide a method for preventing or treating PARP1 activity-associated diseases including administering a compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof as a PARP1 inhibitor.
The present invention may provide a use of a compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof as a PARP1 inhibitor for preventing or treating PARP1 activity-associated diseases.
The present invention may provide a use of a compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof as a PARP1 inhibitor in preparing a medicament for preventing or treating PARP1 activity-associated diseases.
Technical Solution
Hereinafter, the present invention will be described in more detail. All the combinations of various elements disclosed in the present invention fall within the scope of the present invention. In addition, it cannot be seen that the scope of the present invention is limited to the specific description below.
Compound represented by formula 1
The present invention may provide a compound represented by a following formula
1, stereoisomers thereof, or pharmaceutically acceptable salts thereof:
[Formula 1] wherein,
M is CRX, N, NRy, O, or S,
X, Y and Z are each independently N or CRZ, each of Rx, Ry and Rz is independently H, halogen, C1-C5 alkyl, or C1-C5 haloalkyl,
RA and RB are each independently H, halogen, OH, CN, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, or O-(C1-C5 alkyl), or RA and RB may be bonded to each other to form cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl, is a single bond or a double bond, is absent or a single bond,
Ri and R2 are each independently H, halogen, OH, CN, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, or O-(C1-C5 alkyl), A is cycloalkylene, cycloalkenylene, heterocycloalkylene, heterocycloalkenylene, arylene, or heteroarylene, and halogen is F, Cl, Br or I.
In one embodiment, in above formula 1,
M is CRX, N, NRy, O, or S,
X, Y and Z are each independently N or CRZ, each of Rx, Ry and Rz is independently H, halogen, C1-C5 alkyl, or C1-C5 haloalkyl, RA and RB are each independently H, halogen, OH, CN, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, or O-(C1-C5 alkyl), or RA and RB may be bonded to each other to form three- to seven-membered cycloalkyl, three- to seven-membered cycloalkenyl, three- to seven-membered heterocycloalkyl including one to three heteroatoms independently selected from the group consisting of N, O and S, or three- to seven-membered heterocycloalkenyl including one to three heteroatoms independently selected from the group consisting of N, O and S, is a single bond or a double bond, is absent or a single bond,
Ri and R2 are each independently H, halogen, OH, CN, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, or O-(C1-C5 alkyl),
A is three- to seven-membered cycloalkylene, three- to seven-membered cycloalkenylene, three- to seven-membered heterocycloalkylene including one to three heteroatoms independently selected from the group consisting of N, O and S, three- to sevenmembered heterocycloalkenylene including one to three heteroatoms independently selected from the group consisting of N, O and S, C6-C12 arylene, or five- to 12-membered heteroarylene including one to three heteroatoms independently selected from the group consisting of N, O and S, and halogen may be F, Cl, Br or I.
In one embodiment, in above formula 1,
M is CRX, N, O or S,
X, Y and Z are each independently N or CRZ, each of Rx and Rz is independently H or halogen,
RA and RB are each independently H or C1-C5 alkyl, or RA and RB may be bonded to each other to form three- to seven-membered cycloalkyl,
“““ is a single bond or a double bond, is absent or a single bond,
Ri and R2 are each independently H, halogen, C1-C5 alkyl, C1-C5 haloalkyl, or O- (C1-C5 alkyl),
A is C6-C12 arylene or five- to 12-membered heteroarylene including one to three heteroatoms independently selected from the group consisting of N, O and S, and halogen may be F.
In one embodiment, in above formula 1,
M is CRX, N, O or S,
X, Y and Z are each independently N or CRZ, each of Rx and Rz is independently H or halogen,
RA and RB are each independently H or C1-C3 alkyl, or RA and RB may be bonded to each other to form four- to six-membered cycloalkyl, is a single bond or a double bond, is absent or a single bond,
Ri and R2 are each independently H, halogen, C1-C3 alkyl, C1-C3 haloalkyl, or O- (C1-C3 alkyl),
A is phenylene or five- to six-membered heteroarylene including one to three Ns, and halogen may be F.
In the present invention, "Cm-Cn" (in which m and n are each independently an integer of 1 or more) may mean the number of carbons, for example, "C1-C5 alkyl" may represent alkyl having one to five carbon atoms.
In the present invention, "alkyl" may mean a linear or branched saturated hydrocarbon group. In the present invention, alkyl may have one to five carbon atoms. In one embodiment, alkyl may have one to four carbon atoms. In one embodiment, alkyl may have one to three carbon atoms. Examples of alkyl may include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, n-pentyl, sec -pentyl, tert-pentyl, isopentyl, sec-isopentyl, neo-pentyl, and the like, but are not limited thereto.
In the present invention, "alkenyl" may refer to a linear or branched unsaturated hydrocarbon group including one or more double bonds. In the present invention, alkenyl may have two to five carbon atoms. In one embodiment, alkenyl may have two to four carbon atoms. Examples of alkenyl may include ethenyl, allyl, propenyl, and the like, but are not limited thereto.
In the present invention, "alkynyl" may refer to a linear or branched unsaturated hydrocarbon group including one or more triple bonds. In the present invention, alkynyl may have two to five carbon atoms. In one embodiment, alkynyl may have two to four carbon atoms. Examples of alkynyl may include ethynyl, propynyl, butynyl, and the like, but are not limited thereto.
In the present invention, "haloalkyl" may refer to alkyl substituted with one or more halogens such as F, Cl, Br, and I. Alkyl may be the same as defined above.
In the present invention, "cycloalkyl" may mean a saturated hydrocarbon ring having three or more carbon atoms, which may include both monocyclic and polycyclic structures. As one example, in the present invention, cycloalkyl may be a ring having three to 12, three to ten, three to eight, or three to seven carbon atoms. Examples of cycloalkyl may include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclopentyl, bicyclohexyl, bicycloheptyl, bicyclooctyl, bicyclononyl, spiropentyl, spirohexyl, spiroheptyl, spirooctyl, spirononyl, and the like, but are not limited thereto.
In the present invention, "cycloalkenyl" may mean an unsaturated hydrocarbon ring having three or more carbon atoms including one or more double bonds, which may include both monocyclic and polycyclic structures. In other words, in the present invention, cycloalkenyl may mean a ring structure including one or more carbon-carbon double bonds in the above-defined cycloalkyl ring. In the present invention, cycloalkenyl may be a ring having three to 12, three to ten, three to eight, or three to seven carbon atoms. Examples of cycloalkenyl may include cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, and the like, but are not limited thereto.
In the present invention, "heterocyclo alkyl" may mean a cyclic functional group in which at least one or more carbon atoms constituting the above-defined cycloalkyl ring are substituted with a heteroatom. Examples of the heteroatom may include nitrogen (N), oxygen (O), or sulfur (S). In this case, the heteroatoms included in the ring of heterocycloalkyl may be one type or two or more types, one or one or more of one type of heteroatom may be included therein, and at least one or more of the two or more types of heteroatoms may be included therein, respectively. As one example, in the present invention, heterocycloalkyl may be a three- to 12-membered ring. As one example, in the present invention, heterocycloalkyl may be a three- to seven-membered ring including one to three heteroatoms. As one example, in the present invention, heterocycloalkyl may be a four- to six-membered ring having one or two heteroatoms. Examples of heterocycloalkyl may include oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, morpholinyl, piperazinyl, thiomorpholinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like, but are not limited thereto.
In the present invention, "heterocycloalkenyl" may mean a cyclic functional group in which at least one or more carbon atoms constituting the above-defined cycloalkenyl ring are substituted with a heteroatom. Examples of the heteroatom may include nitrogen (N), oxygen (O), or sulfur (S). In this case, the heteroatoms included in the ring of heterocycloalkenyl may be one type or two or more types, one or one or more of one type of heteroatom may be included therein, and at least one or more of the two or more types of heteroatoms may be included therein, respectively.
In the present invention, "aryl" may mean a monocyclic or polycyclic aromatic hydrocarbon ring. As one example, in the present invention, aryl may have six to 20 carbon atoms. As one example, in the present invention, aryl may have six to 12 carbon atoms. Examples of aryl may include phenyl, biphenyl, naphthalenyl, and the like, but are not limited thereto.
In the present invention, “heteroaryl” may mean a monocyclic or polycyclic aromatic hydrocarbon ring including at least one or more heteroatoms such as nitrogen (N), oxygen (O), or sulfur (S). As one example, in the present invention, heteroaryl may be a five- to 12- membered ring. As one example, in the present invention, heteroaryl may be a five- to 12- membered ring having one to three heteroatoms. As one example, in the present invention, heteroaryl may be a five- to eight-membered ring having one to three heteroatoms. As one example, in the present invention, heteroaryl may be a five- or six-membered ring having one or two heteroatoms. As one example, in the present invention, heteroaryl may be a five- or sixmembered ring having three Ns. When heteroaryl includes two or more heteroatoms, the types of heteroatoms may be the same as or different from each other. For example, a case in which heteroaryl includes two or more heteroatoms selected from nitrogen, oxygen, and sulfur may refer to various combinations such as a case of including two nitrogen atoms, a case of including one nitrogen atom and one oxygen atom, a case of including two oxygen atoms and one nitrogen atom, and the like. Examples of heteroaryl may include pyridinyl, thiophenyl, triazolyl, tetrazolyl, benzothiazolyl, benzothiophenyl, quinolinyl, indolyl, isoindolyl, benzofuranyl, benzopyrroyl, furanyl, pyrrolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrazinyl, pyridazinyl, pyrimidinyl, isoquinolinyl, benzoxazolyl, benzoimidazolyl, dihydrobenzothiophenyl, purinyl, indolizinyl, chromenyl, pyrrolopyridinyl, pyrazolopyridinyl, thiadiazolopyridinyl, triazinyl, triazolopyrimidinyl, triazolopyridinyl, triazolopyridazinyl, indazolyl, imidazopyridinyl, imidazopyridazinyl, oxadiazolopyridinyl, benzothiadiazolyl, benzotriazolyl, benzoxadiazole, isomers thereof, and the like, but are not limited thereto. In the present invention, cycloalkylene, cycloalkenylene, heterocycloalkylene, heterocycloalkenylene, arylene, heteroarylene, or phenylene may mean a divalent functional group which is further free of one hydrogen in cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, or phenyl.
The present invention may provide a compound described in a following table, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
In the present invention, “stereoisomer” may include a diastereomer and an optical isomer, which may include a single enantiomer, a mixture of enantiomers including a racemate, a single diastereomer, and a mixture of diastereomers all. Such isomer may be separated by resolution according to conventional techniques, for example, column chromatography, HPLC, or the like. Alternatively, the isomer may be stereospecifically synthesized by using a known array of optically pure starting materials and/or reagents. Specifically, the isomer may be an optical isomer.
In the present invention, “pharmaceutically acceptable salts” may mean salts conventionally used in a pharmaceutical field, and may include, for example, inorganic ion salts prepared from calcium, potassium, sodium, magnesium, and the like; inorganic acid salts prepared from hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, sulfuric acid, hydroiodic acid, etc.; organic acid salts prepared from acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbric acid, carbonic acid, vanillic acid, etc.; sulfonic acid salts prepared from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, and the like; amino acid salts prepared from glycine, arginine, lysine, etc.; amine salts prepared from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc.; and the like, but are not limited thereto.
The compound represented by formula 1 of the present invention may be the same as the compound list in this specification, but also include a pharmaceutically acceptable isotopic-labeled compound in which at least one or more atoms are replaced with an atom having the same atomic number but having an atomic mass or mass number different from the atomic mass or mass number prevailing in nature. Examples of isotopes which may be included in the compound of the present invention may include: isotope of hydrogen, 2H, 3H; isotope of carbon, nC, 13C, 14C; isotope of chlorine, 36Cl;isotope of fluorine, 18F; isotope of iodine, 123I, 125I; isotope of nitrogen, 13N, 15N; isotope of oxygen, 150, 170, 18O; isotope of phosphorus, 32P; isotope of sulfur, 35S; and the like. A certain isotopic-labeled compound of the present invention, for example, a compound with radioactive isotopes incorporated, may be useful in studying drugs and/or a distribution of substrate tissues (e.g., assays). A radioactive isotope tritium, that is, 3H, and carbon- 14, that is, 14C may be useful in view of ease of incorporation and means of immediate detection. Substitution with heavier isotopes, for example, substitution of hydrogen (XH) with deuterium (2H), may exhibit an excellent therapeutic effect on diseases by enhancing metabolic stability, such as increasing a half-life in vivo or reducing a dosage. Substitution with positron-emitting isotopes, for example, nC, 15F, 18F, 15O, 13N, etc., may be useful in studying positron emission tomography (PET) to examine a substrate receptor occupancy. An isotopic-labeled compound of the present invention may be generally prepared by conventional techniques known to those skilled in the art or by processes similar to those described in the schemes and/or examples and preparation examples described in this specification, using an appropriate isotopic-labeled reagent instead of the non-labeled reagent as used in this specification. The compound represented by formula 1 in the present specification may include isotopic-labeled compounds, for example, including, but not limited to, isotopes such as deuterium and tritium, and all other isotopes discussed above.
In the present invention, “prevention” may refer to all the acts, which inhibit or delay the occurrence of a disease by administering the compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
In the present invention, “treatment” may refer to all the acts, by which a symptom of an individual likely to develop or suffering from a disease gets better or takes a favorable turn by administering the compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
The compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof may be advantageously used for preventing or treating PARP1 activity-associated diseases.
The compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof may inhibit PARP1 activity.
The compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof may show an effect of preventing or treating the PARP1 activity-associated diseases at a level similar to, substantially the same as, or higher than that of a conventionally known drug for preventing or treating the PARP1 activity-associated diseases.
Method for preparing compound represented by formula 1
The present invention may provide a method for preparing the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
The compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof may be prepared according to any one method of reaction formulas 1 to 4, which may be modified to a level obvious to those skilled in the art.
In following reaction formulas 1 to 4, Halo may mean F, Cl, Br or I, and each of Ri, R2, RA, RB, A, X, Y, Z and M may be substantially the same as defined in formula 1, if not specifically defined.
[Reaction Formula 1]
According to the reaction formula 1, a compound of formula 1-1 may be subjected to a substitution reaction with an amine compound to prepare a compound of formula 1-2, which may be subjected to a Suzuki reaction to prepare a compound of formula 1-3, and a protecting group may be removed to prepare a compound of formula 1-4.
[Reaction Formula 2]
According to the reaction formula 2, the compound of formula 1-1 may be subjected to a Suzuki reaction to prepare a compound of formula 2-1, which may be subjected to a substitution reaction with the amine compound to prepare the compound of formula 1-3, and a protecting group may be removed to prepare the compound of formula 1-4.
[Reaction Formula 3]
According to the reaction formula 3, the compound of formula 1-1 may be subjected to a Suzuki reaction to prepare the compound of formula 2-1, which may be subjected to a hydrolysis reaction to prepare a compound of formula 3-1, which may be subjected to a substitution reaction with the amine compound to prepare the compound of formula 1-3, and a protecting group may be removed to prepare the compound of formula 1-4.
[Reaction Formula 4]
According to the reaction formula 4, a compound of formula 4-1 and the compound of formula 1-4 may be subjected to a substitution reaction, or a compound of formula 4-2 and the compound of formula 1-4 may be subjected to a reductive amination reaction to prepare a compound of formula 4-3.
In the present invention, the compounds prepared by the reaction formula 4 may include example compounds 1 to 88.
Pharmaceutical composition, method and use
The present invention may provide a pharmaceutical composition including the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
The present invention may provide a pharmaceutical composition including the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient for preventing or treating PARP1 activity-associated diseases.
In other words, the pharmaceutical composition including the compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient may be advantageously used for preventing or treating
PARP1 activity-associated diseases. The PARP1 activity-associated diseases may be cancer or inflammatory diseases.
The cancer may be at least one selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, hematological cancer, gastrointestinal cancer such as gastric cancer or colorectal cancer, lung cancer such as small cell lung cancer or non-small cell lung cancer, and brain cancer such as glioma or glioblastoma.
The brain cancer may be metastatic cancer caused by tumors in other parts of the body, such as breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, hematological cancer, gastrointestinal cancer such as gastric cancer and colorectal cancer, or lung cancer such as small cell lung cancer or non-small cell lung cancer.
The inflammatory disease may be at least one selected from the group consisting of rheumatoid arthritis, multiple sclerosis, Crohn's disease, ulcerative colonitis, ulcerative colitis, graft-versus-host disease (GVHD), systemic lupus erythematosus, toxic shock syndrome, osteoarthritis, and insulin-dependent diabetes mellitus.
For administration, the pharmaceutical composition of the present invention may further include at least one pharmaceutically acceptable carrier, in addition to the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof. The pharmaceutically acceptable carrier may be one which is conventionally used in the art, specifically including, but not limited thereto, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidine, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, or mineral oil. The pharmaceutical composition of the present invention may further include lubricant, humectant, sweetening agent, flavoring agent, emulsifier, suspending agent, preservative, dispersing agent, stabilizing agent, etc., in addition to the above ingredients. In addition, the pharmaceutical composition of the present invention may be formulated into an oral dosage form such as tablet, powder, granule, pill, capsule, suspension, emulsion, liquid for internal use, emulsion, syrup, etc., as well as a form of external preparation, suppository and sterile solution for injection by using a pharmaceutically acceptable carrier and excipient, and thus may be prepared in a unit dose form or prepared by being inserted into a multi-dose container. Preparations may be formulated according to a conventional method used for formulation in the art or a method disclosed in Remington's Pharmaceutical Science (19th ed., 1995), and may be formulated into various preparations depending on each disease or ingredient.
The pharmaceutical composition of the present invention may be orally or parenterally administered (for example, applied intravenously, hypodermically, intraperitoneally or locally) according to a targeted method, in which a dosage thereof may vary in a range thereof depending on a patient’s weight, age, gender, health condition and diet, an administration time, an administration method, an excretion rate, a severity of a disease and the like. The compound represented by formula 1 of the present invention may be administered once or several divided times a day, but is not necessarily limited thereto.
A daily dosage of the compound represented by formula 1 of the present invention, stereoisomers thereof, or pharmaceutically acceptable salts thereof may be specifically about 0.1 to about 10,000 mg/kg, about 1 to about 8,000 mg/kg, about 5 to about 6,000 mg/kg, or about 10 to about 4,000 mg/kg, and more specifically about 50 to about 2,000 mg/kg, but is not limited thereto.
In addition to the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof, the pharmaceutical composition of the present invention may further include at least one ingredient which may exhibit the same or similar medicinal effects or may bring synergy to medicinal effects in combination. The pharmaceutical composition of the present invention may be administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent.
The present invention may provide a method for preventing or treating PARP1 activity-associated diseases including administering the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
In the present invention, “administration” may refer to introducing a predetermined substance into an individual by an appropriate method.
In the present invention, “individual” may refer to all the animals such as rats, mice, livestock, etc., including humans, who are likely to develop or have already developed diseases, and specifically refer to mammals including humans, but is not limited thereto.
The method for preventing or treating PARP1 activity-associated diseases of the present invention may include administering a therapeutically effective amount of the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof.
In the present invention, “therapeutically effective amount” may refer to an amount enough to treat a disease at a reasonable risk/benefit ratio applicable to medical treatment and not to cause a side effect, and may be determined by those skilled in the art according to factors including a patient’s gender, age, weight and health condition, a type of disease, severity, activity of a drug, sensitivity to a drug, an administration method, an administration time, an administration route, an excretion rate, a treatment period, a drug combined or concurrently used, as well as other factors well known in a pharmaceutical field. It may be preferable that a specific therapeutically effective amount is to be differently applied to each certain patient depending on various factors including a type and degree of reaction to be achieved therefrom, a specific composition including whether other preparations are used in some cases, a patient’s age, weight, general health condition, gender and diet, an administration time, an administration route, a secretion rate of the composition, a treatment period and a drug used together with the specific composition or simultaneously therewith, as well as other similar factors well known in a pharmaceutical field.
The present invention may provide a use of the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof for preventing or treating PARP1 activity-associated diseases.
The present invention may provide a use of the compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof in preparing a medicament for preventing or treating PARP1 activity-associated diseases.
The present invention may provide a use of the pharmaceutical composition according to the present invention for preventing or treating PARP1 activity-associated diseases.
The present invention may provide a use of the pharmaceutical composition according to the present invention in preparing a medicament for preventing or treating PARP1 activity-associated diseases.
Matters mentioned in the compound represented by formula 1, pharmaceutical composition, method and use of the present invention may be applied the same, if not contradictory to each other.
Advantageous Effects
The compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof of the present invention may show an inhibitory effect against PARP1 and may be advantageously used for preventing or treating PARP1 activity-associated diseases. Mode for Invention
Hereinafter, the present invention will be described in more detail through preparation examples and exemplary examples. However, the following preparation examples and exemplary examples are provided for the purpose of illustrating the present invention, and thus the present invention is not limited to the preparation examples and exemplary examples.
Preparation Example>
The compounds of the present invention were synthesized as follows.
In order to prepare compounds of the present invention, each of the reacting materials used in each reaction was purchased from Sigma Aldrich, etc., or was synthesized by using an organic synthesis method obvious to those skilled in the chemistry field, and was used without a separate purification process. The compounds of each example were identified through 1 H- NMR (Bruker, avance II 400) and Mass (Waters, SQD2) analysis.
Preparation of intermediate compound
Synthesis of intermediate compound 1: 5-(3,6-dihydro-2H-oxazin-5-yl)-N- methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of 5-bromo-N-methyl-pyridin-2-carboxamide
Methyl 5-bromopyridin-2-carboxylate (100.00%, 1.000 g, 4.629 mmol) and methylamine solution (2.0 M solution in methanol, 10 mL, 20.000 mmol) were mixed at room temperature, and the resulting reaction mixture was stirred at the same temperature for two hours. Solvent was removed from the reaction mixture under reduced pressure, and then a precipitated solid was filtered out, washed with hexane, and dried to obtain 5-bromo-N-methyl- pyridin-2-carboxamide (923.400 mg, 92.76%) in a white solid form.
[Step 2] Synthesis of tert-butyl 5-[6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which 5-bromo-N-methyl-pyridin-2-carboxamide (100.00%, 200.000 mg, 0.930 mmol), tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 304.000 mg, 0.977 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 61.000 mg, 0.094 mmol), and potassium carbonate (100.00%, 257.000 mg, 1.860 mmol) were dissolved in 1,4-dioxane (9 mL)/water (3 mL) was stirred at room temperature for 18 hours and further stirred at 100°C for one hour, after which the temperature was lowered to room temperature to terminate the reaction. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; ethyl acetate/hexane = 0 to 50%) and concentrated to obtain tert-butyl 5-[6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (258.000 mg, 86.85%) in a white solid form.
[Step 3] Synthesis of intermediate compound 1
To a solution in which tert-butyl 5-[6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 258.000 mg, 0.808 mmol) was dissolved in dichloromethane (10 mL) at room temperature, trifluoroacetic acid (100.00% solution, 1.85 mL, 24.176 mmol) was added and stirred at the same temperature for 30 minutes. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 40 g cartridge; methanol/dichloromethane = 5 to 20%) and concentrated to obtain intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (256.900 mg, 95.42%) in a white solid form.
Synthesis of intermediate compound 2: 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of methyl 5-bromo-6-fluoro-pyridin-2-carboxylate
A solution in which methyl 5-bromopyridin-2-carboxylate (100.00%, 3.000 g, 13.887 mmol) and silver(II) fluoride (100.00%, 15.190 g, 104.137 mmol) were dissolved in acetonitrile (20 mL) at room temperature was stirred at the same temperature. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCL, 40 g cartridge; ethyl acetate/hexane = 0 to 20%) and concentrated, and then the resulting product was crystallized with ethyl acetate (10 mL) and hexane (20 mL) at 25 °C and filtered to obtain a solid, which was then washed with hexane and dried to obtain methyl 5-bromo-6-fluoro-pyridin-2-carboxylate (1.660 g, 51.08%) in a white solid form.
[Step 2] Synthesis of tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 1.300 g, 4.177 mmol), methyl 5-bromo-6-fluoro- pyridin-2-carboxylate (100.00%, 1.000 g, 4.273 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 272.000 mg, 0.417 mmol), and potassium carbonate (100.00%, 1.150 g, 8.321 mmol) were dissolved in 1,4-dioxane (18 mL)/water (6 mL) at room temperature was stirred at 100°C for one hour, after which the temperature was lowered to room temperature to terminate the reaction. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 40 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (775.000 mg, 54.84%) in an ivory solid form.
[Step 3] Synthesis of tert-butyl 5-[2-fluoro-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 100.000 mg, 0.296 mmol) and methylamine solution (2.0 M solution in THF, 0.3 mL, 0.600 mmol) were dissolved in methanol (1 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-[2-fluoro-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (92.800 mg, 93.05%) in a white solid form.
[Step 4] Synthesis of intermediate compound 2
To a solution in which tert-butyl 5-[2-fluoro-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 62.000 mg, 0.184 mmol) was dissolved in methanol (1 mL) at room temperature, trifluoroacetic acid (100.00% solution, 1.2 mL, 9.693 mmol) was added and stirred overnight at 50°C, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 0 to 10%) and concentrated to obtain intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6-fluoro-N-methyl-pyridin-2- carboxamide; 2,2,2-trifluoroacetic acid (48.300 mg, 74.84%) in a white solid form. Synthesis of intermediate compound 3: 4-(3,6-dihydro-2H-oxazin-5-yl)-N- methyl-benzamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-[4-(methylcarbamoyl)phenyl]-3,6-dihydrooxazin-
2-carboxylate
A solution in which 4-bromo-N-methyl-benzamide (100.00%, 200.000 mg, 0.934 mmol), tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydrooxazin-2- carboxylate (100.00%, 305.000 mg, 0.980 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 61.000 mg, 0.094 mmol), and potassium carbonate (100.00%, 258.000 mg, 1.867 mmol) were dissolved in 1,4-dioxane (2 mL)/water (0.5 m ) at room temperature was stirred at 100°C for one hour, after which the temperature was lowered to room temperature to terminate the reaction. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; ethyl acetate/hexane = 0 to 70%) and concentrated to obtain tert-butyl 5-[4-(methylcarbamoyl)phenyl]-3,6-dihydrooxazin-2- carboxylate (254.000 mg, 85.38%) in a brown solid form.
[Step 2] Synthesis of intermediate compound 3
A solution in which tert-butyl 5-[4-(methylcarbamoyl)phenyl]-3,6-dihydrooxazin-2- carboxylate (100.00%, 254.000 mg, 0.798 mmol) and trifluoro acetic acid (100.00% solution, 2.5 mL, 32.670 mmol) were dissolved in dichloromethane (10 mL) at room temperature was stirred at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then a precipitated solid was filtered out, washed with dichloromethane, and dried to obtain intermediate compound 3 of 4-(3,6-dihydro-2H-oxazin-5-yl)-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (351.000 mg, 98.59%) in a white solid form.
Synthesis of intermediate compound 4: 5-(3,6-dihydro-2H-oxazin-5-yl)-N- methyl-pyrimidin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-[2-(methylcarbamoyl)pyrimidin-5-yl]-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 500.000 mg, 1.607 mmol), 5-bromo-N-methyl- pyrimidin-2-carboxamide (100.00%, 366.000 mg, 1.694 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 105.000 mg, 0.161 mmol), and potassium carbonate (100.00%, 444.000 mg, 3.213 mmol) were dissolved in 1,4-dioxane (12 mL)/water (3 mL) at room temperature was stirred at the same temperature. Water was poured into the reaction mixture and extracted with dichloromethane. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated to obtain tert-butyl 5-[2-(methylcarbamoyl)pyrimidin-5-yl]-3,6- dihydrooxazin-2-carboxylate (139.000 mg, 27.01%) in a brown solid form.
[Step 2] Synthesis of intermediate compound 4
A solution in which tert-butyl 5-[2-(methylcarbamoyl)pyrimidin-5-yl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 460.000 mg, 1.436 mmol) and trifluoroacetic acid (100.00% solution, 5.3 mL, 42.811 mmol) were dissolved in tetrahydrofuran (5 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; methanol/dichloromethane = 0 to 30%) and concentrated to obtain intermediate compound 4 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N-methyl- pyrimidin-2-carboxamide; 2,2,2-trifluoroacetic acid (320.000 mg, 66.66%) in a brown oil form.
Synthesis of intermediate compound 5: 5-(3,6-dihydro-2H-oxazin-5-yl)-N,3- dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-(6-ethoxycarbonyl-5-methyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.500 g, 2.000 mmol), ethyl 5-bromo-3-methyl- pyridin-2-carboxylate (100.00%, 0.412 g, 1.687 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 0.105 g, 0.161 mmol), and potassium carbonate (100.00%, 0.444 g, 3.213 mmol) were dissolved in 1,4-dioxane (12 mL)/water (3 mL) at 100°C was stirred at the same temperature for two hours, after which the temperature was lowered to room temperature to terminate the reaction. The reaction mixture was filtered via a celite pad to remove a solid therefrom, and then solvent was removed from the resulting filtrate without the solid under reduced pressure. Then, the resulting concentrate was purified via column chromatography (SiCT, 24 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-(6-ethoxycarbonyl-5-methyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (0.400 g, 70.00%) in a white solid form.
[Step 2] Synthesis of tert-butyl 5-[5-methyl-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A reaction mixture in which tert-butyl 5-(6-ethoxycarbonyl-5-methyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.400 g, 1.148 mmol) and methylamine solution (2.0 M solution in THF, 5.741 mL, 11.482 mmol) were mixed at room temperature was stirred at the same temperature for three days. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5- [5-methyl-6-(methylcarbamoyl)-3-pyridyl]-3,6-dihydrooxazin-2-carboxylate (0.250 g, 65.31%) in a white solid form.
[Step 3] Synthesis of intermediate compound 5
A solution in which tert-butyl 5-[5-methyl-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.250 g, 0.750 mmol) and trifluoroacetic acid (100.00% solution, 0.574 mL, 7.496 mmol) were dissolved in dichloromethane (3 mL) at room temperature was stirred at the same temperature for three hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; methanol/dichloromethane = 0 to 20%) and concentrated to obtain intermediate compound 5 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N,3- dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (210.000 mg, 84.53%) in a white solid form.
Synthesis of intermediate compound 6: 5-(3,6-dihydro-2H-oxazin-5-yl)-N- methyl-3-(trifluoromethyl)pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-[6-methoxycarbonyl-5-(trifluoromethyl)-3- pyridyl] -3 ,6-dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 500.000 mg, 1.607 mmol), methyl 5-bromo-3- (trifluoromethyl)pyridin-2-carboxylate (100.00%, 366.000 mg, 1.289 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 105.000 mg, 0.161 mmol), and potassium carbonate (100.00%, 444.000 mg, 3.213 mmol) were dissolved in 1,4-dioxane (12 mL)/water (3 mL) at 100°C was stirred at the same temperature for two hours, after which the temperature was lowered to room temperature to terminate the reaction. The reaction mixture was filtered via a celite pad to remove a solid therefrom, and then solvent was removed from the resulting filtrate without the solid under reduced pressure. Then, the resulting concentrate was purified via column chromatography (SiCL, 24 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-[6-methoxycarbonyl-5-(trifluoromethyl)-3- pyridyl]-3,6-dihydrooxazin-2-carboxylate (0.480 g, 76.94%) in a white solid form.
[Step 2] Synthesis of tert-butyl 5-[6-(methylcarbamoyl)-5-(trifluoromethyl)-3- pyridyl] -3 ,6-dihydrooxazin-2-carboxylate
A reaction mixture in which tert-butyl 5-[6-methoxycarbonyl-5-(trifluoromethyl)-3- pyridyl]-3,6-dihydrooxazin-2-carboxylate (100.00%, 0.480 g, 1.236 mmol) and methylamine solution (2.0 M solution in THF, 6.181 mL, 12.362 mmol) were mixed at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; ethyl acetate/hexane = 0 to 100%) and concentrated to obtain tert-butyl 5-[6-(methylcarbamoyl)-5-(trifluoromethyl)-3-pyridyl]-3,6-dihydrooxazin-2- carboxylate (0.117 g, 24.43%) in a white solid form.
[Step 3] Synthesis of intermediate compound 6
Boc
A solution in which tert-butyl 5-[6-(methylcarbamoyl)-5-(trifluoromethyl)-3- pyridyl] -3, 6-dihydrooxazin-2-carboxylate (100.00%, 0.117 g, 0.302 mmol) and trifluoroacetic acid (100.00% solution, 0.231 mL, 3.017 mmol) were dissolved in dichloromethane (10 mL) at room temperature was stirred at the same temperature for two hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; methanol/dichloromethane = 0 to
20%) and concentrated to obtain intermediate compound 6 of 5-(3,6-dihydro-2H-oxazin-5-yl)-
N-methyl-3-(trifluoromethyl)pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (90.000 mg,
77.34%) in a white solid form.
Synthesis of intermediate compound 7: 5-(3,6-dihydro-2H-oxazin-5-yl)-N- methyl-pyrazin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-(5-methoxycarbonylpyrazin-2-yl)-3,6- dihydrooxazin-2-carboxylate
A solution in which methyl 5-bromopyrazin-2-carboxylate (100.00%, 0.350 g, 1.600 mmol), tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydrooxazin-2- carboxylate (100.00%, 0.527 g, 1.693 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 0.105 g, 0.161 mmol), and potassium carbonate (100.00%, 0.446 g, 3.226 mmol) were dissolved in 1,4-dioxane (12 mL)/water (3 mL) at 100°C was stirred at the same temperature for two hours, after which the temperature was lowered to room temperature to terminate the reaction. The reaction mixture was filtered via a celite pad to remove a solid therefrom, and then solvent was removed from the resulting filtrate without the solid under reduced pressure. Then, the resulting concentrate was purified via column chromatography (SiCL, 24 g cartridge; ethyl acetate/hexane = 0 to 60%) and concentrated to obtain tert-butyl 5-(5-methoxycarbonylpyrazin-2-yl)-3,6- dihydrooxazin-2-carboxylate (0.250 g, 48.00%) in a white solid form.
[Step 2] Synthesis of tert-butyl 5-[5-(methylcarbamoyl)pyrazin-2-yl]-3,6- dihydrooxazin-2-carboxylate
A reaction mixture in which tert-butyl 5-(5-methoxycarbonylpyrazin-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.250 g, 0.778 mmol) and methylamine solution (2.0 M solution in THF, 1.945 mL, 3.890 mmol) were mixed at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; ethyl acetate/hexane = 0 to 100%) and concentrated to obtain tert-butyl 5-[5-(methylcarbamoyl)pyrazin-2-yl]-3,6-dihydrooxazin-2-carboxylate (0.110 g, 44.14%) in a white solid form.
[Step 3] Synthesis of intermediate compound 7
A solution in which tert-butyl 5-[5-(methylcarbamoyl)pyrazin-2-yl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.110 g, 0.343 mmol) and trifluoroacetic acid (100.00% solution, 0.526 mL, 6.869 mmol) were dissolved in dichloromethane (12 mL) at room temperature was stirred at the same temperature for three hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; methanol/dichloromethane = 0 to 20%) and concentrated to obtain intermediate compound 7 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N-methyl- pyrazin-2-carboxamide; 2,2,2-trifluoroacetic acid (0.090 g, 82.35%) in a white solid form.
Synthesis of intermediate compound 8: 6-(3,6-dihydro-2H-oxazin-5-yl)-N- methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-(5-methoxycarbonyl-2-pyridyl)-3,6- dihydrooxazin-2-carboxylate A solution in which methyl 6-bromopyridin-3-carboxylate (100.00%, 500.000 mg, 2.315 mmol), tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydrooxazin-2- carboxylate (100.00%, 756.000 mg, 2.429 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 151.000 mg, 0.232 mmol), and potassium carbonate (100.00%, 640.000 mg, 4.631 mmol) were dissolved in 1,4-dioxane (8 mL)/water (2 mL) at room temperature was stirred at the same temperature. Water was poured into the reaction mixture and extracted with dichloromethane. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCL, 24 g cartridge; ethyl acetate/hexane = 0 to 10%) and concentrated to obtain tert-butyl 5-(5-methoxycarbonyl-2-pyridyl)-3,6-dihydrooxazin-2- carboxylate (625.000 mg, 84.31%) in a brown solid form.
[Step 2] Synthesis of tert-butyl 5-[5-(methylcarbamoyl)-2-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(5-methoxycarbonyl-2-pyridyl)-3,6-dihydrooxazin- 2-carboxylate (100.00%, 620.000 mg, 1.936 mmol) and methylamine solution (2.0 M solution in THF, 4.8 mL, 9.600 mmol) were dissolved in methanol (5 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 24 g cartridge; ethyl acetate/hexane = 0 to 50%) and concentrated to obtain tert-butyl 5-[5-(methylcarbamoyl)-2-pyridyl]-3,6-dihydrooxazin-2-carboxylate (140.000 mg, 22.64%) in a light yellow solid form. [Step 3] Synthesis of intermediate compound 8
To a solution in which tert-butyl 5-[5-(methylcarbamoyl)-2-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 140.000 mg, 0.438 mmol) was dissolved in dichloromethane (3 mL) at room temperature, trifluoroacetic acid (100.00% solution, 1 mL, 13.068 mmol) was added and stirred at the same temperature for 30 minutes. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 5 to 30%) and concentrated to obtain intermediate compound 8 of 6-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid (94.000 mg, 64.34%) in a light green solid form.
Synthesis of intermediate compound 9: 5-(3,6-dihydro-2H-oxazin-5-yl)-N,4- dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-(6-methoxycarbonyl-4-methyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate
A solution in which methyl 5-bromo-4-methyl-pyridin-2-carboxylate (100.00%, 0.200 g, 0.900 mmol), tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.284 g, 0.913 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 0.057 g, 0.087 mmol), and potassium carbonate (100.00%, 0.240 g, 1.739 mmol) were dissolved in 1,4-dioxane (12 mL)/water (3 mL) at 100°C was stirred at the same temperature for two hours, after which the temperature was lowered to room temperature to terminate the reaction. The reaction mixture was filtered via a celite pad to remove a solid therefrom, and then solvent was removed from the resulting filtrate without the solid under reduced pressure. Then, the resulting concentrate was purified via column chromatography (SiCT, 24 g cartridge; ethyl acetate/hexane = 0 to 100%) and concentrated to obtain tert-butyl 5-(6-methoxycarbonyl-4-methyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (0.150 g, 50.00%) in a white solid form.
[Step 2] Synthesis of 5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-4-methyl- pyridin-2-carboxylic acid
A solution in which tert-butyl 5-(6-methoxycarbonyl-4-methyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.238 g, 0.712 mmol) and lithium hydroxide (100.00% solution, 0.0397 mL, 1.430 mmol) were dissolved in tetrahydrofuran (30 mL)/water (10 mL) at room temperature was stirred at the same temperature for two hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 24 g cartridge; methanol/dichloromethane = 0 to 20%) and concentrated to obtain 5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-4-methyl- pyridin-2-carboxylic acid (0.200 g, 87.73%) in a white solid form.
[Step 3] Synthesis of tert-butyl 5-[4-methyl-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which 5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-4-methyl- pyridin-2-carboxylic acid (100.00%, 0.228 g, 0.712 mmol), methylamine solution (2.0 M solution in THF, 1.780 mL, 3.560 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.7459 mL, 4.271 mmol) were dissolved in N,N-dimethylformamide (5 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-[4-methyl-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (0.143 g, 60.30%) in a white solid form.
[Step 4] Synthesis of intermediate compound 9
A solution in which tert-butyl 5-[4-methyl-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.143 g, 0.429 mmol) and trifluoroacetic acid (100.00% solution, 0.985 mL, 12.863 mmol) were dissolved in dichloromethane (12 mL) at room temperature was stirred at the same temperature for three hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; methanol/dichloromethane = 0 to 20%) and concentrated to obtain intermediate compound 9 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N,4- dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (0.100 g, 70.37%) in a white solid form.
Synthesis of intermediate compound 10: 6-(3,6-dihydro-2H-oxazin-5-yl)-5- fluoro-N-methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-(3-fluoro-5-methoxycarbonyl-2-pyridyl)-3,6- dihydrooxazin-2-carboxylate
A solution in which methyl 6-bromo-5-fluoro-pyridin-3-carboxylate (100.00%, 500.000 mg, 2.137 mmol), tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 698.000 mg, 2.243 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 139.000 mg, 0.213 mmol), and potassium carbonate (100.00%, 591.000 mg, 4.276 mmol) were dissolved in 1,4-dioxane (8 mL)/water (2 mb) at room temperature was stirred at the same temperature. Water was poured into the reaction mixture and extracted with dichloromethane. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-(3-fluoro-5-methoxycarbonyl-2-pyridyl)-3,6- dihydrooxazin-2-carboxylate (606.000 mg, 83.84%) in a brown solid form.
[Step 2] Synthesis of tert-butyl 5-[3-fluoro-5-(methylcarbamoyl)-2-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(3-fluoro-5-methoxycarbonyl-2-pyridyl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 600.000 mg, 1.774 mmol) and methylamine solution (2.0 M solution in THF, 4.5 mL, 9.000 mmol) were dissolved in methanol (5 mL) at room temperature was stirred overnight at 50°C, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; ethyl acetate/hexane = 0 to 50%) and concentrated to obtain tert-butyl 5- [3-fluoro-5-(methylcarbamoyl)-2-pyridyl]-3,6-dihydrooxazin-2-carboxylate (146.000 mg, 24.40%) in a white solid form.
[Step 3] Synthesis of intermediate compound 10
To a solution in which tert-butyl 5-[3-fluoro-5-(methylcarbamoyl)-2-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 284.000 mg, 0.842 mmol) was dissolved in dichloromethane (5 mL) at room temperature, trifluoroacetic acid (100.00% solution, 1.93 mL, 25.221 mmol) was added and stirred at the same temperature for 30 minutes. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 40 g cartridge; methanol/dichloromethane = 5 to 30%) and concentrated to obtain intermediate compound 10 of 6-(3,6-dihydro-2H-oxazin-
5-yl)-5-fluoro-N-methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid (199.000 mg, 67.32%) in a white solid form.
Synthesis of intermediate compound 11: 5-(3,6-dihydro-2H-oxazin-5-yl)-N- ethyl-6-fluoro-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of methyl 5-bromo-6-fluoro-pyridin-2-carboxylate
A solution in which methyl 5-bromopyridin-2-carboxylate (100.00%, 3.000 g, 13.887 mmol) and silver(II) fluoride (100.00%, 15.190 g, 104.137 mmol) were dissolved in acetonitrile (20 mL) at room temperature was stirred at the same temperature. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCL, 40 g cartridge; ethyl acetate/hexane = 0 to 20%) and concentrated, and then the resulting product was crystallized with ethyl acetate (10 mL) and hexane (20 mL) at 25 °C and filtered to obtain a solid, which was then washed with hexane and dried to obtain methyl 5-bromo-6-fluoro-pyridin-2-carboxylate (1.660 g, 51.08%) in a white solid form.
[Step 2] Synthesis of tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 1.300 g, 4.177 mmol), methyl 5-bromo-6-fluoro- pyridin-2-carboxylate (100.00%, 1.000 g, 4.273 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 272.000 mg, 0.417 mmol), and potassium carbonate (100.00%, 1.150 g, 8.321 mmol) were dissolved in 1,4-dioxane (18 mL)/water (6 mL) at room temperature was stirred at 100°C for one hour, after which the temperature was lowered to room temperature to terminate the reaction. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 40 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (775.000 mg, 54.84%) in an ivory solid form.
[Step 3] Synthesis of (5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-6-fluoro- pyridin-2-carboxylic acid
A solution in which tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 1.200 g, 3.547 mmol) and lithium hydroxide monohydrate (100.00% solution, 0.4927 mL, 17.730 mmol) were dissolved in tetrahydrofuran (10 mL)/water (10 mL)/methanol (10 mL) at room temperature was stirred at the same temperature. 1 N-hydrochloric acid aqueous solution was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain (5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-6-fluoro- pyridin-2-carboxylic acid (1.000 g, 86.93%) as a white solid form. The resulting product was used without an additional purification process.
[Step 4] Synthesis of tert-butyl 5-[6-(ethylcarbamoyl)-2-fluoro-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which 5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-6-fluoro- pyridin-2-carboxylic acid (100.00%, 400.000 mg, 1.233 mmol), O-(7-azabenzotriazol-l-yl)- N,N,N',N'-tetramethyluronium hexafluorophosphate (100.00%, 703.000 mg, 1.849 mmol), N- ethyldiisopropylamine (100.00% solution, 0.32 mL, 1.837 mmol), and ethylamine solution (2.0 M solution in methanol, 0.65 mL, 1.300 mmol) were dissolved in tetrahydrofuran (10 mL) at room temperature was stirred at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 24 g cartridge; methanol/dichloromethane = 10 to 30%) and concentrated to obtain tert-butyl 5-[6-(ethylcarbamoyl)-2-fluoro-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (407.000 mg, 93.90%) in a white solid form.
[Step 5] Synthesis of intermediate compound 11
A solution in which tert-butyl 5-[6-(ethylcarbamoyl)-2-fluoro-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 400.000 mg, 1.138 mmol) and trifluoroacetic acid
(100.00% solution, 15 mL, 121.160 mmol) were dissolved in tetrahydrofuran (5 mL) at room temperature was stirred at 50°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; methanol/dichloromethane = 0 to 10%) and concentrated to obtain intermediate compound 11 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N-ethyl-6-fluoro-pyridin-2- carboxamide; 2,2,2-trifluoroacetic acid (194.000 mg, 46.65%) in an orange-colored oil form.
Synthesis of intermediate compound 12: 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-propyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of methyl 5-bromo-6-fluoro-pyridin-2-carboxylate
A solution in which methyl 5-bromopyridin-2-carboxylate (100.00%, 3.000 g, 13.887 mmol) and silver(II) fluoride (100.00%, 15.190 g, 104.137 mmol) were dissolved in acetonitrile (20 mL) at room temperature was stirred at the same temperature. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCL, 40 g cartridge; ethyl acetate/hexane = 0 to 20%) and concentrated, and then the resulting product was crystallized with ethyl acetate (10 mL) and hexane (20 mL) at 25 °C and filtered to obtain a solid, which was then washed with hexane and dried to obtain methyl 5-bromo-6-fluoro-pyridin-2-carboxylate (1.660 g, 51.08%) in a white solid form.
[Step 2] Synthesis of tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 1.300 g, 4.177 mmol), methyl 5-bromo-6-fluoro- pyridin-2-carboxylate (100.00%, 1.000 g, 4.273 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 272.000 mg, 0.417 mmol), and potassium carbonate (100.00%, 1.150 g, 8.321 mmol) were dissolved in 1,4-dioxane (18 mL)/water (6 mL) at room temperature was stirred at 100°C for one hour, after which the temperature was lowered to room temperature to terminate the reaction. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 40 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (775.000 mg, 54.84%) in an ivory solid form.
[Step 3] Synthesis of (5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-6-fluoro- pyridin-2-carboxylic acid
A solution in which tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 1.200 g, 3.547 mmol) and lithium hydroxide monohydrate (100.00% solution, 0.4927 mL, 17.730 mmol) were dissolved in tetrahydrofuran (10 mL)/water (10 mL)/methanol (10 mL) at room temperature was stirred at the same temperature. 1 N-hydrochloric acid aqueous solution was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain (5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-6-fluoro- pyridin-2-carboxylic acid (1.000 g, 86.93%) as a white solid form. The resulting product was used without an additional purification process.
[Step 4] Synthesis of tert-butyl 5-[2-fluoro-6-(propylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which 5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-6-fluoro- pyridin-2-carboxylic acid (100.00%, 0.200 g, 0.600 mmol), O-(7-azabenzotriazol-l-yl)- N,N,N',N'-tetramethyluronium hexafluorophosphate (100.00%, 0.352 g, 0.925 mmol), N- ethyldiisopropylamine (100.00% solution, 0.1612 mL, 0.925 mmol), and propan- 1 -amine (100.00% solution, 0.05330 mL, 0.648 mmol) were dissolved in N,N-dimethylformamide (10 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 24 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated to obtain tert-butyl 5-[2-fluoro-6-(propylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (0.187 g, 80.00%) in a white solid form.
[Step 5] Synthesis of intermediate compound 12
A solution in which tert-butyl 5-[2-fluoro-6-(propylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.187 g, 0.512 mmol) and trifluoroacetic acid (100.00% solution, 0.3919 mL, 5.118 mmol) were dissolved in dichloromethane (10 mL) at room temperature was stirred at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 24 g cartridge; methanol/dichloromethane = 10 to 30%) and concentrated to obtain intermediate compound 12 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6-fluoro- N-propyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (0.100 g, 53.80%) in a white solid form.
Synthesis of intermediate compound 13: 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-isopropyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of methyl 5-bromo-6-fluoro-pyridin-2-carboxylate
A solution in which methyl 5-bromopyridin-2-carboxylate (100.00%, 3.000 g, 13.887 mmol) and silver(II) fluoride (100.00%, 15.190 g, 104.137 mmol) were dissolved in acetonitrile (20 mL) at room temperature was stirred at the same temperature. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCL, 40 g cartridge; ethyl acetate/hexane = 0 to 20%) and concentrated, and then the resulting product was crystallized with ethyl acetate (10 mL) and hexane (20 mL) at 25 °C and filtered to obtain a solid, which was then washed with hexane and dried to obtain methyl 5-bromo-6-fluoro-pyridin-2-carboxylate (1.660 g, 51.08%) in a white solid form.
[Step 2] Synthesis of tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 1.300 g, 4.177 mmol), methyl 5-bromo-6-fluoro- pyridin-2-carboxylate (100.00%, 1.000 g, 4.273 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 272.000 mg, 0.417 mmol), and potassium carbonate (100.00%, 1.150 g, 8.321 mmol) were dissolved in 1,4-dioxane (18 mL)/water (6 mL) at room temperature was stirred at 100°C for one hour, after which the temperature was lowered to room temperature to terminate the reaction. Water was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 40 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (775.000 mg, 54.84%) in an ivory solid form.
[Step 3] Synthesis of (5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-6-fluoro- pyridin-2-carboxylic acid
A solution in which tert-butyl 5-(2-fluoro-6-methoxycarbonyl-3-pyridyl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 1.200 g, 3.547 mmol) and lithium hydroxide monohydrate (100.00% solution, 0.4927 mL, 17.730 mmol) were dissolved in tetrahydrofuran (10 mL)/water (10 mL)/methanol (10 mL) at room temperature was stirred at the same temperature. 1 N-hydrochloric acid aqueous solution was poured into the reaction mixture and extracted with ethyl acetate. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain (5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-6-fluoro- pyridin-2-carboxylic acid as a white solid form. The resulting product was used without an additional purification process.
[Step 4] Synthesis of tert-butyl 5-[2-fluoro-6-(isopropylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which 5-(2-tert-butoxycarbonyl-3,6-dihydrooxazin-5-yl)-6-fluoro- pyridin-2-carboxylic acid (100.00%, 0.200 g, 0.600 mmol), O-(7-azabenzotriazol-l-yl)- N,N,N',N'-tetramethyluronium hexafluorophosphate (100.00%, 0.352 g, 0.925 mmol), N- ethyldiisopropylamine (100.00% solution, 0.1612 mL, 0.925 mmol), and isopropylamine (100.00% solution, 0.05540 mL, 0.648 mmol) were dissolved in N,N-dimethylformamide (5 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated to obtain tert-butyl 5-[2-fluoro-6-(isopropylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (0.200 g, 90.00%) in a white solid form.
[Step 5] Synthesis of intermediate compound 13
A solution in which tert-butyl 5-[2-fluoro-6-(isopropylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.225 g, 0.616 mmol) and trifluoroacetic acid (100.00% solution, 0.4715 mL, 6.158 mmol) were dissolved in dichloromethane (10 mL) at room temperature was stirred at the same temperature for two hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 24 g cartridge; methanol/dichloromethane = 10 to 30%) and concentrated to obtain intermediate compound 13 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6-fluoro- N-isopropyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (0.150 g, 67.10%) in a white solid form.
Synthesis of intermediate compound 14: 5-(3,6-dihydro-2H-oxazin-5-yl)-6- methoxy-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of methyl 5-chloro-6-fluoro-pyridin-2-carboxylate
A solution in which methyl 5-chloropyridin-2-carboxylate (100.00%, 1.000 g, 5.828 mmol) and silver(II) fluoride (100.00%, 2.821 g, 19.340 mmol) were dissolved in acetonitrile (50 mL) at room temperature was stirred overnight at the same temperature. The reaction mixture was filtered via a celite pad to remove a solid therefrom, and then solvent was removed from the resulting filtrate without the solid under reduced pressure. Then, the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; ethyl acetate/hexane = 0 to 100%), and concentrated to obtain methyl 5-chloro-6-fluoro-pyridin-2- carboxylate (0.693 g, 62.72%) in a white solid form.
[Step 2] Synthesis of methyl 5-chloro-6-methoxy-pyridin-2-carboxylate
A solution in which methyl 5-chloro-6-fluoro-pyridin-2-carboxylate (100.00%, 1.000 g, 5.275 mmol), methanol (100.00% solution, 0.213 mL, 5.265 mmol), and potassium carbonate (100.00%, 1.046 g, 10.550 mmol) were dissolved in dimethylsulfoxide (3 mL) at 75°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; ethyl acetate/hexane = 0 to 100%), and concentrated to obtain methyl 5- chloro-6-methoxy-pyridin-2-carboxylate (0.950 g, 89.33%) in a white solid form.
[Step 3] Synthesis of 5-chloro-6-methoxy-N-methyl-pyridin-2-carboxamide
A reaction mixture in which methyl 5-chloro-6-methoxy-pyridin-2-carboxylate (100.00%, 0.950 g, 4.712 mmol) and methylamine solution (2.0 M solution in THF, 11.78 mL, 23.560 mmol) were mixed at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 12 g cartridge; ethyl acetate/hexane = 0 to 100%), and concentrated to obtain 5-chloro-6-methoxy-N-methyl- pyridin-2-carboxamide (0.945 g, 100.00%) in a white solid form.
[Step 4] Synthesis of tert-butyl 5-[2-methoxy-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which 5-chloro-6-methoxy-N-methyl-pyridin-2-carboxamide (100.00%, 0.150 g, 0.748 mmol), potassium carbonate (100.00%, 0.207 g, 1.495 mmol), [1,1'- bis(di-tert-butylphosphino)ferrocene]palladium(II) dichloride (100.00%, 0.049 g, 0.075 mmol), and tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydrooxazin-2- carboxylate (100.00%, 0.244 g, 0.785 mmol) were dissolved in 1,4-dioxane (20 mL)/water (4 mL) at 100°C was stirred at the same temperature for two hours, after which the temperature was lowered to room temperature to terminate the reaction. The reaction mixture was filtered via a celite pad to remove a solid therefrom, and then solvent was removed from the resulting filtrate without the solid under reduced pressure. Then, the resulting concentrate was purified via column chromatography (SiCT, 12 g cartridge; ethyl acetate/hexane = 0 to 100%) and concentrated to obtain tert-butyl 5-[2-methoxy-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (0.200 g, 76.56%) in a yellow solid form.
[Step 5] Synthesis of intermediate compound 14
A solution in which tert-butyl 5-[2-methoxy-6-(methylcarbamoyl)-3-pyridyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 0.184 g, 0.527 mmol) and trifluoroacetic acid (100.00% solution, 1.21 mL, 15.802 mmol) were dissolved in dichloromethane (50 mL) at room temperature was stirred at the same temperature for three hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 12 g cartridge; methanol/dichloromethane = 0 to 20%) and concentrated to obtain intermediate compound 14 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- methoxy-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (0.150 g, 82.01%) in a white solid form.
Synthesis of intermediate compound 15: 4-(3,6-dihydro-2H-oxazin-5-yl)-2- fluoro-N-methyl-benzamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-(3-fluoro-4-methoxycarbonyl-phenyl)-3,6- dihydrooxazin-2-carboxylate
A solution in which methyl 4-bromo-2-fluoro-benzoate (100.00%, 390.000 mg, 1.674 mmol), tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydrooxazin-2- carboxylate (100.00%, 547.000 mg, 1.758 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 109.000 mg, 0.167 mmol), and potassium carbonate (100.00%, 463.000 mg, 3.350 mmol) were dissolved in 1,4-dioxane (12 mL)/water (3 mL) at room temperature was stirred at 100°C for one hour, after which the temperature was lowered to room temperature to terminate the reaction. Water was poured into the reaction mixture and extracted with dichloromethane. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 12 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-(3-fluoro-4-methoxycarbonyl-phenyl)-3,6-dihydrooxazin- 2-carboxylate (545.000 mg, 96.54%) in a brown solid form.
[Step 2] Synthesis of tert-butyl 5-[3-fluoro-4-(methylcarbamoyl)phenyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(3-fluoro-4-methoxycarbonyl-phenyl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 545.000 mg, 1.616 mmol) and methylamine solution (2.0 M solution in THF, 1.6 mL, 3.200 mmol) were dissolved in methanol (5 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; ethyl acetate/hexane = 0 to 50%) and concentrated to obtain tert-butyl 5-[3-fluoro-4-(methylcarbamoyl)phenyl]-3,6-dihydrooxazin- 2-carboxylate (374.800 mg, 68.95%) in a white solid form.
[Step 3] Synthesis of intermediate compound 15 Boc
To a solution in which tert-butyl 5 - [3 -fluoro-4-(methylcarbamoyl)phenyl] -3 ,6- dihydrooxazin-2-carboxylate (100.00%, 1.410 g, 4.415 mmol) was dissolved in dichloromethane (10 mL) at room temperature, trifluoroacetic acid (100.00% solution, 10 mL, 130.680 mmol) was added and stirred at the same temperature for 30 minutes. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 40 g cartridge; methanol/dichloromethane = 5 to 30%) and concentrated to obtain intermediate compound 15 of 4-(3,6-dihydro-2H-oxazin- 5-yl)-2-fluoro-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (1.470 g, 99.91%) in a white solid form.
Synthesis of intermediate compound 16: 4-(3,6-dihydro-2H-oxazin-5-yl)-3- fluoro-N-methyl-benzamide; 2,2,2-trifluoroacetic acid
[Step 1] Synthesis of tert-butyl 5-(2-fluoro-4-methoxycarbonyl-phenyl)-3,6- dihydrooxazin-2-carboxylate
A solution in which methyl 4-bromo-3 -fluoro-benzoate (100.00%, 599.000 mg,
2.571 mmol), tert-butyl 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydrooxazin-2- carboxylate (100.00%, 840.000 mg, 2.699 mmol), [1, 1 '-bis(di-tert- butylphosphino)perrocene]palladium(II) dichloride (100.00%, 168.000 mg, 0.258 mmol), and potassium carbonate (100.00%, 711.000 mg, 5.145 mmol) were dissolved in 1,4-dioxane (8 mL)/water (2 mL) at room temperature was stirred at 100°C for one hour, after which the temperature was lowered to room temperature to terminate the reaction. Water was poured into the reaction mixture and extracted with dichloromethane. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 24 g cartridge; ethyl acetate/hexane = 0 to 30%) and concentrated to obtain tert-butyl 5-(2-fluoro-4-methoxycarbonyl-phenyl)-3,6-dihydrooxazin- 2-carboxylate (251.000 mg, 28.95%) in a brown solid form.
[Step 2] Synthesis of tert-butyl 5-[2-fluoro-4-(methylcarbamoyl)phenyl]-3,6- dihydrooxazin-2-carboxylate
A solution in which tert-butyl 5-(2-fluoro-4-methoxycarbonyl-phenyl)-3,6- dihydrooxazin-2-carboxylate (100.00%, 251.000 mg, 0.744 mmol) and methylamine solution (2.0 M solution in THF, 0.75 mL, 1.500 mmol) were dissolved in methanol (5 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; ethyl acetate/hexane = 0 to 50%) and concentrated to obtain tert-butyl 5-[2-fluoro-4-(methylcarbamoyl)phenyl]-3,6-dihydrooxazin- 2-carboxylate (158.000 mg, 63.12%) in a white solid form.
[Step 3] Synthesis of intermediate compound 16
To a solution in which tert-butyl 5-[2-fluoro-4-(methylcarbamoyl)phenyl]-3,6- dihydrooxazin-2-carboxylate (100.00%, 158.000 mg, 0.470 mmol) was dissolved in dichloromethane (3 mL) at room temperature, trifluoroacetic acid (100.00% solution, 1 mL, 13.068 mmol) was added and stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 40 g cartridge; methanol/dichloromethane = 5 to 30%) and concentrated to obtain intermediate compound 16 of 4-(3,6-dihydro-2H-oxazin-5- yl)-3-fluoro-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (135.000 mg, 82.05%) in a light yellow solid form.
Preparation of example compound
Synthesis of example compound 1, 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3- yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 100.000 mg, 0.374 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N- methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 131.000 mg, 0.393 mmol), and sodium hydrogen carbonate (100.00%, 94.000 mg, 1.119 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 25°C and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 1 of 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6- dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (65.100 mg, 42.89%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 5 11.85 (s, 1H), 8.72 - 8.71 (m, 1H), 8.65 - 8.64 (m, 1H), 8.44 - 8.43 (m, 1H), 7.98 - 7.97 (m, 2H), 7.75 - 7.74 (m, 1H), 7.69 - 7.68 (m, 1H), 6.63 - 6.62 (m, 1H), 4.68 (s, 2H), 4.09 - 4.07 (m, 2H), 3.60 - 3.59 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 2.54 (q, J = 6.1 Hz, 2H), 1.18 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 406.5 (M++ 1).
Synthesis of example compound 2, 5-[2-[(5-fluoro-2-methyl-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-8-fluoro-3-methyl-lH-quinoxalin-2-one (100.00%, 100.000 mg, 0.369 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 129.000 mg, 0.387 mmol), and sodium hydrogen carbonate (100.00%, 93.000 mg, 1.107 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 25°C and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 2 of 5-[2-[(5-fluoro-2-methyl-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6- dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (17.800 mg, 11.79%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.44 (s, 1H), 8.71 - 8.70 (m, 1H), 8.64 - 8.63 (m, 1H), 7.96 - 7.95 (m, 2H), 7.52 - 7.50 (m, 1H), 7.38 - 7.34 (m, 1H), 6.60 - 6.59 (m, 1H), 4.66 - 4.65 (m, 2H), 4.08 - 4.07 (m, 2H), 3.59 - 3.58 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 2.41 (s, 3H).; LRMS (ES) m/z 410.5 (M++ 1).
Synthesis of example compound 3, 5-[2-[(5-fluoro-2-isopropyl-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-8-fluoro-3-isopropyl-lH-quinoxalin-2-one (100.00%, 0.113 g, 0.378 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.120 g, 0.380 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1321 mL, 0.756 mmol) were dissolved in acetonitrile (15 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 3 of 5-[2-[(5-fluoro-2-isopropyl-3-oxo-4H-quinoxalin-6- yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.030 g, 18.00%) in a brown solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.46 (s, 1H), 8.71 (d, J = 5.0 Hz, 1H), 8.65 (s, 1H), 7.97 (d, J = 1.4 Hz, 2H), 7.56 - 7.54 (m, 1H), 7.38 (t, J = 3.7 Hz, 1H), 6.61 (s, 1H), 4.66 (s, 2H), 4.09 (s, 2H), 3.60 (s, 2H), 3.48 - 3.46 (m, 1H), 2.81 (d, J = 4.9 Hz, 3H), 1.22 (d, J = 6.8 Hz, 6H).; LRMS (ES) m/z 439.0 (M++ 1).
Synthesis of example compound 4, 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-quinoxalin-
6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.108 g, 0.378 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.120 g, 0.380 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1321 mL, 0.756 mmol) were dissolved in acetonitrile (15 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 4 of 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]- 3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.025 g, 16.00%) in a brown solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.46 (s, 1H), 8.71 (d, J = 5.0 Hz, 1H), 8.65 (s, 1H), 7.97 (d, J = 1.4 Hz, 2H), 7.56 - 7.54 (m, 1H), 7.39 - 7.37 (m, 1H), 6.61 (s, 1H), 4.66 (s, 2H), 4.09 (s, 2H), 3.60 (s, 2H), 3.48 - 3.46 (m, 1H), 2.81 (d, J = 4.9 Hz, 3H), 1.22 (d, J = 6.8 Hz, 6H).; LRMS (ES) m/z 424.2 (M++ 1).
Synthesis of example compound 5, 5-[2-[(2-ethyl-7-fluoro-3-oxo-4H-quinoxalin-
6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 2-ethyl-7-fluoro-3-oxo-4H-quinoxalin-6-carbaldehyde (100.00%, 0.070 g, 0.300 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.122 g, 0.366 mmol), and acetic acid (100.00% solution, 1 mL, 17.400 mmol) were dissolved in dichloromethane (30 mL) was stirred at room temperature for 15 minutes and sodium triacetoxyborohydride (100.00%, 0.287 g, 1.361 mmol) was added and further stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then water was poured into the resulting concentrate and extracted with dichloromethane. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated, and then the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 5 of 5-[2-[(2-ethyl- 7-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2- carboxamide (0.010 g, 7.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.31 (s, 1H), 8.72 (t, J = 4.8 Hz, 1H), 8.68 (s, 1H), 8.00 (s, 2H), 7.52 (dd, J = 20.4, 8.6 Hz, 2H), 6.66 (s, 1H), 4.74 (s, 2H), 4.08 (s, 2H), 3.64 (s, 2H), 2.84 - 2.78 (m, 5H), 1.23 - 1.22 (m, 3H).; LRMS (ES) m/z 425.2 (M++ 1).
Synthesis of example compound 6, 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3- yl)methyl]-3,6-dihydrooxazin-5-yl]-6-methoxy-N-methyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 0.115 g, 0.432 mmol), intermediate compound 14 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- methoxy-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.150 g, 0.430 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1508 mL, 0.864 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 6 of 5-[2-[(7-ethyl-6-oxo- 5H-l,5-naphthyridin-3-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-methoxy-N-methyl-pyridin-2- carboxamide (0.010 g, 5.30%) in a white solid form. 'H-NMR (400 MHz, DMSO-d6) 5 11.87 (s, 1H), 8.50 (d, J = 4.7 Hz, 1H), 8.43 (d, J = 1.5 Hz, 1H), 7.75 (t, J = 3.7 Hz, 2H), 7.69 (s, 1H), 7.59 (d, J = 7.6 Hz, 1H), 6.36 (s, 1H), 4.55 (s, 2H), 4.18 (s, 2H), 4.00 (s, 3H), 3.57 (s, 2H), 2.83 (d, J = 4.8 Hz, 3H), 2.56 - 2.54 (m, 2H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 436.4 (M+ + 1).
Synthesis of example compound 7, N-methyl-5-[2-[(2-methyl-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin-2-carboxamide
A solution in which 2-methyl-3-oxo-4H-l,4-benzoxazin-6-carbaldehyde (100.00%, 100.000 mg, 0.523 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N- methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 183.000 mg, 0.549 mmol), and sodium triacetoxyborohydride (100.00%, 333.000 mg, 1.571 mmol) were dissolved in dichloromethane (5 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; methanol/dichloromethane = 3%) and concentrated. Then, ethyl acetate (10 mL) was added into the resulting product and stirred to filter out a precipitated solid, which was then washed with ethyl acetate and dried to obtain example compound 7 of N-methyl-5-[2-[(2-methyl-3- oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin-2-carboxamide (80.100 mg, 38.83%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.61 (s, 1H), 8.71 - 8.70 (m, 1H), 8.65 - 8.64 (m, 1H), 7.97 - 7.96 (m, 2H), 6.95 - 6.94 (m, 1H), 6.90 - 6.89 (m, 2H), 6.59 - 6.58 (m, 1H), 4.66 - 4.65 (m, 3H), 3.85 - 3.84 (m, 2H), 3.49 - 3.48 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.41 (d, J = 6.8 Hz, 3H).; LRMS (ES) m/z 395.5 (M++ 1).
Synthesis of example compound 8, N-methyl-5-[2-[(3-methyl-2-oxo-lH- pyrido[2,3-b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin-2-carboxamide
To a solution in which 3-methyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7-carbaldehyde (100.00%, 100.000 mg, 0.520 mmol) and intermediate compound 1 of 5-(3,6-dihydro-2H- oxazin-5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 182.000 mg, 0.548 mmol) were dissolved in dichloromethane (5 mL) at room temperature, sodium triacetoxyborohydride (100.00%, 331.000 mg, 1.562 mmol) was added and stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated. Then, methanol (5 mL) was added into the resulting product and stirred to filter out a precipitated solid, which was then washed with ethyl acetate and dried to obtain example compound 8 of N-methyl-5-[2-[(3- methyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin-2- carboxamide (72.400 mg, 35.19%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.78 (s, 1H), 8.71 - 8.70 (m, 1H), 8.65 - 8.64 (m, 1H), 7.97 - 7.96 (m, 2H), 7.77 - 7.76 (m, 1H), 7.29 - 7.28 (m, 1H), 6.60 - 6.59 (m, 1H), 4.90 (q, J = 6.8 Hz, 1H), 4.65 (s, 2H), 3.89 - 3.88 (m, 2H), 3.53 - 3.52 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.47 (d, J = 6.8 Hz, 3H).; LRMS (ES) m/z 396.6 (M+ + 1).
Synthesis of example compound 9, 5-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 3-ethyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7-carbaldehyde (100.00%, 100.000 mg, 0.485 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 170.000 mg, 0.510 mmol), and sodium triacetoxyborohydride (100.00%, 308.000 mg, 1.453 mmol) were dissolved in dichloromethane (5 mL) at room temperature was stirred at the same temperature for 18 hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated. Then, methanol (5 mL) was added into the resulting product and stirred to filter out a precipitated solid, which was then washed with methanol and dried to obtain example compound 9 of 5-[2-[(3-ethyl-2-oxo-lH- pyrido[2,3-b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2- carboxamide (60.800 mg, 30.62%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.79 (s, 1H), 8.71 - 8.70 (m, 1H), 8.65 - 8.64 (m, 1H), 7.97 - 7.96 (m, 2H), 7.76 - 7.75 (m, 1H), 7.27 - 7.26 (m, 1H), 6.60 - 6.59 (m, 1H), 4.76 (m, 1H), 4.65 (s, 2H), 3.89 - 3.88 (m, 2H), 3.53 - 3.52 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.91 -1.89 (m. 1H), 1.84 -1.82 (m. 1H), 1.00 (d, J = 7.4 Hz, 3H).; LRMS (ES) m/z 410.6 (M+ + 1).
Synthesis of example compound 10, 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
To a solution in which 2-ethyl-5-fluoro-3-oxo-4H-l,4-benzoxazin-6-carbaldehyde (100.00%, 100.000 mg, 0.448 mmol) and intermediate compound 1 of 5-(3,6-dihydro-2H- oxazin-5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 157.000 mg, 0.471 mmol) were dissolved in dichloromethane (5 mL) at room temperature, sodium triacetoxyborohydride (100.00%, 285.000 mg, 1.345 mmol) was added and stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated. Then, methanol (10 mL) was added into the resulting product and stirred to filter out a precipitated solid, which was then washed with methanol and dried to obtain example compound 10 of 5-[2-[(2-ethyl-5- fluoro-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2- carboxamide (77.800 mg, 40.72%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.83 (s, 1H), 8.71 - 8.70 (m, 1H), 8.64 - 8.63 (m, 1H), 7.96 - 7.95 (m, 2H), 7.04 - 7.03 (m, 1H), 6.82 - 6.81 (m, 1H), 6.57 - 6.56 (m, 1H), 4.65 (s, 2H), 4.56 - 4.54 (m, 1H), 3.92 - 3.91 (m, 2H), 3.51 - 3.50 (m, 2H), 2.81 (d, J = 4.9 Hz, 3H), 1.85 -1.83 (m. 1H), 1.79 - 1.77 (m. 1H), 0.99 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 427.3 (M+ + 1).
Synthesis of example compound 11, 5-[2-[(5-fluoro-2-methyl-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
To a solution in which 6-(bromomethyl)-5-fluoro-2-methyl-4H-l,4-benzoxazin-3- one (100.00%, 100.000 mg, 0.365 mmol) and intermediate compound 1 of 5-(3,6-dihydro-2H- oxazin-5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 139.000 mg, 0.417 mmol) were dissolved in acetonitrile (5 mL) at room temperature, sodium hydrogen carbonate (100.00%, 75.000 mg, 0.893 mmol) was added and stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated. Then, methanol (10 mL) was added into the resulting product and stirred to filter out a precipitated solid, which was then washed with methanol and dried to obtain example compound 11 of 5-[2-[(5-fluoro-2-methyl-3-oxo-4H-l,4-benzoxazin-6- yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (43.500 mg, 28.91%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.83 (s, 1H), 8.71 - 8.70 (m, 1H), 8.64 - 8.63 (m, 1H), 7.96 - 7.95 (m, 2H), 7.04 - 7.03 (m, 1H), 6.82 - 6.81 (m, 1H), 6.57 - 6.56 (m, 1H), 4.71 - 4.70 (m, 1H), 4.65 (s, 2H), 3.92 - 3.91 (m, 2H), 3.52 - 3.51 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.43 (t, J = 6.8 Hz, 3H).; LRMS (ES) m/z 413.6 (M+ + 1).
Synthesis of example compound 12, 6-fluoro-N-methyl-5-[2-[(2-methyl-3-oxo- 4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin-2-carboxamide
A solution in which 2-methyl-3-oxo-4H-l,4-benzoxazin-6-carbaldehyde (100.00%, 28.000 mg, 0.146 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 48.300 mg, 0.138 mmol), and sodium triacetoxyborohydride (100.00%, 87.000 mg, 0.411 mmol) were dissolved in dichloromethane (2 mL) at room temperature was stirred at the same temperature for two hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; methanol/dichloromethane = 3%) and concentrated. Then, ethyl acetate (10 mL) was added into the resulting product and stirred to filter out a precipitated solid, which was then washed with ethyl acetate and dried to obtain example compound 12 of 6-fluoro-N-methyl-5-[2-[(2- methyl-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin-2- carboxamide (30.500 mg, 53.78%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.61 (s, 1H), 8.65 - 8.64 (m, 1H), 8.12 - 8.10 (m, 1H), 7.93 - 7.91 (m, 1H), 6.95 - 6.94 (m, 1H), 6.89 - 6.88 (m, 2H), 6.48 - 6.47 (m, 1H), 4.64 (q, J = 6.8 Hz, 1H), 4.57 - 4.56 (m, 2H), 3.85 (s, 2H), 3.50 - 3.49 (m, 2H), 2.79 (d, J = 4.8 Hz, 3H), 1.41 (t, J = 6.8 Hz, 3H).; LRMS (ES) m/z 413.6 (M+ + 1).
Synthesis of example compound 13, 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fhioro-N-methyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 0.040 g, 0.150 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6-fluoro- N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.050 g, 0.149 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.07 mL, 0.401 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 13 of 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6- dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2-carboxamide (0.020 g, 31.67%) in a yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 5 11.87 (s, 1H), 8.66 (d, J = 4.8 Hz, 1H), 8.43 (d, J = 1.7 Hz, 1H), 8.11 (q, J = 5.8 Hz, 1H), 7.93 (dd, J = 7.8, 1.7 Hz, 1H), 7.75 (s, 1H), 7.71 (s, 1H), 6.53 (s, 1H), 4.60 (s, 2H), 4.08 (s, 2H), 3.62 (s, 2H), 2.79 (d, J = 4.8 Hz, 3H), 2.56 - 2.54 (m, 2H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 424.4 (M+ + 1).
Synthesis of example compound 14, 6-fhioro-5-[2-[(5-fhioro-2-methyl-3-oxo-
4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-8-fluoro-3-methyl-lH-quinoxalin-2-one (100.00%, 0.054 g, 0.198 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.070 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.07 mL, 0.401 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 14 of 6-fluoro-5-[2-[(5-fluoro-2-methyl-3-oxo-4H-quinoxalin-6- yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.015 g, 20.00%) in a yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.45 (s, 1H), 8.65 (d, J = 4.9 Hz, 1H), 8.11 (t, J = 4.8 Hz, 1H), 8.07 (d, J = 2.8 Hz, 1H), 7.93 (d, J = 1.6 Hz, 1H), 7.48 (t, J = 37.7 Hz, 1H), 6.51 (s, 1H), 4.57 (s, 2H), 4.09 (s, 2H), 3.61 (s, 2H), 3.37 - 3.36 (m, 2H), 2.79 (d, J = 4.8 Hz, 3H), 2.42 (s, 3H).; LRMS (ES) m/z 428.2 (M++ 1).
Synthesis of example compound 15, 6-fhioro-5-[2-[(5-fhioro-2-methyl-3-oxo-
4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2- carboxamide
A solution in which 6-(bromomethyl)-5-fluoro-2-methyl-4H-l,4-benzoxazin-3-one (100.00%, 71.000 mg, 0.259 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 87.200 mg, 0.248 mmol), and sodium hydrogen carbonate (100.00%, 63.000 mg, 0.750 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; methanol/dichloromethane = 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 25°C and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 15 of 6-fluoro-5-[2-[(5-fluoro-2-methyl-3-oxo-4H-l,4-benzoxazin-6- yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (71.600 mg, 67.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.83 (s, 1H), 8.64 - 8.63 (m, 1H), 8.09 - 8.08 (m, 1H), 7.92 - 7.90 (m, 1H), 7.04 - 7.02 (m, 1H), 6.81 (d, J = 8.4 Hz, 1H), 6.48 - 6.47 (m, 1H), 4.70 (q, J = 6.7 Hz, 1H), 4.55 - 4.54 (m, 2H), 3.92 (s, 2H), 3.53 - 3.52 (m, 2H), 2.79 (d, J = 4.8 Hz, 3H), 1.43 (t, J = 6.7 Hz, 3H).; LRMS (ES) m/z 431.5 (M+ + 1).
Synthesis of example compound 16, 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide
A solution in which 2-ethyl-5-fluoro-3-oxo-4H-l,4-benzoxazin-6-carbaldehyde (100.00%, 28.000 mg, 0.125 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 48.300 mg, 0.138 mmol), and sodium triacetoxyborohydride (100.00%, 87.000 mg, 0.411 mmol) were dissolved in dichloromethane (2 mL) at room temperature was stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 3%) and concentrated. Then, methanol (10 mL) was added into the resulting product and stirred to filter out a precipitated solid, which was then washed with methanol and dried to obtain example compound 16 of 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2-carboxamide (30.500 mg, 49.90%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.83 (s, 1H), 8.64 - 8.63 (m, 1H), 8.11 - 8.09 (m, 1H), 7.92 - 7.90 (m, 1H), 7.04 - 7.02 (m, 1H), 6.82 (d, J = 8.3 Hz, 1H), 6.47 - 6.46 (m, 1H), 4.55 - 4.54 (m, 3H), 3.92 (s, 2H), 3.52 - 3.51 (m, 2H), 2.79 (d, J = 4.6 Hz, 3H), 1.83 - 1.79 (m, 2H), 0.99 (t, J = 7.3 Hz, 3H).; LRMS (ES) m/z 445.5 (M+ + 1).
Synthesis of example compound 17, 5-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fhioro-N-methyl-pyridin-2- carboxamide
A solution in which 3-ethyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7-carbaldehyde (100.00%, 49.000 mg, 0.238 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 80.000 mg, 0.228 mmol), and sodium triacetoxyborohydride (100.00%, 145.000 mg, 0.684 mmol) were dissolved in dichloromethane (2 mL) at room temperature was stirred at the same temperature for two hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 3%) and concentrated. Then, methanol (10 mL) was added into the resulting product and stirred to filter out a precipitated solid, which was then washed with methanol and dried to obtain example compound 17 of 5-[2-[(3-ethyl-2-oxo-lH- pyrido[2,3-b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide (29.400 mg, 30.20%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.80 (s, 1H), 8.65 - 8.64 (m, 1H), 8.12 - 8.10 (m, 1H), 7.93 - 7.91 (m, 1H), 7.76 (d, J = 1.8 Hz, 1H), 7.28 (d, J = 1.9 Hz, 1H), 6.49 - 6.48 (m, 1H), 4.77 - 4.76 (m, 1H), 4.56 - 4.55 (m, 2H), 3.89 (s, 2H), 3.54 - 3.53 (m, 2H), 2.79 (d, J = 4.8 Hz, 3H), 1.91 - 1.88 (m, 2H), 1.00 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 428.5 (M++ 1).
Synthesis of example compound 18, 6-fluoro-N-methyl-5-[2-[(3-methyl-2-oxo- lH-pyrido[2,3-b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin-2- carboxamide
A solution in which 3-methyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7-carbaldehyde (100.00%, 46.000 mg, 0.239 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 80.000 mg, 0.228 mmol), and sodium triacetoxyborohydride (100.00%, 145.000 mg, 0.684 mmol) were dissolved in dichloromethane (2 mL) at room temperature was stirred at the same temperature for two hours. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 3%) and concentrated. Then, methanol (10 mL) was added into the resulting product and stirred to filter out a precipitated solid, which was then washed with methanol and dried to obtain example compound 18 of 6-fluoro-N-methyl-5-[2- [(3-methyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin- 2-carboxamide (47.500 mg, 50.44%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.78 (s, 1H), 8.65 - 8.64 (m, 1H), 8.12 - 8.10 (m, 1H), 7.92 (dd, J = 7.7, 1.8 Hz, 1H), 7.76 (d, J = 2.0 Hz, 1H), 7.30 (d, J = 2.0 Hz, 1H), 6.49 - 6.48 (m, 1H), 4.90 (q, J = 6.8 Hz, 1H), 4.56 - 4.55 (m, 2H), 3.54 (s, 2H), 3.54 - 3.53 (m, 2H), 2.79 (d, J = 4.8 Hz, 3H), 1.47 (d, J = 6.8 Hz, 3H).; LRMS (ES) m/z 414.5 (M++ 1).
Synthesis of example compound 19, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.085 g, 0.298 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.100 g, 0.300 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.156 mL, 0.893 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 19 of 5-[2-[(2-ethyl-5-fluoro- 3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide (0.015 g, 10.00%) in a yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.44 (s, 1H), 8.65 (d, J = 4.7 Hz, 1H), 8.11 (t, J = 4.8 Hz, 1H), 7.92 (dd, J = 7.7, 1.7 Hz, 1H), 7.56 (d, J = 8.3 Hz, 1H), 7.37 (t, J = 7.6 Hz, 1H), 6.50 (s, 1H), 4.57 (s, 2H), 4.09 (s, 2H), 3.61 (s, 2H), 2.83 - 2.78 (m, 5H), 1.22 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 442.3 (M++ 1).
Synthesis of example compound 20, 5-[2-[(2-ethyl-7-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide
A solution in which 2-ethyl-7-fluoro-3-oxo-4H-quinoxalin-6-carbaldehyde
(100.00%, 0.100 g, 0.500 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.160 g, 0.455 mmol), and acetic acid (100.00% solution, 0.13 mL, 2.262 mmol) were dissolved in dichloromethane (20 mL) was stirred at room temperature for 15 minutes and sodium triacetoxyborohydride (100.00%, 0.287 g, 1.361 mmol) was added and further stirred overnight at the same temperature. Water was poured into the reaction mixture and extracted with dichloromethane. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated, and then the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 20 of 5-[2-[(2-ethyl- 7-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl- pyridin-2-carboxamide (0.031 g, 20.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.34 (s, 1H), 8.66 (d, J = 4.7 Hz, 1H), 8.13 (q, J = 5.9 Hz, 1H), 7.94 (dd, J = 7.8, 1.8 Hz, 1H), 7.55 (d, J = 10.5 Hz, 1H), 7.50 (d, J = 6.9 Hz, 1H), 6.54 (s, 1H), 4.66 (s, 2H), 4.08 (s, 2H), 3.65 (s, 2H), 2.82 - 2.79 (m, 5H), 1.22 (t, J = 14.3 Hz, 3H).; LRMS (ES) m/z 442.3 (M++ 1).
Synthesis of example compound 21, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-methoxy-N-methyl-pyridin-2- carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.051 g, 0.178 mmol), intermediate compound 14 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-methoxy-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.065 g, 0.190 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.09805 mL, 0.561 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 21 of 5-[2-[(2-ethyl-5-fluoro- 3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-methoxy-N-methyl-pyridin-2- carboxamide (0.005 g, 5.90%) in a white solid form.
'H-NMR (400 MHz, CDC13) 6 9.20 (s, 1H), 7.78 (d, J = 7.5 Hz, 1H), 7.70 (d, J = 4.9 Hz, 1H), 7.63 (t, J = 15.2 Hz, 2H), 7.45 (t, J = 7.9 Hz, 1H), 6.19 (t, J = 1.7 Hz, 1H), 4.64 (s, 2H), 4.11 (s, 2H), 3.99 (s, 3H), 3.61 (t, J = 1.7 Hz, 2H), 3.04 - 2.97 (m, 5H), 1.34 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 454.1 (M++ 1).
Synthesis of example compound 22, N-ethyl-5-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fhioro-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-pyrido[2,3-b][l,4]oxazine-2-one (100.00%, 52.000 mg, 0.192 mmol), intermediate compound 11 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-ethyl-6-fluoro-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 64.000 mg, 0.175 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.153 mL, 0.878 mmol) were dissolved in acetonitrile (3 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was crystallized with methanol (10 mL) at room temperature and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 22 of N -ethyl-5- [2- [(3 -ethyl-2-oxo- 1 H-pyrido [2,3 -b] [ 1 ,4] oxazin-7 -yl)methyl] -3 ,6-dihydrooxazin- 5-yl]-6-fluoro-pyridin-2-carboxamide (56.700 mg, 73.30%) in an ivory solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.78 (s, 1H), 8.69 (t, J = 5.9 Hz, 1H), 8.12 - 8.10 (m, 1H), 7.93 - 7.91 (m, 1H), 7.76 (d, J = 1.6 Hz, 1H), 7.28 (d, J = 1.7 Hz, 1H), 6.49 - 6.48 (m, 1H), 4.75 - 4.74 (m, 1H), 4.56 - 4.55 (m, 2H), 3.89 (s, 2H), 3.54 - 3.53 (m ,2H), 3.30 - 3.27 (m ,2H), 1.89 - 1.82 (m ,2H), 1.10 (t, J = 7.1 Hz, 3H), 0.99 (t, J = 7.3 Hz, 3H).; LRMS (ES) m/z 442.6 (M+ + 1).
Synthesis of example compound 23, N-ethyl-5-[2-[(2-ethyl-5-fluoro-3-oxo-4Hl,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fhioro-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-5-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 56.000 mg, 0.194 mmol), intermediate compound 11 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-ethyl-6-fluoro-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 64.000 mg, 0.175 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.153 mL, 0.878 mmol) were dissolved in acetonitrile (3 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was crystallized with methanol (10 mL) at room temperature and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 23 of N -ethyl-5- [2- [(2-ethyl-5-fluoro-3 -oxo-4H- 1 ,4-benzoxazin-6-yl)methyl] -3 ,6-dihydrooxazin-5 - yl]-6-fluoro-pyridin-2-carboxamide (42.100 mg, 52.41%) in an ivory solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.83 (s, 1H), 8.69 (t, J = 5.9 Hz, 1H), 8.10 - 8.08 (m, 1H), 7.92 - 7.90 (m, 1H), 7.04 - 7.02 (m, 1H), 6.82 - 6.80 (m, 1H), 6.47 - 6.46 (m, 1H), 4.55 - 4.54 (m, 2H), 3.92 (s, 2H), 3.52 - 3.51 (m, 2H), 3.30 - 3.28 (m, 2H), 1.84 - 1.77 (m, 2H), 1.10 (t, J = 7.1 Hz, 3H), 0.99 (t, J = 7.3 Hz, 3H).; LRMS (ES) m/z 459.6 (M++ 1).
Synthesis of example compound 24, N-ethyl-5-[2-[(7-ethyl-6-oxo-5H-l,5- naphthyridin-3-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-pyridin-2-carboxamide A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 51.000 mg, 0.191 mmol), intermediate compound 11 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N- ethyl-6-fluoro-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 64.000 mg, 0.175 mmol), and N,N-diisopropylethylamine (100.00% solution, 153 mL, 878.391 mmol) were dissolved in acetonitrile (3 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was crystallized with methanol (10 mL) at room temperature and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 24 of N-ethyl- 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro- pyridin-2-carboxamide (15.900 mg, 20.74%) in a brown solid form.
'H-NMR (400 MHz, DMSO-d6) 6 11.87 (s, 1H), 8.70 - 8.69 (m, 1H), 8.42 - 8.41 (m, 1H), 8.11 - 8.10 (m, 1H), 7.94 - 7.93 (m, 1H), 7.75 - 7.70 (m, 2H), 6.52 - 6.51 (m, 1H), 4.59 - 4.58 (m, 2H), 4.07 - 4.06 (m, 2H), 3.62 - 3.61 (m, 2H), 3.34 - 3.33 (m, 2H), 2.50 - 2.49 (m, 2H), 1.18 - 1.17 (m, 3H), 1.10 - 1.09 (m, 3H).; LRMS (ES) m/z 438.5 (M+ + 1).
Synthesis of example compound 25, N-ethyl-5-[2-[(2-ethyl-5-fluoro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.111 g, 0.389 mmol), intermediate compound 11 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-ethyl-6-fluoro-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.136 g, 0.389 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.2041 mL, 1.168 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 25 of N-ethyl-5-[2-[(2-ethyl- 5-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-pyridin-2- carboxamide (0.018 g, 10.10%) in a white solid form.
'H-NMR (400 MHz, CDC13) 6 9.26 (s, 1H), 8.06 (dd, J = 7.7, 1.8 Hz, 1H), 7.81 (dd, J = 9.3, 7.7 Hz, 1H), 7.64 - 7.62 (m, 2H), 7.44 (t, J = 7.9 Hz, 1H), 6.39 - 6.37 (m, 1H), 4.63 (s, 2H), 4.12 (s, 2H), 3.63 (t, J = 1.7 Hz, 2H), 3.51 - 3.47 (m, 2H), 2.99 - 2.97 (m, 2H), 1.35 (t, J = 7.4 Hz, 3H), 1.26 (t, J = 7.3 Hz, 3H).; LRMS (ES) m/z 456.2 (M+ + 1).
Synthesis of example compound 26, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-isopropyl-pyridin-2- carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.118 g, 0.413 mmol), intermediate compound 13 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-fluoro-N-isopropyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.150 g, 0.410 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.2164 mL, 1.239 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 26 of 5-[2-[(2-ethyl-5-fluoro- 3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-isopropyl-pyridin-2- carboxamide (0.018 g, 9.30%) in a white solid form.
'H-NMR (400 MHz, CDC13) 6 9.38 (s, 1H), 8.06 (dd, J = 7.7, 1.7 Hz, 1H), 7.80 (d, J = 17.0 Hz, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.45 (q, J = 7.2 Hz, 2H), 6.37 (s, 1H), 4.63 (s, 2H), 4.28 - 4.26 (m, 1H), 4.12 (s, 2H), 3.63 (s, 2H), 2.99 - 2.98 (m, 2H), 1.33 (t, J = 24.6 Hz, 3H), 1.27 (d, J = 6.6 Hz, 6H).; LRMS (ES) m/z 470.2 (M++ 1).
Synthesis of example compound 27, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-propyl-pyridin-2- carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.078 g, 0.275 mmol), intermediate compound 12 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-fluoro-N-propyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.100 g, 0.300 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1442 mL, 0.826 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 27 of 5-[2-[(2-ethyl-5-fluoro- 3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-propyl-pyridin-2- carboxamide (0.008 g, 6.00%) in a white solid form.
'H-NMR (400 MHz, CDC13) 6 9.13 (s, 1H), 8.07 (dd, J = 7.3, 1.3 Hz, 1H), 7.81 (t, J = 4.6 Hz, 1H), 7.67 - 7.62 (m, 2H), 7.44 (t, J = 7.9 Hz, 1H), 6.38 (s, 1H), 4.63 (s, 2H), 4.12 (s, 2H), 3.63 (d, J = 3.4 Hz, 2H), 3.41 (q, J = 6.8 Hz, 2H), 2.98 (q, J = 7.4 Hz, 2H), 1.67 - 1.69 (m, 2H), 1.35 (t, J = 7.4 Hz, 3H), 0.99 (t, J = 3.7 Hz, 3H).; LRMS (ES) m/z 470.2 (M++l).
Synthesis of example compound 28, 5-[2-[(2-ethyl-7-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-methoxy-N-methyl-pyridin-2- carboxamide
A solution in which 2-ethyl-7-fluoro-3-oxo-4H-quinoxalin-6-carbaldehyde
(100.00%, 0.100 g, 0.500 mmol), intermediate compound 14 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-methoxy-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.160 g, 0.440 mmol), and acetic acid (100.00% solution, 1 mL, 17.400 mmol) were dissolved in dichloromethane (15 mL) was stirred at room temperature for 15 minutes and sodium triacetoxyborohydride (100.00%, 0.289 g, 1.362 mmol) was added and further stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 28 of 5-[2-[(2-ethyl- 7-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-methoxy-N-methyl- pyridin-2-carboxamide (0.005 g, 2.00%) in a white solid form.
'H-NMR (400 MHz, CDC13) 6 10.69 (s, 1H), 7.81 (d, J = 7.6 Hz, 1H), 7.71 (d, J = 2.9 Hz, 1H), 7.62 (d, J = 7.4 Hz, 1H), 7.37 (d, J = 130.0 Hz, 1H), 7.43 (d, J = 6.2 Hz, 1H), 6.22 (s, 1H), 4.70 (s, 2H), 4.11 (s, 2H), 3.97 (s, 3H), 3.64 (s, 2H), 3.05 (d, J = 4.9 Hz, 3H), 2.94 (q, J = 3.8 Hz, 2H), 1.28 (t, J = 7.3 Hz, 3H).; LRMS (ES) m/z 455.4 (M+ + 1).
Synthesis of example compound 29, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N,3-dimethyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.043 g, 0.151 mmol), intermediate compound 5 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N,3-dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.050 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.07907 mL, 0.453 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 29 of 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]- 3,6-dihydrooxazin-5-yl]-N,3-dimethyl-pyridin-2-carboxamide (0.010 g, 20.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.44 (s, 1H), 8.53 (d, J = 5.3 Hz, 1H), 8.46 (d, J = 1.7 Hz, 1H), 7.74 (s, 1H), 7.55 (d, J = 8.2 Hz, 1H), 7.38 (t, J = 3.9 Hz, 1H), 6.57 (s, 1H), 4.65 (s, 2H), 4.09 (s, 2H), 3.59 (s, 2H), 2.83 - 2.76 (m, 5H), 2.54 (s, 3H), 1.22 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 438.3 (M++ 1).
Synthesis of example compound 30, 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-N,3-dimethyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 0.040 g, 0.151 mmol), intermediate compound 5 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N,3- dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.050 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.07907 mL, 0.453 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 30 of 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6- dihydrooxazin-5-yl]-N,3-dimethyl-pyridin-2-carboxamide (0.010 g, 20.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 5 11.85 (s, 1H), 8.53 (d, J = 4.9 Hz, 1H), 8.56 (d, J = 104.1 Hz, 2H), 7.75 (d, J = 5.3 Hz, 2H), 7.69 (s, 1H), 6.59 (s, 1H), 4.66 (s, 2H), 4.07 (s, 2H), 3.60 (s, 2H), 2.77 (d, J = 4.7 Hz, 3H), 2.63 - 2.54 (m, 5H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 420.4 (M+ + 1).
Synthesis of example compound 31, 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin- 3-yl)methyl]-3,6-dihydrooxazin-5-yl]-N,4-dimethyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 0.048 g, 0.181 mmol), intermediate compound 9 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N,4- dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.09489 mL, 0.543 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 10%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 31 of 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6- dihydrooxazin-5-yl]-N,4-dimethyl-pyridin-2-carboxamide (0.010 g, 10.00%) in a white solid form. 'H-NMR (400 MHz, DMSO-d6) 8 11.85 (s, 1H), 8.76 (d, J = 1.9 Hz, 1H), 8.56 (d, J = 4.5 Hz, 1H), 8.43 (d, J = 1.6 Hz, 1H), 8.04 (d, J = 1.7 Hz, 1H), 7.75 (d, J = 2.5 Hz, 2H), 6.30 (s, 1H), 4.63 (s, 2H), 4.09 (s, 2H), 3.61 (s, 2H), 2.79 (d, J = 4.5 Hz, 3H), 2.55 - 2.46 (m, 5H), 1.18 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 421.3 (M+ + 1).
Synthesis of example compound 32, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N,4-dimethyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.052 g, 0.181 mmol), intermediate compound 9 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N,4-dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.09489 mL, 0.543 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 32 of 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]- 3,6-dihydrooxazin-5-yl]-N,4-dimethyl-pyridin-2-carboxamide (0.010 g, 10.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.44 (s, 1H), 8.75 (d, J = 1.9 Hz, 1H), 8.55 (d, J = 4.6 Hz, 1H), 8.03 (d, J = 1.7 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.39 (t, J = 7.7 Hz, 1H), 6.28 (s, 1H), 4.59 (s, 2H), 4.11 (s, 2H), 3.59 (s, 2H), 2.83 - 2.78 (m, 5H), 2.44 (s, 3H), 1.22 (t, J =
7.4 Hz, 3H).; LRMS (ES) m/z 438.1 (M++ 1).
Synthesis of example compound 33, 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N,3-dimethyl-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-5-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 0.043 g, 0.149 mmol), intermediate compound 5 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N,3-dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.050 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.07907 mL, 0.453 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 33 of 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4-benzoxazin-6- yl)methyl]-3,6-dihydrooxazin-5-yl]-N,3-dimethyl-pyridin-2-carboxamide (0.010 g, 20.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.84 (s, 1H), 8.53 (d, J = 4.2 Hz, 1H), 8.45 (d, J = 1.7 Hz, 1H), 7.73 (s, 1H), 7.03 (t, J = 8.1 Hz, 1H), 6.82 (d, J = 8.3 Hz, 1H), 6.54 (s, 1H), 4.63 (s, 2H), 4.55 (q, J = 4.1 Hz, 1H), 3.92 (s, 2H), 3.51 (s, 2H), 2.76 (d, J = 4.8 Hz, 3H), 2.54 (s, 3H), 1.86 - 1.74 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 441.2 (M++ 1). Synthesis of example compound 34, 5-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-N,3-dimethyl-pyridin-2- carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-pyrido[2,3-b][l,4]oxazine-2-one (100.00%, 0.043 g, 0.159 mmol), intermediate compound 5 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N,3-dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.050 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.07907 mL, 0.453 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 34 of 5-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7- yl)methyl]-3,6-dihydrooxazin-5-yl]-N,3-dimethyl-pyridin-2-carboxamide (0.010 g, 20.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.80 (s, 1H), 8.53 (d, J = 4.9 Hz, 1H), 8.46 (d, J = 2.0 Hz, 1H), 7.75 (d, J = 12.8 Hz, 2H), 7.27 (d, J = 2.0 Hz, 1H), 6.56 (s, 1H), 4.76 (q, J = 4.0 Hz, 1H), 4.64 (s, 2H), 3.89 (s, 2H), 3.52 (s, 2H), 2.77 (d, J = 4.8 Hz, 3H), 2.54 (s, 3H), 1.90 - 1.79 (m, 2H), 1.00 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 424.3 (M+ + 1). Synthesis of example compound 35, 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyrimidin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 53.000 mg, 0.198 mmol), intermediate 4 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N-methyl- pyrimidin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 60.000 mg, 0.180 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.156 mL, 0.896 mmol) were dissolved in acetonitrile (3 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 3 to 5%) and concentrated. Then, the resulting product was crystallized with methanol (1 mL) at 25°C and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 35 of 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6- dihydrooxazin-5-yl]-N-methyl-pyrimidin-2-carboxamide (12.900 mg, 17.68%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 11.95 (s, 1H), 8.97 (s, 2H), 8.84 - 8.83 (m, 1H), 8.47 (s, 1H), 7.76 - 7.75 (m, 2H), 6.76 - 6.75 (m, 1H), 4.70 - 4.69 (m, 2H), 4.09 - 4.08 (m, 2H), 3.63 - 3.62 (m, 2H), 2.81 (d, J = 4.7 Hz, 3H), 2.56 - 2.54 (m, 2H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 407.5 (M++ 1).
Synthesis of example compound 36, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyrimidin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 72.000 mg, 0.253 mmol), intermediate compound 4 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyrimidin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 80.000 mg, 0.239 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.2 mL, 1.148 mmol) were dissolved in acetonitrile (3 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 3 to 5%) and concentrated. Then, the resulting product was crystallized with methanol (1 mL) at 25°C and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 36 of 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6- dihydrooxazin-5-yl]-N-methyl-pyrimidin-2-carboxamide (9.700 mg, 9.55%) in a yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.44 (s, 1H), 8.95 (s, 2H), 8.83 - 8.82 (m, 1H), 7.55 - 7.54 (m, 1H), 7.36 - 7.35 (m, 1H), 6.73 - 6.72 (m, 1H), 4.68 - 4.67 (m, 2H), 4.09 - 4.08 (m, 2H), 3.61 - 3.60 (m, 2H), 2.81 - 2.80 (m, 5H), 1.22 - 1.21 (m, 3H).; LRMS (ES) m/z 425.5 (M++ 1).
Synthesis of example compound 37, 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyrimidin-2-carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-5-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 72.000 mg, 0.250 mmol), intermediate compound 4 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyrimidin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 80.000 mg, 0.239 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.2 mL, 1.148 mmol) were dissolved in acetonitrile (3 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 3 to 5%) and concentrated. Then, the resulting product was crystallized with methanol (1 mL) at 25°C and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 37 of 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6- dihydrooxazin-5-yl]-N-methyl-pyrimidin-2-carboxamide (11.500 mg, 11.24%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.84 (s, 1H), 8.95 (s, 2H), 8.83 - 8.82 (m, 1H), 7.04 - 7.02 (m, 1H), 6.82 - 6.80 (m, 1H), 6.70 - 6.69 (m, 1H), 4.67 - 4.66 (m, 2H), 4.56 - 4.55 (m, 1H), 3.92 - 3.91 (m, 2H), 3.53 - 3.52 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.86 - 1.84 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 428.5 (M+ + 1).
Synthesis of example compound 38, 5-[2-[(7-fluoro-2-methyl-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-7-fluoro-2-methyl-4H-l,4-benzoxazin-3-one (100.00%, 100.000 mg, 0.365 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 128.000 mg, 0.384 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.3 mL, 1.722 mmol) were dissolved in acetonitrile (5 mL) at room temperature was added and stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then a precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 38 of 5-[2- [(7-fluoro-2-methyl-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N- methyl-pyridin-2-carboxamide (108.500 mg, 72.11%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.63 (s, 1H), 8.72 (q, J = 4.5 Hz, 1H), 8.65 - 8.64 (m, 1H), 7.98 - 7.97 (m, 2H), 7.05 (d, J = 7.3 Hz, 1H), 6.88 - 6.87 (m, 1H), 6.60 - 6.59 (m, 1H), 4.70 - 4.68 (m, 3H), 3.89 - 3.88 (m, 2H), 3.54 - 3.53 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.42 (d, J = 6.8 Hz, 3H).; LRMS (ES) m/z 413.5 (M+ + 1).; LRMS (ES) m/z 413.5 (M++ 1).
Synthesis of example compound 39, 5-[2-[(2-ethyl-7-fluoro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-7-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 100.000 mg, 0.347 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 121.000 mg, 0.363 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.3 mL, 1.722 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 39 of 5- [2- [(2-ethyl-7 -fluoro-3 -oxo-4H- 1 ,4-benzoxazin-6-yl)methyl] -3 ,6-dihydrooxazin-5-yl] -N- methyl-pyridin-2-carboxamide (96.900 mg, 65.47%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.64 (s, 1H), 8.72 - 8.71 (m, 1H), 8.65 - 8.64 (m ,1H), 7.98 - 7.97 (m, 2H), 7.03 (d, J = 7.3 Hz, 1H), 6.89 - 6.86 (m, 1H), 6.60 - 6.59 (m, 1H), 4.68 - 4.67 (m, 2H), 4.55 - 4.54 (m, 1H), 3.89 - 3.88 (m, 2H), 3.53 - 3.52 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.83 - 1.78 (m, 2H), 0.98 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 427.5 (M+ + 1).
Synthesis of example compound 40, 5-[2-[(8-fluoro-2-methyl-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-8-fluoro-2-methyl-4H-l,4-benzoxazin-3-one (100.00%, 100.000 mg, 0.365 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 128.000 mg, 0.384 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.3 mL, 1.722 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 40 of 5- [2- [(8-fluoro-2-methyl-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N- methyl-pyridin-2-carboxamide (96.800 mg, 64.34%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.82 (s, 1H), 8.72 - 8.70 (m, 1H), 8.65 - 8.64 (m, 1H), 7.97 - 7.96 (m, 2H), 6.89 - 6.88 (m, 1H), 6.78 -6.77 (m, 1H), 6.60 - 6.59 (m, 1H), 4.76 (q, J = 6.7 Hz, 1H), 4.67 - 4.66 (m, 2H), 3.85 (s, 2H), 3.51 - 3.50 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.45 (d, J = 6.8 Hz, 3H).; LRMS (ES) m/z 413.5 (M++ 1).
Synthesis of example compound 41, 5-[2-[(2-ethyl-8-fluoro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-8-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 100.000 mg, 0.347 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 121.000 mg, 0.363 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.3 mL, 1.722 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 41 of 5-[2- [(2-ethyl-8-fluoro-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N- methyl-pyridin-2-carboxamide (97.300 mg, 65.74%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.82 (s, 1H), 8.72 - 8.71 (m, 1H), 8.65 - 8.64 (m, 1H), 7.98 - 7.97 (m, 2H), 6.89 - 6.86 (m, 1H), 6.77 - 6.76 (m, 1H), 6.60 - 6.59 (m, 1H), 4.67 - 4.66 (m, 2H), 1.62 - 4.60 (m, 1H), 3.85 (s, 2H), 3.51 - 3.50 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.87 - 1.82 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 427.5 (M+ + 1). Synthesis of example compound 42, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-3-(trifluoromethyl)pyridin-2- carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-pyrido[2,3-b][l,4]oxazine-2-one (100.00%, 0.043 g, 0.159 mmol), intermediate compound 6 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-3-(trifluoromethyl)pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.050 g, 0.100 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.07907 mL, 0.453 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 42 of 5-[2-[(2-ethyl-5-fluoro- 3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-3- (trifluoromethyl)pyridin-2-carboxamide (0.010 g, 20.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 8 12.45 (s, 1H), 8.85 (d, J = 1.8 Hz, 1H), 8.60 (d, J = 4.8 Hz, 1H), 8.21 (d, J = 1.7 Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.37 (t, J = 2.4 Hz, 1H), 6.73 (s, 1H), 4.71 (s, 2H), 4.10 (s, 2H), 3.61 (s, 2H), 2.83 - 2.77 (m, 5H), 1.22 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 474.1 (M++ 1). Synthesis of example compound 43, 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-3-(trifhioromethyl)pyridin-2- carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 0.048 g, 0.181 mmol), intermediate compound 6 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N-methyl- 3-(trifluoromethyl)pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.09489 mL, 0.543 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 43 of 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6- dihydrooxazin-5-yl]-N-methyl-3-(trifluoromethyl)pyridin-2-carboxamide (0.010 g, 10.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 8 11.87 (s, 1H), 8.87 (d, J = 1.8 Hz, 1H), 8.60 (d, J = 4.9 Hz, 1H), 8.44 (d, J = 1.8 Hz, 1H), 8.21 (d, J = 1.9 Hz, 1H), 7.76 (s, 1H), 7.69 (d, J = 1.3 Hz, 1H), 6..7591 (s, 1H), 4.72 (s, 2H), 4.08 (s, 2H), 3.62 (s, 2H), 2.78 (d, J = 4.7 Hz, 3H), 2.55 (q, J = 2.5 Hz, 2H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 474.1 (M+ + 1).
Synthesis of example compound 44, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N,4-dimethyl-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-5-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 0.064 g, 0.222 mmol), intermediate compound 9 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N,4-dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.070 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1107 mL, 0.634 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 5%) and concentrated. Then, methanol (10 mL) and dichloromethane (15 mL) were added into the resulting product and stirred to filter out a precipitated solid, which was then washed with methanol and dried to obtain example compound 44 of 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H- 1 ,4-benzoxazin-6-yl)methyl] -3 ,6-dihydrooxazin-5 -yl] -N,4-dimethyl-pyridin-2-carboxamide (0.005 g, 5.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.84 (s, 1H), 8.75 (d, J = 1.8 Hz, 1H), 8.55 (d, J = 4.7 Hz, 1H), 8.02 (d, J = 1.9 Hz, 1H), 7.04 (t, J = 0.6 Hz, 1H), 6.82 (d, J = 8.4 Hz, 1H), 6.25 (s, 1H), 4.57 - 4.53 (m, 3H), 3.94 (s, 2H), 3.51 (s, 2H), 2.79 (d, J = 4.5 Hz, 3H), 2.43 (s, 3H), 1.83 - 1.74 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 441.1 (M+ + 1).
Synthesis of example compound 45, 5-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyrimidin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-pyrido[2,3-b][l,4]oxazine-2-one
(100.00%, 68.000 mg, 0.251 mmol), intermediate compound 4 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyrimidin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 80.000 mg, 0.239 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.2 mL, 1.148 mmol) were dissolved in acetonitrile (3 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then a precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 45 of 5- [2- [(3- ethyl-2-oxo- 1 H-pyrido [2 ,3 -b] [ 1 ,4]oxazin-7 -yl)methyl] -3 ,6-dihydrooxazin-5-yl] -N-methyl- pyrimidin-2-carboxamide (15.900 mg, 16.19%) in a brown solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.81 (s, 1H), 8.97 - 8.95 (m, 2H), 8.84 - 8.83 (m, 1H), 7.76 - 7.75 (m, 1H), 7.27 - 7.26 (m, 1H), 6.72 - 6.71 (m, 1H), 4.76 - 4.75 (m, 1H), 4.67 -
4.66 (m, 2H), 3.89 - 3.88 (m, 2H), 3.54 - 3.53 (m, 2H), 2.81 (d, J = 4.8 Hz, 3H), 1.89 - 1.85 (m,
2H), 0.99 (t, J = 7.3 Hz, 3H).; LRMS (ES) m/z 411.5 (M+ + 1).
Synthesis of example compound 46, 5-[2-[(5-fluoro-2,2-dimethyl-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-5-fluoro-2,2-dimethyl-4H-l,4-benzoxazin-3- one (100.00%, 0.050 g, 0.174 mmol), intermediate compound 1 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 1.000 equiv., 0.174 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.15114 mL, 0.868 mmol) were dissolved in acetonitrile (10 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; methanol/dichloromethane = 0 to 20%) and concentrated to obtain example compound 46 of 5 - [2- [(5-fluoro-2,2-dimethyl-3 -oxo-4H- 1 ,4-benzoxazin-6-yl)methyl] -3 ,6-dihydrooxazin-5- yl]-N-methyl-pyridin-2-carboxamide (0.053 g, 71.62%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.79 (s, 1H), 8.79 - 8.69 (m, 1H), 8.63 - 8.62 (m, 1H), 7.96 - 7.95 (m, 2H), 7.04 - 7.00 (m, 1H), 6.78 - 6.76 (m, 1H), 6.57 (s, 1H), 4.64 (s, 2H), 3.92 (s, 2H), 3.47 - 3.38 (m, 2H), 2.81 (d, J = 4.88Hz, 3H), 1.40 (s, 6H).; LRMS (ES) m/z 427.3 (M++ 1).
Synthesis of example compound 47, 6-fluoro-5-[2-[(5-fhioro-2,2-dimethyl-3- oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2- carboxamide
A solution in which 6-(bromomethyl)-5-fluoro-2,2-dimethyl-4H-l,4-benzoxazin-3- one (100.00%, 0.050 g, 0.174 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin- 5-yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 1.000 equiv., 0.174 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.15114 mL, 0.868 mmol) were dissolved in acetonitrile (10 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; methanol/dichloromethane = 0 to 20%) and concentrated to obtain example compound 47 of 6-fluoro-5-[2-[(5-fluoro-2,2-dimethyl-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6- dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.053 g, 71.62%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.78 (s, 1H), 8.65 - 8.62 (m, 1H), 8.10 - 8.05 (m, 1H), 7.92 - 7.89 (m, 1H), 7.04 - 7.00 (m, 1H), 6.79 - 6.77 (m, 1H), 6.47 (s, 1H), 4.55 (s, 2H), 3.91 (s, 2H), 3.47 - 3.38 (m, 2H), 2.79 (d, J = 4.84, 3H), 1.40 (s, 6H).; LRMS (ES) m/z 445.2 (M++ 1).
Synthesis of example compound 48, 5-[2-[(5-fhioro-3-oxo-spiro[4H-l,4- benzoxazin-2,l'-cyclobutene]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2- carboxamide
A solution in which 6-(bromomethyl)-5-fluoro-spiro[4H-l,4-benzoxazin-2,l'- cyclobutane]-3-one (100.00%, 0.050 g, 0.167 mmol), intermediate compound 1 of 5-(3,6- dihydro-2H-oxazin-5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 1.000 equiv., 0.167 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.14509 mL, 0.833 mmol) were dissolved in acetonitrile (10 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 48 of 5-[2-[(5-fluoro-3-oxo-spiro[4H-l,4-benzoxazin-2,l'- cyclobutene]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.050 g, 68.46%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.82 (s, 1H), 8.71 - 8.69 (m, 1H), 8.64 - 8.63 (m, 1H), 7.96 - 7.96 (m, 2H), 7.05 - 7.01 (m, 1H), 6.86 - 6.84 (m, 1H), 6.58 (s, 1H), 4.65 (s, 2H), 3.92 (s, 2H), 3.52 (s, 2H), 2.81 (d, j = 4.88 Hz, 3H), 2.53 - 2.47 (m, 2H), 2.27 - 2.21 (m, 2H), 1.96 - 1.88 (m, 1H), 1.81 - 1.77 (m, 1H).; LRMS (ES) m/z 439.3 (M++ 1).
Synthesis of example compound 49, 6-fhioro-5-[2-[(5-fhioro-3-oxo-spiro[4H- l,4-benzoxazin-2,l'-cyclobutene]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl- pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-5-fluoro-spiro[4H-l,4-benzoxazin-2,r- cyclobutane]-3-one (100.00%, 0.050 g, 0.167 mmol), intermediate compound 2 of 5-(3,6- dihydro-2H-oxazin-5-yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 1.050 equiv., 0.175 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.14509 mL, 0.833 mmol) were dissolved in acetonitrile (10 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 49 of 6-fluoro-5-[2-[(5-fluoro-3-oxo-spiro[4H-l,4-benzoxazin- 2,r-cyclobutene]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.020 g, 26.30%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.82 (s, 1H), 8.65 - 8.62 (m, 1H), 8.11 - 8.06 (m, 1H), 7.92 - 7.90 (m, 1H), 7.05 - 7.01 (m, 1H), 6.87 - 6.85 (m, 1H), 6.47 (s, 1H), 4.56 (s, 2H),
3.92 (s, 2H), 3.53 (s, 2H), 2.81 (d, j = 4.80 Hz, 3H), 2.53 - 2.47 (m, 2H), 2.27 - 2.21 (m, 2H), 1.94 - 1.89 (m, 1H), 1.81 - 1.74 (m, 1H).; LRMS (ES) m/z 457.3 (M++ 1).
Synthesis of example compound 50, 5-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyrazin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.030 g, 0.105 mmol), intermediate compound 7 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-pyrazin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.035 g, 0.110 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.057622 mL, 0.330 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 50 of 5-[2-[(2-ethyl-5-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]- 3,6-dihydrooxazin-5-yl]-N-methyl-pyrazin-2-carboxamide (0.010 g, 21.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.44 (s, 1H), 9.05 (d, J = 1.4 Hz, 1H), 8.93 (d, J = 1.4 Hz, 1H), 8.82 (d, J = 4.8 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.38 (t, J = 0.6 Hz, 1H), 7.18 (s, 1H), 4.73 (s, 2H), 4.12 (s, 2H), 3.66 (s, 2H), 2.85 - 2.80 (m, 5H), 1.22 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 425.2 (M++ 1).
Synthesis of example compound 51, 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyrazin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 0.048 g, 0.181 mmol), intermediate compound 7 of 5-(3,6-dihydro-2H-oxazin-5-yl)-N-methyl- pyrazin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N- diisopropylethylamine (100.00% solution, 0.09489 mL, 0.543 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 51 of 5-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6- dihydrooxazin-5-yl]-N-methyl-pyrazin-2-carboxamide (0.010 g, 10.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 8 11.85 (s, 1H), 9.06 (s, 1H), 8.97 (d, J = 1.4 Hz, 1H), 8.83 (d, J = 4.6 Hz, 1H), 8.43 (d, J = 1.8 Hz, 1H), 7.74 (d, J = 14.5 Hz, 2H), 7.21 (s, 1H), 4.77 (s, 2H), 4.11 (s, 2H), 3.68 (s, 2H), 2.84 (d, J = 4.0 Hz, 3H), 2.55 - 2.52 (m, 2H), 1.19 (t, J = 2.6 Hz, 3H).; LRMS (ES) m/z 407.4 (M++ 1).
Synthesis of example compound 52, 5-[2-[(2-ethyl-7-fhioro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-7-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 0.060 g, 0.200 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.073 g, 0.219 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.10911 mL, 0.625 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 52 of 5-[2-[(2-ethyl-7-fluoro- 3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin- 2-carboxamide (0.017 g, 20.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.66 (s, 1H), 8.66 (d, J = 4.9 Hz, 1H), 8.11 (q, J = 5.8 Hz, 1H), 7.93 (q, J = 3.2 Hz, 1H), 7.04 (d, J = 7.3 Hz, 1H), 6.88 (d, J = 10.4 Hz, 1H), 6.50 (s, 1H), 4.60 (s, 2H), 4.54 (t, J = 3.8 Hz, 1H), 3.89 (s, 2H), 3.56 (s, 2H), 2.79 (d, J = 4.8 Hz, 3H), 1.87 - 1.72 (m, 2H), 0.98 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 445.2 (M++ 1).
Synthesis of example compound 53, 5-[2-[(2-ethyl-8-fhioro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-8-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 0.060 g, 0.200 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.073 g, 0.219 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.10911 mL, 0.625 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with methanol and dried to obtain example compound 53 of 5-[2-[(2-ethyl-8-fluoro-3- oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide (0.010 g, 10.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.83 (s, 1H), 8.65 (t, J = 4.7 Hz, 1H), 8.11 (q, J = 5.8 Hz, 1H), 7.93 (q, J = 3.2 Hz, 1H), 6.87 (q, J = 4.4 Hz, 1H), 6.78 (s, 1H), 6.50 (s, 1H), 4.61 (q, J = 4.1 Hz, 3H), 3.85 (s, 2H), 3.53 (s, 2H), 2.79 (d, J = 4.8 Hz, 3H), 1.85 - 1.77 (m, 2H), 1.00 (t, J = 3.7 Hz, 3H).; LRMS (ES) m/z 456.2 (M+ + 1).
Synthesis of example compound 54, 6-fhioro-5-[2-[(5-fhioro-3-oxo-spiro[4H- l,4-benzoxazin-2,l'-cyclopentane]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl- pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-5-fluoro-spiro[4H-l,4-benzoxazin-2,r- cyclopentane]-3-one (100.00%, 0.050 g, 0.159 mmol), intermediate compound 2 of 5-(3,6- dihydro-2H-oxazin-5-yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 1.050 equiv., 0.167 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1386 mL, 0.796 mmol) were dissolved in acetonitrile (10 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 54 of 6-fluoro-5-[2-[(5-fluoro-3-oxo-spiro[4H-l,4-benzoxazin- 2,r-cyclopentane]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.046 g, 61.44%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.84 (s, 1H), 8.65 - 8.64 (m, 1H), 8.11 - 8.07 (m, 1H), 7.92 - 7.90 (m, 1H), 7.04 - 7.00 (m, 1H), 6.79 - 6.77 (m, 1H), 6.47 (s, 1H), 4.56 (s, 2H), 3.93 (s, 2H), 3.53 (s, 2H), 2.81 (d, j = 4.68 Hz, 3H), 2.07 - 2.04 (m, 2H), 1.88 - 1.69 (m, 6H).; LRMS (ES) m/z 471.2 (M++ 1).
Synthesis of example compound 55, 5-[2-[(5-fhioro-3-oxo-spiro[4H-l,4- benzoxazin-2,l'-cyclopentane]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin- 2-carboxamide A solution in which 6-(bromomethyl)-5-fluoro-spiro[4H-l,4-benzoxazin-2,l'- cyclopentane]-3-one (100.00%, 0.050 g, 0.159 mmol), intermediate compound 1 of 5-(3,6- dihydro-2H-oxazin-5-yl)-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 1.050 equiv., 0.167 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1386 mL, 0.796 mmol) were dissolved in acetonitrile (10 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 55 of 5-[2-[(5-fluoro-3-oxo-spiro[4H-l,4-benzoxazin-2,l'- cyclopentane]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.035 g, 48.61%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.84 (s, 1H), 8.71 - 8.70 (m, 1H), 8.64 (s, 1H), 7.96 (s, 2H), 7.04 - 7.00 (m, 1H), 6.79 - 6.77 (m, 1H), 6.57 (s, 1H), 4.66 (s, 2H), 3.93 (s, 2H), 3.51 (s, 2H), 2.81 (d, j = 4.72 Hz, 3H), 2.07 - 2.04 (m, 2H), 1.87 - 1.71 (m, 6H).; LRMS (ES) m/z 453.3 (M+ + 1).
Synthesis of example compound 56, N-ethyl-5-[2-[(2-ethyl-7-fluoro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-pyridin-2-carboxamide
A solution in which 2-ethyl-7-fluoro-3-oxo-4H-quinoxalin-6-carbaldehyde
(100.00%, 0.095 g, 0.429 mmol), intermediate compound 11 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-ethyl-6-fluoro-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.100 g, 0.300 mmol), and acetic acid (100.00% solution, 0.04935 mL, 0.859 mmol) were dissolved in dichloromethane (15 mL) was stirred at room temperature for 15 minutes and sodium triacetoxyborohydride (100.00%, 0.289 g, 1.362 mmol) was added and further stirred overnight at the same temperature. Solvent was removed from the reaction mixture under reduced pressure, and then water was poured into the resulting concentrate and extracted with dichloromethane. An organic layer was washed with a saturated sodium chloride aqueous solution, dehydrated with anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting concentrate was purified via column chromatography (SiCh, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated, and then the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 56 of N-ethyl-5-[2- [(2-ethyl-7-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro- pyridin-2-carboxamide (2.000 mg, 2.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.83 (s, 1H), 8.65 (t, J = 4.7 Hz, 1H), 8.11 (q, J = 5.8 Hz, 1H), 7.93 (q, J = 3.2 Hz, 1H), 6.87 (q, J = 4.4 Hz, 1H), 6.78 (s, 1H), 6.50 (s, 1H), 4.61 (q, J = 4.1 Hz, 3H), 3.85 (s, 2H), 3.53 (s, 2H), 2.79 (d, J = 4.8 Hz, 3H), 1.85 - 1.77 (m, 2H), 1.00 (t, J = 3.7 Hz, 3H).; LRMS (ES) m/z 456.2 (M+ + 1).
Synthesis of example compound 57, 4-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-2-fluoro-N-methyl-benzamide
A solution in which 6-(bromomethyl)-2-ethyl-5-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 50.000 mg, 0.174 mmol), intermediate compound 15 of 4-(3,6-dihydro-2H-oxazin- 5-yl)-2-fluoro-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (100.00%, 64.000 mg, 0.183 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.151 mL, 0.867 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 57 of 4-[2- [(2-ethyl-5-fluoro-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-2-fluoro- N-methyl-benzamide (56.600 mg, 73.55%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.83 (s, 1H), 8.18 - 8.17 (m, 1H), 7.61 - 7.59 (m, 1H), 7.32 - 7.29 (m, 2H), 7.03 - 7.01 (m, 1H), 6.82 - 6.80 (m, 1H), 6.50 - 6.49 (m, 1H), 4.58 - 4.57 (m, 2H), 4.54 - 4.53 (m, 1H), 3.91 (s, 2H), 3.48 - 3.47 (m, 2H), 2.76 (d, J = 4.6 Hz, 3H), 1.84 - 1.82 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 444.3 (M+ + 1).
Synthesis of example compound 58, 4-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-2-fhioro-N-methyl-benzamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 50.000 mg, 0.187 mmol), intermediate 15 of 4-(3,6-dihydro-2H-oxazin-5-yl)-2-fluoro-N- methyl-benzamide; 2,2,2-trifluoroacetic acid (100.00%, 69.000 mg, 0.197 mmol), and N,N- diisopropylethylamine (100.00% solution, 0.163 mL, 0.936 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then a precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 58 of 4-[2-[(7-ethyl-6-oxo- 5H-l,5-naphthyridin-3-yl)methyl]-3,6-dihydrooxazin-5-yl]-2-fluoro-N-methyl-benzamide (39.900 mg, 50.46%) in a white solid form. 'H-NMR (400 MHz, DMSO-d6) 5 11.85 (s, 1H), 8.43 - 8.42 (m, 1H), 8.19 - 8.18 (m, 1H), 7.75 - 7.74 (m, 1H), 7.68 - 7.67 (m, 1H), 7.62 - 7.60 (m, 1H), 7.34 - 7.31 (m, 2H), 6.56 - 6.55 (m, 1H), 4.61 - 4.60 (m, 2H), 4.06 - 4.05 (m, 2H), 3.57 - 3.56 (m, 2H), 2.77 (d, J = 4.6 Hz, 3H), 2.53 - 2.50 (m, 2H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 423.5 (M++ 1).
Synthesis of example compound 59, 4-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-2-fhioro-N-methyl-benzamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-pyrido[2,3-b][l,4]oxazine-2-one (100.00%, 50.000 mg, 0.184 mmol), intermediate 15 of 4-(3,6-dihydro-2H-oxazin-5-yl)-2- fluoro-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (100.00%, 68.000 mg, 0.194 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.161 mL, 0.924 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried. Then, methanol (10 mL) was added to the resulting product and stirred to filter out a precipitated solid, which was then washed with methanol and dried to obtain example compound 59 of 4-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3- b] [ 1 ,4] oxazin-7 -yl)methyl] -3 ,6-dihydrooxazin-5-yl] -2-fluoro-N-methyl-benzamide (60.600 mg, 77.06%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.77 (s, 1H), 8.18 - 8.17 (m, 1H), 7.76 (d, J = 2.0 Hz, 1H), 7.62 - 7.60 (m, 1H), 7.33 - 7.28 (m, 3H), 6.53 - 6.52 (m, 1H), 4.77 - 4.75 (m, 1H), 4.59 - 4.58 (m, 2H), 3.88 - 3.87 (m, 2H), 3.49 - 3.48 (m, 2H), 2.77 (d, J = 4.6 Hz, 3H), 1.90 -
1.87 (m, 2H), 1.01 (t, J = 7.6 Hz, 3H).; LRMS (ES) m/z 427.5 (M+ + 1). Synthesis of example compound 60, 6-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-5-fhioro-N-methyl-pyridin-3-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 35.000 mg, 0.131 mmol), intermediate 10 of 6-(3,6-dihydro-2H-oxazin-5-yl)-5-fluoro-N- methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 50.000 mg, 0.142 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.117 mL, 0.672 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 60 of 6-[2- [(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6-dihydrooxazin-5-yl]-5-fluoro-N- methyl-pyridin-3 -carboxamide (43.000 mg, 77.51%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 8 11.84 (s, 1H), 8.79 (s, 1H), 8.69 - 8.68 (m, 1H), 8.41 - 8.40 (m, 1H), 8.07 - 8.04 (m, 1H), 7.74 - 7.72 (m, 2H), 6.95 - 6.94 (m, 1H), 4.73 - 4.72 (m, 2H), 4.09 - 4.08 (m, 2H), 3.65 - 3.64 (m, 2H), 2.81 (d, J = 4.5 Hz, 3H), 2.53 - 2.50 (m, 2H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 424.5 (M+ + 1).
Synthesis of example compound 61, 6-[2-[(2-ethyl-5-fhioro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-5-fluoro-N-methyl-pyridin-3- carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-5-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 37.000 mg, 0.128 mmol), intermediate 10 of 6-(3,6-dihydro-2H-oxazin-5-yl)-5- fluoro-N-methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 49.000 mg, 0.140 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.115 mL, 0.660 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 61 of 6-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6- dihydrooxazin-5-yl]-5-fluoro-N-methyl-pyridin-3-carboxamide (48.500 mg, 84.98%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.82 (s, 1H), 8.78 - 8.77 (m, 1H), 8.68 - 8.67 (m, 1H), 8.05 - 8.03 (m, 1H), 7.05 - 7.03 (m, 1H), 6.90- 6.89 (m, 1H), 6.82 - 6.80 (m, 1H) 4.68 - 4.67 (m, 2H), 4.55 - 4.53 (m, 1H), 3.93 (s, 2H), 3.56 - 3.55 (m, 2H), 2.80 (d, J = 4.5 Hz, 3H), 1.84 - 1.81 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 445.5 (M+ + 1).
Synthesis of example compound 62, 6-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-5-fhioro-N-methyl-pyridin-3- carboxamide A solution in which 7-(bromomethyl)-3-ethyl-lH-pyrido[2,3-b][l,4]oxazine-2-one (100.00%, 35.000 mg, 0.129 mmol), intermediate 10 of 6-(3,6-dihydro-2H-oxazin-5-yl)-5- fluoro-N-methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 49.000 mg, 0.140 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.116 mL, 0.666 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 62 of 6-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7-yl)methyl]-3,6- dihydrooxazin-5-yl]-5-fluoro-N-methyl-pyridin-3-carboxamide (44.000 mg, 79.74%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.77 (s, 1H), 8.79 - 8.78 (m, 1H), 8.69 - 8.68 (m, 1H), 8.06 - 8.04 (m, 1H), 7.75 - 7.74 (m, 1H), 7.30 - 7.29 (m, 1H), 6.92 - 6.91 (m, 1H), 1.76 - 1.74 (m, 1H), 4.69 - 4.68 (m, 2H), 3.91 - 3.90 (m, 2H), 3.57 - 3.56 (m, 2H), 2.81 (d, J = 4.5 Hz, 3H), 1.89 - 1.84 (m, 2H), 1.00 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 428.6 (M++ 1).
Synthesis of example compound 63, 4-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-2-fluoro-N-methyl-benzamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 50.000 mg, 0.175 mmol), intermediate 15 of 4-(3,6-dihydro-2H-oxazin-5-yl)-2- fluoro-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (100.00%, 65.000 mg, 0.186 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.15 mL, 0.861 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then a precipitated solid was filtered out, washed with acetonitrile, and dried. Then, methanol (2 mL) and dichloromethane (2 mL) were added to the resulting product and stirred to filter out a precipitated solid, which was then washed with methanol and dried to obtain example compound 63 of 4-[2-[(2-ethyl-5-fluoro-3- oxo-4H-quinoxalin-6-yl)methyl] -3 ,6-dihydrooxazin-5-yl] -2-fluoro-N-methyl-benzamide (13.000 mg, 16.83%) in a white solid form.
XH-NMR (400 MHz, DMSO-d6) 6 12.40 (s, 1H), 8.17 - 8.16 (m, 1H), 7.59 - 7.55 (m, 2H), 7.35 - 7.33 (m, 3H), 6.53 - 6.52 (m, 1H), 4.59 - 4.58 (m, 2H), 4.07 - 4.06 (m, 2H), 3.56 - 3.55 (m, 2H), 2.82 (q, J = 7.4 Hz, 2H), 2.76 (d, J = 4.4 Hz, 3H), 1.22 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 441.5 (M+ + 1).
Synthesis of example compound 64, 6-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-3-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 35.000 mg, 0.131 mmol), intermediate 8 of 6-(3,6-dihydro-2H-oxazin-5-yl)-N-methyl-pyridin- 3-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 46.000 mg, 0.138 mmol), and N,N- diisopropylethylamine (100.00% solution, 0.114 mL, 0.655 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 64 of 6- [2- [(7 -ethyl-6-oxo-5H- 1 ,5 -naphthyridin-3 -yl)methyl] -3 ,6-dihydrooxazin-5-yl] -N -methyl- pyridin-3-carboxamide (38.400 mg, 72.29%) in a light yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 8 11.84 (s, 1H), 8.91 - 8.90 (m, 1H), 8.59 - 8.58 (m, 1H), 8.42 - 8.41 (m, 1H), 8.16 - 8.14 (m, 1H), 7.74 - 7.72 (m, 3H), 6.96 - 6.95 (m, 1H), 4.74 (s, 2H), 4.08 - 4.07 (m, 2H), 3.62 - 3.61 (m, 2H), 2.80 (d, J = 4.5 Hz, 3H), 2.52 (q, J = 7.4 Hz, 2H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 441.5 (M+ + 1).
Synthesis of example compound 65, 4-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-3-fhioro-N-methyl-benzamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 45.000 mg, 0.168 mmol), intermediate 16 of 4-(3,6-dihydro-2H-oxazin-5-yl)-3-fluoro-N- methyl-benzamide; 2,2,2-trifluoroacetic acid (100.00%, 62.000 mg, 0.177 mmol), and N,N- diisopropylethylamine (100.00% solution, 0.147 mL, 0.844 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 65 of 4-[2- [(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6-dihydrooxazin-5-yl]-3-fluoro-N- methyl-benzamide (52.200 mg, 73.36%) in a light yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 6 11.85 (s, 1H), 8.52 - 8.51 (m, 1H), 8.42 - 8.41 (m, 1H), 7.74 - 7.73 (m, 1H), 7.70 - 7.69 (m, 1H), 7.67 - 7.65 (m, 2H), 7.50 - 7.48 (m, 1H), 6.36 -
6.35 (m, 1H). 4.56 - 4.55 (m, 2H), 4.06 - 4.05 (m, 2H), 3.59 - 3.58 (m, 2H), 2.78 (d, J = 4.5 Hz, 3H), 2.53 (q, J = 7.4 Hz, 2H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 423.5 (M+ + 1). Synthesis of example compound 66, 6-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-3-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 36.000 mg, 0.126 mmol), intermediate 8 of 6-(3,6-dihydro-2H-oxazin-5-yl)-N- methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 44.000 mg, 0.132 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.11 mL, 0.632 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried. Then, the resulting product was purified via column chromatography (SiCL plate, 20x20x1 mm; methanol/methanol= 3%) and concentrated to obtain example compound 66 of 6-[2-[(2-ethyl-5-fluoro-3-oxo-4H-quinoxalin- 6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-3-carboxamide (7.000 mg, 13.09%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 8.90 - 8.89 (m, 1H), 8.61 - 8.60 (m, 1H), 8.15 - 8.13 (m, 1H), 7.70 - 7.68 (m, 1H), 7.54 - 7.52 (m, 1H), 7.37 - 7.35 (m, 1H), 6.94 - 6.93 (m, 1H), 4.70 - 4.69 (m, 2H), 4.10 - 4.09 (m, 2H), 3.60 - 3.59 (m, 2H), 2.80 - 2.79 (m, 5H), 1.23 - 1.21 (m, 3H).; LRMS (ES) m/z 424.5 (M+ + 1).
Synthesis of example compound 67, 4-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-3-fluoro-N-methyl-benzamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 45.000 mg, 0.158 mmol), intermediate compound 16 of 4-(3,6-dihydro-2H-oxazin- 5-yl)-3-fluoro-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (100.00%, 58.000 mg, 0.166 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.137 mL, 0.787 mmol) were dissolved in acetonitrile (2 mL) at room temperature was stirred overnight at 80°C, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 67 of 4-[2- [(2-ethyl-5-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-3-fluoro-N- methyl-benzamide (38.400 mg, 55.24%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 8.57 - 8.56 (m, 1H), 7.67 - 7.65 (m, 2H), 7.49 - 7.47 (m, 1H), 7.25 - 7.23 (m, 1H), 6.93 - 6.92 (m, 1H), 6.32 - 6.31 (m, 1H), 4.55 - 4.54 (m, 2H), 4.02 - 4.01 (m, 2H), 3.50 - 3.49 (m, 2H), 3.16 - 3.15 (m, 2H), 2.77 - 2.76 (m, 3H), 2.69 (q, J = 7.3 Hz, 1H), 0.85 - 0.83 (m, 3H).; LRMS (ES) m/z 441.0 (M++ 1).
Synthesis of example compound 68, N-ethyl-5-[2-[(2-ethyl-7-fluoro-3-oxo-4H- l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fhioro-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-7-fluoro-4H-l,4-benzoxazin-3-one
(100.00%, 0.045 g, 0.160 mmol), intermediate compound 11 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-ethyl-6-fluoro-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.057 g, 0.164 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.082 mL, 0.470 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 68 of N-ethyl-5-[2-[(2-ethyl- 7-fluoro-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-pyridin-2- carboxamide (1.500 mg, 2.10%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.67 (s, 1H), 8.70 (t, J = 6.0 Hz, 1H), 8.11 (t, J = 8.7 Hz, 1H), 7.93 (d, J = 7.8 Hz, 1H), 7.04 (d, J = 7.4 Hz, 1H), 6.88 (d, J = 10.4 Hz, 1H), 6.50 (s, 1H), 4.60 (s, 2H), 4.55 (t, J = 3.8 Hz, 1H), 3.89 (s, 2H), 3.56 (s, 2H), 3.38 (d, J = 7.1 Hz, 2H), 1.87 - 1.72 (m, 2H), 1.10 (t, J = 7.1 Hz, 3H), 0.98 (t, J = 7.3 Hz, 3H).; LRMS (ES) m/z 459.1 (M+ + 1).
Synthesis of example compound 69, N-ethyl-5-[2-[(2-ethyl-8-fluoro-3-oxo-4H- l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fhioro-pyridin-2-carboxamide
A solution in which 6-(bromomethyl)-2-ethyl-8-fluoro-4H-l,4-benzoxazin-3-one
(100.00%, 0.045 g, 0.160 mmol), intermediate compound 11 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N-ethyl-6-fluoro-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.057 g, 0.164 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.082 mL, 0.470 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 69 of N-ethyl-5-[2-[(2-ethyl- 8-fluoro-3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-pyridin-2- carboxamide (5.000 mg, 7.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.83 (s, 1H), 8.70 (t, J = 8.0 Hz, 1H), 8.11 - 8.09 (m, 1H), 7.93 (d, J = 7.6 Hz, 1H), 6.87 (d, J = 4.8 Hz, 1H), 6.79 (s, 1H), 6.50 (s, 1H), 4.62 - 4.59 (m, 3H), 3.85 (s, 2H), 3.53 (s, 2H), 3.26 (d, J = 10.8 Hz, 2H), 1.85 - 1.77 (m, 2H), 1.10 (t, J = 7.1 Hz, 3H), 1.00 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 459.1 (M+ + 1).
Synthesis of example compound 70, 6-fluoro-N-methyl-5-[2-[(2-methyl-3-oxo- 4H-pyrido[3,2-b][l,4]oxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin-2- carboxamide
A solution in which 6-(bromomethyl)-2-methyl-4H-pyrido[3,2-b][l,4]oxazin-3-one (100.00%, 0.081 g, 0.313 mmol), intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5- yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.100 g, 0.300 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.156 mL, 0.893 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 2%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 70 of 6-fluoro-N-methyl-5- [2- [(2-methyl-3 -oxo-4H-pyrido [3 ,2-b] [ 1 ,4] oxazin-6-yl)methyl] -3 ,6-dihydrooxazin-5- yl]pyridin-2-carboxamide (10.000 mg, 8.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 8 11.18 (s, 1H), 8.65 (d, J = 4.8 Hz, 1H), 8.11 (q, J = 5.8 Hz, 1H), 7.93 (q, J = 3.2 Hz, 1H), 7.36 (d, J = 8.1 Hz, 1H), 7.10 (d, J = 8.1 Hz, 1H), 6.50 (s, 1H), 4.74 (q, J = 2.7 Hz, 1H), 4.60 (s, 2H), 3.93 (s, 2H), 3.58 (s, 2H), 2.79 (d, J = 4.8 Hz, 3H), 1.44 (d, J = 6.8 Hz, 3H).; LRMS (ES) m/z 415.1 (M+ + 1).
Synthesis of example compound 71, 4-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-
3-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-benzamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-l,5-naphthyridin-2-one (100.00%, 0.053 g, 0.199 mmol), intermediate compound 3 of 4-(3,6-dihydro-2H-oxazin-5-yl)-N-methyl- benzamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N- diisopropylethylamine (100.00% solution, 0.099 mL, 0.567 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 71 of 4-[2-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]-3,6- dihydrooxazin-5-yl]-N-methyl-benzamide (20.000 mg, 30.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 11.85 (s, 1H), 8.43 - 8.41 (m, 2H), 7.81 (d, J = 8.3 Hz, 2H), 7.75 (s, 1H), 7.70 (s, 1H), 7.48 (d, J = 8.4 Hz, 2H), 6.44 (s, 1H), 4.46 (s, 2H), 4.07 (s, 2H), 3.58 (s, 2H), 2.77 (d, J = 4.5 Hz, 3H), 2.56 - 2.54 (m, 2H), 1.19 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 405.2 (M++ 1).
Synthesis of example compound 72, 4-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-benzamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.057 g, 0.199 mmol), intermediate compound 3 of 4-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N- diisopropylethylamine (100.00% solution, 0.099 mL, 0.567 mmol) were dissolved in acetonitrile (5 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 72 of 4-[2-[(2-ethyl-5-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]- 3,6-dihydrooxazin-5-yl]-N-methyl-benzamide (20.000 mg, 20.00%) in a yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.44 (s, 1H), 8.41 (d, J = 4.6 Hz, 1H), 7.80 (d, J = 8.5 Hz, 2H), 7.55 (d, J = 8.4 Hz, 1H), 7.47 (d, J = 8.5 Hz, 2H), 7.37 (t, J = 7.7 Hz, 1H), 6.47 (s, 1H), 4.62 (s, 2H), 4.08 (s, 2H), 3.56 (s, 2H), 2.83 - 2.77 (m, 5H), 1.22 (t, J = 7.2 Hz, 3H).; LRMS (ES) m/z 423.1 (M++ 1).
Synthesis of example compound 73, 4-[2-[(2-ethyl-5-fhioro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-benzamide
A solution in which 6-(bromomethyl)-2-ethyl-5-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 0.057 g, 0.199 mmol), intermediate compound 3 of 4-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N- diisopropylethylamine (100.00% solution, 0.099 mL, 0.567 mmol) were dissolved in acetonitrile (15 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 73 of 4-[2-[(2-ethyl-5-fluoro-3-oxo-4H-l,4-benzoxazin-6- yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-benzamide (20.000 mg, 20.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.84 (s, 1H), 8.41 (d, J = 4.6 Hz, 1H), 7.79 (d, J = 8.4 Hz, 2H), 7.46 (d, J = 8.4 Hz, 2H), 7.03 (t, J = 8.0 Hz, 1H), 6.81 (d, J = 8.4 Hz, 1H), 6.44 (s, 1H), 4.61 (s, 2H), 4.55 (q, J = 4.1 Hz, 1H), 3.92 (s, 2H), 34846.00 (s, 2H), 3.48 (s, 2H), 2.77 (d, J = 4.5 Hz, 3H), 1.85 - 1.74 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 426.2 (M+ + 1).
Synthesis of example compound 74, 4-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-benzamide
A solution in which 7-(bromomethyl)-3-ethyl-lH-pyrido[2,3-b][l,4]oxazin-2-one (100.00%, 0.054 g, 0.199 mmol), intermediate compound 3 of 4-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-benzamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N- diisopropylethylamine (100.00% solution, 0.099 mL, 0.567 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 74 of 4-[2-[(3-ethyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7- yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-benzamide (20.000 mg, 30.00%) in a white solid form. XH-NMR (400 MHz, DMSO-d6) 6 10.79 (s, 1H), 8.41 (d, J = 4.5 Hz, 1H), 7.81 - 7.76 (m, 3H), 7.47 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 2.0 Hz, 1H), 6.46 (s, 1H), 4.76 (q, J = 4.0 Hz, 1H), 4.61 (s, 2H), 3.89 (s, 2H), 3.50 (s, 2H), 2.77 (d, J = 4.5 Hz, 3H), 1.91 - 1.77 (m, 2H), 1.00 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 408.3 (M++ 1).
Synthesis of example compound 75, 6-[2-[(2-ethyl-7-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-5-fluoro-N-methyl-pyridin-3- carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-6-fluoro-lH-quinoxalin-2-one (100.00%, 0.051 g, 0.179 mmol), intermediate compound 10 of 6-(3,6-dihydro-2H-oxazin-5- yl)-5-fluoro-N-methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.099 mL, 0.567 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 75 of 6-[2-[(2-ethyl-7-fluoro- 3 -oxo-4H-quinoxalin-6-yl)methyl] -3 ,6-dihydrooxazin-5-yl] -5-fluoro-N-methyl-pyridin-3 - carboxamide (20.000 mg, 30.00%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.32 (s, 1H), 8.81 (d, J = 1.6 Hz, 1H), 8.70 (d, J = 4.5 Hz, 1H), 8.07 (q, J = 19.7 Hz, 1H), 7.55 - 7.53 (m, 2H), 6.98 (s, 1H), 4.82 (s, 2H), 4.09 (s, 2H), 3.70 (s, 2H), 2.82 - 2.81 (m, 5H), 1.22 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 442.6 (M+ + 1).
Synthesis of example compound 76, 6-[2-[(2-ethyl-7-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-3-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-6-fluoro-lH-quinoxalin-2-one (100.00%, 0.054 g, 0.189 mmol), intermediate compound 8 of 6-(3,6-dihydro-2H-oxazin-5- yl)-N-methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.09908 mL, 0.567 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 76 of 6-[2-[(2-ethyl-7-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]- 3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-3-carboxamide (20.000 mg, 20.00%) in a yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.31 (s, 1H), 8.92 (d, J = 1.8 Hz, 1H), 8.60 (d, J = 4.5 Hz, 1H), 8.16 (q, J = 3.5 Hz, 1H), 7.76 (d, J = 8.4 Hz, 1H), 7.55 - 7.53 (m, 2H), 6.98 (s, 1H), 4.83 (s, 2H), 4.09 (s, 2H), 3.66 (s, 2H), 2.82 - 2.80 (m, 5H), 1.22 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 424.4 (M++ 1).
Synthesis of example compound 77, 6-[2-[(2-ethyl-5-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-5-fluoro-N-methyl-pyridin-3- carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-8-fluoro-lH-quinoxalin-2-one (100.00%, 0.051 g, 0.179 mmol), intermediate compound 10 of 6-(3,6-dihydro-2H-oxazin-5- yl)-5-fluoro-N-methyl-pyridin-3-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.099 mL, 0.567 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated to obtain example compound 77 of 6-[2-[(2-ethyl-5-fluoro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-5-fluoro-N-methyl-pyridin-3-carboxamide (10.000 mg, 10.00%) in a yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.44 (s, 1H), 8.79 (t, J = 1.6 Hz, 1H), 8.68 (d, J = 4.6 Hz, 1H), 8.05 (q, J = 4.8 Hz, 1H), 7.56 (d, J = 8.2 Hz, 1H), 7.39 (t, J = 3.7 Hz, 1H), 6.93 (s, 1H), 4.70 (s, 2H), 4.11 (s, 2H), 3.64 (s, 2H), 2.83 - 2.80 (m, 5H), 1.22 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 442.5 (M++ 1).
Synthesis of example compound 78, 5-[2-[(2-ethyl-8-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide
A mixture in which 7-(bromomethyl)-3-ethyl-5-fluoro-lH-quinoxalin-2-one; hydrobromide (100.00%, 0.100 g, 0.273 mmol), intermediate compound 2 of 5-(3,6-dihydro- 2H-oxazin-5-yl)-6-fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.100 g, 0.285 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.25 mL, 1.431 mmol) were mixed in acetonitrile (3 mL) at room temperature was stirred at 80°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 78 of 5-[2-[(2-ethyl-8-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]-3,6- dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2-carboxamide (0.070 g, 55.70%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.43 (brs, 1H), 8.67 - 8.65 (m, 1H), 8.15 - 8.10 (m, 1H), 7.94 (dd, J = 7.7, 1.7 Hz, 1H), 7.18 (s, 1H), 7.12 (d, J = 11.3 Hz, 1H), 6.53 (s, 1H), 4.63 (s, 2H), 4.03 (s, 2H), 3.61 (s, 2H), 2.85 - 2.80 (m, 5H), 1.23 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 442.2 (M+ + 1).
Synthesis of example compound 79, 6-fhioro-5-[2-[(8-fhioro-3-oxo-spiro[4H- l,4-benzoxazin-2,l'-cyclopentane]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl- pyridin-2-carboxamide
A solution in which intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.050 g, 0.108 mmol), 6-(bromomethyl)-8-fluoro-spiro[4H-l,4-benzoxazin-2,r-cyclopentane]-3-one (100.00%, 1.050 equiv., 0.113 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.07487 mL, 0.430 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 79 of 6-fluoro-5-[2-[(8-fluoro-3-oxo-spiro[4H-l,4-benzoxazin- 2,r-cyclopentane]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.030 g, 59.34%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.84 (s, 1H), 8.66 - 8.64 (m, 1H), 8.13 - 8.08 (m, 1H), 7.93 - 7.91 (m, 1H), 6.88 - 6.85 (m, 1H), 6.79 (s, 1H), 6.50 (s, 1H), 4.59 (s, 2H), 3.86 (s, 2H), 3.32 (s, 2H), 2.79 (d, j = 4.80 Hz, 3H), 2.07 - 2.02 (m, 2H), 1.90 - 1.72 (m, 6H).; LRMS
(ES) m/z 471.2 (M+ + 1).
Synthesis of example compound 80: 6-fluoro-5-[2-[(8-fluoro-2-methyl-3-oxo-
4H-quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide; dihydrochloride
[Step 1] Synthesis of 6-fluoro-5-[2-[(8-fluoro-2-methyl-3-oxo-4H-quinoxalin-6- yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid
A solution in which intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.053 g, 0.114 mmol), 7-(bromomethyl)-5-fluoro-3-methyl-lH-quinoxalin-2-one; hydrobromide (100.00%, 1.050 equiv., 0.120 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.07936 mL, 0.456 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. The reaction mixture was purified via column chromatography (Waters, C18; 0.1%- trifluoroacetic acid aqueous solution/acetonitrile = 0 to 80%) and concentrated to obtain 6- fluoro-5- [2- [(8 -fluoro-2-methyl-3 -oxo-4H-quinoxalin-6-yl)methyl] -3 ,6-dihydrooxazin-5-yl] - N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (0.018 g, 29.19%) in a white solid form.
[Step 2] Synthesis of example compound 80
To a solution in which 6-fluoro-5-[2-[(8-fluoro-2-methyl-3-oxo-4H-quinoxalin-6- yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.010 g, 0.018 mmol) were dissolved in dichloromethane (2 mL)/methanol (2 mL) at room temperature, hydrogen chloride (ca. 1 mol/L in ethyl acetate, 1.00 M solution, 0.03694 mL, 0.037 mmol) was added and stirred at the same temperature for one hour. Solvent was removed from the reaction mixture under reduced pressure, and then a precipitated solid was filtered out, washed with diethyl ether, and dried to obtain example compound 80 of 6- fluoro-5- [2- [(8 -fluoro-2-methyl-3 -oxo-4H-quinoxalin-6-yl)methyl] -3 ,6-dihydrooxazin-5-yl] - N-methyl-pyridin-2-carboxamide; dihydrochloride (0.008 g, 86.57%) in a yellow solid form.
'H-NMR (400 MHz, DMSO-d6) 6 12.44 (s, 1H), 8.66 - 8.65 (m, 1H), 8.14 - 8.09 (m, 1H), 7.94 - 7.92 (m, 1H), 7.16 (s, 1H), 7.12 - 7.10 (m, 1H), 6.52 (s, 1H), 4.62 (s, 2H), 4.02 (s, 2H), 3.60 (s, 2H), 2.80 (d, j = 4.80 Hz, 3H), 2.41 (s, 3H).; LRMS (ES) m/z 428.2 (M++ 1).
Synthesis of example compound 81, 5-[2-[(2-ethyl-7-fhioro-3-oxo-4H- quinoxalin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N,4-dimethyl-pyridin-2-carboxamide
A solution in which 7-(bromomethyl)-3-ethyl-6-fluoro-lH-quinoxalin-2-one (100.00%, 0.054 g, 0.189 mmol), intermediate compound 9 of 5-(3,6-dihydro-2H-oxazin-5- yl)-N,4-dimethyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.060 g, 0.200 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.09908 mL, 0.567 mmol) were dissolved in acetonitrile (20 mL) at 80°C was stirred overnight at the same temperature, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 12 g cartridge; methanol/dichloromethane = 0 to 3%) and concentrated. Then, the resulting product was crystallized with methanol (10 mL) at 70°C and filtered to obtain a solid, which was then washed with diethyl ether and dried to obtain example compound 81 of 5-[2-[(2-ethyl-7-fluoro-3-oxo-4H-quinoxalin-6-yl)methyl]- 3,6-dihydrooxazin-5-yl]-N,4-dimethyl-pyridin-2-carboxamide (10.000 mg, 10.00%) in a white solid form. 'H-NMR (400 MHz, DMSO-d6) 6 12.35 (s, 1H), 8.73 (d, J = 4.9 Hz, 1H), 8.35 (s, 1H), 7.91 (s, 1H), 7.55 (d, J = 10.5 Hz, 1H), 7.50 (d, J = 7.0 Hz, 1H), 6.01 (s, 1H), 4.48 (s, 2H), 4.09 (s, 2H), 3.61 (s, 2H), 2.81 - 2.80 (m, 5H), 2.41 (s, 3H), 1.21 (t, J = 7.4 Hz, 3H).; LRMS (ES) m/z 438.4 (M+ + 1).
Synthesis of example compound 82, 6-fhioro-5-[2-[(7-fhioro-3-oxo-spiro[4Hl,4-benzoxazin-2,l'-cyclopentane]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl- pyridin-2-carboxamide
A solution in which intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.080 g, 0.172 mmol), 6-(bromomethyl)-7-fluoro-spiro[4H-l,4-benzoxazin-2,r-cyclopentane]-3-one (100.00%, 1.050 equiv., 0.181 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1198 mL, 0.688 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 82 of 6-fluoro-5-[2-[(7-fluoro-3-oxo-spiro[4H-l,4-benzoxazin-2,l'- cyclopentane]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.058 g, 71.70%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.66 (s, 1H), 8.67 - 8.64 (m, 1H), 8.13 - 8.08/ (m, 1H), 7.94 - 7.91 (m, 1H), 7.06 - 7.04 (m, 1H), 6.85 - 6.83 (m, 1H), 6.50 (s, 1H), 4.61 (s, 2H), 3.89 (s, 2H), 3.55 (s, 2H), 2.79 (d, j = 4.80 Hz, 3H), 2.07 - 2.02 (m, 2H), 1.87 - 1.71 (m, 6H).; LRMS (ES) m/z 471.2 (M++ 1). Synthesis of example compound 83, 6-fhioro-5-[2-[(5-fhioro-3-oxo-spiro[4Hl,4-benzoxazin-2,l'-cyclohexane]-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl- pyridin-2-carboxamide
A solution in which intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.080 g, 0.172 mmol), 6-(bromomethyl)-5-fluoro-spiro[4H-l,4-benzoxazin-2,r-cyclohexane]-3-one
(100.00%, 1.050 equiv., 0.181 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1198 mL, 0.688 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; ethyl acetate/hexane = 0 to 100%) and concentrated to obtain example compound 83 of 6-fluoro-5- [2- [(5-fluoro-3 -oxo- spiro [4H- 1 ,4-benzoxazin-2, 1 '-cyclohexane] -6-yl)methyl] -3,6- dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.065 g, 78.03%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.75 (s, 1H), 8.65 - 8.62 (m, 1H), 8.11 - 8.06 (m, 1H), 7.92 - 7.90 (m, 1H), 7.05 - 7.01 (m, 1H), 6.86 - 6.84 (m, 1H), 6.47 (s, 1H), 4.57 (s, 2H), 3.92 (s, 2H), 3.52 (s, 2H), 2.79 (d, j = 4.76 Hz, 3H), 1.75 - 1.56 (m, 9H), 1.26 - 1.22 (m, 1H). ; LRMS (ES) m/z 485.2 (M++ 1).
Synthesis of example compound 84, 6-fluoro-N-methyl-5-[2-[(3-methyl-2-oxo- lH-pyrido[3,4-b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]pyridin-2- carboxamide
A solution in which intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.080 g, 0.172 mmol), 7-(bromomethyl)-3-methyl-lH-pyrido[3,4-b][l,4]oxazin-2-one; hydrobromide (100.00%, 1.050 equiv., 0.181 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1198 mL, 0.688 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; methanol/dichloromethane = 0 to 20%) and concentrated to obtain example compound 84 of 6-fluoro-N-methyl-5- [2- [(3 -methyl-2-oxo- 1 H-pyrido [3 ,4-b] [ 1 ,4]oxazin-7 -yl)methyl] -3,6- dihydrooxazin-5-yl]pyridin-2-carboxamide (0.013 g, 18.29%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.99 (s, 1H), 8.66 - 8.65 (m, 1H), 8.12 - 8.08 (m, 2H), 7.95 - 7.93 (m, 1H), 7.10 (s, 1H), 6.52 (s, 1H), 4.80 - 4.78 (m, 1H), 4.63 (s, 2H), 3.95 (s, 2H), 3.59 (s, 2H), 2.79 (d, j = 4.76 Hz, 3H), 1.45 (d, j = 6.80 Hz, 3H).; LRMS (ES) m/z 414.2 (M++ 1).
Synthesis of example compound 85, 5-[2-[(2,2-diethyl-5-fluoro-3-oxo-4H-l,4- benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2- carboxamide
A solution in which intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.080 g, 0.172 mmol), 6-(bromomethyl)-2,2-diethyl-5-fluoro-4H-l,4-benzoxazin-3-one (100.00%, 1.050 equiv., 0.181 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1198 mL, 0.688 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCL, 4 g cartridge; ethyl acetate/hexane = 0 to 100%) and concentrated to obtain example compound 85 of 5-[2-[(2,2-diethyl-5-fluoro- 3-oxo-4H-l,4-benzoxazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin- 2-carboxamide (0.070 g, 86.17%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.80 (s, 1H), 8.65 - 8.63 (m, 1H), 8.11 - 8.06 (m, 1H), 7.92 - 7.90 (m, 1H), 7.02 - 6.98 (m, 1H), 6.79 - 6.77 (m, 1H), 6.47 (s, 1H), 4.56 (s, 2H), 3.91 (s, 2H), 3.52 (s, 2H), 2.79 (d, j = 4.80 Hz, 3H), 1.88 - 1.82 (m, 2H), 1.71 - 1.66 (m, 2H), 0.90 - 0.87 (m, 6H).; LRMS (ES) m/z 473.1 (M+ + 1).
Synthesis of example compound 86, 5-[2-[(3-ethyl-2-oxo-lH-pyrido[3,4- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fhioro-N-methyl-pyridin-2- carboxamide
A solution in which intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.080 g, 0.172 mmol), 7-(bromomethyl)-3-ethyl-lH-pyrido[3,4-b][l,4]oxazin-2-one (100.00%, 1.050 equiv., 0.181 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1198 mL, 0.688 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. Solvent was removed from the reaction mixture under reduced pressure, and then the resulting concentrate was purified via column chromatography (SiCh, 4 g cartridge; methanol/dichloromethane = 0 to 20%) and concentrated to obtain example compound 86 of 5-[2-[(3-ethyl-2-oxo-lH-pyrido[3,4-b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-6- fluoro-N-methyl-pyridin-2-carboxamide (0.013 g, 18.29%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 11.06 - 10.98 (m, 1H), 8.66 - 8.65 (m, 1H), 8.14 - 8.09 (m, 2H), 7.95 - 7.92 (m, 1H), 7.08 (s, 1H), 6.52 (s, 1H), 4.65 - 4.62 (m, 3H), 3.94 (s, 2H), 3.59 (s, 2H), 2.79 (d, j = 4.84 Hz, 3H), 1.88 - 1.76 (m, 2H), 1.01 - 0.97 (m, 3H).; LRMS (ES) m/z 428.2 (M+ + 1).
Synthesis of example compound 87, 6-fhioro-5-[2-[(5-fhioro-2-methyl-3-oxo-
4H-l,4-benzothiazin-6-yl)methyl]-3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2- carboxamide
A solution in which intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.100 g, 0.215 mmol), 6-(bromomethyl)-5-fluoro-2-methyl-4H-l,4-benzothiazin-3-one (100.00%, 1.050 equiv., 0.226 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1497 mL, 0.860 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 87 of 6-fluoro-5-[2-[(5-fluoro-2-methyl-3-oxo-4H-l,4-benzothiazin-6-yl)methyl]- 3,6-dihydrooxazin-5-yl]-N-methyl-pyridin-2-carboxamide (0.063 g, 65.65%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.54 (s, 1H), 8.66 - 8.62 (m, 1H), 8.11 - 8.07 (m, 1H), 7.92 - 7.90 (m, 1H), 7.17 - 7.09 (m, 2H), 6.49 (s, 1H), 4.56 (s, 2H), 3.96 (s, 2H), 3.72 - 3.70 (m, 1H), 3.56 (s, 2H), 2.79 (d, j = 4.80 Hz, 3H), 1.33 - 1.32 (m, 3H).; LRMS (ES) m/z 447.1 (M++ 1).
Synthesis of example compound 88, 5-[2-[(3,3-dimethyl-2-oxo-lH-pyrido[2,3- b][l,4]oxazin-7-yl)methyl]-3,6-dihydrooxazin-5-yl]-6-fhioro-N-methyl-pyridin-2- carboxamide A solution in which intermediate compound 2 of 5-(3,6-dihydro-2H-oxazin-5-yl)-6- fluoro-N-methyl-pyridin-2-carboxamide; 2,2,2-trifluoroacetic acid (100.00%, 0.080 g, 0.172 mmol), 7-(bromomethyl)-3,3-dimethyl-lH-pyrido[2,3-b][l,4]oxazin-2-one (100.00%, 1.010 equiv., 0.174 mmol), and N,N-diisopropylethylamine (100.00% solution, 0.1198 mL, 0.688 mmol) were dissolved in acetonitrile (5 mL) at room temperature was stirred at 70°C for 18 hours, after which the temperature was lowered to room temperature to terminate the reaction. A precipitated solid was filtered out, washed with acetonitrile, and dried to obtain example compound 88 of 5-[2-[(3,3-dimethyl-2-oxo-lH-pyrido[2,3-b][l,4]oxazin-7-yl)methyl]-3,6- dihydrooxazin-5-yl]-6-fluoro-N-methyl-pyridin-2-carboxamide (0.038 g, 51.71%) in a white solid form.
'H-NMR (400 MHz, DMSO-d6) 6 10.74 (s, 1H), 8.65 - 8.64 (m, 1H), 8.12 - 8.07 (m, 1H), 7.93 - 7.91 (m, 1H), 7.91 (s, 1H), 6.29 (s, 1H), 6.49 (s, 1H), 4.57 (s, 2H), 3.90 (s, 2H), 3.54 (s, 2H), 2.79 (d, j = 4.76 Hz, 3H), 1.44 (s, 6H).; LRMS (ES) m/z 428.1 (M++ 1).
< Experimental Example >
Experimental Example 1. Evaluation of PARP1/PARP2 binding ability (in vitro) 1. Experimental method
The PARP1/PARP2 binding ability of the test substance was measured using PARPtrap™ Assay Kit for PARP1 (BPS Bio #80584)/PARP2 (BPS Bio #78296). For a PARP1 assay, samples were treated at a concentration of 0.1 nM, 0.3 nM, 1 nM, 10 nM, 100 nM, 1,000 nM, and 10,000 nM. For a PARP2 assay, samples were treated at a concentration of 10 nM, 30 nM, 100 nM, 300 nM, 1,000 nM, 3,000 nM, and 10,000 nM. After sample treatment, the reaction was allowed to proceed for 60 minutes at room temperature, and fluorescence polarization (Ex 485 nm/Em 535 nm) was measured. 2. Experimental results
The PARP1/PARP2 binding ability EC50 (nM) obtained according to the above experimental method is shown in a following table.
[Table 1]
From the above results, it can be confirmed that the compounds according to examples of the present invention have very good binding ability to PARP1.
In other words, it can be confirmed that the compounds according to examples of the present invention exhibit an excellent inhibitory effect against PARP1 and an excellent selective inhibitory effect against PARP1.
While the present invention have been described in detail above, it is apparent to those skilled in the art that such detailed descriptions are set forth to illustrate exemplary embodiments only, but are not construed to limit the scope of the present invention. Thus, it should be understood that the substantial scope of the present invention is defined by the accompanying claims and equivalents thereto.

Claims

1. A compound represented by following formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof:
[Formula 1] wherein,
M is CRX, N, NRy, O, or S,
X, Y and Z are each independently N or CRZ, each of Rx, Ry and Rz is independently H, halogen, C1-C5 alkyl, or C1-C5 haloalkyl,
RA and RB are each independently H, halogen, OH, CN, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, or O-(C1-C5 alkyl), or RA and RB are bonded to each other to form cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl,
“““ is a single bond or a double bond, is absent or a single bond,
Ri and R2 are each independently H, halogen, OH, CN, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, or O-(C1-C5 alkyl),
A is cycloalkylene, cycloalkenylene, heterocycloalkylene, heterocycloalkenylene, arylene, or heteroarylene, and halogen is F, Cl, Br or I.
2. The compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof of claim 1, wherein in above formula 1,
M is CRX, N, NRy, O, or S,
X, Y and Z are each independently N or CRZ, each of Rx, Ry and Rz is independently H, halogen, C1-C5 alkyl, or C1-C5 haloalkyl, RA and RB are each independently H, halogen, OH, CN, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, or O-(C1-C5 alkyl), or RA and RB are bonded to each other to form three- to seven-membered cycloalkyl, three- to seven-membered cycloalkenyl, three- to seven-membered heterocycloalkyl including one to three heteroatoms independently selected from the group consisting of N, O and S, or three- to seven-membered heterocycloalkenyl including one to three heteroatoms independently selected from the group consisting of N, O and S, is a single bond or a double bond, is absent or a single bond,
Ri and R2 are each independently H, halogen, OH, CN, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, or O-(C1-C5 alkyl),
A is 3- to 7-membered cycloalkylene, three- to seven-membered cycloalkenylene, three- to seven-membered heterocycloalkylene including one to three heteroatoms independently selected from the group consisting of N, O and S, three- to seven-membered heterocycloalkenylene including one to three heteroatoms independently selected from the group consisting of N, O and S, C6-C12 arylene, or five- to 12-membered heteroarylene including one to three heteroatoms independently selected from the group consisting of N, O and S, and halogen is F, Cl, Br or I.
3. The compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof of claim 1, wherein in above formula 1,
M is CRX, N, O or S,
X, Y and Z are each independently N or CRZ, each of Rx and Rz is independently H or halogen,
RA and RB are each independently H or C1-C5 alkyl, or RA and RB are bonded to each other to form three- to seven-membered cycloalkyl, is a single bond or a double bond, is absent or a single bond,
Ri and R2 are each independently H, halogen, C1-C5 alkyl, C1-C5 haloalkyl, or O- (C1-C5 alkyl),
A is C6-C12 arylene or five- to 12-membered heteroarylene including one to three heteroatoms independently selected from the group consisting of N, O and S, and halogen is F.
4. The compound represented by formula 1, stereoisomers thereof, or pharmaceutically acceptable salts thereof of claim 1, wherein in above formula 1,
M is CRX, N, O or S,
X, Y and Z are each independently N or CRZ, each of Rx and Rz is independently H or halogen,
RA and RB are each independently H or C1-C3 alkyl, or RA and RB are bonded to each other to form four- to six-membered cycloalkyl, is a single bond or a double bond, is absent or a single bond, Ri and R2 are each independently H, halogen, C1-C3 alkyl, C1-C3 haloalkyl, or O- (C1-C3 alkyl),
A is phenylene or five- to six-membered heteroarylene including one to three Ns, and halogen is F.
5. A compound described in a following table, stereoisomers thereof, or pharmaceutically acceptable salts thereof:
6. A pharmaceutical composition comprising the compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof according to any one of claims 1 to 5 as an active ingredient for preventing or treating PARP1 activity-associated diseases.
7. The pharmaceutical composition of claim 6, wherein the PARP1 activity-associated diseases are cancer or inflammatory diseases.
8. The pharmaceutical composition of claim 7, wherein the cancer is at least one selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, hematological cancer, gastrointestinal cancer, lung cancer, and brain cancer.
9. The pharmaceutical composition according to claim 7, wherein the inflammatory disease is at least one selected from the group consisting of rheumatoid arthritis, multiple sclerosis, Crohn's disease, ulcerative colonitis, ulcerative colitis, graft-versus-host disease (GVHD), systemic lupus erythematosus, toxic shock syndrome, osteoarthritis, and insulindependent diabetes mellitus.
10. A method for preventing or treating PARP1 activity-associated diseases including administering a therapeutically effective amount of the compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof according to any one of claims 1 to 5 or the pharmaceutical composition according to any one of claims 6 to 9.
11. A use of the compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof according to any one of claims 1 to 5 or the pharmaceutical composition according to any one of claims 6 to 9 for preventing or treating PARP1 activity-associated diseases.
12. A use of the compound, stereoisomers thereof, or pharmaceutically acceptable salts thereof according to any one of claims 1 to 5 or the pharmaceutical composition according to any one of claims 6 to 9 in preparing a medicament for preventing or treating PARP1 activity-associated diseases.
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US11795158B2 (en) 2020-06-25 2023-10-24 Astrazeneca Ab Chemical compounds

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WO2010111626A2 (en) * 2009-03-27 2010-09-30 Takeda Pharmaceutical Company Limited Poly (adp-ribose) polymerase (parp) inhibitors
CN116891456A (en) * 2022-04-08 2023-10-17 上海翰森生物医药科技有限公司 Heterocyclic derivative inhibitor, preparation method and application thereof
WO2023227052A1 (en) * 2022-05-25 2023-11-30 西藏海思科制药有限公司 Bicyclic derivative parp inhibitor and use thereof
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