CN116478145B - ALK2 kinase inhibitors - Google Patents

ALK2 kinase inhibitors Download PDF

Info

Publication number
CN116478145B
CN116478145B CN202310387713.7A CN202310387713A CN116478145B CN 116478145 B CN116478145 B CN 116478145B CN 202310387713 A CN202310387713 A CN 202310387713A CN 116478145 B CN116478145 B CN 116478145B
Authority
CN
China
Prior art keywords
compound
mmol
group
deuterated
stereoisomer
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.)
Active
Application number
CN202310387713.7A
Other languages
Chinese (zh)
Other versions
CN116478145A (en
Inventor
杨欣
崔荣
温俏冬
殷建明
郑鹛
吕裕斌
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.)
Hangzhou Bangshun Pharmaceutical Co ltd
Original Assignee
Hangzhou Bangshun Pharmaceutical Co ltd
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 Hangzhou Bangshun Pharmaceutical Co ltd filed Critical Hangzhou Bangshun Pharmaceutical Co ltd
Publication of CN116478145A publication Critical patent/CN116478145A/en
Application granted granted Critical
Publication of CN116478145B publication Critical patent/CN116478145B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4433—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with oxygen as a ring hetero atom
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with 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
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • 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
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/06—Antianaemics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/81—Amides; Imides
    • C07D213/82—Amides; Imides in position 3
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • 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
    • 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
    • C07D471/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
    • C07D491/20—Spiro-condensed systems
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

The invention discloses an ALK2 kinase inhibitor shown in a general formula I, a pharmaceutical composition, a preparation method and application thereof in preparing, preventing and/or treating drugs related to ALK2 signal channels. The invention is characterized in thatThe compounds are ideal high activity ALK2 kinase inhibitors and are useful in the treatment and/or prevention of diseases including anemia, inflammation, neoplasms, and partially rare diseases, such as progressive fibrodysplasia ossificans, diffuse endogenous bridge glioma, iron refractory iron deficiency anemia, inflammatory anemia, myelodysplastic syndrome, multiple myeloma, and in the treatment of MPN-related anemia in combination with JAK2 inhibitors.

Description

ALK2 kinase inhibitors
Technical Field
The invention belongs to the field of pharmaceutical chemistry, and relates to an ALK2 kinase inhibitor, a pharmaceutical composition, a preparation method and application thereof in preparing, preventing and/or treating drugs related to ALK2 signal channel indications.
Background
ALK2 (also known as the ACVR1 protein), an activin receptor-like kinase, a Bone Morphogenic Protein (BMP) type i receptor, is one of the members of the TGF- β family of transforming growth factors. Binding of the ligand to the type II receptor induces phosphorylation of the type I receptor, which affects cell function by recruiting and phosphorylating downstream receptor-associated Smad and further regulating expression of the associated gene (PLoS One 2013Apr 30;8 (4): e 62721).
Abnormal activation of ALK2 was found to lead to the development of a variety of diseases. For example, a significant proportion of patients with progressive ossified Fibrodysplasia (FOP) find mutations in ALK 2R 206H, which enhance downstream SMAD signaling and overactivate BMP pathways. ACVR1 mutations are present in 20-32% of diffuse endogenous bridge gliomas (DIPG). ALK2 has also been found to play a role in cancer of the ovaries, prostate, and erythroleukemia (Cells 2019,8,1366).
Furthermore, in the study of Laura Silvesti et al (Vitam Horm.2019; 110:71-99), ALK2 was found to be a key receptor regulating the up-regulation of iron-dependent hepcidin expression, whereas high levels of hepcidin resulted in iron-restricted erythropoiesis, manifested as Iron Refractory Iron Deficiency Anaemia (IRIDA) and inflammatory Anaemia (AI). Inflammatory anemia, also known as chronic Anemia (ACD), is a disease of anemia that is globally secondary to iron deficiency anemia, and is common in chronic or hospitalized patients. Inflammatory anemia refers to anemia resulting from chronic inflammation caused by chronic infections (including tuberculosis, acquired immunodeficiency syndrome), autoimmune diseases (such as rheumatoid arthritis, lupus erythematosus, inflammatory bowel disease, etc.), and certain tumors (including hematological tumors associated with increased cytokines such as hodgkin's lymphoma, myelofibrosis, MDS, MM, etc., common solid tumors such as ovarian cancer, lung cancer, etc.). Such inflammatory anaemia can severely affect patient quality of life, physiological function and reduce survival (N Engl J Med 2019; 381:1148-57). In the studies of Maria Vittoria Verga Falzacappa et al (Blood 2007;109 (1): 353-358), IL-6 was then revealed to be the major cytokine associated with inflammatory anaemia, which upregulates the expression of hepcidin via the JAK2-STAT3 pathway. Patent WO2021102258 discloses the use of ALK2 inhibitors in combination with JAK2 inhibitors for the treatment of myeloproliferative neoplasms (MPNs) and related anemias. Suggesting the potential for application of ALK2 inhibitors in this indication.
Disclosure of Invention
The invention aims to provide an ALK2 kinase inhibitor, a pharmaceutical composition, a preparation method and application thereof in preparing, preventing and/or treating drugs related to ALK2 signal channel indications.
In order to solve the defects in the prior art, the invention discloses a compound with a structure shown in a general formula (I), deuterated matters, stereoisomers or pharmaceutically acceptable salts thereof:
wherein,
ring A is a 5-10 membered heteroaryl, optionally substituted with 1-3R 3 Substitution;
ring B is a 6-15 membered cyclic group, and optionally 1-3 ring carbon atoms are replaced by oxygen; ring B is optionally substituted with 1 to 3R 4 Substitution;
L 1 selected from chemical bonds, C 1-6 Alkyl, imino, carbonyl, -CO-NH-, phenyl, 5-6 membered heteroaryl, -phenyl-CO-NH-;
L 2 selected from chemical bonds, - (CR) a R b ) n -, a part of imino group ether linkage, -C 1-6 alkyl-O-;
R 1 selected from C 1-6 Alkyl, 5-10 membered cycloalkyl, 5-10 membered heterocycloalkyl, and optionally substituted with 1-3R 5 Substitution;
R 2 selected from H, halogen, hydroxy, cyano, amino, or
R 2 Selected from 4-10 membered heterocycloalkyl or cycloalkyl, and optionally substituted with 1-3R 6 Substitution;
R 3 selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 An alkyl group;
R 4 selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, halo C 1-6 An alkyl group;
R 5 selected from halogen, hydroxy, nitro, cyano, amino, -CO-NR c R d 、C 1-6 Alkyl, C 1-6 Alkoxy, - (CR) a R b ) n -OH；
R 6 Selected from halogen, hydroxy, cyano, -CO-R c 、-CO-NR c R d 、-SO 2 -R c Or (b)
R 6 Selected from C 1-6 Alkyl, C 1-6 Alkoxy, 4-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, and optionally substituted with 1-3R e Substitution;
R a 、R b independently selected from H, halogen, hydroxy, nitro, cyano, amino or C 1-6 Alkyl, or
R a 、R b Form a 3-6 membered cycloalkyl group with the attached carbon atom;
R c 、R d independently selected from H, halogen, hydroxy, nitro, cyano, amino or C 1-6 An alkyl group;
R e selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 An alkoxy group;
n is an integer from 1 to 3.
In some preferred embodiments:
ring A is a 5-10 membered heteroaryl, optionally substituted with 1-3R 3 Substitution;
ring B is a 6-15 membered cyclic group, and optionally 1-3 ring carbon atoms are replaced by oxygen; ring B is optionally substituted with 1 to 3R 4 Substitution;
L 1 selected from chemical bonds, C 1-6 Alkyl, imino, carbonyl, -CO-NH-, phenyl, 5-6 membered heteroaryl, -phenyl-CO-NH-;
L 2 selected from chemical bonds, - (CR) a R b ) n -, a part of imino group ether linkage, -C 1-6 alkyl-O-;
R 1 selected from C 1-6 Alkyl, 5-10 membered cycloalkyl, 5-10 membered heterocycloalkyl, and optionally substituted with 1-3R 5 Substitution;
R 2 selected from 4-10 membered heterocycloalkyl, R 2 Optionally by 1-3R 6 Substitution;
R 3 selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 An alkyl group;
R 4 selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 Alkyl, halogenated C 1-6 An alkyl group;
R 5 selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 Alkyl radicals(CR a R b ) n -OH；
R 6 Selected from hydroxy, -CO-R c 、-CO-NR c R d 、-SO 2 -R c Or (b)
R 6 Selected from C 1-6 Alkyl, 4-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, and optionally substituted with 1-3R e Substitution;
R a 、R b independently selected from H, halogen, hydroxy, nitro, cyano, amino or C 1-6 Alkyl, or
R a 、R b Form a 3-6 membered cycloalkyl group with the attached carbon atom;
R c 、R d independently selected from H, halogen, hydroxy, nitro, cyano, amino or C 1-6 An alkyl group;
R e selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 An alkoxy group;
n is an integer from 1 to 3.
In some more preferred embodiments:
ring A is a 6-10 membered heteroaryl group containing 1-3 heteroatoms selected from N, O, S, optionally substituted with 1-3R 3 Substitution;
ring B is selected from 6-9 membered carbocyclyl or 6-13 membered heterocyclyl, and optionally 1 ring carbon atom is replaced by oxygen; ring B is optionally substituted with 1 to 3R 4 Substitution;
L 1 selected from imino, carbonyl, -CO-NH-, 5-6 membered heteroaryl, -phenyl-CO-NH-;
L 2 selected from chemical bonds, - (CR) a R b ) n -、-C 1-3 alkyl-O-;
R 1 selected from C 1-3 Alkyl, 6-8 membered cycloalkyl, 6-8 membered heterocycloalkyl, and optionally substituted with 1-3R 5 Substitution;
R 2 selected from 4-9 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, S, R 2 Optionally by 1-3R 6 Substitution;
R 3 selected from cyano, amino, C 1-3 An alkyl group;
R 4 selected from halogen, C 1-3 Alkyl, halogenated C 1-3 An alkyl group;
R 5 selected from halogen, hydroxy, cyano, C 1-6 Alkyl, - (CR) a R b ) n -OH；
R 6 Selected from hydroxy, -CO-R c 、-CO-NR c R d 、-SO 2 -R c Or (b)
R 6 Selected from C 1-6 Alkyl, 4-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, and optionally substituted with 1-3R e Substitution;
R a 、R b independently selected from H or C 1-3 Alkyl, or
R a 、R b Form a 3-6 membered cycloalkyl group with the attached carbon atom;
R c 、R d independently selected from H or C 1-3 An alkyl group;
R e selected from halogen, cyano, C 1-3 An alkoxy group;
n is an integer from 1 to 3.
In some preferred embodiments:
ring A is selected from
As preferable:
L 1 selected from imino, -CO-NH-, carbonyl,-phenyl-CO-NH-;
R 1 selected from the group consisting ofC 1-3 Alkyl, said R 1 Optionally by 1-3R 5 Substitution;
R 3 selected from cyano, amino, C 1-3 An alkyl group;
R 5 selected from halogen, cyanoHydroxy, C 1-6 Alkyl, - (CR) a R b ) n -OH；
R a 、R b Independently selected from H or C 1-3 An alkyl group;
n is an integer from 1 to 3.
In some preferred embodiments:
ring B is selected from
In some more preferred embodiments:
ring B is selected from
In some preferred embodiments:
L 2 selected from chemical bonds, - (CR) a R b ) n -、-C 1-3 alkyl-O-;
R 2 selected from, H, halogen, hydroxy, cyano, or
R 2 Selected from the group consisting of And optionally is substituted with 1-3R 6 Substitution;
R 4 selected from halogen, C 1-3 Alkyl, halogenated C 1-3 An alkyl group;
R 6 selected from halogen, hydroxy, -CO-R c 、-CO-NR c R d 、-SO 2 -R c Or (b)
R 6 Selected from C 1-6 Alkyl group,C 1-6 Alkoxy group,Optionally by 1-3R e Substitution; r is R a 、R b Independently selected from H or C 1-3 Alkyl, or
R a 、R b Form a 3-6 membered cycloalkyl group with the attached carbon atom;
R c 、R d independently selected from H or C 1-3 An alkyl group;
R e selected from halogen, hydroxy, cyano, C 1-3 An alkoxy group;
n is an integer from 1 to 3.
In certain embodiments according to the present invention, the compounds provided herein further have a structure represented by formula (II):
X 1 、X 2 independently selected from CH or N;
R 1 selected from 5-10 membered cycloalkyl, 5-10 membered heterocycloalkyl, optionally substituted with 1-3R 5 Substitution;
R 5 selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 Alkyl, - (CR) a R b ) n -OH；
Ring B isY 1 、Y 2 、Y 3 Independently selected from CH or N, wherein ring C is a 5-6 membered ring group in parallel, and optionally 1 ring carbon atom in ring C is replaced by oxygen; ring B is optionally substituted with 1 to 3R 4 Substitution;
R 4 selected from halogen, C 1-3 Alkyl, halogenated C 1-3 An alkyl group;
R 2 is a 4-8 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S and optionally containing 1-3R 6 Substitution;
R 6 selected from the group consisting of-CO-R c 、-SO 2 -R c Or (b)
R 6 Selected from C 1-6 Alkyl, 4-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, optionally substituted with 1-3R e Substitution;
R a 、R b independently selected from H, halogen, hydroxy, nitro, cyano, amino or C 1-6 An alkyl group;
R c independently selected from H, hydroxy, amino or C 1-6 An alkyl group;
R e selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 An alkoxy group;
n is an integer from 1 to 3.
In some preferred embodiments:
R 1 selected from the group consisting of
R 5 Selected from hydroxy, halogen, C 1-6 Alkyl, - (CR) a R b ) n -OH；
R a 、R b Independently selected from H, C 1-3 An alkyl group;
n is an integer from 1 to 3.
In some more preferred embodiments:
R 1 selected from the group consisting of
R 5 Selected from hydroxy, halogen, C 1-3 Alkyl, -CH 2 OH、-C(CH 3 ) 2 OH。
In certain embodiments according to the present invention, the compounds provided herein further have a structure represented by formula (II-1):
ring B isY 1 、Y 2 、Y 3 Independently selected from CH or N, wherein ring C is a 5-6 membered ring group in parallel, and optionally 1 ring carbon atom in ring C is replaced by oxygen; ring B is optionally substituted with 1 to 3R 4 Substitution;
R 4 selected from halogen, C 1-3 Alkyl, halogenated C 1-3 An alkyl group;
R 2 is a 5-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O, S and optionally containing 1-3R 6 Substitution;
R 6 selected from the group consisting of-CO-R c 、-SO 2 -R c Or (b)
R 6 Selected from C 1-6 Alkyl, 4-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, optionally substituted with 1-3R e Substitution;
R c independently selected from H, hydroxy, amino or C 1-6 An alkyl group;
R e selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 An alkoxy group.
In some preferred embodiments:
ring B is selected from
In some preferred embodiments:
ring B isRing C is a 5 membered carbocyclyl or heterocyclyl group in parallel, optionally 1 ring carbon atom in ring C being replaced by oxygen; ring B is optionally substituted with 1 to 3R 4 Substitution;
R 4 selected from halogen, C 1-3 Alkyl, halogenated C 1-3 An alkyl group.
In some more preferred embodiments:
ring B is selected from
Ring B is optionally substituted with 1 to 3R 4 Substitution;
R 4 selected from halogen, C 1-3 Alkyl, halogenated C 1-3 An alkyl group.
In some preferred embodiments:
R 2 selected from the group consisting ofPreferably And optionally is substituted with 1R 6 Substitution;
R 6 selected from the group consisting of-CO-R c 、-SO 2 -R c Or (b)
R 6 Selected from C 1-3 Alkyl, 4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl, optionally substituted with 1-3R e Substituted;
R c independently selected from H or C 1-3 An alkyl group;
R e selected from halogen, cyano, C 1-3 An alkoxy group.
In some more preferred embodiments:
R 2 selected from the group consisting ofPreferably->And is covered with 1R 6 Substitution;
R 6 is a 4-6 membered heterocycloalkyl group containing 1O atom, preferablyMore preferably
In certain embodiments according to the present invention, the compounds provided herein further have a structure represented by formula (III):
ring B isRing C is a parallel 5-6 membered cyclic group optionally containing 1-2 heteroatoms selected from N; ring B is optionally substituted with 1-2R 4 Substitution;
R 4 Selected from halogen, C 1-3 Alkyl, halogenated C 1-3 An alkyl group;
R 1 selected from 4-8 membered cycloalkyl, 4-8 membered heterocycloalkyl containing 2 heteroatoms selected from N, O, S, preferably More preferablyR 1 Optionally by 1-3R 5 Substitution;
R 5 selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 Alkyl, - (CR) a R b ) n -OH, preferably hydroxy, halogen, C 1-3 Alkyl, -CH 2 OH、-C(CH 3 ) 2 OH；
L 2 Selected from chemical bonds, - (CR) a R b ) n -；
R 2 Heterocyclylalkyl containing 1-2 heteroatoms selected from N, O for 5-6 members, optionally substituted with 1-2R 6 Substitution;
R 6 selected from halogen, hydroxy, amino, cyano, or
R 6 Selected from 4-7 membered cycloalkyl, 4-7 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, C 1-3 Alkyl, and optionally substituted with 1-3R e Substitution;
R a 、R b independently selected from H or C 1-3 Alkyl, or R a 、R b Form a 3-6 membered cycloalkyl group with the attached carbon atom;
R e is halogen;
n is an integer of 1 to 3.
In some preferred embodiments:
ring B is selected from
In some more preferred embodiments:
ring B is selected from
L 2 Selected from chemical bonds, C 1-3 An alkylene group;
R 2 heterocyclylalkyl containing 1-2 heteroatoms selected from N, O for 5-6 members, optionally substituted with 1-2R 6 Substitution;
R 6 selected from halogen, or
R 6 Selected from 4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, C 1-3 Alkyl, and optionally substituted with 1-3R e And (3) substitution.
In some preferred embodiments:
ring B is selected from
R 1 Selected from the group consisting ofOptionally by 1R 5 Substitution;
R 5 selected from hydroxy, -CH 2 OH；
L 2 Selected from the group consisting of chemical bonds;
R 2 heterocyclylalkyl containing 1-2 heteroatoms selected from N, O for 5-6 members, optionally substituted with 1R 6 Substitution;
R 6 selected from 4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl containing 1O, C 1-3 Alkyl, and optionally substituted with 1-3R e Substitution;
R e is halogen.
In some preferred embodiments:
R 2 selected from the group consisting ofPreferably->
R 6 Selected from the group consisting ofCyclohexyl, C 1-3 Alkyl, preferablyC 1-3 An alkyl group.
In some more preferred embodiments:
R 1 and R is R 5 The following structure is formed:in some preferred embodiments:
ring B is selected from
R 1 Selected from the group consisting ofOptionally by 1R 5 Substitution;
R 5 selected from hydroxy, -CH 2 OH；
L 2 Selected from the group consisting of chemical bonds;
R 2 heterocyclylalkyl containing 1-2N for 5-6 membered ring, substituted with 1R 6 Substitution;
R 6 selected from 4-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O.
In some preferred embodiments:
R 2 selected from the group consisting ofPreferably->R 6 Selected from the group consisting ofPreferably->In some more preferred embodiments:
R 1 and R is R 5 The following structure is formed:in some preferred embodiments:
ring B is selected from
R 1 Selected from the group consisting ofOptionally by 1R 5 Substitution;
R 5 selected from hydroxy, -CH 2 OH；
L 2 Selected from chemical bonds, C 1-3 An alkylene group;
R 2 heterocyclylalkyl containing 1-2 heteroatoms selected from N, O for 5-6 members, optionally substituted with 1-2R 6 Substitution;
R 6 selected from halogen, or
R 6 Selected from 4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl containing 1O, and optionally substituted with 1-3R e Substitution;
R e is halogen.
In some preferred embodiments:
R 2 selected from the group consisting ofPreferably
R 6 Selected from halogen, or
R 6 Selected from the group consisting ofCyclohexyl, preferablyAnd optionally is substituted with 1-3R e Substitution;
R e is halogen.
In some more preferred embodiments:
R 1 and R is R 5 The following structure is formed:
in certain embodiments according to the present invention, the compounds provided herein further have a structure represented by formula (IV):
X 1 、X 2 independently selected from CH or N;
R 1 selected from 4-8 membered cycloalkyl, 4-8 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, S, preferably More preferablyOptionally by 1-2R 5 Substitution;
R 5 selected from C 1-3 Alkyl, C 1-3 Alkoxy, -C 1-3 alkyl-OH, halogen, OH, amino, amido, cyano;
ring B isr, p and q are integers of 1-3; z is Z 1 、Z 2 、Z 3 Independently selected from CH or N, and up to one is N; />Is a single bond or a double bond; when->Z is a single bond 4 Is CH 2 Or O, Z 5 Is CH 2 Or NH; when->In the case of double bonds, Z 4 Is CH, Z 5 CH or N; z is Z 6 Is NH or O; optionally 1 ring carbon atom in ring B is replaced by oxygen; ring B is optionally substituted with 1-2R 4 Substitution;
R 4 selected from halogen, C 1-3 Alkyl, C 1-3 Alkoxy, hydroxy;
L 2 Is a chemical bond, - (CR) a R b ) n -；
R 2 Selected from H, halogen, hydroxy, cyano, amino, or
R 2 Selected from 4-6 membered heterocycloalkyl or 4-6 membered cycloalkyl, preferably 4-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, S or 4-6 membered cycloalkyl, more preferablyFurther preferably-> And optionally is substituted with 1-2R 6 Substitution;
R 6 selected from halogen, hydroxy, and CC 1-3 Alkyl, C 1-3 Alkoxy, cyano;
R a 、R b independently selected from H or C 1-3 Alkyl, or R a 、R b Form a 3-6 membered cycloalkyl group with the attached carbon atom;
n is an integer of 1 to 3.
In some preferred embodiments:
ring B is selected from
In some preferred embodiments:
ring B isr or q is an integer from 1 to 3; z is Z 1 、Z 2 Or Z is 3 Independently selected from CH or N, and up to one is N; />Is a single bond or a double bond; when->Z is a single bond 4 Is CH 2 Or O; when->Z is a double bond 4 CH; ring B is optionally substituted with 1-2R 4 Substitution;
R 4 selected from halogen, C 1-3 Alkyl, C 1-3 Alkoxy, hydroxy.
In some more preferred embodiments:
X 1 、X 2 CH;
ring B isr is an integer from 1 to 2, < >>Is a single bond or a double bond.
In some more preferred embodiments:
ring B isPreferably->* Terminal to pyridine, terminal to L 2 Connection, r is an integer from 1 to 2, ">Is a single bond or a double bond.
L 2 Is a chemical bond or C 1-3 An alkylene group;
R 2 Selected from H, halogen, hydroxy, cyano, amino, or
R 2 Selected from 4-6 membered heterocycloalkyl or 4-6 membered cycloalkyl, preferably 4-6 membered contains 1-2 members selected from N, O,Heterocycloalkyl of S heteroatom or 4-6 membered cycloalkyl, more preferablyFurther preferred is And optionally is substituted with 1-2R 6 Substitution;
R 6 selected from halogen, hydroxy, C 1-3 Alkyl, C 1-3 Alkoxy, cyano.
In some preferred embodiments:
R 1 selected from the group consisting ofPreferablyOptionally by 1-2R 5 Substitution;
R 5 selected from halogen, OH, C 1-3 Alkyl, C 1-3 Alkoxy, -C 1-3 alkyl-OH, preferably F, cl, -OH, methyl, -CH 2 -OH；
Preferably R 1 And R is R 5 The following structure is formed:
in some preferred embodiments:
ring B is selected from Preferably Further preferably->* Terminal to pyridine, terminal to L 2 Connecting;
L 2 is a chemical bond, C 1-3 Alkylene, preferably a bond or isopropyl;
R 2 is H orPreferably H or->
In certain embodiments according to the present invention, the compounds provided herein further have the structure of formula (V):
m is selected from CH or N;
R 6a selected from C 1-3 Alkoxy, preferably methoxy, ethoxy, propoxy;
R 6b selected from C 1-6 Alkyl, 4-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, preferably methyl, ethyl, isopropyl,Isopropyl is more preferred.
The compound represented by the general formula (1) includes the following specific compounds:
/>
/>
/>
/>
/>
/>
/>
In another aspect, the present invention also provides a pharmaceutical composition comprising the above compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier therefor.
In another aspect, the invention also provides application of the compound, deuterated compound, stereoisomer or pharmaceutically acceptable salt thereof or the pharmaceutical composition in preparing medicines for treating and/or preventing ALK 2-related diseases.
Preferably, the ALK 2-related diseases include anemia, inflammation, neoplasm, and a portion of ALK 2-related genetic diseases.
Preferably, the ALK 2-associated disorders include progressive fibrodysplasia ossificans, diffuse endogenous bridge glioma, iron refractory iron deficiency anemia, inflammatory anemia, myelodysplastic syndrome, multiple myeloma, myeloproliferative tumor-associated anemia.
Preferably, the medicament is used alone or in combination with other therapeutic agents.
Preferably, the other therapeutic agent is a JAK2 inhibitor for the treatment of myeloproliferative neoplasm-related anemia.
The compound provided by the invention has strong drug effect, good drug substitution property and low toxic and side effects, and is an ideal ALK2 inhibitor. The compound provided by the invention has better ALK2 inhibition activity, and the ALK2 inhibition activity is far greater than ALK5, so that the compound has better selectivity. The compounds of the present invention are ideal high activity ALK2 inhibitors and are useful in the treatment and/or prevention of diseases including anemia, inflammation, tumors, and partially rare diseases such as progressive fibrodysplasia ossificans, diffuse endogenous bridge glioma, iron refractory iron deficiency anemia, inflammatory anemia, myelodysplastic syndrome, multiple myeloma, and anemia associated with myeloproliferative tumors treated with JAK2 inhibitors.
Definition of the definition
As used herein, "optional" or "optionally" means that any selectable item may be selected or not. For example, "group a is optionally substituted with 1 to 3 groups B" includes four cases where group a is not substituted with group B, group a is substituted with 1 group B, group a is substituted with 2 groups B, and group a is substituted with 3 groups B.
"substituted" or "substituted" as used herein refers to any one or more hydrogen atoms on any atom of a group or fragment being substituted with a substituent, and may include heavy hydrogen and hydrogen variants, provided that the valence of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =o), meaning that two hydrogen atoms are replaced with oxygen, when the oxygen-substituted atom is a carbon atom, a carbonyl group is formed (c=o). Oxygen substitution does not occur on the aromatic group. The term "optionally substituted" means that the substituents may or may not be substituted, and the types and numbers of substituents may be arbitrary on the basis of being chemically realizable unless otherwise indicated.
When any variable (e.g., R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0 to 2R, the group may optionally be substituted with up to two R's, and R's in each case have independent options. Furthermore, combinations of substituents and/or variants thereof are only permissible if such combinations result in stable compounds.
When the number of one linking group is 0, such as- (CR) a R b ) 0 -it is meant that the linking group is a single bond/chemical bond.
When one of the variables is selected from the group consisting of a bond/single bond, the two groups representing the connection are directly connected, e.g. A-L 1 -R 1 Middle L 1 Representing a single bond means that the structure is in fact A-R 1 。
When a substituent is absent, it is meant that the substituent is absent, e.g., X in A-X is absent, meaning that the structure is actually A. When the listed substituents do not indicate which atom is attached to the substituted group, such substituents may be bonded through any atom thereof, for example, a pyridyl group may be attached to the substituted group as a substituent through any carbon atom on the pyridine ring.
When the exemplified linking group does not indicate its linking direction, its linking direction is arbitrary, e.g., rings A-L 1 -R 1 Medium linking group L 1 is-M-W-, in this case-M-W-can either connect the rings A and R in the same direction as the reading sequence from left to right 1 To form the ring A-M-W-R 1 The rings A and R may be connected in a direction opposite to the reading order from left to right 1 To form the ring A-W-M-R 1 . Combinations of such linking groups, substituents and/or variants thereof are permissible only if such combinations result in stable compounds.
Where a group has one or more bondable sites, any one or more of the sites of the group may be bonded to other groups by chemical bonds, unless specifically indicated. When the chemical bond is not positionedWhen H atoms exist in the connectable site, the number of H atoms in the site is correspondingly reduced along with the number of the connected chemical bonds to become a corresponding valence group. The chemical bond of the site and other groups can be straight with straight solid line bondDotted bond->And (3) representing. For example-OCH 3 The straight solid line bond in (a) represents the connection to other groups through the oxygen atom in the group; />The straight dashed bonds in (a) represent the attachment to other groups through both ends of a carbon atom in the group; />The dashed lines in (a) represent the attachment to other groups through carbon atoms in the 1 and 4 positions of the phenyl group; />It means that any of the ligatable sites on the piperidinyl group may be linked to other groups by 1 chemical bond, including at least +.>These 4 connection forms, even though the H atom is depicted on-N-, are +.>Still include->The group of this linkage is only when 1 chemical bond is linked, the H at this site will be correspondingly reduced by 1 to the corresponding monovalent piperidinyl group.
In spiro or fused ring groups, if not specified, a group or fragmentThe ligation site is located on the loop that is connected by the dotted line. For example, the number of the cells to be processed,any of the attachment sites on the nitrogen-containing spiro ring representing this group may be attached to other groups via 1 chemical bond, including at least +.>These 3 connection modes; />Any attachment site on the benzene ring representing the group may be attached to other groups by 1 chemical bond, including at leastThese 4 connection modes.
Unless otherwise specified, in a bridged ring group, the attachment site of the group or fragment may be any attachment site. For example, the number of the cells to be processed,any of the attachment sites on the nitrogen-containing spiro ring representing this group may be attached to other groups via 1 chemical bond, including at least +.>These 2 connection modes.
In general, the double bond and the single bond in the aromatic ring or the heteroaromatic ring are not limited. For example, the number of the cells to be processed,all refer to benzene rings or phenyl groups, in which the double bond or +.>In particular to a large pi bond of delocalization on the benzene ring plane; />In which, when ring C is defined as a 5-membered aryl or heteroaryl group, ring B and ring C constitute a conjugated and cyclic aryl or a cyclic heteroaryl group, in which case non-limiting examples thereof include-> Even->The left-hand ring of the middle contains only two double bonds, but +. >Still include->Such groups, in which case the double bond represents a large pi bond delocalized over the fused aromatic ring.
As used herein, numerical intervals include endpoints and any numerical values between the endpoints. For example, "0-3" may include 0, 1, 2, or 3, and "1-3" may include 1, 2, or 3.
"C" as used herein 1-n "include C 1-2 、C 1-3 、……C 1-n . For example, "C 1-6 "group" means having 1 to 6 carbon atoms in the moiety, i.e., the group contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms. Thus, for example, "C 1-4 Alkyl "means an alkyl group containing 1 to 4 carbon atoms, i.e. the alkyl group is selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. Numerical ranges, such as "1-6" herein refer to individual integers in the given range.
Ring member atoms refer to non-hydrogen atoms in the ring group that are used to form a ring. For exampleThe ring members of (a) being 3 carbon atomsA seed; />The middle ring atoms are three carbon atoms and 1 oxygen atom; />The intermediate ring atoms are 1N atom and 5 carbon atoms;the ring segment atoms are 8 carbon atoms and 1 nitrogen atom.
As used herein, "n-m-membered" refers to the number of ring segment atoms in the ring radical. For example, a "3-8 membered" group means that there are 3-8 ring atoms in the moiety, i.e., the group contains 3 ring atoms, 4 ring atoms, 5 ring atoms, 6 ring atoms, 7 ring atoms, or 8 ring atoms. Thus, for example, by "3-8 membered cycloalkyl" is meant a saturated cyclic hydrocarbon radical containing 3-8 carbon atoms, i.e. the alkyl radical is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl.
The term "hydrocarbyl" as used herein, alone or in combination, refers to an atomic group consisting of only carbon and hydrogen elements, including saturated, unsaturated, or aromatic hydrocarbon groups, such as alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl. Unless specifically stated, the "hydrocarbyl" may be straight-chain, branched, or cyclic.
The term "alkyl" as used herein, alone or in combination, refers to an optionally substituted straight chain or optionally substituted branched saturated aliphatic hydrocarbon. The "alkyl" group herein may preferably have 1 to 6 carbon atoms, for example, 1 to 5 carbon atoms, or 1 to 4 carbon atoms, or 1 to 3 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, 2-methyl-l-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-l-butyl, 2-methyl-3-butyl, 2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-l-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2-methyl2-dimethyl-l-butyl, 3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, hexyl and the like. In the radicals defined herein, where the number range appears as "alkyl", for example, "C 1-6 Alkyl "refers to an alkyl group that may be made up of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms, and alkyl groups herein also include those where no numerical range is specified. The alkyl group may be optionally substituted or unsubstituted.
"alkyl" as used herein in combination refers to an alkyl group attached to other groups, e.g., an alkyl group in an alkoxy group, as defined above when used alone.
The term "alkylene" as used herein, alone or in combination, refers to a saturated aliphatic divalent hydrocarbon group resulting from the removal of two hydrogen atoms from a straight or branched saturated aliphatic hydrocarbon group. The "alkylene" herein may preferably have 1 to 6 carbon atoms, for example 1 to 5 carbon atoms, or 1 to 4 carbon atoms, or 1 to 3 carbon atoms. Non-limiting examples of alkylene groups include-CH 2 - (i.e. methylene), -CH 2 -CH 2 - (i.e. ethylene), -CH 2 -CH 2 -CH 2 -、-CH(CH 3 )CH 2 -、-C(CH 3 ) 2 -、-CH 2 -C(CH 3 )-CH 2 -、-CH 2 -CH 2 -CH 2 -CH 2 -、-CH 2 -C(CH 3 )-CH 2 -CH 2 -、-CH 2 -CH 2 -CH 2 -CH 2 -CH 2 -、-CH 2 -CH 2 -CH 2 -CH 2 -CH 2 -CH 2 -and the like. The alkylene group may be optionally substituted or unsubstituted.
The term "alkenyl" as used herein, alone or in combination, refers to an optionally substituted straight chain or optionally substituted unsaturated aliphatic hydrocarbon having one or more carbon-carbon double bonds. The "alkenyl" herein may preferably have 1 to 6 carbon atoms, for example 1 to 5 carbon atoms, or 1 to 4 carbon atoms, or 1 to 3 carbon atoms. Non-limiting examples of alkyl groups include ethenyl, propenyl, 1-butenyl, 2-butenyl, 1-pentenyl, 1-hexenyl, 1, 3-butadienyl, 1, 3-pentadienyl, 1, 4-pentadienyl and 1, 4-hexadienyl groups/moieties. Unless otherwise indicated, the term "alkenyl" does not include "cycloalkenyl".
The term "alkoxy" or "-O-alkyl" as used herein, alone or in combination, means "alkyl-O-". Non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy and the like. Alkoxy groups may be optionally substituted or unsubstituted.
The term "cyclic group" or "ring" as used herein, alone or in combination, refers to any organic compound having a cyclic structure, wherein the cyclic group may be saturated or unsaturated (including aryl) and may include one or more heteroatoms such as N, O or S in its carbon skeleton. Examples of cyclic groups include carbocyclyl and heterocyclyl groups as discussed below, and in particular may be cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl and heteroaryl. When the cyclic group is bicyclic or polycyclic, any of the rings is optionally selected from cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, or heteroaryl; when one or more of the rings is an aryl group, the remaining rings may be aryl groups, or cycloalkyl, heterocycloalkyl, cycloalkenyl, or heterocycloalkenyl groups having no aromaticity. The number of rings in the cyclic group may be monocyclic, bicyclic or polycyclic. Generally, bicyclic or polycyclic ring groups may be classified as spiro, bridged (including fused or fused) rings, depending on the manner of attachment. Unless otherwise specified, a cyclic group is a 3-15 membered cyclic group, meaning that it contains 3 to 12 ring members, preferably a 3-12 membered cyclic group, more preferably a 3-10 membered cyclic group.
In the definition of "cyclic group", the term "spiro" refers to a cyclic group having two or more cyclic structures and single rings sharing one atom (called spiro atom) with each other. Preferably 7-15 membered, more preferably 8-9 membered. The spiro ring is classified into a single spiro group, a double spiro group or a multiple spiro group according to the number of common spiro atoms between rings, and herein, preferably a single spiro group and a double spiro group, preferably a 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered spiro group. Non-limiting examples thereof include, but are not limited to, spiro [3.5] nonane, bicyclo [3.2.1] octane.
In the definition of "cyclic group," bridged ring "refers to a polycyclic cyclic group containing two or more cyclic structures and sharing two or more atoms. Wherein "fused ring" or "condensed ring" is a special bridged ring, and refers to a polycyclic ring group having two or more cyclic structures and sharing a pair of atoms with each other. Preferably 7-15 membered, more preferably 8-9 membered. The number of constituent rings may be classified as a bicyclic, tricyclic, tetracyclic or polycyclic bridged ring group, preferably a bicyclic or tricyclic ring, and herein preferably a bicyclic or tricyclic ring, more preferably a 5-membered/5-membered, 5-membered/6-membered or 6-membered/6-membered bicyclic bridged ring group. Non-limiting examples thereof include, but are not limited to, bicyclo [2.2.2] octane, spiro [3.4] octane.
Unless otherwise indicated, spiro and bridged rings may also be used in cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl groups discussed below, including spirocycloalkyl, spirocycloalkenyl, spiroheterocycloalkenyl, bridged cycloalkyl, bridged cycloalkenyl, bridged heterocycloalkyl, and bridged heterocycloalkenyl groups. Condensed rings include condensed ring alkyl, condensed ring alkenyl, condensed ring aryl, and condensed ring heteroaryl. The above terms are defined similarly to spiro, bridged, parallel or fused rings.
The term "carbocyclyl" as used herein, alone or in combination, includes cycloaliphatic and aryl groups, including monocyclic, fused, bridged and spiro rings. Examples of heterocyclyl groups include cycloalkyl, cycloalkenyl, and aryl groups as discussed below. Preferred herein are 3-10 membered monocyclic, bicyclic or tricyclic carbocyclyl groups.
The term "cycloalkyl" as used herein, alone or in combination, refers to a saturated monocyclic, bicyclic or polycyclic carbocycle, which may be a spiro or bridged ring. Preferred herein are 3-12 membered cycloalkyl groups, more preferred are 3-10 membered cycloalkyl groups, and most preferred are 3-8 membered cycloalkyl groups. Non-limiting examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like cycloalkyl groups, which may be optionally substituted or unsubstituted.
In the definition of "cycloalkyl", a "spiro" refers to an all-carbon polycyclic group having two or more cyclic structures and single rings sharing one carbon atom (referred to as a spiro atom) with each other. Preferably 6 to 12 membered, more preferably 8 to 9 membered. The spirocyclic groups are classified into single-, double-or multi-spirocyclic groups according to the number of common spiro atoms between rings, and herein, single-and double-spirocyclic groups are preferable, and 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered spirocyclic groups are preferable.
In the definition of "cycloalkyl", a "fused ring" refers to an all-carbon polycyclic group containing two or more cyclic structures and having a pair of carbon atoms common to each other. Preferably 6 to 12 membered, more preferably 8 to 9 membered. The number of constituent rings may be classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyl group, and is herein preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered, 5-membered/6-membered or 6-membered/6-membered bicycloalkyl group.
In the definition of "cycloalkyl" a "bridged ring" refers to an all-carbon polycyclic group containing two or more cyclic structures and sharing two carbon atoms not directly attached to each other. Preferably 6 to 12 membered, more preferably 8 to 9 membered. Cycloalkyl groups which may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged according to the number of constituent rings, preferably bicyclic or tricyclic.
The term "cycloalkenyl" as used herein, alone or in combination, refers to a monocyclic, bicyclic or polycyclic, non-aromatic, carbocycle having 1 or more carbon-carbon double bonds, which may be spiro or bridged. Preferred herein are 3-12 membered cycloalkenyl groups, more preferred are 3-10 membered cycloalkenyl groups, and most preferred are 3-8 membered cycloalkenyl groups. Non-limiting examples of cycloalkenyl groups include, but are not limited to, cyclopent-1-en-1-yl, cyclohex-1-en-1-yl, and cyclohex-1, 3-dien-1-yl, which may be optionally substituted or unsubstituted.
The term "aryl" as used herein, alone or in combination, refers to an aromatic hydrocarbyl ring. The term "aryl" includes mono-cyclic aromatic hydrocarbons and polycyclic fused ring aromatic hydrocarbons in which all fused ring systems (excluding any ring systems that are part of or formed by optional substituents) are aromatic. Examples of aryl groups/moieties include phenyl, naphthyl, anthryl and phenanthryl. Unless otherwise indicated, the term "aryl" does not include "heteroaryl".
The term "heterocyclyl" as used herein, alone or in combination, includes alicyclic and heteroaryl groups wherein one or more (such as one, two, three or four) ring members are heteroatoms, such as oxygen, nitrogen, sulfur atoms, and the like, including monocyclic, fused, bridged and spiro rings. Examples of heterocyclyl groups include heterocycloalkyl, heterocycloalkenyl, and heteroaryl groups as discussed below. Preferred herein are 3-10 membered monocyclic, bicyclic or tricyclic heterocyclyl groups which may contain 1, 2 or 3 ring member atoms selected from nitrogen, oxygen and/or sulfur. Non-limiting examples of "heterocyclyl" include azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, pyrazolidinyl, imidazolidinyl, dioxolanyl, oxathiolanyl, piperidinyl, 2-oxo-piperidinyl, tetrahydropyranyl, thialkyl, piperazinyl, piperazin-2-one, dioxanyl, morpholinyl and thiomorpholinyl, 1-dioxo-thiomorpholinyl, and the like. The heterocyclyl group may be optionally substituted or unsubstituted.
In the definition of "heterocyclyl" a "spiro" refers to a polycyclic ring radical of two or more cyclic structures, with single rings sharing one atom in common with each other, containing several unsaturated bonds within the ring, but no ring having a completely conjugated pi-electron aromatic system, wherein one or more ring members are selected from nitrogen, oxygen or S (O) n (wherein n is selected from 0, 1 or 2) and the remaining ring members are carbon. Preferably 6-12 membered, more preferably 8-9 membered, are used herein. The spiro ring is classified into a single spiro heterocyclic group, a double spiro heterocyclic group or a multiple spiro heterocyclic group according to the number of common spiro atoms between rings, preferably a single spiro heterocyclic group and a double spiro heterocyclic group, more preferably a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered single spiro heterocyclic group. Non-limiting examples thereof include, but are not limited to, 7-azaspiro [3.5 ]]Nonane, 2, 7-diazaspiro [3.5 ]]Nonane, 2-azaspiro [3.4 ]]Octane.
In the definition of "heterocyclyl" a "bridged ring" refers to a polycyclic ring group containing two or more cyclic structures and sharing two or more atoms with each other, one or more of the rings may contain several unsaturated bonds, but none of the ringsAromatic systems having fully conjugated pi electrons, wherein one or more ring members are selected from nitrogen, oxygen or S (O) n (wherein n is selected from 0, 1 or 2) and the remaining ring members are carbon. Preferably 6-12 membered, more preferably 8-9 membered, are used herein. Heterocyclic groups which may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged according to the number of constituent rings, are preferably bicyclic or tricyclic.
In the definition of "heterocyclyl", a "fused ring" refers to a polycyclic ring radical containing two or more cyclic structures and sharing a pair of atoms with each other, one or more of the rings may contain several unsaturated bonds, while at least one ring has a fully conjugated pi-electron aromatic system in which one or more of the ring members is selected from nitrogen, oxygen or S (O) n (wherein n is selected from 0, 1 or 2) and the remaining ring members are carbon. Preferably 6-12 membered, more preferably 8-9 membered, are used herein. The number of constituent rings may be classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group.
The term "heterocycloalkyl" as used herein, alone or in combination, refers to a saturated monocyclic, bicyclic or polycyclic ring radical in which one or more (such as one, two, three or four) ring members are heteroatoms, which may be spiro or bridged. Preferred herein are 3-12 membered cycloalkyl groups, more preferred are 3-10 membered cycloalkyl groups, and most preferred are 3-8 membered cycloalkyl groups. Non-limiting examples of monocyclic heterocycloalkyl groups include, but are not limited to, propylene oxide, thiirane, aziridine, azetidine, oxetane, thietane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, oxazolidine, thiazolidine, imidazolidine, tetrahydropyran, piperidine, dioxane, azepane.
The term "heterocycloalkenyl" as used herein, alone or in combination, refers to a non-aromatic monocyclic, bicyclic or polycyclic saturated heterocyclyl group having 1 or more unsaturated double bonds in which one or more (such as one, two, three or four) ring members are heteroatoms, which may be spiro or bridged. Preferred herein are 3-12 membered heterocycloalkenyl groups, more preferred are 3-10 membered heterocycloalkenyl groups, and most preferred are 3-8 membered heterocycloalkenyl groups. Non-limiting examples of heterocycloalkenyl groups include, but are not limited to, aziridine, oxetane, thiolene, oxetane, pyran.
The term "heteroaryl", as used herein, alone or in combination, refers to a 5-12 membered (preferably 5-10 membered, more preferably 5-6 membered) monocyclic, bicyclic or tricyclic ring system wherein at least one ring is aromatic and at least one ring contains one or more heteroatoms selected from nitrogen, oxygen, sulfur, and wherein said heteroaryl further has one or more attachment points attached to the remainder of the molecule. "heteroaryl" groups of a bicyclic or tricyclic ring system, if they contain saturated or unsaturated heterocycloalkyl groups or groups. Non-limiting examples of "heteroaryl" include furyl, imidazolyl, isoxazolyl, oxazolyl, pyrrolyl, pyrazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, thienyl, thiazolyl, and the like; also included are the following bicyclic rings, but not limited to these: benzimidazolyl, benzofuranyl, benzothienyl, indolyl, oxoindolyl, indolinyl, imidazopyridinyl, pyrazolopyridinyl, pyrazolopyrimidinyl, quinolinyl, isoquinolinyl, quinazolinyl, indazole, 1, 8-naphthyridine, benzo [ d ] isoxazole, benzo [ d ] thiazole, pyrrolo [3,2-b ] pyridine, furan [3,2-b ] pyridine, pyrrolo [1,2-b ] pyridazine, imidazo [1,2-b ] pyridazine, pyrazolo [1,5-a ] pyrimidine, thiazolo [4,5-c ] pyridine, thieno [3,2-b ] pyridine, pyrrolo [1,2-b ] pyridazine, 2, 3-dihydrobenzofuran, benzo [ c ] [1,2,5] oxadiazole, 1, 3-dihydro-2H-benzo [ d ] imidazol-2-one, benzo [ d ] oxazol-2 (3H) -one, and the like. Heteroaryl groups may be optionally substituted or unsubstituted.
The term "halogen" as used herein, alone or in combination, refers to fluorine, chlorine, bromine or iodine.
The term "hydroxy", as used herein, alone or in combination, refers to-OH.
The term "cyano", as used herein, alone or in combination, refers to-CN.
The term "methylsulfonyl" as used herein, alone or in combination, refers to-S (O) 2 -CH 3 。
The term "(substituted) or" substituted with … … "as used herein means that one or more hydrogens on a particular atom are replaced with a specific group (e.g., halogen, alkyl, etc.), where the normal valence of the specified atom is not exceeded under the present circumstances, then the result is a stable compound.
The term "pharmaceutically acceptable salt" as used herein is well known to those skilled in the art.
The term "pharmaceutically acceptable" as used herein refers to a material (e.g., carrier or diluent) that does not affect the biological activity or properties of the compounds of the present invention, and is relatively non-toxic, i.e., the material can be administered to an individual without causing an adverse biological reaction or interacting in an adverse manner with any of the components contained in the composition.
The term "pharmaceutical composition" as used herein refers to a biologically active compound optionally admixed with at least one pharmaceutically acceptable chemical ingredient including, but not limited to, carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents and/or excipients.
The term "carrier" as used herein refers to a relatively non-toxic chemical compound or agent that facilitates the introduction of the compound into a cell or tissue.
The term "stereoisomers" as used herein includes, but is not limited to, enantiomers, cis-trans isomers, and the like.
The term "enantiomer" as used herein refers to a compound having the same formula, in which two compounds which are enantiomers are mirror images of each other and cannot coincide, due to isomerism caused by differences in the spatial configuration of atoms or groups of atoms (groups). The term "cis-trans isomer" as used herein generally refers to a stereoisomerism of diastereomers in a compound molecule that occurs due to the restriction factor of free rotation, which varies the spatial arrangement of the individual groups. Organic molecules containing such isomers, such as olefins, azo compounds, alicyclic hydrocarbons, etc., are considered cis-trans isomerism. In this application, cis-trans isomerism is predominantly embodied in the form of alicyclic hydrocarbons. For example, in cyclohexane, cis-trans isomerism occurs when cyclohexane is substituted with two substituents, the two substituents being the "cis" isomer on the same side of the ring and the "trans" isomer on the different side.
The compounds of the invention may contain asymmetric or chiral centers and thus exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers, sterically hindered isomers and geometric (conformational) isomers and mixtures thereof, such as racemic mixtures, are within the scope of the present invention.
Unless otherwise indicated, structures described herein also include all isomers of such structures (e.g., diastereomers, enantiomers, cis-trans isomers, sterically hindered isomers, geometrically (conformational) isomeric forms), e.g., R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, cis-trans isomers of aliphatic cyclic hydrocarbons, sterically hindered isomers of biphenyl structures (see basic organic chemistry (second edition) upper book, xing Jiyi, etc., p 104-105); PAC,1996,68,2193, (Basic terminology of stereochemistry (IUPAC Recommendations 1996,on page 2201)), (Z) and (E) conformational isomers. Thus, individual stereoisomers, mixtures of enantiomers, mixtures of diastereomers, mixtures of sterically hindered isomers and mixtures of geometric (conformational) isomers of the compounds of the invention are all within the scope of the invention.
The invention will be further elucidated by means of some specific embodiments. For example:
in formula I, the optional groups for each group may be:
in some specific embodiments, ring a is a 5-10 membered heteroaryl. Wherein heteroaryl groups may be in a monocyclic or fused ring form. Ring A is linked to ring B via covalent bonds, respectively, and via L 1 And R is R 1 Is linked and optionally is substituted with 1-3R 3 And (3) substitution. If not specified, R 3 Can be attached to ring A by H atoms substituted for any ring member atom of ring AIncluding C, N.
When ring A is a single ring, more preferably a 6 membered heteroaryl group, the number of heteroatoms is preferably 1 to 3, and the heteroatoms are preferably N. Non-limiting examples thereof include, but are not limited to, pyridine, pyrazine, pyrimidine, pyridazine, triazine. If not specified, on ring A, rings B and L 1 The linking site of (c) may be ortho, meta or para.
When ring A is a fused ring, it is preferably 5-6 fused ring or 6-6 fused ring and forms a conjugated bicyclic aryl, the number of heteroatoms is preferably 1-3, said heteroatoms preferably being N. If not specified, L 1 And ring B are connected to different ones of the parallel rings a, respectively. When ring B is heterocycloalkyl, it preferably contains 1 heteroatom optionally from N, O, S. Non-limiting examples of ring a at this time include, but are not limited to, the following structures:
Typically, the radical R substituted on ring A 3 Non-limiting examples of (C) include, but are not limited to, halogen, hydroxy, nitro, cyano, amino, C 1-6 Alkyl groups, preferably cyano, amino, methyl.
In some embodiments, L 1 Connecting rings A and R by covalent bonds 1 To which are attached, or L 1 Is itself a chemical bond. L (L) 1 Non-limiting examples of (C) include, but are not limited to, imino, carbonyl, -CO-NH-,
In some embodiments, R 1 Selected from C 1-6 Alkyl, 5-10 membered cycloalkyl, 5-10 membered heterocycloalkyl, and optionally substituted with 1-3R 5 Substitution wherein cycloalkyl or heterocycloalkyl includes, but is not limited to, monocyclic, fused, spiro, bridged ring forms, preferably 4-6 membered cycloalkyl or 4-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, S. When R is 1 In the case of cycloalkyl or heterocycloalkyl, preference is given to a 5-to 7-membered monocyclic form, or to a 7-to 9-membered bridged or spiro ringForm of the invention. Wherein the 7-9 membered spiro form has a structure similar to that ofThe structure, dotted line, represents a 4-6 membered cycloalkyl group (including a spiro atom) attached to a spiro atom, any atom may be replaced with a heteroatom, as appropriate to the rules of the current valence of the compound. A 7-9 membered bridged ring form, i.e., where two non-adjacent atoms are attached to each other at the ring group by 1 covalent bond or 1 or more atoms. R is R 1 Non-limiting examples of (a) include, but are not limited to, cyclobutane, cyclopentyl, cyclohexyl, cycloheptyl,/-> Methyl, ethyl, propyl, isopropyl.
When R is 1 Is an asymmetric group, or R 5 Is not at R 1 Different stereoisomers may occur, including enantiomers and cis-trans isomers. It should be noted that both enantiomer and cis-trans isomer have ALK2 inhibitory activity, but different isomer inhibitory activities may be different.
In some embodiments, R 5 Is OH, halogen or C 1-6 Alkyl, - (CR) a R b ) n -OH. Wherein R is a 、R b Preferably H or C 1-3 Alkyl, n is preferably 1 to 3.R is R 5 Non-limiting examples of (C) include, but are not limited to, hydroxy, F, cl, methyl, ethyl, propyl, isopropyl, -CH 2 OH、-C(CH 3 ) 2 OH。
In some embodiments, ring B is a 6-15 membered ring group, and optionally 1-3 ring carbon atoms are replaced with oxygen; ring B is optionally substituted with 1 to 3R 4 And (3) substitution. Ring B may be in the form of a single ring, a double ring or a triple ring, and when ring B is a double ring or a triple ring, the connection form between the rings may be a parallel ring, a spiro ring or a bridged ring. Ring B, unless otherwise specifiedAt least 1 ring of the two groups is an aromatic ring or a heteroaromatic ring, the other rings are saturated or unsaturated cycloalkyl or heterocycloalkyl, or the other rings are aromatic rings or heteroaromatic rings in parallel. If not specified, R 4 The ring B may be attached by an H atom substituted for any of the ring B ring members, including C, N.
When ring B is a single ring, more preferably a 6-membered aryl or heteroaryl group, the number of heteroatoms is preferably 0 to 3, and the heteroatoms are preferably N. Non-limiting examples thereof include, but are not limited to, benzene, pyridine, pyrazine, pyrimidine, pyridazine, triazine. If not specified, on ring B, rings A and L 2 The linking site of (c) may be ortho, meta or para.
When ring B is a parallel ring, it isIn the form, preferably 5-6 or 6-6, Y 1 、Y 2 、Y 3 Independently selected from CH or N. Wherein ring C is a 5-6 membered cyclic group, preferably cycloalkyl, heterocycloalkyl, aryl or heteroaryl in parallel. Ring C optionally contains 1 to 3N atoms and wherein carbon atoms are optionally oxo to form a carbonyl group. If not specified, L 2 And ring a is connected to a different ring of the parallel rings B, respectively.
When ring B isWhen, typically, the connection L is connected by the terminal connection ring a 2 。
When ring B is also a tricyclic group, one of the rings is a benzene ring. In some embodiments, ring B isr, p and q are integers of 1-3; z is Z 1 、Z 2 、Z 3 Independently selected from CH or N, and up to one is N; />Is a single bond or a double bond; when- >Z is a single bond 4 Is CH 2 Or O, Z 5 Is CH 2 Or NH; when->In the case of double bonds, Z 4 Is CH, Z 5 CH or N; optionally 1 ring carbon atom in ring B is replaced by oxygen; ring B is optionally substituted with 1-2R 4 And (3) substitution. In a further preferred embodiment, Z 5 Is limited to CH 2 Or CH, Z 6 Is defined as N and is equal to L 2 And (3) connecting, wherein p is 2. In some more specific embodiments, ring B isr is an integer from 1 to 2, < >>Is a single bond or a double bond.
When ring B isOr further +.>When, typically, the connection L is connected by the terminal connection ring a 2 。
Non-limiting examples of ring B include, but are not limited to, the following structures:
typically, the 6-membered aromatic or heteroaromatic ring is attached(i.e.the bond above ring B) is attached to ring A, the other +.>(i.e., bond under ring B) and L 2 Are connected.
Typically, the radical R substituted on ring B 4 Non-limiting examples of (C) include, but are not limited to, halogen, hydroxy, nitro, cyano, amino, C 1-6 Alkyl groups, preferably cyano, methyl, trifluoromethyl.
In some embodiments, L 2 Connecting rings B and R by covalent bonds 2 Or L 2 Is itself a chemical bond. In some embodiments, L 2 Is defined as- (CR) a R b ) n -, wherein R is a 、R b Independently selected from H or C 1-3 Alkyl, or R a 、R b Forms a 3-6 membered cycloalkyl group with the attached carbon atom. n can be an integer of 1-3, when n is 2, L2 is-CR a R b -CR a ’R b ' at this time R a 、R b 、R a ’、R b ' -can be independently selected from H or C 1-3 Alkyl, while R a And R is R b Or R a ' and R b ' carbon atoms which may be co-linked therewith form a 3-6 membered cycloalkyl group, e.gIn the present application, L 2 Non-limiting examples of (a) include, but are not limited to, a bond, methylene,/->
In some embodiments, R 2 Optionally by 1-2R 6 Substituted, wherein heterocycloalkyl includes, but is not limited to, monocyclic, spiro forms, preferably 4-6 membered cycloalkyl or 4-6The member contains 1-2 heterocycloalkyl groups selected from N, O, S heteroatoms. . When R is 2 In the case of heterocycloalkyl, the 5-7 membered monocyclic form, or the 7-9 membered bridged or spiro form is preferred. Wherein the 7-9 membered spiro form has a structure similar to that ofThe structure, dotted line, represents a 4-6 membered cycloalkyl group (including a spiro atom) attached to a spiro atom, any atom may be replaced with a heteroatom, as appropriate to the rules of the current valence of the compound. A 7-9 membered bridged ring form, i.e., where two non-adjacent atoms are attached to each other at the ring group by 1 covalent bond or 1 or more atoms. R is R 2 Non-limiting examples of (a) include, but are not limited to, cyclopentyl, cyclohexyl, cycloheptyl,/->
When R is 2 Is an asymmetric group, or R 6 Is not at R 2 Different stereoisomers may occur, including enantiomers and cis-trans isomers. It should be noted that both enantiomer and cis-trans isomer have ALK2 inhibitory activity, but different isomer inhibitory activities may be different.
In some embodiments, R 6 Selected from hydroxy, -CO-R c 、-CO-NR c R d 、-SO 2 -R c Or R is 6 Selected from C 1-6 Alkyl, 4-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, optionally substituted with 1-3R e And (3) substitution. Wherein R is c 、R d Independently selected from H, halogen, hydroxy, nitro, cyano, amino or C 1-6 An alkyl group; r is R e Selected from halogen, hydroxy, nitro, cyano, amino, C 1-6 An alkoxy group. R is R 6 Non-limiting examples of (a) include, but are not limited to H, F, methyl, ethyl, isopropyl, hydroxy,
Some compounds of the present application may be classified into a parallel-ring compound and a tricyclic compound according to ring B. These compounds have better ALK2 inhibitory activity and cellular activity and have better selectivity than existing compounds.
Scheme one: parallel ring compound
/>
In this scheme, ring B has the following structure:wherein Y is 1 、Y 2 、Y 3 Independently selected from CH or N, and ring C is a 5-6 membered ring group in parallel. In this scheme, ring C may be a 5-6 membered saturated or unsaturated cycloalkyl or heterocycloalkyl, as well as aryl or heteroaryl. And when ring C is saturated or unsaturated cycloalkyl, heterocycloalkyl, ring C ring member may be substituted with oxygen to form carbonyl. When ring B is->When, typically, the connection L is connected by the terminal connection ring a 2 。
In some more preferred cases, ring B has the following structure:wherein ring C may further preferably be a 5-membered cycloalkyl, nitrogen-containing heterocycloalkyl, nitrogen-containing heteroaryl group in parallel, e.g.>
At this time, R 1 It can be 4-6 membered cycloalkyl or heterocycloalkyl containing 1-2 hetero atoms selected from N, OFor example, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, tetrahydropyranyl, hexahydropyridinyl, morpholin; further, R 1 Optionally substituted with 1-2R 5 Substituted, R 5 Can be hydroxy, halogen or C 1-3 Alkyl, -CH 2 OH、-C(CH 3 ) 2 OH. The scheme is not limited to L 1 R is R 5 At R 1 The linking sites on the substrate, and thus the two linking sites may be ortho, meta or para, preferably meta or para. Further, according to R 1 Different substituents and substitution sites, may exist in stereoisomers, including cis-trans isomers and enantiomers. In general, R 1 Is thatR 5 Is hydroxy or unsubstituted.
In this scheme, L 2 Selected from a bond or a simple (alkylene) group, to which R is attached 2 Ring B. R is R 2 Typically 5-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, optionally substituted with 1-2R 6 Substitution, e.g. R 2 May be L 2 By substitution of R 2 To hydrogen atoms by way of attachment to R 2 Any one of the sites. R is R 6 May be a simple substituent similar to halogen, hydroxy, amino, cyano, or may be optionally substituted with R e Substituted cycloalkyl of 4-7 members, heterocycloalkyl of 4-7 members containing 1-2 hetero atoms selected from N, O, C 1-3 An alkyl group. The scheme is not limited to L 2 R is R 6 At R 4 The linking sites on the substrate, and thus the two linking sites may be ortho, meta or para, preferably meta or para. Further, according to R 2 Different substituents and substitution sites, may be stereoisomerically, including cisTrans-isomerism and enantiomer. R is R e Typically halogen, but also other simple substituents like hydroxy, amino or cyano, also depending on the substitution site and R 6 Stereoisomers may be formed, including cis-trans isomers and enantiomers. />
Scheme II: tricyclic compounds
In this scheme, ring B has the following structure:r, p and q are integers of 1-3; z is Z 1 、Z 2 、Z 3 Independently selected from CH or N, and up to one is N; />Is a single bond or a double bond; when->Z is a single bond 4 Is CH 2 Or O, Z 5 Is CH 2 Or NH; when->In the case of double bonds, Z 4 Is CH, Z 5 CH or N; z is Z 6 Is NH or O; optionally 1 ring carbon atom in ring B is replaced by oxygen. The ring C atom of ring B may be substituted with oxygen to form a carbonyl group.
In some preferred cases, ring B isr or q is an integer from 1 to 3; z is Z 1 、Z 2 Or Z is 3 Independently selected from CH or N, and up to one is N; />Is a single bond or a double bond; when->Z is a single bond 4 Is CH 2 Or O; when->Z is a double bond 4 CH; ring B is optionally substituted with 1-2R 4 Substitution;
in some more preferred cases, ring B has the following structure:wherein r is 1 or 2, e.g When ring B isOr further +.>When, typically, the connection L is connected by the terminal connection ring a 2 。
At this time, R 1 A 4-6 membered cycloalkyl group or a heterocycloalkyl group containing 1 to 2 hetero atoms selected from N, O and the like, for example, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, tetrahydropyranyl, hexahydropyridyl, morpholine; further, R 1 Optionally substituted with 1-2R 5 Substituted, R 5 Can be hydroxy, halogen or C 1-3 Alkyl, -CH 2 OH、-C(CH 3 ) 2 OH. The scheme is not limited to L 1 R is R 5 At R 1 The linking sites on the substrate, and thus the two linking sites may be ortho, meta or para, preferably para. Further, according to R 1 Different substituents and substitution sites, may be stereoisomerically, including cisTrans-isomerism and enantiomer. In general, R 1 Is thatR 5 Is hydroxyl or is not substituted, and the substitution site is para-position substitution.
In this scheme, L 2 Selected from a bond or a simple (alkylene) group, to which R is attached 2 Ring B. R is R 2 Typically 5-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, optionally substituted with 1-2R 6 Substitution, e.g. R 2 May be R 6 Simple substituents of halogen, hydroxy, amino, cyano are preferred, and may be unsubstituted. The scheme is not limited to L 2 R is R 6 At R 4 The linking sites on the substrate, and thus the two linking sites may be ortho, meta or para, preferably para. Further, according to R 2 Different substituents and substitution sites, may exist in stereoisomers, including cis-trans isomers and enantiomers. In general, R 2 Is H or->
Detailed Description
The present invention will be described in further detail with reference to the following examples, but the present invention is not limited to the following examples.
The partial preparation conditions used in the examples are as follows:
preparation of Pre-HPLC conditions: instrument: GILSON-GX281; wavelength: 220nm&254nm; column model: waters X-bridge (30X 100mm,10 μm) or Luna C18 (30X 75mm,3 μm); mobile phase: a:10mM ammonium bicarbonate or H 2 O (0.1% formic acid) or H 2 O (0.1% trifluoroacetic acid), B: acetonitrile; run time: 15min; flow rate: 25mL/min.
Reverse phase column purification was performed using a C18 reverse phase silica gel column (thermal C18,40-60 μm,40g-120 g) with water/acetonitrile (95/5-30/70) as the mobile phase.
Some intermediates of the present invention are synthesized as follows:
intermediate 1: (1R, 5S) -1- (4-bromophenyl) -3- (tetrahydro-2H-pyran-4-yl) -3-azabicyclo [3.1.0] hexane
Synthesis of intermediate 1 reference CN109641871B is directed to the step of synthesizing intermediate A3 from example 34.
Intermediate 2: (1S, 5R) -1- (4-bromophenyl) -3- (tetrahydro-2H-pyran-4-yl) -3-azabicyclo [3.1.0] hexane
Referring to the synthesis of intermediate 1, the first step is to replace (S) -2- (chloromethyl) oxirane with (R) -2- (chloromethyl) oxirane as starting material, and to obtain intermediate 2 via a similar procedure.
Intermediate 3: 6-bromo-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
(1) 6-bromopyrazol [1,5-a ] pyrimidine-3-carboxylic acid
5-amino-1H-pyrazole-4-carboxylic acid (5 g,39 mmol), 2-bromopropionaldehyde (6 g,39 mmol) was dissolved in a mixed solvent of ethanol and acetic acid (10 mL/30 mL). The reaction solution was stirred for 4 hours at 90℃under nitrogen protection. The reaction was cooled, the solid was precipitated, and the title compound (8.5 g, yellow solid) was obtained by filtration, yield: 89%. MS (ESI) m/z 241.9[ M+H ] ] + .
(5) 6-bromo-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
The 6-bromopyrazole [1,5-a ] is reacted with]Pyrimidine-3-carboxylic acid (50 mg,0.2 mmol) was dissolved in N, N-dimethylformamide (1 mL) and 4-aminobicyclo [2.2.2]Octane-1-ol hydrochloride (37 mg,0.2 mmol), HATU (152 mg,0.4 mmol) and N, N-diisopropylethylamine (77 mg,0.6 mmol) were stirred overnight at room temperature. The reaction was purified by Prep-HPLC to give the title compound (60 mg, white solid), yield: 80%. MS (ESI) m/z 364.9[ M+H ]] + .
Intermediate 4: 2-amino-5-bromo-N- (tetrahydro-2H-pyran-4-yl) nicotinamide
To a solution of 2-amino-5-bromonicotinic acid (1.00 g,4.61 mmol) in N, N-dimethylformamide (10 mL) was added HATU (1.93 g,5.07 mmol) and N, N-diisopropylethylamine (714mg, 5.53 mmol) in this order at 20deg.C. The reaction solution was stirred at 20℃for 0.5 hour, and tetrahydro-2H-pyran-4-amine (699 mg,6.91 mmol) was added. The reaction solution was stirred at 20℃for 12 hours. The reaction was diluted with water (40 mL) and extracted with ethyl acetate (40 mL. Times.3). The organic phase was washed with saturated brine (30 mL) and concentrated under reduced pressure to give the title compound (1.00 g, white solid). MS (ESI) m/z 300.0[ M+H ]] + .
Intermediate 5: 2-amino-5-bromo-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) nicotinamide
2-amino-5-bromonicotinic acid (50 mg, 230.39. Mu. Mol) was dissolved in N, N-dimethylformamide (1.0 mL), and HATU (96 mg, 253.43. Mu. Mol) and N, N-diisopropylethylamine (89 mg, 691.18. Mu. Mol) were added sequentially. The reaction mixture was stirred at 20℃for 30 minutes, and 4-aminobicyclo [2.2.2] was added thereto]Octane-1-diol hydrochloride (40 mg,230.39 umol). Finally, the reaction mixture was stirred at 20℃for 12 hours. To the reaction mixture was added brine (10 mL), and extracted with ethyl acetate (10 mL. Times.3). The organic phase was washed with brine (10 mL) and concentrated under reduced pressure to give the title compound (50 mg, orange solid) which was used directly in the next step. MS (ESI) m/z 340.1[ M+H ]] + .
Example 1
Compound 1: 3-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -6- (4- ((1 r,5 s) -3- (tetrahydro-2H-pyran-4-yl) -3-azabicyclo [3.1.0] hexane-1-yl) phenyl) pyrazine-2-carboxamide
(1) 3-amino-6-bromo-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) pyrazine-2-carboxamide
3-amino-6-bromopyrazine-2-carboxylic acid (50 mg, 229.35. Mu. Mol) was dissolved in N, N-dimethylformamide (1 mL), and N, N-diisopropylethylamine (88.92 mg, 688.05. Mu. Mol) and HATU (95.93 mg, 252.28. Mu. Mol) were added sequentially. After the reaction solution was stirred at 20℃for 30 minutes, 4-aminobicyclo [2.2.2] was added to the reaction system]Octane-1-ol hydrochloride (40.75 mg,229.35 mol) and the reaction was stirred at 20℃for 16 hours. The reaction solution was slowly poured into saturated ammonium chloride solution (3 mL) and extracted with ethyl acetate (2 mL x 3). The combined organic phases were washed with saturated brine (3 ml x 2), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product. Purification of the crude product by Prep-TLC (petroleum ether/ethyl acetate=0/1) gave the title compound (30 mg, white solid), yield: 38.34%. MS (ESI) m/z 341.1[ M+H ] ] + .
(2) (1R, 5S) -3- (tetrahydro-2H-pyran-4-yl) -1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) -3-azabicyclo [3.1.0] hexane
Intermediate 1 (0.05 g, 155.16. Mu. Mol) was dissolved in 1, 4-dioxane (0.5 mL) and pinacol biborate (47.28 mg, 186.20. Mu. Mol), potassium acetate (45.68 mg, 465.49. Mu. Mol) and Pd (dppf) Cl were added sequentially 2 (12.67 mg,15.52 umol). The system was replaced with nitrogen 3 times and then warmed to 80 ℃ and stirred for 12 hours. The reaction was cooled to room temperature, diluted with ethyl acetate (5 mL), filtered through celite, the filter cake was washed with ethyl acetate (3 mL x 2), and the filtrate was concentrated under reduced pressure to give the crude product. Purification of the crude product by Prep-TLC (petroleum ether/ethyl acetate=0/1) gave the title compound (0.03 g, pale yellow solid), yield: 52.35%. MS (ESI) m/z 370.3[ M+H ]] + .
(3) 3-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -6- (4- ((1 r,5 s) -3- (tetrahydro-2H-pyran-4-yl) -3-azabicyclo [3.1.0] hexane-1-yl) phenyl) pyrazine-2-carboxamide
(1R, 5S) -3- (tetrahydro-2H-pyran-4-yl) -1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) -3-azabicyclo [3.1.0]Hexane (0.03 g, 81.23. Mu. Mol) was dissolved in a mixed solution of 1, 4-dioxane (3 mL) and water (0.3 mL), and 3-amino-6-bromo-N- (4-hydroxy-bicyclo [ 2.2.2.2) was added sequentially ]Octyl-1-yl) pyrazine-2-carboxamide (0.03 g,87.92 umol), potassium carbonate (33.68 mg,243.70 umol) and Pd (dppf) Cl 2 (5.94 mg,8.12 umol). The system was replaced with nitrogen 3 times and then warmed to 100 ℃ and stirred for 12 hours. The reaction was cooled to room temperature, diluted with ethyl acetate (5 mL), filtered through celite, the filter cake was washed with ethyl acetate (3 mL x 2), and the filtrate was concentrated under reduced pressure to give the crude product. Purification of the crude product by Prep-HPLC gave the title compound (9.0 mg, brown solid), yield: 20.79%. MS (ESI) m/z 504.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.79(s,1H),7.94-7.84(m,3H),7.64-7.53(m,2H),7.24(d,J＝7.7Hz,2H),4.36(s,1H),3.93-3.73(m,2H),3.44-3.36(m,1H),3.31-3.23(m,2H),3.09(d,J＝7.7Hz,1H),2.68-2.55(m,1H),2.46(d,J＝5.6Hz,1H),2.38-2.30(m,1H),2.10-2.03(m,6H),1.90-1.71(m,3H),1.69-1.62(m,6H),1.44-1.28(m,3H),0.88-0.65(m,1H).
Examples 2 to 8
Preparation of Compounds 2 to 8 by analogy with the procedure of example 1, corresponding starting materials and intermediates were used for the synthesis.
Example 9
Compound 9: n- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -6- (4- ((1R, 5S) -3- (tetrahydro-2H-pyran-4-yl) -3-azabicyclo [3.1.0] hexane-1-yl) phenyl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
(1R, 5S) -3- (tetrahydro-2H-pyran-4-yl) -1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) -3-azabicyclo [3.1.0]Hexane (0.03 g, 81.23. Mu. Mol, example 1, step 2) was dissolved in a mixed solution of 1, 4-dioxane (3 mL) and water (0.3 mL), and intermediate 3 (0.035 g, 95.83. Mu. Mol), potassium carbonate (33.68 mg, 243.70. Mu. Mol) and Pd (dppf) Cl were added in this order 2 (5.94 mg,8.12 umol). The system was replaced with nitrogen 3 times and then warmed to 100 ℃ and stirred for 12 hours. The reaction was cooled to room temperature, diluted with ethyl acetate (5 mL), filtered through celite, the filter cake was washed with ethyl acetate (3 mL x 2), and the filtrate was concentrated under reduced pressure to give the crude product. Purification of the crude product by Prep-HPLC gave the title compound (7 mg, pale yellow solid), yield: 16.10%. MS (ESI) m/z 528.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.62(d,J＝2.0Hz,1H),9.17(d,J＝2.0Hz,1H),8.52(s,1H),7.80(d,J＝8.2Hz,2H),7.59(s,1H),7.31(d,J＝8.2Hz,2H),4.36(s,1H),3.93-3.77(m,2H),3.41(d,J＝8.5Hz,1H),3.28(brs,2H),3.10(d,J＝8.6Hz,1H),2.59(d,J＝8.2Hz,1H),2.47-2.43(m,1H),2.36-2.30(m,1H),2.10-2.02(m,6H),1.90(td,J＝7.6,3.9,Hz,1H),1.85-1.73(m,2H),1.70-1.62(m,6H),1.39-1.34(m,3H),0.84-0.76(m,1H).
Example 10
Compound 10: 2-amino-N- ((3 r,6 s) -6- (2-hydroxypropan-2-yl) tetrahydro-2H-pyran-3-yl) -5- (4- ((1 r,5 s) -3- (tetrahydro-2H-pyran-4-yl) -3-azabicyclo [3.1.0] hexane-1-yl) phenyl) nicotinamide
(1) 2-amino-5-bromo-N- ((3 r,6 s) -6- (2-hydroxy-propan-2-yl) tetrahydro-2H-pyran-3-yl) nicotinamide
2-amino-5-bromonicotinic acid (143 mg, 659. Mu. Mol), HATU (183mg, 483. Mu. Mol) and N, N-diisopropylethylamine (198mg, 1.54 mmol) were dissolved in N, N-dimethylformamide (0.7 mL). The reaction solution was stirred at 20℃for 30 minutes, and 2- ((2S, 5R) -5-aminotetrahydro-2H-pyran-2-yl) propan-2-ol (70.0 mg,439 umol) was added thereto. The reaction solution is continued inStirring is carried out at 20℃for 12 hours. The reaction was filtered and the filtrate purified by Prep-HPLC to give the title compound (60.0 mg, white solid), yield: 38.1%. MS (ESI) m/z 358.1[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.25(d,J＝7.7Hz,1H),8.15(d,J＝2.4Hz,1H),8.09(d,J＝2.3Hz,1H),7.22(s,2H),4.24(s,1H),3.94-3.77(m,2H),3.10(t,J＝10.5Hz,1H),2.97(dd,J＝11.1,1.5Hz,1H),1.96(d,J＝11.7Hz,1H),1.80(d,J＝12.3Hz,1H),1.56-1.51(m,1H),1.43-1.30(m,1H),1.09(s,3H),1.04(s,3H).
(2) 2-amino-N- ((3 r,6 s) -6- (2-hydroxypropan-2-yl) tetrahydro-2H-pyran-3-yl) -5- (4- ((1 r,5 s) -3- (tetrahydro-2H-pyran-4-yl) -3-azabicyclo [3.1.0] hexane-1-yl) phenyl) nicotinamide
2-amino-5-bromo-N- ((3R, 6S) -6- (2-hydroxy-propan-2-yl) tetrahydro-2H-pyran-3-yl) nicotinamide (69.8 mg,194 umol) and (1R, 5S) -3- (tetrahydro-2H-pyran-4-yl) -1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) -3-azabicyclo [3.1.0]Hexane (60.0 mg, 162. Mu. Mol, example 1, step 2) was dissolved in a mixed solvent of 1, 4-dioxane (0.6 mL) and water (0.12 mL), and potassium carbonate (44.9 mg, 324. Mu. Mol) was added. The reaction system was replaced three times with nitrogen, and Pd (dppf) Cl was added to the reaction solution under the protection of nitrogen 2 (11.8 mg,16.2 umol). The reaction solution was stirred for 12 hours at 80 ℃. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated. Purification of the crude product by Prep-HPLC gave the title compound (33.6 mg, brown solid), yield: 38.07%. MS (ESI) m/z 521.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.39(d,J＝2.3Hz,1H),8.29(d,J＝7.8Hz,1H),8.13(d,J＝2.1Hz,1H),7.58(d,J＝8.3Hz,2H),7.23(d,J＝8.3Hz,2H),7.13(s,2H),4.26(s,1H),3.98-3.81(m,4H),3.39(d,J＝8.5Hz,1H),3.31-3.25(m,2H),3.18-3.06(m,2H),2.99(d,J＝10.1Hz,1H),2.56(d,J＝8.6Hz,2H),2.35-2.27(m,1H),2.04-1.97(m,1H),1.88-1.71(m,4H),1.59-1.57(m,1H),1.46-1.33(m,3H),1.31(t,J＝3.9Hz,1H),1.11(s,3H),1.05(s,3H),0.76(dd,J＝7.6,3.4Hz,1H).
Examples 11 to 14
Preparation of Compounds 11 to 14 by analogy with the procedure of example 10, corresponding starting materials and intermediates were used for the synthesis.
Example 15
Compound 15:6- (4- (4- (dimethylcarbamoyl) piperazin-1-yl) phenyl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
(1) 4- (4-bromophenyl) -N, N-dimethylpiperazine-1-carboxamide
1- (4-bromophenyl) piperazine (1 g,4.1 mmol) and triethylamine (8238 mg,8.2 mmol) were dissolved in dichloromethane (5 mL). Dimethylcarbamoyl chloride (538 mg,4.9 mmol) was added dropwise thereto at 0℃and the reaction was stirred at room temperature overnight. Water (20 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (40 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (100% ethyl acetate) to give the title compound (1.2 g, white solid), yield: 92%. MS (ESI) m/z 311.9[ M+H ]] + .
(2) N, N-dimethyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) piperazine-1-carboxamide
4- (4-bromophenyl) -N, N-dimethylpiperazine-1-carboxamide (100 mg,0.32 mmol) and pinacol biborate (90 mg,0.35 mmol) were dissolved in 1, 4-dioxane (5 mL), pdCl was added 2 (dppf) (25 mg,0.03 mmol) and potassium acetate (63 mg,0.64 mmol), the reaction was stirred at 100deg.C under nitrogen overnight. The reaction was filtered, concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (40 mg, yellow solid), yield: 35%. MS (ESI) m/z 360.2[ M+H ]] + .
(3) 6- (4- (4- (dimethylcarbamoyl) piperazin-1-yl) phenyl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
N, N-dimethyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl)) Phenyl) piperazine-1-carboxamide (40 mg,0.11 mmol) and intermediate 3 (40 mg,0.11 mmol) were dissolved in a mixed solvent of 1, 4-dioxane and water (10 mL/2 mL), and PdCl was added 2 (dppf) (8 mg,0.01 mmol) and potassium carbonate (30 mg,0.22 mmol). The reaction was heated to 100 ℃ under nitrogen and stirred overnight. The reaction solution was dried by spin-drying, dissolved in methanol, filtered, and concentrated under reduced pressure. Purification of the residue by Prep-HPLC gave the title compound (5.9 mg, white solid), yield: 10%. MS (ESI) m/z 518.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.53(d,J＝2.0Hz,1H),9.15(d,J＝2.0Hz,1H),8.48(s,1H),7.77(d,J＝8.8Hz,2H),7.58(s,1H),7.09(d,J＝9.2Hz,2H),4.34(s,1H),3.27-3.25(m,8H),2.79(s,6H),2.07-2.04(m,6H),1.67-1.63(m,6H).
Example 16
Compound 16:4- (4- (5- ((4-hydroxy-bicyclo [2.2.2] oct-1-yl) carbamoyl) pyridin-3-yl) phenyl) -N, N-dimethylpiperazine-1-carboxamide
(1) 5- (4- (4- (dimethylcarbamoyl) piperazin-1-yl) phenyl) nicotinic acid
N, N-dimethyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) piperazine-1-carboxamide (72 mg,0.2 mmol) was dissolved in 1, 4-dioxane (5 mL) solution and Pd (dppf) Cl was added 2 (14.6 mg,0.02 mmol), potassium carbonate (82.8 mg,0.6 mmol), 5-bromonicotinic acid (32.3 mg,0.16 mmol) and water (1 mL). The mixture was reacted for 18 hours under nitrogen protection at 100 ℃. The solvent was removed under reduced pressure, the crude product was dissolved in methanol and filtered, and the filtrate was purified by reverse phase column to give the title compound (24 mg, colorless oil), yield: 42%. MS (ESI) m/z 355.0[ M+H ] ] + .
(2) 4- (4- (5- ((4-hydroxy-bicyclo [2.2.2] oct-1-yl) carbamoyl) pyridin-3-yl) phenyl) -N, N-dimethylpiperazine-1-carboxamide
5- (4- (4- (dimethylcarbamoyl) piperazin-1-yl) phenyl) nicotinic acid (24 mg,0.068 mmol) was dissolved in N, N-dimethylformamide (2 mL), 4-aminobicyclo [2.2.2]Octane-1-hydrochloride (12 mg, 0).068 mmol), HATU (51.7 mg,0.136 mmol) and N, N-diisopropylethylamine (43.9 mg,0.34 mmol). The reaction solution was stirred at room temperature for 2 hours. The reaction was filtered through a filter membrane and the filtrate was purified by Prep-HPLC to give the title compound (13.8 mg, white solid), yield: 42%. MS (ESI) mz 478.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.92(d,J＝2.0Hz,1H),8.78(d,J＝2.0Hz,1H),8.25(t,J＝2.2Hz,1H),7.91(s,1H),7.68(d,J＝8.8Hz,2H),7.08(d,J＝9.2Hz,2H),4.30(s,1H),3.27-3.24(m,8H),2.79(s,6H),2.07-2.03(m,6H),1.65-1.60(m,6H).
Example 17
Compound 17:6- (4- (4- (dimethylcarbamoyl) piperazin-1-yl) phenyl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) pyrazine-2-carboxamide
(1) 6- (4- (4- (dimethylcarbamoyl) piperazin-1-yl) phenyl) pyrazine-2-carboxylic acid
The procedure of step (1) was followed, using 6-bromopyrazine-2-carboxylic acid methyl ester (110 mg,0.5 mmol) as a starting material, to give the title compound (30 mg, green solid), yield: 17.5%. MS (ESI) m/z 356.0[ M+H ]] + .
(2) 6- (4- (4- (dimethylcarbamoyl) piperazin-1-yl) phenyl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) pyrazine-2-carboxamide
Reference to example 16 procedure (2) was followed as 6- (4- (4- (dimethylcarbamoyl) piperazin-1-yl) phenyl) pyrazine-2-carboxylic acid (30 mg,0.08 mmol) and 4-aminobicyclo [ 2.2.2.2]Octane-1-hydrochloride (11 mg,0.08 mmol) as the starting material gave the title compound (13 mg, white solid), yield: 32.5%. MS (ESI) m/z 479.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.33(s,1H),8.92(s,1H),8.13(d,J＝8.8Hz,2H),7.89(s,1H),7.09(d,J＝8.8Hz,2H),4.35(s,1H),3.31-3.26(m,8H),2.79(s,6H),2.11-2.07(m,6H),1.67-1.63(m,6H).
Example 18
Compound 18:6- (4- (1-ethylpiperidin-4-yl) phenyl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
(1) 4- (4-bromophenyl) -1-ethylpiperidine
To a solution of 4- (4-bromophenyl) piperidine (848 mg,3.53 mmol) in N, N-dimethylformamide (15 mL) was added N, N-diisopropylethylamine (1.36 g,10.6 mmol) and iodoethane (661mg, 4.24 mmol), and the mixture was reacted overnight at room temperature. The reaction was poured into water (100 mL), extracted with ethyl acetate (2×40 mL), and the organic phases were combined and washed with saturated brine (2×50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound (540 mg, pale yellow solid), yield: 57%. MS (ESI): m/z 267.9[ M+H ]] + .
(2) 1-ethyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) piperidine
To a solution of 4- (4-bromophenyl) -1-ethylpiperidine (214 mg,0.8 mmol) in 1, 4-dioxane (10 mL) was added Pd (dppf) Cl in sequence 2 (58 mg,0.08 mmol), potassium acetate (235 mg,2.4 mmol) and pinacol biborate (244 mg,0.96 mmol) were heated to 100deg.C under nitrogen and stirred for 18 hours. The solvent was removed under reduced pressure and the crude product was purified by column chromatography (dichloromethane/methanol=20/1) to give the title compound (200 mg, brown oil), yield: 79%. MS (ESI) m/z 316.1[ M+H ]] + .
(3) 6- (4- (1-ethylpiperidin-4-yl) phenyl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
The procedure of step (3) was followed as described in example 15, starting from 1-ethyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) piperidine (41 mg,0.13 mmol) and intermediate 3 (36.5 mg,0.1 mmol) to give the title compound (16.1 mg, white solid) in yield: 34%. MS (ESI) m/z 474.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.60(d,J＝2.4Hz,1H),9.15(d,J＝2.4Hz,1H),8.52(s,1H),7.81-7.79(m,2H),7.58(s,1H),7.42-7.40(m,2H),4.34(s,1H),3.01-3.98(m,2H),2.56-2.53(m,1H),2.36(q,J＝7.2Hz,2H),2.09-2.05(m,6H),2.01-1.95(m,2H),1.80-1.76(m,2H),1.74-1.70(m,1H),1.68-1.64(m,7H),1.03(t,J＝7.2Hz,3H).
Example 19
Compound 19: n- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -6- (4- (1- (pyrrolidin-1-yl) ethyl) phenyl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
(1) 6- (4-Acetylphenyl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
(4-Acetylphenyl) boronic acid (100 mg,0.6 mmol) and intermediate 3 (220 mg,0.6 mmol) were dissolved in a mixed solvent of 1, 4-dioxane and water (10 mL/2 mL), and PdCl was added 2 (dppf) (43 mg,0.06 mmol) and potassium carbonate (165 mg,1.2 mmol), the reaction was stirred overnight under nitrogen at 90 ℃. The reaction was directly filtered and dried, and the residue was purified by reverse phase column to give the title compound (30 mg, yellow solid), yield: 11.6%. MS (ESI) m/z 405.0[ M+H ]] + .
(2) N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -6- (4- (1- (pyrrolidin-1-yl) ethyl) phenyl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
6- (4-Acetylphenyl) -N- (4-hydroxy-bicyclo [ 2.2.2)]Oct-1-yl) pyrazolo [1,5-a]Pyrimidine-3-carboxamide (30 mg,0.07 mmol) and pyrrolidine (10 mg,0.14 mmol) were dissolved in methanol (3 mL) and sodium cyanoborohydride (10 mg,0.17 mmol) was added after stirring the reaction at 50℃for 0.5 h, and the reaction was continued to be stirred overnight at 50 ℃. The reaction was purified by Prep-HPLC to give the title compound (5.4 mg, white solid), yield: 15.9%. MS (ESI) m/z 460.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.61(d,J＝2.4Hz,1H),9.17(d,J＝2.4Hz,1H),8.52(s,1H),7.82(d,J＝10.4Hz,2H),7.58(s,1H),7.47(d,J＝7.6Hz,2H),4.34(s,1H),2.33-2.32(m,3H),2.09-2.05(m,7H),1.68-1.64(m,11H),1.32(d,J＝6.4Hz,3H).
Example 20
Compound 20:6- (5- (4- (dimethylcarbamoyl) piperazin-1-yl) pyridin-2-yl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
/>
(1) 4- (6-bromopyridin-3-yl) piperazine-1-carboxylic acid tert-butyl ester
2-bromo-5-iodopyridine (1.8 g,6.5 mmol), piperazine-1-carboxylic acid tert-butyl ester (1 g,5.4 mmol), sodium tert-butoxide (1.5 g,16 mmol), tris (dibenzylideneacetone) dipalladium (100 mg) and 4, 5-bis (diphenylphosphine) -9, 9-dimethylxanthene (100 mg) were dissolved in toluene (20 mL). The reaction solution was stirred overnight at 90℃under nitrogen. Water (50 mL) was added to the reaction system for dilution, extraction with ethyl acetate (20 mL x 2), drying over anhydrous sodium sulfate, filtration, and concentration under reduced pressure gave the title compound (1.8 g, crude product), yield: 100%. MS (ESI) m/z 341.9[ M+H ] ] + .
(2) 1- (6-bromopyridin-3-yl) piperazine
4- (6-bromopyridin-3-yl) piperazine-1-carboxylic acid tert-butyl ester (1.8 g, crude) was dissolved in methylene chloride (5 mL), and trifluoroacetic acid (5 mL) was added and stirred at room temperature for 2 hours. Concentrating under reduced pressure to give 1- (6-bromopyridin-3-yl) piperazine (1.3 g, crude), yield: 100%. MS (ESI) m/z 241.9[ M+H ]] + .
(3) 4- (6-bromopyridin-3-yl) -N, N-dimethylpiperazine-1-carboxamide
1- (6-bromopyridin-3-yl) piperazine (1.3 g,5.4 mmol) and triethylamine (5.4 g,54 mmol) were dissolved in methylene chloride (10 mL), and dimethylcarbamoyl chloride (870 mg,8.1 mmol) was added thereto, and the reaction mixture was stirred at room temperature for 2 hours. Water (50 mL) was added thereto, the mixture was extracted with ethyl acetate (20 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=1/1) to give the title compound (1.5 g, yellow solid). MS (ESI) m/z 312.9[ M+H ]] + .
(4) N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -6- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
Intermediate 3 (40 mg,0.1 mmol) was dissolved in 1, 4-dioxane (1 mL) and pinacol biborate (33 mg,0.13 mmol), potassium acetate (43 mg0.44 mmol) and Pd (dppf) Cl 2 (50 mg). The reaction was stirred at reflux under nitrogen overnight. Concentrating the reaction solution to obtain N- (4-hydroxy bicyclo [ 2.2.2) ]Oct-1-yl) -6- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazolo [1,5-a]Pyrimidine-3-carboxamide (45 mg, crude), yield: 100%.
(5) 6- (5- (4- (dimethylcarbamoyl) piperazin-1-yl) pyridin-2-yl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
N- (4-hydroxy bicyclo [ 2.2.2)]Oct-1-yl) -6- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazolo [1,5-a]Pyrimidine-3-carboxamide (45 mg,0.1 mmol) was dissolved in a mixed solvent of 1, 4-dioxane and water (5 mL/1 mL), 4- (6-bromopyridin-3-yl) -N, N-dimethylpiperazine-1-carboxamide (41 mg,0.13 mmol), potassium carbonate (45 mg,0.33 mmol) and Pd (dppf) Cl were added 2 (50 mg). The reaction was stirred at reflux under nitrogen overnight. Water (10 mL) was added to the reaction system, extracted with ethyl acetate (5 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification of the residue by Prep-HPLC gave the title compound (1.4 mg, yellow solid), yield: 2.5%. MS (ESI): m/z 519.2[ M+H ]] + . 1 H NMR(400MHz,MeOD-d 4 )δ9.40(d,J＝2.0Hz,1H),9.25(d,J＝2.0Hz,1H),8.41(s,1H),8.40(d,J＝2.8Hz,1H),7.80(d,J＝8.8Hz,1H),7.40(dd,J＝8.8,2.8Hz,1H),3.38-3.26(m,8H),2.81(s,6H),2.14-2.08(m,6H),1.74-1.70(m,6H).
Example 21
Compound 21: n- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -6- (4- (4- (methylsulfonyl) piperazin-1-yl) phenyl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
(1) 1- (4-bromophenyl) -4- (methylsulfonyl) piperazine
1- (4-bromophenyl) piperazine (1 g,4.1 mmol), triethylamine (8238 mg,8.2 mmol) were dissolved in dichloromethane (5 mL), methanesulfonyl chloride (563.5 mg,4.9 mmol) was added dropwise at 0℃and the reaction was warmed to room temperature and stirred overnight. Adding water into the reaction system20 mL), extracted with ethyl acetate (40 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (100% ethyl acetate) to give 1- (4-bromophenyl) -4- (methylsulfonyl) piperazine (1.2 g, white solid), yield: 92%. MS (ESI) m/z 318.9[ M+H ]] + .
(2) 1- (methylsulfonyl) -4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) piperazine
1- (4-bromophenyl) -4- (methylsulfonyl) piperazine (120 mg,0.38 mmol) and pinacol biborate (115 mg,0.45 mmol) were dissolved in 1, 4-dioxane (5 mL) and PdCl was added 2 (dppf) (31 mg,0.04 mmol) and potassium acetate (75 mg,0.76 mmol), the reaction was stirred overnight under nitrogen at 100 ℃. The reaction was filtered, concentrated under reduced pressure, and the residue was purified by Pre-HPLC to give the title compound (15 mg, white solid), yield: 11%. MS (ESI) m/z 367.0[ M+H ]] + .
(3) N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -6- (4- (4- (methylsulfonyl) piperazin-1-yl) phenyl) pyrazolo [1,5-a ] pyrimidine-3-carboxamide
1- (methylsulfonyl) -4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) piperazine (15 mg,0.04 mmol), intermediate 3 (15 mg,0.04 mmol), pdCl 2 (dppf) (4 mg,0.04 mmol), potassium carbonate (11 mg,0.08 mmol) was dissolved in a mixed solvent of 1, 4-dioxane and water (5 mL/1 mL). The reaction solution was stirred overnight at 90℃under nitrogen. The reaction was concentrated under reduced pressure, and the residue was purified by Prep-HPLC to give the title compound (2.7 mg, white solid), yield: 13%. MS (ESI) m/z 525.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.54(d,J＝2.4Hz,1H),9.15(d,J＝2.4Hz,1H),8.49(s,1H),7.78(d,J＝8.8Hz,2H),7.58(s,1H),7.13(d,J＝9.2Hz,2H),4.35(s,1H),3.27-3.25(m,8H),2.93(s,3H),2.08-2.04(m,6H),1.68-1.64(m,6H).
Example 22
Compound 22: (R) -2-amino-5- (4- ((2-methylpyrrolidin-1-yl) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
(1) (R) -1- (4-bromobenzyl) -2-methylpyrrolidine
4-bromobenzaldehyde (500 mg,2.7 mmol) and (R) -2-methylpyrrolidine (230 mg,2.7 mmol) were dissolved in methylene chloride (10 mL), and after stirring at room temperature for half an hour, sodium cyanoborohydride (250 mg,4 mmol) was added thereto, and the reaction system was stirred at room temperature overnight. After completion of the reaction, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate/petroleum ether=1/5) to give the title compound (120 mg, yellow solid), yield: 17.5%. MS (ESI) m/z 254.0[ M+H ]] + .
(2) (R) -2-amino-5- (4- ((2-methylpyrrolidin-1-yl) methyl) phenyl) nicotinic acid
(R) -1- (4-bromobenzyl) -2-methylpyrrolidine (120 mg,0.50 mmol) was dissolved in a mixed solvent of 1, 4-dioxane (10 mL) and water (2 mL), and (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boric acid (98 mg,0.50 mmol) and potassium carbonate (200 mg,0.75 mmol) were added thereto under nitrogen, and [1,1' -bis (diphenylphosphine) ferrocene was added thereto]Palladium dichloride (10 mg,0.03 mmol) was then heated to 90℃under nitrogen and stirred for 12 hours. Water (20 mL) was added to the reaction system, the mixture was extracted with ethyl acetate (20 mL. Times.2), and the organic phase was washed successively with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase column to give the title compound (50 mg, yellow solid), yield: 33%. MS (ESI) m/z 312.1[ M+H ]] + .
(3) (R) -2-amino-5- (4- ((2-methylpyrrolidin-1-yl) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
(R) -2-amino-5- (4- ((2-methylpyrrolidin-1-yl) methyl) phenyl) nicotinic acid (50 mg,0.16 mmol) was dissolved in dichloromethane (5 mL), and tetrahydro-2H-pyran-4-amine (16 mg,0.16 mmol), HATU (91 mg,0.24 mmol), and N, N-diisopropylethylamine (41 mg,0.32 mmol) were added in this order and the reaction stirred at room temperature for 3 hours. The reaction was concentrated and the residue was purified by Pre-HPLC to give the title compound (10 mg, white solid), yield: 15.8%. MS (ESI): m/z 395.2[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.42-8.39(m,2H),8.16(d,J＝4.0Hz,1H),7.60(d,J＝8.0Hz,2H),7.35(d,J＝8.0Hz,2H),7.12(s,2H),4.01-3.96(m,2H),3.91-3.87(m,2H),3.42-3.38(m,2H),3.17-3.13(m,1H),2.80-2.75(m,1H),2.41-2.36(m,1H),2.10-2.03(m,1H),1.92-1.88(m,1H),1.80-1.76(m,2H),1.62-1.53(m,4H),1.40-1.30(m,1H),1.12(d,J＝8.0Hz,3H).
Example 23
Compound 23: 2-amino-5- (4- (morpholinomethyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
(1) 2-amino-5- (4- (morpholinomethyl) phenyl) nicotinic acid methyl ester
4- (4-bromobenzyl) morpholine (100 mg,0.39 mmol), (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boronic acid (76 mg,0.39 mmol), pd (dppf) Cl 2 (29 mg,0.039 mmol) and potassium carbonate (108 mg,0.78 mmol) were dissolved in a mixed solvent of 1, 4-dioxane and water (10 mL, v/v=4/1), nitrogen was substituted, and the reaction system was stirred at 100℃for 16 hours. The reaction was directly stirred and purified by column chromatography (petroleum ether/ethyl acetate=1/1) to give the title compound (70 mg, yellow solid), yield: 55.1%. MS (ESI) m/z 328.0[ M+H ]] + .
(2) 2-amino-5- (4- (morpholinomethyl) phenyl) nicotinic acid
Methyl 2-amino-5- (4- (morpholinomethyl) phenyl) nicotinate (70 mg,0.21 mmol) and lithium hydroxide (24 mg,1.0 mmol) were dissolved in a mixed solvent of methanol and water (5 ml, v/v=3/1), and the reaction system was stirred at room temperature for 3 hours. The reaction solution was directly dried by spin to give the title compound (50 mg, white solid). Yield: 74.6%.
(3) 2-amino-5- (4- (morpholinomethyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
2-amino-5- (4- (morpholinomethyl) phenyl) nicotinic acid (50 mg,0.16 mmol) and tetrahydro-2H-pyran-4-amine (16 mg,0.16 mmol) were dissolved in N, N-dimethylformamide (2 mL) and HATU (76 mg,0.32 mmol) and triethylamine (103 mg,0.80 mmol) were added. The reaction system was stirred at 25℃for 16 hours. The reaction solution was filtered, and the filtrate was concentrated and purified by Pre-HPLC to give the title compound (8.1) mg, white solid), yield: 12.7%. MS (ESI) m/z 397.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.40-8.39(m,2H),8.16(d,J＝2.4Hz,1H),7.61(d,J＝8.0Hz,2H),7.37(d,J＝8.0Hz,2H),7.13(s,2H),4.01-3.98(m,1H),3.91-3.87(m,2H),3.59-3.56(m,4H),3.48(s,2H),3.41-3.36(m,2H),2.37-2.33(m,4H),1.79-1.76(m,2H),1.61-1.55(m,2H).
Example 24
Compound 24:5- (4- ((2-oxa-6-azaspiro [3.3] heptan-6-yl) methyl) phenyl) -2-amino-N- (tetrahydro-2H-pyran-4-yl) nicotinamide
(1) (6-amino-5- ((tetrahydro-2H-pyran-4-yl) carbamoyl) pyridin-3-yl) boronic acid
Intermediate 4 (100 mg,0.33 mmol), pinacol biborate (83 mg,0.33 mmol), pd (dppf) Cl 2 (27 mg,0.033 mmol) and potassium acetate (65 mg,0.66 mmol) were dissolved in 1, 4-dioxane (10 mL), nitrogen was replaced, and the reaction system was stirred at 100deg.C for 16 hours. The reaction solution was used directly in the next step.
(2) 5- (4- ((2-oxa-6-azaspiro [3.3] heptan-6-yl) methyl) phenyl) -2-amino-N- (tetrahydro-2H-pyran-4-yl) nicotinamide
The reaction solution of the previous step is cooled to room temperature, and 6- (4-bromobenzyl) -2-oxa-6-azaspiro [3.3] is added]Heptane (88 mg,0.33 mmol), pd (dppf) Cl 2 (27 mg,0.033 mmol), potassium carbonate (91 mg,0.66 mmol) and water (3 mL), nitrogen was replaced, and the reaction system was stirred at 100deg.C for 16 hours. The reaction was cooled to room temperature, filtered, and the filtrate purified by Pre-HPLC to give the title compound (0.8 mg, white solid), yield: 0.5%. MS (ESI) m/z 409.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.48-8.42(m,2H),8.18(d,J＝2.0Hz,1H),7.83(d,J＝10.8Hz,2H),7.33(d,J＝11.2Hz,2H),7.18(s,2H),4.64(s,4H),4.06-3.91(m,4H),3.53-3.51(m,3H),3.31(s,4H),1.83-1.79(m,2H),1.65-1.58(m,2H).
Example 25
Compound 25: 2-amino-5- (4- (1- (pyrrolidin-1-yl) ethyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
(1) 1- (1- (4-bromophenyl) ethyl) pyrrolidine
To a solution of 1- (4-bromophenyl) ethanone (10.00 g,50.24 mmol) in methanol (100 mL) at 20deg.C was added sequentially tetrahydropyrrole (17.87 g,251.20 mmol) and sodium cyanoborohydride (6.95 g,110.53 mmol). The reaction solution was stirred at 20℃for 12 hours. The reaction solution was concentrated under reduced pressure to remove the solvent, diluted with water (200 mL), and extracted with ethyl acetate (100 mL. Times.2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure to give the title compound (12.5 g, colorless oil). MS (ESI) m/z 254.1[ M+H ]] + .
(2) 1- (1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) ethyl) pyrrolidine
1- (1- (4-bromophenyl) ethyl) pyrrolidine (12.50 g,49.18 mmol) was dissolved in 1, 4-dioxane (125 mL) at 20deg.C under nitrogen, and pinacol biboronate (24.98 g,98.36 mmol), potassium acetate (9.65 g,98.36 mmol) and Pd (dppf) Cl were added sequentially 2 (3.60 g,4.92 mmol). The reaction system was purged with nitrogen three times, followed by stirring at 105℃for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=100/1 to 0/1) to give the title compound (3.00 g, tan solid), yield: 20.2%. MS (ESI) m/z 302.3[ M+H ] ] + .
(3) 2-amino-5- (4- (1- (pyrrolidin-1-yl) ethyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
1- (1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) ethyl) pyrrolidine (30 mg,99.59 umol) was dissolved in a mixed solvent of 1, 4-dioxane (0.15 mL) and water (0.15 mL) under nitrogen protection at 20℃to which intermediate 4 (59 mg,199.18 umol) and potassium carbonate (41 mg,298.77 umol) were added. Pd (dppf) Cl was added under nitrogen blanket 2 (14 mg,19.92 umol). The reaction system was purged with nitrogen three times, then inStirred at 80℃for 12 hours. The reaction solution was cooled to room temperature, diluted with water (5 mL), extracted with ethyl acetate (5 mL), and the organic phase was concentrated under reduced pressure to give the crude product. Purification of the crude product by Prep-HPLC gave the title compound (5 mg, white solid), yield: 12.7%. MS (ESI): m/z 395.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.43-8.35(m,2H),8.16(d,J＝2.3Hz,1H),7.59(d,J＝8.1Hz,2H),7.39(d,J＝8.1Hz,2H),7.14(s,2H),4.07-3.96(m,1H),3.94-3.86(m,2H),3.43-3.36(m,2H),3.25(d,J＝6.1Hz,1H),2.50-2.45(m,2H),2.34(brs,2H),1.78(dd,J＝12.7,1.9Hz,2H),1.73-1.64(m,4H),1.63-1.52(m,2H),1.32(d,J＝6.5Hz,3H).
Example 26
Compound 26: 2-amino-5- (4- (2-morpholinoethoxy) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
4- (2- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) ethyl) morpholine (100 mg,300 mol), intermediate 4 (99.0 mg,330 mol), sodium carbonate (16.5 mg,156 mol) was dissolved in 1, 4-dioxane (1.0 mL) and water (0.1 mL). Pd (dppf) Cl was added to the reaction solution under nitrogen protection 2 (24.62 mg,33.65 umol). The reaction mixture was stirred under nitrogen at 90 ℃ for 16 hours. The reaction was cooled to room temperature, diluted with methanol (1 mL) and filtered, and the filtrate was purified by Prep-HPLC to give the title compound (75.6 mg, pale yellow solid), yield: 59.07%. MS (ESI) m/z 427.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.41(d,J＝7.6Hz,1H),8.36(d,J＝2.4Hz,1H),8.12(d,J＝2.3Hz,1H),7.63-7.54(m,2H),7.07(s,2H),7.03(d,J＝8.7Hz,2H),4.12(t,J＝5.8Hz,2H),4.06-3.95(m,1H),3.90(dd,J＝11.7,2.3Hz,2H),3.62-3.56(m,4H),3.43-3.35(m,2H),2.71(t,J＝5.8Hz,2H),2.50-2.48(m,4H),1.78(dd,J＝12.5,2.2Hz,2H),1.60-1.52(m,2H).
Example 27
Compound 27: 2-amino-5- (4- (1-morpholinocyclopropyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
Referring to the procedure of example 26, starting from 4- (1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) cyclopropyl) morpholine (80.0 mg,242 mol) and intermediate 4 (80.2 mg,267.2 mol), the title compound (28.2 mg, pale yellow solid) was obtained in the yield: 27.47%. MS (ESI) m/z 423.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.42-8.40(m,2H),8.17(d,J＝2.1Hz,1H),7.63(d,J＝8.1Hz,2H),7.33(d,J＝8.1Hz,2H),7.15(s,2H),4.08-3.95(m,1H),3.90(d,J＝9.4Hz,2H),3.50(t,J＝4.1Hz,4H),3.43-3.35(m,2H),2.45-2.43(m,4H),1.83-1.74(m,2H),1.64-1.51(m,2H),0.95-0.88(m,2H),0.80-0.73(m,2H).
Example 28
Compound 28: (R) -2-amino-5- (4- ((3-methylmorpholino) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
(1) (R) -4- (4-bromobenzyl) -3-methylmorpholine
(R) -3-methylmorpholine (1.60 g,15.82 mmol) was dissolved in acetonitrile (16 mL) and potassium iodide (5.25 g,31.64 mmol), potassium carbonate (6.56 g,47.46 mmol) and p-bromobenzyl bromide (4.35 g,17.40 mmol) were added sequentially. The reaction solution was warmed to 60℃and stirred for 12 hours. The reaction solution is cooled to room temperature and filtered, and the filtrate is decompressed and concentrated to obtain crude products. The crude product was purified by column chromatography (petroleum ether/ethyl acetate=100/1 to 0/1) to give the title compound (3.50 g, yellow oil), yield: 81.9%. MS (ESI) m/z 270.2[ M+H ] ] + .
(2) (R) -3-methyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) morpholine
(R) -4- (4-bromobenzyl) -3-methylmorpholine (800 mg,2.96 mmol) was dissolved in N, N-dimethylformamide (8 mL), and pinacol biborate (902 mg,3.55 mmol) and potassium acetate (435 mg,4.44 mmol) were added. Then under the protection of nitrogen gas, the reaction kettle is provided with a plurality of air channels,pd (dppf) Cl was added 2 (216 mg,296.12 umol). The reaction solution was stirred for 12 hours under nitrogen protection at 80 ℃. The reaction mixture was cooled to room temperature, added to water (20 mL), and extracted with ethyl acetate (10 mL. Times.2). The organic phases were combined and concentrated under reduced pressure to give crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate=100/1 to 10/1) to give the title compound (600 mg, yellow oil), yield: 63.8%. MS (ESI) m/z 318.3[ M+H ]] + .
(3) (R) -2-amino-5- (4- ((3-methylmorpholino) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
(R) -3-methyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) morpholine (80 mg,252.18 mol) was dissolved in a mixed solvent of 1, 4-dioxane (4 mL) and water (0.4 mL), and potassium carbonate (104 mg,756.55 mol) and intermediate 4 (75 mg,252.18 mol) were added. Then Pd (dppf) Cl is added into the reaction solution under the protection of nitrogen 2 (18 mg,25.22 umol). The reaction solution was stirred for 12 hours at 80℃under nitrogen. The reaction solution was cooled and filtered. Purification of the filtrate by Pre-HPLC gave the title compound (36 mg, white solid), yield: 35.1%. MS (ESI) m/z 411.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.43-8.38(m,2H),8.17(d,J＝2.1Hz,1H),7.62(d,J＝8.0Hz,2H),7.38(d,J＝8.0Hz,2H),7.14(s,2H),4.02-3.97(m,2H),3.92-3.88(m,2H),3.66-3.61(m,2H),3.47-3.36(m,3H),3.21-3.13(m,2H),2.54-2.52(m,1H),2.45-2.36(m,1H),2.15-2.07(m,1H),1.79(dd,J＝12.9,1.9Hz,2H),1.62-1.56(m,2H),1.03(d,J＝6.3Hz,3H).
Example 29
Compound 29: 2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) -5- (4- (2-morpholinoethoxy) phenyl) nicotinamide
(1) 4- (2- (4-bromophenoxy) ethyl) morpholine
4- (2-chloroethyl) morpholine (870 mg,5.8 mmol), 4-bromophenol (1.0 g,5.8 mmol) was dissolved in N, N-dimethylformamide (10 mL), potassium carbonate (1.6 g,11.6 mmol) was added thereto, and the temperature was raisedThe reaction was stirred overnight to 70 ℃. Water (20 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (40 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate/petroleum ether=1/3) to give the title compound (1.6 g, colorless oil), yield: 96%. MS (ESI): m/z 285.9[ M+H ]] + .
(2) 2-amino-5- (4- (2-morpholinoethoxy) phenyl) nicotinic acid methyl ester
4- (2- (4-bromophenoxy) ethyl) morpholine (200 mg,0.7 mmol), (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boronic acid (137 mg,0.7 mmol), pdCl 2 (dppf) (57 mg,0.07 mmol), potassium carbonate (193 mg,1.4 mmol) was dissolved in a mixed solvent of 1, 4-dioxane and water (5 mL/1 mL). The reaction was stirred overnight at 85℃under nitrogen. The reaction was filtered, concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (120 mg, yellow oil), yield: 48%. MS (ESI) m/z 358.0[ M+H ] ] + .
(3) 2-amino-5- (4- (2-morpholinoethoxy) phenyl) nicotinic acid
Methyl 2-amino-5- (4- (2-morpholinoethoxy) phenyl) nicotinate (120 mg,0.34 mmol) was dissolved in a mixed solvent of tetrahydrofuran and water (5 mL/1 mL), lithium hydroxide (70 mg,1.68 mmol) was added, and the reaction system was stirred at 40℃overnight. The reaction solution is concentrated to remove tetrahydrofuran, the pH value is regulated to 3-4 by dilute hydrochloric acid, and the solution is concentrated under reduced pressure. The residue was purified by reverse phase column to give the title compound (60 mg, white solid), yield: 52%. MS (ESI) m/z 344.2[ M+H ]] + .
(4) 2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) -5- (4- (2-morpholinoethoxy) phenyl) nicotinamide
2-amino-5- (4- (2-morpholinoethoxy) phenyl) nicotinic acid (60 mg,0.17 mmol) and ((2S, 5R) -5-aminotetrahydro-2H-pyran-2-yl) methanolic hydrochloride (28 mg,0.17 mmol) were dissolved in N, N-dimethylformamide (5 mL), HATU (129 mg,0.34 mmol) and N, N-diisopropylethylamine (66 mg,0.5 mmol) were added and stirred at room temperature overnight. The reaction was purified by Pre-HPLC to give the title compound (45 mg, white solid), yield: 56%. MS (ESI) m/z 457.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.36(d,J＝2.0Hz,1H),8.27(d,J＝7.6Hz,1H),8.10(d,J＝2.0Hz,1H),7.58(d,J＝8.8Hz,2H),7.06-7.01(m,4H),4.63(t,J＝5.6Hz,1H),4.12(t,J＝5.6Hz,2H),3.91-3.89(m,2H),3.60-3.57(m,4H),3.40-3.35(m,1H),3.34-3.31(m,1H),3.13(t,J＝11.8Hz,1H),2.70(t,J＝6.0Hz,2H),2.47-2.45(m,4H),1.97(d,J＝12.4Hz,1H),1.73(d,J＝14.0Hz,1H),1.65-1.54(m,1H),1.36-1.26(m,1H).
Example 30
Compound 30:5- (4- (2-oxa-6-azaspiro [3.3] heptan-6-yl) phenyl) -2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) nicotinamide
(1) 6- (4-bromophenyl) -2-oxa-6-azaspiro [3.3] heptane
1, 4-Dibromobenzene (800 mg,3.39 mmol), tris (dibenzylideneacetone) dipalladium (30 mg,0.033 mmol), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (42 mg,0.067 mmol), 2-oxo-6-azaspiro [3.3] in a 25mL single-port flask]Heptane (224 mg,2.26 mmol) and cesium carbonate (1.1 g,3.37 mmol) were added toluene (5 mL) and reacted at 100 ℃ for 18 hours under nitrogen protection. The reaction was concentrated, and the residue was purified by column chromatography (petroleum ether/ethyl acetate=2/1) to give the title compound (190 mg, white solid), yield: 38%. MS (ESI) M/z253.9[ M+H ]] + .
(2) 5- (4- (2-oxa-6-azaspiro [3.3] heptan-6-yl) phenyl) -2-aminonicotinic acid methyl ester
6- (4-bromophenyl) -2-oxa-6-azaspiro [3.3] was sequentially added to a 50mL single vial]Heptane (158 mg,0.62 mmol), (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boronic acid (134 mg,0.68 mmol), pd (dppf) Cl 2 (45 mg,0.062 mmol) and potassium carbonate (258 mg,1.87 mmol) were added 1, 4-dioxane (4 mL) and water (2 mL) and reacted at 80℃for 16 hours under nitrogen protection. The solvent was removed under reduced pressure and the residue was purified by column chromatography (dichloromethane/methanol=20/1) to give the title compound (17 mg, yellow solid), yield: 8.4%. MS (ESI) M/z326.0[ M+H ] ] + .
(3) 5- (4- (2-oxa-6-azaspiro [3.3] heptan-6-yl) phenyl) -2-amino nicotinic acid
5- (4- (2-oxa-6-azaspiro [ 3.3)]Heptane-6-yl) phenyl) -2-amino methyl nicotinate (17 mg,0.052 mmol) was dissolved in methanol (3 mL) and water (1 mL), sodium hydroxide (6.24 mg,0.156 mmol) was added, and the reaction was heated to 50 ℃ for 4 hours. The reaction was dried by spin and the residue was purified by reverse phase column to give the title compound (13 mg, yellow solid), yield: 81%. MS (ESI) m/z 311.9[ M+H ]] + .
(4) 5- (4- (2-oxa-6-azaspiro [3.3] heptan-6-yl) phenyl) -2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) nicotinamide
5- (4- (2-oxa-6-azaspiro [ 3.3)]Heptan-6-yl) phenyl) -2-aminonicotinic acid (13 mg,0.042 mmol) was dissolved in N, N-dimethylformamide (2 mL), ((2S, 5R) -5-aminotetrahydro-2H-pyran-2-yl) methanolic hydrochloride (8.4 mg,0.05 mmol), HATU (32 mg,0.084 mmol) and N, N-diisopropylethylamine (27 mg,0.21 mmol) were added and the reaction stirred at room temperature for an additional 3 hours. The reaction was filtered and the filtrate purified by Pre-HPLC to give the title compound (13 mg, pale yellow solid), yield: 48%. MS (ESI) m/z 425.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.31(d,J＝2.4Hz,1H),8.25(d,J＝7.6Hz,1H),8.05(d,J＝2.0Hz,1H),7.47(d,J＝8.4Hz,2H),6.99(s,2H),6.51(d,J＝8.4Hz,2H),4.73(s,4H),4.64(t,J＝5.6Hz,1H),3.98(s,4H),3.92-3.85(m,2H),3.43-3.37(m,2H),3.28-3.22(m,1H),3.13(t,J＝11.6Hz,1H),1.98-1.95(m,1H),1.75-1.71(m,1H),1.64-1.55(m,1H),1.36-1.25(m,1H).
Example 31
Compound 31:5- (4- (1- (2-oxa-6-azaspiro [3.3] heptan-6-yl) ethyl) phenyl) -2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) nicotinamide
(1) 6- (1- (4-bromophenyl) ethyl) -2-oxo-6-azaspiro [3.3] heptane
1- (4-bromophenyl) ethanone (500 mg,2.5 mmol) was dissolved in dichloromethane (5 mL) and 2-oxa-6-azaspiro [3.3] was added]Heptane (2793 mg, 2)72 mmol) was stirred at room temperature for 2 hours, sodium cyanoborohydride (473 mg,7.5 mmol) was added and the reaction was allowed to stand overnight at room temperature. The reaction was purified by column chromatography to give the title compound (200 mg, yellow oil), yield: 28%. MS (ESI) m/z 281.9[ M+H ]] + .
(2) 5- (4- (1- (2-oxa-6-azaspiro [3.3] heptan-6-yl) ethyl) phenyl) -2-amino nicotinic acid methyl ester
6- (1- (4-bromophenyl) ethyl) -2-oxo-6-azaspiro [3.3]Heptane (200 mg,0.71 mmol) was dissolved in 1, 4-dioxane (2 mL) and water (2 mL), and (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boronic acid (138 mg,0.71 mmol), potassium carbonate (292 mg,2.13 mmol) and Pd (dppf) Cl were added sequentially 2 (50 mg,0.071 mmol). The reaction solution was reacted for 2.5 hours under the protection of nitrogen at 70 ℃. Filtration, concentration of the filtrate, and purification of the residue by column chromatography gave the title compound (60 mg, yellow oil), yield: 24%. MS (ESI) m/z 354.1[ M+H ]] + .
(3) 5- (4- (1- (2-oxa-6-azaspiro [3.3] heptan-6-yl) ethyl) phenyl) -2-amino nicotinic acid
5- (4- (1- (2-oxa-6-azaspiro [3.3 ]) ]Heptane-6-yl) ethyl) phenyl) -2-amino methyl nicotinate (60 mg,0.17 mmol) was dissolved in a mixed solvent of tetrahydrofuran (1 mL) and water (1 mL), and sodium hydroxide (20 mg,0.54 mmol) was added. The reaction was stirred at room temperature for 4 hours. The reaction was purified by reverse phase column to give the title compound (20 mg, grey solid), yield: 35%. MS (ESI) m/z 340.1[ M+H ]] + .
(4) 5- (4- (1- (2-oxa-6-azaspiro [3.3] heptan-6-yl) ethyl) phenyl) -2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) nicotinamide
5- (4- (1- (2-oxa-6-azaspiro [3.3 ])]Heptane-6-yl) ethyl) phenyl) -2-aminonicotinic acid (10 mg,0.03 mmol) was dissolved in N, N-dimethylformamide (1 mL), and ((2 s,5 r) -5-aminotetrahydro-2H-pyran-2-yl) methanolic hydrochloride (7.6 mg,0.06 mmol), HATU (16 mg,0.045 mmol) and N, N-diisopropylethylamine (7.3 mg,0.06 mmol) were added sequentially. The reaction solution was stirred at room temperature for 16 hours. The reaction was purified by Pre-HPLC to give the title compound (2 mg, white solid), yield: 15%. MS (ESI): m/z 453.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.36(s,1H),8.26(d,J＝7.6Hz,1H),8.14(s,1H),7.57(d,J＝8.0Hz,2H),7.33(d,J＝7.2Hz,2H),7.13(s,2H),4.64-4.59(m,5H),3.91-3.89(m,3H),3.16-3.09(m,5H),2.01-1.96(m,2H),1.77-1.54(m,3H),1.37-1.24(m,2H),1.10(d,J＝6.0Hz,3H).
Example 32
Compound 32: 2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) -5- (4- (1- (pyrrolidin-1-yl) ethyl) phenyl) nicotinamide
(1) 2-amino-5-bromo-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) nicotinamide
The procedure of example 10, step (1), was followed using 2-amino-5-bromonicotinic acid (1.61 g,7.43 mmol) and ((2 s,5 r) -5-aminotetrahydro-2H-pyran-2-yl) methanol (650 mg,4.96 mmol) as raw materials to give the title compound (300 mg, white solid), yield: 18.34%. MS (ESI) m/z 330.1[ M+H ]] + . 1 H NMR(400MHz,CD 3 OD)δ8.11(d,J＝2.4Hz,1H),8.02(d,J＝2.4Hz,1H),4.09-3.98(m,2H),3.53(d,J＝5.2Hz,2H),3.44-3.38(m,1H),3.26-3.20(m,1H),2.12-2.08(m,1H),1.77(dd,J＝12.9,2.5Hz,1H),1.68-1.60(m,1H),1.53-1.41(m,1H).
(2) 2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) -5- (4- (1- (pyrrolidin-1-yl) ethyl) phenyl) nicotinamide
2-amino-5-bromo-N- ((3R, 6S) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) nicotinamide (100 mg, 302.87. Mu. Mol) was dissolved in a mixed solvent of 1, 4-dioxane (0.5 mL) and water (0.5 mL), 1- (1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) ethyl) pyrrolidine (136 mg, 454.30. Mu. Mol, example 25, step 2) and potassium carbonate (125 mg, 908.60. Mu. Mol) were added sequentially, pd (dppf) Cl under nitrogen protection 2 (44 mg,60.57 umol). The reaction solution was stirred at 80℃for 12 hours. The reaction solution was cooled to room temperature, diluted with water, and extracted with ethyl acetate (5 mL). The organic phase was concentrated under reduced pressure to give crude product. Purification of the crude product by Prep-HPLC gave the title compound (4 mg, grey solid), yield: 3.2%. MS (ESI) m/z 425.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.39(d,J＝2.1Hz,1H),8.28(d,J＝7.4Hz,1H),8.15(d,J＝2.3Hz,1H),7.58(d,J＝8.2Hz,2H),7.38(d,J＝8.2Hz,2H),7.14(s,2H),4.65(t,J＝5.8Hz,1H),3.95-3.83(m,2H),3.44-3.37(m,1H),3.36-3.33(m,1H),3.32-3.28(m,2H),3.27-3.19(m,2H),3.17-3.10(m,1H),2.33(m,2H),2.03-1.93(m,1H),1.77-1.56(m,6H),1.31(d,J＝6.6Hz,3H),1.30-1.24(m,1H).
Example 33
Compound 33: 2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) -5- (4- (morpholinomethyl) phenyl) nicotinamide
Referring to the procedure of example 10 step (2), starting from 2-amino-5-bromo-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) nicotinamide (108 mg,329umol, example 32, step 1) and 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) morpholine (100 mg, 399 umol), the title compound (71.2 mg, white solid) was obtained in the yield: 50.62%. MS (ESI) m/z 427.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.41(d,J＝2.3Hz,1H),8.29(d,J＝7.5Hz,1H),8.16(d,J＝2.3Hz,1H),7.62(d,J＝8.0Hz,2H),7.37(d,J＝8.0Hz,2H),7.15(s,2H),4.65(t,J＝5.8Hz,1H),3.95-3.85(m,2H),3.58(t,J＝4.5Hz,4H),3.49(s,2H),3.44-3.37(m,1H),3.36-3.33(m,1H),3.30-3.22(m,1H),3.18-3.10(m,1H),2.40-2.32(m,4H),1.98(d,J＝12.2Hz,1H),1.74(d,J＝13.9Hz,1H),1.61-1.57(m,1H),1.37-1.23(m,1H).
Example 34
Compound 34: 3-amino-6- (4- (morpholinomethyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) pyrazine-2-carboxamide
(1) 3-amino-6-bromo-N- (tetrahydro-2H-pyran-4-yl) pyrazine-2-carboxamide
tetrahydro-2H-pyranes4-amine (300 mg,2.97 mmol) and 3-amino-6-bromopyrazine-2-carboxylic acid (500 mg,2.29 mmol) were dissolved in dichloromethane (5 mL), triethylamine (465.28 mg,4.60 mmol) and tri-n-propyl cyclic phosphoric anhydride (2.19 g,3.44mmol,50% ethyl acetate solution) were added, and the reaction was stirred at 15℃for 16 hours. The reaction was poured into water (10 mL) and the aqueous phase was extracted with ethyl acetate (5 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purification of the crude product by Prep-TLC (petroleum ether/ethyl acetate=1/1) gave the title compound (200 mg, off-white solid), yield: 28.96%. MS (ESI) m/z 301.1[ M+H ] ] + . 1 H NMR(400MHz,CDCl 3 )δ8.15(s,1H),7.60-7.37(m,1H),4.10-3.89(m,3H),3.50-3.42(m,2H),1.93-1.85(m,2H),1.64-1.55(m,2H).
(2) 3-amino-6- (4- (morpholinomethyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) pyrazine-2-carboxamide
Reference to the procedure of example 10 step (2) starting from 3-amino-6-bromo-N- (tetrahydro-2H-pyran-4-yl) pyrazine-2-carboxamide (70 mg,232.45 mol) and 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) morpholine (90 mg,296.83 mol) gave the title compound (34.6 mg, pale yellow solid), yield: 37.45%. MS (ESI) m/z 398.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.82(s,1H),8.52(d,J＝8.4Hz,1H),8.08(d,J＝8.0Hz,2H),7.62(brs,2H),7.41(d,J＝8.0Hz,2H),4.11-3.99(m,1H),3.96-3.87(m,2H),3.63-3.56(m,4H),3.52(s,2H),3.46-3.38(m,2H),2.39-2.37(m,4H),1.84-1.71(m,4H).
Example 35
Compound 35: 2-amino-N- ((3 r,6 s) -6- (2-hydroxy-propan-2-yl) tetrahydro-2H-pyran-3-yl) -5- (4- (morpholinomethyl) phenyl) nicotinamide
Reference to example 10 procedure (2) was followed as 2-amino-5-bromo-N- ((3R, 6S) -6- (2-hydroxypropan-2-yl) tetrahydro-2H-pyran-3-yl) nicotinamide (35.4 mg, 98.9. Mu. Mol, example 10, step 1) and 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl)The title compound (19.2 mg, yellow solid) was obtained in the yield of: 42.69%. MS (ESI) m/z 455.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.41(d,J＝2.3Hz,1H),8.28(d,J＝7.6Hz,1H),8.16(d,J＝2.1Hz,1H),7.62(d,J＝8.1Hz,2H),7.38(d,J＝8.1Hz,2H),7.15(s,2H),4.25(s,1H),3.97-3.92(m,1H),3.91-3.84(m,1H),3.58(t,J＝4.4Hz,4H),3.49(s,2H),3.13(t,J＝10.3Hz,1H),2.98(d,J＝11.2Hz,1H),2.37-2.35(m,4H),2.00(d,J＝12.4Hz,1H),1.82(d,J＝13.0Hz,1H),1.60-1.55(m,1H),1.45-1.33(m,1H),1.10(s,3H),1.05(s,3H).
Example 36
Compound 36: 2-amino-5- (4- (morpholinomethyl) phenyl) -N- (2-oxaspiro [3.3] heptane-6-yl) nicotinamide
(1) 2-amino-5- (4- (morpholinomethyl) phenyl) nicotinic acid methyl ester
Methyl 2-amino-5-bromonicotinate (175.85 mg,761.10 umol) and 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) morpholine (300 mg,989.44 umol) were dissolved in 1, 4-dioxane (3 mL) and water (0.6 mL). Sodium carbonate (242.01 mg,2.28 mmol) and Pd (dppf) Cl were added to the reaction solution 2 (55.69 mg,76.11 umol). The reaction solution was replaced with nitrogen three times, heated to 90℃and stirred for 16 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated. Purification of the crude product by Prep-TLC (ethyl acetate/methanol=10/1) gave the title compound (180 mg, yellow solid), yield: 72.24%. MS (ESI) m/z 328.2[ M+H ]] + . 1 H NMR(400MHz,CDCl 3 )δ8.42(d,J＝2.4Hz,1H),8.28(d,J＝2.4Hz,1H),7.40(d,J＝8.0Hz,2H),7.32(d,J＝8.0Hz,2H),6.38(brs,2H),3.85(s,3H),3.65(t,J＝4.4Hz,4H),3.46(s,2H),2.40(t,J＝4.4Hz,4H).
(2) 2-amino-5- (4- (morpholinomethyl) phenyl) nicotinic acid
Methyl 2-amino-5- (4- (morpholinomethyl) phenyl) nicotinate (100 mg,305.46 mol) was dissolved in methanol (1 mL) and water (1 mL), and lithium hydroxide monohydrate (30) was added to the reaction solutionmg,714.91 umol). The reaction solution was stirred at 15℃for 16 hours. The reaction mixture was concentrated to remove methanol, and water (2 mL) and ethyl acetate (2 mL) were added for liquid extraction. The pH value of the water phase is regulated to 2-3 by 1M dilute hydrochloric acid, and then the water phase is directly freeze-dried. The crude product was dispersed in methanol (5 mL) and stirred for 2 hours, then filtered, and the filtrate was concentrated to give the title compound (100 mg, yellow solid), the crude product was used directly in the next step. MS (ESI) m/z 314.2[ M+H ] ] + .
(3) 2-amino-5- (4- (morpholinomethyl) phenyl) -N- (2-oxaspiro [3.3] heptane-6-yl) nicotinamide
2-amino-5- (4- (morpholinomethyl) phenyl) nicotinic acid (30 mg, 95.74. Mu. Mol) and 2-oxaspiro [3.3]Heptan-6-amine hydrochloride (15 mg,100.26 umol), HATU (40.04 mg,105.31 umol) was dissolved in N, N-dimethylformamide (0.5 mL), and N, N-diisopropylethylamine (37.10 mg,287.06 umol) was added. The reaction solution was stirred at 15℃for 3 hours. The reaction was purified by Prep-HPLC to give the title compound (14.8 mg, white solid), yield: 37.84%. MS (ESI) m/z 409.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.64(d,J＝7.0Hz,1H),8.41(d,J＝2.1Hz,1H),8.16(d,J＝2.1Hz,1H),7.62(d,J＝8.0Hz,2H),7.38(d,J＝8.0Hz,2H),7.15(s,2H),4.64(s,2H),4.52(s,2H),4.30-4.17(m,1H),3.58(t,J＝4.5Hz,4H),3.49(s,2H),2.62-2.56(m,2H),2.37-2.35(m,4H),2.29-2.21(m,2H).
Example 37
Compound 37: 2-amino-5- (3-fluoro-4- (morpholinomethyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
Referring to the procedure of example 26, starting from 4- (2-fluoro-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) morpholine (50 mg,155.67 mol) and intermediate 4 (46 mg,155.67 mol), the title compound (12 mg, white solid) was obtained in the yield: 18.9%. MS (ESI) m/z 415.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.47(d,J＝2.3Hz,1H),8.40(d,J＝7.7Hz,1H),8.19(d,J＝2.3Hz,1H),7.55(d,J＝11.7Hz,1H),7.53-7.44(m,2H),7.24(s,2H),4.08-3.95(m,1H),3.93-3.87(m,2H),3.60-3.56(m,4H),3.55(s,2H),3.40(t,J＝11.0Hz,2H),2.41-2.39(m,4H),1.81-1.77(m,2H),1.64-1.53(m,2H).
Example 38
Compound 38: 2-amino-5- (4- ((4-hydroxypiperidin-1-yl) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
(1) 1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) piperidin-4-ol
2- (4- (bromomethyl) phenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborolan (200 mg,673.41 mmol) was dissolved in acetonitrile (2 mL), triethylamine (136 mg,1.35 mmol) and piperidin-4-ol (136 mg,1.35 mmol) were added, and the reaction solution was stirred at 20℃for 3 hours. The reaction solution was concentrated under reduced pressure to give the title compound (0.2 g, brown solid), and the crude product was used in the next step without purification. MS (ESI) m/z 318.3[ M+H ]] + .
(2) 2-amino-5- (4- ((4-hydroxypiperidin-1-yl) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
Referring to the procedure of example 26, starting from 1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) piperidin-4-ol (50 mg,157.61 mol) and intermediate 4 (47 mg,157.61 mol), the title compound (12 mg, white solid) was obtained in yield: 19.0%. MS (ESI) m/z 411.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.44-8.39(m,2H),8.17(d,J＝2.2Hz,1H),7.61(d,J＝8.1Hz,2H),7.35(d,J＝8.1Hz,2H),7.15(s,2H),4.55(d,J＝3.4Hz,1H),4.08-3.96(m,1H),3.90(dd,J＝11.2,2.2Hz,2H),3.46(s,2H),3.44-3.36(m,3H),2.72-2.62(m,2H),2.03(t,J＝9.8Hz,2H),1.79(dd,J＝12.5,2.1Hz,2H),1.74-1.67(m,2H),1.60-1.56(m,2H),1.44-1.33(m,2H).
Example 39
Compound 39: (2-amino-5- (4- (morpholinomethyl) phenyl) pyridin-3-yl) (4-hydroxypiperidin-1-yl) methanone
Referring to the procedure of example 36, step (3), starting from 2-amino-5- (4- (morpholinomethyl) phenyl) nicotinic acid (30 mg,95.85umol, example 36, step 2) and piperidin-4-ol (11 mg,105.43 umol), the title compound (14.2 mg, yellow solid) was obtained in yield: 37.4%. MS (ESI) m/z 397.3[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.35(d,J＝2.3Hz,1H),7.65(d,J＝2.1Hz,1H),7.56(d,J＝8.0Hz,2H),7.34(d,J＝8.0Hz,2H),6.05(s,2H),4.77(d,J＝3.3Hz,1H),4.01-3.62(m,3H),3.58(t,J＝4.3Hz,4H),3.47(s,2H),3.19(t,J＝9.4Hz,2H),2.37-2.35(m,4H),1.76-1.74(m,2H),1.41-1.39(m,2H).
Example 40
Compound 40: (2-amino-5- (4- (morpholinomethyl) phenyl) pyridin-3-yl) (4- (hydroxymethyl) piperidin-1-yl) methanone
Referring to the procedure of example 36, step (3), starting from 2-amino-5- (4- (morpholinomethyl) phenyl) nicotinic acid (30 mg,95.85umol, example 36, step 2) and piperidin-4-ylmethanol (12 mg,105.43 umol), the title compound (16.9 mg, yellow oil) was obtained in yield: 43.0%. MS (ESI) m/z 411.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.34(d,J＝2.1Hz,1H),7.63(d,J＝2.1Hz,1H),7.56(d,J＝8.0Hz,2H),7.35(d,J＝8.0Hz,2H),6.04(s,2H),4.49(brs,1H),3.58(t,J＝4.4Hz,4H),3.47(s,2H),3.28-3.26(m,4H),2.92-2.89(m,2H),2.37-2.35(m,4H),1.76-1.57(m,3H),1.15-1.11(m,2H).
Example 41
Compound 41: 2-amino-N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) -5- (1- (1- (oxa-3-yl) piperidin-4-yl) -1H-indazol-5-yl) nicotinamide
(1) 4- (5-bromo-1H-indazol-1-yl) piperidine-1-carboxylic acid tert-butyl ester
1- (t-Butoxycarbonyl) -4- (methylsulfonyloxy) piperidine (4.68 g,16.75 mmol) was added in portions to a solution of 5-bromoindazole (3 g,15.23 mmol) and sodium hydride (670.06 mg,16.75mmol,60% purity) in N, N-dimethylformamide (30 mL) at 0deg.C. The reaction solution was warmed to 100℃and stirred under nitrogen for 14 hours. The reaction was poured into saturated ammonium chloride (50 mL) and quenched, extracted with ethyl acetate (10 mL x 3) and the organic phase concentrated to give crude product. Purification of the crude product by column chromatography (petroleum ether/ethyl acetate=3/1-1/1) gave tert-butyl 4- (5-bromo-1H-indazol-1-yl) piperidine-1-carboxylate (2.8 g, white solid), yield: 48.35%. 1 H NMR(400MHz,CDCl 3 )δ7.86(s,1H),7.80(d,J＝1.5Hz,1H),7.37(dd,J＝8.9,1.6Hz,1H),7.26(d,J＝8.9Hz,1H),4.49-4.42(m,1H),4.25-4.22(m,2H),2.92-2.85(m,2H),2.20-2.07(m,2H),1.96-1.87(m,2H),1.42(s,9H).
Meanwhile, the reaction was separated and purified to give another isomer, tert-butyl 4- (5-bromo-2H-indazol-2-yl) piperidine-1-carboxylate (1.2 g, white solid), yield: 20.72%. 1 H NMR(400MHz,CDCl 3 )δ7.83(s,1H),7.74(d,J＝1.3Hz,1H),7.52(d,J＝9.1Hz,1H),7.28(dd,J＝9.1,1.8Hz,1H),4.53-4.45(m,1H),4.31-4.19(m,2H),2.94-2.81(m,2H),2.17(dd,J＝12.1,1.8Hz,2H),2.09-1.95(m,2H),1.42(s,9H).
(2) 5-bromo-1- (piperidin-4-yl) -1H-indazole
Tert-butyl 4- (5-bromo-1H-indazol-1-yl) piperidine-1-carboxylate (2 g,5.26 mmol) was dissolved in dichloromethane (8 mL) and a solution of hydrochloric acid/dioxane (4M, 25 mL) was added. The reaction solution was stirred at 15℃for 12 hours. The reaction was concentrated at 45℃to give the title compound (1.67 g, hydrochloride salt, white solid), the crude product was used directly in the next step. MS (ESI) m/z 280.0[ M+H ]] + .
(3) 5-bromo-1- (1- (oxetan-3-yl) piperidin-4-yl) -1H-indazole
5-bromo-1- (piperidin-4-yl) -1H-indazole (300 mg,947.50umol, hydrochloride) was dissolved in methanol (3 mL) at 15℃and triethylamine (95.88 mg,947.50 umol) and 3-oxetanone (94.15 mg,1.31 mmol) were added. After the reaction solution was stirred at 15℃for 1 hour, cyanoboron was added to the reaction solutionSodium hydride (120.0 mg,1.91 mmol) and acetic acid (63.0 mg,1.05 mmol). The reaction mixture was stirred at 15℃for a further 16 hours. Water (5 mL) was added to the reaction solution, extraction was performed with ethyl acetate (5 mL. Times.4), and the organic phases were combined and concentrated to give the crude product. Purification of the crude product by Prep-TLC (dichloromethane/methanol=10/1) gave the title compound (170 mg, white solid), yield: 53.36%. MS (ESI): M/z336.1[ M+H ] ] + . 1 H NMR(400MHz,CDCl 3 )δ7.86(s,1H),7.80(s,1H),7.39-7.35(m,1H),7.31-7.27(m,1H),4.65-4.58(m,4H),4.41-4.30(m,1H),3.56-3.49(m,1H),2.88(d,J＝12.5Hz,2H),2.40-2.26(m,2H),2.10-1.94(m,4H).
(4) 1- (1- (oxetan-3-yl) piperidin-4-yl) -5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indazole
Dipinacol diboronate (79.30 mg,312.29 mol) and 5-bromo-1- (1- (oxetan-3-yl) piperidin-4-yl) -1H-indazole (70 mg,208.19 mol) were dissolved in 1, 4-dioxane (1 mL) at 15℃and potassium phosphate (132.58 mg,624.58 mol) and 1, 1-bis (diphenylphosphino) ferrocene palladium (II) dichloride dichloromethane complex (17.00 mg,20.82 mol) were added. The reaction solution was purged with nitrogen and then heated to 80℃and stirred for 16 hours. After the reaction mixture was cooled to room temperature, methanol (5 mL) was added thereto for dilution, and the mixture was filtered. The filtrate was purified by Prep-TLC (ethyl acetate/methanol=10/1) to give the title compound (45 mg, white solid), yield: 56.39%. MS (ESI) m/z 384.3[ M+H ]] + . 1 H NMR(400MHz,CDCl 3 )δ8.20(s,1H),7.93(s,1H),7.70(d,J＝8.2Hz,1H),7.38(d,J＝8.6Hz,1H),4.68-4.55(m,4H),4.39-4.34(m,1H),3.54-3.49(m,1H),2.91-2.81(m,2H),2.42-2.28(m,2H),2.07-1.94(m,4H),1.30(s,12H).
(5) 2-amino-N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) -5- (1- (1- (oxa-3-yl) piperidin-4-yl) -1H-indazol-5-yl) nicotinamide
Intermediate 5 (33.29 mg,97.84 mol) and 1- (1- (oxetan-3-yl) piperidin-4-yl) -5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indazole (45 mg,117.40 mol) were dissolved in a mixed solvent of 1, 4-dioxane (0.5 mL) and water (0.1 mL) at 15℃and sodium carbonate (31.11 mg,293.51 mol) and Pd (dppf) Cl were added 2 (7.16 mg,9.78 umol). Reaction liquidThe mixture was aerated with nitrogen and then heated to 80℃and stirred for 16 hours. The reaction was cooled to room temperature and concentrated, the crude product was diluted with methanol (3 mL) and filtered, and the filtrate was purified by Prep-HPLC to give the title compound (5.5 mg, white solid), yield: 10.66%. MS (ESI) m/z 517.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.39(d,J＝2.4Hz,1H),8.12(s,1H),8.04(d,J＝2.3Hz,1H),7.99(s,1H),7.81-7.78(m,2H),7.69(dd,J＝8.8,1.6Hz,1H),6.88(s,2H),4.72-4.63(m,1H),4.61-4.56(m,2H),4.48(t,J＝6.1Hz,2H),4.31(s,1H),3.54-3.44(m,1H),2.86(d,J＝10.9Hz,2H),2.22-2.11(m,2H),2.10-2.03(m,8H),1.95(d,J＝11.8Hz,2H),1.67-1.59(m,6H).
Example 42
Compound 42: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (4- (oxa-3-yl) piperazin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinamide
(1) 1- (5-bromo-2, 3-dihydro-1H-inden-1-yl) -4- (oxetan-3-yl) piperazine
5-bromo-2, 3-dihydro-1H-inden-1-one (3.00 g,14.21 mmol) was dissolved in tetrahydrofuran (150 mL), 1- (oxetan-3-yl) piperazine (4.04 g,28.43 mmol) was added sequentially, tetraisopropyl titanate (12.12 g,42.64 mmol) was added, and the reaction was stirred at 70℃for 4 hours. Then, the reaction was cooled to 20℃and sodium triacetoxyborohydride (12.05 g,56.86 mmol) was added to the reaction solution. The reaction solution was stirred at 20℃for 12 hours. The reaction mixture was added to water (200 mL), and a white solid was precipitated and filtered. The filtrate was extracted with ethyl acetate (100 ml x 2) and the combined organic phases concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=100/1 to 0/1) to give the title compound (700 mg, black oil), yield: 14.6%. MS (ESI) m/z 337.1[ M+H ] ] + .
(2) 1- (oxetan-3-yl) -4- (5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -2, 3-dihydro-1H-inden-1-yl) piperazine
1- (5-bromo-2, 3-dihydro-1H-inden-1-yl) -4- (oxetan-3-yl) piperazine (350 mg)1.04 mmol) was dissolved in 1, 4-dioxane (3.5 mL) and potassium acetate (305 mg,3.11 mmol) was added, pinacol biborate (316 mg,1.25 mmol). Pd (dppf) Cl was added under nitrogen blanket 2 (75 mg,103.78 umol). The reaction solution was stirred for 12 hours at 80℃under nitrogen. The reaction solution was cooled to room temperature, water (15 mL) and ethyl acetate (15 mL) were added and the mixture was extracted in portions, and the organic phase was concentrated under reduced pressure to give a crude product. Purification of the crude product by Prep-TLC (ethyl acetate/methanol=0/1) gave the title compound (200 mg, yellow solid), yield: 50.1%. MS (ESI) m/z 385.3[ M+H ]] + .
(3) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (4- (oxa-3-yl) piperazin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinamide
1- (oxetan-3-yl) -4- (5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -2, 3-dihydro-1H-inden-1-yl) piperazine (50 mg,130.10 umol) was dissolved in a mixed solution of 1, 4-dioxane (0.5 mL) and water (0.5 mL), and intermediate 5 (44 mg,130.10 umol) and potassium carbonate (53 mg,390.30 umol) were added. Pd (dppf) Cl was added to the reaction solution under nitrogen protection 2 (9 mg,13.01 umol). The reaction solution was stirred for 12 hours at 80℃under nitrogen. The reaction solution was cooled to room temperature, water (5 mL) and ethyl acetate (5 mL) were added and the mixture was extracted in portions, and the organic phase was concentrated under reduced pressure to give a crude product. Purification of the crude product by Prep-HPLC gave (5 mg, white solid), yield: 7.4%. MS (ESI) m/z 518.4[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.33(d,J＝2.3Hz,1H),8.00(d,J＝2.1Hz,1H),7.77(s,1H),7.49(s,1H),7.45(d,J＝7.7Hz,1H),7.31(d,J＝7.9Hz,1H),6.91(s,2H),4.55-4.49(m,2H),4.43-4.37(m,2H),4.31-4.29(m,2H),3.42-3.37(m,1H),2.99-2.88(m,1H),2.86-2.77(m,1H),2.55-2.52(m,2H),2.46-2.39(m,2H),2.35-2.14(m,4H),2.08-2.00(m,8H),1.65-1.59(m,6H).
Example 43
Compound 43: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinamide
(1) 3- (5-bromo-1H-indazol-1-yl) pyrrolidine-1-carboxylic acid tert-butyl ester
5-bromo-1H-indazole (3.00 g,15.2 mmol) was dissolved in N, N-dimethylformamide (30 mL), cooled to 0deg.C, sodium hydride (669 mg,16.7mmol,60% purity) was slowly added, and stirred at 0deg.C for 30 min. Then 3- ((methylsulfonyl) oxy) pyrrolidine-1-carboxylic acid tert-butyl ester (4.44 g,16.7 mmol) was added to the reaction solution at 0 ℃. The reaction solution was heated to 100℃and stirred for 14 hours. The reaction mixture was cooled to room temperature, water (100 mL) was added to the reaction mixture, and the mixture was extracted with methylene chloride (50 mL. Times.2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate=5/1 to 1/1) to give 3- (5-bromo-1H-indazol-1-yl) pyrrolidine-1-carboxylic acid tert-butyl ester (2.90 g, white solid), yield: 52.0%. MS (ESI) m/z 310.0[ M+H-56 ] ] + . 1 H NMR(400MHz,CDCl 3 )δ7.87(s,1H),7.81(s,1H),7.40(d,J＝8.7Hz,1H),7.25(d,J＝8.7Hz,1H),5.11-5.01(m,1H),3.92-3.46(m,4H),2.56-2.28(m,2H),1.40(s,9H).
Meanwhile, the reaction was separated and purified to give another isomer, tert-butyl 3- (5-bromo-2H-indazol-2-yl) pyrrolidine-1-carboxylate (2.0 g, white solid), yield: 35.86%. MS (ESI) m/z 310.0[ M+H-56 ]] + . 1 H NMR(400MHz,CDCl 3 )δ7.83(s,1H),7.73(d,J＝1.1Hz,1H),7.51(d,J＝9.2Hz,1H),7.27(dd,J＝9.2,1.5Hz,1H),5.10-5.07(m,1H),3.96-3.72(m,2H),3.70-3.48(m,2H),2.48-2.40(m,2H),1.41(s,9H).
(2) 5-bromo-1- (pyrrolidin-3-yl) -1H-indazole
3- (5-bromo-1H-indazol-1-yl) pyrrolidine-1-carboxylic acid tert-butyl ester (500 mg,1.37 mmol) was dissolved in dichloromethane (5.0 mL) and a hydrochloric acid/dioxane solution (4.0M, 1.02 mL) was slowly added dropwise. The reaction solution was stirred at room temperature for 12 hours. The reaction solution was concentrated to give the title compound (300 mg, white solid, hydrochloride). MS (ESI) m/z 266.0[ M+H ]] + . 1 H NMR(400MHz,CD 3 OD)δ8.10(s,1H),7.99(d,J＝1.2Hz,1H),7.66-7.63(m,1H),7.59-7.55(m,1H),5.71-5.63(m,1H),3.91-3.76(m,2H),3.73-3.55(m,2H),2.64-2.58(m,1H),2.41-2.31(m,1H).
(3) 5-bromo-1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazole
5-bromo-1- (pyrrolidin-3-yl) -1H-indazole (300 mg,991umol, hydrochloride) was dissolved in methanol (3.0 mL), triethylamine (100 mg,991 umol) and tetrahydro-4H-pyran-4-one (138 mg,1.39 mmol) were added, and the reaction was stirred at 15℃for 1 hour. To the reaction solution were added sodium cyanoborohydride (124 mg,1.98 mmol) and acetic acid (65.4 mg,1.09 mmol), and stirring was continued for 12 hours. To the reaction mixture was added water (10 mL), followed by extraction with ethyl acetate (5 mL. Times.2). The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give the title compound (110 mg, yellow oil). MS (ESI) m/z 350.1[ M+H ] ] + . 1 H NMR(400MHz,CDCl 3 )δ7.87(s,1H),7.80(s,1H),7.40(s,1H),7.19(s,1H),5.28-5.26(m,1H),3.98-3.94(m,2H),3.56-3.53(m,1H),3.37-3.31(m,2H),3.21-2.93(m,3H),2.72-2.44(m,2H),2.32-2.22(m,1H),1.87-1.66(m,4H).
(4) 2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinic acid methyl ester
5-bromo-1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazole (110 mg,314 umol) and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (104 mg,376 umol) were dissolved in 1, 4-dioxahexacyclic (1.1 mL) and water (0.22 mL), and potassium carbonate (86.8 mg, 238 umol) was added. The system was replaced three times with nitrogen and Pd (dppf) Cl was added under nitrogen blanket 2 (22.9 mg,31.4 umol). The reaction solution was stirred at 100℃for 12 hours. After the reaction mixture was cooled to room temperature, water (5 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 mL. Times.2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purification of the crude product by Prep-TLC gave the title compound (90.0 mg, yellow oil), yield: 67.8%. MS (ESI) m/z 422.3[ M+H ]] + .
(5) 2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinate (90.0 mg,213 umol) was dissolved in methanol (0.9 mL) and water (0.9 mL) and lithium hydroxide (10.2 mg,427 umol) was added. The reaction solution was stirred at room temperature for 12 hours. The reaction liquid is concentrated to obtain a concentrated solution, Purification of the crude product by Prep-HPLC gave the title compound (30.0 mg, colorless oil), yield: 34.48%. MS (ESI) m/z 408.2[ M+H ]] + .
(6) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinamide
2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinic acid (30.0 mg,73.6 mol), HATU (33.5 mg,88.3 mol) and N, N-diisopropylethylamine (28.5 mg,220 mol) were dissolved in N, N-dimethylformamide (0.3 mL), and the reaction stirred at room temperature for 30 minutes. Adding 4-amino-bicyclo [2.2.2 to the reaction solution]Octyl-1-alkoxide (15.7 mg,88.3 umol) was stirred at room temperature for 12 hours. The reaction solution was filtered, and the filtrate was concentrated to obtain a crude product. Purification of the crude product by Prep-HPLC gave the title compound (14.5 mg, white solid), yield: 37.11%. MS (ESI) m/z 531.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.40(d,J＝2.4Hz,1H),8.10(s,1H),8.05(d,J＝2.3Hz,1H),7.99(s,1H),7.91(d,J＝8.9Hz,1H),7.80(s,1H),7.71(dd,J＝8.9,1.5Hz,1H),6.89(s,2H),5.44-5.34(m,1H),4.31(s,1H),3.91-3.82(m,2H),3.36-3.29(m,2H),3.16(t,J＝8.8Hz,1H),2.91-2.81(m,2H),2.79-2.77(m,1H),2.43-2.30(m,2H),2.28-2.16(m,1H),2.10-2.02(m,6H),1.81(d,J＝11.6Hz,2H),1.67-1.59(m,6H),1.49-1.36(m,2H).
Example 44
Compound 44: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinamide
(1) 5-bromo-2- (pyrrolidin-3-yl) -2H-indazole
3- (5-bromo-2H-indazol-2-yl) pyrrolidine-1-carboxylic acid tert-butyl ester (1.00 g,2.73mmol, example 43, step 1 isomer) was dissolved in dichloromethane (10 mL) and a hydrochloric acid/dioxane solution (4.0M, 2.05 mL) was slowly added dropwise and reacted at room temperature for 12 hours. The reaction solution was concentrated to give the title compound (700 mg, white solid, hydrochloride). MS (ESI): m/z 266.0[M+H] + . 1 H NMR(400MHz,CD 3 OD)δ8.00(s,1H),7.56(d,J＝1.6Hz,1H),7.23(d,J＝9.0Hz,1H),7.02(dd,J＝9.2,1.8Hz,1H),5.27-5.19(m,1H),3.50-3.45(m,2H),2.97-2.94(m,2H),2.40-2.07(m,2H).
(2) 5-bromo-2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazole
5-bromo-2- (pyrrolidin-3-yl) -2H-indazole (700 mg,2.63mmol, hydrochloride) was dissolved in methanol (7.0 mL) and triethylamine (266 mg,2.63 mmol) and tetrahydro-4H-pyran-4-one (803 mg,3.63 mmol) were added. The reaction solution was stirred at 15℃for 1 hour, and sodium cyanoborohydride (333 mg,5.31 mmol) and acetic acid (175 mg,2.92 mmol) were added. The reaction mixture was stirred at 15℃for a further 12 hours. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL. Times.2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (dichloromethane/methanol=15/1 to 5/1) to give the title compound (330 mg, colorless oil), yield: 35.82%. MS (ESI) m/z 350.1[ M+H ]] + . 1 H NMR(400MHz,CDCl 3 )δ8.03(s,1H),7.74(d,J＝1.2Hz,1H),7.50(d,J＝9.2Hz,1H),7.26(dd,J＝9.1,1.7Hz,1H),5.18-5.05(m,1H),3.97-3.90(m,2H),3.39-3.31(m,2H),3.16-3.03(m,3H),2.75-2.72(m,1H),2.59-2.37(m,2H),2.27-2.18(m,1H),1.80-1.76(m,2H),1.68-1.55(m,2H).
(3) 2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid methyl ester
5-bromo-2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazole (300 mg,856 umol) and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (284 mg,1.03 mmol) were dissolved in 1, 4-dioxane (3.0 mL) and water (0.6 mL), and potassium carbonate (236 mg,1.71 mmol) was added. The system was replaced three times with nitrogen and Pd (dppf) Cl was added under nitrogen blanket 2 (62.6 mg,85.6 umol). The reaction solution was stirred at 100℃for 12 hours. After the reaction mixture was cooled to room temperature, water (5 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 mL. Times.2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product. Purification of the crude product by Prep-TLC gave the title compound (200 mg, yellow oil), yield: 55.40%. MS (MS)(ESI):m/z 422.3[M+H] + . 1 H NMR(400MHz,CDCl 3 )δ8.46(d,J＝2.4Hz,1H),8.32(d,J＝2.4Hz,1H),8.14(s,1H),7.72-7.68(m,2H),7.42-7.38(m,1H),6.34(brs,2H),5.17-5.08(m,1H),3.97-3.90(m,2H),3.86(s,3H),3.42-3.34(m,3H),3.12-3.03(m,2H),2.72-2.50(m,2H),2.42-2.20(m,2H),1.80-1.76(m,2H),1.65-1.53(m,2H).
(4) 2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinate (200 mg,474 umol) was dissolved in methanol (2.0 mL) and water (2.0 mL), lithium hydroxide (22.7 mg,949 umol) was added. The reaction solution was stirred at room temperature for 12 hours. The reaction was concentrated to give crude product, which was purified by Prep-HPLC to give the title compound (100 mg, yellow solid), yield: 51.72%. MS (ESI) m/z 408.2[ M+H ]] + .
(5) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinamide
2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid (50.0 mg, 122. Mu. Mol), HATU (55.9 mg, 147. Mu. Mol) and N, N-diisopropylethylamine (47.5 mg, 365. Mu. Mol) were dissolved in N, N-dimethylformamide (0.5 mL), and the reaction was stirred at room temperature for 30 minutes. Adding 4-amino-bicyclo [2.2.2 to the reaction solution ]Octyl-1-alkoxide (26.1 mg, 147. Mu. Mol) was reacted at room temperature for 12 hours. The reaction solution is filtered, and the filtrate is concentrated to obtain crude products. Purification of the crude product by Prep-HPLC gave the title compound (27.7 mg, pale yellow solid), yield: 42.54%. MS (ESI) m/z 531.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.47(s,1H),8.38(d,J＝2.3Hz,1H),8.03(d,J＝2.3Hz,1H),7.91(s,1H),7.81(s,1H),7.71-7.65(m,1H),7.57(dd,J＝9.0,1.5Hz,1H),6.87(s,2H),5.23-5.20(m,1H),4.30(s,1H),3.90-3.82(m,2H),3.36-3.29(m,2H),3.15-3.04(m,1H),3.02-2.86(m,2H),2.74-2.63(m,1H),2.47-2.40(m,1H),2.38-2.17(m,2H),2.10-2.00(m,6H),1.81(d,J＝11.7Hz,2H),1.67-1.58(m,6H),1.49-1.36(m,2H).
Example 45
Compound 45: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) -1H-indazol-5-yl) nicotinamide
(1) 5-bromo-1- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) -1H-indazole
The procedure of example 41, step (3), was followed using 5-bromo-1- (piperidin-4-yl) -1H-indazole (400 mg,1.26mmol, hydrochloride) and tetrahydro-4H-pyran-4-one (174.40 mg,1.74 mmol) as starting material to give the title compound (220 mg, white solid), yield: 47.8%. 1 H NMR(400MHz,CDCl 3 )δ7.85(s,1H),7.80(d,J＝1.2Hz,1H),7.38-7.33(m,1H),7.31-7.27(m,1H),4.34-4.28(m,1H),3.98(dd,J＝11.1,4.0Hz,2H),3.37-3.30(m,2H),3.08-3.05(m,2H),2.53-2.50(m,1H),2.41-2.18(m,4H),1.97(d,J＝11.5Hz,2H),1.72(d,J＝11.9Hz,2H),1.60-1.56(m,2H).
(2) 2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) -1H-indazol-5-yl) nicotinic acid methyl ester
Methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (100 mg,359.57 mol) and 5-bromo-1- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) -1H-indazole (100 mg,274.51 mol) were dissolved in a mixed solvent of 1, 4-dioxane (2 mL) and water (0.4 mL). Potassium carbonate (75.88 mg,549.03 umol) and Pd (dppf) Cl were added to the reaction system under nitrogen protection 2 (20.09 mg,27.45 umol). The reaction solution was replaced with nitrogen bubbling, and then heated to 110℃and stirred under nitrogen protection for 16 hours. The reaction mixture was cooled to room temperature, concentrated, diluted with methanol (2 mL), and filtered through a syringe filter. The filtrate was purified by Prep-TLC (dichloromethane/methanol=5/1) to give the title compound (80 mg, yellow solid), yield: 66.91%. MS (ESI) m/z 436.3[ M+H ]] + . 1 H NMR(400MHz,CDCl 3 )δ8.45(d,J＝2.6Hz,1H),8.31(d,J＝2.4Hz,1H),7.96(s,1H),7.77(d,J＝1.2Hz,1H),7.50-7.43(m,2H),6.36(brs,2H),4.40-4.34(m,1H),3.99(dd,J＝11.0,3.8Hz,2H),3.86(s,3H),3.38-3.31(m,2H),3.09(d,J＝9.2Hz,2H),2.60-2.47(m,1H),2.44-2.23(m,4H),2.02(d,J＝11.9Hz,2H),1.74(d,J＝11.1Hz,2H),1.66-1.54(m,2H).
(3) 2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) -1H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) -1H-indazol-5-yl) nicotinate (80 mg,183.69 mol) was dissolved in methanol (2.4 mL) and water (2.4 mL), lithium hydroxide monohydrate (16.00 mg,381.28 mol) was added, and the reaction solution was stirred at 20℃for 16 hours. The reaction mixture was concentrated, ethyl acetate (3 mL) and water (5 mL) were added thereto, the aqueous phase was separated, the pH of the aqueous phase was adjusted to 3 to 4 with 1M dilute hydrochloric acid, and a colloidal solid was precipitated and filtered. The filter cake was dried to give the title compound (50 mg, off-white solid), yield: 64.58%. MS (ESI) M/z422.2[ M+H ]] + .
(4) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) -1H-indazol-5-yl) nicotinamide
2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) -1H-indazol-5-yl) nicotinic acid (30 mg, 71.18. Mu. Mol) and 4-aminobicyclo [ 2.2.2.2 ]Octyl-1-alkoxide (15 mg, 84.43. Mu. Mol) was dissolved in N, N-dimethylformamide (0.5 mL), and HATU (32.48 mg, 85.41. Mu. Mol) and N, N-diisopropylethylamine (29.68 mg, 229.64. Mu. Mol) were added. The reaction solution was stirred at 15℃for 2 hours. The reaction was purified by Prep-HPLC to give the title compound (21.6 mg, white solid), yield: 55.06%. MS (ESI) m/z 545.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 ) Delta 8.39 (d, j=2.3 hz, 1H), 8.10 (s, 1H), 8.04 (d, j=2.4 hz, 1H), 7.98 (d, j=0.8 hz, 1H), 7.83-7.76 (m, 2H), 7.69 (dd, j=8.8, 1.7hz, 1H), 6.88 (s, 2H), 4.67-4.56 (m, 1H), 4.30 (s, 1H), 3.92 (dd, j=10.4, 3.3hz, 2H), 3.31-3.27 (m, 2H), 3.04 (d, j=11.4 hz, 2H), 2.57-2.56 (m, 1H), 2.44-2.37 (m, 2H), 2.17-2.01 (m, 8H), 1.94 (d, j=10.0 hz, 2H), 1.72 (d, j=10.4, 3.3.3 hz, 2H), 3.31-3.27 (m, 2H), 3.44-2.37 (d, j=11.4 hz, 2H), 2.17-2.01 (m, 1H), 1.48 (1H)
Compound 46: 2-amino-N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) -5- (2- (1- (oxa-3-yl) piperidin-4-yl) -2H-indazol-5-yl) nicotinamide
(1) 5-bromo-2- (piperidin-4-yl) -2H-indazole
Tert-butyl 4- (5-bromo-2H-indazol-2-yl) piperidine-1-carboxylate (1.2 g,3.16mmol, example 41, step 1 isomer) was dissolved in dichloromethane (5 mL) and a solution of hydrochloric acid/dioxane (4M, 24.00 mL) was added. The reaction solution was stirred at 15℃for 16 hours. The reaction solution was directly concentrated at 45℃to give the title compound (1 g, white solid, hydrochloride) and the crude product was directly used in the next step. MS (ESI) m/z 280.1[ M+H ] ] + .
(2) 5-bromo-2- (1- (oxetan-3-yl) piperidin-4-yl) -2H-indazole
3-oxetanone (150.21 mg,2.09 mmol) was dissolved in methanol (5 mL), triethylamine (159.80 mg,1.58 mmol) and 5-bromo-2- (piperidin-4-yl) -2H-indazole (500.00 mg,1.58mmol, hydrochloride) were added and the reaction stirred at 25℃for 1 hour. Sodium cyanoborohydride (199.99 mg,3.18 mmol) and acetic acid (105.00 mg,1.75 mmol) were further added, and the reaction was stirred at 25℃for a further 15 hours. To the reaction solution was added water (5 mL), extracted with ethyl acetate (5 mL. Times.4), and the organic phases were combined and concentrated. Purification of the crude product by Prep-TLC (dichloromethane/methanol=10/1) gave the title compound (150 mg, white solid), yield: 28.24%. MS (ESI) m/z 336.1[ M+H ]] + . 1 H NMR(400MHz,CDCl 3 )δ7.85(s,1H),7.75(s,1H),7.51(d,J＝9.1Hz,1H),7.26(d,J＝9.1Hz,1H),4.66-4.55(m,4H),4.44-4.33(m,1H),3.53-3.46(m,1H),2.86(d,J＝11.3Hz,2H),2.27-2.09(m,4H),2.06-1.96(m,2H).
(3) 2-amino-5- (2- (1- (oxetan-3-yl) piperidin-4-yl) -2H-indazol-5-yl) nicotinic acid methyl ester
5-bromo-2- (1- (oxetan-3-yl) piperidin-4-yl) -2H-indazole (100 mg, 297.42. Mu. Mol) and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (110 mg, 395.53. Mu. Mol) were dissolved in a mixed solvent of 1, 4-dioxane (2 mL) and water (0.4 mL), and potassium carbonate (82.21 mg, 594.84. Mu. Mol) and Pd (dppf) Cl were added 2 (21.76 mg,29.74 umol). Nitrogen was bubbled into the reaction mixture, and the temperature was raised to 110℃and stirred for 16 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated to give a crude product. Purification of the crude product by Prep-TLC (petroleum ether/methanol=10/1) gave the title compound (60 mg, yellow solid), yield: 49.51%. MS (ESI) m/z 408.2[ M+H ]] + .
(4) 2-amino-5- (2- (1- (oxetan-3-yl) piperidin-4-yl) -2H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (2- (1- (oxetan-3-yl) piperidin-4-yl) -2H-indazol-5-yl) nicotinate (60 mg,147.25 mol) was dissolved in methanol (1 mL) and water (1 mL), and lithium hydroxide monohydrate (12.36 mg,294.50 mol) was added. The reaction solution was stirred at 20℃for 16 hours. The reaction mixture was concentrated, then water (3 mL) and ethyl acetate (1 mL) were added thereto, and the pH of the aqueous phase was adjusted to 3 to 4 with 1M dilute hydrochloric acid. The resulting solution was lyophilized to give a yellow solid. The solid was dispersed in methanol (5 mL) and stirred for 2 hours, then filtered. The filtrate was concentrated to give the title compound (30 mg, yellow solid) and the crude product was used directly in the next step. MS (ESI) m/z 394.2[ M+H ]] + .
(5) 2-amino-N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) -5- (2- (1- (oxa-3-yl) piperidin-4-yl) -2H-indazol-5-yl) nicotinamide
2-amino-5- (2- (1- (oxetan-3-yl) piperidin-4-yl) -2H-indazol-5-yl) nicotinic acid (30 mg,76.25 umol), 4-aminobicyclo [2.2.2]Octyl-1-alkoxide (15 mg, 84.43. Mu. Mol) and HATU (35 mg, 92.05. Mu. Mol) were dissolved in N, N-dimethylformamide (1 mL), and N, N-diisopropylethylamine (29.68 mg, 229.64. Mu. Mol) was added. The reaction solution was stirred at 15℃for 16 hours. Methanol (0.5 mL) was added to the reaction solution for dilution, and the solution was purified by Prep-HPLC to give the title compound (3.7 mg, brown solid), yield: 9.39%. MS (ESI) m/z 517.3[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.46(s,1H),8.38(d,J＝1.5Hz,1H),8.03(s,1H),7.90(s,1H),7.82(s,1H),7.71-7.66(m,1H),7.57(d,J＝8.8Hz,1H),6.87(s,2H),4.61-4.55(m,2H),4.54-4.52(m,1H),4.50-4.43(m,2H),4.30(s,1H),3.51-3.44(m,1H),2.86(d,J＝10.4Hz,2H),2.20-2.10(m,4H),2.09-2.00(m,8H),1.66-1.59(m,6H).
Example 47
Compound 47: 2-amino-5- (1- (4- (oxetan-3-yl) piperazin-1-yl) -2, 3-dihydro-1H-inden-5-yl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
1- (oxetan-3-yl) -4- (5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -2, 3-dihydro-1H-inden-1-yl) piperazine (80 mg, 208.16. Mu. Mol, example 42, step 2) was dissolved in a mixed solvent of 1, 4-dioxane (0.8 mL) and water (0.16 mL), and intermediate 4 (62.48 mg, 208.16. Mu. Mol) and potassium carbonate (86.31 mg, 624.48. Mu. Mol) were added. Pd (dppf) Cl was added to the reaction solution under nitrogen protection 2 (15.23 mg,20.82 umol). The reaction was heated to 80℃and stirred for 12 hours under nitrogen. The reaction solution was cooled to room temperature, water (5 mL) and ethyl acetate (5 mL) were added and the mixture was extracted in portions, and the organic phase was concentrated under reduced pressure to give a crude product. Purification of the crude product by Prep-HPLC gave the title compound (12 mg, white solid), yield: 12.07%. MS (ESI) mz478.3 [ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.42-8.38(m,2H),8.16(d,J＝2.3Hz,1H),7.52(s,1H),7.47(d,J＝7.5Hz,1H),7.32(d,J＝7.7Hz,1H),7.13(s,2H),4.54-4.49(m,2H),4.43-4.37(m,2H),4.34-4.28(m,1H),4.06-3.96(m,1H),3.93-3.86(m,2H),3.43-3.36(m,4H),2.97-2.90(m,1H),2.87-2.81(m,1H),2.46-2.36(m,4H),2.30-2.23(m,3H),2.10-2.02(m,2H),1.82-1.75(m,2H),1.64-1.54(m,2H).
Example 48
Compound 48: (R) -5- (4- ((2-methylpyrrolidin-1-yl) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) -1H-pyrazolo [3,4-b ] pyridin-3-amine
(1) (R) -1- (4-bromobenzyl) -2-methylpyrrolidine
The procedure of step (1) was followed in accordance with example 28, starting from 1-bromo-4- (bromomethyl) benzene (2.94 g,11.74 mmol) and (R) -2-methylpyrrolidine (1 g,11.74 mmol), to give the title compound (1.0 g, pale yellow solid) in yield: 33.5%. MS (ESI) m/z 254.0[ M+H ] ] + .
(2) (R) -2-methyl-1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) pyrrolidine
The procedure of step (2) was followed using (R) -1- (4-bromobenzyl) -2-methylpyrrolidine (1.00 g,3.93 mmol) as a starting material to give the title compound (600 mg, brown solid), yield: 53.6%. MS (ESI) m/z 302.2[ M+H ]] + .
(3) 5-bromo-N- (tetrahydro-2H-pyran-4-yl) -1H-pyrazolo [3,4-b ] pyridin-3-amine
Starting 3-amino-5-bromo-1H-pyrazolo [3,4-b]Pyridine (2.00 g,9.39 mmol) was dissolved in glacial acetic acid (20.0 mL), and tetrahydro-4H-pyran-4-one (1.88 g,18.7 mmol) was added to the reaction solution at 50℃and stirred for 30 min. To the reaction mixture was added sodium cyanoborohydride (1.18 g,18.7 mmol) in portions, and the reaction mixture was stirred for an additional 2 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the residue was dissolved in 5% aqueous sodium hydroxide (20 mL) and extracted with dichloromethane (20 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (1.0 g, brown solid), yield: 35.85%. MS (ESI) m/z 297.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ12.23(s,1H),8.43-8.40(m,2H),6.21(d,J＝7.0Hz,1H),3.93-3.87(m,2H),3.74-3.66(m,1H),3.46-3.39(m,2H),2.01(d,J＝11.7Hz,2H),1.52-1.44(m,2H).
(4) (R) -5- (4- ((2-methylpyrrolidin-1-yl) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) -1H-pyrazolo [3,4-b ] pyridin-3-amine
5-bromo-N- (tetrahydro-2H-pyran-4-yl) -1H-pyrazolo [3,4-b]Pyridin-3-amine (100 mg, 336. Mu. Mol), (R) -2-methyl-1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) pyrrolidine (101 mg, 336. Mu. Mol) and sodium carbonate (16.5 mg, 156. Mu. Mol) were dissolved in a mixed solvent of 1, 4-dioxane (1.0 mL) and water (0.1 mL). Pd (dppf) Cl is added into the reaction system under the protection of nitrogen 2 (21.96 mg, 30.01. Mu. Mol) and the reaction mixture was stirred under nitrogen at 90℃for 16 hours. The reaction was cooled to room temperature, diluted with methanol (1.0 mL) and filtered, and the filtrate was purified by Prep-HPLC to give the title compound (10.6 mg, yellow solid), yield: 8.05%. MS (ESI) m/z 392.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ12.04(s,1H),8.68(d,J＝2.1Hz,1H),8.47(d,J＝2.0Hz,1H),7.63(d,J＝8.1Hz,2H),7.41(d,J＝8.0Hz,2H),6.21(d,J＝7.0Hz,1H),4.01(d,J＝13.1Hz,1H),3.95-3.87(m,2H),3.81-3.69(m,1H),3.50-3.38(m,2H),3.17(d,J＝13.1Hz,1H),2.87-2.76(m,1H),2.42-2.40(m,1H),2.15-2.01(m,3H),1.99-1.86(m,1H),1.70-1.57(m,2H),1.56-1.44(m,2H),1.43-1.29(m,1H),1.13(d,J＝6.0Hz,3H).
Example 49
Compound 49: (R) -5- (4- ((2-methylpyrrolidin-1-yl) methyl) phenyl) -3- (1- (tetrahydro-2H-pyran-4-yl) -1H-1,2, 3-triazol-4-yl) pyridin-2-amine
(1) 4-azidothydro-2H-pyranes
4-iodotetrahydro-2H-pyran (1.00 g,3.93 mmol), sodium azide (430 mg,3.93 mmol) is dissolved in N, N-dimethylformamide (10.0 mL) and N, N-diisopropylethylamine (609.54 mg,4.72 mmol) is added. The reaction mixture was heated to 80 ℃ and stirred for 2 hours. The reaction solution was cooled to room temperature, the pH of the reaction solution was adjusted to more than 9 with saturated aqueous sodium bicarbonate, and extraction was performed with ethyl acetate (10 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (600 mg, yellow solid), the crude was used directly in the next step.
(2) 5-bromo-3- (1- (tetrahydro-2H-pyran-4-yl) -1H-1,2, 3-triazol-4-yl) pyridin-2-amine
5-bromo-3-ethynylpyridin-2-amine (100 mg,507 umol) and 4-azidothydro-2H-pyran (129 mg,1.02 mmol) were dissolved in tert-butanol (0.5 mL) and water (1.0 mL) and cuprous iodide (9.67 mg,50.75 umol) was added. The reaction mixture was heated to 80 ℃ and stirred for 16 hours. The reaction mixture was cooled to room temperature, quenched with water (10 mL) and extracted with ethyl acetate (10 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (120 mg, yellow solid), yield: 72.9%. MS (ESI) m/z 324.1[ M+H ]] + .
(3) (R) -5- (4- ((2-methylpyrrolidin-1-yl) methyl) phenyl) -3- (1- (tetrahydro-2H-pyran-4-yl) -1H-1,2, 3-triazol-4-yl) pyridin-2-amine
5-bromo-3- (1- (tetrahydro-2H-pyran)-4-yl) -1H-1,2, 3-triazol-4-yl) pyridin-2-amine (30.0 mg,92.5 mol), (R) -2-methyl-1- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) pyrrolidine (55.7 mg,185 mol, example 48, step 2) and cesium carbonate (90.4 mg,277 mol) were dissolved in 1, 4-dioxane (1.0 mL) and water (0.05 mL). Pd (dppf) Cl was added to the reaction solution under nitrogen protection 2 (6.77 mg,9.25 umol). The reaction mixture was heated to 100 ℃ and stirred for 16 hours. The reaction was cooled to room temperature, diluted with methanol (1.0 mL) and filtered, and the filtrate was purified by Prep-HPLC to give the title compound (8.70 mg, yellow solid), yield: 22.46%. MS (ESI) m/z 419.2[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.00(s,1H),8.32(d,J＝2.4Hz,1H),8.21(d,J＝2.4Hz,1H),7.62(d,J＝8.2Hz,2H),7.36(d,J＝8.1Hz,2H),7.07(s,2H),5.00-4.73(m,1H),4.07-3.95(m,3H),3.56(dt,J＝11.7,2.0Hz,2H),3.16(d,J＝13.1Hz,1H),2.84-2.75(m,1H),2.41-2.39(m,1H),2.20-2.13(m,2H),2.11-2.02(m,3H),1.97-1.85(m,1H),1.66-1.54(m,2H),1.42-1.30(m,1H),1.12(d,J＝6.0Hz,3H).
Example 50
Compound 50:4- (2-amino-5- (4-morpholinophenyl) pyridin-3-yl) -N- (cyanomethyl) benzamide
(1) 4- (2-amino-5-bromopyridin-3-yl) benzoic acid methyl ester
5-bromo-3-iodopyridin-2-amine (500 mg,1.67 mmol) was dissolved in a mixed solvent of toluene (10 mL), ethanol (5 mL) and water (2.5 mL), and 4-methoxycarbonylphenylboronic acid (340 mg,1.89 mmol) and sodium carbonate (637 mg,6.02 mmol) were added sequentially. Pd (PPh) was added to the reaction mixture under nitrogen protection 3 ) 4 (96 mg,83.64 umol). The reaction solution was reacted at 135℃for 0.5 hour under microwave conditions. The reaction was cooled to room temperature, water (15 mL) and ethyl acetate (5 mL) were added to conduct liquid-split extraction, and the organic phase was concentrated to give the title compound (550 mg, yellow solid), and the crude product was used in the next step without purification. MS (ESI) m/z 307.1[ M+H ]] + .
(2) 4- (2-amino-5- (4-morpholinophenyl) pyridin-3-yl) benzoic acid methyl ester
Methyl 4- (2-amino-5-bromopyridin-3-yl) benzoate (500 mg,1.63 mmol) was dissolved in a mixed solvent of ethylene glycol dimethyl ether (5 mL) and water (1 mL), and (4-morpholinophenyl) boric acid (404 mg,1.95 mmol) and sodium carbonate (517 mg,4.88 mmol) were added. Pd (PPh) was added to the reaction mixture under nitrogen protection 3 ) 4 (188 mg,162.79 umol). The reaction was heated to 80℃and stirred for 3 hours under nitrogen. The reaction was cooled to room temperature, water (20 mL) and ethyl acetate (20 mL) were added and the organic phase was concentrated to give the title compound (0.6 g, yellow solid) which was used in the next step without purification. MS (ESI) m/z 390.3[ M+H ] ] + .
(3) 4- (2-amino-5- (4-morpholinophenyl) pyridin-3-yl) benzoic acid
Methyl 4- (2-amino-5- (4-morpholinophenyl) pyridin-3-yl) benzoate (0.3 g,770.32 umol) was dissolved in a mixed solvent of methanol (6 mL) and water (3 mL), sodium hydroxide (61.62 mg,1.54 mmol) was added, and the reaction solution was stirred at 20℃for 12 hours. The reaction solution was concentrated under reduced pressure, water (10 mL) and ethyl acetate (10 mL) were added to the reaction solution, and the organic phase was discarded. The aqueous phase was adjusted to pH 2 with 2M aqueous hydrochloric acid and the aqueous phase was concentrated to a yellow solid. The solid was dispersed in methanol (10 mL), stirred for 2 hours, filtered, and the filtrate was concentrated to give the title compound (150 mg, yellow solid), the crude was used in the next step without purification. MS (ESI) m/z 376.3[ M+H ]] + .
(4) 4- (2-amino-5- (4-morpholinophenyl) pyridin-3-yl) -N- (cyanomethyl) benzamide
4- (2-amino-5- (4-morpholinophenyl) pyridin-3-yl) benzoic acid (100 mg,266.37 mmol) was dissolved in N, N-dimethylformamide (1 mL) and HATU (111 mg,293.01 mmol) and N, N-diisopropylethylamine (172 mg,1.33 mmol) were added sequentially. The reaction solution was stirred at 20℃for 30 minutes, 2-aminoacetonitrile hydrochloride (36 mg,399.55 umol) was added, and stirring was continued for 3 hours. The reaction was filtered and the filtrate purified by Pre-HPLC to give the title compound (3 mg, white solid), yield: 3.0%. MS (ESI) m/z 414.0[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.31-9.23(m,1H),8.27(d,J＝2.3Hz,1H),7.98(d,J＝8.2Hz,2H),7.68(d,J＝8.2Hz,2H),7.60(d,J＝2.3Hz,1H),7.53(d,J＝8.7Hz,2H),7.00(d,J＝8.7Hz,2H),5.74(s,2H),4.35(d,J＝5.4Hz,2H),3.78-3.73(m,4H),3.16-3.11(m,4H).
Example 51
Compound 51:4- (2-amino-5- (4- (morpholinomethyl) phenyl) pyridin-3-yl) -N- (cyanomethyl) benzamide
(1) 4- (2-amino-5- (4- (morpholinomethyl) phenyl) pyridin-3-yl) benzoic acid methyl ester
Referring to the procedure of example 50, step (2), starting from methyl 4- (2-amino-5-bromopyridin-3-yl) benzoate (500 mg,1.63mmol, example 50, step 1) and 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzyl) morpholine (592.31 mg,1.95 mmol), the title compound (600 mg, orange solid) was obtained in yield: 91.4%. MS (ESI) m/z 404.3[ M+H ]] + .
(2) 4- (2-amino-5- (4- (morpholinomethyl) phenyl) pyridin-3-yl) benzoic acid
Referring to the procedure of example 50, step (3), starting from methyl 4- (2-amino-5- (4- (morpholinomethyl) phenyl) pyridin-3-yl) benzoate (350 mg,867.47 mol), the title compound (300 mg, orange solid) was obtained, crude was used directly in the next step without purification. MS (ESI): M/z390.3[ M+H ]] + .
(3) 4- (2-amino-5- (4- (morpholinomethyl) phenyl) pyridin-3-yl) -N- (cyanomethyl) benzamide
Referring to the procedure of example 50, step (4), starting from 4- (2-amino-5- (4- (morpholinomethyl) phenyl) pyridin-3-yl) benzoic acid (150 mg,385.16 mol), the title compound (5 mg, white solid) was obtained in the yield: 3.4%. MS (ESI) m/z 428.3[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.27(t,J＝5.4Hz,1H),8.33(d,J＝2.4Hz,1H),7.99(d,J＝8.3Hz,2H),7.68(d,J＝8.3Hz,2H),7.66(d,J＝2.3Hz,1H),7.61(d,J＝8.2Hz,2H),7.35(d,J＝8.1Hz,2H),5.86(s,2H),4.35(d,J＝5.4Hz,2H),3.58(t,J＝4.4Hz,4H),3.48(s,2H),2.38-2.36(m,4H).
Example 52
Compound 52: n- (cyanomethyl) -4- (6- (4-morpholinophenyl) pyrazolo [1,5-a ] pyrimidin-3-yl) benzamide
(1) 4- (6-Bromopyrazolo [1,5-a ] pyrimidin-3-yl) benzoic acid methyl ester
Reference example 50 procedure of step (1) as 6-bromo-3-iodopyrazolo [1,5-a]Pyrimidine (300 mg,926.17 mmole) and 4-methoxycarbonylphenylboronic acid (188.35 mg,1.05 mmol) were used as starting materials to give the title compound (100 mg, yellow solid), yield: 32.51%. MS (ESI): M/z332.0[ M+H ]] + . 1 H NMR(400MHz,CDCl 3 )δ8.79(d,J＝2.2Hz,1H),8.52(d,J＝2.2Hz,1H),8.40(s,1H),8.05(s,4H),3.87(s,3H).
(2) 4- (6- (4-morpholinophenyl) pyrazolo [1,5-a ] pyrimidin-3-yl) benzoic acid
4- (6-bromopyrazolo [1, 5-a)]Pyrimidine-3-yl) benzoic acid methyl ester (50 mg, 150.53. Mu. Mol) and (4-morpholinophenyl) boronic acid (40.00 mg, 193.21. Mu. Mol) were dissolved in a mixed solvent of 1, 4-dioxane (1 mL) and water (0.5 mL), cesium carbonate (147.14 mg, 451.60. Mu. Mol) and Pd (dppf) Cl were added 2 (11.01 mg,15.05 umol). The reaction solution was replaced with nitrogen three times, and then heated to 110℃and stirred for 16 hours. The reaction solution was cooled to room temperature and concentrated. The crude product was dispersed in methanol (5 mL) and filtered, and the filter cake was dissolved in dimethyl sulfoxide (5 mL) and purified by Prep-HPLC to give the title compound (3 mg, yellow solid), yield: 4.98%. MS (ESI) m/z 401.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ12.80(brs,1H),9.46(d,J＝2.0Hz,1H),9.10(d,J＝2.0Hz,1H),8.88(s,1H),8.33(d,J＝8.3Hz,2H),8.02(d,J＝8.3Hz,2H),7.78(d,J＝8.7Hz,2H),7.11(d,J＝8.7Hz,2H),3.81-3.74(m,4H),3.23-3.19(m,4H).
(3) N- (cyanomethyl) -4- (6- (4-morpholinophenyl) pyrazolo [1,5-a ] pyrimidin-3-yl) benzamide
Reference example 50 procedure of step (4) was followed as 4- (6- (4-morpholinophenyl) pyrazolo [1, 5-a)]Pyrimidine-3-yl) benzoic acid (3 mg,7.49 umol) as a starting material,the title compound (1 mg, yellow solid) was obtained in the yield: 30.44%. MS (ESI) m/z 439.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ9.46(d,J＝2.4Hz,1H),9.20(brs,1H),9.11(d,J＝2.3Hz,1H),8.88(s,1H),8.33(d,J＝8.6Hz,2H),7.97(d,J＝8.6Hz,2H),7.78(d,J＝8.8Hz,2H),7.11(d,J＝8.8Hz,2H),4.34(d,J＝4.8Hz,2H),3.81-3.75(m,4H),3.23-3.20(m,4H).
Example 53
Compound 53: (R) -2-amino-5- (2-fluoro-4- (2-methylpyrrolidin-1-yl) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
In a similar manner to that of example 22, starting from 4-bromo-3-fluorobenzaldehyde and (R) -2-methylpyrrolidine in step 1, the title compound (13.0 mg, white solid) was obtained. MS (ESI) m/z 413.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.35(d,J＝7.6Hz,1H),8.26(d,J＝2.0Hz,1H),8.07(d,J＝2.0Hz,1H),7.50(t,J＝8.0Hz,1H),7.23-7.19(m,4H),4.02-3.94(m,2H),3.87(dd,J＝12.0,2.4Hz,2H),3.41-3.35(m,2H),3.20(d,J＝13.6Hz,1H),2.83-2.78(m,1H),2.45-2.93(m,1H),2.09(q,J＝8.8Hz,1H),1.94-1.88(m,1H),1.78-1.74(m,2H),1.67-1.50(m,4H),1.40-1.31(m,1H),1.10(d,J＝6.0Hz,3H).
Example 54
Compound 54:5- (4- (2-oxo-6-azaspiro [3.3] heptan-6-yl) phenyl) -2-amino-N- (tetrahydro-2H-pyran-4-yl) nicotinamide
Referring to the procedure of example 30, step 4 was followed by 5- (4- (2-oxa-6-azaspiro [3.3 ]) step]Heptane-6-yl) phenyl) -2-aminonicotinic acid (35 mg,0.11 mmol) and tetrahydro-2H-pyran-4-amine (34 mg,0.34 mmol) as starting material gave the title compound (15 mg, light yellow solid), yield: 34.6%. MS (ESI): m/z 395.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.38(d,J＝8.0Hz,1H),8.30(d,J＝2.0Hz,1H),8.06(d,J＝2.4Hz,1H),7.48(d,J＝8.8Hz,2H),6.99(s,2H),7.51(d,J＝8.4Hz,2H),4.73(s,4H),4.02-3.96(m,5H),3.91-3.87(m,2H),3.41-3.35(m,2H),1.79-1.75(m,2H),1.62-1.52(m,2H).
Example 55
Compound 58: (R) -2-amino-5- (2-fluoro-4- (2-methylpyrrolidin-1-yl) methyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
Referring to the procedure of example 22, step 1 was conducted using 4-bromo-3-fluorobenzaldehyde and (R) -2-methylpyrrolidine as starting materials, and step 3 was conducted using (R) -2-amino-5- (2-fluoro-4- ((2-methylpyrrolidin-1-yl) methyl) phenyl) nicotinic acid and ((2 s, 5R) -5-aminotetrahydro-2H-pyran-2-yl) methanol hydrochloride as starting materials, to give the title compound (10.0 mg, white solid) through similar procedures. MS (ESI) m/z 443.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.27-8.21(m,2H),8.06(d,J＝1.2Hz,1H),7.49(t,J＝8.0Hz,1H),7.23-7.18(m,4H),4.62(t,J＝6.0Hz,1H),3.98(d,J＝13.6Hz,1H),3.89-3.86(m,2H),3.40-3.36(m,1H),3.24-3.08(m,4H),2.82-2.78(m,1H),2.43-2.38(m,1H),2.08(q,J＝8.8Hz,1H),1.97-1.90(m,2H),1.74-1.54(m,4H),1.38-1.29(m,2H),1.10(d,J＝5.6Hz,3H).
Example 56
Compound 88: 2-amino-5- (2- (1- (4, 4-difluorocyclohexyl) pyrrolidin-3-yl) -2H-indazol-5-yl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) nicotinamide
(1) 3- (5-bromo-2H-indazol-2-yl) pyrrolidine-1-carboxylic acid tert-butyl ester
5-bromo-1H-indazole (3.20 g,16.3 mmol), 3- ((methylsulfonyl) oxy) pyrrolidine-1-carboxylic acid tert-butyl ester (5.19 g,19.6 mmol), potassium carbonate (4.50 g,32.6 mmol) was dissolved in N, N-dimethylformamide (30 mL). The temperature of the reaction solution is raised to 100Stirred overnight at c. Water (50 mL) was added to the reaction system, the mixture was extracted with ethyl acetate (20 mL. Times.2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give tert-butyl 3- (5-bromo-1H-indazol-1-yl) pyrrolidine-1-carboxylate (1.2 g, yellow solid, yield: 20.1%) and tert-butyl 3- (5-bromo-2H-indazol-2-yl) pyrrolidine-1-carboxylate (1.0 g, yellow solid, yield: 16.8%). MS (ESI) m/z 310.0[ M+H-56 ] ] + .
(2) 5-bromo-2- (pyrrolidin-3-yl) -2H-indazole hydrochloride
3- (5-bromo-2H-indazol-2-yl) pyrrolidine-1-carboxylic acid tert-butyl ester (600 mg,1.64 mmol) was dissolved in dichloromethane (5 mL), 4N dioxane hydrochloride solution (4 mL) was added, stirring was continued for 3 hours, and a large amount of solid was precipitated. The reaction solution was dried by spinning to give the title compound (436 mg, white solid). MS (ESI) m/z 266.0[ M+H ]] + .
(3) 5-bromo-2- (1- (4, 4-difluorocyclohexyl) pyrrolidin-3-yl) -2H-indazole
5-bromo-2- (pyrrolidin-3-yl) -2H-indazole hydrochloride (436 mg,1.6 mmol) was dissolved in dichloromethane (10 mL), 4-difluorocyclohexanone (322 mg,2.4 mmol) and 2 drops of acetic acid were added and the reaction stirred for 20 minutes. Sodium cyanoborohydride (202 mg,3.2 mmol) was added and stirring continued overnight. To the reaction was added water (20 mL), the mixture was extracted with dichloromethane (20 ml×2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=30/1) to give the title compound (321 mg, yellow oil), yield: 51%. MS (ESI) m/z 384.0[ M+H ]] + .
(4) 2-amino-5- (2- (1- (4, 4-difluorocyclohexyl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid methyl ester
5-bromo-2- (1- (4, 4-difluorocyclohexyl) pyrrolidin-3-yl) -2H-indazole (250 mg,0.6 mmol) was dissolved in 1, 4-dioxane (3 mL) and water (0.3 mL), and (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boronic acid (158 mg,0.8 mmol), potassium carbonate (221 mg,1.6 mmol) and Pd (dppf) Cl were added 2 (44 mg,0.06 mmol). The mixture was stirred at reflux under nitrogen overnight. Water (20 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (20 mL. Times.2) without any reactionDrying over sodium sulfate, filtration, and concentration under reduced pressure, the crude product was purified by column chromatography (dichloromethane/methanol=40/1) to give the title compound (200 mg, yellow oil), yield: 67%. MS (ESI) m/z 456.2[ M+H ]] + .
(5) 2-amino-5- (2- (1- (4, 4-difluorocyclohexyl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (2- (1- (4, 4-difluorocyclohexyl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinate (200 mg,0.44 mmol) was dissolved in a mixed solvent of methanol (2 mL) and water (2 mL), and sodium hydroxide (53 mg,1.32 mmol) was added. The reaction mixture was heated to 50℃and reacted for 3 hours. Concentrated under reduced pressure, water was added, the aqueous phase was adjusted to pH 2 with 1N hydrochloric acid, and lyophilized to give the title compound (194 mg, gray solid). MS (ESI): M/z440.2[ M-H ]] - .
(6) 2-amino-5- (2- (1- (4, 4-difluorocyclohexyl) pyrrolidin-3-yl) -2H-indazol-5-yl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) nicotinamide
2-amino-5- (2- (1- (4, 4-difluorocyclohexyl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid (50 mg,0.11 mmol) was dissolved in N, N-dimethylformamide (3 mL) and HATU (84 mg,0.22 mmol), N, N-diisopropylethylamine (43 mg,0.33 mmol) and 4-aminobicyclo [ 2.2.2.2 were added ]Octane-1-hydrochloride (22 mg,0.12 mmol). The mixture was stirred at room temperature overnight. The reaction was purified by Prep-HPLC to give the title compound (0.8 mg, white solid), yield: 1.2%. MS (ESI) m/z 565.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.45(s,1H),8.38(d,J＝2.0Hz,1H),8.03(d,J＝2.4Hz,1H),7.91(s,1H),7.81(s,1H),7.67(d,J＝9.2Hz,1H),7.57(d,J＝8.8Hz,1H),6.86(s,2H),5.25-5.20(m,1H),4.30(s,1H),3.12-3.08(m,1H),3.02-2.96(m,2H),2.74-2.66(m,1H),2.35-2.33(m,1H),2.27-2.21(m,1H),2.08-2.00(m,9H),1.85-1.82(m,4H),1.65-1.61(m,8H).
Example 57
Compound 115: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinamide
Reference example 67 procedure 8 was followed using 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine)]-5-yl) nicotinic acid and 4-amino-bicyclo [2.2.2]Octane-1-hydrochloride as a starting material gave the title compound (4.7 mg, white solid). MS (ESI): M/z517.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.32(d,J＝2.4Hz,1H),7.97(d,J＝2.4Hz,1H),7.76(s,1H),7.45-7.43(m,2H),7.33(d,J＝8.4Hz,1H),6.89(s,2H),4.32(s,1H),3.88-3.84(m,2H),3.33-3.30(m,2H),2.89(t,J＝7.2Hz,2H),2.78(t,J＝6.8Hz,2H),2.73-2.60(m,2H),2.34-2.24(m,1H),2.17-2.10(m,1H),2.09-2.01(m,7H),1.97-1.92(m,2H),1.80-1.77(m,2H),1.65-1.61(m,6H),1.47-1.39(m,2H).
Example 58
Compound 124: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (4-morpholinopiperidin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinamide
(1) 4- (1- (5-bromo-2, 3-dihydro-1H-inden-1-yl) piperidin-4-yl) morpholine
4- (piperidin-4-yl) morpholine (408 mg,2.4 mmol), 5-bromo-2, 3-dihydro-1H-inden-1-one (500 mg,2.4 mmol) was dissolved in tetrahydrofuran (10 mL), and tetraethyltitanate (602 mg,2.6 mmol) was added. The mixture was stirred at 60℃for 1 hour, then sodium cyanoborohydride (302 mg,4.8 mmol) and ethanol (10 mL) were added and stirring was continued at room temperature overnight. Water (20 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (40 mL. Times.2) in portions, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane/methanol=20/1) to give the title compound (170 mg, yellow oil), yield: 20%. MS (ESI) m/z 365.0[ M+H ] ] + .
(2) 2-amino-5- (1- (4-morpholinopiperidin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinic acid methyl ester
4- (1- (5-bromo-2, 3-dihydro-1H-inden-1-yl) piperidin-4-yl) morpholine (100 mg,0.3 mmol), (6-amino-5- (methoxycarbonyl) pyridine-3-yl) boronic acid (83 mg,0.3 mmol), pdCl 2 (dppf) (24 mg,0.03 mmol), potassium carbonate (83 mg,0.6 mmol) was dissolved in a mixed solvent of 1, 4-dioxane/water (5 mL/1 mL). The mixture was stirred overnight at 100℃under nitrogen. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (100% ethyl acetate) to give the title compound (60 mg, colorless oil), yield: 50%. MS (ESI): m/z 437.2[ M+H ]] + .
(3) 2-amino-5- (1- (4-morpholinopiperidin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinic acid
Methyl 2-amino-5- (1- (4-morpholinopiperidin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinate (60 mg,0.1 mmol) was dissolved in a mixed solvent of methanol/water (5 mL/5 mL), sodium hydroxide (12 mg,0.3 mmol) was added, and the reaction was stirred at 50℃overnight. The reaction was dried with screw to adjust the pH to 3-4 with dilute hydrochloric acid, concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (40 mg, colorless oil), yield: 69%. MS (ESI) m/z 423.2[ M+H ]] + .
(4) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (4-morpholinopiperidin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinamide
2-amino-5- (1- (4-morpholinopiperidin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinic acid (40 mg,0.09 mmol), 4-amino-bicyclo [ 2.2.2.2]Octane-1-hydrochloride (17 mg,0.09 mmol) was dissolved in N, N-dimethylformamide (5 mL), and HATU (51 mg,0.14 mmol) and N, N-diisopropylethylamine (23 mg,0.18 mmol) were added to the reaction solution and stirred at room temperature overnight. The reaction was directly purified by Pre-HPLC to give the title compound (10 mg, white solid) in 19% yield. MS (ESI) m/z 546.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.33(d,J＝2.4Hz,1H),7.99(d,J＝2.4Hz,1H),7.76(s,1H),7.47-7.43(m,2H),7.30(d,J＝8.0Hz,1H),6.90(s,2H),4.32-4.28(m,2H),3.56-3.54(m,4H),2.95-2.78(m,3H),2.45-2.44(m,4H),2.27-2.22(m,1H),2.12-2.02(m,9H),1.79-1.69(m,2H),1.64-1.60(m,6H),1.47-1.24(m,4H).
Example 59
Compound 125: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (4- (tetrahydro-2H-pyran-4-yl) piperazin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinamide
(1) 4- (5-bromo-2, 3-dihydro-1H-inden-1-yl) piperazine-1-carboxylic acid tert-butyl ester
5-bromo-2, 3-dihydro-1H-inden-1-one (3.00 g,14.36 mmol) and piperazine-1-carboxylic acid tert-butyl ester (2.94 g,15.79 mmol) were dissolved in tetrahydrofuran (30 mL), and tetraethyltitanate (3.6 g,15.79 mmol) was added. The reaction mixture was warmed to 60℃and stirred for 2 hours, cooled to room temperature, and ethanol (75 mL) and sodium cyanoborohydride (1.81 g,28.72 mmol) were added thereto, and the reaction was continued at room temperature overnight under nitrogen protection. The reaction mixture was quenched with water (50 mL), filtered through celite, the filtrate concentrated under reduced pressure, extracted with ethyl acetate (50 mL. Times.3), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by column chromatography (dichloromethane/methanol=100/1) to give the title compound (970 mg, brown oil), yield: 17.8%. MS (ESI) m/z 381.1[ M+H ] ] + .
(2) 1- (5-bromo-2, 3-dihydro-1H-inden-1-yl) piperazine
4- (5-bromo-2, 3-dihydro-1H-inden-1-yl) piperazine-1-carboxylic acid tert-butyl ester (970 mg,2.56 mmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (3 mL) was added and reacted at room temperature for 2 hours. The reaction was neutralized with saturated sodium bicarbonate solution, extracted with dichloromethane (15 ml x 3), dried over anhydrous sodium sulfate, and concentrated to give the title compound (700 mg, brown oil) which was used directly in the next step. MS (ESI) m/z 281.1[ M+H ]] + .
(3) 1- (5-bromo-2, 3-dihydro-1H-inden-1-yl) -4- (tetrahydro-2H-pyran-4-yl) piperazine
1- (5-bromo-2, 3-dihydro-1H-inden-1-yl) piperazine (500 mg,1.79 mmol) and tetrahydro-4H-pyran-4-one (200 mg,1.97 mmol) were dissolved in tetrahydrofuran (20 mL), and tetraethyltitanate (450 mg,1.97 mmol) was added. The reaction mixture was warmed to 60℃and stirred for 2 hours, cooled to room temperature, ethanol (30 mL) and sodium cyanoborohydride (226 mg,3.58 mmol) were added, and the reaction was continued at room temperature overnight under nitrogen protection. The reaction was quenched with water (20 mL), filtered through celite, the filtrate concentrated under reduced pressure, extracted with ethyl acetate (20 mL. Times.3), dried over anhydrous sodium sulfate, and concentrated. Residue warpPurification by column chromatography (dichloromethane/methanol=10/1) gave the title compound (90 mg, white solid) in a yield of 11.8%. MS (ESI) m/z 365.1[ M+H ] ] + .
(4) 2-amino-5- (1- (4- (tetrahydro-2H-pyran-4-yl) piperazin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinic acid methyl ester
1- (5-bromo-2, 3-dihydro-1H-inden-1-yl) -4- (tetrahydro-2H-pyran-4-yl) piperazine (90 mg,0.25 mmol) and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (83.4 mg,0.30 mmol) were dissolved in a mixed solvent of 1, 4-dioxane/water (2 mL/0.5 mL) and [1,1' -bis (diphenylphosphine) ferrocene was added]Palladium dichloride (22 mg,0.03 mmol) and potassium carbonate (103.5 mg,0.75 mmol). The reaction solution was replaced with nitrogen, heated to 80℃and stirred for 16 hours. The reaction was concentrated, and the residue was purified by column chromatography (dichloromethane/methanol=100/1 to 10/1) to give the title compound (70 mg, orange oil), yield: 64.2%. MS (ESI): m/z 437.2[ M+H ]] + .
(5) 2-amino-5- (1- (4- (tetrahydro-2H-pyran-4-yl) piperazin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinic acid
Methyl 2-amino-5- (1- (4- (tetrahydro-2H-pyran-4-yl) piperazin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinate (70 mg,0.16 mmol) was dissolved in a mixed solution of methanol/water (2 mL/2 mL), sodium hydroxide (20 mg,0.48 mmol) was added, and the reaction was heated to 50℃and stirred for 2 hours. Methanol was removed by concentration under reduced pressure, pH was adjusted to neutrality with 2N hydrochloric acid solution, and purified by reverse phase column to give the title compound (30 mg, white solid), yield: 44.4%. MS (ESI) m/z 423.2[ M+H ] ] + .
(6) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (4- (tetrahydro-2H-pyran-4-yl) piperazin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinamide
2-amino-5- (1- (4- (tetrahydro-2H-pyran-4-yl) piperazin-1-yl) -2, 3-dihydro-1H-inden-5-yl) nicotinic acid (30 mg,0.07 mmol) was dissolved in N, N-dimethylformamide (1 mL), HATU (53.2 mg,0.14 mmol) and N, N-diisopropylethylamine (27.3 mg,0.21 mmol) were added followed by 4-aminobicyclo [ 2.2.2.2]Octane-1-hydrochloride (13.84 mg,0.08 mmol), the reaction was stirred at room temperature for 3 hours. The reaction solution was purified by Pre-HPLC to give the title compound (10.9 mg, white solid)) Yield: 28.6%. MS (ESI) m/z 546.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.33(d,J＝2.4Hz,1H),7.99(d,J＝2.4Hz,1H),7.76(s,1H),7.48(s,1H),7.44(d,J＝8.0Hz,1H),7.30(d,J＝7.6Hz,1H),6.90(s,2H),4.31(s,1H),4.26(t,J＝7.2Hz,1H),3.86(d,J＝8.8Hz,2H),3.25(t,J＝11.6Hz,2H),2.96-2.88(m,1H),2.84-2.77(m,1H),2.48-2.29(m,7H),2.06-2.00(m,8H),1.69-1.60(m,8H),1.40-1.24(m,4H).
Example 60
Compound 126: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (tetrahydro-2H-pyran-4-yl) -1H-indazol-5-yl) nicotinamide
(1) 5-bromo-1- (tetrahydro-2H-pyran-4-yl) -1H-indazole
5-bromo-1H-indazole (1.5 g,7.6 mmol), tetrahydro-2H-pyran-4-ylmethane sulfonate (2.0 g,11.4 mmol), cesium carbonate (4.96 g,15.2 mmol) was dissolved in N, N-dimethylformamide (20 mL). The temperature was raised to 100℃and the reaction was stirred overnight. To the reaction was added water (50 mL), and the mixture was extracted with ethyl acetate (20 ml×2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give 5-bromo-1- (tetrahydro-2H-pyran-4-yl) -1H-indazole (1.6 g, yellow solid, yield: 49%) and 5-bromo-2- (tetrahydro-2H-pyran-4-yl) -2H-indazole (1.2 g, yellow solid, yield: 38%). MS (ESI) m/z 281.0[ M+H ] ] + .
(2) 1- (tetrahydro-2H-pyran-4-yl) -5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indazole
5-bromo-1- (tetrahydro-2H-pyran-4-yl) -1H-indazole (100 mg,0.357 mmol) was dissolved in 1, 4-dioxane (2 mL), pinacol biborate (136 mg,0.54 mmol), potassium acetate (88 mg,1.07 mmol) and Pd (dppf) Cl were added 2 (30 mg,0.041 mmol). The mixture was stirred at reflux under nitrogen overnight. Water (50. 50 mL) was added to the reaction system, the mixture was extracted with ethyl acetate (20 mL. Times.2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered,concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give the title compound (30 mg), yield: 25.6%. MS (ESI) m/z 329.2[ M+H ]] + .
(3) 2-amino-5- (1- (tetrahydro-2H-pyran-4-yl) -1H-indazol-5-yl) nicotinic acid methyl ester
1- (tetrahydro-2H-pyran-4-yl) -5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indazole (118 mg, 0.317 mmol) was dissolved in a mixed solvent of 1, 4-dioxane (2 mL) and water (0.2 mL), methyl 2-amino-5-bromonicotinate (99 mg,0.429 mmol), potassium carbonate (148 mg,1.07 mmol) and Pd (dppf) Cl were added 2 (30 mg,0.041 mmol). The mixture was stirred at reflux under nitrogen overnight. To the reaction system was added water (50 mL), the mixture was extracted with ethyl acetate (20 ml×2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography (dichloromethane/methanol=30/1) to give the title compound (100 mg), yield: 79%. MS (ESI) m/z 353.2[ M+H ] ] + .
(4) 2-amino-5- (1- (tetrahydro-2H-pyran-4-yl) -1H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (1- (tetrahydro-2H-pyran-4-yl) -1H-indazol-5-yl) nicotinate (100 mg,0.283 mmol), sodium hydroxide (35 mg,0.85 mmol) was dissolved in a methanol/water (1.5 mL/0.5 mL) mixed solvent, and the reaction solution was stirred for 2 hours at 50 ℃. Water (2 mL) was added to the reaction system, followed by concentration. The aqueous phase was adjusted to pH 2 with 2N aqueous hydrochloric acid and solids precipitated. The solid was filtered and dried to give the title compound (38 mg, yellow solid), yield: 40%. MS (ESI) m/z 339.1[ M+H ]] + .
(5) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (tetrahydro-2H-pyran-4-yl) -1H-indazol-5-yl) nicotinamide
2-amino-5- (1- (tetrahydro-2H-pyran-4-yl) -1H-indazol-5-yl) nicotinic acid (30 mg,0.09 mmol) and 4-amino-bicyclo [ 2.2.2.2]Octyl-1-alkoxide (19 mg,0.11 mmol) was dissolved in N, N-dimethylformamide (2 mL), HATU (68 mg,0.18 mmol) and N, N-diisopropylethylamine (35 mg,0.27 mmol) were added. The mixture was stirred at room temperature overnight. Water (5 mL) was added to the reaction system, and the mixture was separated with ethyl acetate (5 mL. Times.2)Extraction, drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure to obtain crude product. Purification of the crude product by Prep-HPLC gave the title compound (16 mg, white solid), yield: 39%. MS (ESI) m/z 462.1[ M+H ] ] + . 1 H NMR(400 MHz,DMSO-d 6 )δ8.39(d,J＝2.4 Hz,1H),8.12(s,1H),8.03(d,J＝2.4 Hz,1H),7.99(s,1H),7.83-7.80(m,2H),7.70(dd,J＝8.8,1.6 Hz,1H),6.88(s,2H),4.93-4.90(m,1H),4.31(s,1H),4.02(dd,J＝10.8,3.6 Hz,2H),3.58(dd,J＝11.6,10.4 Hz,2H),2.18-1.98(m,8H),1.94-1.84(m,2H),1.71-1.56(m,6H).
Example 61
Compound 127: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (tetrahydro-2H-pyran-4-yl) -2H-indazol-5-yl) nicotinamide
(1) 2-amino-5- (2- (tetrahydro-2H-pyran-4-yl) -2H-indazol-5-yl) nicotinic acid methyl ester
5-bromo-2- (tetrahydro-2H-pyran-4-yl) -2H-indazole (250 mg,0.89mmol, example 60, step 1) was dissolved in a mixed solvent of 1, 4-dioxane (3 mL) and water (0.3 mL), and (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boronic acid (210 mg,1.07 mmol), potassium carbonate (307 mg,2.23 mmol) and Pd (dppf) Cl were added 2 (66 mg,0.09 mmol). The mixture was stirred at reflux under nitrogen overnight. To the reaction system was added water (20 mL), the mixture was extracted with ethyl acetate (20 ml×2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography (dichloromethane/methanol=30/1) to give the title compound (232 mg, yellow solid), yield: 74%. MS (ESI) m/z 353.1[ M+H ]] + .
(2) 2-amino-5- (2- (tetrahydro-2H-pyran-4-yl) -2H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (2- (tetrahydro-2H-pyran-4-yl) -2H-indazol-5-yl) nicotinate (230 mg,0.65 mmol), sodium hydroxide (78 mg,1.95 mmol) was dissolved in a methanol/water (6 mL/2 mL) mixed solvent. The reaction solution was heated to 50℃and stirred for 2 hours. Water (5 mL) was added to the reaction system, and the mixture was concentrated. The aqueous phase was conditioned with 2N aqueous hydrochloric acid The pH value was 2 and solids precipitated. The solid was filtered and dried to give the title compound (194 mg, yellow solid), yield: 80%. MS (ESI) m/z 339.1[ M+H ]] + .
(3) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (tetrahydro-2H-pyran-4-yl) -2H-indazol-5-yl) nicotinamide
2-amino-5- (2- (tetrahydro-2H-pyran-4-yl) -2H-indazol-5-yl) nicotinic acid (40 mg,0.12 mmol) and 4-amino-bicyclo [ 2.2.2.2]Octyl-1-alkoxide (25 mg,0.14 mmol) was dissolved in N, N-dimethylformamide (2 mL), and HATU (91 mg,0.24 mmol) and N, N-diisopropylethylamine (46 mg,0.36 mmol) were added. The mixture was stirred at room temperature overnight. Water (5 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (5 mL. Times.2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. Purification of the crude product by Prep-HPLC gave the title compound (10 mg, white solid), yield: 18.5%. MS (ESI) m/z 462.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.46(s,1H),8.38(d,J＝2.4Hz,1H),8.02(d,J＝2.4Hz,1H),7.91(s,1H),7.82(s,1H),7.68(d,J＝8.8Hz,1H),7.58(dd,J＝8.8,1.6Hz,1H),6.87(s,2H),4.79-4.73(m,1H),4.30(s,1H),4.04-4.01(m,2H),3.57-3.50(m,2H),2.18-2.03(m,10H),1.64-1.60(m,6H).
Example 62
Compound 128: 2-amino-5- (1- (1- (4, 4-difluorocyclohexyl) pyrrolidin-3-yl) -1H-indazol-5-yl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) nicotinamide
In a similar procedure as defined in example 56, starting from tert-butyl 3- (5-bromo-1H-indazol-1-yl) pyrrolidine-1-carboxylate (80.0 mg,242umol, example 56, step 1) the title compound (3.0 mg, white solid) was obtained. MS (ESI) m/z 565.1[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.38(d,J＝2.4Hz,1H),8.09(s,1H),8.03(d,J＝2.0Hz,1H),7.97(s,1H),7.88(d,J＝8.8Hz,1H),7.79(s,1H),7.68(d,J＝8.8Hz,1H),6.87(s,2H),5.40-5.32(m,1H),4.30(s,1H),3.17-3.09(m,2H),2.96-2.73(m,4H),2.37-2.22(m,5H),2.09-2.00(m,6H),1.91-1.77(m,4H),1.69-1.56(m,6H).
Example 63
Compound 129: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (2-morpholinoethyl) -2H-indazol-5-yl) nicotinamide
(1) 4- (2- (5-bromo-2H-indazol-2-yl) ethyl) morpholine
5-bromo-2-nitrobenzaldehyde (230.0 mg,1.0 mmol) and N- (2-aminoethyl) morpholine (143.0 mg,1.1 mmol) were dissolved in isopropanol (3 mL) and the reaction was stirred at 80℃for 4 hours. Cooled to room temperature, tributylphosphine (0.74 mL) was added and stirring was continued at 80 ℃ for 12 hours. To the reaction solution was added water (40 mL), extracted with ethyl acetate (30 mL. Times.3), washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography (dichloromethane/methanol=100/1 to 50/1) to give the title compound (309.0 mg, yellow oil), yield:>99％。MS(ESI):m/z 310.1[M+H] + .
(2) 2-amino-5- (2- (2-morpholinoethyl) -2H-indazol-5-yl) nicotinic acid methyl ester
4- (2- (5-bromo-2H-indazol-2-yl) ethyl) morpholine (500.0 mg,1.6 mmol) was dissolved in a mixed solvent of 1, 4-dioxane and water (24 mL/6 mL), and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (445.0 mg,1.6 mmol) was added to the mixture, [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (117 mg,0.16 mmol) and potassium carbonate (662.0 mg,4.8 mmol). The reaction was heated to 80 ℃ under nitrogen and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=40/1) to give the title compound (314.0 mg, yellow oil), yield: 62%. MS (ESI) m/z 382.1[ M+H ] ] + .
(3) 2-amino-5- (2- (2-morpholinoethyl) -2H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (2- (2-morpholinoethyl) -2H-indazol-5-yl) nicotinate (314.0 mg,0.82 mmol) was dissolved in a mixture of methanol and water (7 mL/3 mL)To the reagent was added sodium hydroxide (98.0 mg,2.46 mmol) and the mixture was heated to 50deg.C and stirred overnight. The reaction was concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (110 mg, yellow solid), yield: 37%. MS (ESI) m/z 368.1[ M+H ]] + .
(4) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (2-morpholinoethyl) -2H-indazol-5-yl) nicotinamide
2-amino-5- (2- (2-morpholinoethyl) -2H-indazol-5-yl) nicotinic acid (110 mg,0.3 mmol) was dissolved in N, N-dimethylformamide (10 mL), followed by the addition of 4-aminobicyclo [ 2.2.2.2]Octane-1-hydrochloride (53.0 mg,0.3 mmol), HATU (228.0 mg,0.6 mmol) and N, N-diisopropylethylamine (155.0 mg,1.2 mmol) were stirred at room temperature for 3 hours. To the reaction mixture was added water (40 mL), which was extracted with ethyl acetate (30 mL. Times.3), washed with a saturated sodium chloride solution, and dried over anhydrous sodium sulfate. Purification of the residue by Prep-HPLC gave the title compound (40.2 mg, white solid), yield: 27%. MS (ESI) m/z 491.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.42(s,1H),8.38(d,J＝2.4Hz,1H),8.03(d,J＝2.0Hz,1H),7.91(s,1H),7.80(s,1H),7.66(d,J＝9.2Hz,1H),7.56(dd,J＝8.8,1.6Hz,1H),6.87(s,2H),4.55(t,J＝6.4Hz,2H),4.30(s,1H),3.53(t,J＝4.8Hz,4H),2.86(t,J＝6.4Hz,2H),2.44-2.42(m,4H),2.07-2.03(m,6H),1.64-1.60(m,6H).
Example 64
Compound 130: 2-amino-5- (1- (1- (4, 4-difluorocyclohexyl) piperidin-4-yl) -1H-indazol-5-yl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) nicotinamide
(1) 4- (5-bromo-1H-indazol-1-yl) piperidine-1-carboxylic acid tert-butyl ester
5-bromo-1H-indazole (1.00 g,5.0 mmol) was dissolved in N, N-dimethylformamide (10 mL), potassium carbonate (1.42 g,10.0 mmol), tert-butyl 4- ((methylsulfonyl) oxy) piperidine-1-carboxylate (1.70 g,6.0 mmol) was added sequentially and the mixture was reacted at 100deg.C for 12 hours. Water (100 mL) was added to the reaction mixture, followed by extraction with ethyl acetate (40 mL. Times.3),the saturated sodium bicarbonate solution was washed 3 times, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=15/1) to give the title compound (900 mg, white solid), yield: 47%. MS (ESI) m/z 324.0[ M+H-56 ]] + . 1 H NMR(400MHz,CDCl 3 )δ7.93(s,1H),7.87(s,1H),7.44(dd,J＝9.2,1.6Hz,1H)7.33(d,J＝9.2Hz,1H),4.55-4.49(m,1H),4.31-4.27(m,2H),2.98-2.92(m,2H),2.26-2.15(m,2H),2.01-1.98(m,2H),1.49(s,9H).
(2) 5-bromo-1- (piperidin-4-yl) -1H-indazole hydrochloride
Tert-butyl 4- (5-bromo-1H-indazol-1-yl) piperidine-1-carboxylate (900.0 mg,2.37 mmol) was added to 4M dioxane hydrochloride solution (9 mL) and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and used directly in the next step. MS (ESI) m/z 280.0[ M+H ]] + .
(3) 5-bromo-1- (1- (4, 4-difluorocyclohexyl) piperidin-4-yl) -1H-indazole
5-bromo-1- (piperidin-4-yl) -1H-indazole hydrochloride (300.0 mg,1.07 mmol) was dissolved in methanol (3 mL), acetic acid (400 uL), 4-difluorocyclohexanone (158.0 mg,1.07 mmol) was added in sequence, and the mixture was stirred at 50℃for two hours. Sodium cyanoborohydride (135.0 mg,2.14 mmol) was then added and the reaction was continued for 3 hours. To the reaction was added water (20 mL), the mixture was extracted with dichloromethane (20 mL x 2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (100% ethyl acetate) to give the title compound (60 mg, colorless oil), yield: 14%. MS (ESI) m/z 398.0[ M+H ] ] + .
(4) 1- (1- (4, 4-difluorocyclohexyl) piperidin-4-yl) -5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indazole
5-bromo-1- (1- (4, 4-difluorocyclohexyl) piperidin-4-yl) -1H-indazole (60 mg,0.15 mmol) was dissolved in 1, 4-dioxane (1 mL) and pinacol biborate (42 mg,0.16 mmol) and [1,1' -bis (diphenylphosphine) ferrocene was added sequentially]Palladium dichloride (11 mg,0.015 mmol) and potassium acetate (29 mg,0.30 mmol). The reaction solution was heated to 70℃under nitrogen and stirred for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (100% ethyl acetate) to give the title compound (30 mg, brown)Color oil), yield: 55%. MS (ESI) m/z 446.2[ M+H ]] + .
(5) 2-amino-5- (1- (1- (4, 4-difluorocyclohexyl) piperidin-4-yl) -1H-indazol-5-yl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) nicotinamide
1- (1- (4, 4-Difluorocyclohexyl) piperidin-4-yl) -5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indazole (30.0 mg,0.067 mmol) was dissolved in a mixed solvent of 1, 4-dioxane/water (4 mL/1 mL), and [1,1' -bis (diphenylphosphine) ferrocene was sequentially added]Palladium dichloride (5 mg,0.0067 mmol), potassium carbonate (27 mg,0.20 mmol) and intermediate 5 (22 mg,0.0067 mmol). The reaction solution was heated to 70℃under nitrogen and stirred for 5 hours. The reaction solution was filtered through celite, and the filtrate was extracted with ethyl acetate (10 ml×3), washed 3 times with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by Pre-HPLC to give the title compound (1 mg, white solid). MS (ESI) m/z 579.2[ M+H ] ] + .
Example 65
Compound 131: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (1-isopropylpiperidin-4-yl) -1H-indazol-5-yl) nicotinamide
(1) 5-bromo-1- (piperidin-4-yl) -1H-indazole
Tert-butyl 4- (5-bromo-1H-indazol-1-yl) piperidine-1-carboxylate (300 mg,0.79mmol, example 64, step 1) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (5 mL) was added and the reaction stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to give the title compound (220 mg, colorless oil). MS (ESI) m/z 282.0[ M+H ]] + .
(2) 5-bromo-1- (1-isopropylpiperidin-4-yl) -1H-indazole
5-bromo-1- (piperidin-4-yl) -1H-indazole (220 mg,0.80 mmol), 2-bromopropane (97 mg,0.80 mmol) was dissolved in acetonitrile (10 mL), potassium carbonate (220 mg,1.60 mmol) was added and the mixture was heated to 80℃and stirred overnight. Water (20 mL) was added to the reaction system, the mixture was extracted with ethyl acetate (40 mL. Times.2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered,concentrating under reduced pressure. The residue was purified by column chromatography (dichloromethane/methanol=30/1) to give the title compound (120 mg, yellow oil), yield: 47%. MS (ESI) m/z 322.0[ M+H ]] + .
(3) 2-amino-5- (1- (1-isopropylpiperidin-4-yl) -1H-indazol-5-yl) nicotinic acid methyl ester
Methyl 5-bromo-1- (1-isopropylpiperidin-4-yl) -1H-indazole (120 mg,0.37 mmol), 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinic acid (104 mg,0.37 mmol), pdCl 2 (dppf) (33 mg,0.04 mmol) and potassium carbonate (102 mg,0.74 mmol) were dissolved in a mixed solvent of 1, 4-dioxane/water (10 mL/2 mL). The mixture was stirred overnight at 100℃under nitrogen. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=30/1) to give the title compound (70 mg, yellow oil), yield: 48%. MS (ESI) m/z 394.1[ M+H ]] + .
(4) 2-amino-5- (1- (1-isopropylpiperidin-4-yl) -1H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (1- (1-isopropylpiperidin-4-yl) -1H-indazol-5-yl) nicotinate (70 mg,0.18 mmol) was dissolved in a mixed solvent of methanol/water (1 mL/1 mL), and sodium hydroxide (22 mg,0.55 mmol) was added. The reaction system was stirred at 50℃for 2 hours. The reaction solution was adjusted to pH 4 with 1N hydrochloric acid and concentrated under reduced pressure. The residue was purified by reverse phase column to give the title compound (50 mg, pale yellow solid), yield: 75%. MS (ESI) m/z 380.1[ M+H ]] + .
(5) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (1-isopropylpiperidin-4-yl) -1H-indazol-5-yl) nicotinamide
2-amino-5- (1- (1-isopropylpiperidin-4-yl) -1H-indazol-5-yl) nicotinic acid (50 mg,0.14 mmol), 4-amino-bicyclo [2.2.2]Octane-1-hydrochloride (29 mg,0.16 mmol) was dissolved in N, N-dimethylformamide (1 mL), HATU (107 mg,0.28 mmol) and N, N-diisopropylethylamine (55 mg,0.42 mmol) were added, and the mixture was stirred at room temperature overnight. The reaction was purified directly by Pre-HPLC to give the title compound (2.2 mg, white solid), yield: 3.1%. MS (ESI) m/z 503.1[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.40(d,J＝2.0Hz,1H),8.11(s,1H),8.05(d,J＝2.0Hz,1H),7.99(s,1H),7.82-7.78(m,2H),7.71-7.69(m,1H),6.89(s,2H),4.64-4.57(m,1H),4.31(s,1H),2.94(d,J＝11.6Hz,2H),2.82-2.79(m,1H),2.43-2.37(m,2H),2.12-2.03(m,8H),1.96-1.92(m,2H),1.66-1.62(m,6H),1.29(d,J＝6.8Hz,6H).
Example 66
Compound 132: 2-amino-5- (2- (2- (4, 4-difluoropiperidin-1-yl) ethyl) -2H-indazol-5-yl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) nicotinamide
(1) 2- (5-bromo-2H-indazol-2-yl) ethan-1-ol
To a solution of 5-bromo-1H-indazole (4 g,20 mmol) in N, N-dimethylformamide (100 mL) was added sodium hydride (960 mg,24mmol,60% purity) under ice-bath. After the mixture was stirred for 1 hour at room temperature, 1, 3-dioxolan-2-one (7.16 g,80 mmol) was added thereto, and the mixture was stirred for 3 hours at 80 ℃. The reaction mixture was poured into water (500 mL), extracted with ethyl acetate (2×200 mL), and the organic phases were combined, washed with saturated brine (2×200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=1/1) to give the title compound (1.2 g, white solid), yield: 25%. MS (ESI) m/z 241.0[ M+H ]] + .
(2) 2- (5-bromo-2H-indazol-2-yl) methanesulfonic acid ethyl ester
2- (5-bromo-2H-indazol-2-yl) ethan-1-ol (964 mg,4 mmol) was dissolved in anhydrous dichloromethane (20 mL), and triethylamine (1.21 g,13 mmol) was added. After the mixture was cooled to 0 ℃, methanesulfonyl chloride (687 mg,6 mmol) was added dropwise, and the mixture was stirred at room temperature for 1 hour. The reaction solution was poured into water (50 mL), the aqueous phase was extracted with dichloromethane (20 mL), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title compound (1.27 g, colorless oil), yield: 98%. MS (ESI) m/z 319.0[ M+H ] ] + .
(3) 5-bromo-2- (2- (4, 4-difluoropiperidin-1-yl) ethyl) -2H-indazole
Ethyl 2- (5-bromo-2H-indazol-2-yl) methanesulfonate (1.26 g,3.95 mmol)To N, N-dimethylformamide (20 mL) was added 4, 4-difluoropiperidine hydrochloride (934 mg,5.9 mmol) and triethylamine (2 g,19.7 mmol). The reaction solution was heated to 80℃under nitrogen and stirred for 18 hours. The reaction mixture was poured into water (150 mL), extracted with ethyl acetate (2×75 mL), and the organic phases were combined, washed with saturated brine (2×100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give the title compound (260 mg, orange solid), yield: 19.2%. MS (ESI) m/z 344.0[ M+H ]] + .
(4) 2-amino-5- (2- (2- (4, 4-difluoropiperidin-1-yl) ethyl) -2H-indazol-5-yl) nicotinic acid methyl ester
5-bromo-2- (2- (4, 4-difluoropiperidin-1-yl) ethyl) -2H-indazole (103 mg,0.3 mmol) was dissolved in 1, 4-dioxane (5 mL) and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinic acid (100 mg,0.36 mmol) was added, [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (22 mg,0.03 mmol), potassium carbonate (124 mg,0.9 mmol) and water (1 mL) and the mixture was stirred at 80℃for 18 hours under nitrogen. The reaction was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=40/1) to give the title compound (80 mg, pale yellow oil), yield: 64%. MS (ESI) m/z 416.1[ M+H ] ] + .
(5) 2-amino-5- (2- (2- (4, 4-difluoropiperidin-1-yl) ethyl) -2H-indazol-5-yl) nicotinic acid
Methyl 2-amino-5- (2- (2- (4, 4-difluoropiperidin-1-yl) ethyl) -2H-indazol-5-yl) nicotinate (80 mg,0.19 mmol) was dissolved in methanol (10 mL), lithium hydroxide monohydrate (23 mg,0.57 mmol) and water (5 mL) were added and reacted at room temperature for 18 hours. The reaction was concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (40 mg, white solid), yield: 52%. MS (ESI) m/z 402.1[ M+H ]] + .
(6) 2-amino-5- (2- (2- (4, 4-difluoropiperidin-1-yl) ethyl) -2H-indazol-5-yl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) nicotinamide
2-amino-5- (2- (2- (4, 4-difluoropiperidin-1-yl) ethyl) -2H-indazol-5-yl) nicotinic acid (40 mg,0.1 mmol) was dissolved in N, N-dimethylformamide (2 mL), HATU (76 mg,0.2 mmol) and N, N-diisopropylethylamine (65 mg) were added0.5 mmol). The mixture was stirred at room temperature for 10 min and 4-aminobicyclo [ 2.2.2.2 was added]Octane-1-diol hydrochloride (21 mg,0.12 mmol) and stirring at room temperature was continued for 3 hours. The reaction was filtered and purified by Prep-HPLC to give the title compound (23.4 mg, white solid), yield: 44.6%. MS (ESI) m/z 525.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.42(s,1H),8.39(d,J＝2.0Hz,1H),8.04(d,J＝2.4Hz,1H),7.92(s,1H),7.81(s,1H),7.67(d,J＝9.2Hz,1H),7.56(dd,J＝8.8,1.2Hz,1H),6.87(s,2H),4.55(t,J＝6.4Hz,2H),4.32(s,1H),2.95(t,J＝6.4Hz,2H),2.58(t,J＝5.2Hz,4H),2.08-2.04(m,6H),1.96-1.86(m,4H),1.65-1.61(m,6H).
Example 67
Compound 133: 2-amino-N- (tetrahydro-2H-pyran-4-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinamide
(1) 2- (5-bromo-2, 3-dihydro-1H-inden-1-ylidene) acetic acid methyl ester
To a solution of methyl 2- (dimethoxyphosphono) acetate (9.46 g,52 mmol) in N, N-dimethylformamide (80 mL) was added potassium tert-butoxide (5.82 g,52 mmol), and after stirring at room temperature for 2 hours, 5-bromo-2, 3-dihydro-1H-inden-1-one (8.44 g,40 mmol) was added in portions and reacted at room temperature under nitrogen for 18 hours. The reaction mixture was poured into water (500 mL), extracted with ethyl acetate (2×150 mL), and the organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate=30/1) to give the title compound (4.38 g, yellow oil), yield: 41%. MS (ESI) m/z 267.0[ M+H ]] + .
(2) 2- (5-bromo-1- (nitromethyl) -2, 3-dihydro-1H-inden-1-yl) acetic acid methyl ester
To a solution of methyl 2- (5-bromo-2, 3-dihydro-1H-inden-1-ylidene) acetate (4.38 g,16.4 mmol) in dimethyl sulfoxide (40 mL) was added cesium carbonate (4.27 g,13.1 mmol), nitromethane (3 g,49.2 mmol) was added dropwise, and the reaction mixture was heated to 70℃under nitrogen atmosphere and stirred for 18 hours. The reaction mixture was poured into water (300 mL), extracted with ethyl acetate (2×100 mL), and the organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate=40/1) to give the title compound (1.86 g, brown oil), yield: 32%.
(3) 5-bromo-2, 3-dihydrospiro [ indene-1, 3 '-pyrrolidin ] -5' -one
To a solution of methyl 2- (5-bromo-1- (nitromethyl) -2, 3-dihydro-1H-inden-1-yl) acetate (1.86 g,5.2 mmol) in methanol (30 mL) was added water (10 mL), ammonium chloride (835 mg,15.6 mmol) and reduced iron powder (870 mg,15.6 mmol). The reaction solution was stirred at 80℃for 18 hours under nitrogen. The reaction solution was filtered through celite, the filter cake was washed with methanol (20 mL), and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=1/2) to give the title compound (1.4 g, white solid) in 78% yield. MS (ESI) m/z 266.0[ M+H ]] + .
(4) 5-bromo-2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine ]
To 5-bromo-2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine]To a solution of 5' -ketone (800 mg,3.0 mmol) in anhydrous tetrahydrofuran (20 mL) was added borane tetrahydrofuran solution (15 mL, 1M). The reaction mixture was heated to 80℃and stirred for 16 hours, cooled to room temperature, then hydrochloric acid (20 mL, 2M) was added, and the mixture was heated to 100℃and stirred for 6 hours. To the reaction solution was slowly added sodium bicarbonate to adjust pH to greater than 10, diluted with water (100 mL), extracted with dichloromethane (2 x 75 mL), the organic phases combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (756 mg, white oil). MS (ESI) m/z 252.0[ M+H ] ] + .
(5) 5-bromo-1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine ]
To a solution of 5-bromo-2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine ] (756 mg,3 mmol) in methanol (15 mL) was added tetrahydro-4H-pyran-4-one (600 mg,6 mmol) and acetic acid (1.5 mL). The mixture was heated to 50℃and stirred for 3 hours, then cooled to room temperature, sodium cyanoborohydride (540 mg,9 mmol) was added in portions and reacted at room temperature for 2 hours.
The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=30/1) to giveThe title compound (580 mg, yellow oil), yield: 57.5%. MS (ESI) m/z 336.1[ M+H ]] + .
(6) 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinic acid methyl ester
To 5-bromo-1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine]To a solution of (570 mg,1.7 mmol) 1, 4-dioxane (20 mL) was added methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (473 mg,1.7 mmol), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (124 mg,0.17 mmol), potassium carbonate (704 mg,5.1 mmol) and water (2 mL). The reaction solution was heated to 80℃under nitrogen and stirred for 18 hours. The reaction was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=30/1) to give a crude product, which was further purified by Prep-HPLC to give the title compound (210 mg, pale yellow oil), yield: 30%. MS (ESI) m/z 408.2[ M+H ] ] + .
(7) 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinic acid
To 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine]To a solution of methyl-5-yl-nicotinate (210 mg,0.52 mmol) in methanol (5 mL) was added sodium hydroxide (62 mg,1.54 mmol) and water (5 mL) and the mixture was reacted at room temperature for 18 hours. The reaction was concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (117 mg, white solid), yield: 57.6%. MS (ESI) m/z 394.1[ M+H ]] + .
(8) 2-amino-N- (tetrahydro-2H-pyran-4-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinamide
To 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine]To a solution of 5-yl) nicotinic acid (19.6 mg,0.05 mmol) in N, N-dimethylformamide (2 mL) were added HATU (38 mg,0.1 mmol) and N, N-diisopropylethylamine (38.7 mg,0.3 mmol), and after stirring at room temperature for 10 minutes, tetrahydro-2H-pyran-4-amine (10 mg,0.1 mmol) was added and the reaction was allowed to proceed at room temperature for 3 hours. The reaction was filtered and purified by Prep-HPLC to give the title compound (13.2 mg, white solid), yield: 54%. MS (ESI) m/z 477.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.38(d,J＝7.6Hz,1H),8.36(d,J＝2.0Hz,1H),8.13(d,J＝2.4Hz,1H),7.47-7.45(m,2H),7.34(d,J＝8.0Hz,1H),7.10(s,2H),4.05-3.97(m,1H),3.91-3.84(m,4H),3.42-3.36(m,3H),3.29-3.27(m,1H),2.90(t,J＝7.2Hz,2H),2.80-2.71(m,3H),2.63-2.61(m,1H),2.33-2.29(m,1H),2.17-1.91(m,4H),1.80-1.76(m,4H),1.63-1.53(m,2H),1.43-1.37(m,2H).
Example 68
Compound 134: 2-amino-N- (trans-4-hydroxycyclohexyl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinamide
Reference example 67 procedure 8 was followed using 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine)]-5-yl) nicotinic acid (19.6 mg,0.05 mmol) and trans-4-aminocyclohexane-1-ol (8.6 mg,0.075 mmol) as starting material gave the title compound (12.3 mg, white solid), yield: 49%. MS (ESI) m/z 491.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.35(d,J＝2.0Hz,1H),8.26(d,J＝7.6Hz,1H),8.09(d,J＝2.0Hz,1H),7.46-7.44(m,2H),7.33(d,J＝8.0Hz,1H),7.08(s,2H),4.57(d,J＝4.4Hz,1H),3.87-3.83(m,2H),3.75-3.71(m,1H),3.43-3.29(m,2H),2.89(t,J＝7.2Hz,2H),2.77(t,J＝7.2Hz,2H),2.72-2.59(m,2H),2.29-2.27(m,1H),2.17-1.76(m,10H),1.48-1.20(m,7H).
Example 69
Compound 135: 2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinamide
Reference example 67 procedure 8 was followed using 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine)]-5-yl) nicotinic acid (19.6 mg,0.05 mmol) and ((2S, 5R) -5-aminotetralin-2H-pyran-2-yl) methanolic hydrochloric acidStarting from salt (12.5 mg,0.075 mmol) the title compound (12.8 mg, white solid) was obtained in the yield: 50%. MS (ESI) m/z 507.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.37(d,J＝2.4Hz,1H),8.26(d,J＝7.6Hz,1H),8.11(d,J＝2.4Hz,1H),7.46-7.43(m,2H),7.34(d,J＝8.0Hz,1H),7.10(s,2H),4.64(t,J＝5.6Hz,1H),3.92-3.83(m,4H),3.43-3.38(m,2H),3.30-3.11(m,4H),2.89(t,J＝6.8Hz,2H),2.77(t,J＝6.8Hz,2H),2.72-2.60(m,2H),2.33-2.27(m,1H),2.17-1.90(m,5H),1.79-1.72(m,3H),1.67-1.55(m,1H),1.48-1.24(m,3H).
Example 70
Compound 136: 2-amino-N- ((1 r,4 r) -4-hydroxy-4-methylcyclohexyl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinamide
Reference example 67 procedure 8 was followed using 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ indene-1, 3' -pyrrolidine) ]-5-yl) nicotinic acid and (1 r,4 r) -4-amino-1-methylcyclohex-1-ol to give the title compound (16.9 mg, white solid). MS (ESI) m/z 505.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.34(d,J＝2.4Hz,1H),8.24(d,J＝7.6Hz,1H),8.08(d,J＝2.4Hz,1H),7.46-7.44(m,2H),7.34(d,J＝8.4Hz,1H),7.06(s,2H),4.33(s,1H),3.87-3.80(m,3H),3.32-3.29(m,2H),2.89(t,J＝7.2Hz,2H),2.78(t,J＝6.4Hz,2H),2.74-2.61(m,2H),2.35-2.25(m,1H),2.17-1.98(m,2H),1.96-1.90(m,2H),1.79-1.73(m,4H),1.61-1.59(m,2H),1.51-1.38(m,6H),1.16(s,3H).
Example 71
Compound 137: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ inden-1, 4' -piperidin ] -5-yl) nicotinamide
(1) 5-Bromopiro [ indene-1, 4 '-piperidine ] -1' -carboxylic acid tert-butyl ester
5-bromo-1H-indene (2.0 g,10.2 mmol) was dissolved in tetrahydrofuran (20 mL), a 1M tetrahydrofuran solution of lithium bis (trimethylsilylamide) (20.4 mL,20.4 mmol) was added dropwise at 0deg.C, and stirring was performed at 0deg.C for 1 hour. Water (20 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (40 mL. Times.2) in portions, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=20/1) to give the title compound and its isomer 6-bromospiro [ indene-1, 4' -piperidine]Mixtures of tert-butyl 1' -carboxylate (2.4 g, yellow oil), yield: 65%. MS (ESI) m/z 308.0[ M+H-56 ]] + .
(2) 5-Bromopiro [ indene-1, 4' -piperidine ]
5-Bromopiro [ indene-1, 4' -piperidine]-1 '-carboxylic acid tert-butyl ester and its isomer 6-bromospiro [ indene-1, 4' -piperidine ]A mixture of tert-butyl 1' -carboxylate (500 mg,1.4 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (5 mL) was added dropwise and stirred at room temperature for 1 hour. The reaction mixture was dried by spinning, saturated aqueous sodium bicarbonate (10 mL) was added, the mixture was extracted with dichloromethane (10 mL. Times.2) and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound and its isomer 6-bromospiro [ indene-1, 4' -piperidine](362 mg, yellow oil) in 97.9% yield. MS (ESI) m/z 264.0[ M+H ]] + .
(3) 5-bromo-1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine ]
5-Bromopiro [ indene-1, 4' -piperidine]And the isomer 6-bromospiro [ indene-1, 4' -piperidine]To the mixture (362 mg,1.4 mmol) of tetrahydro-4H-pyran-4-one (165 mg,1.7 mmol) was dissolved in methanol (5 mL), and the mixture was stirred at room temperature for half an hour, sodium cyanoborohydride (170 mg,2.7 mmol) was added, and the reaction was stirred at room temperature overnight. To the reaction solution was added water (20 mL), and the mixture was extracted with ethyl acetate (40 ml×2) in portions, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (100% ethyl acetate) to give the title compound and its isomer 6-bromo-1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine](290 mg, yellow oil), yield: 60%. MS (ESI) m/z 348.1[ M+H ] ] + .
(4) 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ inden-1, 4' -piperidin ] -5-yl) nicotinic acid methyl ester
5-bromo-1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine]And the isomer 6-bromo-1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine](640 mg,1.8 mmol), methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (516 mg,1.8 mmol), pdCl 2 (dppf) (147 mg,0.2 mmol) and potassium carbonate (497 mg,3.6 mmol) were dissolved in a mixed solvent of 1, 4-dioxane/water (10 mL/2 mL). The temperature was raised to 100℃under nitrogen and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=30/1) to give the title compound and its isomer 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine]Mixture of methyl 6-yl nicotinate (400 mg, yellow solid), yield: 52%. MS (ESI) m/z 420.1[ M+H ]] + .
(5) 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ inden-1, 4' -piperidin ] -5-yl) nicotinic acid
2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine)]-5-yl) nicotinic acid methyl ester and its isomer 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine)]A mixture of methyl-6-yl-nicotinate (200 mg,0.5 mmol) was dissolved in a mixed solvent of methanol/water (5 mL/5 mL), sodium hydroxide (57 mg,1.4 mmol) was added, and the reaction was stirred at 50℃for 2 hours. The reaction solution was dried by spin-drying to remove methanol, the pH was adjusted to 3-4 with dilute hydrochloric acid, and concentrated under reduced pressure. The residue was purified by reverse phase column to give the title compound and its isomer 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine) ]Mixtures of 6-yl nicotinic acid (170 mg, yellow solid), yield: 88%. MS (ESI) m/z 406.1[ M+H ]] + .
(6) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ inden-1, 4' -piperidin ] -5-yl) nicotinamide
2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine)]-5-yl) nicotinic acid and its isomer 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ indene-1, 4' -piperidine)]Mixtures of 6-yl nicotinic acid (170 mg,0.42 mmol), 4-amino-bicyclo [2.2.2]Octane-1-hydrochloride (75 mg,0.42 mmol) was dissolvedTo N, N-dimethylformamide (5 mL) was added HATU (319 mg,0.84 mmol) and N, N-diisopropylethylamine (155 mg,1.20 mmol), and the mixture was stirred overnight at room temperature. Water (20 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (20 mL. Times.2) in portions, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase column to give 140mg of crude mixture. 40mg of the crude mixture was purified by Pre-HPLC to give the title compound (8.2 mg, white solid) and another isomer. MS (ESI) m/z 529.2[ M+H ]] + .1HNMR(400MHz,DMSO-d 6 )δ8.33(d,J＝2.4Hz,1H),8.00(d,J＝2.4Hz,1H),7.74(s,1H),7.56(d,J＝1.2Hz,1H),7.50-7.41(m,2H),6.99(d,J＝6.0Hz,1H),6.90(s,2H),6.83(d,J＝5.6Hz,1H),4.29(s,1H),3.94-3.90(m,2H),3.31-3.29(m,2H),2.99-2.96(m,2H),2.56-2.54(m,3H),2.11-2.02(m,8H),1.79-1.76(m,2H),1.63-1.59(m,6H),1.55-1.45(m,2H),1.26-1.23(m,2H).
Example 72
Compound 138: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) spiro [ inden-1, 4' -piperidin ] -6-yl) nicotinamide
Referring to example 71, step 6, the 40mg crude mixture was purified by Pre-HPLC to afford the other isomer, the title compound (16 mg, white solid). MS (ESI) m/z 529.2[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.38(d,J＝2.4Hz,1H),7.98(d,J＝2.4Hz,1H),7.79(s,1H),7.69(s,1H),7.50-7.47(m,2H),6.96(d,J＝5.6Hz,1H),6.87(s,2H),6.80(d,J＝5.6Hz,1H),4.30(s,1H),3.94-3.90(m,2H),3.31-3.28(m,2H),3.01-2.98(m,2H),2.53-2.48(m,3H),2.21-2.15(m,2H),2.07-2.03(m,6H),1.79-1.77(m,2H),1.64-1.60(m,6H),1.55-1.46(m,2H),1.26-1.23(m,2H).
Example 73
Compound 139: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 4' -piperidin ] -5-yl) nicotinamide
The crude mixture (100 mg,0.19 mmol) obtained in step 6 of example 71 was dissolved in methanol (5 mL), palladium on carbon (10 mg) was added, and the reaction mixture was stirred overnight at room temperature under hydrogen. Filtration through celite, concentration under reduced pressure, and purification of the residue by Pre-HPLC afforded the title compound (21.5 mg, white solid) and another isomer. MS (ESI) m/z 531.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.30(d,J＝2.4Hz,1H),7.96(d,J＝2.8Hz,1H),7.74(s,1H),7.43-7.40(m,2H),7.25(d,J＝8.0Hz,1H),6.88(s,2H),4.31(s,1H),3.91-3.88(m,2H),3.31-3.26(m,2H),2.90-2.84(m,4H),2.47-2.41(m,1H),2.29(t,J＝11.2Hz,2H),2.05-2.01(m,6H),1.97(t,J＝7.6Hz,2H),1.83-1.70(m,4H),1.63-1.59(m,6H),1.50-1.41(m,4H).
Example 74
Compound 140: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 4' -piperidin ] -6-yl) nicotinamide
Referring to example 73, this step was purified by Pre-HPLC to give the other isomer, the title compound (12.5 mg, white solid). MS (ESI) m/z 531.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.33(d,J＝2.4Hz,1H),7.92(d,J＝2.4Hz,1H),7.78(s,1H),7.44(s,1H),7.40-7.38(m,1H),7.24(d,J＝7.6Hz,1H),6.85(s,2H),4.29(s,1H),3.92-3.88(m,2H),3.29-3.26(m,2H),2.89-2.83(m,4H),2.47-2.46(m,1H),2.30(t,J＝12.0Hz,2H),2.06-2.02(m,6H),1.97(t,J＝7.6Hz,2H),1.93-1.86(m,2H),1.74-1.71(m,2H),1.64-1.60(m,6H),1.49-1.43(m,4H).
Example 75
Compound 141: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -3H-spiro [ isobenzofuran-1, 4' -piperidin ] -5-yl) nicotinamide
(1) 5-bromo-3-oxo-3H-spiro [ isobenzofuran-1, 4 '-piperidine ] -1' -carboxylic acid tert-butyl ester
Methyl 5-bromo-2-iodobenzoate (5.0 g,14.70 mmol) and tert-butyl 4-oxopiperidine-1-carboxylate (3.2 g,16.20 mmol) were dissolved in tetrahydrofuran (50 mL). Cooled to-78 ℃, and a solution of isopropyl magnesium chloride lithium chloride complex (1.3 m,12.5ml,16.20 mmol) in tetrahydrofuran was added dropwise to the reaction solution. The reaction was stirred overnight at room temperature under nitrogen. To the reaction solution was added water (20 mL), which was filtered through celite, the filtrate was extracted with ethyl acetate (20 mL. Times.2), and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give the title compound (3.9 g, white solid), yield: 70%. MS (ESI) m/z 326.0[ M+H-56 ] ] + .
(2) 4- (4-bromo-2- (hydroxymethyl) phenyl) -4-hydroxypiperidine-1-carboxylic acid tert-butyl ester
5-bromo-3-oxo-3H-spiro [ isobenzofuran-1, 4' -piperidine]Tert-butyl 1' -carboxylate (3.8 g,9.97 mmol) was dissolved in tetrahydrofuran (40 mL) and a solution of lithium borohydride (2M, 10mL,19.94 mmol) in tetrahydrofuran was added dropwise at 0deg.C. The reaction was stirred overnight at room temperature under nitrogen. Water (20 mL) was added to the reaction system, the mixture was extracted with ethyl acetate (20 mL. Times.2), and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=3/1) to give the title compound (3.8 g, colorless oil), yield: 99%. MS (ESI) m/z 312.0[ M+H-56-H 2 O] + .
(3) 5-bromo-3H-spiro [ isobenzofuran-1, 4 '-piperidine ] -1' -carboxylic acid tert-butyl ester
4- (4-bromo-2- (hydroxymethyl) phenyl) -4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (2.0 g,5.2 mmol) was dissolved in tetrahydrofuran (20 mL) and triethylamine (2.6 g,26.0 mmol) and p-toluenesulfonic anhydride (2.2 g,6.8 mmol) were added at 0deg.C. The reaction was stirred at room temperature under nitrogen overnight. Water (20 mL) was added to the reaction system, extracted with ethyl acetate (20 mL. Times.2), and the organic phase was usedSaturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=15/1) to give the title compound (1.2 g, colorless oil), yield: 63%. MS (ESI): m/z 268.0[ M+H-100 ] ] + .
(4) 5-bromo-3H-spiro [ isobenzofuran-1, 4' -piperidine ]
5-bromo-3H-spiro [ isobenzofuran-1, 4' -piperidine]Tert-butyl 1' -carboxylate (300 mg,0.82 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (5 mL) was added dropwise, and the reaction was stirred at room temperature for 1 hour. The reaction was dried by spinning, saturated aqueous sodium hydrogencarbonate (10 mL) was added to the reaction system, extracted by liquid-split extraction with methylene chloride (10 ml×2), dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound (210 mg, white solid), yield: 96%. MS (ESI): m/z 268.0[ M+H ]] + .
(5) 5-bromo-1 '- (tetrahydro-2H-pyran-4-yl) -3H-spiro [ isobenzofuran-1, 4' -piperidine ]
5-bromo-3H-spiro [ isobenzofuran-1, 4' -piperidine](210 mg,0.79 mmol) tetrahydro-4H-pyran-4-one (157 mg,1.57 mmol) is dissolved in methanol (5 mL) and stirred at room temperature for half an hour, sodium cyanoborohydride (151 mg,2.37 mmol) is added and stirring is continued at room temperature overnight. Methanol was dried by spinning, water (20 mL) was added to the reaction system, extracted by liquid-separated extraction with ethyl acetate (40 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography (100% ethyl acetate) to give the title compound (120 mg, white solid), yield: 43%. MS (ESI) M/z352.1[ M+H ]] + .
(6) 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -3H-spiro [ isobenzofuran-1, 4' -piperidin ] -5-yl) nicotinic acid methyl ester
5-bromo-1 '- (tetrahydro-2H-pyran-4-yl) -3H-spiro [ isobenzofuran-1, 4' -piperidine](120 mg,0.34 mmol) methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (95 mg,0.34 mmol), pdCl 2 (dppf) (25 mg,0.03 mmol) and potassium carbonate (94 mg,0.68 mmol) were dissolved in a mixed solvent of 1, 4-dioxane/water (10 mL/2 mL). The reaction solution was stirred overnight at 100℃under nitrogen. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=20/1) to give the title compound (100)mg, yellow solid), yield: 69%. MS (ESI) m/z 424.2[ M+H ]] + .
(7) 2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -3H-spiro [ isobenzofuran-1, 4' -piperidin ] -5-yl) nicotinic acid
2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -3H-spiro [ isobenzofuran-1, 4' -piperidine]5-yl) nicotinic acid methyl ester (100 mg,0.24 mmol) was dissolved in a mixed solvent of methanol/water (5 mL/5 mL), sodium hydroxide (29 mg,0.72 mmol) was added, and the reaction system was stirred at 50℃for 2 hours. The reaction solution is dried by rotating methanol, the pH value is regulated to 3-4 by dilute hydrochloric acid, and the solution is concentrated under reduced pressure. The residue was purified by reverse phase column to give the title compound (50 mg, white solid), yield: 52%. MS (ESI) m/z 410.1[ M+H ] ] + .
(8) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -3H-spiro [ isobenzofuran-1, 4' -piperidin ] -5-yl) nicotinamide
2-amino-5- (1 '- (tetrahydro-2H-pyran-4-yl) -3H-spiro [ isobenzofuran-1, 4' -piperidine]-5-yl) nicotinic acid (30 mg,0.07 mmol), 4-amino-bicyclo [2.2.2]Octane-1-hydrochloride (13 mg,0.07 mmol) was dissolved in N, N-dimethylformamide (5 mL), HATU (42 mg,0.11 mmol) and N, N-diisopropylethylamine (18 mg,0.14 mmol) were added, and the reaction was stirred at room temperature overnight. The reaction was purified directly by Pre-HPLC to give the title compound (14 mg, white solid), yield: 36%. MS (ESI) m/z 533.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.34(d,J＝2.0Hz,1H),7.99(d,J＝2.0Hz,1H),7.74(s,1H),7.54-7.52(m,2H),7.33(d,J＝7.6Hz,1H),6.92(s,2H),5.01(s,2H),4.29(s,1H),3.93-3.89(m,2H),3.30-3.27(m,2H),2.82-2.80(m,2H),2.51-2.50(m,3H),2.06-2.02(m,6H),1.93-1.86(m,2H),1.74-1.60(m,10H),1.53-1.44(m,2H).
Example 76
Compound 142: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '-isopropyl-3H-spiro [ isobenzofuran-1, 4' -piperidin ] -5-yl) nicotinamide
(1) 5-bromo-1 '-isopropyl-3H-spiro [ isobenzofuran-1, 4' -piperidine ]
5-bromo-3H-spiro [ isobenzofuran-1, 4' -piperidine](218 mg,0.80mmol, example 75, step 4) 2-bromopropane (200 mg,1.60 mmol) was dissolved in acetonitrile (10 mL) and potassium carbonate (331 mg,2.40 mmol) was added and the mixture heated to 80℃and stirred overnight. Water (20 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (40 mL. Times.2) in a liquid-separated manner, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (100% ethyl acetate) to give the title compound (150 mg, colorless oil), yield: 60%. MS (ESI) m/z 310.1[ M+H ] ] + .
(2) 2-amino-5- (1 '-isopropyl-3H-spiro [ isobenzofuran-1, 4' -piperidin ] -5-yl) nicotinic acid methyl ester
5-bromo-1 '-isopropyl-3H-spiro [ isobenzofuran-1, 4' -piperidine](150 mg,0.49 mmol) methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (135 mg,0.49 mmol), pdCl 2 (dppf) (41 mg,0.05 mmol), potassium carbonate (138 mg,1.00 mmol) was dissolved in a mixed solvent of 1, 4-dioxane/water (10 mL/2 mL). The mixture was stirred overnight at 100℃under nitrogen. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=20/1) to give the title compound (160 mg, white solid), yield: 86%. MS (ESI) m/z 382.2[ M+H ]] + .
(3) 2-amino-5- (1 '-isopropyl-3H-spiro [ isobenzofuran-1, 4' -piperidin ] -5-yl) nicotinic acid
2-amino-5- (1 '-isopropyl-3H-spiro [ isobenzofuran-1, 4' -piperidine)]5-yl) methyl nicotinate (160 mg,0.42 mmol) was dissolved in a mixed solvent of methanol/water (5 mL/5 mL) and sodium hydroxide (50 mg,1.26 mmol) was added. The mixture was stirred at 50℃for 2 hours. The reaction was dried with screw to adjust the pH to 3-4 with dilute hydrochloric acid, concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (78 mg, white solid), yield: 50%. MS (ESI) m/z 368.2[ M+H ] ] + .
(4) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '-isopropyl-3H-spiro [ isobenzofuran-1, 4' -piperidin ] -5-yl) nicotinamide
2-amino-5- (1 '-isopropyl-3H-spiro [ isobenzofuran-1, 4' -piperidine)]-5-yl) nicotinic acid (40 mg,0.11 mmol), 4-amino-bicyclo [2.2.2]Octane-1-hydrochloride (19 mg,0.11 mmol) was dissolved in N, N-dimethylformamide (5 mL), HATU (63 mg,0.17 mmol) and N, N-diisopropylethylamine (28 mg,0.22 mmol) were added, and stirred at room temperature overnight. The reaction was purified by Pre-HPLC to give the title compound (27 mg, white solid), yield: 51%. MS (ESI) m/z 491.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.34(d,J＝2.4Hz,1H),7.99(d,J＝2.4Hz,1H),7.74(s,1H),7.54-7.52(m,2H),7.32(d,J＝7.6Hz,1H),6.92(s,2H),5.01(s,2H),4.29(s,1H),2.77-2.69(m,3H),2.53-2.48(m,2H),2.06-2.02(m,6H),1.92-1.85(m,2H),1.66-1.60(m,8H),1.02(d,J＝6.8Hz,6H).
Example 77
Compound 143: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinolin-6-yl) nicotinamide
(1) 6-bromo-2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinoline
6-bromo-1, 2,3, 4-tetrahydroisoquinoline (500 mg,2.4 mmol) and tetrahydro-4H-pyran-4-one (237 mg,2.4 mmol) are dissolved in methanol (10 mL), stirred at room temperature for half an hour, sodium cyanoborohydride (296.1 mg,4.7 mmol) is added and the reaction is stirred at room temperature overnight. Water (20 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (40 mL. Times.2) in portions, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=2/1) to give the title compound (550 mg, yellow solid), yield: 79%. MS (ESI) m/z 296.0[ M+H ] ] + .
(2) 2-amino-5- (2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinolin-6-yl) nicotinic acid methyl ester
6-bromo-2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinoline (250 mg,0.85 mmol), (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boronic acid (236 mg,0.85 mmol), pdCl 2 (dppf) (65 mg,0.08 mmol) and potassium carbonate (234 mg,1.7 mmol) were dissolved in a mixed solvent of dioxane/water (5 mL/1 mL).The temperature was raised to 100℃under nitrogen and the reaction was stirred overnight. The reaction was concentrated, and the residue was purified by column chromatography (100% ethyl acetate) to give the title compound (110 mg, yellow solid), yield: 35%. MS (ESI) m/z 368.1[ M+H ]] + .
(3) 2-amino-5- (2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinolin-6-yl) nicotinic acid
Methyl 2-amino-5- (2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinolin-6-yl) nicotinate (110 mg,0.3 mmol) was dissolved in a mixed solvent of methanol/water (5 mL/5 mL), sodium hydroxide (36 mg,0.9 mmol) was added, and the reaction solution was stirred at room temperature overnight. The reaction solution was dried by spin-drying to remove methanol, the pH was adjusted to 3-4 with dilute hydrochloric acid, and concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (100 mg, yellow solid), yield: 94%. MS (ESI) m/z 354.1[ M+H ]] + .
(4) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinolin-6-yl) nicotinamide
2-amino-5- (2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinolin-6-yl) nicotinic acid (80 mg,0.23 mmol), 4-amino-bicyclo [ 2.2.2.2]Octane-1-hydrochloride (40 mg,0.23 mmol) was dissolved in N, N-dimethylformamide (5 mL), HATU (175 mg,0.46 mmol) and N, N-diisopropylethylamine (89 mg,0.69 mmol) were added, and stirred at room temperature overnight. The reaction was purified directly by Pre-HPLC to give the title compound (16 mg, white solid), yield: 15%. MS (ESI) m/z 477.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 ) Delta 8.32 (d, j=2.4 hz, 1H), 7.96 (d, j=2.0 hz, 1H), 7.77 (s, 1H), 7.39 (d, j=8.0 hz, 1H), 7.36 (s, 1H), 7.11 (d, j=8.0 hz, 1H), 6.87 (s, 2H), 4.29 (s, 1H), 3.92 (dd, j=11.6, 2.8hz, 2H), 3.71 (s, 2H), 3.34-3.29 (m, 2H), 2.83-2.77 (m, 4H), 2.60-2.58 (m, 1H), 2.06-2.02 (m, 6H), 1.80-1.77 (m, 2H), 1.63-1.59 (m, 6H), 1.57-1.47 (m, 2H) and example 78
Compound 144: 2-amino-5- (4- (4-ethoxy-1-isopropylpiperidin-4-yl) phenyl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) nicotinamide
(1) 4- (4-bromophenyl) -4-ethoxypiperidine-1-carboxylic acid tert-butyl ester
4- (4-bromophenyl) -4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (900.0 mg,2.5 mmol) was dissolved in N, N-dimethylformamide (15 mL), and sodium hydride (91.0 mg,3.8 mmol) was added at 0deg.C and stirred for 2 hours. A solution of iodoethane (593.0 mg,3.8 mmol) in N, N-dimethylformamide (3 mL) was then added and the reaction stirred at room temperature overnight. Water (80 mL) was added to the reaction solution, which was extracted with ethyl acetate (40 mL. Times.3), washed 3 times with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane/methanol=15/1) to give the title compound (320.0 mg, white solid), yield: 33%. MS (ESI) m/z 284.0[ M+H-100 ] ] + .
(2) 4- (4-bromophenyl) -4-ethoxypiperidine hydrochloride
4- (4-bromophenyl) -4-ethoxypiperidine-1-carboxylic acid tert-butyl ester (320.0 mg,0.83 mmol) was dissolved in dichloromethane (5 mL), and a 4M dioxane solution (2 mL,8 mmol) of hydrochloric acid was added thereto, followed by stirring at room temperature for 2 hours. The reaction solution was concentrated to give the title compound, which was directly used in the next reaction. MS (ESI) m/z 284.0[ M+H ]] + .
(3) 4- (4-bromophenyl) -4-ethoxy-1-isopropylpiperidine
The crude product obtained in the previous step was dissolved in acetonitrile (10 mL), 2-bromopropane (510.0 mg,4.15 mmol) and potassium carbonate (344.0 mg,2.49 mmol) were added sequentially, and the mixture was heated to 80℃under nitrogen and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=40/1 to 20/1) to give the title compound (202 mg, white solid), yield: 75%. MS (ESI) m/z 326.1[ M+H ]] + .
(4) 2-amino-5- (4- (4-ethoxy-1-isopropylpiperidin-4-yl) phenyl) nicotinic acid methyl ester
4- (4-bromophenyl) -4-ethoxy-1-isopropyl piperidine (202 mg,0.62 mmol) was dissolved in a mixed solvent of 1, 4-dioxane/water (7 mL/1.5 mL), and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinic acid (207 mg,0.74 mmol) and [1,1' -bis (diphenylphosphine) ferrocene were sequentially added ]Palladium dichloride (45 mg,0.06 mmol) and carbonPotassium acid (257 mg,1.86 mmol). The reaction solution was heated to 80℃under nitrogen and stirred for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=50/1 to 25/1) to give the title compound (140 mg, brown oil), yield: 57%. MS (ESI) m/z 398.2[ M+H ]] + .
(5) 2-amino-5- (4- (4-ethoxy-1-isopropylpiperidin-4-yl) phenyl) nicotinic acid
Methyl 2-amino-5- (4- (4-ethoxy-1-isopropylpiperidin-4-yl) phenyl) nicotinate (100.0 mg,0.25 mmol) was dissolved in a mixed solvent of methanol/water (4 mL/1 mL) and sodium hydroxide (30.0 mg,0.75 mmol) was added. The reaction solution was stirred at 50℃for 2 hours. The reaction was concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (66 mg, yellow solid), yield: 69%. MS (ESI) m/z 384.1[ M+H ]] + .
(6) 2-amino-5- (4- (4-ethoxy-1-isopropylpiperidin-4-yl) phenyl) -N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) nicotinamide
2-amino-5- (4- (4-ethoxy-1-isopropylpiperidin-4-yl) phenyl) nicotinic acid (66.0 mg,0.17 mmol) was dissolved in N, N-dimethylformamide (3 mL) followed by the addition of 4-amino-bicyclo [ 2.2.2.2)]Octane-1-ol hydrochloride (30.0 mg,0.17 mmol), HATU (129.0 mg,0.34 mmol) and N, N-diisopropylethylamine (132.0 mg,1.02 mmol). The reaction solution was stirred at room temperature for 2 hours. The reaction was concentrated under reduced pressure, and the residue was purified by Prep-HPLC to give the title compound (17.6 mg, white solid), yield: 20%. MS (ESI) m/z 507.2[ M+H ] ] + .1HNMR(400MHz,DMSO-d 6 )δ8.36(d,J＝2.0Hz,1H),8.02(d,J＝2.4Hz,1H),7.77(s,1H),7.62-7.60(m,2H),7.46-7.44(m,2H),6.94(s,2H),4.30(s,1H),3.04(q,J＝7.2Hz,2H),2.71-2.54(m,5H),2.06-1.94(m,8H),1.87-1.80(m,2H),1.63-1.60(m,6H),1.06(t,J＝7.2Hz,3H),0.99(d,J＝6.8Hz,6H).
Example 79
Compound 145:6' -amino-5- (4-ethoxy-1-isopropylpiperidin-4-yl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) - [2,3' -bipyridine ] -5' -carboxamide
(1) 4- (6-bromopyridin-3-yl) -4-hydroxypiperidine-1-carboxylic acid tert-butyl ester
2-bromo-5-iodopyridine (4 g,14 mmol) was dissolved in tetrahydrofuran (50 mL), and n-butyllithium/n-hexane solution (2.5M, 6.4mL,16 mmol) was added dropwise at-78deg.C and stirred at-78deg.C for 2 hours. A solution of tert-butyl 4-oxopiperidine-1-carboxylate (2.8 g,14 mmol) in tetrahydrofuran (10 mL) was then added and stirring continued at-78deg.C for 1 hour. The reaction mixture was quenched by slow addition of water (80 mL), extracted with ethyl acetate (30 mL. Times.3), and the organic phase was washed with saturated sodium chloride solution (100 mL. Times.2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=5/1) to give the title compound (2.3 g, pale yellow oil), yield: 46%. MS (ESI): m/z 357.1[ M+H ]] + .
(2) 4- (6-bromopyridin-3-yl) -4-ethoxypiperidine-1-carboxylic acid tert-butyl ester
4- (6-bromopyridin-3-yl) -4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (1 g,2.8 mmol) was dissolved in N, N-dimethylformamide (20 mL), and sodium hydride (336 mg,8.4 mmol) was added thereto at 0℃and stirred for 2 hours. A solution of iodoethane (655 mg,4.2 mmol) in N, N-dimethylformamide (3 mL) was then added and the mixture was allowed to warm to room temperature and stirred overnight. To the reaction solution was added water (200 mL), extracted with ethyl acetate (100 mL. Times.3), washed 2 times with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether/ethyl acetate=5/1) to give the title compound (500.0 mg, pale yellow oil), yield: 20%. MS (ESI) m/z 385.0[ M+H ] ] + .
(3) 2-bromo-5- (4-ethoxypiperidin-4-yl) pyridine hydrochloride
4- (6-bromopyridin-3-yl) -4-ethoxypiperidine-1-carboxylic acid tert-butyl ester (500 mg,1.3 mmol) was dissolved in dichloromethane (7 mL), and a 4M dioxane solution (3 mL,12 mmol) of hydrochloric acid was added thereto, followed by stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to give the title compound, which was directly used in the next reaction. MS (ESI) m/z 285.0[ M+H ]] + .
(4) 2-chloro-5- (4-ethoxy-1-isopropylpiperidin-4-yl) pyridine
The 2-bromo-5- (4-ethoxypiperidin-4-yl) pyridine hydrochloride obtained in the previous step was dissolved in acetonitrile solution (15 mL), followed by addition of 2-bromopropane (800.0 mg,6.5 mmol) and potassium carbonate (538 mg,3.9 mmol). The reaction was heated to 80 ℃ under nitrogen and stirred overnight. To the reaction solution was added water (50 mL), extracted with ethyl acetate (30 ml×3), washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the title compound (300 mg, yellow solid), yield: 82%. MS (ESI) m/z 283.2[ M+H ]] + .
(5) 6' -amino-5- (4-ethoxy-1-isopropylpiperidin-4-yl) - [2,3' -bipyridine ] -5' -carboxylic acid methyl ester
2-chloro-5- (4-ethoxy-1-isopropylpiperidin-4-yl) pyridine (300 mg,1.05 mmol) was dissolved in a mixed solution of 1, 4-dioxane/water (4 mL/1 mL), and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinic acid (350 mg,1.26 mmol) and [1,1' -bis (diphenylphosphine) ferrocene were added sequentially ]Palladium dichloride (80 mg,0.11 mmol) and potassium carbonate (435 mg,3.15 mmol). The reaction solution was heated to 80℃under nitrogen and stirred for 18 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=10/1) to give the title compound (300 mg, black oil), yield: 71%. MS (ESI) m/z 399.2[ M+H ]] + .
(6) 6' -amino-5- (4-ethoxy-1-isopropylpiperidin-4-yl) - [2,3' -bipyridine ] -5' -carboxylic acid
6 '-amino-5- (4-ethoxy-1-isopropylpiperidin-4-yl) - [2,3' -bipyridine]Methyl 5' -carboxylate (300.0 mg,0.75 mmol) was dissolved in a mixed solvent of methanol/water (4 mL/1 mL), sodium hydroxide (91.0 mg,2.27 mmol) was added, and the system was stirred at 50℃for 12 hours. The reaction was concentrated under reduced pressure, and the residue was purified by reverse phase column to give the title compound (130 mg, yellow solid), yield: 45%. MS (ESI) m/z 385.2[ M+H ]] + .
(7) 6' -amino-5- (4-ethoxy-1-isopropylpiperidin-4-yl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) - [2,3' -bipyridine ] -5' -carboxamide
6 '-amino-5- (4-ethoxy-1-isopropylpiperidin-4-yl) - [2,3' -bipyridine]5' -Carboxylic acid (130.0 mg,0.26 mmol) was dissolved in N, N-dimethylformamide (2 mL), followed by addition of 4-amino groupBicyclo [2.2.2]Octane-1-ol hydrochloride (51 mg,0.29 mmol), HATU (198mg, 0.52 mmol) and N, N-diisopropylethylamine (101 mg,0.78 mmol) were stirred at room temperature for 1 hour. The reaction was concentrated under reduced pressure, and the residue was purified by Prep-HPLC to give the title compound (58.5 mg, pale yellow solid), yield: 44%. MS (ESI) m/z 508.2[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.76(d,J＝2.0Hz,1H),8.62(d,J＝1.6Hz,1H),8.36(d,J＝2.4Hz,1H),7.89-7.81(m,3H),7.08(s,2H),4.30(s,1H),3.07(q,J＝6.8Hz,2H),2.71-2.54(m,5H),2.07-1.99(m,8H),1.92-1.85(m,2H),1.65-1.61(m,6H),1.08(t,J＝6.8Hz,3H),1.00(d,J＝6.8Hz,6H).
Example 80
Compound 148: (R) -2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydrospiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinamide
Compound 115 (90 mg crude, example 57) was resolved by chiral column to give the title compound (16.6 mg, white solid) as a corresponding retention time of 8.022 minutes with an ee value of 100% and an absolute configuration of undetermined. MS (ESI) m/z 517.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.31(d,J＝2.4Hz,1H),7.96(d,J＝2.4Hz,1H),7.75(s,1H),7.45-7.43(m,2H),7.33(d,J＝8.4Hz,1H),6.88(s,2H),4.30(s,1H),3.88-3.85(m,2H),3.29-3.25(m,2H),2.89(t,J＝7.2Hz,2H),2.81-2.79(m,2H),2.75-2.62(m,2H),2.39-2.31(m,1H),2.17-2.10(m,1H),2.06-2.02(m,7H),1.99-1.90(m,2H),1.81-1.78(m,2H),1.64-1.60(m,6H),1.48-1.43(m,2H).
Example 81
Compound 149: (S) -2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1 '- (tetrahydro-2H-pyran-4-yl) -2, 3-dihydro-spiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinamide
Compound 115 (90 mg crude, example 57) was resolved by chiral column to give the title compound (14.7 mg, white solid) as a corresponding retention time 12.402 min, ee 99.7%, absolute configuration undefined. MS (ESI) m/z 517.1[ M+H ]] + . 1 H NMR(400M Hz,DMSO-d 6 )δ8.31(d,J＝2.4Hz,1H),7.96(d,J＝2.4Hz,1H),7.75(s,1H),7.45-7.43(m,2H),7.33(d,J＝8.4Hz,1H),6.88(s,2H),4.30(s,1H),3.88-3.85(m,2H),3.29-3.25(m,2H),2.89(t,J＝7.2Hz,2H),2.84-2.76(m,2H),2.74-2.63(m,2H),2.39-2.31(m,1H),2.17-2.10(m,1H),2.06-2.02(m,7H),1.99-1.90(m,2H),1.81-1.78(m,2H),1.64-1.60(m,6H),1.48-1.43(m,2H).
Example 82
Compound 56: 2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) -5- (4- (7- (oxetan-3-yl) -2, 7-diazaspiro [3.5] nonan-2-yl) phenyl) nicotinamide
Reference example 30 procedure of steps (2) to (4) 2- (4-bromophenyl) -7- (oxetan-3-yl) -2, 7-diazaspiro [3.5]Nonane and (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boronic acid were used as starting materials to give the title compound (1.1 mg, white solid) via similar procedure. MS (ESI) m/z 508.3[ M+H ] ] + .
Example 83
Compound 59: 2-amino-5- (4- (morpholinomethyl) phenyl) -N- (piperidin-4-yl) nicotinamide
Referring to the procedure of example 23, step (3) was performed by condensation reaction using tert-butyl 4-aminopiperidine-1-carboxylate instead of tetrahydro-2H-pyran-4-amine, deprotection with hydrochloric acid and purification by Pre-HPLC gave the title compound (11.8 mg, white solid). MS (ESI) m/z 396.4[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.40(s,1H),8.37(d,J＝7.0Hz,1H),8.16(s,1H),7.62(d,J＝7.9Hz,2H),7.38(d,J＝7.9Hz,2H),7.14(s,2H),4.07-3.90(m,1H),3.90-3.76(m,1H),3.58(t,J＝3.6Hz,4H),3.49(s,2H),2.96(d,J＝11.9Hz,2H),2.66-2.53(m,2H),2.42-2.31(m,4H),1.77-1.73(m,2H),1.47-1.35(m,2H).
Example 84
Compound 61: 2-amino-4-methyl-5- (4- (morpholinomethyl) phenyl) -N- (tetrahydro-2H-pyran-4-yl) nicotinamide
Referring to the procedure of example 23, step (1) was replaced with (6-amino-5- (methoxycarbonyl) -4-methylpyridin-3-yl) boronic acid instead of (6-amino-5- (methoxycarbonyl) pyridin-3-yl) boronic acid, and the title compound (12.0 mg, white solid) was obtained through a similar procedure. MS (ESI) m/z 411.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.47(d,J＝7.9Hz,1H),7.79(s,1H),7.36(d,J＝7.9Hz,2H),7.24(d,J＝7.9Hz,2H),5.62(s,2H),4.07-3.96(m,1H),3.90-3.83(m,2H),3.59(t,J＝4.4Hz,4H),3.50(s,2H),3.41-3.32(m,J＝1.5Hz,2H),2.39-2.37(m,4H),2.08(s,3H),1.87-1.79(m,2H),1.54-1.42(m,2H).
Example 85
Compound 67:6- (4- (1-ethylpiperidin-4-yl) phenyl) -N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -1H-indazole-3-carboxamide
Referring to the procedure of example 18, step (3) was performed as 1-ethyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenyl) piperidine and 6-bromo-N- (4-hydroxy bicyclo [ 2.2.2.2)]Oct-1-yl) -1H-indazole-3-carboxamide (prepared from 6-bromo-1H-indazole-3-carboxylic acid and 4-aminobicyclo [2.2.2 ]Octane-1-ol hydrochloride condensation) to give the title compound (2.3 mg, white solid). MS (ESI) m/z 473.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ13.60(brs,1H),8.17(d,J＝8.6Hz,1H),7.75(s,1H),7.67(d,J＝7.7Hz,2H),7.53(d,J＝8.9Hz,1H),7.37(d,J＝7.9Hz,2H),7.28(s,1H),4.36(s,1H),3.71-3.54(m,1H),3.04-3.00(m,2H),2.42-2.37(m,2H),2.13-2.06(m,6H),2.04-1.98(m,2H),1.83-1.76(m,2H),1.75-1.69(m,2H),1.68-1.64(m,6H),1.05(t,J＝7.0Hz,3H).
Example 86
Compound 68: (R) -2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinamide
(1) (R) -3- (5-bromo-1H-indazol-1-yl) pyrrolidine-1-carboxylic acid tert-butyl ester
5-bromo-1H-indazole (1.00 g,5.08 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), cooled to 0 ℃, tert-butyl (S) -3-hydroxypyrrolidine-1-carboxylate (1.14 g,6.09 mmol) and triphenylphosphine (1.59 g,6.09 mmol) were added, diethyl azodicarboxylate (1.06 g,6.09 mmol) was added under nitrogen, and the mixture heated to 50℃and stirred for 16 hours. The reaction solution was cooled to room temperature, quenched with water, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate=5/1) to give the title compound (500 mg, white solid), yield: 26.9%. MS (ESI) m/z 366.2[ M+H ]] + .
(2) (R) -5-bromo-1- (pyrrolidin-3-yl) -1H-indazole
The procedure of example 43, step (2), was followed using (R) -3- (5-bromo-1H-indazol-1-yl) pyrrolidine-1-carboxylic acid tert-butyl ester as starting material to give the title compound (320 mg, white solid). MS (ESI) m/z 266.1[ M+H ] ] + .
(3) (R) -5-bromo-1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazole
The procedure of step (3) was followed, using (R) -5-bromo-1- (pyrrolidin-3-yl) -1H-indazole as a starting material, to give the title compound (210 mg, white solid). MS (ESI) m/z 350.1[ M+H ]] + .
(4) (R) -2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinic acid methyl ester
The procedure of example 43, step (4), was followed using (R) -5-bromo-1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazole and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate as starting materials to give the title compound (160 mg, white solid). MS (ESI) m/z 422.2[ M+H ]] + .
(5) (R) -2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinic acid
The procedure of example 43, step (5), was followed using (R) -2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinic acid methyl ester as starting material to give the title compound (100 mg, white solid). MS (ESI) m/z 408.2[ M+H ]] + .
(6) (R) -2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinamide
Reference to example 43 procedure (6) nicotinic acid as (R) -2-amino-5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) and 4-amino-bicyclo [ 2.2.2.2 ]Starting from oct-1-ol hydrochloride, the title compound (36.4 mg, white solid) was obtained. MS (ESI): M/z531.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.40(d,J＝2.3Hz,1H),8.10(s,1H),8.05(d,J＝2.3Hz,1H),7.98(s,1H),7.91(d,J＝8.9Hz,1H),7.79(s,1H),7.71(dd,J＝8.9,1.5Hz,1H),6.89(s,2H),5.44-5.33(m,1H),4.30(s,1H),3.91-3.81(m,2H),3.35-3.30(m,2H),3.16(t,J＝8.7Hz,1H),2.91-2.82(m,2H),2.80-2.74(m,1H),2.39-2.30(m,2H),2.26-2.16(m,1H),2.12-2.03(m,6H),1.80(d,J＝11.4Hz,2H),1.68-1.59(m,6H),1.52-1.36(m,2H).
Example 87
Compound 69: (S) -2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (1- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -1H-indazol-5-yl) nicotinamide
Reference to the procedure of example 86, 5-bromo-1H-indoleThe title compound (126.2 mg, white solid) was obtained in a similar procedure starting from oxazole and tert-butyl (R) -3-hydroxypyrrolidine-1-carboxylate. MS (ESI) m/z531.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.40(d,J＝2.4Hz,1H),8.10(s,1H),8.05(d,J＝2.4Hz,1H),7.99(d,J＝0.7Hz,1H),7.91(d,J＝8.8Hz,1H),7.80(s,1H),7.71(dd,J＝8.8,1.6Hz,1H),6.88(s,2H),5.46-5.33(m,1H),4.31(s,1H),3.89-3.82(m,2H),3.35-3.30(m,2H),3.16(t,J＝8.8Hz,1H),2.91-2.82(m,2H),2.80-2.74(m,1H),2.45-2.30(m,2H),2.27-2.18(m,1H),2.08-2.04(m,6H),1.81(d,J＝11.7Hz,2H),1.65-1.61(m,6H),1.50-1.38(m,2H).
Example 88
Compound 70: (R) -2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinamide
(1) (R) -3- (5-bromo-2H-indazol-2-yl) pyrrolidine-1-carboxylic acid tert-butyl ester
5-bromo-2-nitrobenzaldehyde (10.0 g,43.48 mmol) was dissolved in isopropanol (100 mL), tert-butyl (R) -3-aminopyrrolidine-1-carboxylate (8.90 g,47.83 mmol) and tributylphosphine (26.3 g,130.44 mmol) were added and the mixture was heated to 80℃under nitrogen atmosphere and stirred for 15 hours. The reaction solution was cooled to room temperature, quenched with water, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate=5/1) to give the title compound (9.7 g, white solid), yield: 61.0%. MS (ESI) m/z 366.2[ M+H ] ] + .
(2) (R) -5-bromo-2- (pyrrolidin-3-yl) -2H-indazole
The procedure of step (1) was followed using (R) -3- (5-bromo-2H-indazol-2-yl) pyrrolidine-1-carboxylic acid tert-butyl ester as starting material in accordance with example 44 to give the title compound (7.1 g, white solid). MS (ESI) m/z 266.2[ M+H ]] + .
(3) (R) -5-bromo-2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazole
The procedure of step (2) was followed using (R) -5-bromo-2- (pyrrolidin-3-yl) -2H-indazole as a starting material in accordance with example 44 to give the title compound (2.1 g, white solid). MS (ESI) m/z 350.2[ M+H ]] + .
(4) (R) -2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid methyl ester
The procedure of step (3) of example 44 was followed using (R) -5-bromo-2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazole and methyl 2-amino-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate as starting materials to give the title compound (280 mg, white solid). MS (ESI) m/z 422.2[ M+H ]] + .
(5) (R) -2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid
The procedure of example 44, step (4), was followed using (R) -2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid methyl ester as starting material to give the title compound (200 mg, white solid). MS (ESI) m/z 408.2[ M+H ] ] + .
(6) (R) -2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinamide
Reference example 44 procedure of step (5) was followed using (R) -2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid and 4-aminobicyclo [ 2.2.2.2]Starting from oct-1-ol hydrochloride, the title compound (198.9 mg, white solid) was obtained. MS (ESI): M/z531.4[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.47(s,1H),8.39(d,J＝2.4Hz,1H),8.03(d,J＝2.4Hz,1H),7.92(s,1H),7.82(s,1H),7.70-7.66(m,1H),7.57(dd,J＝9.0,1.7Hz,1H),6.87(s,2H),5.27-5.17(m,1H),4.31(s,1H),3.92-3.81(m,2H),3.35-3.30(m,2H),3.15-3.06(m,1H),3.02-2.89(m,2H),2.71-2.65(m,1H),2.48-2.41(m,1H),2.37-2.30(m,1H),2.28-2.18(m,1H),2.09-2.03(m,6H),1.82(d,J＝11.8Hz,2H),1.66-1.60(m,6H),1.50-1.36(m,2H).
Example 89
Compound 71: (S) -2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinamide
In a similar procedure as defined in example 88, starting from 5-bromo-2-nitrobenzaldehyde and tert-butyl (S) -3-aminopyrrolidine-1-carboxylate, the title compound (195.2 mg, white solid) was obtained. MS (ESI) m/z 531.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.47(s,1H),8.39(d,J＝2.2Hz,1H),8.03(d,J＝2.1Hz,1H),7.92(s,1H),7.82(s,1H),7.70-7.66(m,1H),7.58(dd,J＝9.0,1.4Hz,1H),6.88(s,2H),5.27-5.17(m,1H),4.31(s,1H),3.90-3.83(m,2H),3.35-3.30(m,2H),3.15-3.04(m,1H),3.03-2.92(m,2H),2.73-2.65(m,1H),2.48-2.41(m,1H),2.39-2.31(m,1H),2.28-2.20(m,1H),2.09-2.03(m,6H),1.81(d,J＝11.8Hz,2H),1.66-1.60(m,6H),1.48-1.39(m,2H).
Example 90
Compound 72: 2-amino-N- (trans-4-hydroxycyclohexyl) -5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinamide
Referring to the procedure of example 44, step (5) was performed using 2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid and trans-4-aminocyclohexane-1-ol as starting materials, and yielded the title compound (1.5 mg, white solid) through similar procedures. MS (ESI) M/z505.3[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.48(s,1H),8.44-8.41(m,1H),8.34(d,J＝7.8Hz,1H),8.17(d,J＝2.2Hz,1H),7.93(s,1H),7.69(d,J＝9.0Hz,1H),7.59(d,J＝9.0Hz,1H),7.09(s,2H),5.27-5.15(m,1H),4.59(d,J＝3.7Hz,1H),3.89-3.83(m,2H),3.80-3.68(m,1H),3.42-3.40(m,1H),3.31-3.29(m,1H),3.13-3.07(m,1H),3.02-2.89(m,3H),2.70-2.68(m,1H),2.49-2.41(m,1H),2.39-2.30(m,1H),2.28-2.17(m,1H),1.91-1.78(m,6H),1.48-1.35(m,4H),1.30-1.20(m,2H).
Example 91
Compound 74: 2-amino-N- (tetrahydro-2H-pyran-4-yl) -5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinamide
Referring to the procedure of example 44, step (5) was performed using 2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid and tetrahydro-2H-pyran-4-amine as starting materials, and yielded the title compound (1.8 mg, white solid) via similar procedure. MS (ESI) m/z 491.3[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.50-8.43(m,3H),8.21(d,J＝1.8Hz,1H),7.94(s,1H),7.72-7.68(d,J＝9.2Hz,1H),7.62-7.57(d,J＝9.2Hz,1H),7.10(s,2H),5.27-5.18(m,1H),4.07-3.98(m,1H),3.93-3.83(m,4H),3.43-3.39(m,2H),3.31-3.29(m,1H),3.14-3.07(m,1H),3.02-2.86(m,3H),2.71-2.68(m,1H),2.48-2.41(m,1H),2.37-2.30(m,1H),2.28-2.19(m,1H),1.82-1.78(m,4H),1.64-1.55(m,2H),1.50-1.36(m,2H).
Example 92
Compound 76: 2-amino-N- ((3 r,6 s) -6- (hydroxymethyl) tetrahydro-2H-pyran-3-yl) -5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinamide
Referring to the procedure of example 44, step (5) was performed using 2-amino-5- (2- (1- (tetrahydro-2H-pyran-4-yl) pyrrolidin-3-yl) -2H-indazol-5-yl) nicotinic acid and ((2 s,5 r) -5-aminotetrahydro-2H-pyran-2-yl) methanolic hydrochloride as starting materials, and the title compound (20.5 mg, white solid) was obtained by similar procedures. MS (ESI) m/z 521.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.48(s,1H),8.44(d,J＝2.3Hz,1H),8.33(d,J＝7.3Hz,1H),8.19(d,J＝2.2Hz,1H),7.93(s,1H),7.70(d,J＝9.0Hz,1H),7.63-7.56(m,1H),7.11(s,2H),5.27-5.16(m,1H),4.68-4.63(m,1H),3.97-3.81(m,5H),3.43-3.38(m,2H),3.27-3.17(m,2H),3.16-3.08(m,2H),3.02-2.92(m,2H),2.28-2.22(m,1H),2.03-1.97(m,1H),1.86-1.70(m,4H),1.68-1.39(m,4H),1.36-1.21(m,2H).
Example 93
Compound 100: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (1- (tetrahydro-2H-pyran-4-yl) piperidin-4-yl) -2H-indazol-5-yl) nicotinamide
In a similar manner to that of example 46, step (2) was repeated using 5-bromo-2- (piperidin-4-yl) -2H-indazole hydrochloride and tetrahydro-4H-pyran-4-one as a starting material to give the title compound (5.9 mg, white solid). MS (ESI) m/z 545.4[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.45(s,1H),8.38(d,J＝2.2Hz,1H),8.03(d,J＝2.1Hz,1H),7.90(s,1H),7.82(s,1H),7.67(d,J＝8.9Hz,1H),7.57(dd,J＝9.0,1.5Hz,1H),6.87(s,2H),4.53-4.41(m,1H),4.31(s,1H),3.91(dd,J＝10.5,3.3Hz,2H),3.31-3.26(m,2H),3.05(d,J＝11.5Hz,2H),2.69-2.67(m,1H),2.36-2.30(m,2H),2.17-2.08(m,4H),2.08-2.02(m,6H),1.72(d,J＝12.3Hz,2H),1.67-1.59(m,6H),1.55-1.40(m,2H).
Example 94
Compound 146: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinolin-7-yl) nicotinamide
(1) 7-bromo-2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinoline
7-bromo-1, 2,3, 4-tetrahydroisoquinoline hydrochloride (1.0 g,4.04 mmol), 4-bromotetrahydro-2H-pyran (800 mg,4.85 mmol) and potassium carbonate (2.79 g,20.20 mmol) are dissolved in acetonitrile (20 mL) and the mixture is heated to 80℃under nitrogen and stirred for 16 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=20/1) to give the title compound (127 mg, yellow oil), yield: 10.1%. MS (ESI) m/z 296.0[ M+H ]] + .
(2) 2- (tetrahydro-2H-pyran-4-yl) -7- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1,2,3, 4-tetrahydroisoquinoline
7-bromo-2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinoline (80 mg,0.27 mmol) was dissolved in 1, 4-dioxane (10 mL) and pinacol bisborate (76 mg,0.30 mmol) was added, [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride (100 mg,0.14 mmol) and potassium acetate (53 mg,0.54 mmol). The mixture was heated to 90 ℃ under nitrogen and stirred for 3 hours. The reaction solution was used directly in the next step. MS (ESI) m/z 344.2[ M+H ] ] + .
(3) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (tetrahydro-2H-pyran-4-yl) -1,2,3, 4-tetrahydroisoquinolin-7-yl) nicotinamide
Intermediate 5 (149 mg,0.44 mmol) was added to the reaction solution of step (2), followed by addition of [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (295 mg,0.20 mmol), potassium carbonate (165 mg,1.20 mmol) and pure water (1 mL). The mixture was heated to 100 ℃ under nitrogen and stirred overnight. The reaction was purified by Pre-HPLC to give the title compound (14.8 mg, white solid), yield: 7.8%. MS (ESI) M/z477.1[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.32(d,J＝2.4Hz,1H),7.96(d,J＝2.0Hz,1H),7.76(s,1H),7.40(d,J＝8.0Hz,1H),7.33(s,1H),7.14(d,J＝8.0Hz,1H),6.87(s,2H),4.29(s,1H),3.93(dd,J＝11.2,8.0Hz,2H),3.75(s,2H),3.39-3.36(m,2H),2.80-2.77(m,4H),2.64-2.57(m,1H),2.07-2.03(m,6H),1.81-1.78(m,2H),1.64-1.60(m,6H),1.57-1.48(m,2H).
Example 95
Compound 150: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (1-isopropylpiperidin-4-yl) -2H-indazol-5-yl) nicotinamide
(1) 5-bromo-2- (1-isopropylpiperidin-4-yl) -2H-indazole
5-bromo-2- (piperidin-4-yl) -2H indazole hydrochloride (220 mg,0.79mmol, example 46, step 1), 2-bromopropane [ (A)116mg,0.94 mmol) and potassium carbonate (543 mg,3.93 mmol) were dissolved in acetonitrile (20 mL) and the mixture was heated to 80℃under nitrogen and stirred for 12 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane/methanol=10/1) to give the title compound (80 mg, white solid), yield: 31.6%. MS (ESI) m/z 322.1[ M+H ] ] + .
(2) 2- (1-isopropylpiperidin-4-yl) -5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -2H-indazole
5-bromo-2- (1-isopropylpiperidin-4-yl) -2H-indazole (80 mg,0.25 mmol) was dissolved in 1, 4-dioxane (10 mL), and pinacol biborate (69 mg,0.27 mmol) and [1,1' -bis (diphenylphosphine) ferrocene were added]Palladium dichloride (92 mg,0.12 mmol) and potassium acetate (49 mg,0.50 mmol). The mixture was heated to 90 ℃ under nitrogen and stirred for 3 hours. The reaction solution was used directly in the next step. MS (ESI) m/z 370.2[ M+H ]] + .
(3) 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (1-isopropylpiperidin-4-yl) -2H-indazol-5-yl) nicotinamide
Intermediate 5 (91 mg,0.27 mmol) was added to the reaction mixture of step (2) followed by [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (90 mg,1.22 mmol), potassium carbonate (68 mg,0.49 mmol) and pure water (1 mL), and the mixture was heated to 100deg.C under nitrogen and stirred overnight. The reaction was purified by Pre-HPLC to give the title compound (4.0 mg, white solid), yield: 3.3%. MS (ESI) m/z 503.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.44(s,1H),8.37(d,J＝2.4Hz,1H),8.02(d,J＝2.4Hz,1H),7.90(s,1H),7.81(s,1H),7.68-7.66(m,1H),7.58-7.56(m,1H),6.86(s,2H),4.46(s,1H),4.29(s,1H),2.98-2.94(m,2H),2.78-2.75(m,1H),2.12-1.94(m,12H),1.64-1.60(m,6H),1.06-1.03(m,6H).
Example 96
Compound 151: 2-amino-N- (4-hydroxy-bicyclo [2.2.2] oct-1-yl) -5- (2- (1-methylpiperidin-4-yl) -2H-indazol-5-yl) nicotinamide
The title compound (5.0 mg, white solid) was obtained by similar procedure using the method of example 95 starting from 5-bromo-2- (piperidin-4-yl) -2H indazole hydrochloride (step 1 of example 46) and iodomethane. MS (ESI) m/z 475.1[ M+H ] ] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.47(s,1H),8.41(d,J＝2.8Hz,1H),8.05(d,J＝3.2Hz,1H),7.92(s,1H),7.84(s,1H),7.71(d,J＝12.0Hz,1H),7.61-7.57(m,1H),6.89(s,2H),4.50-4.45(m,1H),4.33(s,1H),2.94-2.89(m,2H),2.26(s,3H),2.15-2.05(m,12H),1.67-1.62(m,6H).
Example 97
Compound 147: 2-amino-N- (4-hydroxy bicyclo [2.2.2] oct-1-yl) -5- (1 '- (oxa-3-yl) -2, 3-dihydro-spiro [ inden-1, 3' -pyrrolidin ] -5-yl) nicotinamide
Referring to the procedure of example 67, steps (5) to (8), step (5) replaces tetrahydro-4H-pyran-4-one with 3-oxetanone and step (8) uses 4-aminobicyclo [2.2.2]In place of tetrahydro-2H-pyran-4-amine, octan-1-ol was used in a similar procedure to give the title compound (2.2 mg, white solid). MS (ESI) m/z 489.2[ M+H ]] + . 1 H NMR(400MHz,DMSO-d 6 )δ8.30(d,J＝2.0Hz,1H),7.96(d,J＝2.4Hz,1H),7.74(s,1H),7.44-7.42(m,2H),7.33(d,J＝7.6Hz,1H),6.87(s,2H),4.59-4.56(m,2H),4.51-4.47(m,2H),4.29(s,1H),3.66-3.63(m,1H),2.90-2.86(m,2H),2.72-2.63(m,3H),2.16-2.13(m,1H),2.05-1.94(m,10H),1.63-1.57(m,6H).
Test example 1: kinase Activity assay
ALK2 ADP-Glo test method
The final initial concentration of compound was 10um, a total of 10 concentrations at 3-fold gradient dilution. The compound was 50-fold diluted in 1X kinase reaction buffer and shaken on a shaker for 20 minutes. Preparation of 2X kinase with 1X enzyme reaction buffer. mu.L of kinase (Carna, cat. No. 09-134) was added to each well of the reaction plate. 1. Mu.L of the compound diluted in buffer was added to each well, and the plates were sealed with a sealing plate membrane and centrifuged at 1000g for 30 seconds and incubated at 25℃for 10 secondsAnd (3) minutes. Preparation of 4X ATP with 1X enzyme reaction buffer&Casein (Signalchem Co., ltd., product No. C03-54BN-1 MG) was mixed with 1. Mu.L of 4 XATP was added to the reaction plate&Casein mixed solution. Plates were sealed with a sealing plate membrane and centrifuged at 1000g for 30 seconds and incubated at 25℃for 60 minutes. Transfer 4. Mu.L ADP-Glo (kit from Promega, cat. V9102) to 384 reaction plates at 1000rpm/min, centrifuge 1min, incubate at 25℃for 40min. Transfer 8. Mu.L of Detection solution to 384 reaction plates at 1000rpm/min, centrifuge 1min, incubate at 25℃for 40min. RLU signals were read using a BGM microplate reader. Data analysis and calculation of IC using graphpad7.0 software 50 。
ALK5 ADP-Glo test method
The final initial concentration of compound was 10um, a total of 10 concentrations at 3-fold gradient dilution. The compound was 50-fold diluted in 1X kinase reaction buffer and shaken on a shaker for 20 minutes. Preparation of 2X kinase with 1X enzyme reaction buffer. mu.L of kinase (ThermoFisher company, cat. No. 1795372T) was added to each well of the reaction plate. To each well 1. Mu.L of the diluted compound in buffer was added, and the plates were blocked with a plate membrane and centrifuged at 1000g for 30 seconds and incubated at 25℃for 10 minutes. A4 XATP mixture was prepared with 1 Xenzyme reaction buffer, and 1. Mu.L of the 4 XATP mixture was added to the reaction plate. Plates were sealed with a sealing plate membrane and centrifuged at 1000g for 30 seconds and incubated at 25℃for 180 minutes. Transfer 4. Mu.L ADP-Glo (kit from Promega, cat. V9101) to 384 reaction plates at 1000rpm/min, centrifuge 1min, incubate at 25℃for 40min. Transfer 8. Mu.L of Detection solution to 384 reaction plates at 1000rpm/min, centrifuge 1min, incubate at 25℃for 40min. RLU signals were read using a BGM microplate reader. Data analysis and calculation of IC using graphpad7.0 software 50 。
The IC50 values for the enzyme activity of each compound were obtained using the analytical software XLfit5.5.5 fit-up response curve.
Wherein the positive compound INCB-00928 was prepared according to example 34 of patent WO 2018014829.
The results are shown in Table 1, wherein A represents < 5nM, B represents 5-20nM, C represents 20-100nM, D represents 100-500nM, E represents 500-1000nM, and F represents > 1000nM.
TABLE 1
Test example 2: cell testing method
The compounds were tested for their inhibitory activity on the mRNA expression level of Hepcidin genes in HepG2 cells using qPCR. Will be 3.5X10 5 Individual HepG2 cells were seeded in 12-well plates. The culture was carried out for 24 hours by adherence. Compounds of different concentrations were added to 12-well plates at a maximum final concentration of 10 μm, 2-fold dilution, 10 concentration points. The compound was incubated for 2 hours, then BMP6 (from R&Company D, cat# 507-BP-020), final concentration of BMP6 was 10ng/mL. Cells were incubated for 6 hours in the presence of compound and BMP 6. After removing the medium from the 12-well plate and washing the well with pre-chilled PBS, cells were digested with trypsin and centrifuged at 1200rpm at 4℃for 5 minutes to collect a cell sample. The obtained cell sample is used for extracting RNA, and q-PCR method is used for testing, and detecting gene Hepcidin (HAMP)](primer was obtained from General Biosystems company) and GUSB was used as the reference gene. Analyzing the data result by using Graph Pad software and calculating IC 50 . The results of each compound are shown in table 2, wherein+++ represents in the range of < 100nM, ++ means 100-1000nM, + means > 1000nM.
TABLE 2
/>

Claims (52)

1. A compound having a structure represented by general formula (II), a deuterated, a stereoisomer, or a pharmaceutically acceptable salt thereof:
X 1 、X 2 CH;
R 1 selected from the group consisting ofOptionally by 1-3R 5 Substitution;
R 5 selected from hydroxy, - (CR) a R b b) n -OH；
Ring B is selected from
R 2 Is a 5-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O and optionally containing 1-3R 6 Substitution;
R 6 selected from C 1-6 Alkyl, 4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl containing 1O atom, optionally substituted with 1-3R e Substitution;
R a 、R b independently selected from H, C 1-3 An alkyl group;
R e selected from halogen;
n is an integer from 1 to 3.
2. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 1, wherein:
R 1 selected from the group consisting of
R 5 Selected from hydroxy, -CH 2 OH、-C(CH 3 ) 2 OH。
3. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 2 having a structure according to formula (II-1):
ring B is selected from
R 2 Is a 5-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O and optionally containing 1-3R 6 Substitution;
R 6 selected from C 1-6 Alkyl, 4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl containing 1O atom, optionally substituted with 1-3R e Substitution;
R e selected from halogen.
4. A compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 3, wherein:
R 2 Selected from the group consisting ofAnd optionally is substituted with 1R 6 Substitution;
R 6 selected from C 1-3 Alkyl, 4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl containing 1O atom, optionally substituted with 1-3R e Substitution;
R e selected from halogen.
5. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 4, wherein:
R 2 selected from the group consisting of
6. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 5, wherein:
R 2 selected from the group consisting ofAnd is covered with 1R 6 Substitution;
R 6 is 4-6 membered heterocycloalkyl containing 1An O atom.
7. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 6, wherein:
R 2 selected from the group consisting of
8. The compound of claim 7, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof, wherein:
R 6 selected from the group consisting of
9. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 8, wherein:
R 6 selected from the group consisting of
10. A compound having a structure represented by general formula (III), a deuterated compound, a stereoisomer, or a pharmaceutically acceptable salt thereof, characterized by having a structure represented by formula (III):
Ring B is selected from
R 1 Selected from the group consisting ofR 1 Optionally by 1-3R 5 Substitution;
R 5 selected from hydroxy, -CH 2 OH、-C(CH 3 ) 2 OH；
L 2 Selected from chemical bonds, - (CR) a R b ) n -；
R 2 Heterocyclylalkyl containing 1-2 heteroatoms selected from N, O for 5-6 members, optionally substituted with 1-2R 6 Substitution;
R 6 selected from halogen, or
R 6 Selected from 4-7 membered cycloalkyl, 4-7 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, C 1-3 Alkyl, and optionally substituted with 1-3R e Substitution;
R a 、R b independently selected from H or C 1-3 An alkyl group;
R e is halogen;
n is an integer of 1 to 3.
11. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 10, wherein:
R 1 selected from the group consisting of
12. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 10, wherein:
ring B is selected from
L 2 Selected from chemical bonds, C 1-3 An alkylene group;
R 2 heterocyclylalkyl containing 1-2 heteroatoms selected from N, O for 5-6 members, optionally substituted with 1-2R 6 Substitution; r is R 6 Selected from halogen, or
R 6 Selected from the group consisting of4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O, C 1-3 Alkyl, and optionally substituted with 1-3R e And (3) substitution.
13. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 10, wherein:
Ring B is selected from
R 1 Selected from the group consisting ofOptionally by 1R 5 Substitution;
R 5 selected from hydroxy, -CH 2 OH；
L 2 Selected from the group consisting of chemical bonds;
R 2 heterocyclylalkyl containing 1-2 heteroatoms selected from N, O for 5-6 members, optionally substituted with 1R 6 Substitution;
R 6 selected from 4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl containing 1O, C 1-3 Alkyl, and optionally substituted with 1-3R e Substitution; r is R e Is halogen.
14. The compound of claim 13, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof, wherein:
R 2 selected from the group consisting of
R 6 Selected from the group consisting ofCyclohexyl, C 1-3 An alkyl group.
15. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 14, wherein:
R 2 selected from the group consisting of
16. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 14, wherein:
R 6 selected from the group consisting ofC 1-3 An alkyl group.
17. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 13 or 14, wherein:
R 1 and R is R 5 The following structure is formed:
18. the compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 10, wherein:
ring B is selected from
R 1 Selected from the group consisting ofOptionally by 1R 5 Substitution;
R 5 selected from hydroxy, -CH 2 OH；
L 2 Selected from the group consisting of chemical bonds;
R 2 heterocyclylalkyl containing 1-2N for 5-6 membered ring, substituted with 1R 6 Substitution;
R 6 selected from 4-6 membered heterocycloalkyl containing 1-2 heteroatoms selected from N, O.
19. The compound of claim 18, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof, wherein:
R 2 selected from the group consisting of
R 6 Selected from the group consisting of
20. The compound of claim 19, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof, wherein:
R 2 selected from the group consisting of
21. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 19 wherein
R 6 Selected from the group consisting of
22. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 18 or 19, wherein:
R 1 and R is R 5 The following structure is formed:
23. the compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 10, wherein:
ring B is selected from
R 1 Selected from the group consisting ofOptionally by 1R 5 Substitution;
R 5 selected from hydroxy, -CH 2 OH；
L 2 Selected from chemical bonds, C 1-3 An alkylene group;
R 2 heterocyclylalkyl containing 1-2 heteroatoms selected from N, O for 5-6 members, optionally substituted with 1-2R 6 Substitution;
R 6 Selected from halogen, or
R 6 Selected from 4-6 membered cycloalkyl, 4-6 membered heterocycloalkyl containing 1O, and optionally substituted with 1-3R e Substitution;
R e is halogen.
24. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 23, wherein:
R 2 selected from the group consisting ofR 6 Selected from halogen, or
R 6 Selected from the group consisting ofCyclohexyl, and optionally substituted with 1-3R e Substitution; r is R e Is halogen.
25. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 24, wherein:
R 2 selected from the group consisting of
26. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 24, wherein:
R 6 selected from the group consisting of
27. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 23 or 24, wherein:
R 1 and R is R 5 The following structure is formed:
28. a compound having a structure represented by general formula (IV), a deuterated, a stereoisomer, or a pharmaceutically acceptable salt thereof:
X 1 、X 2 CH;
R 1 selected from the group consisting ofOptionally by 1-2R 5 Substitution;
R 5 selected from-C 1-3 alkyl-OH, OH;
ring B isq is 1R is an integer of 1 to 3; z is Z 1 、Z 2 Or Z is 3 CH; />Is a single bond or a double bond; when->Z is a single bond 4 Is CH 2 Or O; when->In the case of double bonds, Z 4 CH;
L 2 is a chemical bond, - (CR) a R b ) n -；
R 2 Selected from H, or
R 2 Selected from 4-6 membered heterocycloalkyl containing 1-2O atoms;
R a 、R b independently selected from H or C 1-3 An alkyl group;
n is an integer of 1 to 3.
29. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 28, wherein:
R 1 selected from the group consisting of
30. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 28, wherein:
R 2 selected from the group consisting of
31. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 30, wherein:
R 2 selected from the group consisting of
32. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 28, wherein:
ring B is selected from
33. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 28, wherein:
ring B isr is an integer from 1 to 2, < >>Is a single bond or a double bond.
34. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 33, wherein:
ring B is* Terminal to pyridine, terminal to L 2 And r is an integer of 1 to 2,is a single bond or a double bond;
L 2 is a chemical bondOr C 1-3 An alkylene group;
R 2 selected from H, or
R 2 Selected from 4-6 membered heterocycloalkyl containing 1-2O atoms.
35. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 34, wherein:
ring B is
36. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 34, wherein:
R 2 is that
37. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 36, wherein:
R 2 is that
38. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 33, wherein:
R 1 selected from the group consisting ofOptionally by 1-2R 5 Substitution;
R 5 selected from-OH, -C 1-3 alkyl-OH.
39. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 38, wherein:
R 1 selected from the group consisting of
40. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 38, wherein:
R 5 selected from-OH, -CH 2 -OH。
41. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 38, wherein:
R 1 And R is R 5 The following structure is formed:
42. the compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 28, wherein:
ring B is selected from * Terminal to pyridine, terminal to L 2 Connecting;
L 2 is a chemical bond, C 1-3 An alkylene group;
R 2 is H or
43. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 42 wherein:
ring B is selected from
44. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 42 wherein:
L 2 is a chemical bond or isopropyl.
45. The compound, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 44 wherein:
R 2 is H or
46. A compound of the structure, deuterated, stereoisomer, or pharmaceutically acceptable salt thereof:
/>
/>
47. a pharmaceutical composition comprising a compound of any one of claims 1-46, a deuterated, a stereoisomer, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier therefor.
48. Use of a compound according to any one of claims 1 to 46, a deuterated, a stereoisomer, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 47, for the manufacture of a medicament for the treatment and/or prophylaxis of an ALK 2-related disorder.
49. The use according to claim 48, wherein the ALK 2-related diseases are anaemia, inflammation, neoplasm and a portion of ALK 2-related genetic diseases.
50. The use according to claim 49, wherein the ALK 2-associated condition is progressive fibrodysplasia, diffuse-in type bridge glioma, iron refractory iron deficiency anemia, inflammatory anemia, myelodysplastic syndrome, multiple myeloma, myeloproliferative tumor-associated anemia.
51. The use according to claim 48 wherein the medicament is used alone or in combination with other therapeutic agents.
52. The use according to claim 51, wherein the additional therapeutic agent is a JAK2 inhibitor for the treatment of myeloproliferative neoplasm-related anemia.
CN202310387713.7A 2022-04-13 2023-04-12 ALK2 kinase inhibitors Active CN116478145B (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CN202210388938 2022-04-13
CN202210388938X 2022-04-13
CN2022109364563 2022-08-05
CN202210936456 2022-08-05

Publications (2)

Publication Number Publication Date
CN116478145A CN116478145A (en) 2023-07-25
CN116478145B true CN116478145B (en) 2024-02-02

Family

ID=87217109

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202310387713.7A Active CN116478145B (en) 2022-04-13 2023-04-12 ALK2 kinase inhibitors

Country Status (2)

Country Link
CN (1) CN116478145B (en)
WO (1) WO2023198114A1 (en)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109641871A (en) * 2016-07-20 2019-04-16 诺华股份有限公司 Aminopyrazole derivatives and its alternatively purposes of property ALK-2 inhibitor
WO2019227059A1 (en) * 2018-05-25 2019-11-28 Silverback Therapeutics, Inc. Amino-pyrazinecarboxamide compounds, conjugates, and uses thereof
WO2021102258A1 (en) * 2019-11-22 2021-05-27 Incyte Corporation Combination therapy comprising an alk2 inhibitor and a jak2 inhibitor
WO2021248081A1 (en) * 2020-06-05 2021-12-09 The J. David Gladstone Institutes, A Testamentary Trust Established Under The Will Of J. David Gladstone Acvr1 (alk2) receptor inhibition to treat neurological diseases
WO2021257532A1 (en) * 2020-06-16 2021-12-23 Incyte Corporation Alk2 inhibitors for the treatment of anemia

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018183633A1 (en) * 2017-03-29 2018-10-04 University Of Houston System Compositions for use in methods of inhibiting protein kinases
US11026947B2 (en) * 2017-04-27 2021-06-08 The Brigham And Women's Hospital, Inc. ALK2 inhibitors and methods for inhibiting BMP signaling
AR112027A1 (en) * 2017-06-15 2019-09-11 Biocryst Pharm Inc ALK 2 KINASE INHIBITORS CONTAINING IMIDAZOLE
US11046699B2 (en) * 2018-06-05 2021-06-29 Rapt Therapeutics, Inc. Pyrazolo-pyrimidin-amino-cycloalkyl compounds and their therapeutic uses
WO2022184152A1 (en) * 2021-03-03 2022-09-09 劲方医药科技(上海)有限公司 Fused ring substituted six-membered heterocyclic compound, preparation method therefor and use thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109641871A (en) * 2016-07-20 2019-04-16 诺华股份有限公司 Aminopyrazole derivatives and its alternatively purposes of property ALK-2 inhibitor
WO2019227059A1 (en) * 2018-05-25 2019-11-28 Silverback Therapeutics, Inc. Amino-pyrazinecarboxamide compounds, conjugates, and uses thereof
WO2021102258A1 (en) * 2019-11-22 2021-05-27 Incyte Corporation Combination therapy comprising an alk2 inhibitor and a jak2 inhibitor
WO2021248081A1 (en) * 2020-06-05 2021-12-09 The J. David Gladstone Institutes, A Testamentary Trust Established Under The Will Of J. David Gladstone Acvr1 (alk2) receptor inhibition to treat neurological diseases
WO2021257532A1 (en) * 2020-06-16 2021-12-23 Incyte Corporation Alk2 inhibitors for the treatment of anemia

Also Published As

Publication number Publication date
CN116478145A (en) 2023-07-25
WO2023198114A1 (en) 2023-10-19

Similar Documents

Publication Publication Date Title
CN112437772B (en) Bcl-2 inhibitors
CN113454083A (en) KRAS mutein inhibitors
CN114174299A (en) Degradation of Bruton&#39;s Tyrosine Kinase (BTK) by conjugation of BTK inhibitors to E3 ligase ligands and methods of use
BRPI0707872A2 (en) azacyclyl substituted arylhydroisoquinolinones, the process for their preparation and their use as medicines
CN114929689A (en) Bcl-2 inhibitors
CN116323580A (en) Conjugated degradation (EGFR) of EGFR inhibitors to E3 ligase ligands and methods of use
JP2011518127A (en) Tetrahydronaphthyridines and their aza derivatives as histamine H3 receptor antagonists
TW202102497A (en) Compounds and their use in treating cancer
CN116589466A (en) Degradation of Bruton&#39;s Tyrosine Kinase (BTK) by conjugation of BTK inhibitors with E3 ligase ligands and methods of use thereof
CN115335376A (en) Degradation of Bruton&#39;s Tyrosine Kinase (BTK) by conjugation of BTK inhibitor with E3 ligase ligand and methods of use thereof
EP4269402A1 (en) Aromatic heterocyclic compound, pharmaceutical composition and use thereof
EP4351558A1 (en) Compounds having 5-(2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide as inhibitors of protein kinase phosphatase enzymes
KR20220163985A (en) Free Alkaline Crystalline Forms of Nitrogen-Containing Aromatic Derivatives
TW202237597A (en) Novel degraders of egfr
CN116478145B (en) ALK2 kinase inhibitors
WO2022253309A1 (en) Substituted heterocyclic compounds and application thereof
CN116888108B (en) Novel EGFR degradation agent
WO2023098656A1 (en) Compounds for the degradation of egfr kinase
KR20230153959A (en) Pim kinase inhibitors and methods of their use
KR20230124608A (en) Antagonists of the muscarinic acetylcholine receptor M4
WO2023138607A1 (en) Degradation of (egfr) by conjugation of egfr inhibitors with e3 ligase ligand and methods of use
CN116981675A (en) Degradation of Bruton&#39;s Tyrosine Kinase (BTK) by conjugation of BTK inhibitors to E3 ligase ligands and methods of use
CN117642157A (en) Compounds having ((3-nitrophenyl) sulfonyl) acetamide as BCL-2 inhibitors
CN116262753A (en) Novel PIM kinase inhibitors
CN115557946A (en) Heterocyclic lactam compound, pharmaceutical composition containing same and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant