CN117510463A - KIF18A inhibitor compound, pharmaceutical composition, preparation method and application thereof - Google Patents

KIF18A inhibitor compound, pharmaceutical composition, preparation method and application thereof Download PDF

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CN117510463A
CN117510463A CN202310972422.4A CN202310972422A CN117510463A CN 117510463 A CN117510463 A CN 117510463A CN 202310972422 A CN202310972422 A CN 202310972422A CN 117510463 A CN117510463 A CN 117510463A
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alkyl
compound
alkoxy
cyano
spiro
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陆标
杨方龙
张辰
金磊
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Changchun Genescience Pharmaceutical Co Ltd
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    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • A—HUMAN NECESSITIES
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    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members 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
    • C07D211/38—Halogen atoms or nitro radicals
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    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
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    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
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    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
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    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
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    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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    • 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

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Abstract

The invention provides a compound shown in a formula (I), a pharmaceutical composition, a preparation method and application thereof. The compounds have good KIF18A inhibition and can be used alone or in combination with microtubules to modulate KIF18A protein, for the treatment of KIF18A mediated disorders and/or diseases, such as tumour diseases, and for the preparation of medicaments for use in such disorders or diseases.

Description

KIF18A inhibitor compound, pharmaceutical composition, preparation method and application thereof
The present application claims priority from the following patent applications:
prior application filed by 8.5.2022 to the national intellectual property office under the patent application number 202210944271.7 entitled "KIF18A inhibitor compound, pharmaceutical composition, methods of preparation and use thereof";
prior application filed by 22 days 11 and 2022 to the national intellectual property office of China with patent application number 202211469122.6 and named as "KIF18A inhibitor compound, pharmaceutical composition and preparation method and application thereof";
the entirety of the prior application is incorporated by reference into this application.
Technical Field
The invention belongs to the field of medicines, and particularly relates to a KIF18A inhibitor compound, a pharmaceutical composition, a preparation method and application thereof.
Background
Cancer is one of the most serious diseases affecting human health, and mortality and morbidity often prevail in various disease fronts. While the quality of life of some patients has improved greatly with the continued development and advancement of medical technology and drug development, there are still more unmet clinical needs in the way of seeking drugs for effective treatment or cure of different cancers, and more new targets will provide new possibilities for future cancer drug development.
Unregulated proliferation of cancer cells occurs due to damage or loss of one or more genes that regulate the cell cycle. Various kinases and kinesins have been identified as playing a key role in the regulation and progression of cell cycle and mitosis in normally dividing cells and cancer cells.
Kinesin molecules are motor proteins that take intracellular microtubules as the rails, also called molecular motors, and can convert ATP energy into mechanical energy, and are closely related to cell mitosis and meiosis, growth and development of tissue organs, development of neurons, signal transduction, and the like in eukaryotic cells. Kinesin members share a relatively conserved motor domain. The kinesin family is broadly divided into three classes, depending on the location of the motor domain in the molecule: n-kinesins, i.e. amino groups (-NH) of the polypeptide chain of a protein 2 ) The terminal region has a motor domain; m-kinesin, i.e. a motor domain in the middle region; the C-kinesin, carboxyl (-COOH) terminal region has a motor domain.
Chromosomal instability is a hallmark of cancer, caused by chromosomal separation errors during mitosis. Targeting chromosomal instability is an emerging therapeutic strategy in drug development. KIF18A is a member of the Kinesin-8 Kinesin family of N types, and has been shown to play a role in maintaining bipolar spindle integrity and promoting the viability of chromosome instability cancer cells. Mitosis is an effective point of intervention, and many antimitotic drugs are used clinically to treat human cancers. The most widely used tubulin inhibitors, which stabilize microtubules as well as prevent microtubule assembly. Current antimitotic drugs have the limitation of a narrow therapeutic window and these problems need to be solved by developing new targets.
Although tubulin inhibitors are widely used as standard therapeutic methods for the treatment of a variety of human cancer types, these drugs present collateral damage to normal cells, including myelosuppression and neurotoxicity. Since KIF18A may not be necessary in normal diploid somatic cells (KIF 18A knockout mice are viable but have defects in reproduction, indicating that KIF18A is not an essential gene for normal somatic cell division), targeting KIF18A may significantly reduce its toxicity, which is beneficial in improving the therapeutic safety window of targeted tubulin drugs in the clinic.
KIF18A protein is highly expressed in a variety of tumors, including colon, breast, lung, pancreas, prostate, bladder, head and neck, cervical and ovarian cancers. KIF18A plays a key role in the development, progression and metastasis of breast cancer, and its high expression predicts a poor prognosis for patients. KIF18A is necessary for proliferation of chromosome instability cells derived from triple negative breast cancer or colorectal cancer, but KIF18A is not required in diploid cells. Knockout of the KIF18A gene may result in male mice being sterile, but female mice are unaffected. KIF18A mRNA expression is significantly associated with higher tumor grade and larger tumors in breast cancer patients, and KIF18A is an independent predictor of breast cancer lymph node metastasis with a risk factor of 3.2. Furthermore, inhibition of KIF18A expression not only affects key functions of KIF18A in cell mitosis, but also can reduce cancer cell migration by stabilizing leading-edge microtubules, ultimately leading to inactivation of PI3K-AKT signaling pathway to induce apoptosis.
Thus, the development of KIF18A protein inhibitors may be a new breakthrough in cancer drugs.
Disclosure of Invention
In order to solve the above technical problems, the present invention provides a compound represented by formula (I), its racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof:
Wherein X is 1 、X 2 、X 3 Identical or different, independently of one another, from N or CR 0 ；R 0 Selected from H, halogen, cyano, C 1-12 Alkyl, halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halo C 1-12 Alkoxy, cyano C 1-12 An alkoxy group;
a is selected from unsubstituted or optionally substituted with one, two or more R a Substituted with the following groups: c (C) 1-12 Alkyl, halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halo C 1-12 Alkoxy, cyano C 1-12 Alkoxy, C 3-12 Cycloalkyl, C 3-12 Cycloalkyl oxy, C 3-12 Cycloalkylthio, 3-14 membered heterocyclyl; each R a The same or different, independently of one another, are selected from halogen, cyano, C 1-12 Alkyl, halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halo C 1-12 Alkoxy, cyano C 1-12 An alkoxy group;
Y 1 、Y 2 、Y 3 the same or different, independently of one another, from N or CH;
m is selected from unsubstituted or optionally substituted with one, two or more R b Substituted with the following groups: c (C) 3-12 Cycloalkyl, C 3-12 Cycloalkenyl, 3-14 membered heterocyclyl; each R b The same or different, independently of one another, are selected from halogen, cyano, C 1-12 Alkyl, halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, cyano C 1-12 An alkoxy group;
e is selected from unsubstituted or optionally substituted with one, two or more R c Substituted with the following groups: -NH-S (=o) 2 -R c1 、-S(＝O) 2 -NH-R c2 、-S(＝O)(＝NH)-R c3 、-N(R c4 )(R c5 ) 3-14 membered heterocyclyl; each R c The same or different, independently of one another, from OH, halogen, cyano, C 1-12 Alkyl, C 1-12 Alkoxy, halo C 1-12 Alkyl, halogenated C 1-12 Alkoxy, cyano C 1-12 Alkyl, cyano C 1-12 Alkoxy, -N (R) c6 )(R c7 )；
R c1 、R c2 、R c3 、R c4 、R c5 、R c6 、R c7 Identical or different, independently of one another, from H, C 1-12 Alkyl, hydroxy C 1-12 Alkyl, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy, cyano C 1-12 Alkyl, cyano C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclyl, C 1-12 alkoxy-C 1-12 Alkyl, hydroxy C 1-12 alkyl-C 3-12 Cycloalkyl groups.
According to some embodiments, X 2 、X 3 And are not simultaneously N.
According to some embodiments, when X 1 And X 2 Is N or CR 0 When X is 3 Is CR (CR) 0 ，R 0 Selected from H, halogen, CN, C 1-6 An alkyl group; when X is 1 Is N or CH, X 2 When CH is, X 3 Is N or CR 0 ，R 0 Selected from H, halogen, CN, C 1-6 An alkyl group; when X is 1 Is N or CR 0 ，X 2 When N is N, X 3 Is N or CR 0 ；R 0 Selected from H, halogen, CN, C 1-6 An alkyl group.
According to some embodiments, A is selected from C 1-6 Alkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkyloxy, 3-8 membered heterocyclyl, halogenated C 3-8 Cycloalkyl groups.
According to some embodiments, A is selected from cyclopropyl, isopropyl,
According to some embodiments, when Y 1 When CH is, Y 2 Is CH, Y 3 Is N or CH; when Y is 1 When CH is, Y 2 Is N, Y 3 Is N or CH; when Y is 1 When N is N, Y 2 Is CH, Y 3 Is N or CH.
According to some embodiments, M is selected from C 3-12 Cycloalkyl, C 3-12 Cycloalkenyl, 3-12 membered heterocyclyl. Preferably, M is selected from C 3-12 Condensed ring cycloalkyl, C 3-12 Condensed ring cycloalkenyl, 3-12 membered condensed ring heterocyclyl, C 3-12 Spiro cycloalkyl, C 3-12 Spirocyclic cycloalkenyl, 3-12 membered spiroheterocyclic group, C 3-12 Bridged cycloalkyl, C 3-12 Bridged cycloalkenyl or 3-12 membered bridged heterocyclyl.
According to some embodiments, M is selected from
According to some embodiments, E is selected from-NH-S (=o) 2 -R c1 、-S(＝O) 2 -NH-R c2 、-S(＝O)(＝NH)-R c3 、-N(R c4 )(R c5 )、C 1-6 Alkyl substituted 3-6 membered heterocyclic group,
R c1 、R c2 、R c3 、R c4 、R c5 the same or different, are independently selected from H, hydroxy C 1-6 Alkyl, halogenated C 1-6 Alkyl, cyano C 1-6 Alkyl, C 1-6 alkyl-NH-C 1-6 Alkyl-, (C) 1-6 Alkyl group 2 N-C 1-6 Alkyl, C 3-6 cycloalkyl-NH-C 1-6 Alkyl-, C 1-6 alkoxy-C 1-6 Alkyl-, C 3-6 Cycloalkyl, 3-6 membered heterocyclyl; or alternatively
R c1 、R c2 、R c3 、R c4 、R c5 The same or different, are independently selected from H, hydroxy C 1-6 Alkyl, halogenated C 1-6 Alkyl, cyano C 1-6 Alkyl, C 1-6 alkyl-NH-C 1-6 Alkyl-, (C) 1-6 Alkyl group 2 N-C 1-6 Alkyl, C 3-6 cycloalkyl-NH-C 1-6 Alkyl-, C 1-6 alkoxy-C 1-6 Alkyl-, C 3-6 Cycloalkyl, 3-6 membered heterocyclyl, hydroxy-C 3-6 Cycloalkyl, hydroxy C 1-6 alkyl-C 3-6 Cycloalkyl groups.
According to some embodiments, E is selected from
According to some embodiments, the compound of formula (I) has the following structure:
therein, A, E, M, X 2 、X 3 、Y 1 、Y 2 、Y 3 With the definition described above.
According to some embodiments, the compound of formula (I) has a structure of formula (II), preferably a structure of formula (III), more preferably a structure of formula (IV):
therein, A, E, X 1 、X 2 、X 3 、R c1 Having the definition set forth above;represents a carbon-carbon single bond or a carbon-carbon double bond.
According to some embodiments, the compound of formula (I) is selected from the following structures:
the invention also provides a preparation method of the compound shown in the formula (I), which comprises the following steps:
(1) Reacting the compound a with the compound b to obtain a compound c;
(2) Reacting the compound c with E-H to obtain a compound shown in a formula (I);
therein, A, E, M, X 1 、X 2 、X 3 、Y 1 、Y 2 、Y 3 Independently of each other, have the definition described above; x is selected from halogen, such as Cl, br, I.
The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of a compound of formula (I), racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof.
According to an embodiment of the invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.
According to embodiments of the present invention, the pharmaceutical composition may further comprise one or more additional therapeutic agents.
The present invention also provides a method for treating a neoplastic disease, comprising administering to a patient a prophylactically or therapeutically effective amount of at least one of a compound of formula (I), racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof.
The invention also provides a method of treating a neoplastic disease comprising administering to a patient a prophylactically or therapeutically effective amount of the above pharmaceutical composition.
The tumor diseases include intestinal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer and ovarian cancer.
In some embodiments, the patient comprises a mammal, preferably a human.
The present invention also provides at least one of a compound of formula (I), racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, or a pharmaceutical composition thereof, for use in treating a neoplastic disease.
The invention also provides application of at least one of the compounds shown in the formula (I), racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof in preparing medicines.
According to an embodiment of the invention, the use may be in the manufacture of a medicament for the treatment of a KIF18A mediated disorder and/or disease, such as in the manufacture of a KIF18A inhibitor medicament.
According to an embodiment of the invention, the disease is for example a cancer, including intestinal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer or ovarian cancer.
Advantageous effects
The compounds of the invention have good inhibitory effect on KIF18A, and can be used for regulating KIF18A protein alone or in combination with microtubules to treat diseases and/or conditions mediated by KIF18A, such as tumor diseases, and for preparing medicines for treating the diseases or conditions.
Definition and description of terms
Unless otherwise indicated, the radical and term definitions recited in the specification and claims of this application, including as examples, exemplary definitions, preferred definitions, definitions recited in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. Such combinations and combinations of radical definitions and compound structures should be understood to be within the scope of the description and/or claims herein.
The numerical ranges recited in the specification and claims are equivalent to at least each specific integer number recited therein unless otherwise stated. For example, a numerical range of "1-12" corresponds to each integer number recited in the numerical range of "1-12," i.e., 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.
The term "C 1-12 Alkyl "is understood to mean straight-chain and branched alkyl radicals having 1 to 12 carbon atoms," C 1-8 Alkyl "means straight and branched alkyl having 1,2, 3, 4, 5, 6, 7, or 8 carbon atoms," C 1-6 Alkyl "means straight and branched alkyl groups having 1,2, 3, 4, 5 or 6 carbon atoms. The alkyl is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-ethylpropyl, 1, 2-dimethylpropyl, neopentyl, 1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 2, 3-dimethylbutyl, 1, 3-dimethylbutylOr 1, 2-dimethylbutyl, etc. or an isomer thereof.
The term "C 3-12 Cycloalkyl "is understood to mean a saturated monovalent monocyclic, bicyclic (e.g. fused, bridged, spiro) hydrocarbon ring or tricyclic hydrocarbon ring having 3 to 12 carbon atoms, preferably" C 3-10 Cycloalkyl ", more preferably" C 3-8 Cycloalkyl groups). The term "C 3-12 Cycloalkyl "is understood to mean a saturated monovalent monocyclic, bicyclic (e.g. bridged, spiro) hydrocarbon ring or tricyclic hydrocarbon ring having 3,4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. The C is 3-12 Cycloalkyl can be a monocyclic hydrocarbon group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group such as campholyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo [2.1.1 ]]Hexyl, bicyclo [2.2.1]Heptyl, bicyclo [2.2.1]Heptenyl, 6-dimethylbicyclo [3.1.1]Heptyl, 2, 6-trimethylbicyclo [3.1.1]Heptyl, bicyclo [2.2.2]Octyl, 2, 7-diazaspiro [3,5 ]]Nonylalkyl, 2, 6-diazaspiro [3,4 ]]Octyl, or tricyclic hydrocarbon groups such as adamantyl.
The term "C 3-12 Cycloalkenyl "is understood to mean a monovalent monocyclic, bicyclic (e.g. fused, bridged, spiro) hydrocarbon ring or tricycloolefin containing a carbon-carbon double bond, having 3 to 12 carbon atoms, preferably" C 3-10 Cycloalkenyl ", more preferably" C 3-8 A cycloalkenyl group). The term "C 3-12 Cycloalkenyl "is understood to mean a monovalent monocyclic, bicyclic (e.g. bridged, spiro) hydrocarbon ring or tricycloolefin containing a carbon-carbon double bond, having 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. The C is 3-12 The cycloalkenyl group may be a monocyclic hydrocarbon group such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclononenyl or cyclodecenyl, or a bicyclic hydrocarbon group such as spiro [2.5 ]]Oct-5-enyl, spiro [3.5 ]]Non-6-enyl, spiro [4.5 ]]Dec-7-enyl.
The term "3-14 membered heterocyclyl" refers to a saturated or unsaturated, non-aromatic ring or ring system, e.g., which is 4-, 5-, 6-, or 7-membered monocyclic, 7-, 8-, 9-, 10-, 11-, or 12-memberedOr a 10-, 11-, 12-, 13-or 14-membered tricyclic ring system, and contains at least one, for example 1, 2, 3, 4, 5 or more heteroatoms selected from O, S and N, wherein N and S can also optionally be oxidized to various oxidation states to form nitrogen oxides, -S (O) -or-S (O) 2 -a state of the device. Preferably, the heterocyclic group may be selected from "3-10 membered heterocyclic groups". The term "3-10 membered heterocyclyl" means a saturated or unsaturated, non-aromatic ring or ring system and contains at least one heteroatom selected from O, S and N. The heterocyclic group may be attached to the remainder of the molecule through any of the carbon atoms or a nitrogen atom, if present. The heterocyclic group may include fused or bridged rings as well as spiro rings. In particular, the heterocyclic groups may include, but are not limited to: 4-membered rings such as azetidinyl, oxetanyl; a 5-membered ring such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6 membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or a 7-membered ring such as diazepanyl. Optionally, the heterocyclyl may be benzo-fused. The heterocyclic group may be bicyclic, such as, but not limited to, a 5,5 membered ring, such as hexahydrocyclopenta [ c ] ]Pyrrol-2 (1H) -yl ring, or 5,6 membered bicyclic ring, e.g. hexahydropyrrolo [1,2-a ]]Pyrazin-2 (1H) -yl ring. The heterocyclic group may be partially unsaturated, i.e., it may contain one or more double bonds, such as, but not limited to, dihydrofuranyl, dihydropyranyl, 2, 5-dihydro-1H-pyrrolyl, 4H- [1,3,4]Thiadiazinyl, 1,2,3, 5-tetrahydrooxazolyl or 4H- [1,4]Thiazinyl, or it may be benzo-fused, such as, but not limited to, dihydroisoquinolinyl. When the 3-14 membered heterocyclic group is linked to other groups to form the compound of the present invention, the carbon atom on the 3-14 membered heterocyclic group may be linked to other groups, or the heterocyclic atom on the 3-14 membered heterocyclic ring may be linked to other groups. For example, when the 3-14 membered heterocyclic group is selected from piperazinyl, it may be that the nitrogen atom on the piperazinyl group is attached to other groups. Or, when the 3-14 membered heterocyclic group is selected from piperidyl, it may be a nitrogen atom on the piperidyl ring and a carbon atom in the para position thereofTo other groups.
The term "spiro" refers to a ring system in which two rings share 1 ring-forming atom.
The term "fused ring" refers to a ring system in which two rings share 2 ring atoms.
The term "bridged ring" refers to a ring system in which two rings share more than 3 ring members.
The term "halogen" means fluorine, chlorine, bromine and iodine.
"halo" refers to substitution with one or more halogens.
Those skilled in the art will appreciate that the compounds of formula (I) may exist in various pharmaceutically acceptable salt forms. If these compounds have a basic center, they may form acid addition salts; if these compounds have an acidic center, they may form base addition salts; these compounds may also form internal salts if they contain both acidic (e.g., carboxyl) and basic (e.g., amino) centers.
The compounds of the invention may exist in the form of solvates (e.g. hydrates) wherein the compounds of the invention comprise a polar solvent as a structural element of the compound lattice, in particular, for example, water, methanol or ethanol. The polar solvent, in particular water, may be present in stoichiometric or non-stoichiometric amounts.
Depending on its molecular structure, the compound of the invention may be chiral and thus various enantiomeric forms may exist. These compounds may thus be present in racemic or optically active form. The compounds of the present invention encompass isomers or mixtures, racemates thereof wherein each chiral carbon is in the R or S configuration. The compounds of the invention or intermediates thereof may be isolated as enantiomer compounds by chemical or physical methods well known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g.N-benzoylproline or N-benzenesulfonylproline) or various optically active camphorsulfonic acids in R and S form. The chromatographic resolution can also advantageously be carried out with the aid of optically active resolving agents, such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chiral derivatized methacrylate polymers, immobilized on silica. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example hexane/isopropanol/acetonitrile.
The corresponding stable isomer may be isolated according to known methods, for example by extraction, filtration or column chromatography.
The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses or primates, most preferably humans.
The term "therapeutically effective amount" refers to that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response that is being sought by a researcher, veterinarian, medical doctor or other clinician in a tissue, system, animal, individual or human, which includes one or more of the following: (1) prevention of disease: for example, preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not experienced or developed a pathology or symptomatology of the disease. (2) inhibition of disease: for example, inhibiting a disease, disorder or condition (i.e., preventing further development of pathology and/or symptoms) in an individual experiencing or presenting with the pathology or symptoms of the disease, disorder or condition. (3) alleviation of disease: for example, alleviating a disease, disorder or condition (i.e., reversing the pathology and/or symptoms) in an individual experiencing or presenting with the pathology or symptoms of the disease, disorder or condition.
Detailed Description
The technical scheme of the invention will be further described in detail below with reference to specific embodiments. It is to be understood that the following examples are illustrative only and are not to be construed as limiting the scope of the invention. All techniques implemented based on the above description of the invention are intended to be included within the scope of the invention.
Unless otherwise indicated, the starting materials and reagents used in the following examples were either commercially available or may be prepared by known methods.
Example 1
N- [2- (4, 4-Dihaloperidol-1-yl) -6-methylpyrimidin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 1)
First step Synthesis of methyl 4-bromo-2- (spiro [2.5] oct-5-en-6-yl) benzoate (Compound 1-3):
to a solution of methyl 4-bromo-2-iodobenzoate (compound 1-1, 700mg,2.053mmol,1 eq), sodium carbonate (326.41 mg,3.079mmol,1.5 eq) and 4, 5-tetramethyl-2- { spiro [2.5] oct-5-en-6-yl } -1,3, 2-dioxaborolan (compound 1-2, 480.74mg,2.053mmol,1 eq) in 1, 4-dioxane (8 mL) and water (1.6 mL) was added at room temperature [1, 1-bis (diphenylphosphine) ferrocene ] palladium dichloride dichloromethane complex (167.25 mg,0.205mmol,0.1 eq). The reaction solution was heated to 80℃and stirred for 1 hour. The reaction was cooled to room temperature, the reaction was then filtered, the filter cake was washed with ethyl acetate (3X 5 mL), the filtrate was diluted with water (30 mL), and the mixture was extracted with ethyl acetate (3X 30 mL). The organic phases were combined, backwashed with saturated brine (2X 40 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue obtained was purified with reverse phase column under the following conditions: chromatographic column specification, C18; mobile phase: acetonitrile and water, gradient 10% to 70%; elution time: for 10min; methyl 4-bromo-2- (spiro [2.5] oct-5-en-6-yl) benzoate (compound 1-3, 320mg, 48.52%) was obtained.
MS(ESI,m/z)：320.95[M+H] + ,RT(min):1.492
Synthesis of second step 4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzoic acid (Compounds 1-4):
to a solution of methyl 4-bromo-2- (spiro [2.5] oct-5-en-6-yl) benzoate (compound 1-3, 300mg,0.934mmol,1 eq) in methanol (2 mL), tetrahydrofuran (2 mL) and water (2 mL) was added lithium hydroxide (111.84 mg, 4.640 mmol,5 eq) at room temperature. The reaction was stirred at room temperature overnight. The reaction mixture was diluted with water (15 mL), the mixture was extracted with ethyl acetate (1X 10 mL), the aqueous phase was collected and acidified to pH 4-5 with 1 mol/L of dilute hydrochloric acid, and the aqueous phase was extracted with ethyl acetate (3X 15 mL). The organic phase was collected and backwashed with saturated saline (2X 20 mL) and dried over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure to give 4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzoic acid (compound 1-4, 250mg, 87.14%).
MS(ESI,m/z):306.90[M+H]+,RT(min):1.323
Step three Synthesis of 4-bromo-N- (2- (4, 4-difluoropiperidin-1-yl) -6-methylpyrimidin-4-yl) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (Compound 1-6)
To a solution of 4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzoic acid (compound 1-4, 100mg,0.326mmol,1 eq), 2- (4, 4-difluoropiperidin-1-yl) -6-methylpyridin-4-amine (compound 1-5, 89.16mg,0.391mmol,1.2 eq) and tetramethyl chloroformyl amidine hexafluorophosphate (182.68 mg,0.652mmol,2 eq) in N, N-dimethylformamide (3 mL) at room temperature was added 1-methyl-1H-imidazole (106.91 mg,1.304mmol,4 eq) and the reaction was warmed to 80℃and stirred overnight. The reaction was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (3X 20 mL). The organic phases were combined, backwashed with water (3X 20 mL) and dried over anhydrous sodium sulfate. Filtration, concentration of the filtrate under reduced pressure, and purification of the resulting residue by preparative chromatography plate (ethyl acetate: petroleum ether=1:5) gave 4-bromo-N- (2- (4, 4-difluoropiperidin-1-yl) -6-methylpyridin-4-yl) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (compound 1-6, 40mg, 21.77%).
MS(ESI,m/z)：517.30[M+H] + ,RT(min):1.477
Synthesis of N- [2- (4, 4-difluoropiperidin-1-yl) -6-methylpyrimidin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 1)
Palladium acetate (1.74 mg,0.008mmol,0.1 eq) and 2-di-tert-butylphosphino-2 ',4',6' -triisopropylbiphenyl (6.57 mg,0.015mmol,0.2 eq) were added to a solution of 4-bromo-N- [2- (4, 4-difluoropiperidin-1-yl) -6-methylpyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 1-6, 40mg,0.077mmol,1 eq), 2-hydroxyethanesulfonamide (compound 1-7, 19.35mg,0.154mmol,2 eq) and sodium tert-butoxide (22.29 mg,0.231mmol,3 eq) in N, N-dimethylformamide (2 mL) at room temperature under nitrogen atmosphere, and the reaction was stirred for 1 hour at 140 ℃. The reaction was cooled to room temperature, the reaction was filtered, and the filter cake was washed with ethyl acetate (3X 3 mL). The filtrate was diluted with ethyl acetate (15 mL), the mixture was washed with water (3X 10 mL), and the organic phase was dried over anhydrous sodium sulfate. Filtering, concentrating the filtrate under reduced pressure, and purifying the obtained crude product by using a prepared high-pressure liquid phase under the following conditions: chromatographic column specification: kineex EVO C18 Column,30 x 150,5 μm; mobile phase a: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL/min; elution gradient: 20% b to 75% b; elution time: 8min, N- [2- (4, 4-difluoropiperidin-1-yl) -6-methylpyrimidin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 1, 15.5mg, 32.18%) was obtained.
MS(ESI,m/z):561.90[M+H] + ,RT(min):1.766
1 H NMR(400MHz,DMSO-d6)δ10.08(s,2H),7.49(d,J＝8.4Hz,1H),7.21-7.14(m,2H),7.05(d,J＝2.3Hz,1H),5.73(s,1H),4.95(s,1H),3.83(s,4H),3.76(t,J＝6.6Hz,2H),3.29(d,J＝6.7Hz,2H),2.29(s,5H),2.03-1.84(m,6H),1.41(t,J＝6.0Hz,2H),0.31-0.18(m,4H).
Example 2
N- [2- (4, 4-difluoropiperidin-1-yl) -6-methylpyrimidin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-6-yl } benzamide (compound 2)
To a solution of N- [2- (4, 4-difluoropiperidin-1-yl) -6-methylpyrimidin-4-yl ] -4- (2-hydroxyethanesulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (10 mg,0.018mmol,1 eq) in ethyl acetate (1 mL) at room temperature under a hydrogen atmosphere was added platinum dioxide (0.40 mg,0.002mmol,0.10 eq), pressurized to 4MPa and stirred at room temperature for 5 days. The reaction solution was filtered, and the cake was washed with ethyl acetate (3X 3 mL), and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (Column format: XBridge Shield RP OBD Column, 30X 150mm,5 μm, N; mobile phase A: water (10 mmol/L) ammonium bicarbonate, mobile phase B: acetonitrile; flow rate: 60mL/min; elution gradient: 38%B to 70%B in 10min,70%B; detection wavelength: UV 220nm; retention time (min): 8.82), yielding N- [2- (4, 4-difluoropiperidin-1-yl) -6-methylpyridin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] octane-6-yl } benzamide (0.84 mg, 8.24%).
MS(ES,m/z):564.40[M+H] + ,RT(min):1.816
1 H NMR:(400MHz,CDCl 3 )δ7.86(s,1H),7.47-7.43(m,2H),7.28(s,1H),7.20-7.17(m,1H),6.78(s,1H),4.14(t,J＝5.2Hz,2H),3.93(s,4H),3.37-3.26(m,2H),3.05(t,J＝12.0Hz,1H),2.41(s,3H),2.01-1.94(m,4H),1.93-1.79(m,4H),1.64-1.62(m,2H),1.26(s,1H),0.92(d,J＝13.2Hz,2H),0.33-0.30(m,2H),0.28-0.24(m,2H).
Example 3
N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 3)
First step synthesis of 2- (4, 4-difluoropiperidin-1-yl) -6-nitropyridine (compound 3-3):
to a solution of 2-chloro-6-nitropyridine (1 g,6.31mmol,1 eq), 4-difluoropiperidine hydrochloride (1.15 g,9.46mmol,1.5 eq) and cesium carbonate (6.17 g,18.92mmol,3 eq) in toluene (10 mL) were added 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine (392.76 mg,0.63mmol,0.1 eq), palladium acetate (283.22 mg,1.26mmol,0.2 eq) at room temperature under nitrogen, the reaction was warmed to 100℃and stirred for 16 hours. The reaction was cooled to room temperature, diluted with water (50 mL) and the reaction mixture was extracted with ethyl acetate (3X 50 mL). The organic phases were combined, backwashed with saturated saline (1X 60 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (10:1) to give 2- (4, 4-difluoropiperidin-1-yl) -6-nitropyridine (480 mg, 63.88%).
MS(ESI,m/z):244.00[M+H] + ,RT(min):1.298
Second step Synthesis of 6- (4, 4-difluoropiperidin-1-yl) pyridin-2-amine (Compound 3-4):
to a mixed solution of 2- (4, 4-difluoropiperidin-1-yl) -6-nitropyridine (480 mg,3.62mmol,1 eq) in ethanol (10 mL) and water (1 mL) under nitrogen protection at room temperature were added iron powder (1.01 g,18.09mmol,5 eq) and ammonium chloride (967.69 mg,18.09mmol,5 eq), the reaction solution was warmed to 60℃and stirred for one hour, and the liquid was monitored to end the reaction. Cooled to room temperature and diluted with water (30 mL). The mixture was filtered, and the filter cake was washed with ethyl acetate (3X 20 mL). The filtrate was extracted with ethyl acetate (3X 50 mL). The organic phases were combined, backwashed with saturated saline (1X 60 mL) and dried over anhydrous sodium sulfate. Filtration and concentration of the filtrate under reduced pressure gave 6- (4, 4-difluoropiperidin-1-yl) pyridin-2-amine (600 mg, 77.77%).
MS(ESI,m/z):214.05[M+H] + ,RT(min):0.740
Step three synthesis of 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 3-5):
to a solution of 6- (4, 4-difluoropiperidin-1-yl) pyridin-2-amine (90 mg,0.21mmol,1 eq) in N, N-dimethylformamide (1 mL) was added N, N, N ', N' -tetramethyl chloroformyl amidine hexafluorophosphate (236.30 mg,0.41mmol,2 eq), N-methylimidazole (172.87 mg,1.03mmol,5 eq), 4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzoic acid (253.54 mg,0.41mmol,2 eq) at room temperature under nitrogen, and the reaction mixture was warmed to 80℃and stirred for one hour. The reaction solution was cooled to room temperature, and diluted with water (10 mL). The reaction mixture was extracted with ethyl acetate (3X 5 mL). The organic phases were combined, backwashed with saturated saline (3X 5 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (5:1) to give 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (90 mg, 42.44%).
MS(ESI,m/z):501.95[M+H] + ,RT(min):1.497
Fourth step Synthesis of N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 3):
To a solution of 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (75 mg,0.15mmol,1 eq) in 1, 4-dioxane (2 mL) under nitrogen, dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine (15.97 mg,0.03mmol,0.2 eq), (methanesulfonic acid { dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (13.71 mg,0.015mmol,0.1 eq), cesium carbonate (145.92 mg,0.45mmol,3 eq), 2-hydroxyethanesulfonamide (37.30 mg,0.30 mmol), sodium bicarbonate (20 mL) was added to a solution of 1, 20m aqueous solution (20 m) of water was diluted solution, and the mixture was concentrated to a saturated aqueous solution (20 m, 150 m of water solution was saturated with water solution (20 m, 20 m) and saturated aqueous solution (20 m) was cooled under reflux conditions at room temperature, mobile phase B, acetonitrile; the flow rate is 60mL/min; gradient 35%B to 65%B in 8min,65%B; the wavelength is 220nm; RT (min.) 7.32) to give N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (22.9 mg, 28.01%).
MS(ESI,m/z):547.00[M+H] + ,RT(min):1.706
1 H NMR(400MHz,DMSO-d6)δ9.98(s,1H),9.68(s,1H),7.61–7.50(m,2H),7.41(s,1H),7.21–7.14(m,1H),7.05(d,J＝2.2Hz,1H),6.66(d,J＝8.3Hz,1H),5.78(s,1H),4.95(s,1H),3.76(t,J＝6.6Hz,2H),3.66(s,4H),3.29(d,J＝6.7Hz,2H),2.27(s,2H),1.96(d,J＝15.8Hz,6H),1.41(s,2H),0.24(d,J＝2.6Hz,4H).
Example 4
N- [3- (4, 4-difluoropiperidin-1-yl) phenyl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 4)
First step synthesis of 4, 4-difluoro-1- (3-nitrophenyl) piperidine (Compound 4-2):
to a solution of 3-nitrochlorobenzene (2 g,12.694mmol,1 eq) in toluene (20 mL) was added 4, 4-difluoropiperidine hydrochloride (2.31 g,19.041mmol,1.5 eq), cesium carbonate (12.41 g,38.082mmol,3 eq), 1-binaphthyl-2, 2-diphenylphosphine (1580.93 mg,2.539mmol,0.2 eq), palladium acetate (285.00 mg, 1.399 mmol,0.1 eq) at room temperature under nitrogen, and the reaction mixture was warmed to 90℃and stirred for 1 hour. The reaction was cooled to room temperature and quenched by the addition of water (80 mL). The reaction mixture was extracted with ethyl acetate (3X 50 mL). The organic phases were combined, backwashed with saturated saline (1X 80 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (10:1) to give 4, 4-difluoro-1- (3-nitrophenyl) piperidine (2.92 g, 94.96%).
MS(ESI,m/z):243.00[M+H] + ,RT(min):1.242
Second step Synthesis of 3- (4, 4-difluoropiperidin-1-yl) aniline (Compound 4-3):
to a solution of 4, 4-difluoro-1- (3-nitrophenyl) piperidine (500 mg,2.06mmol,1 eq) in ethanol (5 mL) and water (1 mL) was added iron powder (576.37 mg,10.321mmol,5.00 eq) and ammonium chloride (552.07 mg,10.32mmol,5 eq) at room temperature, the reaction was warmed to 60℃and stirred for 1 hour. The reaction solution was cooled to room temperature, and the reaction mixture was extracted with ethyl acetate (3X 40 mL). The organic phases were combined, backwashed with saturated brine (1X 40 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (10:1) to give 3- (4, 4-difluoropiperidin-1-yl) aniline (400 mg, 91.32%).
MS(ESI,m/z):213.30[M+H] + ,RT(min):0.590
Step three Synthesis of 4-bromo-N- [3- (4, 4-difluoropiperidin-1-yl) phenyl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 4-4):
to a solution of 3- (4, 4-difluoropiperidin-1-yl) aniline (20 mg,0.094mmol,1 eq) in N, N-dimethylformamide (1 mL) was added 4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzoic acid (17.37 mg,0.056mmol,0.6 eq), tetramethyl chlorourea hexafluorophosphate (2 mg, 0.0070 mmol,0.08 eq), N-methylimidazole (5 mg,0.061mmol,0.65 eq) at room temperature under nitrogen atmosphere, and the reaction mixture was warmed to 80℃and stirred for 12 hours. The reaction solution was cooled to room temperature, diluted with water (10 mL), and the reaction mixture was extracted with ethyl acetate (3X 10 mL). The organic phases were combined, backwashed with saturated saline solution (1X 10 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (10:1) to give 4-bromo-N- [3- (4, 4-difluoropiperidin-1-yl) phenyl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (25 mg, 53.19%).
MS(ESI,m/z):501.00[M+H] + ,RT(min):1.281
Fourth step Synthesis of N- [3- (4, 4-difluoropiperidin-1-yl) phenyl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 4):
to a solution of 4-bromo-N- [3- (4, 4-difluoropiperidin-1-yl) phenyl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (25 mg,0.05mmol,1 eq) in N, N-dimethylformamide (0.5 mL) was added 2-hydroxyethanesulfonamide (7.50 mg,0.06mmol,1.2 eq), palladium acetate (1.1 mg,0.005mmol,0.1 eq), 2-di-tert-butylphosphino-2 ',4',6' -triisopropylbiphenyl (4.25 mg,0.01mmol,0.2 eq), sodium tert-butoxide (14.4 mg,0.15mmol,3 eq) at room temperature under nitrogen. The reaction solution was heated to 140℃and stirred for 1 hour. The reaction solution was cooled to room temperature and diluted with water (10 mL), and the reaction mixture was extracted with ethyl acetate (3X 10 mL). The organic phases were combined, backwashed with saturated saline solution (1X 10 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (Column: kinetex EVO C18 Column, 30X 150,5 μm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL/min; elution gradient: 35%B to 60%B in 8min,60%B; detection wavelength: 220nm; RT1 (min): 7.48). N- [3- (4, 4-difluoropiperidin-1-yl) phenyl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (1.3 mg, 23.73%) was obtained.
MS(ESI,m/z):545.90[M+H] + ,RT(min):1.603
1 H NMR(400MHz,DMSO-d 6 )δ9.78(s,1H),7.40(d,J＝8.2Hz,1H),7.35(s,1H),7.14(d,J＝8.2Hz,3H),7.06(s,1H),6.72(d,J＝7.6Hz,1H),5.71(s,1H),4.96(s,1H),3.75(t,J＝6.7Hz,2H),3.29(s,6H),2.31(s,2H),2.12–2.00(m,4H),1.92(s,2H),1.37(t,J＝6.0Hz,2H),0.22(d,J＝3.0Hz,2H),0.19(d,J＝3.0Hz,2H).
Example 5
N- [3- (4, 4-difluoropiperidin-1-yl) -5-methylphenyl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 5)
First step synthesis of 4, 4-difluoro-1- (3-methyl-5-nitrophenyl) piperidine (compound 5-2):
to a solution of 1, 4-dioxane (10 mL) of 1-chloro-3-methyl-5-nitrobenzene (1.0 g, 5.8238 mmol,1.0 eq), 4-difluoropiperidine hydrochloride (1.10 g,6.994mmol,1.2 eq), sodium t-butoxide (1.68 g, 17.284 mmol,3.0 eq) was added tris (dibenzylideneacetone) dipalladium (0.53 g,0.583mmol,0.1 eq), 4, 5-bis (diphenylphosphonic acid) -9.9-dimethylethane (0.67 g,1.166mmol,0.2 eq) at room temperature under nitrogen. The reaction solution was warmed to 110 degrees celsius and stirred for 16 hours. The reaction solution was cooled to room temperature, and diluted with water (40 mL). The reaction mixture was extracted with ethyl acetate (3X 60 mL). The organic phases were combined, backwashed with saturated saline (1X 80 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (9:1) to give 4, 4-difluoro-1- (3-methyl-5-nitrophenyl) piperidine (1.1 g, 73.65%).
MS(ESI,m/z):257.30[M+H] + ,RT(min):1.205.
Second step Synthesis of 3- (4, 4-difluoropiperidin-1-yl) -5-methylaniline (Compound 5-3):
To a solution of 4, 4-difluoro-1- (3-methyl-5-nitrophenyl) piperidine (500 mg,1.951mmol,1.0 eq) in ethanol (5 mL) and water (1 mL) was added iron powder (653.79 mg,11.706mmol,6.0 eq) and ammonium chloride (313.11 mg,5.853mmol,3.0 eq) at room temperature under nitrogen. The reaction solution was warmed to 60℃and stirred for 1 hour. The reaction solution was cooled to room temperature, filtered, and the filter cake was washed with ethyl acetate (3X 10 mL). The filtrate was collected and diluted with water (40 mL), extracted with ethyl acetate (3X 30 mL). The organic phases were combined, backwashed with saturated brine (1X 40 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was chromatographed on silica gel, petroleum ether/ethyl acetate (9:1) to give 3- (4, 4-difluoropiperidin-1-yl) -5-methylaniline (410 mg, 92.87%).
MS(ESI,m/z):227.00[M+H] + ,RT(min):0.681.
Step three synthesis of 4-bromo-N- [3- (4, 4-difluoropiperidin-1-yl) -5-methylphenyl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 5-4):
to a solution of 4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzoic acid (20 mg,0.065mmol,1.0 eq), 3- (4, 4-difluoropiperidin-1-yl) -5-methylaniline (17.68 mg,0.078mmol,1.2 eq) in N, N-dimethylformamide (1 mL) was added tetramethyl chloroformamide hexafluorophosphate (36.54 mg,0.130mmol,2.0 eq) and 1-methylimidazole (53.46 mg,0.650mmol,10.0 eq) at room temperature under nitrogen. The reaction solution was warmed to 80 degrees celsius and stirred for 2 hours. The reaction solution was cooled to room temperature and diluted with water (10 mL), and the mixture was extracted with ethyl acetate (3X 30 mL). The organic phases were combined, backwashed with saturated saline (1X 20 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was chromatographed on silica gel, petroleum ether/ethyl acetate (9:1) to give 4-bromo-N- [3- (4, 4-difluoropiperidin-1-yl) -5-methylphenyl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (30 mg, 89.40%).
MS(ESI,m/z):514.95[M+H] + ,RT(min):1.319.
Fourth step Synthesis of N- [3- (4, 4-difluoropiperidin-1-yl) -5-methylphenyl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 5):
palladium acetate (1.09 mg,0.005mmol,0.1 eq) and 2-di-tert-butyl phosphino-2 ',4',6' -triisopropylbiphenyl (4.12 mg,0.010mmol,0.2 eq) were added to a solution of 4-bromo-N- [3- (4, 4-difluoropiperidin-1-yl) -5-methylphenyl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (25 mg,0.049mmol,1.0 eq), 2-hydroxyethanesulfonamide (6.07 mg,0.049mmol, 1.eq) and sodium tert-butoxide (13.98 mg,0.147mmol,3.0 eq) in N, N-dimethylformamide (1 mL) at room temperature under nitrogen. The liquid monitoring reaction is complete. The reaction solution was cooled to room temperature and diluted with water (10 mL), and the mixture was extracted with ethyl acetate (3X 10 mL). The organic phases were combined, backwashed with saturated saline (1X 10 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography (column XBridge Prep OBD C, 30X 150mm,5 μm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: ACN; flow rate: 60 ml/min; gradient: 42% B to 65% B,65% B in 8 min; wavelength: 220 nm; retention time (min): 7.62) to give N- [3- (4, 4-difluoropiperidin-1-yl) -5-methylphenyl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (3.12 mg, 11.49%).
MS(ESI,m/z):559.90[M+H] + ，RT(min):1.715.
1 H NMR:(400MHz,DMSO-d 6 )δ9.73(s,2H),7.41(d,J＝8.3Hz,1H),7.17(dd,J＝8.3,2.2Hz,1H),7.13(s,1H),7.09(d,J＝2.2Hz,1H),6.98(s,1H),6.56(s,1H),5.71(s,1H),4.97(s,1H),3.76(t,J＝6.6Hz,2H),3.29–3.26(m,6H),2.33(s,2H),2.22(s,3H),2.03(s,4H),1.92(s,2H),1.38(t,J＝6.0Hz,2H),0.24–0.23(m,2H),0.19–0.17(m,2H).
Example 6
N- (6- (4, 4-difluoropiperidin-1-yl) pyrazin-2-yl) -4- ((2-hydroxyethyl) sulfonylamino) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (compound 6)
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First step synthesis of 6- (4, 4-difluoropiperidin-1-yl) pyrazin-2-amine (compound 6-2):
to a solution of 6-bromopyrazine-2-amine (200 mg,1.156mmol,1 eq) in N, N-dimethylacetamide (2 mL) was added potassium carbonate (480 mg, 3.463 mmol,3 eq) and 4, 4-difluoropiperidine hydrochloride (272 mg, 1.284 mmol,1.5 eq) at room temperature under nitrogen. The reaction solution was heated to 120℃and stirred for 16 hours. The reaction mixture was cooled to room temperature and diluted with water (20 mL). The mixture was extracted with ethyl acetate (3×30 mL), the organic phases were combined, backwashed with saturated brine (2×30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (1:1) to give 6- (4, 4-difluoropiperidin-1-yl) pyrazin-2-amine (180 mg, 72.82%).
MS(ESI,m/z):215.35[M+H] + ,RT(min):0.649.
Second step Synthesis of 4-bromo-N- (6- (4, 4-difluoropiperidin-1-yl) pyrazin-2-yl) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (Compound 6-3):
to a solution of 6- (4, 4-difluoropiperidin-1-yl) pyrazin-2-amine (41.84 mg,0.196mmol,1.2 eq) 4-bromo-2- (spiro [2.5] oct-5-en-6-yl) benzoic acid (50 mg,0.163mmol,1.00 eq) in dichloromethane (1 mL) was added N, N' -tetramethyl chloroformidine hexafluorophosphate (182.68 mg,0.652mmol,4 eq), N-methylimidazole (133.64 mg,1.630mmol,10 eq) at room temperature under nitrogen. The reaction solution was heated to 80℃and stirred for 5 hours. The reaction mixture was cooled to room temperature, diluted with water (10 mL), extracted with ethyl acetate (2X 30 mL), the organic phases combined, backwashed with saturated brine (1X 30 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (1:1) to give 4-bromo-N- (6- (4, 4-difluoropiperidin-1-yl) pyrazin-2-yl) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (40 mg, 48.82%).
MS(ESI,m/z):503.30[M+H] + ,RT(min):1.395.
Step three synthesis of N- (6- (4, 4-difluoropiperidin-1-yl) pyrazin-2-yl) -4- ((2-hydroxyethyl) sulfonylamino) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (compound 6):
to a solution of 4-bromo-N- (6- (4, 4-difluoropiperidin-1-yl) pyrazin-2-yl) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (40 mg,0.079mmol,1 eq), cesium carbonate (77.67 mg,0.237mmol,3 eq) and 2-hydroxyethane-1-sulfonamide (11.93 mg,0.095mmol,1.2 eq) in N, N-dimethylformamide (1 mL) was added (methanesulfonic acid { dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (7.30 mg,0.008mmol,0.1 eq) at room temperature under nitrogen, dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine (8.50 mg,0.016mmol,0.2 eq.) the reaction mixture was warmed to 100 ℃, stirred for 1 hour, the reaction mixture was cooled to room temperature, diluted with water (10 mL), extracted with ethyl acetate (3 x 30 mL), the organic phases were combined, backwashed with saturated brine (2 x 30 mL), dried over anhydrous sodium sulfate, the mixture was filtered, the filtrate was concentrated under reduced pressure, the crude product was purified by preparative high performance liquid phase, n- (6- (4, 4-difluoropiperidin-1-yl) pyrazin-2-yl) -4- ((2-hydroxyethyl) sulfonylamino) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (9.67 mg, 21.93%) was obtained.
MS(ESI,m/z):548.35[M+H] + ,RT(min):1.369.
1 H NMR(400MHz,DMSO-d 6 )δ10.08(s,1H),9.98(s,1H),8.58(s,1H),8.13(s,1H),7.51(d,J＝8.3Hz,1H),7.18(dd,J＝8.4,2.2Hz,1H),7.06(d,J＝2.2Hz,1H),5.73(s,1H),4.95(s,1H),3.76(t,J＝6.6Hz,2H),3.71(s,4H),3.31(s,2H),2.29(s,2H),1.99(s,4H),1.95(s,2H),1.40(t,J＝5.9Hz,2H),0.23(d,J＝12.1Hz,4H).
Example 7
N- [2- (4, 4-difluoropiperidin-1-yl) pyridin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- (spiro [2.5] oct-5-en-6-yl benzamide (compound 8)
First step synthesis of 2- (4, 4-difluoropiperidin-1-yl) pyridin-4-amine (compound 8-2):
to a solution of 2-chloropyridin-4-amine (500 mg,3.889mmol,1 eq) in N-methylpyrrolidone (5 mL) was added N, N-diisopropylethylamine (1508.02 mg,11.667mmol,3 eq) at room temperature under nitrogen, 4-difluoropiperidine hydrochloride (706.66 mg,5.833mmol,1.5 eq) was added. The reaction system was subjected to microwave reaction at 160℃for 2 hours. The reaction was cooled to room temperature and quenched by the addition of water (5 mL). The reaction mixture was extracted with ethyl acetate (3X 30 mL). The organic phases were combined, backwashed with saturated saline (3X 30 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (5:1) to give 2- (4, 4-difluoropiperidin-1-yl) pyridin-4-amine (300 mg, 36.17%).
MS:(ESI,m/z):214.05[M+H] + ,RT(min):0.820
Second step Synthesis of 4-bromo-N- (2- (4, 4-difluoropiperidin-1-yl) pyridin-4-yl) -2-spiro [2.5] oct-5-en-6-yl) benzamide (Compound 8-3):
n, N, N ', N' -tetramethyl-chloroformyl-hexafiuorophosphate (526.34 mg,1.876mmol,4 eq), 4-bromo-2- (spiro [2.5] oct-5-en-6-ylbenzoic acid (172.87 mg,0.563mmol,1.2 eq), N-methylimidazole (385.05 mg, 4.460 mmol,10 eq) were added to a solution of 2- (4, 4-difluoropiperidin-1-yl) pyridin-4-amine (100 mg,0.469mmol,1 eq) in methylene chloride (2 mL) at room temperature, the reaction mixture was stirred for 2 hours until it was cooled to room temperature, water (10 mL) was added, the organic phase was combined, the mixture was backwashed with saturated brine (1X 20 mL), and the resulting mixture was dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography, petroleum ether/ethyl acetate (10:1) to give N-methyl imidazole (385.05 mg, 4.460 mmol,10 eq) and 4-bromo-4- (4-5-benzoyl) -2-4-piperidinyl-4-amine (3.20 mg).
MS:(ESI,m/z):502.35[M+H] + ,RT(min):1.368
Step three synthesis of N- [2- (4, 4-difluoropiperidin-1-yl) pyridin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- (spiro [2.5] oct-5-en-6-yl benzamide (compound 8):
to 4-bromo-N- (2- (4, 4-difluoropiperidin-1-yl) pyridin-4-yl) -2-spiro [2.5] at room temperature under nitrogen]Oct-5-en-6-yl) benzoylTo a solution of 1,4 dioxane (1 mL) of cesium carbonate (121.59 mg,0.372mmol,3 eq) in 2-hydroxyethane-1-sulfonamide (23.35 mg,0.186mmol,1.5 eq) was added dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl]-2-yl) phosphanes(6.65mg,0.012mmol,0.1 eq), (methanesulfonic acid { dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl)]-2-yl) phosphine alkane } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (22.85 mg,0.025mmol,0.2 eq). The reaction solution was heated to 100℃and stirred for 1 hour. The reaction was cooled to room temperature and quenched by the addition of water (3 mL). The reaction mixture was extracted with ethyl acetate (3X 20 mL). The organic phases were combined, backwashed with saturated saline (1X 20 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. Purifying the crude product by high performance liquid chromatography under the following conditions of XBridge Prep OBD C, 30 x 150mm and 5 μm; mobile phase a: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60ml/min; elution gradient: 46%B to 73%B in 8min; detection wavelength: 254nm; retention time (min): 6.72. to obtain N- [2- (4, 4-difluoropiperidin-1-yl) pyridin-4-yl ]-4- ((2-hydroxyethyl) sulphonylamino) -2- (spiro [ 2.5)]Oct-5-en-6-yl benzamide (11.18 mg, 16.44%).
MS:(ESI,m/z):547.00[M+H] + ,RT(min):1.611
1 H NMR:(400MHz,DMSO-d 6 )δ10.15(s,1H),9.99(s,1H),8.01(d,J＝5.6Hz,1H),7.44(d,J＝8.4Hz,1H),7.25(s,1H),7.18(dd,J＝8.3,2.2Hz,1H),7.10(d,J＝2.1Hz,1H),6.94(d,J＝5.4Hz,1H),5.69(s,1H),4.98(s,1H),3.76(s,2H),3.62(t,J＝5.8Hz,4H),3.28(d,J＝6.6Hz,2H),2.32(s,2H),1.98(dt,J＝14.6,8.4Hz,4H),1.90(s,2H),1.37(t,J＝6.0Hz,2H),0.22(d,J＝3.6Hz,2H),0.17(d,J＝3.8Hz,2H).
Example 8
N- (2- (3, 3-Difluoroazetidin-1-yl) -6-methylpyrimidin-4-yl) -4- ((2-hydroxyethyl) sulfonylamino) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (Compound 29)
First step synthesis of 2- (3, 3-difluoroazetidin-1-yl) -6-methylpyrimidin-4-amine (compound 29-3):
2-chloro-6-methylpyrimidin-4-amine (1.00 g,6.965mmol,1 eq), N, N-diisopropylethylamine (2.70 g,20.895mmol,3 eq) and N-methylpyrrolidone (20 mL) of 3, 3-difluoroazetidine hydrochloride (1.35 g,10.447mmol,1.5 eq) were reacted under stirring at 140℃for 16 hours under nitrogen. The reaction mixture was cooled to room temperature and diluted with water (50 mL). The mixture was extracted with ethyl acetate (3X 50 mL), the organic phases were combined, backwashed with water (3X 50 mL) and saturated brine (2X 50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue obtained was purified by reverse phase column chromatography under the following conditions: c18 column, mobile phase: water and acetonitrile, gradient 10% to 30% for 10 min, detector: UV 254nm, 2- (3, 3-difluoroazetidin-1-yl) -6-methylpyrimidin-4-amine (910 mg, 65.26%).
MS(ESI,m/z):201.25[M+H] + ,RT(min):0.627.
Second step Synthesis of 4-bromo-N- (2- (3, 3-difluoroazetidin-1-yl) -6-methylpyrimidin-4-yl) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (Compound 29-4):
To a solution of 2- (3, 3-difluoroazetidin-1-yl) -6-methylpyrimidin-4-amine (39.1 mg,0.196mmol,1.20 eq), 4-bromo-2- (spiro [2.5] oct-5-en-6-yl) benzoic acid (50 mg,0.163mmol,1.00 eq) in dichloromethane (1 mL) was added N, N, N ', N' -tetramethyl chloroformamidine hexafluorophosphate (182.68 mg,0.652mmol,4 eq), N-methylimidazole (133.64 mg,1.630mmol,10 eq) at room temperature under nitrogen. The reaction solution was stirred at room temperature for 16 hours. The reaction was extracted with dichloromethane (3X 10 mL). The organic phases were combined, backwashed with saturated saline (2X 10 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (5:1) to give 4-bromo-N- (2- (3, 3-difluoroazetidin-1-yl) -6-methylpyrimidin-4-yl) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (60 mg, 75.33%).
MS(ESI,m/z):489.25[M+H] + ,RT(min):1.455.
Step three synthesis of N- (2- (3, 3-difluoroazetidin-1-yl) -6-methylpyrimidin-4-yl) -4- ((2-hydroxyethyl) sulfonylamino) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (compound 29):
to a solution of 4-bromo-N- (2- (3, 3-difluoroazetidin-1-yl) -6-methylpyrimidin-4-yl) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (40 mg,0.082mmol,1 eq), cesium carbonate (79.9 mg,0.246mmol,3 eq) and 2-hydroxyethane-1-sulfonamide (12.27 mg,0.098mmol,1.2 eq) in N, N-dimethylformamide (1 mL), methanesulfonic acid { dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine } (2 '-methylamino 1,1' -biphenyl-2-yl) palladium (II) (7.51 mg,0.008mmol,0.1 eq), dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine (1, 1-2 eq), aqueous solution (1 mL) was added at room temperature, and the solution was concentrated to a solution of N, N-dimethylformamide (1 mL), and the solution was concentrated to a solution of saturated aqueous solution of liquid phase (1 mL), and saturated aqueous solution was cooled to room temperature (10 mL), and saturated aqueous solution was concentrated to room temperature (1 mL) by stirring; mobile phase A water (10 mmol/L ammonium bicarbonate), mobile phase B, acetonitrile; the flow rate is 60mL/min; elution gradient 35%B to 65%B in 10min,80%B; the detection wavelength is UV 220nm; retention time (min): 9.2) N- (2- (3, 3-difluoroazetidin-1-yl) -6-methylpyrimidin-4-yl) -4- ((2-hydroxyethyl) sulfonylamino) -2- (spiro [2.5] oct-5-en-6-yl) benzamide (16.63 mg, 37.60%) was obtained.
MS(ESI,m/z):534.45[M+H] + ,RT(min):1.456.
1 H NMR(400MHz,DMSO-d 6 )δ10.33(s,1H),9.98(s,1H),7.46(d,J＝8.4Hz,1H),7.37(s,1H),7.15(dd,J＝8.4,2.2Hz,1H),7.05(d,J＝2.3Hz,1H),5.71(s,1H),4.95(s,1H),4.38(t,J＝12.5Hz,4H),3.75(t,J＝6.6Hz,2H),3.31(s,2H),2.31(s,3H),2.29-2.24(m,2H),1.94(d,J＝3.6Hz,2H),1.41(t,J＝5.9Hz,2H),0.24(d,J＝12.7Hz,4H).
Example 9
N- [6- (4, 4-Dihaloperidol-1-yl) -4-methylpyridin-2-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 40)
First step synthesis of 6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-amine (compound 40-2):
to a solution of 6-chloro-4-methylpyridin-2-amine (500 mg,3.51mmol,1 eq) in tetrahydrofuran (10.0 mL) was added sodium tert-butoxide (2.02 g,21.042mmol,6 eq), 2-dicyclohexylphosphorus-2, 6-diisopropyloxy-1, 1-biphenyl (163.63 mg,0.351mmol,0.1 eq), 1- { [2',6' -bis (prop-2-yloxy) - [1,1' -biphenyl ] -2-yl ] dicyclohexyl- λ5-phosphono } -1-chloro-2H, 3H, 4H-benzo [ c ] 1-aza-2-palladium on cyclohexane (255.52 mg, 0.352 mmol,0.1 eq) and 4, 4-difluoropiperidine hydrochloride (849.51 mg, 7.51 mmol,2 eq) at room temperature under nitrogen. The reaction solution was heated to 100℃and stirred for one hour. The desired product was found in the liquid. The reaction was cooled to room temperature and quenched by the addition of water (20 mL). The reaction mixture was extracted with ethyl acetate (3X 40 mL). The organic phases were combined, backwashed with saturated saline (1X 50 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (5:1) to give 6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-amine (110 mg, 13.80%).
MS(ESI,m/z):228.40[M+H] + ,RT(min):0.584
Second step Synthesis of 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 40-3):
to a solution of 4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzoic acid (22 mg,0.072mmol,1.00 eq) in methylene chloride (2 mL) was added tetramethyl chloroformyl amidine hexafluorophosphate (80.38 mg,0.288mmol,4 eq) under nitrogen protection, and N-methylimidazole (58.8 mg,0.720mmol,10 eq) was added after stirring for 2 minutes at room temperature, and 6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-amine (24.41 mg,0.108mmol,1.50 eq) was added at room temperature. The reaction mixture was heated to 80℃and reacted for 1 hour. The reaction was cooled to room temperature and quenched by the addition of water (10 mL). The reaction mixture was extracted with ethyl acetate (3X 10 mL). The organic phases were combined, backwashed with saturated saline (1X 10 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (5:1) to give 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (35 mg, 94.63%).
MS(ESI,m/z):516.15[M+H] + ,RT(min):1.322
Step three synthesis of N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 40):
To a solution of 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (25 mg,0.048mmol,1 eq) in 1,4 dioxane (1 mL) was added dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine alkane (5.18 mg,0.010mmol,0.2 eq), methanesulfonic acid { dicyclohexyl [ 3-isopropoxy-2 ',4',6 '-triisopropyl- (1, 1' -biphenyl) -2-yl ] phosphine } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (4.45 mg,0.005mmol,0.1 eq), cesium carbonate (47.32 mg,0.144mmol,3 eq) and 2-hydroxyethanesulfonamide (12.09 mmol, 0.12 eq) at room temperature under nitrogen. The reaction solution was warmed to 120 ℃. The reaction was allowed to stand for one hour, cooled to room temperature, and quenched by the addition of water (10 mL). The reaction mixture was extracted with ethyl acetate (3X 10 mL). The organic phases were combined, backwashed with saturated saline (1X 10 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (column XBridge Shield RP OBD column, 30X 150mm,5 μm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; gradient: 40% B to 73% B,73% B over 10 min; wavelength: 220 nm; RT1 (min): 8.27). N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (7.98 mg, 29.40%) was obtained.
MS(ESI,m/z):561.15[M+H] + ,RT(min):1.847
1 H NMR(400MHz,DMSO-d 6 )δ9.97(s,1H),9.55(s,1H),7.54(d,J＝8.3Hz,1H),7.31(s,1H),7.17(d,J＝8.8Hz,1H),7.04(s,1H),6.51(d,J＝5.6Hz,1H),5.79(s,1H),4.94(s,1H),3.76(t,J＝6.6Hz,2H),3.65(s,4H),3.23(s,2H),2.37(s,2H),2.25(d,J＝10.3Hz,3H),1.99(s,2H),1.93(s,4H),1.44–1.38(m,2H),0.25(s,4H).
Example 10
N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -5- ((2-hydroxyethyl) sulfonylamino) -3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxamide (compound 41)
First step synthesis of 3-chloro-5-nitropyridine-2-carboxylic acid methyl ester (compound 41-2):
to methanol (10 mL) of 3-chloro-5-nitropyridine-2-carbonitrile (3.9 g,21.248mmol,1 eq) was added thionyl chloride (10 mL) at room temperature. After the addition was completed, the system was stirred at 50℃for 16 hours. The reaction was cooled to room temperature and quenched with water (100 mL). The reaction mixture was extracted with ethyl acetate (3X 100 mL). The organic phases were combined, backwashed with saturated saline (1X 100 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative chromatography plate, petroleum ether/ethyl acetate (10:1) to give methyl 3-chloro-5-nitropyridine-2-carboxylate (1.5 g, 32.60%).
MS(ESI,m/z):216.90[M+H] + ，RT(min):0.828.
Second step Synthesis of 5-nitro-3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxylic acid methyl ester (Compound 41-3):
to a solution of 3-chloro-5-nitropyridine-2-carboxylic acid methyl ester (100 mg, 0.460 mmol,1.00 eq), 4, 5-tetramethyl-2- { spiro [2.5] oct-5-en-6-yl } -1,3, 2-dioxaborolan (100 mg,0.427mmol,1.00 eq) in 1, 4-dioxane (1.7 mL) and water (0.4 mL) were added 1,1' -bis (diphenylphosphino) ferrocene-palladium (II) dichloride (31.25 mg,0.043mmol,0.1 eq) and sodium carbonate (146.81 mg, 1.3836 mmol,3.0 eq) at room temperature under nitrogen. The reaction solution was heated to 80℃and stirred for 1 hour. The reaction mixture was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3X 30 mL). The organic phases were combined, backwashed with saturated saline (1X 20 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative chromatography on a plate, petroleum ether/ethyl acetate (8:1) to give methyl 5-nitro-3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxylate (50 mg, 37.56%).
MS(ESI,m/z):289.00[M+H] + ,RT(min):1.110.
Synthesis of 5-nitro-3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxylic acid (Compound 41-4):
to methyl 5-nitro-3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxylate (40 mg,0.139mmol,1.0 eq) in methanol (2 mL) and water (0.4 mL) was added lithium hydroxide (33.23 mg,1.390mmol,10.0 eq) at room temperature. The reaction solution was warmed to 60℃and stirred for 1 hour. The reaction was cooled to room temperature and quenched with water. The reaction mixture was acidified to ph=6 with 1 mol per liter of hydrochloric acid. The reaction mixture was extracted with ethyl acetate (3X 20 mL). The organic phases were combined, backwashed with saturated saline (1X 30 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure to give 5-nitro-3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxylic acid (37 mg, 97.23%).
MS(ESI,m/z):274.90[MH] + ,RT(min):0.672.
Fourth step Synthesis of N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -5-nitro-3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxamide (compound 41-5):
to 5-nitro-3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxylic acid (30 mg,0.109mmol,1.0 eq), 6- (4, 4-difluoropiperidin-1-yl) pyridin-2-amine (25.66 mg,0.120mmol,1.1 eq), tetramethyl chloroformate hexafluorophosphate (92.07 mg,0.327mmol,3.0 eq) in dichloromethane (1 mL) was added N-methylimidazole (89.81 mg,1.090mmol,10.0 eq) at room temperature under nitrogen. After the addition was completed, stirring was continued at 80℃for 2 hours. The reaction mixture was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3X 20 mL). The organic phases were combined, backwashed with saturated saline (1X 20 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative chromatography on a plate, petroleum ether/ethyl acetate (3:2) to give N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -5-nitro-3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxamide (30 mg, 58.42%).
MS(ESI,m/z):470.25[M+H] + ,RT(min):1.379.
Fifth step Synthesis of 5-amino-N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxamide (compound 41-6):
to N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -5-nitro-3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxamide (30 mg,0.064mmol,1.0 eq) in ethanol (1 mL) and water (0.2 mL) was added iron powder (10.71 mg,0.192mmol,3.0 eq) and ammonium chloride (20.51 mg,0.384mmol,6.0 eq) at room temperature. The reaction solution was warmed to 60 degrees celsius and stirred for 1 hour. The reaction solution was cooled to room temperature, filtered, the filter cake was washed with ethyl acetate (3X 10 mL), and the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative chromatography on a plate, petroleum ether/ethyl acetate (5:1) to give 5-amino-N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxamide (30 mg, 90.80%).
MS(ESI,m/z):440.10[M+H] + ,RT(min):1.055.
Step six synthesis of ethyl 2- [ (6- { [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] carbamoyl } -5- { spiro [2.5] oct-5-en-6-yl } pyridin-3-yl) sulfamoyl ] acetate (compound 41-8):
to a solution of 5-amino-N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxamide (30 mg,0.055mmol,1.0 eq) in acetonitrile (1 mL) was added ethyl 2-chlorosulfonylacetate (11.2 mg,0.061mmol,1.1 eq) and potassium carbonate (15.1 mg,0.110mmol,2.0 eq) at 0deg.C. The reaction was carried out at room temperature for 1 hour, and the reaction mixture was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3X 20 mL). The combined organic phases were backwashed with saturated saline (1X 20 mL) and dried over sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative chromatography on plates, petroleum ether/ethyl acetate (5:1) to give ethyl 2- [ (6- { [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] carbamoyl } -5- { spiro [2.5] oct-5-en-6-yl } pyridin-3-yl) sulfamoyl ] acetate (20 mg, 62.11%).
MS(ESI,m/z):590.10[M+H] + ,RT(min):1.196.
Seventh step Synthesis of N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -5- ((2-hydroxyethyl) sulfonylamino) -3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxamide (compound 41):
to a solution of ethyl 2- [ (6- { [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] carbamoyl } -5- { spiro [2.5] oct-5-en-6-yl } pyridin-3-yl) sulfamoyl ] acetate (20 mg,0.034mmol,1 eq) in tetrahydrofuran (2 mL) at-40℃was added dropwise lithium aluminum hydride (0.02 mL,2.5mol/L,1.2 eq). After the addition was completed, the system was stirred at room temperature for 1 hour. The desired product was found in the liquid. The reaction mixture was quenched with water at 0deg.C, the mixture was extracted with ethyl acetate (3X 20 mL), the organic phases were combined, backwashed with saturated brine (1X 30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure, and the crude product was purified by high performance liquid chromatography under the following conditions (column size: XBridge Shield RP OBD column, 30X 150mm,5 μm; mobile phase A: water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL/min; gradient: 28% B to 63% B,63% B over 8 min; wavelength: 220nm; retention time (min): 7.93). N- [6- (4, 4-difluoropiperidin-1-yl) pyridin-2-yl ] -5- ((2-hydroxyethyl) sulfonylamino) -3- { spiro [2.5] oct-5-en-6-yl } pyridine-2-carboxamide (3.3 mg, 17.77%) was obtained.
MS(ESI,m/z):547.95[M+H] + ,RT(min):1.487.
1 H NMR:(400MHz,DMSO-d 6 )δ10.38(s,1H),10.11(s,1H),8.37(s,1H),7.60(t,J＝8.0Hz,1H),7.48(d,J＝7.8Hz,1H),7.43(d,J＝2.5Hz,1H),6.69(d,J＝8.3Hz,1H),5.62(s,1H),4.96(s,1H),3.78(t,J＝6.2Hz,2H),3.68(s,4H),3.37(d,J＝12.5Hz,2H),2.30(s,2H),1.99(d,J＝15.1Hz,6H),1.50(d,J＝6.0Hz,2H),0.33(d,J＝3.4Hz,4H).
Example 11
N- [6- (4, 4-difluoropiperidin-1-yl) -5-fluoropyridin-2-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 42)
First step synthesis of 6- (4, 4-difluoropiperidin-1-yl) -5-fluoropyridin-2-amine (compound 42-2):
to a solution of 6-bromo-5-fluoropyridin-2-amine (300 mg,1.571mmol,1 eq) in N-methylpyrrolidone (3 mL) was added N, N-diisopropylethylamine (609.01 mg, 4.719 mmol,3 eq) and 4, 4-difluoropiperidine hydrochloride (495.04 mg,3.142mmol,2 eq) at room temperature under nitrogen. Heating to 200 ℃, and stirring by microwaves for reacting for one hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3X 20 mL). The organic phases were combined, backwashed with water (3X 10 mL) and saturated brine (3X 10 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure to give 6- (4, 4-difluoropiperidin-1-yl) -5-fluoropyridin-2-amine (150 mg, 41.32%).
MS(ESI,m/z):232.05[M+H] + ,RT(min):1.210
Second step Synthesis of 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) -5-fluoropyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 42-3):
to a solution of 4-bromo-2- (spiro [2.5] oct-5-en-6-yl) benzoic acid (55 mg,0.177mmol,1 eq) and 6- (4, 4-difluoropiperidin-1-yl) -5-fluoropyridin-2-amine (82 mg,0.355mmol,2 eq) in dichloromethane (1.5 mL) was added N, N' -tetramethyl chloroformidine hexafluorophosphate (199.01 mg,0.710mmol,4 eq) and N-methylimidazole (145.59 mg,1.775mmol,10 eq) at room temperature. The reaction solution was stirred at room temperature for 1 hour. The reaction was extracted with dichloromethane (3X 10 mL). The organic phases were combined, backwashed with saturated saline (1X 10 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column on silica gel, petroleum ether/ethyl acetate (10:1) to give 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) -5-fluoropyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (55 mg, 59.60%).
MS(ESI,m/z):520.11[M+H] + ,RT(min):1.559
Step three synthesis of N- [6- (4, 4-difluoropiperidin-1-yl) -5-fluoropyridin-2-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 42):
to a solution of 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) -5-fluoropyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (55 mg,0.106mmol,1 eq) in 1, 4-dioxane (2 mL) was added cesium carbonate (103.31 mg,0.318mmol,3 eq), 2-hydroxyethane-1-sulfonamide (20 mg,0.159mmol,1.5 eq), dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine alkane (11.3 mg,0.021mmol,0.2 eq), (methanesulfonic acid { dicyclohexyl (3-isopropoxy-2 ',1.5 eq) under nitrogen atmosphere at room temperature, 4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (9.71 mg,0.01 mmol,0.1 eq.) the reaction mixture was warmed to 100 ℃, the solution stirred for 1 hour, the reaction mixture was extracted with ethyl acetate (3X 20 mL), the organic phases were combined, backwashed with saturated brine (1X 20 mL), dried over anhydrous sodium sulfate, the resulting mixture was filtered, the filtrate concentrated under reduced pressure, and the crude product was purified by preparative high performance liquid phase under the following conditions (wavelength: 254nm/220nm, a chromatographic column XSelect CSH Fluoro Phenyl μm,30mm x 150mm, a mobile phase A water (0.1% formic acid), a mobile phase B acetonitrile, a flow rate of 60mL/min, a gradient of 45% B to 65% B within 10min, retention time (min): 10.08). N- [6- (4, 4-difluoropiperidin-1-yl) -5-fluoropyridin-2-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (6.03 mg, 10.10%) was obtained.
MS(ESI,m/z):565.40[M+H] + ,RT(min):1.666
1 H NMR(400MHz,CDCl 3 )δ8.78(s,1H),7.86(d,J＝8.4Hz,1H),7.81–7.75(m,1H),7.33(dd,J＝12.2,8.5Hz,1H),7.22(dd,J＝8.4,2.3Hz,1H),7.13(d,J＝2.3Hz,1H),6.90(s,1H),6.01–5.95(m,1H),4.13(t,J＝5.2Hz,2H),3.63(t,J＝5.8Hz,4H),3.35(t,J＝5.2Hz,2H),2.47(s,1H),2.33(q,J＝5.2Hz,2H),2.12(q,J＝3.9,3.3Hz,2H),2.05(dt,J＝13.6,5.7Hz,4H),1.50(t,J＝6.0Hz,2H),0.38–0.24(m,4H).
Example 12
N- [2- (4, 4-difluoropiperidin-1-yl) -3-fluoropyridin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 43)
First step synthesis of 2- (4, 4-difluoropiperidin-1-yl) -3-fluoropyridin-4-amine (compound 43-2):
to a solution of 2-chloro-3-fluoropyridin-4-amine (100 mg,0.68mmol,1 eq) in N-methylpyrrolidone (1 mL) was added 4, 4-difluoropiperidine hydrochloride (161 mg,1.02mmol,1.5 eq) and diisopropylethylamine (265 mg,2.05mmol,3 eq) at room temperature under nitrogen. The reaction solution is heated to 200 ℃ and subjected to microwave reaction for 3 hours. The reaction was cooled to room temperature, quenched with water, and the reaction mixture was extracted with ethyl acetate (3X 50 mL). The organic phases were combined, backwashed with water (3X 40 mL) and saturated brine (2X 60 mL) and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. 2- (4, 4-difluoropiperidin-1-yl) -3-fluoropyridin-4-amine (38 mg, 2.41%) was obtained.
MS(ESI,m/z):232.05[M+H] + ,RT(min):0.556.
Second step Synthesis of 4-bromo-N- [2- (4, 4-difluoropiperidin-1-yl) -3-fluoropyridin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 43-3):
to a solution of 2- (4, 4-difluoropiperidin-1-yl) -3-fluoropyridin-4-amine (30 mg,0.130mmol,1 eq) in dichloromethane (5 mL) was added 4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzoic acid (47.83 mg,0.156mmol,1.2 eq), tetramethyl chloroformidine hexafluorophosphate (47.83 mg,0.156mmol,1.2 eq), N-methylimidazole (106.53 mg,1.300mmol,10 eq) at room temperature under nitrogen. The reaction solution was heated to 80℃and stirred for 2 hours. The reaction mixture was cooled to room temperature, quenched with water, and the reaction mixture was extracted with dichloromethane (3X 30 mL). The organic phases were combined, backwashed with saturated saline (1X 30 mL) and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (3:1) to give 4-bromo-N- [2- (4, 4-difluoropiperidin-1-yl) -3-fluoropyridin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (45 mg, 66.65%).
MS(ESI,m/z):520.11[M+H] + ,RT(min):1.773
Step three synthesis of N- [2- (4, 4-difluoropiperidin-1-yl) -3-fluoropyridin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 43):
to a solution of 4-bromo-N- [2- (4, 4-difluoropiperidin-1-yl) -3-fluoropyridin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (40 mg,0.077mmol,1 eq) in N, N-dimethylformamide (5 mL) was added 2-hydroxyethanesulfonamide (19.24 mg,0.154mmol,2 eq), palladium acetate (1.73 mg,0.008mmol,0.1 eq), 2-di-tert-butyl phosphino-2 ',4',6' -triisopropylbiphenyl (6.53 mg,0.015mmol,0.2 eq), sodium tert-butoxide (22.16 mg,0.231mmol,3 eq) at room temperature under nitrogen. The reaction solution was heated to 140℃and stirred for 1 hour. The reaction mixture was cooled to room temperature, quenched with water, and the reaction mixture was extracted with ethyl acetate (3X 30 mL). The organic phases were combined, backwashed with saturated saline (1X 30 mL) and dried over anhydrous sodium sulfate. Filtering, and concentrating the filtrate under reduced pressure. The residue obtained was purified with preparative high performance liquid phase under the following conditions: column specification: xselect CSH C18 OBD,30 x 150mm,5 μm; mobile phase A water (0.1% formic acid), mobile phase B acetonitrile; the flow rate is 60mL/min; elution gradient 65%Bto 90%B in 8min,90%B; the detection wavelength is 220nm; retention time (min) 7.78. N- [2- (4, 4-difluoropiperidin-1-yl) -3-fluoropyridin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (2.3 mg, 5.25%) was obtained.
MS(ESI,m/z):564.95[M+H] + ,RT(min):1.613.
1 H NMR(400MHz,DMSO-d 6 )δ10.13-9.79(s,2H),8.00-7.87(d,J＝5.5Hz,1H),7.65-7.53(s,1H),7.54-7.42(d,J＝8.4Hz,1H),7.28-7.10(d,J＝8.5Hz,1H),7.10-6.94(s,1H),5.78-5.67(s,1H),5.05-4.86(s,1H),3.85-3.65(t,J＝6.6Hz,2H),3.58-3.32(m,4H),3.29-3.19(m,2H),2.39-2.26(s,2H),2.24-1.98(m,4H),1.98-1.76(s,2H),1.46-1.29(m,2H),0.37-0.22(d,J＝3.5Hz,2H),0.22-0.06(m,2H).
Example 13
N- [2- (4, 4-Difluorocyclohexyl) -6-methylpyrimidin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 44)
First step synthesis of 2- (4, 4-difluorocyclohex-1-en-1-yl) -6-methylpyrimidin-4-amine (compound 44-2):
to a mixed (20 mL,4/1, v/v) solution of 2-chloro-4-amino-6-methylpyrimidine (800 mg, 5.578mmol, 1 eq) in 1,4 dioxane and water was added 4, 4-difluorocyclohexene-1-boronic acid pinacol ester (1631.54 mg,6.686mmol,1.2 eq), potassium phosphate (2365.56 mg,11.144mmol,2 eq) and 1, 1-bis (diphenylphosphoryl) ferrocene palladium dichloride (454.69 mg,0.557mmol,0.1 eq) at room temperature under nitrogen. The reaction solution was heated to 80℃and stirred for 4 hours. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3X 50 mL). The organic phases were combined, backwashed with saturated saline (1X 50 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, ethyl acetate/petroleum ether (1:1) to give 2- (4, 4-difluorocyclohex-1-en-1-yl) -6-methylpyrimidin-4-amine (684.5 mg, 89.975%).
MS:(ESI,m/z):226.10[M+H] + ,RT(min):0.685.
Second step Synthesis of 2- (4, 4-difluorocyclohexane-1-yl) -6-methylpyrimidin-4-amine (Compound 44-3):
Palladium on carbon (68 mg,0.21mmol,0.69 eq) was added to a solution of 2- (4, 4-difluorocyclohex-1-en-1-yl) -6-methylpyridin-4-amine (684.5 mg,3.039mmol,1 eq) in ethanol (15 mL) under nitrogen at room temperature, hydrogen was substituted, and the reaction mixture was reacted at room temperature for 6 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give 2- (4, 4-difluorocyclohexyl) -6-methylpyrimidin-4-amine (620 mg, 90%).
MS:(ESI,m/z):228.20[M+H] + ,RT(min):0.737.
Step three synthesis of 4-bromo-N- [2- (4, 4-difluorocyclohexyl) -6-methylpyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 44-4):
to a solution of 2- (4, 4-difluorocyclohexyl) -6-methylpyrimidin-4-amine (55.49 mg,0.244mmol,1.5 eq) and 4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzoic acid (50 mg,0.163mmol,1.00 eq) in dichloromethane (5.00 mL) at room temperature were added tetramethyl chloroformyl hexafluorophosphate (182.68 mg,0.652mmol,4 eq) and N-methylimidazole (133.64 mg,1.630mmol,10 eq) under nitrogen, and the reaction solution was stirred for 1 hour. The reaction solution was cooled to room temperature, extracted with dichloromethane (3X 10 mL), the organic phases were combined, backwashed with saturated brine (1X 30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to give crude product which was purified by silica gel column chromatography, ethyl acetate/petroleum ether (5:1) to give 4-bromo-N- [2- (4, 4-difluorocyclohexyl) -6-methylpyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (48 mg, 57.10%).
MS:(ESI,m/z):516.30[M+H] + ,RT(min):1.517.
Fourth step Synthesis of N- [2- (4, 4-difluorocyclohexyl) -6-methylpyrimidin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 44):
to a solution of 4-bromo-N- [2- (4, 4-difluorocyclohexyl) -6-methylpyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (40 mg,0.077mmol,1 eq) and 2-hydroxyethanesulfonamide (11.63 mg,0.092mmol,1.2 eq) in N, N-dimethylformamide (1 mL) was added (methanesulfonic acid { bicycloethyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (7.11 mg,0.008mmol,0.1 eq), bicycloethyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine (8.28 mg,0.01 mmol) and sodium carbonate (75 mL) at room temperature, the aqueous solution was stirred and the mixture was cooled to 30mL with a saturated aqueous solution of sodium chloride solution (30 mL) and saturated with water (30 mL) was stirred to dissolve the aqueous solution, the filtrate was concentrated under reduced pressure. The crude product was purified by preparative high performance liquid phase to give N- [2- (4, 4-difluorocyclohexyl) -6-methylpyrimidin-4-yl ] -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (13.02 mg, 29.92%).
MS:(ESI,m/z):561.55[M+H] + ,RT(min):1.712.
1 H NMR:(400MHz,DMSO-d 6 )δ10.50(s,1H),10.02(s,1H),7.81(s,1H),7.51(d,J＝8.4Hz,1H),7.18(dd,J＝8.5,2.2Hz,1H),7.07(d,J＝2.3Hz,1H),5.72(s,1H),4.90(s,1H),3.76(t,J＝6.6Hz,2H),3.32-3.31(m,2H),2.87(d,J＝12.3Hz,1H),2.43(s,3H),2.29(s,2H),2.08(d,J＝10.2Hz,2H),2.01-1.74(m,8H),1.39(t,J＝6.0Hz,2H),0.22(d,J＝3.9Hz,2H),0.17(d,J＝3.8Hz,2H).
Example 14
4- ((2-hydroxyethyl) sulfonylamino) -N- [ 6-methyl-2- (pyrrolidin-1-yl) pyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 45)
First step synthesis of 6-methyl-2- (pyrrolidin-1-yl) pyrimidin-4-amine (compound 45-2):
to a solution of 2-chloro-6-methylpyrimidin-4-amine (500 mg,3.483mmol,1 eq) and pyrrolidine (297.23 mg,4.180mmol,1.2 eq) in N-methylpyrrolidone (5 mL) at room temperature under nitrogen, N-diisopropylethylamine (1.82 mL,10.449mmol,3 eq) was added. The reaction mixture was warmed to 160℃and stirred for 1 hour. The reaction mixture was cooled to room temperature and quenched with water (30 mL). The reaction mixture was extracted with ethyl acetate (3X 20 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (3X 20 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, dichloromethane/methanol (10:1) to give 6-methyl-2- (pyrrolidin-1-yl) pyrimidin-4-amine (380 mg, 61.22%).
MS:(ESI,m/z):179.15[M+H] + ,RT(min):0.471
Second step Synthesis of 4-bromo-N- [ 6-methyl-2- (pyrrolidin-1-yl) pyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 45-3):
to a solution of 6-methyl-2- (pyrrolidin-1-yl) pyrimidin-4-amine (48.74 mg,0.274mmol,1.2 eq), N, N, N ', N' -tetramethyl chloroformyl amidine hexafluorophosphate (255.75 mg,0.912mmol,4 eq) and 4-bromo-2- (spiro [2.5] oct-5-en-6-yl) benzoic acid (70 mg,0.228mmol,1.00 eq) in acetonitrile (2 mL) at room temperature was added N-methylimidazole (187.10 mg,2.280mmol,10 eq) under nitrogen. The reaction mixture was heated to 100℃and stirred for 16 hours. The reaction mixture was cooled to room temperature and quenched with water (20 mL). The reaction mixture was extracted with ethyl acetate (3X 30 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (3X 20 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (10:1) to give 4-bromo-N- [ 6-methyl-2- (pyrrolidin-1-yl) pyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (70 mg, 65.72%).
MS:(ESI,m/z):467.35[M+H] + ,RT(min):1.563
Step three synthesis of 4- ((2-hydroxyethyl) sulfonylamino) -N- [ 6-methyl-2- (pyrrolidin-1-yl) pyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 45):
to a solution of 4-bromo-N- [ 6-methyl-2- (pyrrolidin-1-yl) pyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (50 mg,0.107mmol,1 eq), 2-hydroxyethanesulfonamide (16.06 mg,0.128mmol,1.2 eq) and cesium carbonate (104.56 mg,0.321mmol,3 eq) in N, N-dimethylformamide (2 mL) were added dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine (11.44 mg,0.021mmol,0.2 eq) and (methanesulfonic acid { dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (9.56 mg,0.321mmol,3 eq) and water were then added to a solution of N, N-dimethylformamide (2 mL), and the mixture was cooled to a saturated aqueous solution (10 mm, saturated aqueous solution was cooled to a saturated solution (10 mm, and the aqueous solution was then cooled to a saturated aqueous solution was further cooled to a saturated solution of the mixture of water-saturated aqueous solution, saturated with no water, saturated aqueous solution (10 mm, saturated with no reflux; mobile phase A water (10 mmol/L ammonium bicarbonate), mobile phase B, acetonitrile; the flow rate is 60mL/min; elution gradient 30%B to 65%B in 8min; the detection wavelength is 254nm/220nm; retention time (min) 7.18. 4- ((2-hydroxyethyl) sulfonylamino) -N- [ 6-methyl-2- (pyrrolidin-1-yl) pyrimidin-4-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (25.8 mg, 46.81%) was obtained.
MS:(ESI,m/z):512.00[M+H] + ,RT(min):1.572
1 H NMR:(400MHz,DMSO-d 6 )δ9.96(s,1H),9.85(s,1H),7.51(d,J＝8.4Hz,1H),7.23–7.09(m,2H),7.05(d,J＝2.3Hz,1H),5.84–5.69(m,1H),4.95(s,1H),3.76(t,J＝6.6Hz,2H),3.42(s,4H),3.29(d,J＝6.6Hz,2H),2.26(s,5H),1.98(s,2H),1.94–1.78(m,4H),1.43(t,J＝6.0Hz,2H),0.26(d,J＝5.7Hz,4H).
Example 15
N- (2- (3-azabicyclo [3.1.0] hexane-3-yl-6-methylpyrimidin-4-yl) -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 46)
First step synthesis of 2- (3-azabicyclo [3.1.0] hex-3-yl-6-methylpyrimidin-4-amine (compound 46-2):
to a solution of 2-chloro-6-methylpyrimidin-4-amine (500 mg, 3.4813 mmol,1 eq) in N-methylpyrrolidone (5 mL,51.850mmol,14.89 eq) was added N, N-diisopropylethylamine (1350.36 mg,10.449mmol,3 eq) and 3-azabicyclo [3.1.0] hexane (434.29 mg,5.224mmol,1.5 eq) at room temperature under nitrogen. The reaction system was subjected to microwave reaction at 160℃for 2 hours. The reaction was cooled to room temperature and quenched by the addition of water (20 mL). The reaction mixture was extracted with ethyl acetate (3X 30 mL). The organic phases were combined, backwashed with saturated saline (2X 20 mL) and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel, petroleum ether/ethyl acetate (5:1) to give 2- (3-azabicyclo [3.1.0] hex-3-yl-6-methylpyrimidin-4-amine (230 mg, 34.71%).
MS:(ESI,m/z):191.10[M+H] + ,RT(min):0.608
Second step Synthesis of N- (2- (3-azabicyclo [3.1.0] hexane-3-yl-6-methylpyrimidin-4-yl) -4-bromo-2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 46-3):
To a solution of 4-bromo-2- (spiro [2.5] oct-5-en-6-ylbenzoic acid (30 mg,0.099mmol,1.00 eq), 2- (3-azabicyclo [3.1.0] hex-3-yl-6-methylpyrimidine-4-amine (27.87 mg,0.150mmol,1.5 eq) in dichloromethane (2 mL) was added N, N, N ', N' -tetramethyl chloroformamidine hexafluorophosphate (438.42 mg,1.56mmol,4 eq), N-methylimidazole (320.73 mg,3.900mmol,10 eq) was heated to 80℃and the reaction mixture was stirred for 2 hours, water (3 mL) was added to quench the reaction mixture, the organic phase was extracted with ethyl acetate (3X 30 mL), the mixture was combined with saturated brine (3X 30 mL), the residue obtained was filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure, and purified by petroleum ether chromatography (5:5-bromoimidazole (320.73 mg,3.900 eq), and the residue obtained from N-methylimidazole [ 3.27 mg, 5] was backwashed by using ethyl acetate (3.30 mL).
MS:(ESI,m/z):478.90[M+H] + ,RT(min):1.114
Step three synthesis of N- (2- (3-azabicyclo [3.1.0] hexane-3-yl-6-methylpyrimidin-4-yl) -4- ((2-hydroxyethyl) sulfonylamino) -2- (spiro [2.5] oct-5-en-6-yl benzamide (compound 46):
to a solution of N- (2- (3-azabicyclo [3.1.0] hexane-3-yl-6-methylpyrimidin-4-yl) -4-bromo-2- (spiro [2.5] oct-5-en-6-ylbenzamide (27 mg,0.056mmol,1 eq), 2-hydroxyethanesulfonamide (10.57 mg,0.084mmol,1.5 eq) in N, N-dimethylformamide (1 mL) was added dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine alkane (6.02 mg,0.01 mmol,0.2 eq), (methanesulfonic acid { dicyclohexyl (3-isopropoxy-2 ',4', -ne ', -4', 6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (5.17 mg,0.006mmol,0.1 eq), cesium carbonate (55.05 mg,0.168mmol,3 eq.) the reaction solution was warmed to 100 ℃, stirred for 1 hour, the reaction solution was cooled to room temperature, quenched with water, extracted with ethyl acetate (3X 30 mL.) the combined organic phases were backwashed with saturated brine (3X 30 mL), dried over anhydrous sodium sulfate, the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure, the crude product was purified by high performance liquid phase under the following conditions: column size XBridge Prep OBD C, 30 x 150mm,5 μm; mobile phase A water (10 mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL/min; elution gradient: 46%B to 73%B in 8min; detection wavelength: 220nm; retention time (min): 6.72. n- (2- (3-azabicyclo [3.1.0] hexane-3-yl-6-methylpyrimidin-4-yl) -4- ((2-hydroxyethyl) sulfonylamino) -2- { spiro [2.5] oct-5-en-6-yl } benzamide (6.25 mg, 21.00%),
MS:(ESI,m/z):524.00[M+H] + ,RT(min):1.633
1 H NMR:(400MHz,DMSO-d 6 )δ10.00(s,1H),9.89(s,1H),7.50(d,J＝8.3Hz,1H),7.17(dd,J＝8.5,2.3Hz,2H),7.04(d,J＝2.2Hz,1H),5.74(s,1H),4.95(s,1H),3.75(d,J＝7.3Hz,4H),3.35(m,4H),2.26(s,2H),2.24(s,3H)1.97(s,2H),1.63–1.57(m,2H),1.42(t,J＝6.0Hz,2H),0.73–0.66(m,1H),0.24(d,J＝7.6Hz,4H),0.07(q,J＝4.3Hz,1H).
Example 16
N- [6- (4, 4-Difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -4- [ (1-hydroxy-2-methylpropan-2-yl) amino ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 47)
First step Synthesis of N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -4- [ (1-hydroxy-2-methylpropan-2-yl) amino ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 47):
to a solution of 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (50 mg,0.097mmol,1 eq) in 1, 4-dioxane (3 mL) was added successively 2-amino-2-methyl-1-propanol (10.36 mg,0.116mmol,1.2 eq), (methanesulfonic acid { dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (5.18 mg,0.010mmol,0.1 eq), dicyclohexyl (3-isopropyl-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine (17.79 mg,0.019mmol,0.2 eq) and cesium carbonate (94.64 mg,0.290mmol,3.00 eq), the reaction solution was warmed to 60 ℃ and stirred for 1 hour, the reaction solution was cooled to room temperature, quenched with water (10 mL), extracted with ethyl acetate (3×10 mL), the organic phases were combined, backwashed with saturated brine (2×10 mL), dried over anhydrous sodium sulfate, filtered, the filtrate concentrated under reduced pressure and the crude product purified by high performance liquid phase under the following conditions: column gauge XSelect CSH Fluoro Phenyl μm,30mm x 150mm; mobile phase a water (0.1% formic acid), mobile phase B, acetonitrile; the flow rate is 60ml/min; elution gradient 25%B to 50%B in 8min; the detection wavelength is 254nm/220nm; retention time (min) 7.25, N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -4- [ (1-hydroxy-2-methylpropan-2-yl) amino ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (3.02 mg, 5.78%) was obtained.
MS:(ESI,m/z):525.30[M+H] + ,RT(min):2.302
1 H NMR(400MHz,CDCl 3 )δ8.74(s,1H),8.36(s,1H),7.88(d,J＝8.7Hz,1H),7.56(s,1H),6.93(d,J＝9.0Hz,1H),6.75(s,1H),6.28(s,1H),5.97(s,1H),3.92(s,2H),3.69(s,4H),2.30(s,5H),2.13(s,2H),1.98(s,4H),1.52(s,2H),1.36(s,6H),0.31(s,4H).
Example 17
N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -4- { [1- (hydroxymethyl) cyclopropyl ] amino } -2- { spiro [2.5] oct-5-en-6-yl } benzamide (compound 48)
First step Synthesis of N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -4- { [1- (hydroxymethyl) cyclopropyl ] amino } -2- { spiro [2.5] oct-5-en-6-yl } benzamide (Compound 48):
to a solution of 4-bromo-N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -2- { spiro [2.5] oct-5-en-6-yl } benzamide (50 mg,0.097mmol,1 eq) in 1, 4-dioxane (3 mL) was added (1-aminocyclopropyl) methanol (10.12 mg,0.116mmol,1.2 eq), (methanesulfonic acid { dicyclohexyl (3-isopropoxy-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine } (2 '-methylamino-1, 1' -biphenyl-2-yl) palladium (II) (5.18 mg,0.010mmol,0.1 eq) at room temperature, dicyclohexyl (3-isopropyl-2 ',4',6 '-triisopropyl- [1,1' -biphenyl ] -2-yl) phosphine (17.79 mg,0.019mmol,0.2 eq) and cesium carbonate (22.11 mg,0.29 mmol,3 eq), the reaction mixture was warmed to 60 ℃ and stirred for 1 hour, the reaction mixture was cooled to room temperature, quenched with water (10 mL), extracted with ethyl acetate (3×10 mL), the organic phases were combined, backwashed with saturated brine (1×10 mL), dried over anhydrous sodium sulfate, filtered, the filtrate concentrated under reduced pressure and the crude product purified by high performance liquid phase under the following conditions: the chromatographic column specification is XBridge BEH Shield RP, 30mm is 150mm,5 μm; mobile phase A water (10 mmol/L ammonium bicarbonate), mobile phase B, acetonitrile; the flow rate is 60ml/min; elution gradient 53%B to 72%B in 10min; the detection wavelength is 254nm/220nm; retention time (min) 8.47. N- [6- (4, 4-difluoropiperidin-1-yl) -4-methylpyridin-2-yl ] -4- { [1- (hydroxymethyl) cyclopropyl ] amino } -2- { spiro [2.5] oct-5-en-6-yl } benzamide (2.30 mg, 4.20%) was obtained.
MS:(ESI,m/z):523.50[M+H] + ,RT(min):3.147
1 H NMR(400MHz,CDCl 3 )δ8.83(s,1H),7.86(d,J＝8.5Hz,1H),7.61(s,1H),6.75(dd,J＝8.6,2.4Hz,1H),6.47(d,J＝2.5Hz,1H),6.27(s,1H),5.94(s,1H),4.68(s,1H),3.72(s,4H),3.68(s,2H),2.31(s,5H),2.14(s,2H),2.02(d,J＝14.5Hz,4H),1.25(s,2H),1.20(s,1H),0.89(s,4H),0.51–0.15(m,4H).
Biological evaluation
Test example 1
Test name: imaging-based Nuclear Count Analysis (NCA) in OVCAR-3 cells
Day 0 compound dilution and treatment
a) The final test concentrations of AM-5308 were: 10000. 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, 0.5nM.
b) Test compounds, final test concentrations were: 10000. 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, 0.5nM.
c) Cells were incubated at 37℃with 5% CO 2 Is cultured in an incubator for 4 days.
d) DMSO concentration was 0.1%.
Day 1 cells were seeded into 384 well cell culture plates
a) When the cell confluence reaches 80% -90%, the cells are treated.
b) Cells were resuspended in medium and then counted and diluted at the desired density.
c) 30. Mu.L/well of a cell suspension containing the appropriate cells was added to 384 well plates: 600 cells/well.
Day 4 assay
a) mu.L of 8% fixative (final concentration 4%) was added and the plates incubated for 30 min at room temperature.
b) Centrifugal plate, 1000rpm,30s.
c) The cells were washed twice with 60. Mu.l/well PBS.
d) After fixation, cells were permeabilized and stained in 60. Mu.L wash buffer (1% BSA, 0.2% Triton X-100, 1 XPBS) containing 2. Mu.g/mL Hoechst 33342DNA dye.
e) The plates were sealed and incubated at room temperature for 1 hour in the dark.
f) Wash 3 times with PBS.
g) Add 50. Mu.LPBS/well and scan the plate using HCS.
h) Data acquisition and detection
Data analysis
Inhibition (%) = 100- (compound well read-low read control well read)/(high read control well read-low read control well read) ×100
High read control wells: adding 30nL DMSO to the cells; low read control wells: 10. Mu.M AM-5308 wells.
Calculation of IC using GraphPad Prism 8 software 50 (nM) and plotting the effect of the compounds versus dose curve.
TABLE 1 cell Activity data for the compounds of the present application
Test example 2
Test name: ADP-Glo TM Kinase assay
The operation steps are as follows:
1) 1 Xthe reaction buffer was prepared.
2) 100nL of diluted stock solution of compound was transferred to each well of the reaction plate using Echo 655. The final concentration of DMSO was 1%.
3) The reaction plate was sealed with a sealing plate membrane and centrifuged at 1000g for 1 min.
4) A2 Xenzyme solution was prepared with 1 Xreaction buffer.
5) mu.L of 2 Xenzyme solution was added to each well of the reaction plate. 1000g of the plate is sealed by a sealing plate membrane and centrifuged for 1 min, and the plate is placed for 15 min at room temperature.
6) A2X ATP solution was prepared with 1X reaction buffer.
7) To the reaction plate, 5. Mu.L of a 2X ATP solution was added, and the mixture was centrifuged at 1000g for 1 minute to start the reaction.
8) The reaction was carried out at room temperature for 60 minutes.
9) Add 10. Mu.L ADP Glo reagent. Centrifuge 1000g for 1 min and incubate at room temperature for 60 min.
10 20. Mu.L of kinase assay reagent was added. Centrifuge 1000g for 1 min and incubate at room temperature for 60 min.
11 1000g for 1 minute.
12 At Envision 2104).
Data analysis:
percent inhibition was calculated as follows:
% inhibition = 100- (Signal) cmpd -Signal Ave_PC )/(Signal Ave_VC -Signal Ave_PC )×100
Signal cmpd Average value of test compounds on the reaction plate.
Signal Ave_PC Mean value of positive control on the reaction plate.
Signal Ave_VC Mean value of negative control on the reaction plate.
Computing IC 50 Fitting a compound dose-response curve:
IC of the compound was obtained using a nonlinear fitting equation using GraphPad 8.0 50 。
3) Quality control
Z factor>0.5；S/B>2。
TABLE 2 enzymatic Activity data for Compounds of the present application
The embodiments of the technical solution of the present invention have been described above by way of example. It should be understood that the protection scope of the present invention is not limited to the above embodiments. Any modification, equivalent replacement, improvement, etc. made by those skilled in the art within the spirit and principles of the present invention should be included in the scope of the claims of the present application.

Claims (10)

1. A compound of formula (I), racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof:
Wherein,
X 1 、X 2 、X 3 identical or different, independently of one another, from N or CR 0 ；R 0 Selected from H, halogen, cyano, C 1-12 Alkyl, halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halo C 1-12 Alkoxy, cyano C 1-12 An alkoxy group;
a is selected from unsubstituted or optionally substituted with one, two or more R a Substituted with the following groups: c (C) 1-12 Alkyl, halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halo C 1-12 Alkoxy, cyano C 1-12 Alkoxy, C 3-12 Cycloalkyl, C 3-12 Cycloalkyl oxy, C 3-12 Cycloalkylthio, 3-14 membered heterocyclyl; each R a The same or different, independently of one another, are selected from halogen, cyano, C 1-12 Alkyl, halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, halo C 1-12 Alkoxy, cyano C 1-12 An alkoxy group;
Y 1 、Y 2 、Y 3 the same or different, independently of one another, from N or CH;
m is selected from unsubstituted or optionally substituted with one, two or more R b Substituted with the following groups: c (C) 3-12 Cycloalkyl, C 3-12 Cycloalkenyl, 3-14 membered heterocyclyl; each R b The same or different, independently of one another, are selected from halogen, cyano, C 1-12 Alkyl, halogenated C 1-12 Alkyl, cyano C 1-12 Alkyl, C 1-12 Alkoxy, cyano C 1-12 An alkoxy group;
e is selected from unsubstituted or optionally substituted with one, two or more R c Substituted with the following groups: -NH-S (=o) 2 -R c1 、-S(＝O) 2 -NH-R c2 、-S(＝O)(＝NH)-R c3 、-N(R c4 )(R c5 ) 3-14 membered heterocyclyl; each R c The same or different, independently of one another, from OH, halogen, cyano, C 1-12 Alkyl, C 1-12 Alkoxy, halo C 1-12 Alkyl, halogenated C 1-12 Alkoxy, cyano C 1-12 Alkyl, cyano C 1-12 Alkoxy, -N (R) c6 )(R c7 )；
R c1 、R c2 、R c3 、R c4 、R c5 、R c6 、R c7 Identical or different, independently of one another, from H, C 1-12 Alkyl, hydroxy C 1-12 Alkyl, halogenated C 1-12 Alkyl, halogenated C 1-12 Alkoxy, cyano C 1-12 Alkyl, cyano C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclyl, C 1-12 alkoxy-C 1-12 Alkyl, hydroxy C 1-12 alkyl-C 3-12 Cycloalkyl groups.
2. A compound according to claim 1, wherein X 2 、X 3 Not both are N;
preferably, when X 1 And X 2 Is N or CR 0 When X is 3 Is CR (CR) 0 ，R 0 Selected from H, halogen, CN, C 1-6 Alkyl groupThe method comprises the steps of carrying out a first treatment on the surface of the When X is 1 Is N or CH, X 2 When CH is, X 3 Is N or CR 0 ，R 0 Selected from H, halogen, CN, C 1-6 An alkyl group; when X is 1 Is N or CR 0 ，X 2 When N is N, X 3 Is N or CR 0 ；R 0 Selected from H, halogen, CN, C 1-6 An alkyl group;
preferably, when X 1 And X 2 When N is N, X 3 Is CR (CR) 0 ，R 0 Selected from H, C 1-6 An alkyl group; when X is 1 Is N or CH, X 2 When CH is, X 3 Is N or CR 0 ，R 0 Selected from H, C 1-6 An alkyl group;
preferably, A is selected from C 1-6 Alkyl, C 3-8 Cycloalkyl, C 3-8 Cycloalkyloxy, 3-8 membered heterocyclyl, halogenated C 3-8 Cycloalkyl;
preferably, A is selected from cyclopropyl, isopropyl,
Preferably, when Y 1 When CH is, Y 2 Is CH, Y 3 Is N or CH; when Y is 1 When CH is, Y 2 Is N, Y 3 Is N or CH; when Y is 1 When N is N, Y 2 Is CH, Y 3 Is N or CH.
3. A compound according to claim 1 or 2, wherein M is selected from C 3-12 Cycloalkyl, C 3-12 Cycloalkenyl, 3-12 membered heterocyclyl. Preferably, M is selected from C 3-12 Condensed ring cycloalkyl, C 3-12 Condensed ring cycloalkenyl, 3-12 membered condensed ring heterocyclyl, C 3-12 Spiro cycloalkyl, C 3-12 Spirocyclic cycloalkenyl, 3-12 membered spiroheterocyclic group, C 3-12 Bridged cycloalkyl, C 3-12 Bridged cycloalkenyl or 3-12 membered bridged heterocyclyl;
preferably, M is selected from
4. A compound according to any one of claims 1-3, characterized in that E is selected from-NH-S (=o) 2 -R c1 、-S(＝O) 2 -NH-R c2 、-S(＝O)(＝NH)-R c3 、-N(R c4 )(R c5 )、C 1-6 Alkyl substituted 3-6 membered heterocyclyl;
R c1 、R c2 、R c3 、R c4 、R c5 the same or different, are independently selected from H, hydroxy C 1-6 Alkyl, halogenated C 1-6 Alkyl, cyano C 1-6 Alkyl, C 1-6 alkyl-NH-C 1-6 Alkyl-, (C) 1-6 Alkyl group 2 N-C 1-6 Alkyl-, C 3-6 cycloalkyl-NH-C 1-6 Alkyl-, C 1-6 alkoxy-C 1-6 Alkyl-, C 3-6 Cycloalkyl, 3-6 membered heterocyclyl, hydroxy-C 3-6 Cycloalkyl, hydroxy C 1-6 alkyl-C 3-6 Cycloalkyl;
preferably E is selected from
5. A compound according to any one of claims 1 to 4, wherein the compound is selected from the following structures:
therein, A, E、M、X 2 、X 3 、Y 1 、Y 2 、Y 3 Having the definition of any one of claims 1 to 4.
6. The compound according to any one of claims 1 to 5, wherein the compound has a structure represented by formula (II), preferably a structure represented by formula (III), more preferably a structure represented by formula (IV):
therein, A, E, X 1 、X 2 、X 3 、R c1 Having the definition of any one of claims 1 to 5;represents a carbon-carbon single bond or a carbon-carbon double bond.
7. A compound according to any one of claims 1 to 6, wherein the compound is selected from the following structures:
8. a process for the preparation of a compound as claimed in any one of claims 1 to 7 comprising the steps of:
(1) Reacting the compound a with the compound b to obtain a compound c;
(2) Reacting the compound c with E-H to obtain a compound shown in a formula (I);
therein, A, E, M, X 1 、X 2 、X 3 、Y 1 、Y 2 、Y 3 Having the definition of any one of claims 1 to 7 independently of one another; x is selected from halogen, such as Cl, br, I.
9. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound of any one of claims 1-7, racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof.
10. Use of at least one of the compounds of any one of claims 1-7, racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof for the manufacture of a medicament;
Preferably, the use is in the manufacture of a medicament for the treatment of a KIF18A mediated disorder and/or disease, such as in the manufacture of a KIF18A inhibitor medicament;
preferably, the disease is cancer, including bowel cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer or ovarian cancer.
CN202310972422.4A 2022-08-05 2023-08-03 KIF18A inhibitor compound, pharmaceutical composition, preparation method and application thereof Pending CN117510463A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116789637A (en) * 2022-05-13 2023-09-22 上海湃隆生物科技有限公司 Kinesin KIF18A inhibitor and application thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116789637A (en) * 2022-05-13 2023-09-22 上海湃隆生物科技有限公司 Kinesin KIF18A inhibitor and application thereof

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